Tobacco

: Production, Chemistry and

title:
Technology
author: Davis, D. Layten
publisher: Blackwell Publishing Ltd.
isbn10 | asin:
print isbn13: 9780632047918
ebook isbn13: 9780632062430
language: English
subject Tobacco, Tobacco industry.
publication date: 1999
lcc: SB273.T67 1999eb
ddc: 679/.7
subject: Tobacco, Tobacco industry.
 Page iii

Tobacco
Production, Chemistry and Technology
Edited by
D. Layten Davis
and
Mark T. Nielsen

 Page iv
© 1999 by CORESTA
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Tobacco: production, chemistry, and technology / edited by
D. Layten Davis and Mark T. Nielsen.
p. cm.
Includes bibliographical references and index.
ISBN 0-632-04791-7
1. Tobacco. 2. Tobacco industry. I. Davis, D. Layten
(Daniel Layten,), 1938- . II. Nielsen, Mark T.
SB273 .T67 1999
679'.--dc21 98-51589
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 Page v

Contents
Preface vii
List of Contributors ix
Acknowledgements xi
1 1
Seed to Smoke
T.C. Tso
2 32
Breeding and Genetics
P.D. Legg and B.W. Smeeton
3 49
Biotechnology: Uses and Applications in Tobacco
Improvement
J.D. Brandle and D. Bai
4 66
Agronomy and Physiology
4A Tobacco Seed 66
T. W. Hutchens
4B Seedling Production 70
W.D. Smith
4C Field Practices 76
K.C. Flower
5 104
Production Practices
 5A Flue-Cured Tobacco 104
G.F. Peedin
5B Light Air-Cured Tobacco 143
G.K. Palmer and R.C. Pearce
5C Oriental Tobacco 154
S.N. Gilchrist
5D Dark Fire-Cured Tobacco 164
Robert D. Miller and Donald J. Fowlkes
6 183
Major Tobacco Diseases
6A Fungal and Bacterial Diseases 183
P.B. Shoemaker and H.D. Shew
6B Virus Diseases 198
D. Blancard, R. Delon, B.W. Blair and T. Glover
6C Nematode Pests of Tobacco 216
J.A. Shepherd
7 228
Tobacco Insect Pests
7A Insects and Their Management in Tobacco 228
Production
B.W. Blair
7B Stored Tobacco: Insects and Their Control 241
E.D. Massey
7C Pesticide Regulations and Their Impact on Crop 250
Protection Strategies (Minimization of Pesticide
Residues)
L. Mueller and M.R. Ward
8 265
Leaf Chemistry
8A Basic Chemical Constituents of Tobacco Leaf 265
and Differences Among Tobacco Types
J.C. Leffingwell
8B Alkaloid Biosynthesis 285
L.P. Bush
8C Leaf Surface Chemistry 292
G. Wagner
8D Relationship between Leaf Chemistry and 304
Organoleptic Properties of Tobacco Smoke
W.W. Weeks
9 313
Physical Properties of Leaf Tobacco
Y. Nakanishi
10 320
Marketing, Processing and Storage
10A Tobacco Marketing Systems 320
John S. Campbell
10B Green Leaf Threshing and Redrying Tobacco 330
Bill Ward
10C Tobacco Storage 338
L. Ryan
11 346
Cigarette Manufacture
11A Tobacco Blending 346
Phil Fisher
 11B Cigarette Design and Materials 353
Alan Norman
11C Cigarette Production and Quality Assurance 388
J.L. McKenzie and Chris Crawley
12 398
Smoke Chemistry
Richard R. Baker
13 440
Cigars and Cigarillos
Adeler Frederik Wehlburg
14 452
Smokeless Tobacco
Inger Wahlberg and Tommy Ringberger
Index 461

 Page vii

Preface
During the twentieth century tobacco has become one of the most
economically important agricultural crops in the international
marketplace. Not only do farmers in over 100 countries depend upon
tobacco as a major source of cash income, but an entire industry, from
a diverse manufacturing sector to distribution and retail outlets, has
grown to be a major economic force in many industrial and
developing countries. Along with the growth in the tobacco industry,
local and national governments in many countries have reaped added
benefits through the collection of tax revenues. The growth in the
tobacco industry has been supported by numerous scientific and
technical advances in the last century. Indeed, one could speculate that
without these achievements the tobacco industry would not have
reached the level of global importance it has today.
Despite the rather remarkable advances for tobacco as an agricultural
industry, it would seem that few other industries have faced as many
challenges and changes in recent years. The globalization of the
tobacco industry, new legislation and efforts by external forces have
greatly altered the environment in which the tobacco industry
operates. While many in the tobacco industry have continued to strive
to produce a high quality tobacco leaf for superior consumer products,
the new, dynamic environment has made it difficult to keep that focus.
Indeed, many individuals throughout the tobacco industry are
focusing on new issues to more effectively meet the needs of this
ever-changing environment. These challenges should be considered
positive, as it will be through effectively meeting these challenges that
the tobacco industry will provide for the next hundred years.
The industry has experienced cyclic supplies of quality leaf for a long
time some years have a high volume of leaf, others a rather low
volume. Naturally, as supply cycles world prices fluctuate
significantly. The tobacco farmer is at the mercy of these production
cycles and the industry has responded with attempts to minimize this
cyclic production to a more constant supply flow. The impact of
external issues such as the anti-smoking efforts makes it difficult to
predict future demand, but nevertheless, the production of a
stablesupply of high quality leaf tobacco will remain important to
growers and others throughout the industry.
Many factors have contributed to fluctuations in the global supply of
tobacco leaf. The common trend seems to be a short supply, followed
by price increases, followed by over-production. This trend can be
exacerbated by a number of factors. In some countries, policies
designed to support production will contribute to excess production
and an over-supply of tobacco leaf. For example, regulations in the
USA that limit quota reductions may actually enhance the magnitude
of the over-supply part of the cycle on a worldwide basis. Also, the
supply side of the equation is affected by changes in government
regulations that affect tobacco and by differences among countries in
regulations that affect the movement of tobacco throughout the world
markets. On the other hand, tobacco diseases and insects, unfavorable
weather, increased production expenses, the need to conserve soils
and low prices will drive down tobacco production.
On a worldwide basis, the demand for tobacco continues to increase,
although not at the pace set a few years ago. Cigarette consumption in
many developed countries, the USA in particular, has tended to
decline over the past decade, but this downward trend appears to be
leveling out. Further government regulations, including restrictions on
advertising, could have a further negative effect on the demand for
tobacco products. To achieve some stability in the global supply of
tobacco and to ensure that growers will continue to produce a high
quality leaf, scientific advancements must continue, and growers and
others in the industry must adapt and plan to meet the needs of the
consumer.
In recent years, this is perhaps best exemplified by the price-conscious
consumer driving up the demand for 'value brand' products in some
countries. Such 'house' brands require careful monitoring and taste test
screening of less expensive tobaccos. The industry found that
although consumers request a lower-priced product, they are not and
should not be willing to accept a taste trade-off.
Shifts in global demands for new or existing products, primarily
blended cigarettes and cigars, are

 Page viii
changing tobacco production requirements. These changes may
necessitate that the tobacco industry place special emphasis on
scientific advancements in variety development, agronomic practices,
pest control and leaf curing to meet the needs of consumers in many
different countries. This would make tobacco product manufacture a
more segmented industry, but it would permit the industry to be more
responsive to each market.
While consumer preferences drive raw product requirements, it is the
purchaser of the leaf that must implement many of these changes.
Leaf quality and price are often deciding factors in the implementation
effort. As we move to the future of tobacco products, as well as other
consumer products, we will see an increasing use of biotechnology to
enhance a product's usability and value. Because of certain unique
characteristics of the tobacco plant, it has been widely used in genetic
studies for over 75 years. Rapid advancements in knowledge of
genetics and technological creativity have provided remarkable tools
to genetically improve the tobacco plant. These improvements could
be targeted towards fine-tuning the plant to achieve certain
characteristics including improved agronomic performance and pest
resistance.
Many of the advancements in tobacco science have arisen from
research conducted at public institutions including universities and
government agencies. A reduction in public support for future tobacco
research has already occurred in some countries. This will
undoubtedly make it more difficult to address the research needs of
growers and manufacturers, as well as hamper the development of the
next generation of tobacco scientists. Broad-based support for
research will have to come from within the industry, and
consequently, research programs must be carefully evaluated for their
cost effectiveness, value to the industry and the ease of supplying the
research results at the farm level.
This CORESTA monograph will discuss the interrelationships among
the growth of the tobacco plant, the harvested leaves, their curing,
processing and manufacturing, and the properties of the final product.
Through each step of the process from tobacco seed germination to
smoke yield the goal of the entire process is consumer satisfaction. A
discussion of the breeding and genetics of the tobacco plant flows into
a chapter about the new frontier of biotechnology. Tobacco is an ideal
recipient for the introduction and expression of foreign genes for use
in plant enhancement or disease resistance. Biotechnology will
inevitably have an impact on future tobacco production and
utilization. The combined study of the tobacco plant's physiology and
the improvement of agronomic practices has enabled and will
continue to enable us to enhance the leaf yield. This monograph will
outline the general management practices for each of the major
tobacco types flue-cured, light air-cured, Oriental, cigar and fire-
cured. The economic losses, management, control, effects on
tobacco's chemical composition and leaf usability will be studied in a
chapter outlining major tobacco diseases. The minimization of
pesticide residues is a worldwide issue and is featured in a discussion
of tobacco insect management from production to storage of tobacco
products. The basic chemical constituents of the tobacco leaf and the
differences among tobacco types are presented in a chapter on leaf
chemistry, followed by a discussion of tobacco's physical properties in
relationship to manufacturing needs and properties. Tobacco's
marketing systems, threshing and redrying, aging, fermentation and
storage procedures blend into a monograph section on cigarette
design. This monograph will closely examine current practices and
new developments in the area of cigarette manufacture and the study
of smoke chemistry. Lastly, the topics related to cigars, cigarillos and
smokeless tobacco products will be explored. This monograph is
intended to offer a broad view of current tobacco knowledge /
practices and features sections relating to the future of tobacco.
This monograph highlights the current state of knowledge in the areas
just described. To accomplish this, we asked our contributing authors
to focus their attention on the up-to-date scientific literature as it
relates to each chapter topic. It was not our intention to compile a
complete compendium of the literature on tobacco science and
technology, nor did we intend to provide a thorough history of
tobacco. We do hope, however, that the reader will find that this
monograph provides a rich source of information about the art,
science and technology of tobacco.
D. LAYTEN DAVIS
R. J. REYNOLDS TOBACCO COMPANY
WINSTON-SALEM, NORTH CAROLINA, USA

MARK T. NIELSEN
UNIVERSITY OF KENTUCKY
LEXINGTON, KENTUCKY, USA

 Page ix

List of Contributors
D. Bai, Imperial Tobacco Ltd, Montreal, Quebec, Canada
(3 Biotechnology: Uses and Applications in Tobacco Improvement)
Richard R. Baker, British American Tobacco, Southampton, United
Kingdom
(12 Smoke Chemistry)
B. W. Blair, Tobacco Research Board, Harare, Zimbabwe
(6B Virus Diseases and 7A Insects and their Management in Tobacco
Production)
D. Blancard, Institut du Tabac, Seita, Bergerac, France
(6B Virus Diseases)
J. D. Brandle, Agriculture and Agri-Food, Delhi, Ontario, Canada
(3 Biotechnology: Uses and Applications in Tobacco Improvement)
L. P. Bush, University of Kentucky, Lexington, Kentucky, USA
(8B Alkaloid Biosynthesis)
John S. Campbell, John S. Campbell Ltd, Wilson, North Carolina,
USA
(10A Tobacco Marketing Systems)
Chris Crawley, Fidus Instrument Corporation, Richmond, Virginia,
USA
(11C Cigarette Production and Quality Assurance)
L. Davis, R.J. Reynolds Tobacco Company, WinstonSalem, North
Carolina, USA
(Preface)
R. Delon, Institut du Tabac, Seita, Bergerac, France
(6B Virus Diseases)
Phil Fisher, Tobacco Consultant, Louisville, Kentucky, USA
(11A Tobacco Blending)
K. C. Flower, Tobacco Research Board, Harare, Zimbabwe
(4C Field Practices)
Donald J. Fowlkes, University of Tennessee, Knoxville, Tennessee,
USA
(5D Dark Fire-cured Tobacco)
S. N. Gilchrist, R. J. Reynolds Tobacco Company, WinstonSalem,
North Carolina, USA
(5C Oriental Tobacco)
T. Glover, Tobacco Research Board, Harare, Zimbabwe
(6B Virus Diseases)
T. W. Hutchens, F. W. Rickard Seeds, Inc., Lexington, Kentucky, USA
(4A Tobacco Seed)
J. C. Leffingwell, Leffingwell and Associates, Canton, Georgia, USA
(8A Basic Chemical Constituents of Tobacco Leaf and Differences
among Tobacco Types)
P. D. Legg, University of Kentucky, Lexington, Kentucky, USA
(2 Breeding and Genetics)
E. D. Massey, British American Tobacco, R&D, Southampton, UK
(7B Stored Tobacco: Insects and their Control)
J. L. McKenzie, McKenzie and Rains, WinstonSalem, North Carolina,
USA
(11C Cigarette Production and Quality Assurance)
Robert D. Miller, University of Tennessee, Knoxville, Tennessee,
USA
(5D Dark Fire-cured Tobacco)
L. Mueller, R. J. Reynolds Tobacco GmbH, Cologne, Germany
(7C Pesticide Regulations and their Impact on Crop Protection
Strategies (Minimization of Pesticide Residues))
Y. Nakanishi, Japan Tobacco Inc., Yokohama, Japan
(9 Physical Properties of Leaf Tobacco)
M. Nielsen, University of Kentucky, Lexington, Kentucky, USA
(Preface)
Alan Norman, R. J. Reynolds Tobacco Company, WinstonSalem,
North Carolina, USA
(11B Cigarette Design and Materials)
G. K. Palmer, University of Kentucky, Lexington, Kentucky, USA
(5B Light Air-cured Tobacco)
R. C. Pearce, University of Kentucky, Lexington, Kentucky, USA
(5B Light Air-cured Tobacco)
G. F. Peedin, North Carolina State University, Raleigh, North
Carolina, USA
(5A Flue-cured Tobacco)

 Page x
Tommy Ringberger, Swedish Match Sverige AB, Stockholm, Sweden
(14 Smokeless Tobacco)
L. Ryan, Philip Morris Europe, Neuchâtel, Switzerland
(10C Tobacco Storage)
J. A. Shepherd, Tobacco Research Board, Harare, Zimbabwe
(6C Nematode Pests of Tobacco)
H. D. Shew, North Carolina State University, Raleigh, North Carolina,
USA
(6A Fungal and Bacterial Diseases)
P. B. Shoemaker, North Carolina State University, Raleigh, North
Carolina, USA
(6A Fungal and Bacterial Diseases)
B. W. Smeeton, R. J. Reynolds Tobacco Company, Winston-Salem,
North Carolina, USA
(2 Breeding and Genetics)
W. D. Smith, North Carolina State University, Raleigh, North
Carolina, USA
(4B Seedling Production)
T. C. Tso, Ideals Inc, Beltsville, Maryland, USA
(1 Seed to Smoke)
G. Wagner, University of Kentucky, Lexington, Kentucky, USA
(8C Leaf Surface Chemistry)
Inger Wahlberg, Swedish Match Sverige AB, Stockholm, Sweden
(14 Smokeless Tobacco)
M. R. Ward, Advanced Technologies (Cambridge) Limited,
Cambridge, United Kingdom
(7C Pesticide Regulations and their Impact on Crop Protection
Strategies (Minimization of Pesticide Residues))
Bill Ward, Export Leaf Tobacco Company, Wilson, North Carolina,
USA
(10B Green Leaf Threshing and Redrying Tobacco)
W. W. Weeks, North Carolina State University, Raleigh, North
Carolina, USA
(8D Relationship between Leaf Chemistry and Organoleptic
Properties of Tobacco Smoke)
Adeler Frederik Wehlburg, ASP Enterprises, Inc., Guayaquil, Ecuador
(13 Cigars and Cigarillos)

 Page xi

Acknowledgements
The editors express appreciation to CORESTA and to CORESTA's
Scientific Commission for providing the opportunity to publish this
Tobacco Monograph. The assistance of Francois Jacob is especially
noteworthy.
The contributions of all chapter authors and H. Burton, who
coordinated the leaf chemistry chapter, are greatly appreciated.
H. Papenfus and E.A. Wernsman contributed significantly to the
original idea for this monograph. In addition, H. Papenfus was
involved with the selection of the chapter topics.
The excellent assistance of Ann Niten, who formated and incorporated
numerous revisions into the monograph, and the dedicated efforts of
Veda Davis, who proofed the entire monograph and revisions, are
recognized.
We also thank Patty Turner for her assistance in organizing and
preparing the monograph. Special thanks are extended to H. Chung
and the RJRT Library staff for their work in locating the correct
reference citations.
Further, we acknowledge the contributions of Sue Moore, Blackwell
Science Senior Editor, who was very helpful and encouraging during
the preparation and publication phases of this monograph.
Appreciation is expressed to the persons who reviewed the
manuscripts. They include R. Black, L.P. Bush, J. Chappell, D.
Fleming, B. Fortnum, C. Green, D. Hill, A. Johnson, B. Kennedy, C.
Lily, W. Lloyd, R.C. Long, R. Manning, R. Monk, Jr, W. Nesmith, R.
Pearce, H. Papenfus, T. Parish, T.J. Porter and A. Rodgman.
Finally, we acknowledge the importance to the authors of this
monograph of the previous publication Tobacco by B.C. Akehurst
(Longman Group Limited, Essex, UK). Numerous sections utilize
information as evidenced by the citations. That book has served as a
major reference for this industry for over three decades.

 Page 1

Chapter 1
Seed to Smoke
T.C. Tso
Ideals, Inc.
Beltsville, Maryland, USA
Introduction
a
Background
From seed to smoke the culture, manufacture and use of tobacco
involves a continuous chain of events. Each step in those events
represents a single link of the long chain, and each link is of equal
significance. Weakening of any single link produces a product of
inferior quality; breaking the chain or eliminating any single link may
produce a nonusable product (Tso, 1990).
Tobacco is similar to most agricultural products in that it begins with a
single seed which is a carrier of genetic information. Environmental
elements provide the proper conditions needed for the full expression
of such genetic information. Human intervention through cultural
practices, for example, influences the degree or fullness of gene
expression, such as development of plant and leaf characteristics in
the field. Postharvest manipulation charts the course of physiological
and biochemical changes toward desired quality and thus usability.
Tobacco differs from other crops in that it is used mostly for
combustion. Variables of botanical, physical and chemical
characteristics of leaf tobacco determine degrees of combustibility,
smoke composition, taste and aroma and, thus, product acceptability.
From seed to smoke involves a long, stepwise process, which is a
science as well as an art. Tobacco (Nicotiana tabacum L.) is one of the
most, if not the most, studied species today in plant and biological
science, physics and chemistry, and in bioengineering and technology.
For example, in one month alone (May 1996) there were 231
scientific reports involving tobacco. All in all, much is known about
tobacco, but much has yet to be learned and much has yet to be fully
understood and appreciated.
b
Classification of Tobacco
Most of the commercial tobaccos produced in the world are Nicotiana
tobacum. The only other species used on a limited commercial scale is
N. rustica. In addition to the botanical classification of the species,
tobacco is classified on the basis of major types and general uses.
Because the properties of tobacco and, therefore, its usability vary
markedly with variety, locality, system of production and curing
method, standardization of the commercial product is essential for
growers and users (i.e. manufacturers). It is based primarily on curing
method (air-, sun-, fire-and flue-curing), locality of production
(growth) and the way in which the leaf is to be used (cigarette, cigar,
pipe, etc.). Further classification is then according to position on the
stalk from which the leaves have originated and factors such as their
color, quality and ripeness at harvest.
Flue-cured tobaccos, for example, are broadly separated into
categories by country such as the USA, Brazil, Zimbabwe or China.
Within each such broad geographical division specific growing
regions may be identified by virtue of distinctive characteristics, e.g.
Old Belt, Middle Belt, Eastern North Carolina, South CarolinaNorth
Carolina Border Belt and GeorgiaFlorida in the USA and Highveld
and Lowveld in Zimbabwe. The product may be used mainly as filler
or flavor components of cigarette and pipe blends.
The main types of tobacco and their production statistics based on
forecast values for 1996 are shown in Table 1.1.
c
Quality and Usability
Quality, in the common sense of the word, means the desired
characteristics or status of a certain product or format at a given time,
location and individual lot. Quality represents a balance of essential
properties which meets the preference of a group of consumers at

 Page 2
Table 1.1 The main types of tobacco, area under
production and production volume (forecast
values, 1996).
Tobacco Area Volume (metric Volume
(Type) (ha) tons) (%)
Flue-cured 2 462 4 533 995 63.3
967
Dark air/sun- 818 981 172 13.7
cured 312
Burley 499 844 750 11.8
455
Oriental 457 570 574 8.0
506
Light air-cured 64 779 89 316 1.2
Dark air-cured, 96 439 87 148 1.2
cigar
Dark fired- 56 798 58 608 0.8
cured
Total 4 456 7165 563 100
256
Source: USDA (1996).

a particular time and location. In other words, the concept of quality

or conditions is relative, which may change with location, time,
individual lot or personal assessment.
In tobacco, quality factors relate to visible, sensory, physical and
chemical properties of the leaf and extend to its combustion product,
the smoke. In the final analysis, quality relates to acceptability of the
balance of these factors.
Usability is a measure of how suitable leaf is for a specific blend and
manufacturing process. In addition to the various quality attributes,
price comes into the definition. The same grade from a particular
growing area may be very usable for one product or brand because of
its smoking and physical characteristics and cost, but may rate poorly
for another. Therefore, it may be in greater demand than, say, a grade
that is more attractive in terms of its classification.
Genetic Makeup
a
Botanical Characteristics
There are significant botanical variations within the Nicotinia genus.
Tobacco germplasms have been collected over several decades and
deposited in a United States Department of Agriculture (USDA)
inventory. This collection numbers over 1500 entries which differ in
botanical characteristics such as plant habit, height and the
morphology of its leaves (leaf shape, thickness and size, form of leaf
tip, petiolate or sessile, leaf angle to stem, leaf asymmetry, etc.). This
USDA collection of tobacco introductions (TI) includes species,
intraspecific and interspecific hybrids, monosomic, plus established
varieties, mutant lines and breeding lines. Each one has its unique
genetic make-up and thus botanical characteristics (Tso, 1972; Tso,
1990; Tso, 1996).
Tobacco originated in South and Central America, probably from the
mid to low altitude forest margins by virtue of its small seed size, light
sensitivity to germination, low light saturation for photosynthesis,
relatively large leaf area and susceptibility to frost. It is naturally a
perennial, but is farmed as an annual crop.
Most commercial tobaccos belong to the species Nicotiana tabacum
L., an allotetraploid that resulted from natural hybridization of the two
wild species N. sylvestris Speg and Comes and N. tomentosiformis
Goodsp (Wernsman & Matzinger, 1980). in the past some researchers
have proposed N. octophora. Current commercial varieties of tobacco
probably owe their survival to man based on the fact they have
apparently never been found in the wild state.
Nicotiana tabacum L. produces its leaves from a single, erect stem
with a terminal inflorescence. Although considerable variation exists,
as shown by the collection of 1500 or so tobacco germplasms held by
the USDA, the leaves are typically ovate and oblonglanceolate. They
are formed spirally and show heteroblastic development. In flue-cured
varieties grown as commercial crops, leaf area ranges from about6
dm2 at lower nodes to 18 dm2 in the middle stalk and 12 dm2 near the
apex. Cigar varieties have slightly smaller leaves, whereas Oriental
tobaccos are cultured to produce particularly small leaves.
Leaf and stem surfaces are covered to a greater orlesser extent by
trichomes that produce exudates con-taining important precursors to
the compounds whichcontribute to the distinctive aroma and flavor of
thecured product. Also, the trichomes and their exudateshave a key
role in insect susceptibility and resistance.
The varieties developed in the USA produce a largethick stalk, with a
woody lower portion. In most ofthese varieties the leaf does not have
a bare petiole. Inthe leaf axils of the stalk, there are axillary buds
thatordinarily remain dormant during the growth period,but which, if,
the terminal bud is removed, readilydevelop into large branches
commonly known assuckers. During the maturation phase, a few
suckersmay develop even when the terminal bud (inflorescence)
remains intact. Also, certain weather conditionsmay cause the plant to
develop suckers near theground. Some varieties produce more suckers
thanothers.
The inflorescence is a terminal panicle. The flowersare about 5 cm
long and light pink in color, although

 Page 3
varieties producing white and carmine-red blossoms are known. The
flower color of N. tabacum is usually pink; that of N. rustica yellow to
greenish yellow. The flower is borne on a short stem and has a five-
cleft calyx, from which emerges the slender corolla tube. The latter is
much longer than the calyx and usually expands at the top into a five-
lobed limb. The stamens, five in number, are attached to the corolla
tube and each consists of a long, slender filament and an ovalshaped
anther that splits lengthwise in discharging its pollen. The pistil is
composed of a swollen basal element, the ovary that contains the
ovules arranged on a fleshy placenta, a long slender style arising
above the ovary, and a more or less two-lobed stigma. Normally some,
but not all, of the stamens are of about the same height as the stigma.
It is apparent that the structure of the flower is such as to favor self-
fertilization. At maturity, the ovary is generally enlarged and becomes
the two-celled fruit or seed pod bearing numerous seeds.
The tobacco plant possesses an extensive but comparatively shallow
system of fibrous roots that usually affords adequate support although
the plant may be toppled in a strong wind, especially if it bears a
heavy top, such as a full-size flower head. Most of these roots develop
adventitiously from the portion of the main stem buried during
transplanting.
b
Chemical Composition
Several review articles and publications have focused on the role of
leaf composition to flavor and aroma of the smoke (Davis, 1976;
Enzell, 1976; Leffingwell, 1976). Many essential characteristics of
tobacco leaf, particularly alkaloids and flavor components, can be
traced to the progenitor species (Legg & Collins, 1974). For example,
in a study with SC 58 alkaloid isolines, genetic control of fatty acids
and alkaloids was demonstrated as shown in Table 1.2 (Weeks, 1985).
The precursors of norisoprenoids are carotenoids, labdanoids and
thunberganoids. These compounds are
Table 1.2 Fatty acids and alkaloids
associated with SC 58 alkaloid
isolines.
Alkaloid Alkaloids Total fatty acids
isolines (%) C14-20 (mg/g)
aabb 0.23 5.35
aaBB 1.50 3.75
AAbb 2.20 4.40
AABB 4.06 4.01

found in the three wild species mentioned above (Enzell, 1976;

Leffingwell, 1976; Enzell, 1988).
Among the 1500 entries in the USDA germplasm inventory, there is a
wide range of variations in chemical constituents and smoke delivery
even in plants grown at the same location under similar cultural
practices, as shown in Table 1.3 for flue-cured and burley crop
production systems (Chaplin, 1980).
Table 1.3 Variations in certain chemical
constituents of approximately 1500
tobacco introductions and cultivars
produced at one location under flue-cured
and burley cultures.
Constituents Range
Flue-cured culture
Tar/cigt (mg)1calculated 15.5837.59
Reducing sugars (%) 0.8022.20
Total nitrogen (%) 2.305.25
Total alkaloids (%) 0.207.87
Petroleum ether extract 6.5115.30
(%)
Wax (%) 0.221.83
Holocellulose (%) 22.0443.08
Total phelols (%) 0.205.99
Burley culture
Nitrates as N (%) 0.011.18
Total nitrogen (%) 2.154.85
Total alkaloids (%) 0.025.69
Total volatile bases (%) 0.261.33
Petroleum ether extract 1.117.93
(%)
Cellulose (%) 3.4415.14
P (%) 0.140.45
K (%) 1.055.40
Ca (%) 1.3510.35
Sterols (mg/g)2 1.174.39
1 Estimation of tar by multiple regression
technique.
2 Based on 75 selected samples from the
approximately 1500 Tls and cultivars.

Genetic control of leaf quality is well recognized (Smeeton, 1987).

Studies on levels of solanesol, sterols, fatty acids, and lipids among
various cultures (Chaplin, 1980) are also reported, as shown in Tables
1.4, 1.5, and 1.6 respectively. All these data clearly demonstrate the
wide genetic variation that exists for leaf chemical components and
indicate that there are considerable opportunities for manipulation by
plant breeders toward a desired direction. Recent progress in the
production of transgenics offers an even greater potential for variety
development. A number of issues, including consumer and industry
acceptance, must be

 Page 4
Table 1.4 Comparison of the levels
of solanesol of normal green
fluecured tobacco cultivars and
their pale yellow selections.
Cultivar or selection Solanesol (%)
VA 115 1.15
Average pale yellow 0.99
Coker 139 1.11
Average pale yellow 1.03
NC 2326 1.34
Average pale yellow 1.27
NC 95 1.30
Average pale yellow 1.19
Coker 187 Hicks 1.21
Average pale yellow 1.03
Average cultivar 1.22
Average pale yellow 1.10
Average of pale yellow selections in
each family.

Table 1.5 Sterols and total alkaloids of

fluecured tobacco cultivars.
Cultivars Total Total
sterols (%) alkaloids
(%)
Coker 319 0.313 2.24
VA 115 0.263 2.60
NC 95 0.259 2.72
McNair 12 0.251 2.62
Speight G-7 0.248 2.64
Hicks broadleaf 0.241 2.69
NC 2326 0.231 2.62
McNair 30 0.224 2.53
Reams 266 0.193 1.69
Golden wilt 0.172 2.75
Cultivar LSD 0.020 0.21
(0.05)
(0.01) 0.27 0.29

Table 1.6 Fatty acids and lipid residues in

four fluecured cultivars.
Cultivar Total Fatty Lipid
alkaloids acids residues
(%) (mg/g) (mg/g)
SC 58 3.58 5.7 81.6
LN 38 0.10 7.2 81.1
NC 95 2.87 5.7 72.5
Coker 1.37 4.8 69.9
139
LSD 0.28 0.5 2.2
(0.05)
(0.01) 0.40 0.7 3.2

resolved prior to the use of transgenics becoming a reality.

Leaves from different stalk positions on the same plant differ
chemically (Rogers & Mitchem, 1976). For example, total nitrogen
and nicotine contents increase with higher node position, whereas
reducing sugars are greatest in leaves from the middle stalk (Tables
1.7 and 1.8). By varying cultural practices such as plant spacing,
topping and degree of sucker control, the concentrations of leaf
compounds can also be changed, thus providing a further opportunity
to manipulate leaf characteristics (Papenfus & Quin, 1984).
Table 1.7 Average analyses of
nicotine by stalk position.
Type Stalk Nicotine
position (%)
FluecuredLower 1/3 1.87
FluecuredMiddle 1/3 2.65
FluecuredUpper 1/3 3.26
Burley Lower 1/3 2.14
Burley Middle 1/3 3.00
Burley Upper 1/3 3.65
Oriental Composite 0.95
Stem- Composite 0.85
sheet

c
Pest Resistance
Many diseases and insect problems occur during tobacco production,
some even at the storage stage of finished products (Tso, 1972; Tso,
1990). Severe infection and damage by pests such as insects, mites
and nematodes can affect normal growth which usually results in
smaller yields and inferior quality (Pirone, 1979).
The effects of pests and diseases and their control have been well
studied and are discussed in detail in later chapters of this CORESTA
monograph.
Although control by pesticides is feasible in most instances, selection
and breeding for disease and pest resistance is a major scientific
occupation in the industry. The use of resistant varieties is an
important component of integrated pest management. In addition to
being economically and environmentally sound, it minimizes the
possible influence of pesticides on leaf and, therefore, smoke quality.
In this context, pesticide residues are assiduously monitored by
practically all major leaf suppliers and manufacturers (Sheets &
Leidy, 198798).
It is important to examine pest resistance of tobacco


 Page 5
Table 1.8 Chemical analysis of a typical flue-cured tobacco by plant
position.
Priming Total Amino Nicotine Reducing WSA1 pH
nitrogen (%) nitrogen (%) (%) sugars (%)
7 2.31 0.262 3.89 9.8 5.15 5.07
6 2.21 0.168 3.35 15.5 4.97 5.10
5 1.78 0.102 2.47 20.3 4.21 5.16
4 1.55 0.084 1.82 22.7 3.75 5.22
3 1.48 0.092 1.53 20.6 3.42 5.30
2 1.69 0.134 1.40 16.3 3.42 5.35
1 1.77 0.184 1.28 10.6 3.47 5.52
Weighted 1.77 0.124 2.15 18.3 3.85 5.25
average
1 ml of 0.1 N NaOH to neutralize 1 g tobacco.

from a genetic point of view. Certain plants are susceptible and others
are resistant because of their differences in genetic make-up.
Constituents important to tobacco quality include nitrogenous
components, lipids, sugars, phenolics and terpenoids, and these have
been shown to be quantitatively and qualitatively altered in diseased
plants. Phenolics and terpenoids may have a role in restricting the
spread of pathogens in plant tissue (Baldwin, 1988). Leaf surface
chemistry, including such compounds as the duvanes, is of importance
to insect damage and resistance (Gwynn, et al., 1983; Severson, et al.,
1984; Severson, et al., 1985; Shah & Chakraborty, 1985; Jackson, et
al., 1987).
Although the various mechanisms of pest resistance are not yet well
established, it is generally believed that breeding tobacco plants for
pest resistance is the most effective and environmentally compatible
approach for economic and safer tobacco production. Conventional
plant breeding techniques remain an effective means of increasing
plant tolerance and resistance to pests as illustrated during the last 60
years of growth in the tobacco industry (Kehr & Smith, 1954; Legg &
Collins, 1974; Chaplin et al., 1976). Genetic engineering techniques
have been used to create plants which are protected from attack by
larvae of the tobacco hornworm (Manduca sexta). Subfragments of
the gene encoding the Bacillus thuringiensis subsp. berlines (B.
thuringiensis subsp. thuringiensis) delta-toxin were cloned and
inserted into plants using vectors based on the tumorinducing plasmid
of Agrobacterium tumefaciens (Gheysen, et al., 1987; Shields, 1987).
The regenerated plants expressed the foreign genes and produced
sufficient insecticidal protein to protect them from larval damage. A
gene encoding a trypsin inhibitor in Vigna unguiculata and known to
give some measure of field resistance to insect pests was transferred
to Nicotiana tabacum cv Samsun NN by means of transformation of
leaf discs using an Agrobacterium tumefaciens Ti plasmid binary
vector (Hilder, et al., 1987; Tso, 1972). When sets of clonal plants
derived from the transformed plantlets were exposed to H. virescens,
the percentage leaf area eaten was as low as 21% versus control plants
which were reduced to their stalks by the insect larvae. Again, as
stated in the previous section, the use of transgenics has not been
accepted by the tobacco industry and some consumers.
d
Physiological Responses
Many physiological disorders examined in tobacco plants are due to
responses to abnormal environmental conditions which triggered their
genetic expression and are not diseases induced by pathogens. The
most prominent physiological disorders include genetic tumors,
pollutant effects and frenching (Tso, 1972). Nutrition deficiency
induced disorders are not related to genetic make-up, although there
are degrees of sensitivity between types and varieties.
Genetic Tumors
Certain Nicotiana hybrids produce teratoid proliferations of a genetic
origin. Such tumors may grow spontaneously on stems, roots, leaves
or flower systems on tumorous F1 progeny or on only part of a tissue.
For example, Nicotiana species within the section Alatae produce
tumorous hybrids when combined with certain species in other
subgeneric sections (Kostoff, 1943; Kehr & Smith, 1954; Naf, 1958).
The species involved in the production of tumorous interspecific
Nicotiana hybrids can be separated into two groups (Naf, 1958) one
called the 'plus' group, and the other the 'minus'

 Page 6
group. Hybrids among plus species or hybrids among minus species
are free from tumors. The critical contribution of the 'plus' parent to
tumor production in the hybrid differs from the critical contribution of
the 'minus' parent. Based on the above hypothesis, four new hybrids
have been produced: N. rustica × N. forgetiana, N. rustica × N.
bonariensis, N. suaveolens × N. bonariensis, and N. suaveolens × N.
longiflora. There is evidence (Johnson, et al., 1974) that tumor
formation is controlled by 'conventional' genes that show segregation,
linkage, and mutation.
Wounding and radiation can induce tumor formation (Hagen, et al.,
1961; Hagen & Gunckel, 1962). Likewise, plant growth hormones
may either inhibit or accelerate the expression of tumor formation.
Many chemicals have been tested for their effect on tumor formation
and may be positive or negative (Tso & Umbarger, 1961; Tso & Burk,
1962). Such genetic tumors do not occur in cultivated tobacco plants.
It must be noted that tumors developed among certain hybrids of
Nicotiana species are totally different from those of experimental
animal tumors and should not be confused.
Changes in chemical composition of tumorous Nicotiana plants are
the subject of many academic investigations. The amphidiploid hybrid
of N. glauca × N. langsdorffii has a higher content of certain free
amino acids than the parent species, suggesting that certain free amino
acids may act as cecidogenic agents. In an examination of alkaloids,
sugars, organic acids and amino acids of N. glauca, N. langsdorffii
and tumorous and visibly nontumorous tissues of their F1 hybrids,
anabasine and nicotine, respectively, were found to be the principal
alkaloids in the parent species (Tso, et al., 1967). In the F1 hybrids,
however, nornicotine was the principal alkaloid of the tumorous
tissue, and nicotine, while low in concentration, was the principal
alkaloid of the nontumorous tissue. In another study, a sharp increase
of scopolin and the new formation of scopoletin in the tumorous tissue
of Nicotiana hybrids was observed (Tso, et al., 1964) as compared
with parental material. These new compounds are possibly related to
the formation of tumors in the F1 hybrids.
Air Pollutants
Many factors contribute to air pollution, but the most significant
source is from energy conversion. Energy conversion products that are
considered pollutants may include water, carbon dioxide, carbon
monoxide, sulfur dioxide, sulfuric acid, hydrogen sulfide, nitric oxide,
nitrogen dioxides, hydrogen fluoroxide, ethylene, ozone, aldehydes,
soot and hydrocarbons. Other materials such as radioactive materials
and many other organic and inorganic wastes are also present in our
environment as pollutants and may have a significant effect on
tobacco production and quality in certain localities, e.g. herbicidal
contamination of a tobacco field from a nearby field.
Effects of air pollutants on agricultural yield, including tobacco, have
been recognized since the 1950s (Roberts, 1984). Tobacco, indeed,
was among the first agronomic crops to exhibit leaf disorder following
prolonged exposure to air pollution and served as an indicator plant in
studies on the effect of air pollution on plant growth.
In general, air pollution either causes injuries that are visible as small
patches of brown necrotic areas covering the upper surface of leaves
or it causes damage that is restricted to leaf margins or tips. These
visible injuries are the result of localized death of tissues 24 hours
after exposure to air pollutants. Invisible injuries on the other hand
may involve impairments of the photosynthetic system in the
mesophyll cells, a change in the stomatal control of gas exchange or
changes in enzymatic activities (peroxidase, etc.). Air pollutants are
collectively called 'smog', which is a mixture of gaseous oxidants
formed in the atmosphere resulting from photochemical reactions of
which ozone is a major component. Other components present in
smog that are known to cause damage to plants are peroxyacetyl
nitrate (PAN), aldehydes, the product of oxidation of hydrocarbon
fragments, and SO2 (which upon oxidation by peroxy radicals, forms
SO3 and, when combined with water, forms sulfuric acid mist, a
component of acid rain).
Tobacco weather fleck is a visible injury of a leaf; it is a physiological
disorder of the leaf which may be caused by either high levels of
ozone or acid rain (Dean, 1963; Rathier & Frink, 1984). Weather fleck
is characterized by small necrotic lesions on the leaf surface, usually
0.20.5 cm in diameter. In general, a high incidence of weather fleck
lowers the yield, reduces the value of tobacco and may cause earlier
flowering, lower total alkaloids, taller plants and longer internodes
(Aycock, 1975). The recognition of the heritable trait of weather fleck
susceptibility has contributed to the development of breeding
programs utilizing hybridization and selection of pure lines in an
effort to increase the level of weather fleck resistance in tobacco. The
degree of weather fleck varies among locations, years and tobacco
varieties.

 Page 7
Frenching
Frenching is a noninfectious physiological disorder of tobacco. It
usually appears as a network chlorosis of apical leaves, followed by
formation of progressively narrower leaves. In extreme cases there is
very limited expansion of lamina in younger leaves (strapping) or
stalk elongation. Many Nicotiana species are susceptible to frenching
(Steinberg & Tso, 1958). Severe symptoms have been observed in N.
alata, N. langsdorffii, N. longiflora, N. rustica, N. sanderae, N.
sylvestris, and many varieties of N. tabacum. Steinberg conducted a
series of studies (Steinberg, 1947; Steinberg, 1950; Steinberg, 1952)
and concluded that frenching is caused by an organic toxin produced
by Bacillus cereus Frankland and Frankland. It was suggested that
diffusates from B. cereus and perhaps other soil bacteria may be the
cause of frenching of tobacco in the field. Increased populations of the
bacillus organisms have been found in adjacent soil and rhizospheres
of frenched plants (Steinberg, 1951). However, leaves of frenched
plants are high in L-isoleucine (Steinberg, et al., 1950) and free L-
isoleucine, which caused frenching in aseptic culture (Steinberg,
1949). Its capacity in this respect is exceeded fourfold by L-
alloisoleucine (Steinberg, 1949). However, leaves of frenched plants
are high in L-isoleucine (Steinberg, et al., 1950) and the frenching
phenomena are evidently a combination of biological, chemical and
genetic effects.
In summary, it is generally recognized that there are opportunities for
tobacco improvement via genetic manipulation. In addition to
conventional breeding, molecular biology and genetic engineering
offer new and additional tools to meet agricultural needs in producing
better tobacco. However, much has yet to be learned and accepted.
The successful application of gene transfer technology depends upon
breakthroughs in the understanding of gene expression and regulation,
as well as increased knowledge in the regulation of cellular processes
(metabolism).
Fnvironmental Influences
a
Soil, Moisture and Air
Although tobacco can be grown in a wide range of soils from sands to
heavy clays, each type has reasonably specific soil requirements to
produce optimal quality. Tobacco has a very active root system which
is required to support the development of an enormous leaf area
within a short period of time. Adequate soil aeration, water and
nutrient supply are three major requirements for maximum leaf
expansion. It is evident that to ensure thorough soil aeration and
drainage at all times, a relatively open, loose soil structure is essential.
On the other hand, for maximum foliage expansion it is essential that
the tissues be fully turgid at all times, and this, in turn, requires an
abundant moisture supply in the soil. The need for a balanced and
adequate supply of plant-food elements to support this growth is
obvious.
In the case of cigar wrapper leaf, for example, when the above
mentioned conditions are fulfilled, the plant produces a relatively
large, broad and extremely thin leaf with a fine, open texture of low
density. Such leaf is of light color and when cured has good
combustibility. It is the thinnest and lightest commercial type of leaf
produced.
The required combination of thorough soil aeration and liberal
moisture supply is most likely to occur in sandssandy loam soils
which are used almost exclusively in the production of flue-cured leaf.
Heavier soils are more suited to the production of cigar filler, dark air-
cured and dark fire-cured types where the resulting leaves are thicker,
heavier, darker colored, more oily, stronger and slower burning. In
soils for these tobacco types, aeration can become more restricted and
the balance between aeration and available moisture more critical.
Burley tobacco furnishes an outstanding exception to the general rule.
Burley tobacco, whose leaf is relatively thin, light in weight and color
and has good burning qualities, is grown most successfully on highly
fertile but quite heavy silt loam soils. This result, however, is due
essentially to inherent genetic/physiological characteristics of the
'white stem' burley varieties. In recent years, many of the cigarette
manufacturers have tended to purchase a fuller bodied, darker burley
leaf. Other types grown on the same heavier soils will produce only
the heavy, dark type of leaf.
Oriental tobacco requires thin, rather infertile soils with a low
nitrogen content. Such soil generally is located on the lower mountain
slopes or in the foothills. Table 1.9 shows several well-known tobacco
types and their most productive soil types in the USA and Cuba
(Akehurst, 1981).
Imbalanced fertilization and soil acidity (pH) are known to be
associated with many diseases of tobacco (Collins & Hawks, 1993)
and the availability of certain nutrients to the tobacco plant (Komatsu,
1954).
Soil, moisture and air are three separate identities which interact
closely. Soil, in addition to providing support for a plant, must provide
an adequate moisture

 Page 8
Table 1.9 Soil types in the USA and
Cuba and the tobacco types to which
they are best suited for production.
Soil type Tobacco
type
Sandy loam to fine sandy Flue-cured
loam
Silt loam Burley
Fine sandy loam Maryland
Cigar
Loam Wrapper
Silt loam Filler (PA)
Fine sandy loam Filler
(Cuba)
Sandy loam Binder
Clay loam, silt loam Dark fire-
cured
Clay sand topsoil and Oriental
chalky subsoil
Source: Tso (1990).

supply as well as air for oxygen. Soil structure should allow free
moisture movement and air penetration for best growth and
development. Such conditions are typical of lighter soils in the sand-
loam range. Plants need water to transport nutrients and to maintain
full turgidity, which is a necessary condition for maximum expansion
and minimum thickening of the leaf. On the other hand, excessive
water causes leaching of nutrients from the soil-root zone. More
importantly, if a tobacco field is flooded for a period as short as 4
hours, the crop may be damaged. In one study, intentional flooding for
48 hours after the first priming of flue-cured tobacco reduced the yield
less than 15% (Felipe & Long, 1988). Time within the growth cycle,
soil characteristics and extent of flooding impact the amount of
damage. Lack of aeration resulting from prolonged flooding causes
injury and death of the roots. Such an injury is aggravated by the
activity of microorganisms which destroy the roots and plug the
xylem of the stem base. Also, it is likely that toxic substances
produced by microorganisms in the dying root cells contribute to
damage of the root. In addition to water and nutrients, growing shoots
are dependent upon functional root systems for growth promoting
substances (Tso, 1972).
The amount and timing of the water supply are critical factors for
success or failure of a tobacco crop. Although a tobacco plant can
tolerate moderate droughts, extended drought can seriously affect
growth or interrupt normal physiological processes to a point of no
return.
Many farmers irrigate to supplement rainfall. However, after the
seedlings are transplanted into the field, a short period of dry weather
encourages root growth and extension. Thereafter, during the period
of rapid growth and development which generally occurs from the
fourth to the eighth week after transplanting, an adequate water supply
is essential. Even during the harvesting phase of tobacco to be primed,
if the weather is too dry and tobacco leaves lack moisture,
supplementary irrigation may be necessary to maintain the quality of
the cured leaf (Long & Weybrew, 1982). However, over-irrigation at
this stage also induces damage. A limited moisture supply is needed
for Oriental tobaccos in comparison with flue-cured types. As a result,
Oriental tobacco is smaller in size and higher in aromatic constituents
and resins than flue-cured.
In flue-cured tobacco, soil moisture tension was found to influence the
chemical composition of the leaves. Low tension (high moisture
content) gave lower concentrations in the cured leaves of nicotine,
total nitrogen, calcium oxide (CaO), and magnesium oxide (MgO),
but higher concentrations of sugars, as well as better burning
characteristics and higher yields. Conversely, high moisture stress
reduced the proportion of carbohydrates to nitrogen compounds and
favored the production of a thicker, coarse-textured tobacco.
Calculated on a nitrogen compounds/ha basis, there is seldom, if ever,
a significant increase in nitrogen compounds. However, there is a
significant reduction in the carbohydrate fraction. Also, dry weather
increased the proportion of nitrogen present as amino acids, raised the
petroleum-ether extractables (gums, oils and resins) and increased
carbohydrate oxidation to acids (Tso, 1972). It can be considered that
soil, nutrients, water and air interact closely as principal factors
determining the characteristics of a given tobacco crop (Weybrew, et
al., 1983). Those factors have to complement each other to achieve
the best product.
b
Nutrition
Desirable leaf quality or 'usability' requires a subtle balance of
chemical and physical properties, involving visible, physical and
chemical criteria. All these quality factors are affected, directly or
indirectly, by nutrition (Tso, 1972).
Mineral nutrition plays an important role in producing high quality
leaf tobacco. Nutritional disorders in tobacco (Nicotiana tabacum L.)
are basically physiological and biochemical phenomena induced by
deficiency, excess or interaction among certain elements under
specific environmental conditions. Such disorders are generally
characterized by visible symptoms in the plant which reflect the
imbalance of its metabolic system.

 Page 9
Advances in analytical technology and instrumentation have made it
possible to detect very minute quantities of practically all chemical
elements in leaf tobacco (Tso, 1972; Green & Rodgman, 1996). The
presence of any particular element in the plant does not necessarily
mean it is essential for normal growth and development. In fact, many
unwanted elements (e.g. cobalt (Co), nickel (Ni), copper (Cu), mercury
(Hg), thallium (Tl), silver (Ag) and cadmium (Cd)), if present in
sufficient quantities, demonstrate their toxicity at the cellular level
(Siegel, 1977).
Due to its sensitivity, tobacco is one of the most extensively used crop
plants in nutrition studies, especially those dealing with nutrient
deficiencies. Many factors influence disorders due to nutrient amounts,
timing, forms, temperature, soil acidity, transfer within plant, plant age
and stalk position.
A deficiency of any element essential for the normal metabolic
processes of tobacco plants results in either visual or chemical
abnormalities, or both. Extensive studies have been conducted in this
area most pioneer work being done by McMurtrey (1933). A reduction
in growth is usually accompanied by typical characteristic
abnormalities which provide a reliable basis for distinguishing one
deficiency from another. The symptoms produced by deficiencies fall
broadly into two groups. One group includes those resulting from lack
of nitrogen, phosphorus, potassium and magnesium, which apparently
are readily mobile in the plant. The symptoms are initially localized on
the older or lower leaves, but later spread throughout the whole plant.
Another group of symptoms consists of those caused by lack of
calcium, boron, manganese, sulphur and iron, which, from symptom
manifestations, are relatively immobile and are localized on the
terminal growth consisting of upper or bud leaves. A detailed
examination key was provided by McMurtrey (1933) for distinguishing
such deficiencies, as shown in Table 1.10. Symptoms observed in field
conditions are similar to those in solution culture (McMurtrey, 1939).
Table 1.10 Key to mineral deficiency symptoms on tobacco.
1. Effects localized on older or lower leaves or more or less general on (Group 1 )
whole plant.
1.1 Local: occurring as mottling or chlorosis with or without necrotic
sporing of lower leaves; little or no drying up of lower leaves.
1.1.1 Lower leaves curved or cupped under with yellowish Potassium
mottling at tips and margins. Necrotic spots at tips and margins.
1.1.2 Lower leaves chlorotic between the principal veins at tips,
and margins of a light-green to white color. Typically, there are no
necrotic spots. Magnesium
1.2 General: also yellowing and drying or 'firing' of lower leaves.
1.2.1 Plant light green, lower leaves yellow, drying to light-brown Nitrogen
color.
1.2.2 Plants dark green, leaves narrow in proportion to length; Phosphorus
plants immature.
2. Effects localized on terminal growth, consisting of upper and bud (Group 2)
leaves.
2.1 Dieback involving the terminal bud, which is preceded by
peculiar distortions and necrosis at the tips or base of young leaves
making up the terminal growth.
2.1.1 Young leaves making up the terminal bud first light green,
followed by a typical hooking downward at tips, followed by
Calcium
necrosis, so that if later growth takes place, tips and margins of the
upper leaves are missing.
2.1.2 Young leaves constricted and light green at base, followed by
more or less decomposition at leaf base: if later growth takes
Boron
place, leaves show a twisted or distorted development; broken
leaves show blackening of vascular tissue.
2.2 Terminal bud remains alive, chlorosis of upper or bud leaves,
with or without necrotic spots, veins light or dark green.
2.2.1 Young leaves with necrotic spots scattered over chlorotic Manganese
leaf, smallest veins tend to remain green, producing a checkered
effect.
2.2.2 Young leaves without necrotic spots, chlorosis does or does
 not involve veins so as to make them dark or light green in color.
2.2.3 Young leaves with veins of a light-green color or of same Sulfur
shade as intervein tissue. Color light green, never white or yellow.
Lower leaves do not dry up.
2.2.4 Young leaves chlorotic, principal veins characteristically Iron
darker green than tissue between the veins. When the veins lose
their color, all of the leaf tissue is white or yellow.

 Page 10
Subsequent studies (Takahashi & Yoshida, 1957) reported distinctive
symptoms of the deficiency of other minor elements. The symptoms
of molybdenum (Mo) deficiency, like those of zinc (Zn) and Cu, first
appeared on the middle leaves. As in the case of Cu deficiency, the
dead leaf areas became almost white. In Zn deficiency, the dead areas
were light to dark brown with dark irregular rings and granules.
Studies on the effects of mineral excess are fewer than those on
mineral deficiency, especially in the area of major elements. Toxicity
due to excess of minor elements is, however, more frequently
reported.
Excess nitrogen delays flowering, maturation and ripening by
prolonging the vegetative phase through extended dominance of
protein metabolism. Nitrogen requirements of tobacco varies with
tobacco types, and water supply is one of the major factors affecting
nitrogen utilization. As mentioned previously, excessive delay in
maturation of leaves may also induce higher incidence of disease.
Although a high nitrogen supply generally increases yield, it is
necessary to achieve a balance between maximum yield and optimal
quality.
Moderate excess of phosphorus usually causes no marked effect on
tobacco. Extremely high levels of phosphorus can reduce yields and
result in narrow thick leaves. High potassium application tends to
delay leaf maturity, but improves rate of burn. In general farm
practice, the amount of potassium used often exceeds normal
requirements.
High chloride (Cl, used generically in this monograph to refer to the
chloride anion, , which is absorbed by the roots. Previous literature
referred to Cl as chlorine which is incorrect since chlorine, Cl2, is a
gas) concentrations affect both growth and quality, resulting in leaf
that is very brittle and thickened. Margins curl upward and the leaf
has a distinctive sleek, glabrous appearance (McMurtrey, 1939).
Leaves with excessive Cl have high equilibrium moisture contents, i.e.
they are very hygroscopic and have low or even no fire-holding
capacity. Such tobaccos are considered to be poor quality by the
cigarette manufacturers. The use of fertilizers that contain high levels
of Cl is discouraged by this part of the industry.
In summary, most nutritional elements are present in tobacco soils.
Some are present in abundant quantities, and others in minute
amounts. These elements vary in chemical and physical properties.
Soil pH greatly affects their availability to plants. Under the
conditions for tobacco production, many events may lead to
interactions among these elements and thus affect growth and
development of the tobacco plant.
c
Temperature and General Climate
Tobacco is a native of the subtropical zone. For economic reasons, it
is now being produced commercially in almost every corner of the
earth, between latitudes 55°N and 40°S, although the best locations
are generally in a much narrower belt. The most important basic
requirements are at least 120 (preferably 140) frost-free days for field
growth, adequate water for the particular type, sufficiently high
temperatures and sunlight for fundamental biochemical and
physiological processes. Also, the incidence of certain tobacco
diseases, such as blue mold, is significantly influenced by moisture
and temperature.
Although certain tobacco types can tolerate short durations of extreme
temperatures, prolonged exposure causes serious stress, sometimes
permanent damage to the plants. Generally, in the early stage (1 to 4
weeks), following transplanting, cold temperatures retard root
extension and therefore affect young plant establishment. The
following 4 weeks (5 to 8) are critical for leaf growth and expansion.
In the case of flue-cured tobacco, minimum and maximum
temperatures of 18° to 22°C and day temperatures of 28° to 32°C are
considered ideal. However, most locations are not so favorable and
yet produce high yields of acceptable quality leaf.
Generally, for all tobacco types, temperatures below 13°C are not
desirable, particularly when combined with wet weather without much
sunshine. Average temperatures around 27°C in a warm climate with
abundant sunshine provide good growth conditions under which crops
can reach maturity in 80 to 90 days from transplanting. In colder
climates, it may take 100 to 120 days. Inadequate sunshine results in
poor growth, and leaves can hardly reach true maturity, resulting in
poor quality tobacco.
During the growth period, sunlight and moderately high temperatures
are essential for dry-matter production and accumulation as well as
related metabolic changes. Even at time of harvest, too low a
temperature limits the normal biochemical process for successful
ripening and thence curing, especially for the air-cured types.
Night temperature profoundly affects total amounts and proportions of
cell walls, particulate proteins, soluble cytoplasmic proteins, growth
habit and rate, time of flowering, and final yield (Tso, 1990). A low
temperature regime before transplanting can promote premature floral
initiation and, also, it promotes certain mineral deficiency symptoms,
a factor probably associated with genetically inherited enzyme
activities of

 Page 11
certain varieties. Temperature significantly affects the availability of
nutrients to tobacco plants. For example, uptake of phosphorus (P)
decreases as temperature falls from 35° to 10°C (McEvoy, 1960).
Insufficient supplies of P result in general plant retardation, narrow
leaves, delayed flowering and abnormal 'maturation' (Komatsu, 1954).
Further, it induces physiological diseases such as weather fleck or
spotting (McCants & Woltz, 1967).
d
Day Length
Wet, cold weather frequently induces early flowering of the young
seedling. A cold, short day is often associated with premature early
flowering of many Nicotiana plants.
In 1918, two scientists, W.W. Garner and H.A. Allard of the USDA,
tried to induce flowering in Maryland Mammoth tobacco, a plant
mutation. They were unsuccessful throughout the year until they
moved the plants into a greenhouse to protect them from winter frost.
The plants flowered about Christmas time. This observation led to a
new era of plant science. They discovered the fundamental principle
that the relative length of day and night controls flowering. They
named the phenomenon photoperiodism. Short-day plants flower
when days are short and nights are long, but do not flower under other
conditions of day length. Long-day plants flower only when the days
are long and nights are short. Still others are day-neutral, or show no
preference as to day length. The historical 'dark houses' in Arlington,
Virginia, Garner and Allard used for tobacco research on day-length
are now the location of the Pentagon Building.
Most Nicotiana species are day-neutral (Chaplin & Burk, 1979).
Premature floral initiation results in fewer potentially harvestable
leaves and can, therefore, have serious commercial consequences.
Photoperiod is not solely responsible. Also, temperature is implicated
and cool temperatures combined with short days are particularly
damaging. Furthermore, temperature alone can also affect leaf number
in a noninductive photoperiod (Thomas, et al., 1975). Most
commercial cultivars behave as qualitative (facultative) short-day
plants, forming their maximum leaf number in photoperiods of more
than 11 hours and mean daily temperatures of at least 18°C
(Hopkinson, 1969; Kasperbauer, 1969). In research involving
interspecific hybridization, scientists have to bring species of different
day lengths to flower at the same time in order to make hybridization.
It was observed that there are interactions between day length and
temperature, such as warm-long, cold-long, warm-short and cold-short
to induce flowering.
In addition to day length, light quality has been the subject of many
studies. Light quality affects leaf composition as well as physical
shape (Tso, 1972).
Agronomics (Human Intervention)
a
Seeds and Seedlings
Fully developed, healthy, clean seeds have the greatest potential for
transferring genetic information from one generation to another.
Tobacco seeds are extremely small in size. There are about
1000013000 seeds per gram. Each flue-cured tobacco plant may
produce 12 to 15 grams of seed, about 150 000 seeds, sufficient for
about 250 sq. meters of conventional seedbed for widely grown types
such as burley and flue-cured. Such an area will normally provide
sufficient, good quality seedlings for about 3 ha of field crop in the
case of these types (Chaplin, et al., 1976). The length of the seedling
phase varies according to production system and environment. This
variation is from 6 to 8 weeks in heated greenhouses and for outside
seedbeds grown in the low altitude tropics to about 12 weeks for open
beds in cooler environments, such as for the main planting region of
Zimbabwe. Because of the practice of seedling clipping, 1 gram of
seed is now used to plant 585 to 699 sq. metres of seedbed. Seed from
one plant may be used to plant at least 6.5 ha of tobacco.
Tobacco seeds maintain high viability under proper storage conditions
for as long as 25 years in air-tight containers, either refrigerated or
desiccated or both. Free access of air appears to be unfavorable to
maintain viability for more than 15 years. It is common practice to
store tobacco seeds at a low moisture content (approximately 7%) at a
temperature below 21°C in an air-tight container.
The seed of N. tabacum contains little, if any, starch. There is starch in
the ovule and it develops very early in the hypocotyl of the young
seedling. Both endosperm and embryo contain abundant but
chemically different proteins. The presence of alkaloids or related
pyridyl compounds in tobacco seed is a question of much
disagreement. Most investigators have failed to recognize chemical
variations in the seed that result from differences in variety or species,
and from stage of seed development or degree of seed maturity.
Seed size influences germination rate and seedling

 Page 12
vigor, which are important factors in innovative starting procedures
such as seeding directly in the field or in transplantable containers.
Heavy seeds germinate earlier, in higher percentages, and over a
shorter period of time than do the lightweight seeds. Seeds that
germinate on the same date produce seedlings that do not differ
significantly in size of transplants or in subsequent field growth and
development, including leaf yield.
Essential factors for a good seedbed location include maximum
natural warmth, southern slope with free exposure to sunlight,
effective windbreaks, sufficient surface and underground drainage,
fertile soil with desirable texture, tilth and moisture holding power
and the bed should be some distance from the curing barns to avoid
contamination by diseases.
Clipping to control seedling growth is frequently practiced. It is a
useful procedure to control plant growth, improve uniformity and
better manage transplanting schedules (Miner, et al., 1983; Suggs, et
al., 1988). Undercutting, in which a blade is passed through the soil
about 7 to 10 cm below the surface, is not a widely accepted practice
in some countries. It has been shown to increase root growth, which is
important for transplant establishment, and retard shoot growth so that
transplanting can be extended over a longer period of time. Clipping
will accomplish the same goals.
Transplanting seedlings to the field by pulling (bareroot) or by using
intact roots (soil plug, pot, etc.) has been the focus of some research
(Suggs & Mohapatra, 1988). Intact-root transplants are intuitively
attractive because transplanting shock is decreased. Also, because
intact-root plants are grown in an orderly array in cells or other
containers, they can be automatically singulated and fed into a
transplanter. Intact-root transplants are often expected to yield more
than bareroot transplants.
Hardening of seedlings, which involves one or more cycles of
withholding water until soil moisture potential is close to wilting point
and then rewatering, decreases transplanting shock and so improves
field growth in countries with hot, dry environments at transplanting
time (Papenfus & Quin, 1984).
Optimal seedling size varies from one situation to the next, depending
on factors such as transplanting method (by machine or manually),
soil moisture status, evaporative demand and whether or not it is
necessary to bury the roots below a herbicide treated layer of soil.
Large seedlings per se, especially those with good root systems and
leaf canopies, establish and perform better than smaller ones
(Papenfus, 1987).
b
Plant Populations and Leaf Production
Among various tobacco types, there is an extremely wide range of
spacing variation. The effect of spacing (plant populations) on leaf
usability and quality is far greater than yield (Whitfield & Connor,
1980; Papenfus, 1987). Plant populations range from 8000 to 30 000
per ha for broad leaf types. Within this range, the widest spacing
applies to dark air-cured and fire-cured types and the closest to shade-
grown cigar wrapper. However, Oriental tobaccos may be planted at
populations as large as 150 000 per ha. Closer spacing of plants
results in reduction in size, body, thickness and weight per unit area of
the leaf. Also, seed production is affected.
In a study of the architecture of individual plants in the flue-cured
tobacco field based on foliage displayed, it was found that in mature
crops, the foliage extended further into the inter-row space than into
the space occupied by neighboring plants in the row. Mean leaf angle
was 40° and elevation distributions were remarkably similar
throughout growth and development. Foliage inclination consistently
decreased with depth in the canopy.
The chemical composition in leaf tobacco is a function principally of
plant and leaf population (Weybrew & Woltz, 1974). At a given field
area, a higher leaf population results in a lower nitrogen and alkaloid
concentration such as shown in Tables 1.11 and 1.12. However, for
reducing sugars, a trend for increased levels as leaf populations per
field area were increased has been reported (Table 1.13).
Table 1.11 Effect of leaf
population/acre on total nitrogen
concentration.
Leaves per plant
12 16 20 Average
Plants per ha % Total nitrogen
12 000 2.16 2.12 1.86 2.05
16 000 2.06 1.82 1.82 1.90
20 000 1.96 2.11 1.75 1.94
Average 2.06 2.02 1.81

When the condition is changed within the limits found in normal,

commercial production by maintaining a constant total leaf number
(300 000 leaves per ha), the concentrations of nitrogen, alkaloid and
reducing sugar remained at the same level (Table 1.14) despite
variations in row width, plant population and

 Page 13
Table 1.12 Effect of leaf
population/acre on total alkaloid
concentration.
Leaves per plant
12 16 20 Average
Plants per ha % Total alkaloid
12 000 3.10 2.59 1.88 2.51
16 000 2.66 2.19 1.72 2.19
20 000 2.38 2.16 1.60 2.04
Average 2.71 2.30 1.72

Table 1.13 Effect of leaf

population/acre on reducing sugar
concentration.
Leaves per plant
12 16 20 Average
Plants per ha % Reducing sugar
12 000 16.2 18.4 21.0 18.6
16 000 17.9 21.5 21.0 20.1
20 000 19.9 20.0 22.6 20.8
Average 18.0 20.0 21.6

Table 1.14 Total nitrogen, total alkaloids,

reducing sugars at 296 000 leaves/ha.
Plants per ha × Leaves per plant
Row 24 20 1646814114Average
width 700 000
(cm) × 12 × 15 × 18 × 21
% Total nitrogen
107 2.12 2.10 2.12 2.11 2.11
122 2.11 2.10 2.08 2.11 2.10
137 2.12 2.06 2.08 2.10 2.09
Average 2.12 2.09 2.09 2.11
% Total alkaloid
107 2.57 2.47 2.40 2.40
 2.16
122 2.62 2.45 2.37 2.18 2.41
137 2.61 2.42 2.33 2.16 2.38
Average 2.60 2.45 2.37 2.1
% Reducing sugar
107 1.57 14.7 15.8 14.1 15.1
122 16.9 15.7 14.3 14.4 15.3
137 15.6 14.5 14.7 14.5 1.48
Average 16.1 15.0 14.9 14.3

leaf number per plant. Other results indicate that a greater number of
leaves per plant results in lower nitrogen and alkaloid levels (Tables
1.11 and 1.12). Some of the leaf chemistries may be similar, but are
the physical properties and usabilities altered? At least two reports
(Neas, et al., 1978a; Neas, et al., 1978b) suggest no effect on these
two aspects.
c
Fertilization
Fertilization may be the most important human controllable factor
other than time of harvesting for producing good tobacco. Tobacco
farmers control the form, rate, time and method of application of
fertilizers, and each variable is important.
Plant nutrition will be discussed in detail in later chapters; however, a
few comments will be included in this chapter. Only with carefully
controlled fertilizations, particularly of nitrogen, can good quality,
usable leaf be produced. In a comparison of fertilizer rates in which
several critical leaf components were measured in four plant positions
in flue-cured tobacco, the 1680/ha rate produced more desirable leaf
tobacco so far as those major chemical components are concerned for
the particular conditions, as shown in Table 1.15 (Rogers & Mitchem,
1976).
Potassium is another major essential element which affects the
growth, quality and usability of tobacco. A study on the effect of
potassium on leaf and smoke characteristics of flue-cured tobacco
grown on low potassium sandy loam soil (Chaplin, 1980; Tso, 1990)
used a wide range of potassium oxide (K2O) rates from 0 to 270
kg/ha. It is of interest to note that extremely high rates of K2O did not
affect yield and grade, but did have the tendency to reduce tar and
nicotine delivery, as shown in Tables 1.16 and 1.17.
d
Culture
Every step of field cultural practice is important. For example, stage
of topping and subsequent suckering greatly influences chemical
composition. Early and lower topping results in higher total leaf
alkaloids, total N, and in more tar (TPM) and alkaloid in smoke
(Chaplin, 1980), as shown in Table 1.18. The effects of topping are
optimized by controlling suckers by hand and/or chemicals as shown
in Table 1.19. Chemical control using maleic hydrazide (MH)
generally increases leaf sugars more than hand suckering, whereas
manual control favors alkaloid production more. This result is unique
to MH. Other suckercides do not normally affect chemical

 Page 14
Table 1.15 Effect of fertilizer rates on flue-cured leaf chemistry by plant
positions.
Treatment Plant Total Amino Nicotime Reducing WSA1 pH
position nitrogen nitrogen (%) (%) sugars (%)
(%)
1 1.52 0.139 1.21 16.2 3.42 5.28
1125 2 1.57 0.116 1.63 25.1 3.57 5.31
kg/ha
3 1.79 0.129 2.12 21.8 4.46 5.17
4 2.13 0.209 3.29 17.6 4.76 5.04
Average 1.75 0.148 2.06 20.2 4.05 5.20
1 1.62 0.193 1.36 13.7 3.57 5.33
1688 2 1.82 0.177 1.90 16.5 3.80 5.33
kg/ha
3 1.86 0.128 2.19 19.0 4.39 5.21
4 2.23 0.228 3.08 15.6 5.01 5.07
Average 1.88 0.181 2.13 16.2 4.19 5.24
1 2.01 0.246 1.70 7.2 3.32 5.48
2250 2 2.00 0.219 1.99 10.7 3.80 5.37
kg/ha
3 2.00 0.169 2.48 16.6 4.71 5.18
4 2.40 0.226 3.33 13.9 5.22 5.02
Average 2.10 0.215 2.38 12.1 4.26 5.27
1 ml of 0.1 N NaOH to neutralize 1 g of tobacco.

Table 1.16 Effect of potassium fertilization on leaf characteristics of flue-cured

tobacco grown on low potassium, sandy loam soil.
Rate of Yield Grade Nitrogen Potassium Total Reducing Ash
K2O (kg/ha) index (%) (%) alkaloids sugars (%) (%)
(kg/ha) (%)
0 2757 51.8 1.85 1.10 2.42 15.0 9.8
37.5 2925 5.18 1.81 1.41 2.33 14.7 9.5
75.0 3004 53.5 1.62 1.52 2.21 16.3 9.0
112.5 3005 52.5 1.77 1.74 2.35 15.5 9.7
 150 2953 51.7 1.75 1.95 2.41 15.4 10.2
300 2912 50.3 1.78 2.39 2.34 14.8 10.0
LSD (0.05) NS NS 0.11 0.45 NS NS NS

Table 1.17 Effect of potassium fertilization on smoke characteristics of

flue-cured tobacco grown on low potassium, sandy loam soil.
Rate of Filling Tar Nicotine RD* EMC* Count Burn
K2O capacity (mg/cig) (mg/cig) (mm (%) (No/Cig) (min/40
(kg/ha) (g/63.5 mm) H2O) mm)
0 0.96 22.7 2.30 132 13.2 13.7 11.0
37.5 0.93 21.8 2.20 135 13.0 12.5 10.3
75.0 0.97 22.8 2.17 137 13.0 12.6 10.5
112.5 0.95 21.4 2.13 137 13.0 12.2 9.8
150 0.92 20.0 2.07 127 13.0 11.5 9.2
300 0.98 18.9 1.94 147 12.8 11.7 10.0
LSD NS NS 0.20 NS NS 1.2 0.8
(0.05)
*RD: resistance to draw; EMC: equilibrium moisture content.

 Page 15
Table 1.18 The effect of stage of topping on chemical composition and
smoke characteristics of flue-cured tobacco 3-year averages.1
Stage of Total Total Reducing Petroleum Total Total
topping alkaloids nitrogen sugar ether extract particulate alkaloids3
(%) (%) (%) (%) matter2 (mg/g)
(mg/g)
Pre-bud 3.00 1.80 22.5 5.20 53.9 3.96
Bud 3.15 1.80 22.8 5.16 50.1 3.96
Normal 2.71 1.76 22.4 5.35 51.2 3.60
Late 2.61 1.72 22.5 5.27 49.9 3.29
No 1.70 1.73 20.4 5.70 49.9 2.29
topping
LSD 0.18 0.06 1.5 0.30 3.1 0.22
(0.05)
(0.01) 0.24 NS NS 0.40 NS 0.29
1Combined data from fourth and fifth primings.
2 Total particulate matter on Cambridge filter.
3 Total alkaloids on Cambridge filter.

Table 1.19 Effect of maleic hydrazide

(MH) on reducing sugar and total
nitrogen.
Reducing Total nitrogen
sugar
Topped Manual MH Manual MH
(%) (%) (%) (%)
Button 19.2 23.5 2.36 2.04
stage
Early 20.3 23.9 2.24 2.08
flower
Full 19.4 21.2 2.27 2.28
flower
Late 17.1 18.4 2.36 2.29
flower
composition differentially (Tso, 1990). The residues of MH on
tobacco have been an issue for many years due to the establishment of
MH residue tolerances in some countries. Several studies emphasize
that proper agronomic practices can reduce the residues (Davis, et al.,
1974; Sheets & Leidy, 198796). Also, weather conditions influence
the residue levels. Under droughty conditions, the residues tend to be
higher.
One question frequently raised is why flue-cured and burley types are
so different. Are those differences due to variety (genetic background)
or to culture (environment and human intervention)? One approach to
the answer was provided by a joint study at Oxford, North Carolina, a
typical flue-cured area, and Lexington, Kentucky, a typical burley area
(Chaplin, 1980). Each location follows their own cultural practices
and curing (Table 1.20). It is evident that genetic background,
environment and human intervention are all involved.
e
Maturity and Harvesting
Next to fertilization, time of harvesting is probably the other most
important crop management practice in producing good tobacco.
Maturity generally describes the state of maximum accumulation of
dry matter. Physiologically, when the amount of dry matter reaches
the peak in the life of a plant, breakdown begins to exceed dry matter
accumulation and senescence is initiated. The practical term 'ripe'
usually represents the stage of early senescence. The changes during
senescence continue and intensify during leaf curing. However, if a
leaf is harvested prematurely before senescence has proceeded
sufficiently that leaf cannot be cured to the desired 'quality'
characteristics to attain highest usability. From a biochemical sense, it
is better to harvest leaf 'over mature' than 'prematurely'. Table 1.21
shows chemical composition at visually immature, mature and over-
mature stages. Leaves harvested ripe, i.e. at visual maturity, contain
lower concentrations of nitrogenous fractions and higher nicotine than
the physiologically mature stage (Terrill, 1974). A comprehensive
report on the interactions between maturity and curing was published
by Weybrew, et. al. (1984). The general phenomenon is that the
nitrogenous components of green leaves decrease with delay in
harvest date, also shown by a separate study (Table 1.22).
f
Curing
Curing is another critical factor which decides leaf quality and
usability. In 1996, Peele, et al. presented a summary of the chemical
and biochemical changes

Table 1.20 Comparison of plastid pigment and polyphenol concentrations in tobacco chloroph
under two cultural practices, averaged from samples taken during the growing season and afte
Plastid pigments
ChlorophyllChlorophyllChlorophyll Total CarotenoidsChlorogenic
genotype a b chlorophylls acid
mg/g dry weight
Practices NC 95 2.18 0.67 2.85 0.31 34.71
for flue- NC 95-Py 1.83 0.54 2.37 0.28 29.17
cured SC 58 2.06 0.64 2.70 0.31 35.12
tobacco
SC 58-yg 1.31 0.37 1.68 0.22 32.77
Practices NC 95 3.32 0.90 4.22 0.50 21.25
for NC 95-Py 2.58 0.68 3.26 0.40 18.69
burley SC 58 3.00 0.81 3.81 0.47 25.95
tobacco
SC 58-yg 1.95 0.43 2.38 0.32 25.37
Burley 21 2.21 0.58 2.79 0.38 12.83

 Page 17
Table 1.21 Effects of maturity at harvest on cured leaf composition.
Harvest Total Protein a-Amino NicotineSugar Starch as
treatment N N N glucose
Composition (%)
Immature 2.68 1.10 0.176 2.60 20.8 2.77
Mature 2.49 1.04 0.144 2.75 20.4 2.58
Over-mature 2.44 0.98 0.124 2.95 18.5 2.54

Table 1.22 Sampling date and the nitrogenous fractions of lamina.

Days after TotalInsoluble Soluble a- Nitrate AlkaloidAmino
transplanting N N N Amino N N Acid
N N
(Mg N/100 g fresh weight)
Lower leaves
45 449 362 87 1.52 13.6 19.2 13
53 291 209 82 1.22 13.1 17.6 11
63 199 146 53 0.63 3.7 16.7 4
Upper leaves
45 1050 865 185 12.26 10.1 6.5 31
79 412 291 121 1.24 0.1 63.1 9
99 428 248 180 0.44 0.1 116.0 6

during flue-curing. Wiernik, et al. (1996) discussed the effect of air-

curing on tobacco chemistry. The first stage of flue-curing is a
physiological process during which the biochemistry of harvested
leaves is changed in a desired direction under full or partial control of
temperature and relative humidity, depending on the type of tobacco.
This part of the process, commonly referred to as 'yellowing', requires
the leaves to remain alive. It is terminated by drying, referred to as
'fixing color'. The process ends with practically complete dehydration
to ensure preservation of the leaves for subsequent processing, storage
and manufacture. The control of these processes is greatest in flue-
curing, more so than in air-and sun-curing, which depend largely on
natural variations in temperature and humidity.
Although starch hydrolysis is the principal biochemical activity in
curing, other compounds also are broken down. Additionally, some
recombination compounds are produced and new ones generated, as
shown in Tables 1.23 and 1.24. The result is the characteristic color,
taste and flavor of each tobacco type.
The two major curing methods, flue-curing (bright tobacco) and air-
curing (burley, Maryland and cigar tobaccos), produce different
results, even using the same cultivar (Chaplin, 1980). This is
demonstrated in
Table 1.23 Changes in composition of tobacco
during the flue-curing process.
Constituents GreenYellowed Cured
% Dry weight
Starch 29.30 12.40 5.52
Free reducing sugars 6.68 15.92 16.47
Levulose 2.87 7.06 7.06
Sucrose 1.73 5.22 7.30
Crude fiber 7.28 7.16 7.34
Total nitrogen 1.08 1.04 1.05
Protein nitrogen 0.65 0.56 0.51
Nicotine 1.10 1.02 0.97
Ash 9.23 9.24 9.25
Calcium 1.37 1.37 1.37
Oxalic acid 0.96 0.92 0.85
Citric acid 0.40 0.37 0.38
Malic acid 8.62 9.85 8.73
Resins 7.05 6.53 6.61
Pectinic acid 10.99 10.22 8.48
pH value 5.55 5.64 5.55
Carbonyls (mg/100 g 94.90 610.00 888.00
tobacco)


 Page 18
Table 1.24 The effect of coloring time and
temperature on certain chemical components of flue-
cured tobacco.1
Temperature Time NicotineNornicotine N Reducing
(°C) (hours) (%) (%) (%) sugars
(%)
32.2 48 0.97 0.03 1.36 21.91
72 1.16 0.08 1.53 17.50
37.8 48 1.30 0.02 1.66 17.37
72 1.58 0.07 1.85 14.33
43.3 48 1.36 0.03 1.75 14.32
72 0.81 0.01 1.33 19.60
1 Analyses were performed on composite samples
from five tests.

the study shown in Table 1.25, in which one Maryland cultivar and
two bright cultivars are compared. Leaf and smoke analyses differ
significantly between the cultivars. Each tobacco type is somewhat
different, but in all types leaves are separated according to well
defined visual and physical criteria, which reflect their chemistry and
potential smoking and manufacturing characteristics, to facilitate
selling and subsequent blending.
Even using the same air-curing method, the chemical composition of
leaves changed when primed either prior to curing or cured on the
stalk, such as shown for nitrogenous compounds in cigar tobacco
(Legg & Collins, 1974) (Table 1.26). Another example is the
concentration of nitrosonornicotine (NNN) in air-cured primed leaf,
bulk-cured primed leaf and conventional stalk-cured burley (Chaplin,
1980). Table 1.27 shows the great differences.
During curing, aging or fermentation a 'browning' reaction is known
to take place. Many products are formed which are important to
smoke flavor (Bates, et al., 1974; Leffingwell, 1976). Table 1.28
illustrates a few of those products and their additive nature to smoke
flavor.
An experimental curing method, homogenized leaf curing (HLC)
(Tso, 1972; Chaplin, 1980; Tso, 1990) has been conducted and the
cured material was subsequently reconstituted into sheet using two
different
Table 1.25 Curing methods contributed to chemical changes.
Flue-cured Air-cured
Cultivar MD609 NC2326 Coker 319 MD609 NC2326 Coker 319
(MD) (Bright) (Bright) (MD) (Bright) (Bright)
%
A. Leaf
analysis
Total Alk 2.4 2.5 2.3 0.8 1.1 1.0
Total N 2.3 2.0 2.1 1.8 2.0 2.2
TVB 0.4 0.4 0.4 0.3 0.3 0.3
PEE 5.6 4.7 5.3 6.7 8.4 8.2
Ash 14.2 10.9 11.4 23.3 19.1 20.2
Cl 0.50 0.46 0.50 0.82 0.91 0.89
B. Smoke
analysis
Nicotine
mg/g cig 2.31 2.97 2.80 0.67 1.00 0.89
mg/g tob 2.63 2.72 2.70 0.84 1.22 1.16
burned
Tar
mg/g cig 26.69 32.63 31.00 18.63 23.90 23.31
mg/g tob 30.32 29.94 31.00 23.53 29.15 30.2
burned
Alk: alkaloids; N: nitrogen; TVB: total volatile bases; PEE: petroleum ether
extractables; Cl: chloride; cig: cigarette; tob: tobacco.


 Page 19
Table 1.26 Changes of nitrogenous
compounds in aircured cigar tobacco.
Primed leaf Stalk cured
Types of Before After Before After
nitrogen curing curing curing curing
% Harvested by dry weight
Total 5.61 5.34 4.70 3.80
Protein 3.69 1.65 3.80 1.85
(insoluble)
Soluble 1.92 3.69 0.90 1.95
Amino 0.23 0.80 0.15 0.15
Amino 0.15 1.07 0.05 0.80
plus amide
Alkaloid 0.35 0.32 0.40 0.40
Nitrate 0.63 0.77 0.20 0.25
Reminder 0.56 0.73 0.10 0.35

Table 1.27 Nitrosonornicotine (NNN)

content of aircured primed leaf, bulk-cured
primed leaf and conventional stalk-cured
burley tobacco.
Stalk position
Curing Below top Top leaves
treatment leaves (µg/g) (µg/g)
Stalk cured 1.25 0.79
Primed leaf 1.66 2.17
(aircured)
Primed bulk- 2.50 2.38
cured (leaf)

Table 1.28 Some browning reaction

products of amino acids (or ammonia)
and sugars present in tobacco and/or
smoke.
Products Additive smoke flavor
(number) (individually)
Acids (4) Pungent, buttery, sweet,
Turkish
Aldehydes Pungent, harsh, sweet,
(15) nutty, spicy, fruity
Ketones (12) Sweet, fruity, smoothing
Furans (11) Sweet, herbaceous,
roasted, oily
Pyrans (2) Sweet, flue-cured like
Pyrazines Buttery, nutty, earthy,
(14) burley, dull
Pyrroles (7) Sweet, cheery, hot,
peppery
MiscellaneousSweet, flue-cured like,
(3) burley like

processes. The final products were made into cigarettes and compared
with sheets from both bright and burley tobacco types. Delivery of
nicotine, tar and dimethyl-nitrosamines was greatly modified, as
shown in Tables 1.29 and 1.30.
g
Processing, Aging, and Fermentation
After tobacco is completely cured, it is usually stored on the farm and
prepared for market. The length of this storage varies from a few days
to several months. In some instances the individual farmer's lot may
be combined with others to make a village 'bale'. Each tobacco type is
handled somewhat differently and the terms used to describe the leaf
differ. Even within a type, local tobacco farmers used different
terminology. In one example of classification (for flue-cured tobacco)
the main criteria are color, thickness, length, grain, roughness,
elasticity and the percentage of physical imperfections. Also, leaves
are separated according to stalk position. Bottom leaves, which are
usually thin and smooth, are designated as primings. If they have
some thickness and porosity, they fall into lug grades. Leaves from the
third and fourth priming are normally large and thin and have
considerable grain and oil, and are relatively free from physical
imperfections. They are referred to as cutters. Leaves usually become
thicker above the midstalk. Grades from these upper stalk positions
are known as leaf or, if they are completely ripe, as smoking leaf. All
grade groups have many subdivisions based on color, length,
percentage of mixture with other grades, width of leaf, degree of
physical damage etc. Currently in the United States, and indeed
elsewhere, greater emphasis is being placed on degree of maturity
(ripe, mature, unripe and immature), with less emphasis on separating
qualities within a style (Bowman, et al., 1988).
The procedures for marketing tobacco differ significantly among
countries. They vary from an auction system to direct sales to the
tobacco buyers. This aspect of the chain is the focus of a subsequent
chapter in this monograph.
From sale of leaf to manufacture of product are as important as field
production. The processing of leaf following marketing, but preceding
manufacturing, is important to the development of desirable smoking
attributes. Many consider this requirement to be at least 2 years'
duration. In modern production, purchased leaf is threshed in
specialized processing factories to separate stem from lamina and then
re-dried to a uniform, critical moisture level before packing and
storage.
Freshly cured tobacco leaf is not suitable for use because of its
pungent and irritating smoke. By process of aging and fermentation,
the leaf delivers mild, aromatic smoke. Aging is generally applied to
cigarette tobacco; i.e., a mild state of fermentation, usually carried out
in hogsheads or bales in compressed conditions for several years in a
moisture content ranging


 Page 20
Table 1.29 Nicotine and tar delivery of experimental cigarettes.
Bright tobacco Burley tobacco
Treatment Cig. wt Nic Tar Cig wt Nic Tar
(mg) (mg/cig)(mg/cig) (mg) (mg/cig)(mg/cig)
Conventionally cured leaves 1153 3.77 41.8 989 2.16 24.6
(control)
Slurry sheet of control leaves 1098 1.09 25.1 1150 0.73 17.6
Slurry sheet of homogenized 1071 1.35 28.7 1001 1.24 21.1
leaf curing tobacco
Paper sheet of control leaves 1078 1.27 29.3 1075 0.80 16.1
Paper sheet of homogenized 1054 0.77 22.3 1126 0.63 8.8
leaf curing tobacco
Cig wt = cigarette weight.
Nic = nicotine.

Table 1.30 Nitrosamines in smoke of

cigarettes made from conventional and
homogenized leaf curing.
Samples Dimethylnitrosamines
(ng/g tobacco burned)
Burley tobacco
Control leaves 97.0
(conventionally
cured)
Reconstituted 65.0
sheet of control
tobacco
Reconstituted 20.0
sheet of
homogenized leaf
curing tobacco
Bright tobacco
Control leaves 2.6
(conventionally
cured)
 Reconstituted 1.7
sheet of control
tobacco
Reconstituted <1.7
sheet of
homogenized leaf
curing tobacco

from 1013%. During the process, flue-cured tobacco loses 1 to 2% of

dry weight and air-cured, 3 to 4%. The process, which is the most
active in warmer months, is referred to as sweats (not as severe as the
fermentation described for cigar tobaccos). Fermentation is a standard
practice in cigar tobacco; often here it is referred to as sweating due to
its severity. It is characterized by high initial moisture content (may
reach 50%), by heat generation and by l0 to 20% loss of dry weight.
In addition to the improvement of leaf aroma, there is a change to a
uniform and darker color and a grainy leaf texture. During the
fermentation process there is an evolution of carbon dioxide, ammonia
and other nitrogenous compounds and production of methyl alcohol;
uptake of oxygen; change of pH; change in water retention; and the
improvement of fire-holding capacity (Tso, 1972).
Aging or fermentation brings leaf to its best quality and usability. It is
a joint function of biochemical, biological and physical processes.
h
Reconstitution and Expansion
Reconstitution and expansion are now common practices in the
tobacco industry. In the early 1950s, reconstitution was practised first
in cigar tobacco to produce a product referred to as homogenized
tobacco. The reconstitution process was developed gradually and used
on cigarette tobacco. It makes use of dust, stems and other byproducts
of processing and manufacturing, although straight tobacco materials
are also sometimes used. Reconstituted sheet may be cut into any size
shreds. The sheet making process includes dust improvement,
slurring, impregnation-of-web, paper making process and extrusion
(DeBardeleben, 1980). The exact reconstitution process varies among
manufacturers as demonstrated by the patent literature.
Expansion is a process which increases the shred filling power. One
form is so-called puffed tobacco, whose particle size has been
increased by a process that combines heat, high pressure differential
and a puffing agent to expand the tobacco. In another process, freeze
drying, the cut rag is first wet, then frozen and finally dried in a
vacuum chamber (DeBardeleben, 1980).
One experimental curing method, homogenized leaf curing (HLC)
(Tso, 1972; Chaplin, 1980) that produced tobacco material that was
subsequently reconstituted into sheet was tested in both flue-cured and
burley tobacco types. Delivery of nicotine, tar and

 Page 21
dimethylnitrosamines is greatly modified, as shown in Tables 1.29 and
1.30.
i
Blending
Blending is the selection and thorough mixing of the tobacco-based
components plus any associated casings, humectants and flavorings
required for a particular product or brand specification. The tobacco-based
components may include leaf lamina, cut and rolled stem, reconstituted
sheet and expanded tobaccos. Blenders select individual components for a
blend on the basis that each tobacco type, source (including growing
conditions in the particular year), style and quality has characteristic
physical and chemical properties that influence manufacturing and smoke
parameters. Because the crop is strongly influenced by growing conditions,
especially weather, tobacco is a variable raw material. To ensure that
delivery by a particular blend is consistent, much of the blender's skills are
therefore directed towards minimizing the effects of this variability by
selectively using specific grades and sources of leaf demanded by the
changing situation (Tso & Chaplin, 1977; Tso, et al., 1983).
Although current manufacturing technology can design or manipulate the
input parameters (tobacco, filters, paper, etc.) of a cigarette to meet the
market needs, it is most desirable to start with basic leaf materials of good
quality and usability.
In the United States, plant breeders meet certain standards for new cultivar
releases. Such standards are beneficial for both farmers and industry to
ensure maintenance of quality and usability. The reason for such standards
includes the occurrence of wide chemical differences such as nicotine,
variations in agronomic and disease resistance and resulting smoke
properties of some cultivars (Matzinger, et al., 1984) as shown in Table
1.31, which are the results of genetics, environment and agronomic
practices used by growers.
The principal tobacco types used for cigarette manufacture are flue-cured,
air-cured burley, Oriental and Maryland. Each one plays a different role in
formulating the final product. For example, Table 1.32 shows what
constituents flue-cured and burley contribute to the smoke (Gurin, et al.
1977). Among other things, flue-cured imparts taste, flavor and acidic
aspects; burley provides impact; Maryland influences burn characteristics;
and Oriental contributes to the aroma.
Using bright (flue-cured) tobacco as a relative standard, calculations have
been made (Gurin, 1976; Griest & Guerin, 1977; Gurin, et al., 1977) to
determine the quantity of smoke constituent per cigarette from burley,
Oriental and Maryland types as shown in Table 1.33. These data reflect the
basic characteristics of each tobacco type and, more importantly, the art of
blending as a controlling factor of smoke constituents.
Levels of volatile acids differ among tobacco types and cultivars (Rogers &
Mitchem, 1976). Table 1.34 shows the amount found in two bright (flue-
cured) cultivars (aromatic A and aroma-deficient A) in comparison with
one Oriental tobacco, Samsun.
j
Manufacturing
Cigarette manufacturing involves developing a specific tobacco blend
utilizing the desired tobacco types, breaking up the tobaccos stored in
hogsheads or bales, combining the blend components, cutting the raw
tobaccos into specific dimensions, applying casing and top dressing
materials (unless it is a no-additive product), adjusting moisture content,
selecting the
Table 1.31 Cultivar differences of flue-cured tobacco for nicotine concentrations and
smoke properties.
Nicotine
CultivarsTobacco Smoke Smoke WTPM/NicotineNumber WTPM WTPM Weight
(%) (mg/cgt)(mg/puff) (mg/cgt) of puffs (mg/cgt)(mg/puff) (g/cgt)
Coker 1.5 1.3 0.16 20.8 8.0 27 3.30.97
129
129
PD 33 1.6 1.4 0.17 20.0 8.3 28 3.4
Hicks 2.7 2.5 0.31 13.2 8.0 33 4.1
NC 402 2.9 2.3 0.29 12.6 7.0 29 3.7
NC 95 2.9 2.5 0.32 12.0 7.7 30 4.0
SC 85 3.4 3.0 0.37 11.3 8.0 34 4.3
WTPM = wet total particulate matter (water + dry particulate matter of smoke
condensate collected on a Cambridge filter pad of a smoking machine).

 Page 22
Table 1.32 Constituents of bright and burley tobaccos relative to cigarette smoke
composition.
Quantity per Quantity per gram of Quantity per liter of
cigarette tobacco burned smoke
Constituent Bright Burley Bright Burley Bright Burley
mg
TPM 36.22 24.92 46.13 36.42 111.2 102.8
Water 3.41 3.27 4.35 4.78 10.53 13.48
Nicotine 1.66 1.56 2.11 2.27 5.13 6.40
Tar 31.16 19.65 39.71 28.70 95.85 81.12
CO 18.67 17.88 23.81 26.22 59.27 78.59
CO2 54.27 45.17 69.21 65.23 172.3 198.5
µg
NO 78.5 870.4 99.5 1269.0 252.2 3937.0
HCN 393.7 238.4 503.1 350.7 1186.0 1038.0
Acrolein 82.7 80.5 105.8 119.0 253.7 345.4
Acetaldehyde 797.3 730.7 1020.0 1079.0 2453.0 3166.0
Formaldehyde 26.8 25.1 34.1 36.8 79.2 111.3
Isoprene 750.0 335.0 960.0 495.0 2325.0 1450.0
TPM phenols 163.3 84.25 208.8 123.3 522.8 351.0

appropriate filter, paper, cigarette design properties, combining these

components into a rod (usually on a high speed machine), inserting
the rods into a pack, placing the packs into cartons and consolidating
the cartons into cases for distribution. This is a complex set of steps
which varies among brands. Many of these steps are used in cigar
production and, again, are brand-specific. Smokeless tobacco
manufacturing involves selecting tobaccos for the blend, cutting these
into the desired particle size, applying the selected casings and top
dressings, adjusting moisture content to the level needed for the
particular product, placing the product into containers and
consolidating the containers into cases. Each of these will be covered
in greater detail in the specific chapters of this monograph.
The terms 'flavor' and 'aroma' of tobacco are used rather loosely.
Generally, flavor refers to the blend of taste and smell sensations
evoked by a substance in the mouth; aroma refers to the distinctive
pleasing aroma derived from either leaf tobacco or smoke
(Schumacher, 1984). Additives or neutralizers may be used during
manufacturing to enhance the desirable or reduce undesirable aromas,
respectively.
Roasting of tobacco improves flavor and aroma (Leffingwell, 1976).
Table 1.35 shows the decrease of several free amino acids after
roasting of the complete cigarette blend. In another study (Table 1.36)
shredded samples of flue-cured, Oriental, burley and reconstituted leaf
were roasted for 4 hours at 120 ± 3°C, with an over 2000-fold
increase in dimethylpyrazine concentration being observed in the
latter two types of tobaccos (Leffingwell, 1976).
Smoke
a
Smoke and Tobacco Constituents
Smoke is a complex body of pyrolytic products which varies with
many factors, including raw materials, design of the smoking articles,
manufacture, combustion environments and the way in which the
product is smoked. Smoking is not aimed at complete combustion of
the smoking materials.
Reports on new discoveries of tobacco and smoke components are
published almost every day. Any report of total numbers of
compounds found in tobacco and/or smoke is therefore subject to
question because of the rapid change. Approximately 5000
compounds have been identified in smoke and 4000 in tobacco, many
of which are common to both (Dube & Green, 1982; Green &
Rodgman, 1996). Another report (Roberts, 1988) separated
compounds into functional groups that have been identified, as shown
in Chapters 8A and 8D of this monograph.

 Page 23
Table 1.33 Calculated quantities of smoke constituents per cigarette
relative to those from flue-cured tobacco.
Constituents Tobacco
type
Burley Oriental Maryland
Acidic compounds
Formic acids 0.52
0.48
Acetic acid 0.54
0.45
Propionic acid 0.03 0.66
0.56
t-Butyric acid 0.63
v-Butyric acid 0.94
t-Valeric acid 0.50
Total steam volatile acids 0.59
Total C4-C7 0.61 1.8 0.72
Palmitic acid 0.06 0.60
Phenol 0.44 0.55 0.79
0.32 0.73
o, µ, p-Cresols 0.59 0.91 0.92
0.18 0.61
Catechol 0.93 0.84
Basic compounds
Ammonia 15.6
2.8
Nicotine 0.73 0.29
1.29 0.94 0.83
Alkaloids except nicotine 1.11 0.35 0.36
Volatile bases 1.30 1.43 1.05
Hydrocarbons
C12C33 Paraffins 0.73 1.7 1.2
Benzo (a) pyrene 0.35 0.83
 Benzo (a) pyrene + benzo (e) pyrene 0.57
Benz (a) anthracene + chrysene + 0.51
triphenylene
Pyrene 0.43
Flouranthene 0.58
Phenanthrene + anthracene 0.48
Total volatile neutrals 0.59 0.71 0.50
Isoprene 0.83 0.45
Carbonyl compounds
Formaldehyde 0.53
0.52 0.48
0.84
Acetaldehyde 1.1
1.4 0.71
Propionaldehyde 0.89 0.86
1.1
Acrolein 1.1 0.85
0.73 0.67
Acetone 1.4 1.1
Methyl ethyl ketone 0.71
0.78 0.94
Diethyl ketone 1.1
Inorganic gases
Carbon monoxide 0.95 0.79 0.76
Carbon dioxide 0.72
Nitric oxide 5.2
16.0
7.8 1.1
Hydrogen cyanide 1.0 1.0

 Page 24
Table 1.34 Levels of volatile acids in various
tobaccos.1
AromaticAromadeficient Turkish
Acid A A (Samsun)
Bright
Formic acid 2303 1773 852
Propionic 209 148 153
Isobutyric 26 5 12
Butyric 38 29 45
Isovaleric 181 62 170
Valeric 70 44 29
b- 211 87 1021
Methylvaleric
n-Caproic 67 38 82
n-Heptylic 119 70 82
n-Caprylic 34 44 82
n-Pelargonic 46 180 99
Unknown2 139 105 149
Total 3442 2585 2776
1 Moisture-free basis, µg/100 g tobacco.
2 Calculated as n-caproic acid.

Table 1.35 Change in amino acid

levels in tobacco after roasting
(complete cigarette blend).
Before After Change
(%) (%) (%)
Aspartic 1.39 0.82 -42
acid
Proline 0.65 0.31 -52
Lysine 0.32 0.10 -69
Histidine 0.24 0.16 -33
Arginine 0.16 0.06 -62
Table 1.36 Effects of roasting on
dimethylpyrazine concentration.
Concn prior to Concn after
roasting (ppb) roasting (ppb)
Flue-cured or 0.1 0.3
Bright
Turkish 0.1 0.3
Burley 0.1 460.0
Reconstituted 0.1 200.0
leaf
Tobacco samples were shredded, without
additives and roasted for 4 hours at 120 ± 3°C.

During combustion, various leaf components serve as precursors to

different components in tobacco smoke through pyrolysis. Most of the
pyrolytic products obtained under laboratory conditions are found in
tobacco smoke (Kawashima & Gamon, 1980), as shown in Table 1.37.
Even more dramatic results were obtained in a comparable
observation of major aromatic acids in the smoke from Oriental
tobaccos (Table 1.38) (Tso, 1990).
Table 1.37 Contribution of pyrolysis
products to tobacco smoke.
No of Tobacco
pyrolysis smoke
products/ constituents
Leaf precursors category
Carbohydrates 141 115
(mono-and
disaccharides,
cellulose, starch)
Pectic substances 6 6
Amino acids and 159 122
proteins
Nicotine 58 51
Tobacco leaf pigment 19 19
Reaction of sugars 353 138
with amino acids and
ammonia

Table 1.38 Major aromatic acids in Oriental

tobacco smoke.
Acid Tobacco Smoke %
(µg/g) (µg/cigarette) Change
Propionic 17.0 66.8 +293
Butyri 2.5 9.7 +288
Valeric 4.3 3.6 -16
2- 2.1 12.4 +490
Methylbutyric
Isovaleric 5.1 19.7 +286
3- 12.0 62.0 +417
Methylvaleric
Hexanoic 7.5 3.9 -48
Octanoic 1.7 6.2 +265
Phenylacetic 48.9 19.9 -59

In 1997, Hoffmann and Hoffmann published an excellent invited

review of the changing cigarette focus on the period from 1950 to
1995. These authors described changes in cigarette design, including
filters, ventilation, additives, blend components and physical
parameters. The impact of these changes on smoke composition and
yield was discussed.

 Page 25
b
Correlations among Leaf and Smoke Constituents
Through extensive and systematic studies, much knowledge has been
generated on the relationship between leaf characteristics and smoke
constituents (Tso & Chaplin, 1977; Tso, et al., 1983). Smoke delivery
and smoke composition depend on the characteristics of the leaf
tobacco. The effects of genetic and stalk position differences are
reflected in botanical, physical and chemical properties of leaf
tobacco, which in turn are clearly reflected in the smoke constituents
of these experimental samples. These results agree with those of other
parallel studies using selected leaf 'markers' for identification of leaf
quality and usability.
In addition to identifying leaf and smoke constituents, there have been
numerous studies on the relationships of precursors (leaf constituents)
to products (smoke constituents), including the transfer rates of
specific compounds. During the combustion process many complex
reactions occur, such as near complete degradation of some leaf
constituents, transfer of a fraction of other constituents and
pyrosynthesis of new compounds.
While most of the research has focused on the contributions of leaf
chemistry to smoke composition, the physical properties of leaf
influence the formation of smoke constituents. Indeed, physical
properties are as important as chemical composition in affecting
smoke constituents. They influence combustibility which is
particularly important in determining the nature and quality of smoke
constituents.
Stalk position is an important indicator of certain physical and
chemical properties. For example, leaves at lower stalk positions (in
comparison with the upper ones) are generally thinner, faster burning
and higher in potassium, cellulose and total organic acids. These
lower position leaves are lower in total sugar, total nitrogen, nitrate
nitrogen, total alkaloids, total volatile bases, total free amino acids,
total volatile secondary amines, total polyphenols, petroleum ether
extracts, dry total particulate matter (TPM), smoke nicotine, smoke
phenol, smoke hydrogen cyanide (HCN), benzo(a)pyrene (BaP) and
carbon monoxide (CO).
Simple correlations among most of these variables are expected in
view of the common nature of certain variables that can be grouped
together. For example, leaf thickness, rate of burn, moisture
equilibrium, ash content and K are all associated with combustibility,
some positively and some negatively. Several nitrogenous
components, such as total N, ammonia N, total alkaloids, total volatile
bases µ-amino N, amino acids and secondary amines, form another
group which generally is negatively associated with leaf burn. Yet,
many other groups may be generalized for their relationship with
smoke or tar delivery, smoke constituents and biological activity.
Generally, factors that promote burn result in smoke with a lower
level of TPM, nicotine, cresols, total volatile phenols, HCN, BaP and
benz(a)anthracene, but a higher level of acetaldehyde, acrolein and
CO.
Simple correlation and multiple regressions among many variables
have been reported (Tso & Chaplin, 1977; Tso, et al., 1983). The
correlations of selected leaf and smoke variables are listed in Tables
1.39 and 1.40 for flue-cured and burley tobaccos, respectively.
From these relationships, multiple regression formulae have been
derived to predict smoke components based on a few selected leaf
variables. Two examples follow:
Flue-cured tobacco
TPM minus nicotine and water (mg/cigarette) = 19.6908-1.500 × ash
(%) + 3.592 × total N (%) + 7.519 × phytosterols (mg/g)
 R2 = 0.9094
Burley tobacco
HCN (µg/cigarette) = 14.9601 + 2.7226 × stalk position + 2.8606 ×
nicotine (%) + 63.1169 × oxalic acid (%)
R2 = 0.7348
c
Toxicity of Certain Leaf and Smoke Agents
Smoking and public health is a controversial issue which has recently
become more intense. Tobacco has been smoked for centuries and
used for possibly millennia (Tso, 1990; Tso, 1996). The idea that
smoke might cause cancer was first proposed in 1795, followed by
suggestions 100 years later that chemical or physical effects of
tobacco dust might cause lung disease. Not until early in the twentieth
century were case control studies reported, mostly on lung cancer.
Numerous studies were conducted on the association with smoking of
four diseases: cancer of the lung, myocardial infarction, peripheral
vascular disease and chronic obstructive lung diseases. However,
others maintain the conclusion that smoking is one of the factors
contributing to these diseases is 'more fairly described as presumptive
than as proved'.
In attempting to review and summarize available knowledge, the
International Agency for Research on

Table 1.39 Correlations of selected leaf and smoke variables of bright type tobacco.

Table 1.40 Correlations among selected leaf and smoke variables of burley type tobacco.

 Page 28
Table 1.41 Tumorigenic agents in tobacco and tobacco smoke.
Compounds In In
processed mainstream
tobacco smoke IARC evaluation of
(per gram) (per cig) evidence of carcinogenicitya
In laboratory In humans
animals
PAHc
Benz[a]anthracene 2070 ng sufficient
Benzo[b]flouranthene 422 ng sufficient
Benzo[j]flouranthene 621 ng sufficient
Benzo[k]flouranthene 612 ng sufficient
Benzo[a]pyrene 0.190 ng 2040 ng sufficient probable
Chrysene 4060 ng sufficient
Dibenz[a, h]anthracene 4 ng sufficient
Dibenzo[a, i]pyrene 1.73.2 ng sufficient
Dibenzo[a, l]pyrene present sufficient
Ideno[1,2.3-c, d]pyrene 420 ng sufficient
5-Methylchrysene 0.6 ng sufficient
Aza-arenes
Quinoline 12 µg
Dibenz[a, h]acridine 0.1 ng sufficient
Dibenz[a, j]acridine 310 ng sufficient
7H-Dibenzo[c, g]carbazole 0.7 ng sufficient
N-Nitrosamines
N-Nitrosodimethylamine NDd-215 0.1180 ng sufficient
ng
N- 313 ng sufficient
Nitrosoethylmethylamine
N-Nitrosodiethylamine NDd-25 ng sufficient
N-Nitrosonornicotine 0.389 µg 0.123.7 µg sufficient
4-(Methylnitrosamino)-l- 0.27 µg 0.0877 µg sufficient
(3-pyridyl)-l-butanone
N-Nitrosoanabasine 0.011.9 µg 0.14-4.6 µg limited
N-Nitrosomorpholine NDd-690 sufficient
 ng
Aromatic amines
2-Toluidine 30200 ng sufficient inadequate
2-Naphthylamine 122 ng sufficient sufficient
4-Aminobiphenyl 25 ng sufficient sufficient
Aldehydesc
Formaldehyde 1.67.4 µg 70100 µgb sufficient
Acetaldehyde 1.47.4 µg 181400 µgb sufficient
Crotonaldehyde 0.22.4 µg 1020 µg
Miscellaneous organic
compounds
Benzene 1248 µg sufficient sufficient
Acrylontrile 3.215 µg sufficient limited
1, 1-Dimethylhydrazine 60147 µg sufficient
2-Nitropropane 0.731.21 µg sufficient
Ethylcarbamate 310375 ng 2038 ng sufficient
Vinyl chloride 116 ng sufficient sufficient
Inorganic compounds
Hydrazine 1451 ng 2443 ng sufficient inadequate
Arsenic 500900 ng 40120 ng inadequate sufficient
Nickel 20006000 0600 ng sufficient limited
ng
Chromium 10002000 470 ng sufficient sufficient
ng
Cadmium 13001600 4162 ng sufficient limited
ng
Lead 810 µg 3585 µg sufficient inadequate
Polonium (210Po) 0.21.2 pCi 0.031.0 pCi sufficient sufficient
aNo designation indicates that an evaluation by IARC has not been carried out.
bPAH = polynuclear aromatic hydrocarbons.
cThe fourth report of the Independent Scientific Committee on Smoking and
Health (1988) published values for the 14 leading British cigarettes in 1986::
(51.4% of the market) of 20105 µg per cigarette (mean 59 µg) for formaldehyde
and 5501150 µg per cigarette (mean 910 µg) for acetyladehyde.
d ND = not detected.


 Page 29
Cancer (IARC) commissioned a group of 28 invited experts in 1985 to
Lyon, France, and completed the monograph Tobacco Smoking as a
separate volume of the IARC series Evaluation of the Carcinogenic
Risk of Chemicals in Humans (World Health Organization, 1986).
This volume contains in-depth conclusions and evaluations. In recent
years, tobacco smoke, including mainstream smoke and
'environmental tobacco smoke' (ETS), has been reported to be
associated with many other health problems. The USA Surgeon
General provides an annual report on various aspects of smoking and
health. The vast amount of findings and reports cannot be fully
discussed in this limited space; current knowledge on toxicity of
compounds reported in tobacco is summarized in Table 1.41 (D.
Hoffmann, personal communication, 1994).
Summary
Tobacco is a controversial but most significant economic crop. It
provides a livelihood for millions of people, billions of dollars in trade
and trillions of dollars in business.
To scientists, tobacco is a most rewarding biological material for
study. Most pioneer research was conducted with tobacco, and most
basic knowledge in plant science was generated from tobacco. A few
significant examples include photoperiodism, genetics and breeding,
growth regulators, pollutants, viruses, plant nutrition, organic
metabolism, photosynthesis and photorespiration, post harvest
physiology and biotechnology. No other plant organism is so well
studied.
This chapter has illustrated the available 1500 tobacco introductions
which possess the wide spectrum of genetic variation. Yet, of the
approximately one million genes in a tobacco plant, only 10 000 are
being expressed at any given time. The determination of the identities
and roles of the other 990 000 may even be more challenging for
scientists.
Prior to 1971, there were approximately 9000 scientific publications
on tobacco; between 1971 to 1989, another 60 000 papers were added.
From 1990 to 1996, at least another 21 000 papers were published.
From the 90 000 scientific reports, not only have we learned about the
basic plant science, but they have contributed to our knowledge of
chemistry, physics, engineering and health science. In the late 1980s,
compounds identified in tobacco and smoke amounted to 3000. Now
the amount is estimated to be well over 5000.
The complex genetic make-up of the tobacco plant, environmental
factors which promote or inhibit the expression of genes and the
numerous steps of human intervention all play a major role in shaping
the final product. Furthermore, when tobacco is used through
combustion, even more variables may be involved in the total
complex of physical, chemical, physiological and biological
phenomena. Scientists may evaluate the simple correlations and
multiple regressions among hundreds of leaf and smoke
characteristics under certain conditions, but it will be difficult indeed
to predict the complications fully when all variables are involved.
This chapter is to introduce the concept that from seed to smoke, each
step is important. Here, in summary, this author also wishes to
emphasize that although much is known about tobacco, even much
more is not yet understood. We have to treat tobacco with respect.
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 Page 32

Chapter 2
Breeding and Genetics
P.D. Legg
University of Kentucky
Lexington, Kentucky, USA

B.W. Smeeton
R.J. Reynolds Tobacco Company
Winston-Salem, North Carolina, USA
Introduction
Most cultivated tobaccos belong to the species Nicotiana tabacum L.
Genetic and cytological studies indicate that this allotetraploid species
originated from a cross between N. sylvestris and one of the species
from the Tomentosae section, most likely N. tomentosiformis. The
long and successful history of tobacco improvement probably started
shortly thereafter. The lack of verifiable records of tobacco growing in
wild habitats indicates that it may have originated in a farmer's field
or was domesticated soon after its origin. Considerable literature
exists on the ceremonial use of tobacco by native peoples of the
Americas and its spread in various parts of the world including the
colonies of North America. However, there is a limited amount of
information on the genetic characteristics of early populations and
how tobacco evolved from an interspecific cross into a highly
diversified species.
The major tobacco classes, namely flue-cured or Virginia, light air-
cured including burley and Maryland, dark air-cured, fire-cured, sun-
cured, Oriental or Turkish, and cigar filler, binder and wrapper existed
prior to the initiation of modern breeding research. Early populations
must have been quite variable probably due to both genetic
segregation following the original interspecific cross and gene
mutations. In addition, growers and users of tobacco must have
actively practised selection as they saved seed for subsequent
planting. This selection by growers in North America and many other
places eventually led to the differentiation of the species into the
classes currently grown. The applications of genetic and other
biological principles to tobacco improvement since the beginning of
the twentieth century have significantly augmented and extended the
advances made by the growers.
In broad terms, a modern breeding project will consist of three phases:
planning, line development and testing. In the planning phase, a
breeder should carefully define the goals to be accomplished.
Generally, this will be correcting a deficiency in existing cultivars or
increasing the desirability of cultivars for one or more traits. Other
aspects of planning include identifying and/or selecting appropriate
germplasm sources, choosing the best breeding method, identifying or
developing techniques needed to evaluate the breeding materials and
to select the best genotypes and outlining tests required to fully
characterize the lines obtained from the breeding effort. The line
development and testing phases are the parts of the project where the
plans are carried out and, hopefully, the objectives accomplished. The
following sections of this chapter provide expanded coverage on these
important aspects of cultivar development.
Breeding Objectives
The ultimate objective of breeding programs is to develop cultivars
with the characteristics desired by growers and manufacturers of
tobacco products. In general, growers are interested in those attributes
that increase profitability such as pest resistance, higher yields,
improved quality and ease of handling and curing. Manufacturers
want leaf with desirable manufacturing properties such as high lamina
to midvein ratios and good filling values, and with the chemical

 Page 33
composition needed to develop consumer products that have
satisfying flavor and aroma and are free from harsh, bitter and
pungent characteristics.
Because new cultivars must meet rather precise requirements for
many physical, agronomic and chemical characteristics, the goal of
most breeding projects is to eliminate or reduce weaknesses in
existing cultivars. The specific trait or traits identified for
improvement will depend upon the characteristics of current cultivars
and germplasm sources that possess the gene or genes needed in a
new cultivar. When all the characters for which improvement is
necessary or desirable exist in two or three genotypes of the same
tobacco class, the objective may be to improve all the traits
simultaneously in one project. If more than two or three germplasm
sources are required, it may be more efficient to attempt the
improvement in two or more steps. When the breeder must resort to
interclass or interspecific crosses, it is usually advisable to focus on
the improvement of one specific trait.
Germplasm Sources
Agricultural personnel, especially plant breeders, realize that
collection and preservation of germplasm are needed to supply the
source populations for future crop improvement. Most organizations
involved in tobacco breeding maintain germplasm collections for their
specific class. A more diverse collection is preserved at the Oxford
Research Station in North Carolina with 125 accessions representing
64 of the 66 wild Nicotiana species and over 2000 different N.
tabacum seed stocks. About 1200 of these, known as tobacco
introductions (TIs), are accessions from outside the USA. The
remainder are domestic cultivars, old varieties and selected breeding
stocks. Many of the accessions have been screened for a wide range of
agronomic characteristics, disease and insect resistances, and
chemical constituents. A duplicate seed sample of this collection is
held for the purpose of long-term storage at the National Seed Storage
Laboratory located in Fort Collins, Colorado (Sisson, 1992b). A
listing of germplasm accessions in many countries was published in
1983 in the Catalogue des Nicotiana which is available from the
Cooperation Center for Scientific Research Relative to Tobacco
(CORESTA), 53, Quai D'orsay, 75340 Paris, France, Cedex 07.
The tobacco introductions and the Nicotiana species have been
especially valuable sources of genes for disease resistance. Most of
the diseases and sources of resistance considered by breeders are
given in Table 2.1. Induced mutations are another source of genetic
variability. However, before initiating a mutation program, the
probability of successfully obtaining favorable mutants should be
carefully compared with expected success from other breeding
methods. First, deleterious changes far outnumber desirable mutants
and second, most mutants are recessive and are undetectable without
extensive progeny testing. Nevertheless, useful mutants in tobacco
have been obtained with irradiation, chemical mutagens, and from
somaclonal and gametoclonal cultures.
The central focus in selecting germplasm is to make sure the parental
materials possess all the attributes to be combined in a new cultivar.
Generally, the most desirable parental genotypes are similar for most
major traits except those specifically related to the objective. In other
words, obtaining a desirable combination of characteristics from
crosses among lines of the same class of tobacco is usually the easiest
to accomplish, followed by a combination of traits across classes
within the N. tabacum species. Utilization of germplasm from wild
Nicotiana species is the most difficult and time consuming. Crossing
barriers and differences in chromosome numbers and homologies are
often encountered in interspecific crosses. In addition, adverse effects
on leaf quality are usually experienced when parental lines have
divergent backgrounds and extensive efforts are required to ameliorate
these effects.
Inheritance of Characters
a
Morphological Traits
Structural properties of tobacco such as leaf shape and size, number of
leaves, plant height, number of suckers and internode length are
important to producers and manufacturers because they influence
handling characteristics, yield and chemical composition. Several
mutants with large effects have been identified for leaf properties,
plant height and internode length (Table 2.2). Genes at two loci are
largely responsible for determining leaf shape (Humphrey, et al.,
1964). In different homozygous combinations, four distinctive leaf
shapes can be recognized, with type 1 being the narrowest and type 4
the broadest. Most flue-cured and dark air-and fire-cured cultivars
have type 2 leaf shapes (Coker 139 and NC 13 are type 3) while
burley and cigar cultivars are generally type 4. Sediyama and
Matzinger (1991) evaluated isogenic flue-cured lines of the four
homozygous leaf shape types and found no significant differences for
days to flower, plant height,

 Page 34
Table 2.1 Sources of resistance to the most destructive diseases of
tobacco.
Source of resistance
Disease Original Modern genotypes
Bacterial wilt TI 448A NC 729, K 149
Black root rot Harrow Velvet, TI KY 14
89 TN 90, KY 907, KY 190
N. debneyi
Black shank Florida 301 NC 60, TN 86, KY 190
N. longiflora L8 hybrids
N. plumbaginifolia Coker 371 Gold
Blue mold N. debneyi, N. PBD 6, Ovens 62
goodspeedii
Chemical mutant
Brown spot Beinhart 1000-1 Banket A1
Cyst nematode N. longiflora Burley 21, VA 81, Kutsaga 10
Fusarium wilt TI 448A Speight G28, KY 14, Jaraiz 1
Mosaic N. glutinosa Coker 176, KY 907, Kutsaga
Mammoth 10
Potyviruses Virgin A Mutante TN 90, KY 907
N. tomentosiformis Breeding line TB 4
McNair 944 NC 602
somaclone
Powdery mildew Kuo-Fan Kutsaga E1
Rootknot N. tomentosa NC 95
TI 1400 SAN and NOD lines
N. repanda, N. RL breeding lines
longiflora
Tomato spotted N. alata Breeding line Polalta
wilt virus
Wildfire N. longiflora KY 907, KY 190
N. rustica WZ hybrids

grade index and reducing sugars. However, yield, number of leaves,

percent alkaloids and percent strip yield were affected by leaf shape.
The mammoth or indeterminate growth habit occurs when a single
gene pair is homozygous recessive (Smith, 1950). Vegetative growth
in these indeterminate types continues until the days become shorter
in late summer. If the growing bud is not removed or damaged, plant
heights of 300 to 400 cm with 40 to 50 leaves are not uncommon. The
non flowering, flue-cured cultivars NC 37 NF and Kutsaga Mammoth
10 are commercially-grown mammoths. In contrast, a short-internode
mutant, also inherited as a monogenic character, has been described
(Legg & Collins, 1982). Burley tobacco plants with this mutant trait
are generally about one-half the height of normal plants and produce
an average of 38 leaves compared with 25 to 28 on normal cultivars
(Legg & Collins, 1982). Commercial utilization of the short-internode
trait is questionable under present harvesting and curing methods.
However, encouraging results have been obtained from research at the
West Kentucky Research Station with dark tobacco lines carrying
genes for both the mammoth and short-internode traits. These lines
produce more than twice the number of leaves of standard cultivars
without much increase in plant height.
Other morphological or physiological characters controlled by
qualitative factors include wrinkled or ruffled leaf surface, petiolate
versus sessile leaf attachment, white flowers versus pink flowers and
parthenocarpy in flue-cured cultivars. This latter phenomenon, which
is particularly evident in cv K 326, is probably governed by dominant
alleles at one locus. Flowers of plants with a recessive gene will
quickly abscise if not fertilized soon after anthesis, but in K 326
empty seed pods will form without fertilization.
The genetics of many other morphological traits have not been studied
extensively for various reasons

 Page 35
Table 2.2 Examples of characteristics known to be controlled by
qualitatively inherited factors.
Characteristic Source of genetic factor Inheritance
Chemical
Alkaloid Recurring mutation Digenic; recessive
conversion
Alkaloid level LA Burley 21, LAFC Digenic; recessive
53 (low)
Duvatriene-ols TI 1223, KY Black Monogenic; recessive
(high)
Duvatriene-diols KDH 960 (high) Monogenic; recessive
Cis-abienol TI 1068 (high) Monogenic; dominant
Sucrose esters KDH 959 (high) Monogenic; dominant
Chlorophyll level
Pale yellow TI 1372, PY 10 Monogenic; dominant
White burley Burley cultivars Digenic; recessive
Disease resistance
Black root rot N. debneyi Monogenic; dominant
Black shank-race N. longiflora Monogenic; dominant
0
Mosaic N. glutinosa Monogenic; dominant
Potyviruses Virgin A Mutante (TI Monogenic; recessive
1406)
Rootknot N. tomentosa Monogenic; dominant
Wildfire N. longiflora, N. rusticaMonogenic; dominant
Morphological
Indeterminate NC 37 NF, KM 10 Monogenic; recessive
growth
Leaf shape Hicks × C 139 Digenic; recessive
Nonglanded TI 1112 Trigenic; partially
trichomes dominant
Nonsecreting TI 1406 Monogenic; recessive
trichomes
Parthenocarpy K 326 Monogenic; dominant
Petiolate leaf Some oriental cultivars Monogenic; dominant
Short internode SI KY 171, SI KY 160 Monogenic; dominant
White flowers VA 509 Monogenic; recessive
Wrinkled leaf Wrinkled C 139 Monogenic; dominant

such as limited differences among cultivars, large amounts of time and

effort needed to collect data, lack of good experimental techniques,
and large cultural and environmental influences. Examples include
stalk size, lodging resistance, angle between leaves and stalk, leaf
thickness and potential number of suckers per plant. However, one
type of dark air-cured tobacco is called one-sucker because the plants
produce relatively few suckers with only one at most leaf axils.
Preliminary observations indicate relatively few genes control the
one-sucker trait.
b
Chlorophyll Level
The most famous and important chlorophyll trait is the white burley
character. The low chlorophyll level in burley is caused by recessive
alleles at two loci. In a study with burley and flue-cured cultivars,
Legg, et al. (1977) developed near isogenic lines for characteristics
other than chlorophyll level. These isogenic lines were significantly
different for yield and 12 of 14 chemical components, indicating
considerable influence of chlorophyll level on agronomic and
chemical traits. Such data do not infer a direct relationship between
genes controlling chlorophyll level and other traits. Rather, the data
are an example showing the importance of the total genotype in the
expression of genes for a given trait.
In addition to the classic burley mutant, several other chlorophyll
deficient mutants have been observed and studied. These mutant
characters are controlled by one or two pairs of qualitatively-inherited
genes. The pale-yellow character, that is characteristic of some South
American air-cured types such as Galpao, has been incorporated into
flue-cured genotypes, but these have not been commercially utilized.
Research in western Kentucky with dark tobacco carrying the pale
yellow trait indicates a potential usefulness in reducing damage from
sunburn and improving the color of cured leaves.

 Page 36
c
Chemical Constituents
Several chemical components including alkaloid content and leaf
surface compounds are controlled by qualitative genetic factors (Table
2.2). Two genetic systems have been identified as influencing the
content of alkaloids produced by tobacco plants. Legg, et al. (1969)
found plants with recessive alleles at two loci produce leaves with
levels of alkaloids around 0.3% on a dry weight basis compared to
more than 3.0% in normal cultivars that have dominant alleles at the
two loci. Standard cultivars were designated AABB and the low
alkaloid lines aabb. Further studies (Legg & Collins, 1971a) showed
the relative contribution of an A gene to be about 2.4 times that of a B
gene. In their study, the average levels of alkaloids were 4.6% for
Burley 21 (AABB), 0.3% for low alkaloid Burley 21 (aabb), 4.2% for
the AAbb genotype and 2.5% for the aaBB genotype. Tobacco
cultivars with the aabb genotype have produced lower yields and
poorer quality leaf than cultivars with other combinations of these
genes.
In addition to differences controlled by genes at the A and B loci,
variation for alkaloid level is present among cultivars that are AABB
at these two loci. Several population studies with both burley and flue-
cured cultivars indicate that these differences are heritable and
controlled by quantitative factors with additive gene action.
Cultivars occasionally include individual plants that contain unusually
high amounts of nornicotine. The ability to convert nicotine to
nornicotine is controlled by two genetic loci (Mann, et al., 1964).
Nonconverting plants, which constitute most of those grown
commercially, are recessive at both loci (genotype c1c1c2c2); whereas,
plants with one or more of these four alleles in the dominant condition
are converters. Mann, et al. (1964) have shown that only the c1 locus
is active in modern cultivars. The locus is unstable and mutates to the
dominant state with high frequency. High levels of nornicotine in the
leaf adversely affect smoking quality and breeders must eliminate
converter plants from their seed stocks. The presence of nornicotine
producing genes in flue-cured tobacco is easily detected by the 'cherry
red' color of the cured leaf. Air-cured tobaccos do not display any
unusual colors and the presence of converter genes can only be
detected by chemical analysis.
Among other chemical components controlled by qualitative factors
are the leaf surface compounds that are synthesized and excreted by
the glandular leaf trichomes. Almost all cultivated tobaccos produce
a-and b-4, 8, 13-duvatriene-1, 3-diols (DVTs). Notable exceptions are
the dark air-cured French cultivars such as PBD 6 which produces
only labdanoids. Concentration of DVTs is influenced by factors
affecting trichome traits and those affecting biosynthesis of DVTs.
The occurrence of glandular trichomes on the leaf surface depends
upon alleles at three loci, with gene effects being partially dominant
(Burk, et al., 1982; Johnson, et al., 1988). Glandular trichomes are
lacking when the partially dominant alleles are present and
homozygous at all three loci, but are present in all other genotypes
with increasing density as the number of partially dominant alleles
decreases. The ability of glandular trichomes to secrete exudate is
controlled by a single, dominant gene (Nielsen, et al., 1982) and the
concentration of DVTs is controlled by a single gene in some lines
and quantitative factors in others (Nielsen & Severson, 1992). Sisson,
et al. (1993) have documented the genetics of duvatriene-ols, showing
them to be conditioned by a single recessive gene pair. Labdanes and
sucrose esters are not present in flue-cured, burley and most dark air-
cured types, but nearly all cigar and Oriental types contain the genes
for their production. They are the precursors of some of the cedary
and valereal flavors associated with the aromatic smoking properties
of these tobaccos. Each constituent is under the control of a different
dominant gene (Tomita, et al., 1980; Gwynn, et al., 1985), but
unpublished work by several breeders indicates that they are linked on
the same chromosome.
Cultivar differences have been found for many other constituents such
as sterols, protein, nitrogenous constituents other than alkaloids,
carbohydrates, polyphenols, carotenoids, solanesol, minerals, organic
acids, fatty acids and lipids. When any of these components has been
studied in segregating populations or cultivar crosses, polygenic
control and additive gene action have been reported.
d
Disease Resistance
Breeders have put forth considerable effort to utilize disease resistance
from N. tabacum genotypes and from those species where crosses
with N. tabacum have been possible. In general, resistances derived
from interspecific hybridization are simply inherited and show
dominance gene action (Table 2.2); whereas those derived from N.
tabacum sources are either controlled by two gene pairs or are
classified as quantitative because scientists have not been able to fit
F2 and backcross data to known Mendelian ratios.
For resistance from N. tabacum sources, genetic

 Page 37
information is often limited to reactions in F1 hybrids and general
behavior in following segregating generations. Studies designed to
obtain inheritance data often fail to yield conclusive results due to the
influence of cultural factors, environmental effects, genotype ×
environmental interaction, homozygous lethality, modifying genes
from recurrent parents, severity and race of disease organisms, and
age and condition of plants at data collection time. In addition,
pathogens are living organisms and thus present another source of
variability not present in experiments designed to measure static traits.
Also, studies designed to measure disease resistance require the use of
precise procedures to prevent contamination by other pathogens or
stress on plants that can change their reaction to pathogens.
Black Shank
The main source of resistance to black shank (Phytophthora
parasitica var nicotianae) in USA cultivars is Florida 301. Prior to the
spread of race 1, the breeding line L8, with a single dominant gene
transferred from N. longiflora (Valleau, et al., 1960), was used to
make commercial hybrids of burley with resistance to the first
identified race of black shank (race 0). The most resistant flue-cured
cultivar is Coker 371 Gold with resistance from Florida 301 and a
single dominant gene which probably came from N. plumbaginifolia
and is the same gene as the one in L8 from N. longiflora. The reaction
of these genotypes varies with the race of black shank present. L8 and
Coker 371 Gold are highly resistant to race 0, while cultivars with
Florida 301 resistance have moderate levels of resistance to both races
0 and 1. Several genetic studies have been conducted with the Florida
301 source of resistance, but conclusive data on the inheritance
pattern have not been reported (Crews, et al., 1964).
Theories about resistance have varied from control by recessive
alleles at one genetic locus to polygenic control with partial
dominance. Several factors, including environmental influences,
probably prevent a satisfactory genetic explanation, but the presence
of modifiers in susceptible lines that enhance or lower resistance has
been cited in many situations. Breeders have made numerous attempts
for many years to utilize the high level of resistance in the cigar line
Beinhart 1000-1, but this resistance has not been successfully
transferred into commercial cultivars. The interaction of genotype
with race is the subject of an ongoing CORESTA collaborative study
(Nielsen, 1993).
Black Root Rot
Resistance to black root rot (Thielaviopsis basicola) from N. tabacum
sources such as cv Harrow Velvet and TI 89 appears to be complex
with some factors being recessive and others dominant. An example
of the complex inheritance pattern was found in crosses involving the
burley cultivars KY 14, Burley 21 and Judy's Pride (Wilkinson, et al.,
1991). Generation mean analyses of parental, F1, F2 and first
backcross generations indicated the presence of additive gene,
dominance gene and epistatic effects. Like the situation with black
shank, Nicotiana species resistance to black root rot has been far less
complicated than that found in N. tabacum sources. Clayton (1969)
successfully transferred resistance from N. debneyi into tobacco and
his crossing experiments clearly demonstrated that resistance was
controlled by a single dominant factor. This single dominant
resistance has been incorporated into most burley and many Canadian
and European flue-cured cultivars released in the past 20 years. Legg,
et al. (1981) developed isogenic burley lines with and without this
resistance. Differences were noted in morphological and chemical
traits and there was a slight reduction in yield. Brandle, et al.
(unpublished data, 1994) have shown that tobacco lines transformed
with the antifungal gene ß-1,3-glucanase expressed high resistance to
this pathogen.
Fusarium Wilt
Resistance to Fusarium wilt (Fusarium oxysporum) is present in USA
cultivars of all types. The original source of resistance in most of
these cultivars is TI 448A. Resistance is inherited in a quantitative
manner (Moore, et al., 1960) and generally provides sufficient
protection against this pathogen. Recent studies in Spain have
identified a local burley cultivar, Jaraiz 1, as being highly resistant and
its resistance is being incorporated into hybrid and standard cultivars.
Blue Room
Most of the resistance to blue mold (Peronospora tabacina) has been
derived by interspecific hybridization of N. tabacum with several
Australian species, including N. debneyi, N. goodspeedii, N. excelsior
and N. velutina. Clayton (1968) found resistance in N. debneyi to be
controlled by several genetic factors and extensive crossing and
selection experiments were required to develop breeding lines with
normal inheritance patterns. Different levels of resistance indicated
the

 Page 38
presence of one, two or three dominant genes from N. debneyi plus a
major factor from tobacco. Breeding work in Bulgaria has confirmed
that resistance from N. debneyi is influenced by up to three dominant
genes (Palakarcheva, 1981). In Australia, resistance transferred from
N. goodspeedii to tobacco was controlled by a single dominant factor
(Wark, 1963). The only other important source of resistance is a
breeding line, Chemical Mutant, which was developed by treating
seeds of a flue-cured cultivar, Virginia Gold, with the mutagen
triethylene iminotriazine. Resistance in this line is thought to be
regulated by a partially dominant gene, but no commercial cultivars
with the resistance have been developed (Rufty, 1989). No USA
cultivars have been released with resistance to blue mold, but many
resistant air-cured and oriental cultivars are widely grown in Europe.
A CORESTA study has recorded the reaction of a set of resistant and
susceptible genotypes in different countries over a period of 30 years
and, although levels of infection have varied over the years, the
resistant genotypes have always provided adequate protection
(Blanchard, 1993).
Powdery Mildew
Cultivars with resistance to powdery mildew (Erisyphe
cichoracearum) are widely grown in Southern Africa (e.g. Kutsaga
E1), Europe and Asia. In most of these cultivars, resistance depends
on duplicate recessive factors originally derived from the cultivar
Kuo-Fan (syn Kokobu)(Wan, 1962). Single dominant genes for
resistance from several species, including N. glutinosa and N.
tomentosiformis, have been transferred into flue-cured breeding lines
(Ternovski, 1941; Palakarcheva, 1981). The reaction of these and
other sources of resistance to this fungus has been evaluated over a
period of 20 years in a CORESTA collaborative study (Fisher, 1993).
The collective results indicate that the Kuo-Fan source of resistance is
the most effective in controlling the fungus.
Alternaria (Brown Spot)
The major source of resistance to Alternaria leaf spot (Alternaria
alternata) is the cigar line Beinhart 1000-1. Breeders in Zimbabwe
have transferred a partially dominant gene from this genotype into
burley and flue-cured cultivars. In flue-cured backgrounds, the
presence of the gene in the homozygous condition causes a delay in
senescence with adverse effects on quality. However, F1 hybrids
between homozygous resistant and homozygous susceptible cultivars
give adequate disease control and acceptable quality. The same gene
in homozygous condition in burley backgrounds (cvs Banket A1 and
Banket 102) has proven to be satisfactory in both respects (Smeeton &
Ternouth, 1992). Several USA flue-cured cultivars, notably K 326,
have useful levels of tolerance to brown spot, presumably through the
action of quantitative factors that breeders have accumulated in the
USA flue-cured gene pool.
Bacterial Wilt
None of the Nicotiana species exhibit resistance to bacterial wilt
(Pseudomonas solanacearum), also called Granville wilt, but
moderate levels of resistance have been found in several N. tabacum
genotypes. TI 448A, with recessive and polygenic resistance factors,
has proven to be the most productive source (Sisson, 1992a). Many of
the newer flue-cured cultivars (e.g. NC 729 and K 149) released in the
USA have resistance from TI 448A. Japanese researchers have
reported high levels of resistance in several domestic types and have
successfully developed resistant burley cultivars from these sources
(Yoshida, et al., 1992).
Wildfire and Angular Leaf Spot
A dominant gene for resistance to wildfire (Pseudomonas syringae pv
tabaci) was introduced into N. tabacum from N. longiflora by Clayton
(1947). Burley 21, the first cultivar to be released with this resistance,
has been used as a parent for crosses in many burley and flue-cured
breeding programs. This gene, while effective against wildfire race 0,
is ineffective against race 1 and only partially effective against the
angular leaf spot (ALS) strain of P. syringae. Breeders in Zimbabwe
have successfully transferred a dominant gene from N. rustica cv
Brasilea into flue-cured tobacco that confers resistance to both races
of wildfire as well as the ALS strain (Woodend & Mudzengerere,
1992).
Tobacco Mosaic Virus (TMV)
A dominant gene conferring hypersensitive resistance to TMV was
introduced into tobacco from N. glutinosa by Holmes (1936). This
gene has been incorporated into most current burley, Maryland and
cigar cultivars and provides complete control of this virus. In flue-
cured types, the presence of the gene generally has adverse effects on
cured-leaf yield and quality and only a few cultivars, such as Coker
176, have this resistance. F1 hybrids of resistant × susceptible
breeding lines produce intermediate leaf yields compared to their

 Page 39
parents, and leaf quality is usually superior to the homozygous TMV-
resistant parent (Wernsman, 1992). Also, the adverse effects
associated with the gene seem to be negligible in mammoth
(nonflowering) flue-cured backgrounds. A high yielding Zimbabwe
cultivar, Kutsaga Mammoth 10, has this resistance and produces
acceptable leaf quality. In contrast to the hypersensitive reaction of
resistance from N. glutinosa, a symptomless resistance is found in TI
1560 (Ambalema) and several other TIs. In Ambalema, resistance is
controlled by two pairs of recessive genes (Clayton, et al., 1938), but
efforts to use these genes in commercial cultivars have failed because
of associations, either via linkage or pleiotropic, with adverse effects
on agronomic traits and quality.
Potyviruses
These aphid-transmitted viruses include potato virus Y (PVY),
tobacco etch virus (TEV) and tobacco vein mottle virus (TVMV).
Resistance is present in a number of Nicotiana species and N.
tabacum genotypes. A cultivar, Virgin A Mutante (VAM), derived by
Koelle (1961) through irradiation-induced mutation, has been used
most often as a source of resistance by breeders. The resistance, which
is controlled by a pair of recessive genes, was first effectively utilized
in the development of the burley cultivar TN 86. The genes provide
resistance or tolerance to most strains of PVY, TEV and TVMV, but
VAM and other natural sources of resistance fail to provide complete
protection against all strains of the potyviruses.
Legg, et al. (1980) found TVMV resistance in Virgin A Mutante is
controlled by a monogenic factor that is recessive in the sense that a
homozygous genotype is necessary to obtain plants completely free
from symptoms. Rating of infected plants indicated that the
heterozygote was intermediate and gene action was completely
additive. Other genotypes with monogenic resistance include Havana
307, Sota 6505E, Burley S-3 and the flue-cured breeding line TB 4
derived from N. tomentosiformis. Allelism tests of these and VAM
indicate that the genes are all allelic and cannot, therefore, be
combined into one genotype to increase levels of resistance
(Wernsman, 1992). A PVY resistant genotype (NC 602) with a
partially dominant gene was identified by Witherspoon, et al. (1991)
in gametoclonal cultures. Since this gene is not allelic to the gene
from VAM, a combination of the two genes in one genotype might
increase the range of potyvirus resistance.
Tomato Spotted Wilt Virus (TSWV)
Work in Poland has shown several Nicotiana species, including N.
alata, to be resistant to TSWV. This species was hybridized with N.
tabacum and resistance stabilized in a genotype called Polalta
(Berbec, 1994, personal communication). Nielsen (unpublished data,
1993) has investigated the inheritance of this resistance and concluded
that two gene pairs may be involved. No other natural sources of
resistance have been reported, but encouraging results have been
obtained with tobacco plants genetically engineered to express part of
the viral coat protein of TSWV.
Miscellaneous Virus Diseases
Many other viruses such as cucumber mosaic (CMV), alfalfa mosaic,
tobacco rattle, tobacco ringspot, bushy top and leaf curl infect
tobacco. Some cause severe losses in certain countries, but for most
there has been limited research on identifying sources of resistance.
Taiwanese workers reported the development of cultivar TT 8 with
resistance to (CMV) (Chen, et al., 1987) and resistance to other
viruses has been reported in the Nicotiana species (Lucas, 1975).
e
Insect Resistance
Rootknot Nematodes
Of the rootknot nematodes that infect tobacco, Meloidogyne
incognita, M. javanica, M. arenaria and M. hapla are the most
important. Additionally, different races for each of these species have
been identified which complicates the process of breeding for
resistance. Resistance to M. incognita races 1 and 3 was derived from
TI 706 and incorporated into NC 95, the first rootknot resistant
cultivar to be released. Apparently, this resistance was originally
under polygenic control, but crosses with a hybrid thought to be
allopolyploid N. sylvestris × N. tomentosiformis produced resistance
controlled by a single dominant factor. Later experiments showed this
single factor comes from N. tomentosa or N. tomentosiformis and is
located on the G Chromosome (Slana, et al., 1977; Slana & Stavely,
1981). Plants with this gene show a necrotic reaction when infected
with PVY strain MN (Rufty, et al., 1983). This phenomenon provides
a simple and effective way to screen segregating generations for the
presence or absence of the gene.
Resistance to M. javanica has been developed independently in South
Africa and Zimbabwe by

 Page 40
combining the M. incognita resistance from a cultivar such as NC 95
and a local breeding line R83 (TI 1400), or its derivative RKT 15-1-1,
which have a different, partially dominant gene for resistance
(MacKenzie & Ternouth, 1986; Cornelissen & Van Wyk, 1987).
While both R83 and RKT 15-1-1 have low resistance to M. javanica,
the combination of genes from both parents provides a high level of
protection. Also, the South African lines (SAN and NOD lines) have
resistance to M. incognita races 2 and 4 and limited protection against
other Meloidogyne species. Several Nicotiana species including N.
repanda and N. longiflora have resistance to M. javanica. Flue-cured
breeding lines (RL lines) which possess genes from both N. repanda
and N. longiflora have shown outstanding levels of resistance in
Zimbabwe (Ternouth, et al., 1986). Another promising source of
resistance to M. javanica is the Japanese cultivar Okinawa. Nicotiana
otophora accession La Quinta has shown resistance to race 4 of M.
incognita (Reed & Schneider, 1992). The interrelationship of all these
sources of resistance has not been established. However, the similarity
of low temperature sensitivity of lines resistant to M. incognita and
those resistant to M. javanica suggests allelism of the two resistances
(Rufty, et al., 1983; Kadotani, et al., 1985).
Cyst Nematodes
Cyst nematodes (Globodera spp) cause severe root damage to tobacco
in some areas, notably in the states of Virginia and Connecticut. Work
by Wilkinson (unpublished data, 1993) has confirmed previous reports
that genotypes with the dominant gene for resistance to race 0 of
wildfire are generally also resistant to G. tabacum solanacearum.
These include Burley 21 and the flue-cured cultivars VA 81, PD 4, NC
567, Kutsaga Mammoth 10, and Kutsaga 110.
Foliar Insects
A comprehensive review of sources of resistance to three major
tobacco insects (hornworms, budworms and aphids) was published by
Johnson, et al. (1992). These authors have shown the mechanism of
resistance is, in most cases, determined by the presence or absence
and the concentration of leaf trichome exudates on the surface of
tobacco leaves. These exudates include the duvanes, labdanes and
sucrose esters. The first flue-cured cultivar, CU 263, with a moderate
level of resistance to tobacco budworms (Heliothis virescens) was
released in the USA during 1995. It is not known whether it has
resistance to other species of budworm that are prevalent in other
countries. Genes controlling the presence or absence of leafsurface
exudate are simply inherited, making them relatively easy to
manipulate in a breeding program. Because labdane and sugar ester
compounds are precursors of aromatic compounds in the smoke, it
may not be possible to develop acceptable cultivars of flue-cured,
burley, Maryland and nonaromatic dark aircured tobacco with insect
resistance based on these constituents.
f
Yield, Quality, and Handling Characteristics
Yield and quality are complex terms used to identify the composite
expression of several components. Among the traits influencing leaf
yield are number of leaves, leaf size, leaf thickness, density of cells,
size of midveins, venation, etc. Quality, as considered here, relates to
physical properties of cured leaves such as color, size, body, texture,
elasticity, structure, oil, uniformity and injury tolerance. Yield, quality
and many of the traits influencing yield show a continuous
distribution in segregating populations and thus are described as
quantitative traits controlled by many genes. Statistical procedures are
used to estimate heterosis and genetic parameters such as types of
genetic variances and heritability. For the most part, traits considered
in quantitative genetic studies have been leaf length and width,
number of leaves, plant height, yield and quality. Yield is generally
measured in kg/ha and quality is expressed as a value based on market
price or as an index based on an official grading standard.
The diallel cross has been used to study crosses among cultivars and
lines within most of the different classes (Legg, 1989). Each study has
produced specific results, but overall the data have been comparable
and similar to those presented in Table 2.3 from experiments in
Kentucky with the burley and dark fire-cured classes (Legg, et al.,
1970; Legg, 1991). Mean performance of F1 hybrids has generally
been comparable to midparent values, but individual crosses have
shown heterosis of 15% or more. Rarely have F1 hybrids exceeded
their best parent or the best parental genotype in the test. Estimates of
general combining ability have been statistically different from zero
for most traits in most studies, whereas large estimates of specific
combining ability variance have been infrequent. General combining
ability shows the average performance of a line in hybrid combination
and is a

 Page 41
Table 2.3 Estimates of variances of general (s2g)
and specific combining ability (s2g) and average
heterosis from midparent in diallels of burley and
dark fire-cured tobacco (Legg, et al., 1970; Legg,
1991).
Diallel and trait s2g s2g Heterosis (%)
Burley
Yield, g plant-1 9.8** 22.6** 4.8**
Leaves, no plant- 1.8** 0 1.5
1
Height, cm 12.9** 1.1 2.1**
Dark fire-cured
Yield 425.8** 35.2 3.8*
Leaves 0.12** 0.06 1.3
Height 3.4 6.5 1.8
*, ** Significantly different from zero at the 5%
and 1% levels, respectively.

measure of additive gene effects. Specific combining ability is present

when specific hybrid combinations deviate from performance
expected on the basis of general combining ability and is a function of
dominance and epistasis. A predominance of additive genetic variance
has also been found in random-mated populations and segregating
generations following the cross of two pure genotypes. Heritability
values have been moderate to high when estimated by parentoffspring
regression or on the basis of self, half-sib, and full-sib families.
Results from three studies in Kentucky are given in Table 2.4 as
examples of data obtained from variable breeding populations (Legg
& Collins, 1971b; Legg & Collins, 1971c; Legg & Collins, 1975).
Breeding Methods
Tobacco is one of the classic self-pollinated species because natural
cross-pollination is relatively low, significant levels of depression are
seldom found following inbreeding and limited heterosis is expressed
in hybrids between inbred lines. The predominance of additive genetic
variance in populations and low levels of heterosis in most crosses
indicate that breeders should focus on the development of pure-line
cultivars. However, there are exceptions where hybrids might be
preferred in commerce over pure-line cultivars. The use of burley
hybrids, with L8 as one parent, to control race 0 black shank is a
classic example where hybrids have made a significant contribution.
Also, for populations with large amounts of nonadditive genetic
Table 2.4 Estimates of additive genetic (s2A)
and dominance genetic (s2D) variances and
half-sib heritability in F2 populations of burley
tobacco (Legg & Collins, 1971b; Legg &
Collins, 1971c; Legg & Collins, 1975).
Population and trait s2A s2D Heritability
KY 10 × Burley 21
Yield, g plant-1 100.4* -14.3 0.37
Leaves, no plant-1 1.1** -0.2 0.39
Height, cm 26.8* -4.8 0.36
KY 14 × Ex 42
Yield 64.1* -7.0 0.34
Leaves 0.4* -0.1 0.29
Height 13.3* 3.8 0.22
VA B-29 × KY 12
Yield 79.4* 39.2 0.66
Leaves 3.8* 1.6*** 0.77
Height 41.6* 22.8*** 0.68
* Larger than twice the magnitude of the
standard error.
*** Larger than once, but smaller than twice
the standard error.
variance and rare crosses exhibiting high heterosis, breeders may need
to develop hybrids to maximize genetic improvements.
Many breeding methods, particularly those listed in Table 2.5, have
been used successfully with the species. The traditional methods of
pure-line selection, mass selection and hybridization with segregating
generations handled by pedigree, backcross or recurrent selection
methods have proven successful. With the refinement of tissue culture
techniques, the use of haploids and doubled haploids has found a
useful place in tobacco breeding. Also, recent advances in genetic
engineering technologies offer great potential for increasing the range
of genetic variability through the introduction of genes into tobacco
germplasm from virtually any biological organism.
Selection for improvement in a single character will generally be
maximized by direct selection for the character. However, the breeder
can seldom focus on one characteristic, but must consider a large
range of characteristics that must be molded together into a cultivar. In
other words, the action of specific genes, genetic relationship among
characters and the influence of the environment determines the final
plant phenotype. Genetic relationships of major concern to breeders
are those that limit progress from selection. These include positive
correlations among number of leaves, plant height and yield which are
restrictive because tall plants are undesirable for field and har-

 Page 42
Table 2.5 Most frequently used breeding methods for tobacco improvement.
Method General features and comments
Backcross Useful in improving cultivars for one or a few traits like resistance to a
disease. Allows systematic transfer of a simply inherited trait to a
cultivar with minimal disruption of other traits of the cultivar. Used in
the development of male sterile lines.
Genetic Allows transfer of genes into tobacco from other biological organisms.
engineering Thus, breeders have the potential benefit of many new sources of
genetic variability.
Haploids- Increases efficiency because recessive allales cannot be masked by
doubled heterozygosity. Allows early testing of homozygotes. Eliminates the
haploids normal inbreeding process and thus can save 1 to 3 years in the
development of a new cultivar.
Mass Similar to pure-line selection except new cultivars consist of several
selection pure lines. This method requires less testing than pure-line selection.
Pedigree Useful in combining desirable traits found in two or more cultivars.
Pure-line Useful in identifying superior lines in a genetically variable population.
selection New cultivars consist of a single pure line.
Recurrent Increases the opportunities for genetic recombination and the
selection frequency of favorable alleles. Thus, the amount of improvement from
a given population is maximized.

vesting operations. Another troublesome correlation is the negative

relationship between yield and nicotine.
a
Pure-Line and Mass Selection
Pure-line and mass selection were the methods of choice for the
development of new cultivars prior to the 1930s when hybridization
became necessary to introduce genes for disease resistance into
breeding materials. The many old-line cultivars still maintained in
germplasm collections attest to the success and importance of these
selection procedures in the history of tobacco production. Most cultivars
now in production for flue-cured and burley tobacco have been
developed from hybrid populations. However, producer-selected
developed from hybrid populations. However, producer-selected
cultivars are still important in other classes. For example, a large part of
the dark flue-cured tobacco in the United States is produced with
selections from the Madole cultivar. These include Narrow Leaf
Madole, VII Madole, TR Madole, Jernigan's Madole, Adkin's Madole,
Head's Madole and Certified Madole.
b
Backcross Method
The breeding procedure of choice following hybridization usually
depends upon the parental material. When the objective has been to
obtain disease resistance or some other characteristic from one of the
Nicotiana species or another class of tobacco, the method of choice has
generally been the backcross or some modification of it. This method
allows a systematic transfer of a simply inherited character to a cultivar
with minimal disruption of other properties of the cultivar. While the
objectives are modification of a single trait and complete recovery of all
the other properties of the recurrent parent, the ultimate result is usually
the introduction of some variability into the recurrent parent for
quantitatively-inherited traits. The transfer of genes for resistance to
several diseases through interspecific hybridization followed by
backcrossing attest to the importance and effectiveness of the backcross
method in tobacco breeding.
c
Pedigree Method
When attributes desired in a new cultivar are found in two or more
genotypes within the same class, the preferred breeding procedure to
combine the traits into one cultivar is usually some form of the pedigree
method. A hypothetical example of the pedigree method, as it might be
used in flue-cured tobacco breeding, was presented by Wernsman and
Rufty (1987). The initial step in utilizing the pedigree method is the
development of a variable population, either an F2 from a cross of two
lines or a random-mated population formed by intercrossing three or
lines or a random-mated population formed by intercrossing three or
more lines. In the first segregating generation, the best plants are
selected on the basis of disease resistance and desirable plant
morphology and then selfed. During the next few

 Page 43
generations, selections are made among and within families. As
homozygosity approaches, selections are made among the lines on the
basis of resistance to disease, desirable plant morphology and
diversity of relationship. At this stage, replicated yield and quality
tests are usually started. Deviations from the traditional pedigree
method might involve one or two generations of bulking or
intercrossing sometime during the process or carrying lines through a
portion of the scheme and then crossing them with another genotype
to bring in one or more additional desirable attributes.
d
Doubled Haploid Method
Relatively easy procedures are available for the extraction of haploid
plants and for doubling their chromosome number to produce
homozygous diploids. Some concern has been expressed about vigor
depression in doubled haploids obtained through anther culture.
However, such depression is not as severe in doubled versions of
haploids produced from egg nuclei (Wernsman, et al., 1984). The use
of haploids and doubled haploids has many potential advantages over
conventional procedures in a breeding program (Collins & Legg,
1974). Greater efficiency is realized from selection for traits
controlled by dominant genes, such as race 0 black shank resistance,
because recessive alleles cannot be masked by heterozygosity. For
quantitative traits such as yield, reliable performance testing can be
done using doubled haploids at any stage in the breeding process
because the breeder can fix the genetic system of individual gametes
in a reproducible form. The utilization of doubled haploids eliminates
the normal inbreeding process and thus can save the breeder from 1 to
3 years in cultivar development. At North Carolina State University,
tobacco breeders are using doubled haploid procedures with
vegetative cloning as their primary breeding method (Wernsman,
1990). In their program, haploid seedlings obtained from crosses of
tobacco plants with pollen of N. africana (Burk, et al., 1979) are
screened in the field or greenhouse for disease resistance. Those
haploids with desirable genotypes are vegetatively cloned, and leaf
midvein explants are cultured to produce doubled haploids for
evaluation in replicated field tests.
Doubled haploid breeding requires the breeder to have tissue culture
facilities, technical skill in utilizing sterile techniques and the ability
to distinguish maternal haploids from aneuploids in the seedling stage.
Difficulties are often encountered in obtaining haploids and their
frequency is low even under good conditions. The efficiency of the
doubled haploid method relative to other methods is questionable for
those populations where the frequency of desired genotypes is
expected to be low.
e
Interspecific Hybridization
Interspecific hybridization procedures generally require doubling the
chromosome complement of one or both of the parents or their hybrid
to insure fertile offspring. This is necessary because the two parents
may have unequal chromosome numbers with little chromosome
homology, and without polyploidization no viable gametes are
formed. In some instances, usually when two species are very
distantly related, no viable seeds are formed even with chromosome
doubling. It has been possible, in some of these cases, to accomplish
hybridization by first crossing one of the species to a third species
(bridge cross). Protoplast fusion has also been used to produce hybrids
between some intractable parental combinations, but this technique
has yet to result in the transfer of useful traits to N. tabacum. Once
viable interspecific plants have been grown to flowering, they are
backcrossed to the recurrent parent (usually N. tabacum) to start
eliminating unwanted chromosomes and genes of the donor species.
Because of the lack of chromosome pairing in the initial breakdown
generations, normal crossing over and gene transfers are rare and
random events. Often a whole alien chromosome is substituted while
chromosome additions or segmental substitutions may also occur. In
all cases, unwanted and often undesirable genes are transferred in
tandem via linkage to the gene controlling the desired trait. Where the
donor species is closely related to N. tabacum, some degree of
crossing over between homologous chromosomes will occur and
transfer of recessive genes is possible. Unfortunately, most of the
species with resistance to major diseases are not closely related to N.
tabacum and only genes with dominant or partially dominant
expression can be selected through segregating generations. Burk and
Chaplin (1979) have reviewed procedures for interspecific
hybridization.
f
Recurrent Selection
Recurrent selection procedures offer breeders a way to maximize the
amount of improvement from a given population. Increasing
opportunities for genetic recombination through intermating allow
greater opportunity to exploit all the genetic diversity among the
parental genotypes of a population. Several studies

 Page 44
with flue-cured tobacco demonstrate the potential of recurrent
selection in tobacco improvement, especially in increasing the
frequency of desirable genes in a breeding population (Matzinger &
Wernsman, 1979).
The negative relationship between yield and nicotine has limited the
improvement of cultivars for yield. Many high-yielding lines have not
been considered for commercial release because of low nicotine
levels. In attempting to obviate this problem, Matzinger, et al. (1972)
combined a restricted selection index and recurrent selection so that
only high-yielding plants with acceptable nicotine were intercrossed
to create the next cycle. Under this scheme the rate of yield
improvement was reduced to half that expected from selection for
yield per se.
g
Genetic Engineering
Research in genetic engineering (see following chapter) has provided,
and will likely continue to provide, new techniques for use in plant
improvement. New technologies are changing tobacco breeding
methods, especially those required to utilize diverse sources of
germplasm. Classic breeding techniques such as chromosome
doubling, bridge crossing and extensive backcrossing are no longer
required to achieve interspecific gene transfers. The role and perhaps
even the need for standard procedures such as the pedigree and
backcross methods are changing.
As genes are transferred into tobacco from other Nicotiana species
and perhaps other plants and animals, the characteristics of the
derived plants will be very important in determining their use by
breeders. Cultivars must have many desirable characteristics that
include disease resistance, good handling properties, acceptable yields
of high quality leaf and acceptable concentrations of several chemical
constituents. The amount of manipulation required to get from a
genetically-engineered plant to a cultivar will undoubtedly require
considerable effort. As discussed earlier in this chapter, the phenotype
of a plant is determined by the sum of all its genetic material and the
environment in which it is grown. Genetically-engineered lines will
probably require extensive testing to identify phenotypes with good
expression of the engineered gene and overall acceptable
characteristics.
Evaluation of Genotypes
Dependable techniques for the evaluation of plants and lines are
required in both genetic studies and breeding programs. Specific
aspects of screening and testing techniques will depend upon various
factors such as objectives, genetic materials, available resources and
technical expertise. The following discussion is centered around
techniques that breeders and geneticists have found from research and
through experience to be acceptable in many situations.
a
Disease Screening Techniques
A detailed account about maintenance of disease cultures, media
preparation, inoculation techniques and screening procedures has been
given for most of the diseases of tobacco (Stavely, 1979). Use of
greenhouses for growing and screening plants is advantageous in
respect to time, labor and symptom expression for many diseases
especially when small seedlings can be tested. Screening of seedlings
for black root rot, wildfire and viruses is quite accurate, and breeders
routinely use greenhouse facilities to evaluate genetic material for
reaction to these diseases. A technique using detached leaves has been
developed which makes it feasible to screen greenhouse plants for
rootknot nematodes, tobacco mosaic virus and wildfire (Rufty, et al.,
1986). Since the technique does not introduce disease organisms into
the plants, seed can be obtained from the tested plants. In addition, the
technique can be combined with field testing for reaction to other
diseases or for evaluation of agronomic and chemical characteristics.
When large plants are required or expression of disease resistance is
poor under greenhouse conditions, field screening is usually the most
economical means of evaluating the large numbers of plants needed in
a breeding program. For example, greenhouse screening for diseases
like bacterial wilt and black shank often gives variable results, from
complete kill of resistant plants to no symptoms on susceptible plants.
Moderate to high levels of resistance, which are adequate in most field
situations, are usually difficult to detect in greenhouse tests. Results of
screening for black shank resistance are often variable under field
conditions, but the use of naturally or artificially infected fields gives
useful information, especially when there is a systematic placement of
check rows consisting of susceptible material within the test area.
These check rows give the breeder a reference upon which to judge
the symptoms or lack of symptoms on the test progenies.
Fields with a history of disease are also useful in testing for resistance
to bacterial and Fusarium wilts and nematodes. Artificial inoculations
at transplanting can enhance the chances of getting more uniform

 Page 45
infection, and thus reduce the frequency of selecting susceptible plants
because they have escaped infection. Testing for blue mold reaction is
best done in areas where natural outbreaks occur on a consistent basis
year after year.
b
Field Testing Procedures
The evaluation of genetic materials for agronomic performance and
chemical composition in both genetic studies and breeding programs
requires field testing. A host of factors must be considered when
planning a field test. Consistent, reliable and accurate techniques are
an absolute requirement to reduce nongenetic variation and are the
responsibility of the experimenter or breeder. One should strive to
maintain uniformity in field operations so that experimental entries
grow under comparable conditions. Errors in such procedures as
taking measurements, weights and chemical samples should be kept to
a minimum. Leaf loss during field operations, harvesting, curing and
stripping can cause large experimental errors and false differences
among entries if not adequately controlled. In addition to the use of
good techniques, choice of experimental design, number of
replications and plot size are used to reduce the magnitude of both
random and unavoidable sources of variation. Repetition of tests at
more than one location and in more than one year is done to measure
genotype × environment interactions. Fortunately, genotype ×
environment interactions in tobacco are relatively small compared to
genotypic effects. Thus, tests can be conducted in a limited number of
locations and during one or two years. Repetition of tests in more than
one environment does give the experimenter some insurance against
loss due to some uncontrollable factor such as hail and provides a way
to detect the effects of unusual weather at a specific location during
any given year.
In the United States, tobacco breeders and geneticists have found the
randomized complete block design adequate for most situations. For
genetic studies and initial testing of breeding lines, plots are generally
single rows consisting of 22 to 35 plants. For flue-cured where leaves
are removed from the plants for curing, 20 competitive plants have
been used per plot. In burley areas, the clay soils are more
heterogeneous and stalk cutting tends to cause more leaf damage
during harvest. Thus, harvesting 30 competitive plants per plot has
been practised. Two and sometimes three replications have been used
in genetic studies designed to evaluate agronomic and chemical traits.
Scientists generally feel that more information is obtained by
maximizing the number of genotypes and minimizing the number of
replications and test environments.
c
Advanced Line Testing
The end result of a breeding project is usually a large array of lines
and limited information on their performance for many important
traits. The most economical way to reduce the number of lines and
identify cultivar candidates, if any, is to conduct a series of
performance trials starting with preliminary tests to evaluate growth
properties and ending with tests throughout the production area to
determine on-farm performance, to assay leaf physical and chemical
properties and to determine whether flavor components are
acceptable.
Wernsman and Rufty (1987) have outlined in considerable detail the
testing procedures used for flue-cured tobacco in the United States.
This program has worked satisfactorily and involves all segments of
the tobacco trade including growers, leaf dealers, manufacturers,
private tobacco breeders and scientists from public institutions. The
system provides reasonable assurance that new cultivars will meet an
acceptable standard of performance for the most important attributes
under diverse cultural and environmental conditions. This should be
the primary objective in the evaluation of breeding lines. Key
elements of a testing program should be adequate sampling of
environmental and cultural conditions encountered in commercial
production, strict adherence to a set of performance standards and
consideration of all important traits especially disease resistance,
quality, chemical traits and flavor components. The inclusion of check
cultivars in the testing program provides an acceptable standard for
judging test lines. For example, personnel in the USA flue-cured
testing program consider a breeding line to be acceptable in nicotine
concentration if it falls within the range of -20 to +15% of the mean of
two check cultivars, NC 95 and NC 2326.
Breeding Progress
Genetic improvements have been made in a wide range of
characteristics including pest resistance, physical attributes,
agronomic performance, leaf quality and chemical composition of
cured leaf. The amount of progress for most traits is difficult to assess
because many of the factors that must be considered in developing an
evaluation criterion change with time. Such factors include
improvements in production tech-

 Page 46
nology, environmental and pest conditions faced by tobacco growers
and chemical characteristics of the cured leaf needed by
manufacturers to produce consumer products.
Contributions made in breeding cultivars for disease resistance are
especially noteworthy and quite significant. Cultivars with high levels
of resistance are available in many of the tobacco classes. Also,
resistant cultivars have provided the most economical and perhaps the
only feasible means of controlling certain diseases in some production
areas. For example, the benefits of black root rot resistant cultivars of
dark fire-cured tobacco in Canada have been spectacular, representing
the difference between a good harvest and crop failure. Such
stabilizing effects on production are a very important feature of
resistant cultivars. Reasonable harvests every year are much preferred
to the economic hardships associated with extreme fluctuations in
yield.
Yield improvements have been hampered by the negative relationship
that exists with nicotine concentration. Nevertheless, progress has
been made as evidenced by the comparative yield of modern cultivars
with those in production 30 years ago. Bowman, et al. (1984)
compared the yield of a flue-cured cultivar, Hicks, with newer
cultivars entered in the North Carolina Official Variety Test between
1954 and 1981 and estimated that the average annual yield increase
due to genetic improvement was approximately 16 kg/ha. This
increase was accomplished with only a small decline in nicotine
levels. Quality, as measured in dollar value per hundred kilograms,
also showed significant improvement with an average annual increase
of 26 cents/100 kg.
Limited breeding progress has been made in altering the levels of
smoke constituents and flavor components. Many require laborious
analytical procedures which limits the number of plants a breeder can
assay in segregating populations for desirable chemical traits. Also, it
is often unclear how these constituents affect the sensory properties of
the leaf. However, most of the chemical constituents of the leaf (and
many morphological traits such as leaf shape which indirectly affect
leaf chemistry) contribute to smoke chemistry, flavor and aroma.
Their specific effects are often obscure because most are altered by
pyrolysis. In addition, the ratio between different constituents such as
sugar and nicotine may be more important than each component per
se.
The total particulate matter (TPM) produced by a burning cigarette
(commonly known as tar) can be altered to some extent by selection.
Matzinger, et al. (1989) measured the particulate matter index (PMI),
which gives an estimate of tar, in inbred lines derived from a
population after five cycles of recurrent selection for lower PMI. The
line with the lowest PMI was only 13.7% below the mean of the two
parents. It would seem that reducing tar through breeding techniques
will be an arduous task.
Conclusions
There have been many advances in tobacco breeding and genetics
over the past several years. Breeders have developed cultivars with
high yield, good quality and resistance to several diseases. In addition,
inheritance information has been obtained for numerous
characteristics, and a vast array of germplasm has been collected and
characterized. This solid base of genetic information and germplasm
enhances the prospects for future advancements. Also, new
information and emerging technologies in chemistry and biology,
especially in genetic engineering, have promising applications in
tobacco improvement.
To utilize scientific advances effectively, breeders will need to keep
up-to-date on new information and technologies and make appropriate
changes in their breeding programs. Objectives may need changing as
production practices are altered, different pests pose new threats,
improved germplasm and cultivars are released and consumer
preferences change. Breeding methods will likely need modifying
rather dramatically as genetic engineering and other laboratory
procedures are used to change the plant genetically. The cooperation
of tobacco breeders with scientists from other agricultural and
biological disciplines will be very important to achieve maximum
progress in cultivar development.
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25774.

 Page 49

Chapter 3
Biotechnology: Uses and Applications in Tobacco
Improvement
J.D, Brandle
Agriculture and Agri-Food
Delhi, Ontario, Canada

D. Bai
Imperial Tobacco Ltd
Montreal, Quebec, Canada
Introduction
Since the 1970s the development of new molecular genetic techniques
such as plant transformation, genetic mapping with molecular
markers, gene tagging and positional cloning has significantly
improved the possibility of overcoming the constraints that have
hindered, for many years, the improvement of tobacco (Nicotiana
tabacum) pest resistance, yield and quality. Traditionally, the only
source of new genes for tobacco improvement has been tobacco itself
and, to some extent, the wild relatives. Plant transformation now
allows the introgression of genes from virtually any organism and is
leading to the creation of exotic phenotypes with many applications in
crop production. Furthermore, the mapping and analysis of plant
genomes is providing the foundation for the isolation of genes using
positional cloning. Transposon and TDNA tagging is allowing the
characterization of many genes involved in disease resistance and
plant development. Now, the difficult problems that have remained
despite many years of breeding efforts can be approached with these
new tools in hand. As a result new cultivars that make more efficient
use of production resources and generate more marketable products
will be developed. The purpose of this review is to discuss the
application of these techniques, to define their role in a tobacco
breeding program and to identify existing systems that could be used
in tobacco improvement.
Genome Analysis
The analysis of plant genomes starts with the development of a
traditional genetic linkage map that uses molecular markers. This is
followed by physical mapping (sequencing), which eventually leads to
the complete DNA sequence of the whole genome. Functions are then
assigned to the various coding regions and an understanding of the
organization and operation of the plant geneome is developed. From
any perspective, the tobacco genome is large, in the area of 1600
megabase pairs, of which approximately 75% is repetitive or
noncoding. The repetitive DNA consists of tandem repeats (satellites,
DNA, telomeres) and dispersed repeated sequences (tetrotransposons,
long interspersed repeats, transposable elements) (Dean & Schmidt,
1995). Estimates suggest that about 27000 different structural genes
are expressed in tobacco leaves at any one time, and when one
considers those genes unique to certain tissues or developmental
stages, the total number is probably closer to 60 000 (Goldberg, et al.,
1978; Kamalay & Goldberg, 1980). A recent search of the
nonredundant nucleotide database at National Center for
Biotechnological Information revealed that there were 677 tobacco
sequences, of which 572 were from nuclear genes, clearly
demonstrating that a great deal of the tobacco nuclear genome remains
unexplored. The complete chloroplast genome of tobacco was
sequenced in 1986 (Shinozake, et al. 1986). Arabidopsis, on the other
hand, has a much smaller genome, estimated at 70


 Page 50
Mbp and 25 000 genes (Flavell, 1980; Meyerowitz & Pruitt, 1985)
and of that more than 8000 genes have been sequenced or partially
sequenced (Newman, et al., 1994).
A genetic linkage map is a linear array of molecular or classical
markers that represents the relative position of each marker. A
molecular marker is a DNA fragment that is unique in terms of size
and sequence. Assuming a genome length of 1000 cM (centi-
Morgan), 1600 markers would be necessary to map the large tobacco
genome to one megabasepair resolution. At this time no tobacco
genetic map, either classical or molecular, exists so it would be
necessary to start from the beginning. Furthermore, to be truly useful,
markers should be placed at 400 kb intervals and to have a 90%
chance of having a marker within 400 kb of a given target gene at
least 7500 markers should be sampled. The development of such a
high density map would require a concerted effort and is probably not
necessary. Nonetheless, the contributions made by these genetic maps
have been enormous, they have led to a deeper understanding of
genome organization within a species and allowed the comparison of
genomic organization among crop species. It has been shown that
large stretches of genomic DNA are collinear between related crop
species, such as potato (Solanum tuberosum) and tomato
(Lycopersicon esculentum) (Tanksley, et al., 1992). Since tobacco is
related to both tomato and potato, genes isolated in those species
could be directly transferred to tobacco and may prove to be a great
resource in tobacco improvement. In addition, different types of
molecular markers such as restriction fragment length polymorphism
(RFLP), randomly amplified polymorphic DNA (RAPD), amplified
fragment length polymorphism (AFLP) and simple sequence repeats
(SSR) have been used (Tanksley, et al., 1992; Williams, et al., 1993;
Thomas, et al., 1994; Powell, et al., 1996; Staub, et al., 1996).
Variations of those marker types in combination with genetic
materials such as F2 bulks (Michelmore, et al., 1991) or near isogenic
lines (Martin, et al., 1991) have uncovered linkages to many
agronomically significant genes. In our experience with RAPD, AFLP
and SSR markers systems, tobacco is not highly polymorphic
(unpublished data).
In tomato, the molecular linkage map has been used in many ways to
advance the understanding of that species. Tanksley, using tomato as a
model, has listed three general categories of applications for
molecular linkage maps (Tanksley, 1993). The first is
population/evolutionary studies, such as that of Miller and Tanksley
(1990), of the variation within and among species in the genus
Lycopersicon. They confirmed the phylogenetic relationships among
the tomato relatives and demonstrated the narrow genetic base that
forms the foundation of modern tomato breeding. This may be true,
generally, in tobacco and more likely true for flue-cured tobacco. The
second application is gene tagging, where molecular markers linked to
qualitative or quantitative (QTL) genes are identified and used as a
selection tool (Osborn, et al., 1987; Paterson, et al., 1988). Molecular
markers represent the genotype of a plant and are not subject to the
vagaries of environmental variation that are normally found with
phenotypic selection. Currently many molecular markers have been
found that are tightly linked to important plant genes, including two
RAPD markers found linked to the 'Burley 49' gene for black root rot
(Chalara elegans) resistance (Bai, et al., 1996). Selection for visual
quality in flue-cured tobacco could be greatly assisted by the
availability of genetic markers linked to QTLs influencing those traits.
Computer simulation studies have shown that marker assisted
selection (MAS) can speed up the process of selection for quantitative
traits, by allowing two cycles of selection per year (Edwards & Page,
1994). The same applies for simple monogenic traits where MAS can
eliminate the need for progeny testing and phenotypic screening. The
third application is positional cloning, where detailed molecular
genetic maps are essential. A recent example of this approach is the
isolation of the Arabidopsis RPS2 gene for resistance to Pseudomonas
syringae, a significant pathogen of tomato (Bent, et al., 1994).
Restriction Fragment Length Polymorphism (RFLP)
A number of agricultural species have been mapped using RFLPs
(Tanksley, et al., 1989). As far as the utility of RFLP markers as a
selection tool is concerned, the process requires too much DNA and is
too impractical to be of great use to an applied plant breeder, at least
for simple monogenic traits such as pest resistance and for selection in
early generations where plant material may be limited. However, once
linked markers are found they can be converted to SCARs (sequence
characterized amplified regions) and simpler, polymerase chain
reaction (PCR) based methods can be used for selection (Paran &
Michelmore, 1993).

 Page 51
Randomly Amplified Polymorphic DNA (RAPD)
Randomly amplified polymorphic DNA markers are generated by
mixing a single oligonucleotide of arbitrary sequence (primer) with
genomic DNA, a thermostable DNA polymerase, dNTPs and a buffer.
The mixture is then subjected to temperature cycling conditions and
the process is called PCR. The single primer binds to specific
homologous sites on opposite strands of the genomic DNA and, if the
primers are at an amplifiable distance (a maximum of about 3000
nucleotides), the polymerase proceeds across the region between the
two primers and the result is a discrete DNA fragment. Since the
extension products are complementary to and can bind to the primer,
further cycles in the amplification double the amount of the fragment
produced in the previous cycle and its concentration increases
exponentially. The amplified fragments are separated on agarose gels,
visualized by ethidium bromide staining, and polymorphisms between
the various genomic DNA samples are scored. In a similar vein, RFLP
markers or other sequence characterized fragments can be amplified
using PCR. No radiolabelled probes and only nanogram quantities of
genomic DNA are required, and the work can be completed in one
day. When combined with rapid genomic DNA extraction procedures
such as the seedling leaf procedure, which is a modification of
Edwards, et al. (1991), where 150 extractions can be done in one day,
a large F2 population can be easily scored for a linked marker over the
course of a few days. Costs of the two procedures are similar and
unlike most PCR procedures no prior knowledge of target sequences
is required. New procedures that are even faster have been proposed
(Gu, et al., 1995).
RAPD markers are dominant and it is therefore not possible to
distinguish heterozygous genotypes from homozygous when mapping
F2 populations. This is not a problem, however, when using
recombinant inbred lines as the mapping population since such lines
are fixed for the gene in question during the inbreeding process. A
RAPD fragment can be excised from a gel and used as a probe for
RFLP analysis if codominant markers are necessary and providing the
fragment was amplified from a single copy genomic sequence. An
additional problem with RAPDs is that the genomic origin of the
fragments and sequence homology of bands of similar size are not
known. This latter problem is more significant (Thormann, et al.,
1994). In general, RAPDs can be used in all the applications that were
originally proposed for RFLPs.
Mapping and identifying markers linked to genes significant to leaf
quality and agronomic performance are important for molecular
marker efforts in tobacco. Of immediate interest to plant breeders,
however, are RAPD or SCAR markers linked to disease resistance or
chemical quality genes where screening segregating populations and
progeny testing can be time consuming or expensive. This can be
achieved using materials that exist already in most breeding programs
such as near isogenic lines (NIL) (Martin, et al., 1991) or more simply
by using a technique known as bulk segregant analysis (Michelmore,
et al., 1991) (Fig. 3.1). With NILs, only three genotypes are required:
the donor parent, the NIL and the recurrent parent. Because of this a
large number of primers can be screened in one day and the complete
set can be screened over the course of a few weeks.
Bulk segregant analysis is similarly fast, with an additional advantage
of requiring less time to develop the necessary genetic material, as it
is based on bulked DNA from progeny tested F2 plants segregating for
the gene of interest. Two types of markers can be generated by such a
screening effort, those linked in coupling and those linked in
repulsion. With a single dominant resistance gene, selection using
repulsion phase markers is more efficient as the marker is linked to
the 'susceptible' allele and selection against the marker will result in
lines homozygous for the resistance gene (Haley, et al., 1994). The
first two seedling leaves from a group of F2 are sampled for DNA
extraction, the samples are analyzed using the PCR reaction and then
scored for the absence of the diagnostic marker. A large seedling
population can be screened before transplanting. Selection for other
phenotypic characteristics can then be undertaken in a larger
population of homozygous lines than would normally be present using
standard resistance screening methods. Sixty-six percent of the
material that would be retained using normal phenotypic screening
(e.g. the heterozygotes) is discarded. No progeny testing is necessary
in the F3 generation, saving further time and money. Care must be
taken, however, to ensure that the repulsion phase marker is not
unique to the cross used for the original screening, that is to say the
marker should be present in all potential recipient cultivars and absent
in all donor cultivars. The repulsion phase marker found to be linked
to the Burley 49 gene for black root rot resistance does behave in the
desired way and is therefore well suited to general use as a selection
tool for black root rot resistance in a wide range of crosses (Bai, et al.,
1996). Markers linked in coupling are similarly useful, but it is
usually not

 Page 52
Near Isogenic Lines

Bulked Segregants

RAPD analysis

Fig. 3.1
The development of molecular
markers linked to a dominant
gene for disease resistance
using near isogenic lines (NIL)
or bulk segregants and randomly
amplified polymorphic DNA
(RAPD) marker analysis. Near
isogenic lines are developed by
crossing a donor (P1) and recipient
(P2) cultivar, followed by repeated
backcrossing to the recipient
parent coupled with selection for
disease resistance. The NIL and
the recent parent differ by one
marker (the stipled band), and
it is linked to the disease
resistance gene. For bulked
segregants, the F2 from a cross
 between the donor and recipient
parents for disease resistance and
DNA samples from susceptible
and resistant individuals are
pooled. The BULK1 was selected
for the resistant phenotype and
differs from the BULK2 by only one
marker (the stippled band), that is
linked to the resistance gene
(adapted from Tanksley, et al., 1995).
possible to distinguish homo-and heterozygotes so a further round of
progeny testing that uses marker assisted selection is required. For
backcrossing, markers linked in coupling are ideal and repulsion
phase markers can be used in the final selection of homozygous lines.
Using the method outlined in Bai, et al. (1996), we have screened the
700 decamer primers available from the University of British
Colombia Oligonucleotide Synthesis Laboratory and found that 63%
generated repeatable RAPD fragments. From those primers nearly
2600 fragments ranging in size from 100 to 2000 base pairs were
generated, an average of 5.9 fragments per primer. Readers are
directed to Williams, et al. (1993) for a complete discussion of factors
affecting reproducibility of bands and how variation in the reaction
mixture and conditions can affect the outcome of the assay.
Amplified Fragment Length Polymorphism (AFLP)
Amplified fragment length polymorphism is a newer system that is
based on the selective amplification of restriction digested genomic
DNA, which can result in as many as 50100 bands per reaction
(Thomas, et al., 1994). The ability of this technique to generate large
numbers of high resolution markers in a relatively short period of time
makes it very attractive. This technique is relatively expensive and
more technically demanding than RAPDs.
Simple Sequence Repeats (SSR)
Simple sequence repeats or microsatellites are tandemly repeating
one, two, three, four and five nucleotide units. Primers, designed for
the sequences flanking the repeat, can reveal size polymorphisms that
are codominant, and due to the large primer size the results are more
reproducible than RAPDs (Powell, et al., 1996a; Powell, et al.,
1996b). Prior knowledge of genomic sequence is essential and is the
weakness in this approach. Our work with tobacco sequences
available in the public databases has revealed a number of di-and
trinucleotide repeats within introns and flanking structural genes
(unpublished data). In general, however, levels of polymorphism were
not higher than found with other methods and the discovery of more
SSR primers will require an active screening and sequencing effort.
Transformation
Transformation is a technique that allows the introduction and
expression of foreign genes into plant species. The foreign genes
confer unique traits, not normally found with the species, upon the
recipient plant. Plant breeding has been dramatically strength-

 Page 53
ened by the development of plant transformation and the
accompanying understanding of gene structure and function. The
barriers to interspecific gene transfer that prevented the introgression
of genes from even closely related species have been significantly
weakened, and it is now possible to express genes from animals,
viruses, bacteria and unrelated plant species in several significant
crops including tobacco. The goals of this new technology are not
different from those of classical plant breeding and when the two are
combined they are highly synergistic. Tobacco was the model system
used to develop a large part of this technology, but despite this
pioneering role much of the application of this technology is now
occurring in other crops. Ahl Goy and Duesing (1995) reported that,
in 1994, tobacco accounted for 12.5% of all transgenic field trials
conducted worldwide, down from 25% a few years earlier. While the
authors concluded that there was a significant trend away from
tobacco, it is more probable that the reduction reflects a more mature
research focus, where trials are now limited to those with a more
direct impact on tobacco production. This is in contrast to the 'look
and see' type of work that was done in the model system era of
tobacco transgenics. A time course comparison of the ratio of trials
conducted with commercial tobacco varieties versus those done with
SR1 and similar lab varieties would shed light on the real nature of
this trend. There are a large number of genes that could be used to
solve tobacco improvement problems, such as blue mold and insect
resistance, that have not been solved by conventional efforts for years.
It is now only a question of adding transformation to the germplasm
development component of the breeding program. The success of this
depends heavily on the establishment of a strong collaborative
research effort involving molecular geneticists, breeders, pathologists,
virologists, weed scientists and entomologists.
The current focus in transgenic tobacco field trials is crop protection
and is similar to that of the other major crops such as maize, canola
and potato (Ahl Goy & Duesing, 1995). Trials include insect, virus
and fungal disease resistance, and herbicide resistance. There is
potential for the development of systems that will allow the
modification (either removal or addition) of plant secondary
metabolites and the addition of 'value added traits'. This has been
described in a review article (Gadani, et al., 1995). Currently, traits
introduced by transformation are generally restricted to single genes
and the direct modification of polygenic traits such as yield is not
possible; however, the advent of transformation with high molecular
weight DNA means that the introduction of very large gene clusters is
feasible in the near future (Hamilton, et al., 1996).
a
Agrobacterium Mediated Transformation
In tobacco, Agrobacterium mediated transformation is the gene
transfer method of choice. It is fast, relatively simple and reliable.
Agrobacterium infects wounded plant cells, leading to the production
of tumorous growth at the site of infection. The tumor cells result
from the transfer of a piece of DNA known as the T-DNA from the
bacterium to the plant. Agrobacterium carries a large plasmid (Ti
plasmid) on which the T-DNA and the genes (vir region) involved in
the transfer of the T-DNA are located (Zambryski, et al., 1989).
Normally the T-DNA carries oncogenes that are responsible for tumor
formation and genes coding for the production of compounds called
opines. Tumorous plant cells produce opines in large amounts which
are used as a substrate for growth of the invading Agrobacterium.
Agrobacterium strains are classified according to the type of opine
that is encoded by their T-DNA. On the left and right sides of the T-
DNA are border sequences that are essential for the transfer of the T-
DNA; however, any sequence between the two border sequences can
be transferred to a plant genome. This discovery coupled with the
observation that the vir genes can act in trans was a major step in
plant genetic manipulation.
Two delivery systems have been developed. In the binary vector
system there are two plasmids in the Agrobacteriumium, one large
plasmid carrying the vir genes, but lacking the T-DNA, and a second
smaller plasmid carrying the T-DNA border sequences, a bacterial
origin of replication, a selectable marker gene and a multicloning site.
This smaller plasmid is easily propagated in E. coli and can be
manipulated so that a plant selectable marker gene and the gene of
interest can be placed between the border sequences. Segments of
foreign DNA as large as 150 kb have been transformed to tobacco
using specialized binary vectors (Hamilton, et al., 1996). A second
system, based on co-integrate vectors, has also been developed and is
in common use. Transformation is achieved by infecting wounded
plant tissue with Agrobacteriumium and then transferring the tissue to
selective media that will allow only stably transformed plant cells to
be regenerated into plantlets (Fig. 3.2). The T-DNA tends to be
integrated into the

 Page 54

Fig. 3.2
Generalized diagram of Agrobacteriumium mediated transformation. Following
activation of the bacteria which leads to attachment to the surface of the plant cell,
the vir genes on the helper plasmid (H) are activated and a new T-DNA strand is
synthesized from the one located on the binary vector (BV). The T-DNA is the
region between the right (RB) and left (LB) border sequences and in this figure
contains two promoter (P) sequences which control the transcription of the foreign
genes and two termination (t) signals which act to stop gene transcription. The
first of the two genes is nptll, which confers resistance, on transformed plant
cells, to the antibiotic kanamycin. It is probably the most common of the plant
selectable markers. The second is the gene of interest (Gol) that confers herbicide
resistance, for example, on the transgenic plants. The new T-DNA, in combination
with two vir proteins that aid in nuclear transport, is then transferred through the
plant cell wall (CW), cell membrane (CM) and nuclear membrane (CN) and
integrated into the plant genome (PC).
plant genome at random, with some preference for actively transcribed
regions (Tinland, 1996).
Opportunities
a
Herbicide Resistance
Research in herbicide resistance has uncovered a number of useful
genes that confer resistance to broad spectrum, high unit activity, low
residue herbicides with low mammalian toxicity. Resistance introduced
into tobacco via transformation increases flexibility by allowing post-
plant application. Herbicide resistance would certainly have a positive
impact on tobacco production, where broad leaf weed control is a
serious problem and where the number of potential herbicides is
limited. Resistance is usually only to one herbicide, and in the absence
of herbicide application the resistance gene is not typically considered
to confer any selective advantage on the crop or to weedy relatives
(Crawley, et al., 1993).
There are three transformation-based strategies that have been used to
create herbicide-resistant crops. The first strategy is to simply over-
express the gene that is the herbicide target; the second is to introduce a
mutant gene which codes for an insensitive version of the enzyme.
When genes are over-expressed, enzyme is produced in excess of what
is required and the application of herbicide has no metabolic impact. In
the second case, the mutant enzyme is insensitive to herbicide action,
yet it retains its normal catalytic properties. When herbicide is applied,
the mutant enzyme simply replaces the function of the wild type
enzyme and a normally sensitive crop remains undamaged. The second
strategy is to metabolically inactivate the herbicide by introducing
genes which code for enzymes that degrade the herbicide into inactive
byproducts. The third strategy is to combine features of both strategies
together in one genetic background.
Direct over-expression of alfalfa glutamine synthetase (GS), the target
enzyme for the herbicide phosphinothricin, was used by Eckes, et al.
(1989) to engineer resistance in tobacco. Glutamine synthetase is
essential for the assimilation of ammonia and, in general, nitrogen
metabolism. Phosphinothricin is a very effective broad spectrum, low
residue herbicide affecting both broad leaf and grassy weeds and crops.
Transgenic tobacco plants over-expressing GS were severely damaged
at four times normal field rates, but eventually grew and reproduced
normally. However, free ammonia levels in leaf tissue were nearly
seven times higher than found in unsprayed controls, clearly an
undesirable side effect, which when coupled with narrow safety
margins made the system commercially unacceptable. Attempts were
made to over-express the target enzyme (EPSP synthase) for the
herbicide glyphosate, and while over-expression was achieved
tolerance to glyphosate at the whole plant levels was not adequate. This
topic was the focus of a review (Hinchee, et al., 1993).
An example of a mutant enzyme insensitive to a herbicide, with
perhaps more successful results, is resistance to sulfonylurea
herbicides. This class of herbicides has particular appeal because of
their low application rates and low mammalian toxicity. Sulfonylureas
act by inhibiting the activity of the enzyme acetolacetate synthase,
which catalyzes a common step in the synthesis of the branch chain
amino acids (valine, leucine and isoleucine). A mutant gene (csr1-1)
that coded for ALS and was insensitive to chlorsulfuron was isolated
from Arabidopsis by Haughn and co-

 Page 55
workers (1988) and was found to confer resistance to the sulfonylurea
herbicide, chlorsulfuron, in transgenic tobacco seedlings. Later, this
gene was introduced into a Canadian flue-cured variety, 'Delgold', in
an attempt to take advantage of the positive properties of
sulfonylureas, to allow post-plant herbicide application, and to
broaden the weed control spectrum (Brandle, et al., 1992). A number
of primary transformants were screened and two lines with high levels
of resistance in greenhouse trials (Brandle, et al., 1992) were selected
for extensive evaluation in field trials (Brandle & Miki, 1993).
Adequate levels of resistance were observed in one of the selections,
but agronomic performance of the two transgenic lines was poor and
resistance levels were not high enough to include a significant margin
of safety. Poor agronomic performance was attributed to somaclonal
variation, insertional mutagenesis or retrotransposon-induced
mutations in the transformants. It was concluded that a larger
population of transgenic lines would allow breeders to select around
performance problems. Later field testing with homozygous lines
derived from the two original primary transformants revealed that the
line showing the best resistance in greenhouse trials suffered from
transgene inactivation and that the condition was induced by
transplanting (Brandle, et al., 1995). A line hemizygous for the same
transgene was not affected and showed uniform, but slightly
inadequate, resistance in field trials.
To improve the system a new mutant gene derived from Brassica
napus was introduced, which had resistance to a larger number of the
sulfonylurea family members, and it was hoped that the problems with
transgene inactivation and agronomic performance could be solved
(Brandle, et al., 1994). The gene was again introduced into Delgold
and the three most resistant lines were selected in greenhouse trials
and then evaluated in plant bed trials and normal yield trials. In field
trials the performance of the untreated lines was equal to
untransformed controls, but the transplanted homozygous lines in the
yield trial again suffered from transgene inactivation (unpublished
data). It is possible that the selection for high levels of resistance in
greenhouse trials leads to expression levels in excess of what can be
tolerated by the plant (Lindbo, et al., 1993). The transgene and
homologous plant transcripts were then targeted for degradation as
part of a self defense mechanism, the net result of which is the loss of
herbicide resistance. If this were indeed the case, it will be difficult to
engineer herbicide resistance based on herbicide insensitive mutant
genes or direct over-expression, because selection for high-expression
levels that are necessary to confer resistance will often lead to
unstable expression patterns.
Bromoxynil resistance is a good example of the successful application
of the metabolic degradation approach. Bromoxynil herbicide is a
photosynthetic inhibitor in plants that acts by binding a part of the
quinone binding complex of photosystem II. A soil bacterium was
discovered that has the ability, via the action of a nitrilase enzyme, to
degrade bromoxynil into its herbicidally inactive primary metabolite
(Stalker, et al., 1988). The bacterial plasmid carrying the gene (bxn)
responsible for this nitrilase activity was isolated, subsequently
subcloned into a plant expression vector and then transformed to
tobacco. The gene was under the control of a light-regulated tissue-
specific promoter (RuBisCO) and resistance at eight times normal
field rates was obtained. Subsequent commercial development has
occurred in air-cured tobacco in France, and agronomic performance
of transgenic lines was equal to untransformed controls and herbicide
resistance was at commercially acceptable levels (Delon, et al., 1994).
The transgenic cultivar has received approval for release without
further need for confined testing (Freyssinet & Derose, 1994).
A gene (bar) coding for an enzyme (phosphinothricin acetyl
transferase, PAT) that detoxifies the herbicide phosphinothricin has
been isolated from the bacterium Streptomyces hygroscopicus.
Transformation experiments and subsequent field experiments with
the bar gene under the control of the 35S promoter demonstrated that
tobacco was tolerant to up to 10 times normal field rates (DeBlock, et
al., 1987). There was no agronomic evaluation of the transgenic
tobaccos included in this experiment and subsequent commercial
application of this resistance has been focused on canola (Brassica
napus). Transgenic canola cultivars with resistance to
phosphinothricin have been approved for uncontrolled release and
commercial production planned in Canada (Anon, 1995a).
The third and final approach, combining detoxification and an
insensitive target enzyme, was used to develop the 'Roundup-Ready'
system of glyphosate tolerance (Anon, 1995b). Initial efforts to
engineer resistance to glyphosate were discussed briefly earlier. Later
efforts were then directed towards the development of glyphosate-
insensitive EPSP synthase. A gene (aroA) coding for a highly resistant
version of EPSPS was isolated from E. coli and, when transformed in
tobacco under the control of the 35S promoter and fused to a
chloroplast transit sequence, acceptable levels of resistance were
obtained (della-Cioppa, et al., 1987; Padgette, et al., 1989).
Subsequent commer-

 Page 56
cialization has occurred in canola; however, the final commercial
glyphosate-resistant cultivars also contain a gene that detoxifies
glyphosate (Anon, 1995b). This second gene is targeted to the
chloroplast and is under control of the 35S promoter.
b
Insect Resistance
There are two systems that could be used for lepidopteran insect
control: the protease inhibitor (PI) system (Hilder, et al., 1987;
Johnson, et al., 1989) and the Bacillus thuringiensis (Bt) system
(Delannay, et al., 1989). While the Bt system gives more effective
control, the PI system may result in better long term insect control
because of lower selection pressure for the development of resistance
in insect populations (Hoffmann, et al., 1992). Protease inhibitors are
part of the natural defense system of plants and are known to act in an
antinutritional way with the insects' digestive enzymes. The primary
effect is to cause a shortage of amino acids, which results from the
insects' inability to digest proteins. Insect bioassays with transgenic
tobacco using the tobacco budworm (Heliothis virescens) and the
cowpea (Vigna unguiculata) trypsin inhibitor gene under the control
of the 35S promoter, as well as the potato PI-II gene and the tobacco
hornworm (Manduca sexta), have demonstrated that resistance to
insect attack is significantly enhanced. Field tests of the CpTI
(Hoffmann, et al., 1992) and PI-II systems (unpublished results) have
demonstrated that the degree of protection is insufficient for
commercial use. This does not mean that the system cannot be
improved as it may be possible to combine different inhibitors to use
improved expression systems or to re-engineer the PI proteins to
increase their effectiveness.
Bacillus thuringiensis is a soil bacterium that accumulates high levels
of insecticidal proteins (d-endotoxins) during sporulation.
Preparations of the fermented bacteria have been used for a number of
years to control lepidopteran insects and are considered to be
'environmentally friendly'. The toxins are highly insect specific and do
not persist after application. There are four main classes of
insecticidal genes in B. thuringiensis; however the cryI genes are the
best understood and have formed the basis for the development of
transgenic crops. When the encoded protein is ingested by insect
larvae, the protein is solubilized and processed into a toxic fragment,
which interacts with specific receptors in the midgut. The result is
paralysis of the mouth parts and the midgut which leads to inhibited
feeding activity and eventual death. This has been summarized in a
review article (Barton & Miller, 1993).
Early attempts to express full length d-endotoxin genes were
unsuccessful, no transgenic plants were recovered and it was
concluded that the full length protein was probably cytotoxic (Barton,
et al., 1987). It was later discovered that truncated forms of the genes
still retain insecticidal activity and that it was possible to express them
in transgenic plants. Expression of these genes in transgenic tobacco
resulted in resistance to tobacco hornworm (Barton, et al., 1987;
Warren, et al., 1992). Expression levels were still too low, and it was
concluded that the problem was related to codon usage. By taking
advantage of degeneracy in the genetic code, synthetic DNA
sequences that more resemble plant sequences were used to replace
the original bacterial sequences, while still retaining the original
peptide sequence. These new synthetic genes led to a 1000-fold
increase in toxin concentration (0.1% of soluble protein) (Perlak, et
al., 1990). Efforts, where the d-endotoxin gene was introduced into
the chloroplast genome, have demonstrated that levels as high as 35%
of soluble protein can be achieved (McBride, et al., 1995).
The major questions that remain are the development of d-endotoxin-
resistant insect populations and safety as it relates to human
consumption. A number of management strategies aimed at curbing
insect resistance in transgenic crops have been proposed (McGaughey
& Whalon, 1992). The four main strategies are:
(1) deployment of multiple Bt genes, each of which produce d-
endotoxin that acts on a different receptor;
(2) the use of inducible or tissue specific promoters;
(3) the high dose approach where toxin levels are extremely high and
escapes are limited, and
(4) field based methods such as patchwork fields, gene rotation and
refuges.
None offer clear advantages in every situation. Of the first three
genetic strategies all have been achieved, the expression of multiple d-
endotoxin genes (Vandersalm, et al., 1994), tissue specific expression
(Koziel, et al., 1993), high dose expression (McBride, et al., 1995). A
genetic 'on-off' switch has also been developed that can be used with
the Bt system (Williams, et al., 1992). Furthermore, computer
simulation studies have demonstrated that patchwork plantings, in
combination with timely planting of nontransgenic refuge fields, can
help delay the development of insect resistance (Alstad & Andow,
1995). Although a number of transgenic crops carrying Bt genes are
set for commercial release (Freyssinet & Derose, 1994), the genes

 Page 57
are targeted to nonconsumed tissues or are in nonfood crops,
therefore, regulatory approval for human consumption has yet to be
addressed.
Enhanced resistance to the peach aphid (Myzus persicae) has been
reported in transgenic tobacco expressing a lectin gene from
snowdrop (Galanthus nivalis) (Hilder, et al., 1995). Lectins are
carbohydrate binding proteins; leaf disc and whole plant bioassays
have demonstrated that the system may have potential for aphid
control. Impaired aphid nymph development has been reported on
transgenic N. plumbaginifolia expressing a bacterial ipt (isopenetenyl
transferase) gene (Smigocki, et al., 1993). The authors attributed the
resistance to changes in secondary metabolism.
c
Nematode Resistance
Three strategies have been employed to effect control of plant
parasitic nematodes using genetic engineering (Williamson & Hussey,
1996). One of the systems is based on feeding site specific gene
expression coupled with induced cell death, a type of engineered
hypersensitive response. The strategy seems promising, although no
published examples have yet appeared. The second approach is to
express genes that produce proteins that are nematicidal or prevent
infection. Urwin and co-workers (1995) expressed an engineered
cysteine protease inhibitor (OcIDD86) from rice (Oryza sativa L.) in
transgenic tomato and found that it had a significantly negative effect
on the growth and development of the potato cyst nematode
(Globodera pallida). The engineered protein had a single amino acid
deleted which increased its inhibitory activity substantially. A third
method has been the expression of antibodies in tobacco that are
directed against styler secretions essential in the nematode
(Meloidogyne incognita) infection process (Rosso, et al., 1996). Again
the technique appears promising, but it has not been evaluated at the
whole plant level. High resolution mapping of nematode resistance
genes is underway as well, but the cloning of those genes has not been
reported.
d
Fungal and Bacterial Disease Resistance
In large measure, the initial efforts in this area have been devoted to
the development of general or horizontal resistance in transgenic
crops. Now that a number of actual resistance genes have been cloned,
efforts are shifting towards the practical use of those genes and the
isolation of their homologues by less resource intensive methods
(Staub, et al., 1996). The progress in this area has been a direct result
of the molecular characterization of systemic acquired resistance
(SAR) (Brears & Ryals, 1994). Systemic acquired resistance is like
immunization in mammals, in that a general disease-resistant state is
induced following inoculation with pathogens such as viruses. This
resistant state is thought to be the consequence of the expression of
chitinases, glucanases and PR1-like proteins. The first two enzymes
inhibit fungal growth by hydrolyzing fungal cell walls, but the
function of the PR1 proteins is not known (Mauch, et al., 1988).
Overexpression of some of these genes in transgenic plants has
demonstrated that the strategy can provide some protection against a
number of important tobacco pathogens following artificial
inoculation. The efficacy of this approach has yet to be demonstrated
in field trials.
Expression of a bean chitinase gene in transgenic tobacco seedlings
was shown to be effective against seedling damping off (Rhizoctonia
solani) (Broglie, et al., 1991). Combinations of glucanase and
chitinase genes in the same transgenic tobacco line have been shown
to enhance protection against both damping off (Jach, et al., 1995) and
frogeye (Cerospora nicotinae) (Zhou, et al., 1994). Transgenic
tobacco constitutively expressing the PR1a gene from tobacco showed
increased tolerance to both blue mold (Peronospora tabacina) and
black shank (Phytophthora parasitica var nicotinae) (Alexander, et
al., 1993). Constitutive expression of radish (Rhapunus sativus)
antifungal protein 1 was shown to confer some resistance to the foliar
pathogen Alternaria longipes (Terras, et al., 1995). Another
interesting result is the expression of ribosome inactivating proteins
(RIP) in transgenic plants. The RIPs are potent inhibitors of protein
synthesis, yet do not inhibit plant growth when over-expressed.
Expression of RIP alone or in combination with a chitinase gene
effectively reduces the symptoms of damping off (Jach, et al., 1995).
Various other compounds found in plants, such as permatins, lectins
and thionins, have been shown in in vitro assays to have antifungal
activity, but the leap to transgenic plants has not yet been made
(Brears & Ryals, 1994).
On the bacterial side only resistance to wildfire (Pseudomonas
syringae pv tabaci) has been reported. In the first case a gene (ttr) was
isolated from the wildfire organism itself, that acts to confer tolerance
to tabtoxin on P. syringae (Anzai, et al., 1989). It is a so-called self-
resistance gene, that encodes tabtoxin acetyltransferase, an enzyme
involved in the synthesis of tabtoxin. Expression of this gene in
transgenic tobacco resulted in complete resistance to wildfire.
Following the

 Page 58
isolation and cloning of the tomato Pto gene for resistance to bacterial
speck caused by P. syringae pv tomato, the gene was transformed into
tobacco and shown to confer resistance to wildfire (Thilmony, et al.,
1995).
Several plant disease resistance genes have been cloned and shown to
function in heterologous species. Given the fact that many of these
resistance genes share homology, it may then be possible to isolate
more, related resistance genes. Using the original genes to design
degenerate oligonucleotide primers a number of resistance gene
homologs were identified in soybean (Glycine max) and found to map
in a cluster (Kanazin, et al., 1996). If these genes prove to be
resistance genes then the whole cluster could be transformed in
tobacco using new high molecular weight DNA transformation
methods (Hamilton, et al., 1996). As a deeper understanding of the
molecular basis of disease resistance develops, further transformation-
based resistance strategies will be developed and may also provide
good durable resistance to disease problems, such as blue mold, that
have long plagued tobacco crops.
e
Virus Resistance
Transgenic tobaccos carrying virus-derived genes have been shown to
be resistant to related viral diseases and some may provide resistance
to heterologous viruses. These genes include the viral capsid protein
gene, viral replicase subunits, noncoding sequences, complete
genomes of defective interfering viruses and complete genomes of
mild strains of the virus. Fitchen and Beachy (1993) have published a
review article. The mechanism for protection in each of the systems is
not well understood, and the breadth of protection varies from highly
strain specific to broad protection against related viruses. Levels of
virus resistance vary from low to high. For example, coat protein
mediated resistance to PVYN in transgenic tobacco has been very
effective against that strain of the virus and to a significant, but
somewhat lesser extent, against the common strain of PVYO. There
was, however, only limited resistance to other heterologous PVY
strains such as PVYMN and PVYNN (MacDonald, et al., 1997). Of
further interest was the fact that all of the resistant transgenics had
multiple copies of the transgene; however, our segregation analysis
revealed that only one copy was functional in the expression of virus
resistance.
Resistance to tobacco mosaic virus (TMV) in transgenic tobacco
expressing the TMV coat protein (CP) gene was the first reported by
Powell-Abel, et al., (1986). Coat protein mediated resistance has been
demonstrated for a number of significant tobacco viruses including
potato virus Y and potato virus X (Kaniewski, et al., 1990), tobacco
rattle virus (van Dunn & Bol, 1988), tobacco etch virus (Lindbo &
Dougherty, 1992a), tobacco streak virus (van Dunn, et al., 1988), and
tomato spotted wilt virus (MacKenzie & Ellis, 1992). Protection from
both TMV and PVX has been reported in transgenie tobacco
expressing sequences encoding replicase proteins (Golemboski, et al.,
1990; Braun & Hemenway, 1992). RNA mediated resistance was
demonstrated for TEV (Lindbo & Dougherty, 1992b), PVYN (van der
Vlugt, et al., 1992) and TSWV (De Haan, et al., 1992); in all of these
cases, a translationally defective CP gene was used. Both the replicase
protein and RNA mediated resistance is described as 'homology
dependent' and the mechanism is based on the degradation of vital
transcripts by the transgenic plant (Mueller, et al., 1995). In general,
CP mediated resistance is the most popular and can provide high
levels of broad resistance and provides the most promise for the
commercial development of resistant transgenie tobacco.
f
Stress Tolerance
Salt, oxidative and cold stress are major impediments to crop
production. Resistance to ozone, in transgenic tobacco overexpressing
superoxide dismutase (MnSOD), has been reported to improve
tolerance in the ozone sensitive tobacco variety PBD6 by 3-to 4-fold
(Van Camp, et al., 1994). Expression of an E. coli gene (mtlD) coding
for mannitol-1-phosphate dehydrogenase results in the production of
the osmolyte mannitol, which confers salt (NaCl) tolerance on
transgenie tobacco (Tarczynski, et al., 1993). It has been shown that
expression of an Arabidopsis chloroplast w-3 fatty acid desaturase
(FAD7) confers chilling tolerance on transgenie tobacco seedlings at
levels equal to that seen in cold acclimated control seedlings
(Kodama, et al., 1995). Numerous other examples have been cited by
Bohnert and Jensen (1996). While these systems are still at the
prototype stage they provide a starting point from which stress
protection systems can be developed for use in commercial
production.
g
Heavy Metal Tolerance
Tobacco is particularly efficient at absorbing cadmium and early
experiments showed that mammalian metal-

 Page 59
lothionein (MT), a metal scavenging protein, could be expressed in
plants and would confer cellular tolerance to high levels of available
cadmium (Cd) (Lefebvre, et al., 1987). Initial field experiments
conducted in Canada (Brandle, et al., 1993) detected no difference in
Cd levels between transgenic flue-cured tobacco carrying a Chinese
hamster MT gene and untransformed controls. Yeargan, et al. (1993),
however, were able to demonstrate a small, but significant, difference
between transgenic burley, carrying a mouse MT gene, and
untransformed controls in field trials. Elmayan and Tepfer (1994)
expressed human MT in tobacco using the duplicated 35S promoter
and were able to demonstrate substantially reduced Cd levels in
transgenic seedlings. Expression levels in this system were 20-fold
higher than those found by Brandle, et al., (1993) and probably
account for the improvements in the system. The material has not
been field tested to determine if the reduction in Cd levels will be
realized in a normal production environment.
h
Handling Transgenic Plant Material
The choice of which cultivar or breeding line to choose as the
recipient of the transgene is relatively straightforward. Since, in some
respects, the addition of a transgene is similar to backcrossing to
introduce a single gene, some of the same limitations apply. However,
if the breeder wished to complement an existing disease resistance for
example, to provide a more durable resistance, the choice will be
limited to those cultivars carrying the major resistance factors.
Typically, however, one or two elite cultivars or breeding lines are
used. Very few tobacco cultivars are recalcitrant with respect to tissue
culture or transformation by Agrobacteriumium, but some minor
differences do exist (Daub, et al., 1994). The choice can generally be
based solely on agronomic performance and the expectation that the
cultivar already has or will have good grower acceptance and a long
term position in the marketplace. In the Canadian context, a large
portion of the transformation research has been limited to 'Delgold', a
cultivar that has represented more than 50% of the tobacco produced
and has been popular with growers for over 10 years. We have
duplicated much of our research with 'AC Cheng', a newly released
cultivar. To date, none of the transformed varieties in Canada have
been released to growers.
The transformation and regeneration system we use is a modification
of the original system proposed by Horsch, et al. (1985), except that
the explant tissue is taken directly from greenhouse-grown plants and
surface sterilized before inoculation with Agrobacteriumium and the
underlayer of cell culture is no longer used. These modifications save
time and trouble, because aseptic plants and maintenance of cell
cultures are no longer required. One important point is to reduce time
in culture, so that culture induced mutations are minimized in the
derived transgenic lines. Following the regeneration of transformed
plantlets, they should be transferred to rooting media containing the
selection agent corresponding to the selectable marker, which will
ensure that no escapes have been selected.
The number of primary transformants required is hard to estimate as
no data are available, but in our experience it is somewhere between
20 and 50. The frequency with which unstable characters, such as
nornicotine conversion, appear is much higher in material that has
passed through the transformation process. Yield can be negatively
affected, but in both cases insertional mutagenesis caused by the T-
DNA is not the source of variation and the tissue culture process is
implicated (Brandle, unpublished data). The tetraploid tobacco
genome, because of duplication, is better buffered against mutations
resulting from T-DNA insertions. Adequate population sizes will
ensure that both stable gene expression and cultivar performance can
be recovered. This number of transformants is not difficult to achieve
in tobacco. There is a need to analyze fully primary transformants for
gene expression; this can be as simple as spraying herbicide or it may
require a biochemical assay. The relationship between transgene
activity in primary transformants and derived lines has not been
characterized, but the assumption is that heritability will be high. It
should be noted, however, that the relationship between the expression
of the selectable marker gene and expression of the gene of interest is
not always good. So the general routine is to generate the primary
transformants, screen for gene expression following establishment of
plants in the greenhouse and then select the best 20% for creation of
pure lines. Tissue samples (10 g) should be collected from the selected
lines and stored at -70°C for later Southern blot analysis. Pure lines
are developed by collecting self seed from primary transformants,
sterilizing that seed and plating on agar solidified MS media,
containing an antibiotic at a level appropriate to the selectable marker
(for kanamycicn use 150200 mg L-1). Three weeks after germination
the segregation ratios are determined and those having a ratio of 3
resistant:l susceptible can be assumed to have a single insert, a result
that can be confirmed by later Southern blot analysis (Scott, 1988).
Ten antibiotic resistant seedlings are then selected, grown to

 Page 60
flowering and selfed. This seed is then rescreened for antibiotic
resistance and nonsegregating selections tested in field trials,
following analysis of transgene expression. It can be assumed that
there will be no difference between homozygous sister lines derived
from the same primary transformant. The whole process takes about
12 months, so if field trials are planned the first steps must be taken a
year before plant-bed seeding time.
i
Design and Implementation of Field Trials
Experimental designs employed in transgenic field trials are
essentially the same as those normally used in a breeding program and
depend upon the objectives of the experiment, except there is a firm
requirement to compare back to the original parent to ensure that the
phenotype has been fully recovered. Typically, the transgene has no
impact on the agronomic phenotype of the plant and the variation
present among the lines can be ascribed to somaclonal variation.
Transgenic field trials must be designed to fit not only scientific
criteria, but the criteria that must be met for the small scale release of
genetically modified organisms. Generally, the trial plans and the
transgene used are scrutinized by a governmental regulatory agency
and the requirements for trials vary from nation to nation. The
breeder, together with collaborators, may be required to outline in
detail the transgene and its source organism, the nature of the
organism modified, the method used for modification, the nature and
stability of the modification and any potential ecological risk that the
trial may cause. The general principles governing field testing in
many countries and some interesting case studies have been outlined
by van der Meet (1993). The results of early field tests of transgenic
crops have been reviewed by Brandle (1993) and current trends in
field testing presented by Ahl Goy and Duesing (1995).
j
Transgene Inactivation
Instability or silencing in transgene expression is widespread and can
lead to loss of the introduced trait. A number of different mechanisms
have been proposed to explain the phenomenon (Finnegan &
McElroy, 1994; Jorgenson, 1995). At the applied level, the tendency is
to create sufficient transgenic plants and to select around the problem.
Nonetheless, such things as environmentally induced transgene
inactivation can have substantial implications for the practical
application of this technology (Brandle, et al., 1995). This has been a
significant problem in sulfonylurea resistant tobacco lines where a
tandem insert in one of the lines, when homozygous, was found to
stop expressing resistance following transplanting (Brandle, et al.,
1995). Greenhouse and plant-bed-grown plants showed stable
resistance. Several methods have been proposed to avoid this problem
(Finnegan & McElroy, 1994); the simplest is to ensure that only
transgenic lines carrying a single insert are used in field trials and in
subsequent steps to commercialization. The selection procedure
outlined above followed by evaluation by Southern blot analysis
makes this goal achievable. Selection for stable transgene expression
in the production environment will eliminate unstable lines; all that is
required is that sufficient primary transformants are generated to allow
effective selection. During the planning stages leading up to vector
construction, another two points should be considered. First, sequence
homology between independent transgenes, such as the promoter
regions of the selectable marker and the gene of interest, should be
avoided. Second, problems associated with the 'sequence context' of
the transgene may be alleviated by using such things as matrix
attachment regions.
Discovering Plant Genes
A number of established techniques such as library screens based on
cross-hybridization of heterologous probes, library screens with
probes derived from treatment specific RNA populations, screening
expression libraries with antibodies and library screens using
degenerate oligonucleotide probes derived via protein sequencing can
be used to clone genes whose gene products are known. This
discussion will be limited to those situations where the gene products
are not known. These latter methods represent some of the more
current approaches to gene cloning that will provide the source
material for transformation based crop improvement over the next 10
years.
a
Positional Cloning
Positional or map based cloning was first used in plants by Arondel, et
al. (1992) and it is a powerful, but technically demanding, method of
isolating plant genes whose products are not known. The process is
based on the availability of 'libraries' that contain a complete sample
of a species genomic DNA, cut into small manageable pieces that can
be isolated and multiplied.

 Page 61
Bent, et al. (1994) provide a good example of the process.
While it would not be impossible to undertake mapbased cloning in
tobacco, the large genome size requires that a substantial number of
yeast artificial chromosome (YAC) clones be screened. This problem
has, in some respects, been alleviated with the development of
bacterial artificial chromosome (BAC) libraries that are easier to
create and handle than YACs (Woo, et al., 1994). The absence of a
detailed linkage map for tobacco is an additional impediment and
requires regions of interest to be mapped before any positional cloning
work can be started. Martin, et al. (1993) has proposed an alternative
strategy, chromosome landing, and applied it successfully to the
isolation of the Pto gene for Pseudomonas syringae resistance in
tomato. The emphasis in chromosome landing is on isolating DNA
markers that are at a distance from the target gene and less than the
average insert size of the genomic library that will be used for clone
isolation. The target region is saturated with markers using high
volume techniques such as RAPDs and NILs or bulk segregant
analysis. This strategy eliminates the need for chromosome walking
and its associated problems and will likely become the approach of
choice for isolating plant genes (Tanksley, et al., 1995).
b
T-DNA and Transposon Tagging
T-DNA tagging is a powerful technique that is based on the transfer of
the T-DNA of Agrobacteriumium into transcriptionally active regions
of the plant genome. There are two approaches to T-DNA tagging.
The first, passive tagging, is based on the creation of knockout
mutations, which result when the T-DNA disrupts a gene leading to a
recessive mutation. The second, activation tagging, uses
transcriptional enhancers in a specially designed tagging vector. The
plant genes that flank the inserted T-DNA are activated by the
enhancer elements and the result is a dominant or over-expression
mutation. In both cases, the result is a selectable phenotype and the
tagged gene is isolated using probes derived from the tagging vector
(Walden, et al., 1991; Hayashi, et al., 1992; Koncz, et al., 1992).
Activation tagging has been used to isolate tobacco genes involved in
auxin signal transduction and the regulation of polyamine biosynthesis
(Hayashi, et al., 1992; Fritzlar, et al., 1995).
Transposon tagging is a passive system and is based on the creation of
knockout mutations through maize transposon activity (Balcells, et
al., 1991). The general principle is the same as passive T-DNA
tagging, except that transposons, mobile genetic elements that can
excise from one place in the genome and insert in another, are used.
An example of this technique is the isolation of the N. glutinosa gene
for TMV resistance from tobacco (Barton, et al., 1987). This gene can
now be used, via plant transformation, in tobacco improvement
without the flanking N. glutinosa sequences that may have caused the
poor agronomic performance of conventionally derived varieties
carrying this gene. While positional cloning and gene tagging may not
find their way directly into an applied breeding program, the results of
many of these gene isolation experiments (e.g. TMV resistance) can
be used if the capability for plant transformation exists in the breeding
program.
Conclusion
Plant molecular genetics is rapidly evolving. The products of this
research and the new tools developed are an enormous resource that
can be easily tapped by plant breeders. It is now the time to integrate
these new techniques into classical breeding programs and use them
as routine alternatives or adjuncts to conventional breeding
approaches. The combination of new and old will remove yield and
quality barriers and lead to unforeseen improvements in production
efficiency.
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Chapter 4
Agronomy and Physiology
4A
Tobacco Seed
T.W. Hutchens
F.W. Rickard Seeds, Inc.
Lexington, Kentucky, USA
Introduction
Tobacco has perhaps the smallest seed of all the major horticultural
and agronomic crops. Indeed, tobacco seed is so tiny that there are
approximately 10000 seed per gram or 10 million per kilogram. This
small seed size and the fact that the tobacco plant is a prolific
producer of seed have combined to discourage much research on
tobacco seed development and seed quality. Now, however, rapid
changes and advancements in transplant production technologies have
placed a much greater importance on understanding seed development
and those factors that may affect seed quality as measured by
germination rates and seedling vigor. This chapter section will review
the current state of knowledge of tobacco seed production and link
that information to seed.quality characteristics.
Flowering and Seed Development
Unlike most tobacco that is grown for leaf production, tobacco plants
grown for seed are not topped and flower development and seed
maturation are closely monitored. The first stage of flowering is the
development of the flower primordia. In most commercial tobacco
varieties, this initiation is not affected by day length, but rather it is
developmentally regulated. The only commercial varieties of the
major tobacco types that are daylength sensitive are those flue-cured
varieties known as non flowering. These genotypes will produce
flowers, but flower initiation occurs only after the night period
lengthens later in thegrowing season. Virtually all other tobacco
genotypes flower between 55 and 80 days after transplanting. The
actual time between flowering and transplanting varies according to
the variety and environmental conditions. Periods of dry weather or
excessively cold or hot temperatures often delay plant development
and, consequently, the time from transplanting to flowering also is
delayed.
The primary meristem of tobacco terminates in a large inflorescence
or flower structure capable of producing hundreds of individual
flowers. A single terminal flower is usually produced at the center of
the inflorescence, and numerous additional flowers are produced on
branches for as long as several weeks to 3 months. As the
inflorescence matures, plants will initiate growth in axillary buds or
suckers located near the top of the plant. These suckers will flower
and can produce as much seed as the central inflorescence.
Tobacco flowers are classified as perfect and consist of a corolla 6 to
10 cm in length with the pistil and stamens enclosed within the
corolla. Once reaching a development stage where the corolla is as
long as the calyx, flower development proceeds quite rapidly. In as
short as 2 to 5 days, a fully developed flower has emerged and is
ready for pollination. Tobacco is classified as a self-pollinated species
and, often, anthers dehisce or release the pollen while the corolla is
still closed. Pollen is not windborne, and the frequency of cross-
pollination is often less than 5%. Litton and Stokes (1964) suggested
that the amount of cross-pollination is affected by environmental
conditions, genetic differences among varieties and the activity of
pollen vectors such as bees and hummingbirds.


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Pollen germinates in the moist environment found on the surface of
the stigma. The germ tube penetrates the stigmatic surface and grows
several centimeters to the unfertilized ovules. Depending upon
environmental conditions and the physiological status of the stigma
and style, fertilization occurs within 4 days after pollination. Stigmas
may be receptive to pollen over a period of several days, but without
pollination, flowers are aborted and drop from the plant. Under
favorable growing conditions, tobacco seed is physiologically mature
21 to 24 days after fertilization. Although Gwynn (1973)
demonstrated that immature seed removed from the developing seed
capsule can germinate in as little as 10 days after pollination,
germination rates and vigor of seed harvested early are greatly
reduced. Physiologically mature seed does require a drying period
before it can be harvested and stored.
A single seed capsule can produce as many as 3000 seed, and with the
possibility of several hundred flowers per plant, more than a million
seed can be harvested from a single plant. However, the quantity of
seed produced by an individual plant is affected by a number of
factors including the variety used and general growing conditions. For
instance, nitrogen fertilization of the mother plant can affect seed
germination properties (Thomas & Raper, 1979). However, a general
rule can be applied that states that varieties with vigorous leaf
development are good seed producers. Since seed develops over a
period from 6 weeks to 3 months, the quantity and quality of seed
produced are affected by the same environmental stresses extreme
temperature, water deficits or excesses, poor mineral nutrition, insects
and disease that affect tobacco grown for leaf.
Seed Quality
The percent germination obtained in a particular lot of tobacco seed
has been the traditional measure of seed quality. This has proven
adequate for traditional methods of transplant production where as
many as five times the number of seed are sown for the number of
transplants needed. With the development of more sophisticated
transplant systems (see next section of this chapter), such as container-
grown transplants, float systems and precision seeded beds, our
traditional thinking about seed quality has been challenged.
Undoubtedly, these new, higher technology transplant systems will
require the use of seed with high germination rates and increased
vigor.
Requirements for Seed Germination
a
Light
Using conventional transplant production techniques, tobacco seed is
sown directly on the soil surface where it is exposed to light. More
recently, new transplant production methods require a larger seed that
can be handled individually. This is achieved by coating the seed with
an opaque material, thereby limiting exposure to light until the coating
material is split or disintegrates following water uptake. Kasperbauer
(1968), and others (Spaulding & Steffens, 1969; Mohapatra &
Johnson, 1978), have described light-requiring and light-indifferent
characteristics for tobacco seed germination. Kasperbauer (1968)
suggested that sensitivity to light may be inherited and temperature-
dependent. However, combining these reports with empirical data of
our own suggests that the light-requiring characteristic can be found in
all types of seed. Indeed, one lot of a particular variety can contain
seed that requires light for germination while other seeds in the same
lot are light-indifferent. Light-requiring seed can be described as
photo-dormant, and this photo-dormancy has been observed to be
transient and present to a large extent in freshly-harvested seed. Seed
stored under proper conditions for 3 months after harvest usually loses
some photo-dormancy, but one method to ensure that the seed is not
photo-dormant at sowing time is to use seed that has been primed
(described in more detail later in this section) prior to coating.
b
Moisture
The uptake and absorption of water is essential for the onset of seed
germination. The physiological and biochemical changes associated
with seed germination will not be discussed in this section, but it
should be noted that because tobacco seed is so small and cannot store
very much moisture, it is very vulnerable to changes in the supply of
moisture. After sufficient moisture has been made available to initiate
germination, the most critical time is the period from when the radicle
emerges to when it penetrates the soil sufficiently to access the
moisture supply. At this time, a change in moisture availability from a
lack of rainfall or no irrigation, winds crusting the soil or excessive
evaporation caused by high temperatures can result in poor
germination. A germinating tobacco seed can perish in less than 4
hours under these conditions.

 Page 68
An excess of moisture during germination can be a major problem. A
moistened seed metabolizes carbohydrates and other seed reserves to
supply energy requirements for germination. A lack of oxygen caused
by excessive moisture reduces respiration, a critical step for energy
mobilization in the seed. Excess moisture provides an environment
suitable for certain seedling diseases. Seed sown in a properly
prepared, well drained seedbed seldom encounters an excess of water
in most production areas. In contrast, an overabundance of moisture is
one of the leading causes of poor germination and seedling failure in
float systems and other new transplant production methods.
c
Temperature
Tobacco seed is capable of germinating over a wide range of
temperatures, but the optimum temperature range is 18° to 23°C.
Deviation from this range slows germination rates and reduces the
percentage of seed that germinate successfully. Within this optimum
range, most seed will germinate within 7 to 12 days, but at 15°C,
germination may take an additional 1 to 2 weeks and it will occur
unevenly. Temporary cold periods, down to freezing or below, will not
destroy the seed unless the radicle is just emerging, but they will delay
emergence.
High temperatures in a greenhouse during germination are one of the
major causes of seed failure when using the newer transplant
production methods. Using unpublished data from an experiment
employing a graduated temperature table, we found that raising the
temperature to 26°C reduced germination rates 1 to 2%. Between 3
and 4% fewer seed germinated when temperatures were increased to
29°C, and at 32°C germination was reduced 8 to 9%. At temperatures
above 32°C, germination rates decrease to unacceptable levels. Some
varieties appear to tolerate higher temperatures during early growth
stages, but this tolerance is not as great during germination as it is
after seedling establishment.
Types of Seed
a
Raw Seed
Seed that is harvested, dried and packaged without any further
amendments or processing is classified as raw seed. This traditional
seed product, that is used to sow prepared soil beds, has been
marketed in packages weighing as little as a few grams to as large as a
kilogram or more. In the USA, tobacco grown from raw seed now
accounts for less than 25% of the total tobacco acreage. In this market,
raw tobacco seed that is sold as certified seed, which guarantees
genetic purity of the seed, must have a germination rate exceeding
80% and have little foreign matter or seed of other plant species. Raw
seed, however, is generally unsuitable for the newer transplant
production systems.
b
Pelleted Seed
The pelleting of tobacco seed has become necessary due to the growth
of container and precision-seeded transplant production. The
prevailing thinking is that the pelleting process, increasing seed size
by adding foreign material to the seed surface, is done only for one
reason to enable the mechanical handling of individual tobacco seed.
However, there are two other important reasons. Firstly, a pelleted
seed can be more accurately placed in the soil profile, and secondly, a
larger pelleted seed will remain positioned at a precise soil depth
better than a raw seed.
As described previously, proper moisture availability is critical to
uniform germination. The material forming the pellet can have an
important role in maintaining proper moisture around the germinating
seed. Indeed, the pellet can provide a moisture reserve for the seed
during variable environmental conditions. Gawande, et al. (1980)
found that pelleting may also provide protection against thermal stress
by reducing the rate of temperature change for the seed. There are
many other agronomic and horticultural seed that come in a pelleted
form, and the materials used in the pelleting process are just as varied.
The pelleting process and the material used to form the pellet can be
unique for each crop, and using materials or methods designed for
other crops to pellet tobacco seed can reduce or delay germination.
Thus, care should be taken to use only those processes proven to be
effective for tobacco.
c
Primed and Pelleted Seed
Seed of lettuce, tomato, pepper, onion and other crops have been
primed before they are marketed to growers for the past several years.
In priming, the seed are allowed to germinate under special
conditions, and then, the germination process is stopped at a precise
point. The seed is then pelleted prior to delivery to the growers. This
technique has been applied to tobacco seed, and primed seed has been
shown to germinate faster and maintain high germination rates at a
wider temperature range than seed that has not been primed.

 Page 69
Using primed seed can reduce needed heat levels in a greenhouse and
provide more uniform germination at relatively wide temperature
ranges.
Seed Storage
Tobacco seed, unlike seed of many other agronomic crops, can remain
viable for many years if it is stored under proper conditions. However,
the level of seed quality needed by growers can diminish to
unacceptable levels within several years following harvest if key
storage practices are not followed. The quality of the seed at the time
it is placed into storage is a major determinant of seed longevity. A
seed lot that has poor or substandard initial quality will deteriorate in
storage faster than seed with better germination and vigor when it is
first stored.
Obviously, the rate of seed decline during storage is a function of
storage conditions. In general, acceptable environmental conditions
for seed storage can be defined as:
%RH + °F < 100
where %RH is percent relative humidity and °F is the temperature in
degrees Fahrenheit. For example, the temperature in a storage room
kept at 40% relative humidity should be no greater than 60°F (16°C).
Under these conditions, large quantities of raw seed are best stored in
containers, such as cloth bags, that permit air exchange. This allows
for moisture and temperature equilibration throughout the seed lot. On
the other hand, pelleted seed are often kept in sealed containers to
prevent the coating material from absorbing moisture. The proper,
long-term storage conditions for primed seed may vary depending
upon the priming method and the coating method and material. Small
quantities of research or breeders' seed can be stored in sealed
containers kept in freezers at -10°C or less. The keys to successful,
long-term storage of tobacco seed in a freezer are low moisture
content (8% or less) in the seed and allowing the seed and its
container to equilibrate to room temperature before the container is
opened for sampling.
References
Gawande, M., Mohapatra, S.C. & Johnson, W.H. (1980) Effect of
seed size and pelletization on tobacco seed germination under varying
temperature regimes. Tob. Sci., 24, 4952.
Gwynn, G.R. (1973) Effect of maturity on germination of tobacco
seed of six tobacco cultivars. Tob. Sci., 17, 108109.
Kasperbauer, M.J. (1968) Germination of tobacco seed. I.
Inconsistency of light sensitivity. Tob. Sci., 12, 2022.
Litton, C.C. & Stokes, G.W. (1964) Outcrossing in burley tobacco.
Tob. Sci., 8, 11315.
Mohapatra, S.C. & Johnson, G.W. (1978) Development of the tobacco
seedling. 1. Relationship between moisture uptake and light sensitivity
during seed germination in a flue-cured variety. Tob. Res., 4, 419.
Spaulding, D.W. & Steffens, G.L. (1969) Elimination of light
requirements for tobacco seed germination with gibberellic acid,
indole-3-acetic acid and N6 benzyladenine. Tob. Sci., 13, 1569.
Thomas, J.F. & Raper, C.D., Jr (1979) Germinability of tobacco seed
as affected by culture of the mother plant. Agron. J., 71, 6945.

 Page 70

4B
Seedling Production
W.D. Smith
North Carolina State University
Raleigh, North Carolina, USA
Introduction
Traditionally, tobacco seedlings have been produced in plant beds
covered with perforated plastic or woven materials in temperate
regions or mulched with gravel, sand, rice hulls and other materials in
tropical regions. In recent years, labor shortages and the probable ban
on the use of methyl bromide for soil sterilization have led to the use
of containerized (intact-root) seedlings in many areas of the world.
The most common method for the production of intact-root seedlings
is the float system. The float system was introduced by Speedling, Inc,
a large producer of vegetable and other transplants in the USA, in the
mid 1980s. Since its introduction, the float system has become the
most common method of seedling production in the USA. For
example, approximately 73% of the tobacco seedlings in North
Carolina were produced in greenhouse float systems during 1996
(Smith & Boyette, 1997). Various adaptations of the float system are
also used in many Western European countries, Australia and Mexico
as well as countries in Central and South America.
In the float system, tobacco seedlings are grown in polystyrene trays
that contain a soilless growing medium. These trays are often called
Todd cells because they were developed by Mr George Todd, founder
of Speedling Inc. The trays are floated on a bed of water that has been
fertilized with a soluble fertilizer. Each tray contains 200392 cells that
are shaped like an inverted pyramid. In the flue-cured production
areas of the USA, the float plants are grown in heated greenhouses. In
the burley production regions, where production units are often
smaller than with flue-cured, small heated greenhouses or covered-
outdoor water beds are used. In tropical regions, conventional
greenhouses are not necessary for float systems.
Intact-root greenhouse seedling production has been examined for
several years as an alternative to field or greenhouse plant beds. In
Canada, seedlings produced in Todd cells were more uniform,
underwent less transplanting shock and survived better than bareroot
seedlings from conventional plant beds in greenhouses (Walker, 1980;
Walker, 1981; Lamarre, 1986). The proportion of root to shoot was
much higher for seedlings from Todd cells than for bareroot seedlings
(Walker, 1980). In the field, plants from Todd cells flowered earlier
than barefoot seedlings. Walker (1981) compared the field
performance of seedlings produced in two cell sizes, 28 cm3 and 36
cm3. Survival after transplanting depended more on field conditions
than on cell size. Days to flower, yield and cured leaf quality were
similar for both cell sizes.
Currently, cell numbers range from 200 to 392 cells per tray. Some
commonly used trays include the following cell numbers and volume
per cell:

Cells/trayCell volume (cm3)

200 27
242 23.5
253 16
288 17
392 13.6

Studies by Pfeiffer, et al. (1990), Suggs and Mohapatra (1988) and

Reed (1996) showed similar field performances among plant bed
versus intact-root seedlings and among intact-root seedlings grown in
different cell sizes. Pfeiffer, et al. (1990) reported that intact-root
seedlings flowered 5 days later than bareroot seedlings, but flowering
was more uniform. In addition, less premature flowering was
observed in greenhouse seedlings than with bareroot seedlings.
Economic studies have shown greenhouse float systems to be slightly
more expensive than field plant beds (Lychak & Brown, 1995).
However, float systems offer significant labor advantages as
compared to plant beds. In particular, the float system saves labor
during the critical planting period, allowing tobacco to be transplanted
in a timely manner.

 Page 71
Seedling Production in the Float System
a
Water Quality
Water quality factors that have been reported to impact seedling
production include high bicarbonate (Smith, et al., 1993; Rideout, et
al., 1995), low and high boron concentrations (Rideout & Gooden,
1996; Smith & Boyette, 1997) and low calcium (Rideout & Gooden,
1996). General guidelines for the nutritional suitability of water
sources are shown in Table 4.1.
Table 4.1 Desirable characteristics for water sources
used in seedling production with the float system.
Component Range Component Range
N03-N (mg/L) 0-5 SAR 04
P (mg/L) 05 EC (mS/cm) 0750
K (mg/L) 010 TC (meq/L) 02
Ca (mg/L) 20100 Cl (mg/L) 070
Mg (mg/L) 625 Na (mg/L) 070
S (mg/L) 025 Al (mg/L) 05
Fe (mg/L) 02 F (mg/L) 01
Mn (mg/L) 02
Zn (mg/L) 02
Cu (mg/L) 02
B (mg/L) 02
Mo (mg/L) 00.1
Source: Smith, et al. (1993).

Bicarbonate levels are highly variable among water sources within a

region and among regions. In general, bicarbonate levels above 2
meq/l require neutralization with acid (Coggins, 1993; Rideout, et al.,
1995). Bicarbonate levels greater than 2 meq/l can be tolerated in the
presence of high levels of calcium (R.C. Pearce, Department of
Agronomy, University of Kentucky, personal communication). In the
USA, sulfuric acid (H2SO4) is the most commonly recommended
acid. Nutritional work by Rideout and Gooden (1996) has shown that
excessive phosphorous concentrations in float water increased
seedling stem length and decreased stem diameter. Therefore,
phosphoric acid (H2PO4) (commonly used in horticultural systems) is
less desirable than H2SO4 for tobacco seedling production.
b
Growing Media
In the USA, tobacco media are peat-based with various combinations
of vermiculite and perlite. Particle size distribution and nutrient
charge are important factors in the suitability of a medium for tobacco
transplant production. Particle size in a soilless medium is similar to
the texture of a soil and is determined by the relative amounts and size
of the components in the medium (Fonteno, 1993). The particle size
distribution of a medium determines many characteristics that are
important in plant growth, such as aeration, water holding capacity,
drainage and capillarity (Tilt, et al., 1987).
Research by Harrell (1995) indicated that a wide range of media
particle sizes were suitable for tobacco seedling production. Very
coarse-textured media, such as those with 500% or more perlite,
promoted dry cells and those with 100% peat were less satisfactory
than media that contained perlite and/or vermiculite. Proper filling of
the trays with the growing medium is important in preventing
oversaturation of the medium. Trays should be loosely filled to avoid
compaction. Medium saturation with water, resulting from an overly-
compacted medium, increased the incidence of negative geotropism
and decreased root mass of float plants in a study by Cui (1994).
Research by Harrell (1995) indicated that successful transplant
production can be obtained with noncharged (lime and CaSO4 only,
no additional fertilizer added) medium when all of the nutrients were
applied through the waterbed. This research showed that fertilizer
salts accumulated in the medium from the upper 1.25 cm of the tray.
High temperatures, low humidity and excessive air movement
promote water evaporation from the surface of the growing medium,
which results in the accumulation of fertilizer salts in the upper
portion of the cell. Salts can accumulate to levels high enough to
injure seedlings, even when recommended fertilization programs are
followed. If high fertilizer salts levels are detected during the first 4
weeks after seeding (> 1000 microseimens in the medium from the
upper 1.25 cm of the cell), water should be applied uniformly from
over-top to reduce fertilizer salts levels (Smith & Boyette, 1997).
c
Nutrition
Jones, et al. (1992) reported successful seedling production in a float
system with a two-step fertilization program. This program included
an application to the float water of 150 mg nitrogen/liter (N/L) from
201020 (N-P2O5-K2O) fertilizer at seeding followed by a second
application of 100 mg N/L from ammonium nitrate (NH4NO3) 4
weeks later. More recently, Rideout and Gooden (1996) reported a
positive benefit

 Page 72
from limiting the phosphorous levels in water to no more than 40 mg
phosphorus pentoxide/liter (P2O5/ L). Seedlings grown at lower
phosphorous levels grew slower, required fewer clippings and had
larger stem diameters than those grown with water levels exceeding
40 mg P2O5/L. This program has been modified for burley seedling
production to include a recommendation of no more than 100 mg N/L
based on the observance of excessive stem rot diseases (primarily
from Erwinia spp.) under the higher nitrogen regimes recommended
for flue-cured seedlings.
It appears that the form of nitrogen is important in seedling growth in
float systems. Research conducted in 1994 showed reduced seedling
growth when more than half of the total nitrogen in a fertilizer was
provided from urea, as compared to growth obtained from a fertilizer
with all of the nitrogen supplied as nitrate and ammonium (Table 4.2).
Similar results have been observed in Kentucky (R. C. Pearce,
Department of Agronomy, University of Kentucky, personal
communication). The observed reductions in plant growth may be a
result of nitrite toxicity.
Table 4.2 Effect of urea concentration in the
fertilizer on seedling stem length, fresh weight
and dry weight.
Urea Stem Fresh Dry
concentration length weight weight
(% of total (cm) (g/20 seedlings)
N)
0 5.4a 77a 5.3a
52 3.8b 54b 4.0b
77 4.4b 43c 2.9c
Note: treatment values within a column
followed by the same letter are not statistically
different and should be considered similar.
Source: Smith & Boyette (1997).
Sulfur deficiency has been reported in float systems if the medium
was not supplemented with MgSO4 or CaSO4 (Smith & Boyette,
1997). The addition of 2530 mg S/L from MgSO4 has successfully
corrected deficiencies.
Boron deficiency has been reported in North Carolina, South
Carolina, Tennessee and Kentucky. In most cases, the water did not
contain measurable boron, and the fertilizer also did not contain
boron. General recommendations from state extension services are for
the selection of soluble fertilizers that contain boron at concentrations
that will supply 1.02.0 mg B/L when used at a rate to supply 150 mg
N/L (Rideout & Gooden, 1996; Smith & Boyette, 1997). General
recommendations are to select a soluble fertilizer with a complete
micronutrient charge.
d
Clipping
Proper clipping is an important tool in increasing the number of
usable seedlings, transplant hardiness, stem length and diameter
uniformity. A properly clipped plant is essential for carousel
transplanters, because uniform stem lengths are important for the
machine to transplant seedlings at the proper depth and excessive
foliage disturbs the timing mechanism. Clipping can be used to delay
transplanting when field conditions are unfavorable. Past research has
shown that maximum usability is obtained with three to five clippings
(R.C. Long, Department of Crop Science, North Carolina State
University, personal communication).
While proper clipping is a significant benefit, recent research has
shown that improper clipping can adversely affect stem length,
increase stem rots and slow plant growth in the field. Research
conducted in 1995 and 1996 by Reed (1996) in Virginia showed that
the time of initial clipping and the severity of clipping is important in
affecting seedling stem length. For example, severe clipping
decreased stem length but did not increase stem diameter, as
compared to normal clipping. This research also showed that early
clipping also reduced stem length. Additional research in North
Carolina has shown that severe clipping, down to the bud,
immediately before transplanting reduced early-season growth in the
field and delayed flowering (Fisher, 1997). Current recommendations,
based on the work of Reed (1996), are to begin clipping at 3 to 5 day
intervals when plant height is 5 to 7 cm to the apical bud, and to set
the blade height at 2.5 to 3.5 cm above the apical bud.
Transplant Production in Plant Beds
Proper selection of the plant bed site is the first step toward producing
quality seedlings that are available for transplanting on a reasonably
predictable date. According to Collins and Hawks (1993), a good
plant bed location, particularly in temperate regions, should have:
a deep, fertile, well drained soil that warms up quickly;
soils with a 5% slope to the south, which are warmer than those with
slopes oriented in other directions;

 Page 73
windbreaks to the north and west, which protect plant beds from cold
temperatures and drying winds;
exposure to sunshine from 9 AM to 3 PM; and
a clean water supply for irrigation.
a
Chemical Alternatives to Methyl Bromide for Soil Fumigation
Methyl bromide has been a common fumigant in tobacco plant beds
since its introduction in the 1950s (Collins & Hawks, 1993). It is a
very effective and convenient soil fumigant. However, concern over
the potential negative impact of methyl bromide on the stratospheric
ozone layer has led to a planned worldwide suspension of methyl
bromide use on 1 January 2001 (Ferguson & Padula, 1994).
With this potential loss of methyl bromide, recent research has
focused on evaluating a number of chemical alternatives. Miner and
Worsham (1990) reported that dazomet provided similar weed control
to methyl bromide. Csinos (1995), working in Georgia, has reported
excellent control of weeds, nematodes and soil pathogens with a
mixture of metam sodium and Telone II. This combination seems to
provide the best alternative to methyl bromide for both efficacy and
seedling phytotoxicity.
b
Seeding
The rate of seeding not only determines the number of seedlings in a
plant bed, it also determines seedling quality. Excessive seeding rates
result in reduced stem diameter, slower growth and poor survivability
in the field (Dean, et al., 1960; Jones & Terrill, 1984). Lower than
optimum seeding rates often result in 'bunchy' seedlings that have
short stems and large leaves. Seltmann (Collins & Hawks, 1993)
reported that these bunchy seedlings were more likely to flower
prematurely and to produce lower yields than optimum sized
seedlings. Dean, et al. (1960) reported that one seedling per 5 cm × 5
cm plant bed area provided the best quality seedling. Therefore, a
seeding rate of 0.5 gm per 10 m2 should be adequate.
It is generally believed that a good quality seedling for environmental
conditions in the southeastern USA has a stem length of 7.513.5 cm
and a stem diameter of approximately 8 to 10 mm (Collins & Hawks,
1993). However, research in Zimbabwe has shown an advantage of
larger seedlings, 15 to 17 cm in length, for dry-season stand
establishment (Tobacco Research Board, 1990).
Since tobacco seeds are so small, they are usually mixed with other
materials such as sand, fertilizer, lime or water during the seeding
operation. In recent years in the USA, belt seeders have been used to
sow pelleted seeds. Studies conducted by Smith (1991) compared four
seeding methods (conventional, belt seeder, water and a drop
spreader) that are available to farmers in the USA. Plant stands were
similar among the seeding methods. However, methods such as the
belt seeder that provided more uniform stands resulted in reduced
pulling times, and thus reduced labor requirements per transplanted
hectare.
c
Nutrition
Fertilization levels will vary a great deal depending on soil type,
rainfall and temperature. Collins and Hawks (1993) suggested the
following for flue-cured seedlings on soils in the southeastern USA:
1.6 kg N, 0.8 kg P2O5, 0.8 kg K2O and 0.6 kg MgO per 10 m2 plant
bed. No more than 2.3 kg N, 2.3 kg K2O and 2.3 kg P2O5 per 10 m2
are recommended for burley tobacco plant beds (Palmer, et al., 1993).
Dean, et al. (1960) reported better seedling growth prior to and after
transplanting when seedlings were fed all NO3-N as compared to
plants that received all nitrate nitrogen (NO3-N) for 40 days followed
by all ammonium (NH4-N) from (NH4)2SO4. Therefore, the use of
high NH4-N sources is discouraged.
Excessive nitrogen rates are also detrimental to seedling growth in the
field. Dean, et al. (1960) reported that as nitrogen levels increased to
excessive levels, root number and root mass 7 days after transplanting
decreased.
d
Clipping
Clipping was first reported in Zimbabwe as a way to regulate seedling
growth prior to transplanting (Kille, 1970). Papenfus (1978) reported
an added benefit of increased stem length uniformity. More recently,
clipping was investigated as a way to increase stem length uniformity
to make once-over mechanical seedling removal practical. Studies by
Cundiff and Miles (1980), Miner, et al. (1983a) and Stephenson, et al.
(1984) showed that clipping increased the number of seedlings
available in one pulling from 1020% of the total seedlings in the bed
to over 50%. This increase was not enough for mechanical pulling but
it did allow for a reduction in the plant bed area per planted hectare by
20 to 30%. Increased uniformity

 Page 74
also has been shown to increase the efficiency of seedling removal
from the plant bed by hand (Miner, et al., 1983a; Stephenson, et al.,
1984). Miner, et al. (1983a) reported maximum usability was obtained
with two to three clippings.
Several researchers also reported that clipping could be utilized to
delay transplanting when field conditions were not suitable, and that
clipping was better than the more traditional method of pulling and
storing seedlings (Cundiff & Miles, 1980; Miner, et al., 1983a; Jones
& Terrill, 1984). Clipping seedlings 1.32.5 cm above the bud when
the plants were approximately 10 cm in height to the bud followed by
a second clipping 5 days later delayed transplanting by 710 days.
The field performance of clipped seedlings has been reported by a
number of researchers to be similar to nonclipped seedlings (Miner, et
al., 1983b; Jones & Terrill, 1984; Suggs, et al., 1988; Pfeiffer, et al.,
1990). Clipping is now a standard practice in many areas of the world
whether it be for hardening, delaying transplanting or increasing
production efficiency. In 1996, 88% of the plant beds in North
Carolina were clipped (Smith & Boyette, 1997).
e
Undercutting
This practice was initially examined as a way to harden seedlings for
transplanting. For example, undercutting to prune root systems was
examined in Zimbabwe (Tobacco Research Board, 1985, 1987), and
Walker and Reynolds (1982) investigated the 'forking' of seedlings in
muck beds in Canadian greenhouses prior to transplanting.
Dean, et al. (1960) reported that seedlings subjected to hardening
treatments that consisted of moisture stress in combination with cool
temperatures grew more rapidly, had higher carbohydrate levels and
had greater root development after transplanting than nonhardened
seedlings. Some 22 years later, Walker and Reynolds (1982) reported
that seedlings that were forked had larger root systems prior to
transplanting and that the forked plants survived better in the field. In
addition, the forked plants outyielded nonforked plants 2 of the 3
years in the study. Research in Zimbabwe showed that root pruning
from undercutting increased root volume and starch concentrations in
seedlings before and during the first 6 weeks after transplanting
(Tobacco Research Board, 1985, 1987). However, yield and quality
were unaffected. Suggs, et al. (1988) reported that undercut plants
flowered about 5 days earlier than nonundercut seedlings. However;
there were no differences in survivability, growth rate and yield due to
undercutting treatment.
Undercutting, without root pruning, has been examined as a pulling
aid to increase the efficiency of seedling removal from plant beds.
Smith (1991) reported an increased pulling efficiency of
approximately 400 seedlings per man hour with undercutting
immediately prior to pulling as compared to nonundercut plant beds at
two locations in 1990.
References
Coggins, T.E. (1993) Effect of sodium and bicarbonate on tobacco
seedling production in the greenhouse float system. Unpublished MS
thesis, North Carolina State University.
Collins, W.K. & Hawks, S.N., Jr (1993) Principles of Flue-Cured
Tobacco Production, 1st ed. NC State University, Raleigh, NC 27695.
Csinos, A.S. (1995) Evaluation of alternative materials for methyl
bromide soil fumigation. 1995 Georgia Tobacco Research-Extension
Report. University of Georgia.
Cui, M. (1994) Seed germination and seedling development of burley
tobacco in a greenhouse float system. Unpublished MS thesis,
University of Kentucky, Lexington.
Cundiff, J.S. & Miles, J.D. (1980) Effect of transplant clipping on
stem length, variability, and field production. Tob. Sci., 24, 14.
Dean, C.E., Seltmann, H. & Woltz, W.G. (1960) Some factors
affecting root development by transplanted tobacco transplants. Tob.
Sci., 4, 1925.
Ferguson, W. & Padula, A. (1994) Economic effects of banning
methyl bromide for soil fumigation. USDA-ERS, Economic Report No
677.
Fisher, L.R. (1997) Factors affecting early field growth of flue-cured
tobacco transplants from the float system. Unpublished MS thesis,
North Carolina State University.
Fonteno, W.C. (1993) Problems and considerations in determining
physical properties of horticultural substrates. Acta Hortic., 342,
197204.
Harrell, N.E. (1995) The effects of physical properties of soilless
media on tobacco seedling production in the float system.
Unpublished MS thesis, North Carolina State University.
Jones, J.L. & Terrill, T.R. (1984) Effects of transplant size and
condition on survival, yield, and quality of flue-cured tobacco. Tob.
Sci., 28, 737.
Jones, M.A., Miner, G.S. & Smith, W.D. (1992) Effects of media and
fertilization on the direct seeded float system. Tob. Sci., 37, 1317.
Kille, T. (1970) Early preparations for this year's tobacco crop. Rhod.
Tob. J., 22(9), 257.
Lamarre, M. (1986) Field evaluation of tobacco plants produced in
Todd cells under various conditions. The Lighter, 56(4), 516.
Lychak, T. & Brown, A.B. (1995) Producing tobacco transplants in
greenhouses: production costs. NC Coop. Ext. Serv. Bull. AG 4884.


 Page 75
Miner, G.S., Cundiff, J.S. & Miles, J.D. (1983a) Effects of clipping
flue-cured plantbeds on transplant production efficiency and
uniformity. Tob. Sci., 27, 7074.
Miner, G.S., Cundiff, J.S. & Miles, J.D. (1983b) Effects of clipping
flue-cured tobacco plantbeds on field performance of clipped plants.
Tob. Sci., 27, 756.
Miner, G.S. & Worsham, A.D. (1990) Fumigation of tobacco
plantbeds with dazomet. Tob. Sci., 34, 827.
Palmer, G., Maksymowicz, B. & Calvert, J. (1993) Transplant
production. In: Tobacco in Kentucky. Kentucky Coop. Ext. Ser. Bull.
ID 73.
Papenfus, H. (1978) Seedling uniformity and growth. Rhod. Tob.
Today, 1(13), 213.
Pfeiffer, I., Smith, W.D., Collins, W.K., et al. (1990) Field
performance of clipped, nonclipped, and intact-root flue-cured
tobacco seedlings. Tob. Sci., 34, 258.
Reed, T.D. (1996) Float greenhouse tobacco transplant production
guide. Va Coop. Ext. Serv. Bull. 43651.
Rideout, J.W. & Gooden, D.T. (1996) Greenhouse seedling production
recommendations. In: South Carolina Tobacco Growers Guide 1997.
Clem. U. Coop. Ext. Serv. Bull. EC-569.
Rideout, J.W., Gooden, D.T. & Martin, S.B. (1995) Corrective
measures for growing tobacco seedlings using the float system with
water high in bicarbonate. Tob. Sci., 39, 13036.
Smith, W.D. (1991) Transplant Production. In: 1991 Flue-Cured
Tobacco Information. NC Coop. Ext. Serv. Bull. AG-187.
Smith, W.D. & Boyette, M.D. (1997) Transplant production. In: 1997
Flue-Cured Tobacco Information. NC Coop. Ext. Service Bull. AG-
187.
Smith, W.D., Peedin, G.F., Yelverton, F.H. & Cambell, C.R. (1993)
Producing tobacco transplants in greenhouses: water quality. NC
Coop. Ext. Serv. Bull. AG 4883.
Stephenson, M.G., Miles, J.D., Gaines, T.P. & Wilson, W.H., Jr (1984)
Clipping effects on transplant yield and field performance of flue-
cured tobacco. Tob. Sci., 28, 558.
Suggs, C.W. & Mohapatra, S.C. (1988) Tobacco transplants 2. Effects
of bare-root versus intact-root plants on yield, value, growth rate, and
chemistry. Tob. Sci., 32, 16.
Suggs, C.W., Mohapatra, S.C. & Johnson, W.H. (1988) Tobacco
transplants. Part 3. Effect of clipping and undercutting on yield, value,
chemistry, and growth. Tob. Sci., 32, 248.
Tilt, K.M., Bilderback, T.E. & Fonteno, W.C. (1987) Particle size and
container size effects on growth of three ornamental species. J. Am.
Soc. Hort. Sci., 112(6), 9814.
Tobacco Research Board (1985) Root development. In: Annual
Research Report 1. pp. 81223. Harare.
Tobacco Research Board (1987) Root pruning. In: Annual Research
Report 2. pp. 3745. Harare.
Tobacco Research Board (1990) Section A production of seedlings.
In: Flue-Cured Recommendations. Harare.
Walker, E.K. (1980) Culture of flue-cured tobacco transplants in Todd
cells. The Lighter, 50(4), 1222.
Walker, E.K. (1981) Culture of flue-cured tobacco seedlings in Todd
cells: influence of size of cell, age of seedlings, and time of field
transplanting. Tob. Sci., 25, 97101.
Walker, E.K. and Reynolds, L.B. (1982) Greenhouse culture of flue-
cured tobacco seedlings: Influence of media and forking seedbeds.
Tob. Sci., 27, 7781.

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4C
Field Practices
K.C. Flower
Tobacco Research Board
Harare, Zimbabwe
Introduction
This section deals with the effect of various practices in the field on
the growth, yield, quality and chemical composition of tobacco. The
physiological basis of these factors is discussed, rather than the
practical aspects, as this is applicable to most tobacco types. Cultural
practices from transplanting through to topping are covered in the
sections on establishment, plant populations, nutrition and topping and
suckering. Environmental factors, particularly rainfall, have a major
effect on the growth and ripening of tobacco and this is discussed in
the sections on plantwater relations and plant response to climatic
factors.
Establishment
It is essential that good quality seedlings are used as transplants. They
should be about 15 cm in height and free from any pests or diseases.
Increasing seedling size markedly increases dry-matter production
until just before topping, after which the effect diminishes (Tobacco
Research Board, 1981). Transplants should not be too large as the
number of harvestable leaves may be reduced as a result of early
flower initiation (McCants & Woltz, 1967). It is important that most
of the root system is retained at pulling and the entire plant maintained
in a turgid condition after pulling to transplanting. Seedlings need to
be placed into moist soil in the field with the terminal bud at least 2 to
3 cm above ground level.
When planting under hot and dry conditions, seedlings should be
hardened by withholding water at least 14 days before planting
(Tobacco Research Board, 1990a). A seedling is considered to be
'hard' when the stem does not break if it is either bent double or
wrapped around the finger (Akehurst, 1981b). Hardened seedlings
have more starch in the stem and roots and a greater dry mass and root
volume than non-hardened ('soft') seedlings (Tobacco Research
Board, 1986a), although excessive hardening may result in a lower
starch concentration (Tobacco Research Board, 1983). Dean, et al.
(1960) demonstrated a relationship between the level of soluble
carbon in transplants and root development after transplanting.
Excessive nitrogen applied in the seedbeds increases the total nitrogen
and decreases the starch and sugar concentrations of seedlings. Such
seedlings are difficult to harden (Tobacco Research Board, 1987a;
Tobacco Research Board, 1988a) and their ability to survive drought
conditions is poor. In another study, strong positive correlations were
found between starch (r2 = 0.90; p < 0.01) and sugar (r2 = 0.70; p <
0.05) concentrations at transplanting and plant growth rate/day in the
first three weeks after transplanting (Ryding & Wixley, 1984).
Therefore, hardened seedlings tend to grow faster initially and can
survive drought conditions better than soft seedlings. Nonetheless,
provided there is adequate soil moisture for growth, soft seedlings
perform as well as hard ones (Walker, 1974; Tobacco Research Board,
1981). Under irrigated conditions, soft and hard seedlings also had
similar yields, but the soft seedlings had higher grade indices and
lower total alkaloid values in the bottom leaves (Tobacco Research
Board, 1986a).
Removal of a portion of the root system (root pruning) prior to or in
the middle of hardening significantly increases the root volume, starch
concentration and early growth of transplants in the field. Root
pruning involves pulling a blade or wire attached to two ripper tynes
under the seedbed at a depth of about 10 cm. The beds must be
watered before and after this operation (Tobacco Research Board,
1990a). The enhanced growth effect is not measurable by about 6
weeks after transplanting, and the yield and grade index of cured
leaves are not affected (Tobacco Research Board, 1985a; Tobacco
Research Board, 1987b). Consequently, root pruning is mainly used to
prevent overgrowth in seedbeds. Root pruning should not be confused
with undercutting, where the objective is to loosen seedlings in order
to ease pulling (Smith & Boyette, 1995).
Clipping of seedlings is generally done to improve

 Page 77
uniformity (Stephenson, et al., 1984; Smith & Boyette, 1995). There
is evidence that this practice reduces the initial growth rate in the field
(Tobacco Research Board, 1985a; Tobacco Research Board, 1986b);
however, this was not confirmed by Suggs, et al. (1988). Under
climatic conditions that were suitable for premature flowering, i.e.
short daylength and/or low temperatures, the removal of portions of
leaves by clipping significantly increased the number of harvestable
leaves and yield of tobacco, although the optimum time to clip varied
(Tobacco Research Board, 1985a; Tobacco Research Board, 1986c;
Tobacco Research Board, 1987c; Pfeiffer, et al., 1990). However,
Miner, et al. (1983) reported that clipping had no effect on yield,
grade index, alkaloids and reducing sugar concentrations. Papenfus
(1980) reported that, under short days (8 hours) and cool nights
(27/15°C, day/night), five to seven leaves/plant (500 cm2 of leaf area)
were required for floral initiation of the cultivar Kutsaga El, while in
another study (Tobacco Research Board, 1987c) the same cultivar was
most susceptible at the 8 to 12 leaf stage. Hopkinson (1969) found
that three leaves were sufficient for the induction of flowering in a
mammoth mutant. These results suggest that it is difficult to define the
optimum time for clipping to reduce premature flowering, and that it
is related to the cultivar, stage of growth or leaf area of the seedlings
and the severity of the inductive conditions.
The degree of hardening, size of seedlings and amount of water
required for good crop establishment varies according to the climatic
conditions prevailing before and after transplanting and the
availability of water for irrigation. Where conditions are hot and dry,
as for the early season crop in Zimbabwe, seedlings should have a
stem 15 to 17 cm long and 6 to 10 mm thick and they should be
hardened for a period of 2 to 4 weeks (Tobacco Research Board,
1990a). Sufficient water is added to the planting hole so that it drains
to the subsoil (residual) moisture. Seedlings are placed at a suitable
depth in the moist soil to prevent the roots from desiccating and the
stem from burning on the hot ground. Where conditions are mild and
moist, or where soil moisture is frequently replenished, smaller
seedlings of 8 to 10 cm can be used (Papenfus & Quin, 1984). Suggs
and Mohapatra (1987) concluded that there were no differences in
yield, value, growth rate and sugar and alkaloid concentrations
between seedlings of 13 to 30 cm in height. However, very small
seedlings result in significantly lower yields (Tobacco Research
Board, 1981).
In recent years there has been a large increase in the production of
seedlings which have intact roots when transplanted into the field.
These seedlings are normally produced in greenhouses using
polystyrene trays or pots, and they are generally smaller than those
from seedbeds. Studies have shown that the growth rate of plants with
intact roots is often greater than that of bare-root seedlings during the
first weeks after transplanting. However, there are no differences in
yield, grade index or reducing sugar and total alkaloid concentrations
(Suggs & Mohapatra, 1988; Pfeiffer, et al., 1990).
Under favorable conditions, new roots are formed 4 days after
transplanting, but no measurable increase occurs in the dry weight of
the above ground parts until about 10 days later, and only small
increases occur during the subsequent 7 to 10 days. The period of
major increase in dry weight is normally from the fourth to eighth
week after transplanting. With the onset of flowering, which occurs 7
to 8 weeks after transplanting, there is a sharp decrease in the rate of
dry matter production (McCants & Woltz, 1967).
Plant Populations
Many studies have shown that the population of tobacco plants has a
great influence on their growth, yield, quality and chemical
composition. Furthermore, it should be realized that factors such as
climate, planting date, method and rate of fertilization, time and
height of topping and missing plants can affect the results. Tobacco
has been considered as a source of protein for food, synthetic smoking
materials and various bioengineered products (Tso & Gori, 1976;
Cole, 1994). This necessitated the investigation of systems of growing
flue-cured tobacco at populations of between 45 000 and 100 000
plants/ha, in order to maximize dry mass and protein production
(Campbell, et al., 1980; Woodlief, et al., 1981; Long, 1984; Tobacco
Research Board, 1995b).
There is a large variation in plant population among the different
tobacco types: dark air-cured and fire-cured tobacco have the lowest
populations of about 8900 plants/ha and Oriental the highest
population of about 148 260 plants/ha (Tso, 1990a), whereas flue-
cured and burley have populations of about 15 000 plants/ha. The
optimum plant population is often determined by leaf usability, or
quality, rather than yield. However, if dry conditions could be
predicted then plant populations could be decreased in order to reduce
water stress.
Plants should be spaced and managed to obtain a complete canopy to
make maximum use of sunlight. Excessively high populations must be
avoided because

 Page 78
of possible adverse effects on leaf quality caused by shading
(Papenfus, 1984). Andersen, et al. (1985) reported that the cured
leaves of burley tobacco that were grown under increased shade had
more red and less yellow hue, less brightness, higher chlorophyll,
carotenoid and nitrate-N concentrations and lower brown pigment and
total alkaloid concentrations. Chen and Huang (1970) found that the
rate of change in leaf mass/unit area of flue-cured tobacco was
positively related to light intensity.
The distance between rows of tobacco is normally set by the machines
that are used for the different cultural and harvesting operations.
Therefore, the required plant populations are most often achieved by
altering the spacing within the row, which has a greater effect on
growth, yield and quality than changing the spacing between rows
(Boyce, 1965; Papenfus, 1984). This was supported by the work of
Shaw and Wixley (1981), which showed that most compensation for
missing plants occurred in plants next to the gap, although there was a
small contribution from plants growing opposite a gap of two or more
spaces in the adjacent row. They found that replacing plants markedly
reduced the magnitude of the loss and that the negative effects of
missing plants increased with time between planting and replanting.
Original plants on either side of replaced plants had a greater leaf area
and dry mass/unit area than that of the average full-stand plant. From
this work, a model was described to predict the yield of plants in any
configuration that included missing plants (Wixley & Shaw, 1981).
a
Leaf Characteristics
Closer spacing of plants results in decreased leaf area and weight per
unit area, which is often referred to as body or thickness, and filling
value (Elliot, 1970a; Papenfus & Quin, 1984). Middle and upper
leaves are affected the most. Oriental tobacco leaves must be small
and this is achieved with intra-row spacings of 10 to 15 cm on soil of
poor fertility, whereas flue-cured and burley tobacco have larger
leaves and are grown with intra-row spacings of 50 to 60 cm. Dark
fire-cured tobacco has large and heavy bodied leaves and is grown
with intra-row spacings of 75 to 100 cm.
b
Yield and Quality
It is obvious that the number of leaves per ha is dependent on the plant
population and the number of leaves per plant. Woltz and Mason
(1966) found that, within certain limits, yield was closely correlated to
leaf population, and they and Collins, et al. (1969) concluded that 296
520 leaves per ha was optimal for flue-cured tobacco.
Cousins (1966) found that yield and grade index of Oriental tobacco
increased when the population increased from 71 660 to 287 031
plants/ha. However, the optimum population was 148 260 plants/ha
because of the practical problems associated with harvesting and
curing at the higher populations. When the flue-cured tobacco plant
populations were varied from 7507 to 25 253 plants/ha by altering the
intra-row spacing from 111 to 33 cm in rows 120 cm apart (Tobacco
Research Board, 1979; Papenfus, 1984; Papenfus & Quin, 1984), total
yield increased from 2442 to 3710 kg/ha from the lowest to the
highest population. The largest increase occurred from 7500 to 15 000
plants/ha, after which the increments became progressively smaller.
Although the proportion of nondescript and unsaleable leaf increased
with population, it represented less than 4% of total yield, and the
saleable yield followed total yield. Yield and leaf area/plant decreased
with increased population. The grade index of the bottom leaves
decreased with increased population, whereas that of the middle and
top leaves increased, except at the highest populations. As a result,
crop value increased markedly with increasing populations up to 15
000 plants/ha, and then tended to remain constant. Also, the
proportion of lemon grades increased with the closer spacings up to a
population of about 12 500 plants/ha, whereas the proportion of
primings, lugs and cutters increased up to 15 200 plants/ha. Further
decreases in spacing had relatively little effect on leaf quality.
c
Chemical Composition
When all of the fertilizer is applied uniformly to the entire area,
increased plant populations generally result in lower total alkaloid and
nitrogen concentrations (Wolf, 1962; Weybrew & Woltz, 1975;
Campbell, et al., 1982; Schlotzhauer, et al., 1989). However, if all the
fertilizer or side-dressings of nitrogen were applied near each plant,
the nicotine and total nitrogen concentrations often increased with
increased population (Tobacco Research Board, 1984a; Tobacco
Research Board, 1990b). The application of fertilizer nearer the plant
possibly compensated for the greater nutrient competition at higher
populations.
Papenfus (1984) reported that changes in population of flue-cured
tobacco had little effect on nicotine concentration of the lower leaves,
but if the population were increased the nicotine decreased in the
middle

 Page 79
and upper leaves. The reducing sugar concentration was greater with
increased plant populations in all leaves except those at the bottom of
the plant where sugars decreased. The sugar to nicotine ratio for the
whole plant increased from 4.0:1 at the lowest population of 7500
plants/ha to 6.2:1 at 15 000 plants/ha, but was little affected thereafter.
In another study involving three plant populations and three topping
heights, the total nitrogen concentration decreased and reducing sugar
concentration increased with increased population, and the values
were similar at a constant leaf number per ha. Also, nicotine
decreased with increased plant population, but higher topping was
more effective in decreasing the nicotine concentration than more
plants (Weybrew & Woltz, 1975). In their work on close-grown
tobacco, Campbell, et al. (1980) found that increasing the population
from 45 448 to 136 344 plants/ha decreased the total alkaloid
concentration and had no significant effect on the reducing sugar
concentration. However, the values of both these components were
significantly lower than those expected of conventionally spaced flue-
cured tobacco. Few reports are available on the quality of smoke from
tobacco grown at different populations.
In a study on burley tobacco, Schlotzhauer, et al. (1989) found that the
solanesol concentration was lowest and the chlorogenic acid highest
in leaves from high population plants. Pyrolysis studies indicated that
for high population plants, smoke tar and nicotine concentration
decreased and the tar to nicotine ratio increased when compared to
normal population plants.
Nutrition
The rate of growth and quantities of nutrients taken up vary
considerably and depend on the type of tobacco, soil type and fertility,
management and cultural practices, and the environmental conditions.
Therefore, it can be expected that values of nutrient uptake reported in
the literature reflect the local conditions under which the crops were
grown. Generally, dark fire-cured and burley tobacco have a high
nitrogen requirement and are mainly grown on fertile, medium-to
heavy-textured soils; flue-cured tobacco requires less fertile soils and
is grown on sands and sandy loams; while Oriental tobacco requires
soils which are low in nitrogen and so infertile sands are the preferred
soils.
This section concentrates on the main nutrients and their effect on the
growth, yield, quality and chemical composition of tobacco. Many
more nutrients than mentioned here affect tobacco growth and reviews
have been done by Elliot (1975a), Miner & Sims (1983) and Tso
(1990b and 1990c). Also, nutrient accumulation and plant analysis as
aids in fertilizing flue-cured and burley tobaccos were discussed by
Miner and Tucker (1990).
The total accumulation of dry matter by tobacco from time of
transplanting in the field until final harvest is characterized by a
sigmoid curve (Fig. 4.1). Raper and McCants (1966) reported that in
flue-cured tobacco, nitrogen and potassium were absorbed relatively
early in the growing season, whereas the absorption of phosphorus,
magnesium and calcium occurred at a relatively constant rate
throughout the entire growing season (Fig. 4.1). In contrast, Atkinson,
et al. (1977), Bruns and McIntosh (1988) and Bertinuson, et al. (1970)
showed that burley, Maryland and shade-grown cigar wrapper
tobaccos, respectively, continued to accumulate nitrogen and
potassium during the latter part of the growing season, mainly because
they are grown on more fertile soils. The absorption pattern of
phosphorus, magnesium and calcium in the shade-grown tobacco was
similar to that of flue-cured tobacco (Fig. 4.2).
 Fig. 4.1
Yield of dry matter and nutrients in the above ground parts of flue-cured
tobacco (derived from Raper & McCants, 1966).
a
Nitrogen
Although potassium is absorbed in the greatest quantity, nitrogen is
the key nutrient in tobacco fertilization. From the seedling stage
through to final harvest, the soil nitrogen regime affects the process of
plant

 Page 80

Fig. 4.2
Yield of dry matter and nutrients of shade-grown cigar wrapper tobacco
(derived from Bertinuson, et al., 1970).
development more than any other element (McCants & Woltz, 1967).
The Balance between Nitrogen and Carbohydrate Metabolism
Weybrew (1979) was convinced that it was the interplay of the
nitrogen and carbohydrate metabolism, as influenced by management
and weather, that predetermined the quality and chemical composition
of cured tobacco. Nitrate reductase is an important component; it is a
substrate-inducible enzyme and its activity is affected by the nitrate-
nitrogen concentration of leaves and, therefore, the amount of
nitrogen available in the soil (Long & Weybrew, 1981). The enzyme
has been used to assess nitrogen metabolism of plants and there is a
very good negative relationship between its activity and the
accumulation of starch in the leaves (Weybrew & Woltz, 1975). In
flue-cured tobacco that has been correctly fertilized and has received
adequate rainfall, there is a rapid depletion of soil nitrate and nitrate
reductase activity in leaves at about flowering. Starch then
accumulates rapidly as the demand for photosynthate in the reduction
process decreases, and the tobacco achieves a good balance between
the nitrogenous and carbohydrate components (a sugar:nicotine ratio
of about 6 to 8:1). Underfertilization and when rain has leached
nitrogen through the soil profile cause the transition from nitrate to
carbohydrate metabolism to occur before flowering, which results in a
larger than normal accumulation of starch and smaller total nitrogen
and nicotine concentration (a sugar:nicotine ratio of >9:1). However,
in drought and over-fertilized conditions, the relationship is reversed
because nitrogen is available for an extended period and the nitrate
reduction continues for a longer time (sugar:nicotine ratio of <5:1)
(Fig. 4.3) (Long & Woltz, 1972; Weybrew & Woltz, 1975; Weybrew,
1979; Long & Weybrew, 1981; Weybrew, et al., 1983). Flower (1996)
reported that the nitrate-nitrogen concentration of the leaf and the
nitrate reductase activity decreased at about the same time whether or
not nitrogen fertilizer was still being applied and that the high
nitrogen treatments had a greater enzyme activity. Factors that could
have been involved include a decrease in the rate of expansion of the
leaf and, therefore, in the demand for nitrate-nitrogen and an increase
in root growth and nicotine synthesis as a result of topping. Attempts
have been made to exploit the relationship between nitrate-nitrogen
and starch metabolism in an effort to produce tobacco with
prespecified chemistry levels, but limited success has been achieved
(Ismail & Long, 1980).
Effect of Nitrogen on Growth, Yield, Quality and Chemical
Composition
Tobacco is very sensitive to nitrogen nutrition. The crop must have
adequate nitrogen available during the growing phase to ensure
vigorous growth, and soil
 Fig. 4.3
Effect of leaching/underfertilization and drought/over-fertilization on
nitrate reductase activity and starch accumulation in tobacco (derived
from Weybrew, 1979).

 Page 81
nitrogen should be depleted soon after topping so that leaves ripen
correctly. With adequate moisture, an increase in the supply of
nitrogen results in an increase in leaf area and a decrease in
weight/unit area or thickness which is expressed as a higher filling
value in the cured leaf (Raper & McCants, 1967; Elliot & Court,
1978). Thus, a liberal supply of nitrogen is desirable when the object
is to produce a large thin leaf, such as required for cigar wrapper,
whereas a moderate supply is necessary for the development of the
flue-cured type. which has thicker leaves.
Excessive soil nitrogen will generally produce cured leaves which are
dark brown to black in color, dry and chaffy, and have a strong and
pungent smoke. In the field, a deficiency of nitrogen causes premature
yellowing of leaves, which when cured are generally pale in color,
close grained and thick bodied, and their smoke is flat and insipid. An
increase in nitrogen will increase the yield of tobacco, but quality is
often reduced at high levels (Elliot, 1970b; Court, et al., 1984).
Nitrogen is a component of the nicotine molecule and is important in
the synthesis of this constituent of tobacco. The concentration of
nitrogen in leaves is positively correlated with nicotine and negatively
correlated with starch and sugar concentrations (Sheen, et al., 1973;
Elliot & Court, 1978). Increasing the rate of nitrogen fertilizer
increases the amount of ash and concentrations of nicotine, total and
protein nitrogen, resin and petroleum ether extract, and decreases the
sugar concentration (McCants & Woltz, 1967; Elliot & Court, 1978;
Ryding, 1981). Nitrate-nitrogen concentration in the cured leaf is
affected by the rate of nitrogen fertilizer, soil moisture, soil type and,
to a lesser extent, by genetic differences (Broaddus, et al., 1965;
MacKown, et al., 1984). Miner and Sims (1983) reported that nitrate-
nitrogen began to accumulate in the leaves of burley tobacco when the
total nitrogen concentration reached 2.2% on a dry matter basis.
Nitrogen deficiency has been shown to increase the scopolin and
chlorogenic acid concentrations, and the total polyphenol
concentration decreased with increasing nitrogen fertilization
(Armstrong, et al., 1970). Court and Elliot (1978) found that in flue-
cured tobacco scopoletin was positively related and chlorogenic acid,
neo-chlorogenic acid, 4-caffeoylquinic acid, rutin and scopolin
concentrations were inversely related to the rate of nitrogen
fertilization. Another study with air-cured tobacco types also showed
an inverse relationship between the accumulation of phenolic
constituents and nitrogen fertilization (Sheen, et al., 1973). On the
other hand, Tso, et al. (1967) found that the amount of phenolic
compounds, including chlorogenic acid, rutin, scopoletin and
scopolin, were positively correlated with the rate of nitrogen fertilizer.
The differences reported in the various studies may be a reflection of
differences in tobacco type, curing method, environmental conditions
and chemical methods used to determine the phenolic constituents. In
a study by Court, et al. (1984), the application of nitrogen fertilizer
was found to increase hexane extracts, cytoplasmic lipids,
carotenoids, chlorophyll, neophytadiene and solanesol and to decrease
duvatrienediols; it had no effect on surface waxes, hydrocarbons and
total phytosterols.
Tobacco alkaloids and nitrite (nitrate derived) are the major precursors
of the tobacco specific N-nitrosamines (TSNA): N-nitrosonornicotine
(NNN), N-nitrosoanatabine (NAT), N-nitrosoanabasine (NAB) and 4-
(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). These N-
nitrosamines, which are present in both tobacco leaf and smoke, are
known carcinogens (MacKown, et al., 1984). MacKown, et al. (1984)
altered the lamina nitrate-nitrogen concentration of leaf by imposing
selected nitrogen fertility regimes having different application dates
and rates. Concentrations of TSNA were positively correlated (p £
0.01) with the concentrations of nicotine and nornicotine but not with
nitrate-nitrogen, despite a greater than 10-fold difference in nitrate
concentrations among treatments. However, these authors noted that
nitrate-nitrogen in tobacco products should not be ignored, as there is
ample evidence, such as that of Sims, et al. (1979a), to indicate that
nitrate-nitrogen levels are strongly associated with the formation of
oxides of nitrogen and the concentration of TSNA in smoke. In other
work, an increase in the rate of nitrogen fertilization increased the
concentration of NNN in a normal flue-cured tobacco (NC 2326), but
decreased it in a converter tobacco (NIC 117H-1) (Chamberlain, et al.,
1984).
Effect of Timing of Nitrogen Application
Raper and McCants (1967) reported that the presence of adequate
nitrogen in the leaves during the early stages of development, which
included cell division and early cell elongation, was of critical
importance in determining the final area of the leaf. Consequently, the
timing of nitrogen availability in the soil was critical, and information
on the alkaloid, sugar and nitrogen concentrations of selected leaves
indicated that a lack of nitrogen during a particular stage of growth
caused an increase in the reducing sugar concentration and a decrease
in the total alkaloid concentration in the leaves

 Page 82
produced during that stage. A relief of the stress restored the sugar and
total alkaloid balance in leaves produced subsequently. McCants and
Woltz (1967) concluded that tobacco requires a high percentage of the
total available nitrogen during the early stages of plant growth and the
amount should rapidly diminish during the later phase at about
topping. However, it has been reported that in under-fertilized
situations and for certain flue-cured tobacco cultivars, a relatively
small amount of nitrogen (< 15 kg N/ha), applied at or soon after
topping, improves the color and balance of the nitrogenous and
carbohydrate constituents in the cured leaf, without greatly affecting
the rate of ripening (Tobacco Research Board, 1993; Tobacco
Research Board, 1994a).
Effect of Nitrogen Source
Much work has been done on the effect of nitrogen sources on the
yield and quality of tobacco and often with conflicting results.
Initially, McCants and Woltz (1967) stated that tobacco grown in sand
and solution culture with nitrate was superior to that grown with the
ammonium form of nitrogen. In an analysis of fertility experiments
with flue-cured tobacco, these authors found that in 13 out of 15
experiments the value per acre of tobacco was greater where the
fertilizer contained at least 50% of the nitrogen in the nitrate form;
this limit on the proportion of ammonium has been recommended by
Collins and Hawks (1993). Rhoads (1972) found that a similar
proportion of nitrate was required for best yield and quality of cigar
wrapper tobacco. However, in Canada it is advised that only 25% of
the total nitrogen need be in the nitrate form (Ontario Ministry of
Agriculture and Food, 1994). McCants and Woltz (1967) concluded
that the relative response of tobacco to application of the ammonium
and nitrate forms depended on the extent and speed with which the
ammonium was converted to nitrate, which is dependent upon the
environmental conditions for biological nitrification. It has also been
found that plants supplied with ammonium absorb fewer cations, such
as calcium, magnesium and potassium, than those supplied with
nitrate (Collins & Hawks, 1993).
Other studies have shown few differences between the sources of
nitrogen. Tisdale, et al. (1952) tested various nitrogen sources,
including sodium nitrate, ammonium nitrate and urea, and found that
they had no effect on the yield or value of tobacco. In Zimbabwe
(Tobacco Research Board, 1986e) no differences in the yield, grade
index and concentration of reducing sugars, total alkaloids and total
nitrogen were found between sodium nitrate, ammonium nitrate and
urea. In a review, Chaplin and Miner (1980) reported that no
difference was found in the concentrations of reducing sugars, total
alkaloids and inorganic cations in cured leaves between urea and
sodium nitrate. Increasing the proportion of total nitrogen as urea
increased the chloride concentration in the green tissue, possibly as a
result of anion accompaniment of the absorbed ammonium-nitrogen.
Urea is often regarded as similar to an ammonium source of nitrogen,
but it does differ in being easily leached when first applied to the soil
because it is very soluble and is just as mobile as nitrate. However,
urea is converted quickly to ammonium compounds through
hydrolysis by the enzyme urease, after which the ammonium ions are
transformed to the nitrate ions by normal biological activity in the
soil. Thus, serious losses of nitrogen from the soil are avoided (Cooke,
1972). In another study on flue-cured tobacco, Elliot (1970b) found
that increasing the proportion of ammonium to nitrate from 0% to
100% significantly increased the yield, grade index, reducing sugars
and decreased the total nitrogen, total alkaloids and sand free ash, and
had no effect on the potassium content of cured leaves. The lower
concentrations of the nitrogenous constituents were attributed to a
dilution effect from greater yields. The positive response to
ammonium was associated with greater rainfall and was probably due
to the ammonium form being less susceptible to leaching than the
nitrate form. The work of Ryding (1968) and Tillet (1966) confirms
that ammonium-nitrogen is generally more efficient than nitrate-
nitrogen in a wet season.
b
Phosphorus
In contrast to nitrogen and potassium, absorption of phosphorus
occurs at a fairly constant rate throughout the growing season (Fig.
4.1). The difference in the absorption curve is due primarily to
differences in the relative availability of the elements in the soil, as
phosphorous is less soluble than the other two nutrients and is
released constantly into the soil solution throughout the growing
season as it is absorbed by the plant. Although many of the sandy soils
commonly used for tobacco production are inherently low in
phosphorus, deficiency symptoms are rarely observed due to a build
up of the element from repeated heavy applications. The early growth
response to applied phosphorus, so frequently observed, may result
from the influence of temperature on absorption. Parups, et al. (1960)
found that at a soil temperature of 14°C there was a greater response
to added phosphorus than at

 Page 83
21°C and 30°C, when the increase in growth was very small.
Nonetheless, the average plant weight was considerably lower from
plants grown at 14°C. At the lower temperature, the phosphate
concentration in the leaf indicated deficiency levels. Parups and
Nielsen (1960) concluded that phosphorus was the most important
element for growth of tobacco at low temperatures.
The absorption of phosphorus by tobacco roots is also influenced by
soil pH. In the absence of calcium, the uptake of phosphorus was
found to increase from pH 4 to pH 6, but then decreased with a
minimum value at pH 8. With calcium present, the uptake of
phosphorus increased up to pH 5, but at higher pH values the
precipitation of calcium phosphates occurred, thus limiting the
availability of phosphorus to the plant (McEvoy, 1964).
It is well established that rapid and vigorous development of young
plants is stimulated by a high level of available phosphorus (McCants
& Woltz, 1967). Growth responses to applied phosphorus are more
often observed early in the growing season than they are in final yield
and quality. However, Ryding (1986) reported a pronounced response
in yield and quality to applied phosphate on phosphorus deficient
soils, and that the response was increased when an adequate amount
of potash was also applied. Lolas, et al. (1978) found a similar yield
response to applied phosphate on soils low in available phosphate, but
the quality was not affected.
A deficiency of phosphorus results in stunted growth, poor leaf
expansion, unusually dark-green leaves and occasionally there may be
numerous white spots on the lower leaves. The cured leaves tend to be
dark brown or, because they do not mature normally, may be dark
greenish in color and lack the lustre of normal leaves (McCants &
Woltz, 1967). Low soil phosphorus levels also result in a decrease in
the phosphorus, nitrogen and magnesium concentrations in the leaf
and delayed maturity (McEvoy, 1951). An excess of phosphorus in the
soil can contribute to deficiencies of copper, iron and zinc (Gilbert,
1952; Cooke, 1967; Elliot, 1975a).
No consistent relationship has been established between rates of
phosphorus and the organic constituents of the cured leaf (Collins &
Hawks, 1993). Crafts-Brandner, et al. (1990) found that starch
accumulation in green tobacco leaves of the burley cultivar KY 14
increased with decreased levels of phosphorus, whereas with the flue-
cured cultivar Speight G28 and N. rustica cultivar Pumila, starch
increased and then remained relatively constant with increased
phosphorus. In flue-cured tobacco, applied phosphorus had little effect
on the equilibrium moisture and filling value or the concentration of
nicotine, sugars and resin of the cured leaf (Ryding, 1981). Other
studies have found a positive relationship between phosphorus and
sugar concentration (Tso, 1990c). There are contradictory results on
the effect of phosphorus on the total alkaloids (McCants & Woltz,
1967). Therefore, the application of phosphorus does not have a
consistent effect on the concentrations of sugar and nicotine in the
cured leaf and the discrepancies reported in the literature probably
reflect the conditions under which the experiments were done.
c
Potassium
The importance of potassium in the nutrition of tobacco is illustrated
by its application in all production areas, and the fact that the uptake
of this nutrient is the greatest of all mineral elements (Raper &
McCants, 1966). Tobacco is known to be a luxury user of potassium
and the amount of potassium fertilizer used often exceeds normal
yield requirements (Tso, 1990c). It was found by McEvoy (1955) that
when potassium was deficient, sodium partially replaced potassium in
the nutrition of flue-cured tobacco plants. Furthermore, that the two
elements had an additive effect in depressing the uptake of calcium,
magnesium and phosphorus. When the nutrient supply of potassium is
low, sodium will delay the appearance and lessen the severity of the
potassium deficiency symptoms (Hutcheson, et al., 1959). However,
the response to sodium was not considered adequate to suggest that
any of the potassium or potash (K2O) in tobacco fertilizers was
replaced by sodium.
Potassium deficiency symptoms occur at concentrations of less than
1% of dry matter and tend to be aggravated by dry weather
(McMurtrey, 1964; Green 1971). In the initial stage, potassium
deficiency is manifested by a slight mottling and the appearance of
brownish yellow specks, especially near the tip and margins of the
leaves. When the deficiency is acute, the spots become brown and
result in large dead areas. The dead tissue may fall out and give the
leaf a ragged appearance. When deficiency occurs in small plants, the
symptoms appear first on the lower leaves and spread progressively to
the upper leaves. However, if deficiency occurs near topping, the
symptoms may appear first on the upper leaves. Potassium deficiency
may be accentuated by excess nitrogen, particularly in the ammonium
form, and by high levels of magnesium or sulphur (McCants & Woltz,
1967). A liberal supply

 Page 84
of potassium is needed from transplanting to 2 weeks before
harvesting begins, and this will be sufficient to meet the requirements
for the remaining development of the plant without any further
absorption. However, if the supply is stopped 4 weeks prior to
harvesting, the reserve in the plant is inadequate and deficiency
symptoms occur (McCants & Woltz, 1967). With moderate
fertilization, potassium is distributed uniformly in all leaf positions,
whereas high rates promote accumulation in the lower leaves
(Gribbins, et al., 1941).
Leaf color, texture, combustibility and hygroscopic properties are
believed to be enhanced by potash fertilizer (Tso, 1990c). Tibbitts
(1962) reported that the ignition temperature of cigar tobacco was
decreased significantly as the potassium concentration in the leaf
increased. Bowling and Brown (1947) found that in Maryland tobacco
successive increments of potash fertilizer resulted in larger and
slightly narrower leaves with a decreased weight per unit area when
cured. The yield, quality, fire-holding capacity, hygroscopic properties
and concentration of total ash and potassium were also improved.
With flue-cured tobacco, the opinion is widely held that a high level
of potassium results in a deep orange color; however, numerous
experiments have failed to prove this view (McCants & Woltz, 1967).
Also, it is believed that increased applications of potassium produce
tobacco which is thinner and more elastic and pliable; however, in
controlled experiments using different rates of potassium these
physical differences are often difficult to measure (Collins & Hawks,
1993). Ryding (1981) found that potash applications up to 50 kg/ha
improved yield, financial return and sometimes quality, and higher
rates had little further benefit. Chaplin and Miner (1980) found that
applying more potassium to soils with >0.33 milliequivalents (meq)
potassium/100 ml had no effect on the yield or chemical properties of
flue-cured tobacco, whereas on soils that contained 0.12 meq
potassium/100 ml, visual deficiency symptoms were observed at the
potash rates of 135 kg/ha or less. Increased rates of potassium had no
significant effect on yield, grade index, total alkaloids, reducing
sugars or ash. Burn rate increased and nicotine delivery decreased
with increased potassium, but differences in filling value, equilibrium
moisture, tar delivery and resistance to draw were not significant.
Elliot (1968) found that for flue-cured tobacco, rates or time of
application of potassium had no effect on the 3-year averages of the
concentration of magnesium, total ash, total nitrogen, total alkaloids,
reducing sugars and petroleum ether extracts, and the indices of
shatter resistance, filling value, grade and maturity. In burley tobacco,
Link and Terrill (1982) found that the concentration of potassium in
the cured leaf was increased but there was no yield or quality response
to applied potassium on soils high in potassium.
d
Calcium
Calcium is one of the principal inorganic constituents of tobacco and
next to potassium is absorbed in the second largest quantity (McCants
& Woltz, 1967). The role of calcium in plants is not clear, but it is
required for functions such as cell division and expansion and
chromosome stability (Elliot, 1975a). Calcium does not move from
the older to younger parts of the tobacco plant and, therefore, calcium
deficiency begins at the growing point. Leaves of calcium-deficient
plants are dark green and there is a hooking downward at the tips and
margins, and in the advanced stage the terminal bud dies (McCants &
Woltz, 1967). If the calcium deficiency does not become acute until
the flowering stage, there is a tendency for the plant to shed blossoms
and buds, and the flowers that remain show dieback of their corollas
with the pistils protruding (McMurtrey, 1964). The uptake of calcium
is reduced by applied potassium (Hutcheson, et al., 1959), and the
omission of potassium was found to increase the concentration of
calcium in leaves (Elliot, 1975a). It has been shown that the calcium
concentration in leaves is directly correlated with leaf position and
weather fleck, and the lowest leaves had the highest concentration of
calcium and the greatest amount of weather fleck (Trevathan &
Moore, 1981). However, rate of calcium fertilization (CaSO4) was not
related to the calcium concentration in leaves and it was concluded
that it would do little to decrease the amount of weather fleck.
Lime is the principal source of calcium and, except for Darkis, et al.
(1937), there is general agreement that dolomitic lime, which contains
both calcium and magnesium, will improve the quality of tobacco.
Peedin and McCants (1977) found that lime (dolomitic limestone) was
more effective than fertilizer calcium (CaSO4) in increasing the
calcium level of cured leaves. Yield and value/ha were increased by
lime application to soil with 40% aluminum saturation and pH 5.3. A
significant response to fertilizer calcium was obtained only on a soil
with 0.4 meq exchangeable calcium/100 ml soil. The average price
was not affected by lime application, whereas the 168 kg/ha rate of
fertilizer calcium decreased average price; the inferior leaf quality, as
measured by average price, was not associated with excessively high
leaf concentrations of

 Page 85
calcium and magnesium. Cured leaves from limed plots contained
higher concentrations of calcium, magnesium and nitrogenous
constituents and generally lower concentrations of potassium and
reducing sugars. These authors suggested that aluminum was
neutralized by the lime and that this resulted in larger, more efficient
root systems that could recover a greater proportion of applied
nitrogen. Ryding (1967) found that lime increased both yield and
quality of flue-cured tobacco, and the effect was more marked at a
low soil pH of about 4.2 (0.01 M CaCl2).
e
Magnesium
Magnesium deficiency, commonly called 'sand drown', results in an
interveinal chlorosis of tobacco leaves. It normally occurs on sandy
soils low in exchangeable magnesium and a few acid soils low in
exchangeable magnesium, as well as being induced by high levels of
potassium (Wild, 1988). Mulchi, et al. (1967) found that in flue-cured
tobacco, magnesium deficiency was common in seasons of heavy
rainfall, and the symptoms were seen when the concentration of
magnesium in the leaf was £ 0.2% of dry matter. Plants that show
magnesium deficiency symptoms have a lower percentage of
magnesium in the bottom than in the top leaves, whereas in normal
plants the opposite occurs (McMurtrey, 1947). As the deficiency
symptoms become worse, the starch and sugar concentration declines
and there is an increased concentration of organic acids in the leaves.
Also, there is an increase in the ash content, probably as a result of the
decrease in carbohydrates (McMurtrey, 1947). Magnesiumdeficient
cured leaves are usually dark with irregular colors and are abnormally
thin and nonelastic (McMurtrey, 1964). A certain percentage of
magnesium in the leaf appears to be essential for satisfactory
combustion in cigars and the optimum concentration is nearly 2%;
greater concentrations decrease the combustibility (Anderson, et al.,
1931). Magnesium deficiency is easily corrected with dolomitic lime
(Mulchi, et al., 1967).
f
Sulfur
Sulfur-deficient tobacco plants show a mild chlorosis over the whole
plant which is similar to nitrogen deficiency (Elliot, 1975a). A
distinguishing difference is that sulfur-deficient plants do not lose
their leaves by 'firing', they tend to pale from top to bottom and they
do not respond to additional nitrogen (McCants & Woltz, 1967;
Collins & Hawks, 1993). Sulfur deficiencies are becoming more
common because sulfate is subject to leaching and some fertilizers
contain little or no sulfur (Smith, et al., 1987; Collins & Hawks,
1993). Smith, et al. (1987) obtained yield responses to applied sulfur
at two of four locations with argillic horizons 0.45 m deep and soil
sulfate-S concentrations of 2 to 5 g/m3 in the upper 0.45 m. Sulfur had
no effect on the quality of tobacco in these experiments, however, in
other studies, excess sulfur was found to adversely affect quality by
producing grey tobacco (Ryding, 1968). Sims, et al. (1979b) found
that high levels of sulfate in the rooting medium decreased nitrate
reductase activity and molybdenum accumulation in burley tobacco.
They concluded that additions of sulfate may have a detrimental effect
at low levels of molybdenum, but not at high levels. Neas (1953)
showed that the concentrations of reducing sugars and total alkaloids
were independent of the rate of applied sulfur. Although a high sulfur
concentration in tobacco has been inferred to have detrimental effects
on leaf burn, the limited amount of data available is contradictory
(McCants & Woltz, 1967)
g
Other Nutrients
Boron is essential for tobacco growth and a deficiency causes the
young leaves of the terminal bud to become a bright green color, but
paler at the base than the tip. The tissues at the base of the young
leaves show signs of breakdown and distortion of the leaves occurs,
followed by death of the terminal bud. Boron may participate either
directly or indirectly in the synthesis of lignin (McCants & Woltz,
1967), and it appears that boron is required by those plants in which
there is a well developed and lignified xylem (Lewis, 1980). It has
been observed that boron deficiency produces plants with weak cell
walls (Loomis & Durst, 1992). Zvomuya (1994) showed that both soil
and foliar applications of boron increased the thickness of lignified
tissue in the midrib and it reduced the amount of leaf breakage in
tobacco. Boron-deficient plants have a higher concentration of starch
and sugars than healthy ones, presumably due to the obstruction of
transport through the disorganized phloem (van Schreven, 1934).
Boron is required by tobacco in very small quantities and excesses are
phytotoxic (Collins & Hawks, 1993). In Zimbabwe, it was reported
that 9 kg borax/ha (1.02 kg boron/ha) depressed yield and quality, and
larger amounts severely damaged plants (Tobacco Research Board,
1966).
Molybdenum is an essential constituent of the nitrate reductase system
in plants. A very small amount

 Page 86
is required by tobacco plants in comparison with most of the other
micronutrients (Elliot, 1975a). The major factor influencing the
availability of molybdenum is soil acidity. The availability of this
nutrient increases as soil pH increases (Miner & Sims, 1983). Sims
and Atkinson (1976) reported an interaction between lime and
molybdenum rates, where highest yield and molybdenum
concentrations of cured leaves occurred in the presence of both
additional lime and molybdenum. An interaction occurs between
sulfur and molybdenum with applications of sulfur decreasing the
uptake of molybdenum (Sims, et al., 1979b). Although deficiencies of
molybdenum have not been found under field conditions (Collins &
Hawks, 1993), Chaplin and Miner (1980) suggest that molybdenum
availability may be limiting in some tobacco soils and, since its status
cannot be predicted by soil tests, it may be a form of 'hidden hunger'
causing reduced yield and quality in some instances. It has also been
suggested that on certain burley soils of low pH, improvements in
yield and quality from molybdenum fertilization may be due partly to
a reduction in manganese toxicity (Miner & Sims, 1983).
Chloride is generally recognized as an essential micronutrient (Broyer,
et al., 1954), and there is considerable evidence that beneficial effects
arise with tobacco from the presence of small amounts of chloride in
the fertilizer (McCants & Woltz, 1967). Neas (1959) found that small
amounts of fertilizer chloride improved the yield and quality of flue-
cured tobacco. This work also showed an increase in the moisture
content of the leaf with increase in chloride content. The highest
concentration of chloride is in the lower leaves and it decreases
progressively to the top of the plant. It is generally thought that
concentrations in excess of 1% can produce poor quality tobacco.
The uptake of chloride by plants increases linearly over a wide range
of concentrations in the substrate (Reisenauer & Colwell, 1950), and
an excess produces a leaf which is greatly thickened and exceedingly
brittle, the leaf margins curl upwards and the leaf presents a
distinctive sleek, glabrous appearance. In the cured leaf, excess
chloride produces muddy, dingy and uneven colors and an undesirable
odor known as 'wet dog' (McCants & Woltz, 1967). Neas (1961)
found an increase in the proportion of poor leaf grades and
equilibrium moisture, and a decrease in the alkalinity of the water-
soluble ash and duration of burn with increasing rates of fertilizer
chloride. It is thought that the effect of chloride on burn is through an
increase in the hygroscopic substances in the leaf (McCants & Woltz,
1967). The deleterious effects of excess chloride can be reduced by
the addition of lime (McCants & Woltz, 1967) and the use of nitrate,
rather than ammonium sources of nitrogen (Skogley & McCants,
1963).
High concentrations of chloride in the media decreased the
accumulation of nitrate-nitrogen, phosphorus and sulfur in the leaves
of greenhouse grown tobacco (Skogley & McCants, 1963). It was also
observed that protein nitrogen was decreased by chloride when either
ammonium or nitrate nitrogen was used. A positive correlation
between chloride and starch and sugars has been reported, and it was
concluded that the effect of chloride on carbohydrate metabolism was
because of its effect on the amylytic enzyme in the leaf, which
resulted in the marked accumulation of starch when the chloride
content was high (Garner, et al., 1930; Peele, et al., 1960).
Topping and Suckering
Topping is the removal of the inflorescence and uppermost leaves in
order to stimulate the growth and development of the remaining top
leaves. This practice breaks apical dominance and the axillary buds on
the stalk begin to grow. The removal of the buds by hand or
chemically is known as suckering, and it ensures that the top leaves
obtain sufficient nutrients for their development.
Topping increases the area, weight/unit area and nicotine
concentration and decreases the filling value of the upper leaves
(Elliot, 1975b; Akehurst, 1981a). Consequently, the height and time of
topping and their effects vary considerably according to the type of
tobacco. Also, it must be remembered that the final cured leaf product
is an interaction of numerous factors which include time and height of
topping, climate, plant population, fertility and method of curing.
Dark air-cured and fire-cured tobaccos are topped early and very low
at 8 to 15 leaves in order to prolong the vegetative period and the
accumulation of nutrients and nicotine. The resultant cured leaf tends
to be large, heavy, oily, tough, leathery and dark in color. Intermediate
heights of topping are used to obtain less drastic effects. Flue-cured
tobacco is topped to about 18 leaves and the leaf has a moderate
thickness, area and nicotine concentration. Burley tobacco is topped to
about 24 leaves and grown on fertile soil which produces a high yield
of well developed, light-bodied leaves with a medium-to-high nicotine
concentration. In the case of shade-grown cigar wrapper leaf, topping
is delayed and only the flower bud is removed. Oriental

 Page 87
tobacco is not topped because the leaves should be small and have a
low nicotine concentration.
a
Effect of Topping on the Growth, Physiology and Chemical
Composition of Leaves
Topping has a much greater effect on the chemical and physical
properties of younger than older leaves. For example, the increase in
leaf area is confined to leaves that are less than 85% expanded at the
time of topping and the younger the leaf the greater the effect on its
ultimate area (Wolf & Gross, 1937; Papenfus & Quin, 1984). Topping
has little or no effect on the leaves that are mature, such as those on
the lower half of the stalk.
The upper leaves of topped plants are much larger, thicker and
heavier-bodied than those from untopped plants (Avery, 1934;
Papenfus, 1970). Avery (1934) selected leaf numbers 17, 19 and 20/21
as representative of the upper leaves of topped and untopped plants.
Leaf 17 showed little response to topping, leaf 19 showed a definite
response and leaf 20/21 a marked response. The leaves from topped
plants were about 32% greater in area than corresponding leaves from
untopped plants. The average thicknesses of leaves 17, 19 and 20/21
from topped plants were 184, 251 and 308 µm, respectively, and the
corresponding leaves from untopped plants were 188, 200 and 188 µm
thick, respectively. The increased growth of the upper leaves was due
to a greater than usual increase in cell size, the palisade and the upper
and lower epidermal cells averaging 31% larger than in untopped
plants. Similar results were obtained by Wolf and Gross (1937) who
showed that topping increased the size of the epidermal and palisade
and spongy parenchyma. Avery (1934) reported that the only change
in the number of cells was found in the vascular tissue, particularly
the xylem, which had an average of 47% more lignified xylem
elements in the petiolar bundle. It was noteworthy that there were
20% fewer sieve tubes and companion cells in the external phloem in
the petiolar bundles of the twenty-first leaves of topped plants than in
the corresponding leaves of untopped plants, a condition quite
opposite to the numbers of lignified xylem elements. Avery contended
that the absence of the developing seeds and the subsequent cessation
of diversion of material from leaves to seeds was accompanied by a
cessation of the differentiation of the phloem cells.
Topping has many other effects, some of which are not understood. It
improves drought tolerance and slows leaf senescence so that leaf area
duration is extended (Papenfus, 1970; Papenfus & Quin, 1984).
Papenfus (1967) found that topping increased the rate of net
photosynthesis/unit area of the upper leaves. Crafts-Brandner (1991)
showed that the loss of chlorophyll and ribulose-l,5-bisphosphate
carboxylase-oxygenase (Rubisco) activity, which indicates aging, was
slower in topped plants compared with those untopped. In contrast to
chlorophyll and Rubisco activity, the activities of the key enzymes
involved in starch and sucrose synthesis remained relatively constant
over time for untopped plants but were enhanced in topped plants.
Also, Crafts-Brandner (1991) concluded that the upper leaves of
topped plants functioned as both source leaves with the enhanced
ability to synthesize carbohydrate, and sink leaves with enhanced
growth.
Leaves from topped plants have a higher concentration of starch than
untopped plants, which results in an increased reducing sugar
concentration in the cured leaves (Chaplin, 1967; Crafts-Brandner,
1991). Marshall and Seltmann (1964) found that earlier topping
produced a higher concentration of reducing sugars in the leaves than
later topping, whereas Elliot (1966) reported that time of topping had
no effect on reducing sugars. In these experiments by Marshall and
Seltmann and also Elliot, the earliest topping was done at about the
extended bud stage when the first floral buds were well formed. Elliot
(1975b) also found that when topping was done very early at the pre-
bud stage, the concentration of starch and reducing sugars was lower
than when topped at about the extended bud stage; topping even later
resulted in a decrease in the concentration of reducing sugars. These
results were confirmed in research in Zimbabwe (Tobacco Research
Board, 1988b; Tobacco Research Board, 1992). Further studies in
Zimbabwe on the effect of topping on the nitrogen and carbohydrate
metabolism of fluecured tobacco have been done (Tobacco Research
Board, 1994a; Tobacco Research Board, 1995a). This work showed
that topping increased the initial rate of starch accumulation, both in
early-and late-topped plants compared with the untopped treatment
(Fig. 4.4), and had no effect on nitrate reductase activity. Rapid
accumulation of starch in the upper leaves of all topped treatments
began at about the same time as the rate of leaf expansion declined,
and this was also observed by Crafts-Brandner (1991). Initially, the
increase in starch concentration was largest in the early-topped
treatment; however, the build-up in the late-topped treatment reached
a similar value soon after it was topped. At this time, the lowest
concentration occurred in the untopped treatment. As ripening
progressed from about 115 days after planting, there

 Page 88

Fig. 4.4
Effect of topping on the starch concentration of the upper leaf in flue-cured
tobacco (Tobacco Research Board, 1995a).
was a drop in starch accumulation which was greatest in the early-
topped treatment (Fig. 4.4). As a result, the late topped plants had the
highest starch concentration. In all treatments, the concentration of
starch in the green leaves was much higher than that of reducing
sugars, and the sugars increased with ripeness and were highest in the
untopped plants.
The increase in starch accumulation after topping may have been due
to the removal of a major nutrient sink. It is also possible that topping
stimulated the synthesis of starch by delaying senescence and
increasing the activity of the enzymes involved in starch and sucrose
synthesis (Crafts-Brandner, 1991). The drop in starch accumulation
during the latter part of ripening and the increase in reducing sugars
are characteristics of tobacco senescence (Long & Weybrew, 1981).
However, the more distinct decline in starch concentration in the
early-topped treatment is more difficult to explain. A number of
factors might be involved including various senescence processes and
the translocation of carbohydrate out of the leaf, possibly to maintain
a more vigorous root system and nicotine synthesis.
It is generally accepted that topping increases root growth. Papenfus
(1970) found that early topping increased the dry mass of roots by
about 42% and late topping by 12% of the untopped control. Steinberg
and Jeffrey (1957) concluded that topping and suckering increased the
size and branching of the root system. Further evidence for the effect
of topping on root growth is the significant increase in nicotine which
occurs after topping, as nicotine is synthesized in the roots and
translocated to the leaves (McCants & Woltz, 1967; Bush, 1981).
However, some data indicate that topping has no effect on root growth
(Osmond & Raper, 1982; Crafts-Brandner, 1991). The lack of
response may have been due to topping relatively late.
Topping significantly increases the concentration of nicotine in leaves
and the earlier that plants are topped the greater the increase (Marshall
& Seltmann, 1964; Elliot, 1966; Chaplin, 1967). The nicotine
concentration of the cured leaf is usually increased by more than that
of the reducing sugars and as a result of this the ratio of sugar:nicotine
is decreased by about 30% (Papenfus & Quin, 1984).
Elliot (1966) and Marshall and Seltmann (1964) found that time of
topping had no effect on total nitrogen concentration. Other results
support this, except when the tobacco is topped at an early stage when
noticeable increases in total nitrogen occur (Elliot, 1975b; Tobacco
Research Board, 1992; Tobacco Research Board, 1994a; Tobacco
Research Board, 1995a). The effects of topping on total nitrogen
concentration are not as dramatic as on nicotine, therefore the ratio of
total nitrogen:nicotine decreases, and the earlier the plants are topped
the smaller the ratio.
Although topped plants generally have a lower concentration of
petroleum ether extracts than untopped ones, the time of topping has
little effect (Elliot, 1966; Elliot, 1975b). There is a greater
accumulation of duvatrienediols in topped compared with untopped
tobacco plants (Court, 1982). Topping increases the concentration of
most of the neutral volatile compounds associated with the overall
'character' of the smoke, particularly when suckers are controlled. The
practice, therefore, increases the proportion of flavor grades at the
expense of the filler grades (Weeks & Seltmann, 1986; Papenfus,
1987). Tobacco from topped plants has a higher tar (total particulate
matter) in the smoke stream, and the values are highest with early
topping. The effect on nicotine is greater than on tar, and
consequently, the tar:alkaloid ratio in smoke decreases with topping
(Elliot, 1975b; Papenfus, 1987; Tancogne, et al., 1996).
Decreasing the topping height progressively increases the response of
the remaining leaves, for example, the area, weight/unit area and
nicotine concentration of the upper remaining leaves are increased as
topping height is decreased (Elliot, 1970a; Campbell, et al., 1982).

 Page 89
b
Effect of Topping on Yield, Quality and Grade Distribution
The highest and lowest yields of cured tobacco are obtained from
early-topped and untopped plants, respectively. Topping also improves
the quality of tobacco, as measured by price or grade index, and
generally the best quality is obtained from early topping (Marshall &
Seltmann, 1964; Elliot, 1975b). Failure to top results in low yields of
inferior quality tobacco, particularly from the middle and upper
leaves. A low topping height will increase the dry mass of individual
leaves; however, this increase is usually insufficient to compensate for
the loss in leaves removed in the operation, and consequently, total
yield is decreased (Campbell, et al., 1982). Although yield is
decreased with lower topping, quality and price are normally
increased. Consequently, the highest value/ha is usually obtained with
an intermediate topping height (Elliot, 1970b; Tobacco Research
Board, 1984b).
Height of topping affects the proportion of lugs, cutters, leaf and tips.
If plants are topped low mainly lugs and leaf grades are obtained,
higher topping produces lugs, cutters, leaf and tips and no topping
results mainly in lugs and cutters.
Topping can, therefore, be used to modify the tobacco according to
the requirements of the market. For example, crops which are not
topped and are grown at closer than normal spacing produce small,
light-bodied, pale lemon-colored leaves with less than 1.5% nicotine
and with neutral aroma and taste (Papenfus, 1987).
c
Suckering
If suckers, axillary bud growth, are not controlled they_ have a severe
effect on the development and chemical composition of the upper
leaves, and many of the benefits of topping are offset by the growth of
suckers. Immediately after topping, sucker growth is very vigorous
and has the largest influence on yield, quality and chemical
composition (Cousins, 1988). Chemical agents (suckercides) control
suckers better than manual suckering, and a combination of these two
methods provides the best control (Tobacco Research Board, 1996).
Generally, the concentration of total alkaloids increases as the
efficiency of sucker control increases (Chaplin, 1967), except that
frequent manual suckering produces a higher concentration of nicotine
than suckercides, despite the fact that manual suckering produces a
greater dry mass of suckers (Cousins, 1988). Therefore, continuous
removal of the axillary buds probably stimulates root activity and
nicotine synthesis. Results have shown that the suckercide maleic
hydrazide (MH) increases the concentration of reducing sugars in
flue-cured tobacco compared with manual suckering (Chaplin, 1967).
Both MH and frequent manual suckering decrease the filling value,
total ash, alkalinity of the water-soluble ash and total ash, and increase
the equilibrium moisture content (Chaplin, 1967; Tso, 1990d).
Weeks and Seltmann (1986) found that good sucker control increased
the concentration of the neutral volatile constituents, and that there
was a positive association between sucker control and the flavour of
tobacco smoke. Goins, et al. (1993) reported that nonsuckered tobacco
was significantly lower in concentrations of duvatrienediols, and they
concluded that a high degree of sucker control, whether chemical or
manual, was important for high concentrations of duvatrienediols in
harvested tobacco.
Plant-Water Relations
While much research has been done on the effect of different
irrigation regimes on the growth of tobacco, there is a lack of
information on the physiological aspects of plant-water relations. It
should be realized at the onset that the effects of these relations per se
are difficult to determine, as there are so many other factors that
influence the growth of tobacco. This is illustrated by the differences
between rainfed and irrigated tobacco in Zimbabwe, where irrigated
crops are generally higher yielding and the cured leaves are thicker
bodied, with a higher ratio of sugar:nicotine. These differences are not
only due to the amount and distribution of the water, but also the
intensity of light, temperature, humidity and the availability of
nutrients (Stocks, 1994). This section discusses the effects of a deficit
or excess of water, from rainfall or irrigation, on the growth, yield,
quality and the plant-water requirements of tobacco. Other climatic
factors are reviewed in the section on responses to climatic factors.
a
Effect of Soil Water Deficit
Low soil moisture affects leaf width more than length and this results
in elongated leaves (Pearse, 1962). Clough and Milthorpe (1975)
found that the rate of leaf initiation in well watered tobacco plants was
constant with time, but was rapidly reduced by a small water

 Page 90
deficit and ceased at leaf-water potentials <-750 J/kg (-0.75 MPa).
Cell expansion showed a similar response, but cell division was much
less sensitive to water stress and continued at a decreased rate even
after leaf expansion had ceased. The fact that cell division continued
even after cell expansion had stopped explains why leaves can
subsequently attain areas similar to those that have been watered
constantly (Papenfus & Quin, 1984). Root growth and branching are
also decreased by dry conditions (Goodall, 1958).
It has been well documented that free proline accumulates in drought-
stressed plants because wilting inhibits proline oxidation. A positive
correlation also exists between absisic acid and proline, and an
application of absisic acid increases the concentration of free proline
in leaves. It seems that the increase of proline in dehydrated tissues
might be beneficial to the plant under stress (Lange, 1975). van
Rensburg and Kruger (1994) reported that proline concentrations in
droughttolerant cultivars increased sharply at a leaf water potential of
about -1.27 MPa and at -1.50 MPa in the droughtsensitive cultivars.
However, absisic acid accumulation preceded that of proline, and at a
leaf water potential value of -0.77 MPa, the absisic acid
concentrations of the droughttolerant cultivars were already
significantly greater than those of the droughtsensitive cultivars. The
value at which absisic acid and proline start accumulating rapidly and
the accumulated proline concentration were recommended as
selection parameters for drought tolerance in tobacco.
b
Effect of Water-Logging
Tobacco is very sensitive to water-logging and this is one of the
reasons for growing it on ridges. Plants grown under wet soil
conditions tend to become yellow, wilt and have decreased
photosynthesis (Goodall, 1958). Flooding essentially stops gas
exchange between the soil particles and the atmosphere, and seriously
impedes the flow of O2 to active root surfaces and to the micro-
organisms in the soil. The typical 'drowning' injury is not produced
solely by a lack of O2 or excessive amounts of CO2, but also by toxic
compounds, such as nitrite, derived under anaerobic conditions
(Harris & van Banvel, 1957). Using mixtures of O2 and N2, Hopkins,
et al. (1950) found that root growth was inhibited when the O2
concentration around the roots decreased to 0.5%, although top
growth and nutrient accumulation continued. An O2 concentration of
1.5% had no effect on root growth. However, Harris and van Banvel
(1957) showed that the presence of CO2 increased the effect of a
deficiency of O2 in the soil atmosphere. They demonstrated that the
functions of a plant were not appreciably reduced until the CO2
concentration became greater than the O2 concentration, and then they
were reduced considerably. Campbell (1973) found that the O2
concentration decreased from 16% to less than 4% in the soil
atmosphere within 24 hours after flooding, and after removal of
gravitational water increased to the original concentration within 48
hours. If flooding occurred for longer than 48 hours, the yield of
fluecured tobacco was significantly reduced to less than 40% of that
of the unflooded plants. Plants were more susceptible to flood damage
at the 12-leaf stage than at the 7-leaf stage. Reducing sugars increased
in tobacco leaves of plants flooded for either 12 or 24 hours, but
decreased significantly when flooded for either 72 or 120 hours. Total
alkaloids decreased at all times of flooding.
c
Effect of Plant-Water Relations on Yield, Quality and Chemical
Composition
It has been reported that irrigation improves yields, especially in dry
years, and more frequent applications of water but in smaller
quantities are desirable because leaching of nutrients is decreased
(Walker & Vickery, 1959; Tobacco Research Board, 1988c; Phillips,
et al., 1991; Collins & Hawks, 1993). On the other hand, over-
irrigation, particularly during early growth, significantly reduces yield
and quality of tobacco (Tobacco Research Board, 1978). Generally,
wet season or well watered tobacco has lower nitrate-nitrogen, total
nitrogen, total alkaloids and petroleum ether extract concentrations
and higher starch and reducing sugars than plants grown in a dry
season or under a slight moisture stress. The reasons for this are
leaching of soil nutrients, particularly nitrogen, and dilution due to
greater yields (Walker & Vickery, 1959; Ismail & Long, 1980;
Weybrew, et al., 1983; Papenfus & Quin, 1984; Tobacco Research
Board, 1987d; Phillips, et al., 1991; Reynolds & Rosa, 1995).
McCants and Woltz (1967) observed that tobacco did not produce the
desired leaf quality characteristics under moisture regimes that
resulted in rapid growth from transplanting to harvest. Also, plants
produced under moisture deficiency conditions which result in severe
reductions in growth did not develop the desired properties. They
deduced that a growth pattern between these two extremes would be
optimal. They hypothesized that a slight moisture stress at one or
more stages in the development of the plant would be advantageous
for the formation of a commercially

 Page 91
desirable leaf. More recent irrigation work does not substantiate this
hypothesis. Ismail and Long (1980) and Reynolds and Rosa (1995)
concluded that there must be sufficient water for unrestricted growth
for the best yield and quality. Research in Zimbabwe (Tobacco
Research Board, 1985b; Tobacco Research Board, 1987d) found that
yield and quality were decreased with sprinkler irrigation when plants
were stressed for extended periods or were heavily watered. The best
tobacco was produced when the plants received frequent, light
irrigations. These results suggested that the sprinkler irrigation regime
influences the growth and leaf attributes largely through the
differential leaching of soil nitrogen, rather than moisture stress per
se. Using drip irrigation, a treatment which applied water daily with
plants grown under no stress was compared with one where the soil
was depleted to 50% of available moisture before each irrigation. The
yield and quality of tobacco grown without stress were superior,
probably because the loss through leaching was compensated by
applying the nitrogen fertilizer through the drip irrigation system
(Tobacco Research Board, 1994b).
d
Water Requirements of Tobacco
A typical pattern of the water used by flue-cured tobacco is illustrated
in Fig. 4.5. The actual values depend upon the rate of growth, the
availability of water in the soil and the climatic conditions. Water use
by the plant increases to a maximum about 7 to 9 weeks after
transplanting and then decreases as the plants mature. An estimate of
crop water requirements (evapotranspiration, ET) can be obtained by
relating the reference crop evapotranspiration (ET0) to that of the crop
by using a crop coefficient (kc) (FAO, 1977).
 Fig. 4.5
Water use by tobacco (Harrison & Whitty, 1971).
Crop coefficients are linearly related to the canopy cover until the first
harvest, and as leaves mature they transpire less and the crop
coefficients decline faster than the crop canopy (Turner, 1969;
Papenfus & Quin 1984; Tonello, et al., 1993). Growth rates vary with
the type of tobacco, the environmental conditions and irrigation
regime or system used and, consequently, different crop coefficients
have been reported (Table 4.3).
Table 4.3 Crop coefficients for tobacco.
Crop coefficient
Weeks Zimbabwe1 Australia2 Australia2
after (early (late
planting planted planted
crop) crop)
4 0.11 0.50 0.53
5 0.20 0.59 0.62
6 0.40 0.66 0.71
7 0.65 0.72 0.78
8 0.90 0.76 0.84
9 1.00 0.80 0.88
10 1.00 0.83 0.92
11 1.00 0.85 0.94
12 1.00 0.85 0.95
13 0.80 0.85 0.95
14 0.70 0.83 0.93
 15 0.60 0.80 0.90
16 0.77 0.86
17 0.72 0.81
18 0.66 0.75
19 0.60 0.67
20 0.52 0.59
1 Tobacco Research Board (1988c).
2 Derived from Tonello, et al. (1993).

It is noticeable that from about 4 to 6 weeks after planting, the crop

coefficients of Zimbabwe are much lower than those of Australia,
despite the fact that a 4 to 5 weeks' stress is recommended after
planting in both countries. The slower growth in Zimbabwe may be
due to the very dry conditions which normally occur there.
Ideally, tobacco should be transplanted into soil which has been
brought to field capacity by rain or irrigation. The usual practice in
Zimbabwe is to plant the rain-fed crop with 2 to 5 litres of water per
plant about 2 to 3 weeks before the start of the rains. Goodall (1958)
reported that no root initiation took place in relatively dry soil
conditions, and this confirms the need to plant into a moist soil,
preferably at field capacity.

 Page 92
Papenfus and Quin (1984) divided the water requirements of a
tobacco crop into three phases. In the first, which is from 2 to 5 weeks
after transplanting, the plant is sensitive to over-watering and has a
small requirement for water. There are a number of reports of the
beneficial effects of early moisture stress, which encourages deeper
root development and avoids leaching of nitrogen. Consequently,
these findings indicate that in most circumstances watering should be
withheld for a few weeks after transplanting (Miles, 1957; Tobacco
Research Board, 1988c; Collins & Hawks, 1993; Tonello, et al.,
1993).
The second phase is from 5 weeks after planting through topping to
the stage when the upper leaves are fully expanded, usually about the
time that the third and fourth primed-leaves are harvested. This stage
is one of rapid growth and the crop needs frequent irrigation or
rainfall. Water stress during this period will decrease the yield and
quality of tobacco significantly (Collins & Hawks, 1993; Tobacco
Research Board, 1986d).
The third phase extends over the remainder of the primed-leaf
harvesting period, when the water used by the plant decreases
significantly because there are fewer leaves and the rate of
transpiration is reduced (Papenfus & Quin, 1984). Tobacco is more
tolerant to moisture stress during this stage and water is only
necessary to ensure the leaves ripen properly and cure satisfactorily.
However, severe moisture stress does reduce leaf quality (Papenfus &
Quin, 1984). Thus, irrigation is sometimes used on a supplementary
basis to offset the adverse effect of a dry period (Collins & Hawks,
1993; Reynolds & Rosa, 1995; Georgia Cooperative Extension
Service, 1996).
Response to Climatic Factors
The effects of cultural practices and some environmental factors, such
as rainfall and soil fertility, have been discussed in other sections. This
section concentrates on the effects of humidity, temperature, light and
CO2 on the growth, yield and quality of tobacco. Many other
climatological factors affect tobacco and these have been discussed by
Mulchi (1985) and Tso (1990e).
Excluding genetic factors, the potential yield and quality of tobacco is
the product of the interaction of the environment in which the tobacco
is grown and the cultural practices applied to the crop (Bertinuson, et
al., 1970; Court, 1982; Stocks, 1986). Attempts have been made to
develop models for the growth of tobacco by using mathematical
equations that mimic the physiological processes of photosynthesis,
respiration, carbon partitioning and leaf emergence. A simple model
includes climatic data of daily maximum and minimum air
temperature, thermal units which are calculated using a base threshold
of 10°C and photosynthetically active radiation (PAR) (Schneider &
Sturgill, 1994). Other models concentrate on photosynthesis using
functions of light, temperature and CO2 concentration (Pachepsky &
Acock, 1996). Climatic variables are not easily controlled in the field,
and interpretations of the effects of individual factors are complicated
by their interdependence and the diurnal fluctuations of most of them.
The influence of one variable, such as temperature, on the growth and
development of plants varies according to the intensity of the others.
Therefore, extrapolation of findings from controlled-environment
chambers to the field should be done with extreme caution. However,
Raper, et al. (1975) have attempted to define the conditions required
in controlled-environment rooms that simulate those in the field.
Long-term field studies in localities with different climates are
required in order to improve our understanding of the effects of
climate on the growth of tobacco. In addition, when collecting and
analyzing climatic data some thought should be given to the use of a
single variable, such as ET0. The ET0 determines the effect of climate
on crop-water requirements, and if calculated using the modified
Penman method, it incorporates humidity, temperature, solar radiation
and wind speed (FAO, 1977).
a
Humidity
Plants that are growing in environments with different relative
humidities exhibit variations in traits that are associated with the
regulation of water loss, particularly due to their cuticular and
stomatal properties. It is a common observation that best leaf growth
and expansion occurs when the evaporative demand is relatively low.
Yet few of these responses can be attributed solely to humidity and
none can be shown to be controlled by a 'humidity sensor' in the plant
(Grantz, 1990). Solar radiation, CO2 concentration of air, temperature
and sometimes atmospheric pollutants have an effect on stomatal
conductance and should be thought of when considering the effects of
humidity (Turner, 1969; Grantz, 1990; Lösch & Schulze, 1995).
Transpirational water diffusion through the stomatal pores is driven by
the leaf-to-air vapor pressure difference and restricted by the variable
diffusion resistance of the stomatal pores (Lösch & Schulze, 1995). A

 Page 93
high relative humidity is associated with a low vapor pressure deficit,
which results in lowered transpiration rates and keeps the stomata
open for gas exchange. As relative humidity decreases the rate of
transpiration increases initially and then decreases as the stomata
close. When this occurs, the water content of leaves generally
increases (Kappan, et al., 1995).
The decreased transpiration associated with high humidities may also
adversely affect the mass flow of nutrients from the roots to the leaves
(Long & Woltz, 1977). Raper and Smith (1975) found that leaf areas
of tobacco grown under a high relative humidity of 90% were larger
in the lower and midstalk positions and smaller in the upper stalk
positions than for plants grown at a lower relative humidity of 65%.
The decrease in leaf area of the upper leaves was a response to the
humidity during the first 3 to 5 weeks after transplanting. Initial
periods of high humidity, whether followed by a low or high humidity,
also resulted in abnormally elongated leaves. An initial period of low
humidity was thought to be important in ensuring adequate
concentrations of nitrogen in the tissues during leaf formation. As a
result, they proposed that the effects of humidity on leaf phenotype
would be attributed to the effects of nitrogen uptake by mass flow.
The accumulation of dry matter/unit leaf area, starch and alkaloids
was greater under high daytime humidities than low daytime
humidities, irrespective of the night humidity. The increased starch
and dry matter production by plants under high day humidity were
probably related to lower stomatal resistance and increased net
photosynthesis. It was suggested by Long and Woltz (1977) that the
higher alkaloid concentration was a result of the greater
photosynthetic capacity which increased root growth and, therefore,
alkaloid synthesis.
It must be emphasized that the work of Raper and Smith (1975) was
done in controlled-environment rooms, whereas, under field
conditions the adaptation to other factors, such as soil temperature and
light intensity, might be more significant than adjustment to humidity.
b
Temperature
Temperature is an important factor contributing to plant vigor. At
temperatures that are too high or too low the photosynthetic yields
decrease steadily until the uptake of CO2 ceases. After a small and
short-lived deviation from the favorable temperature range, the
photosynthetic function returns to the previous level of activity.
Exposure of plants to more extreme temperatures results in
perturbations that are not immediately reversible upon return to
optimum conditions. The after-effects of temperature stress are serious
structural and functional impairments that can only be restored to
normal by repair processes. Inhibitory temperatures depend on species
and genotype. Also, these temperatures are not constant but depend on
the stage of development, the state of activity of the plant and the
environmental conditions (Larcher, 1995).
In the field, overheating of the leaves may result from strong solar
radiation. The midday depression of photosynthesis on clear, hot days
is due, among other factors, to heat-related reversible photoinhibition.
A number of different species are able to raise their heat threshold for
photosynthetic functioning from morning to afternoon. The rise in
heat tolerance can be achieved within a few hours by synthesis of
polypeptides and heat shock proteins which protect the chloroplasts
(Larcher, 1995).
Low temperatures are much more damaging to plants in light than in
darkness. Chilling-sensitive C3 plants show photoinhibition of
photosynthesis when exposed to chilling temperatures. Typically,
chilling injury is manifested as an inhibition of the maximum quantum
yield of CO2 uptake and indicates an impaired photochemistry
(Öquist, et al., 1987). In chilling-injured plants, the rate of respiration
and ion leakage increases. The leaves may wilt on chilled plants,
particularly if they are not kept at high humidity. After plants have
been returned to nonchilling temperatures, leaves often appear water
soaked and necrotic lesions may occur. Although the chilling injury
may be reversed if the duration of the chilling period is sufficiently
short, the recovery in the post-chilling period is not immediate and
depressed rates of plant growth are commonly noted (Hetherington &
Öquist, 1988).
The temperature requirement of a particular plant species can be
expressed by a vegetative thermal constant which is the sum of the
average diurnal temperatures of the entire vegetative stage. The
constant for tobacco is 3200°C to 3600°C (Sebánek, 1992).
It has been shown, using constant day/night temperatures throughout
growth, that tobacco responds more to night temperatures than day
temperatures, and the optimum night temperature decreases with
increasing plant size and fluctuates around 20°C (Camus & Went,
1952; Haroon, et al., 1972; Long & Woltz, 1977). Despite these
findings, day temperatures and other factors, such as root
temperatures, are very important. The relatively low optimum
temperature for net photosynthesis in C3 species may be related to an
increase in respiration, a reduction in the diffusion

 Page 94
of CO2 and an increase in photorespiration as the daytime temperature
increases (Long & Woltz, 1977). Day temperature affects the
transpiration rate, which is generally greater at higher temperatures
because the stomata have a tendency to become wider (Kappen, et al.,
1995). However, excessively high temperatures result in stomatal
closure and a reduction in net photosynthesis (Long & Woltz, 1977).
Raper and Downs (1976) compared the growth of tobacco under
phytotron culture and field conditions. They found that in the
phytotron, singular thermoperiods (e.g. 26/22°C) did not produce a
reasonable copy of the field-grown phenotype and that low
temperatures resulted in elongated leaves and high ambient
temperatures produced broad leaves. When temperature schedules
simulated seasonal temperature progressions in a field environment,
they established that day temperature during the period of leaf
initiation was associated with the determination of leaf shape of
tobacco. They concluded that the field phenotype of a crop could be
reproduced in phytotrons that were capable of simulating seasonal
increases in temperatures and decreases in the availability of nitrogen
and potassium during growth.
In another study by Osmond and Raper (1981), root zone
temperatures had greater effects on growth than either aerial
temperature or nitrate concentration. There was a marked decrease in
shoot dry weight at root temperatures of 16°C compared to 24°C and
32°C. The reduction in growth at 16°C apparently was attributable to
initial water stress and the reduced loading of nitrogen from the root
into the vascular system. In addition, there was an interaction effect
among low solution nitrate concentration, low root temperature and
medium and high air temperature.
A direct correlation between temperature and the rate of carbohydrate
translocation has been observed in a number of plant species (Long &
Woltz, 1977). Wallace and Abou-Zamzam (1973) exposed the roots of
topped tobacco to air at temperatures between 14° and 23°C. The
amount of cations in the exudate of the xylem and the volume of the
exudate were positively correlated to temperature. If the temperature
were decreased there was also a lag in the appearance of the exudate.
Generally, total alkaloid levels are positively correlated with
increasing day/night temperatures (Long & Woltz, 1977). Raper and
Johnson (1971) compared day/night temperatures of 18/14, 22/18,
26/22 and 30/26°C and found that higher temperatures increased total
nitrogen and total alkaloid concentrations in the lower leaves and
decreased them in the upper leaves. The higher alkaloids in the lower
leaves were attributed to increased nitrogen accumulation and crop
duration. The 18/14°C regime increased the reducing sugar
concentration in the lower leaves. Potassium concentrations were
highest in the lower leaves at the lower temperatures and in the upper
leaves at higher temperatures. In the field, however, the potassium
concentration of tobacco leaves is normally highest in the lower
leaves.
In their review, Long and Woltz (1977) suggested that the higher
concentration of sugars in the lower leaves at low temperature might
reflect the reduction in translocation from those leaves at low
temperature. However, the effect of the lower nitrogen concentration
on the level of sugars should not be overlooked, as the accumulation
of carbohydrate is a typical response to internal nitrogen stress
(Weybrew, et al., 1983; Rideout, et al., 1992).
There is evidence that the activity of the enzyme sucrose-phosphate
synthase (SPS) is involved in the regulation of sucrose formation and
export. In their work on soybeans, Rufty, et al. (1985) found that SPS
activity decreased at low temperatures and they concluded that the
activity of SPS was an integral part of the coordinated adjustments in
the leaf photosynthetic system which limited the formation and export
of sucrose as the temperature declined.
The exposure of plants to low temperatures, particularly at night,
causes an accumulation of starch in a number of species. In cotton,
starch accumulated in warm night-grown plants (28°C) after exposure
to a relatively low temperature of 20°C for one night and was depleted
in cool night-grown plants (20°C) after only one night at 28°. This
quick response supports the idea of a reversible biochemical reaction
(Warner & Burke, 1993). Rideout, et al. (1992) reported that in
tobacco, a low temperature regime of 18/14°C caused an increase in
starch accumulation and a decrease in the percentage of total soluble
carbohydrate compared with 30/26°C. There was also an apparent
inequality in the relative concentrations of carbohydrate and nitrogen
in the apical meristem at the time of floral transition and they
considered that this supported a modified nutrient diversion
hypothesis for floral development as proposed by Raper, et al. (1988).
Severe freezing injury to relatively mature tobacco leaves resulted in a
decreased concentration in the cured leaves of reducing sugars and an
increased concentration of starch (Rosa, et al., 1984). Therefore, there
is sufficient evidence to show that temperatures affect carbohydrate
metabolism and translocation. However, it is difficult to generalize

 Page 95
about any one aspect, such as reducing sugars, as there are a number
of processes involved.
As tobacco plants grow they become increasingly 'ready to flower'
(Kasperbauer, 1969; Hopkinson & Ison, 1982) and it has been shown
that low temperature, particularly at night, accelerates the time of
transition to the floral stage in a number of cultivars (Camus & Went,
1952; Papenfus, 1980; Rideout, et al., 1992). However, the
environmental control of flowering in tobacco is complex and is
affected by temperature and photoperiod, both independently and in
combination (Garner & Allard, 1920; Hopkinson & Hannam, 1969;
Kasperbauer, 1969; Thomas, et al., 1975; McDaniel, 1992).
c
Light
Plants that are grown in dense populations are often taller and lighter
in color than those grown in sparse populations. One difference in the
growth environment between these two populations is light intensity,
which is affected by the extent of shading of one plant by another.
Spectral distribution of light received by shaded leaves in high
populations may also differ from that received by unshaded leaves.
Light of certain qualities, particularly red and far red, is known to
function in the regulation of the growth and development of the plant
(Kasperbauer, 1971). These responses are extremely sensitive to light,
often responding to very low intensities, and are controlled by the
phytochrome pigment. Phytochrome exists in two forms, Pr and Pfr,
which are reversible; the Pr form absorbs red light and changes to the
Pfr form and the Pfr form absorbs far-red light and switches to Pr.
Thus the red:far-red ratio of light determines the photoequilibrium
which regulates the developmental processes.
Kasperbauer (1971) determined that tobacco leaves shaded by other
leaves received more far-red light, relative to red, than was received
by unshaded leaves. It was also reported that high populations of
tobacco produced plants with elongated stems; however, the leaves
were slightly shorter and much narrower compared with leaves from
low populations. These results were probably due to a shift in the
spectral composition of light which was received during the
development of the plant. In another experiment under controlled
conditions, Kasperbauer (1971) investigated the developmental
responses to end-of-day red and far-red light. The number of leaves
produced per plant during an 18-day treatment period was not affected
by the end-of-day treatment. However, leaves on plants that received
the far-red light just before darkness were relatively narrow, light
green and had less dense laminae than those that received red light.
Red-irradiated plants had a greater dry mass of whole plant and roots,
greater root to shoot ratio and shorter stems than those receiving far-
red light. Suckers also grew in the axils of the lower leaves. Plants
that received red light last had higher concentrations of alkaloids and
free amino acids, and lower concentrations of soluble phenolics,
particularly chlorogenic acid, free sugars and organic acids.
Light intensity and duration are important in the growth of tobacco.
Chen and Huang (1970) reported that the accumulation of dry mass
and the dry mass/unit area increased with increasing light intensity
and duration. The ratio of dry to fresh mass increased as the light
period increased. Raper, et al. (1971) found that increased light
duration enhanced the relative width of leaves. Raper and Johnson
(1971) reported that the concentrations of potassium, nitrogen and
alkaloids were lower with increased light duration, although this may
have been a result of dilution from the greater dry mass. In contrast,
Tso, et al. (1970) noted that plants that received 16-hour photoperiods
had significantly higher concentrations of total alkaloids and total
phenolics than those that received only 8-hour photoperiods.
The saturation intensity of light for net photosynthesis in tobacco is
about 3000 foot candles, approximately 305 W/m2 (full spectrum) or
600 mmol/m2 PAR (conversions derived from Thimijan & Heins
(1983)). Shade-grown plants have a lower saturation intensity of about
1000 foot candles (200 mmol/m2 PAR) (Burnside & Böehning, 1957;
Hesketh & Moss, 1963; Moss, 1964). Delgado, et al. (1992) found in
field experiments that light was not limiting at about 1000 mmol/m2
PAR, and similar values were obtained by Calè and Rea (1991).
However, it should be remembered that the saturation intensity is
affected by temperature, humidity, moisture stress and CO2
concentration (Papenfus, 1970; Hall & Rao, 1987; Calé & Rea, 1991;
Pachepsky & Acock, 1996) and the different values reported are
probably due to the conditions under which the experiments were
done.
Most plants experience many fluctuations in sunlight from full-sun to
shade throughout the day. Many fist-growing herbaceous species
rapidly reduce stomatal opening during short-term shade periods.
Closure of the stomata during shade conserves water but may reduce
CO2 uptake and rate of photosynthesis (Knapp & Smith, 1990).
Turner and Incoll (1971) reported that, in tobacco, the penetration of
light and

 Page 96
rate of photosynthesis declined with depth in the canopy. The stomata
in the lower leaves of a well watered tobacco crop failed to open, even
in bright light. While the stomata of the mid-canopy leaves opened,
photosynthesis was less than that of the upper leaves at an equivalent
stomatal conductance. This may have been due to factors such as
shade decreasing the concentrations of chlorophyll and Rubisco or
limiting levels of CO2 in the crop canopy (Monteith, 1962; Long &
Woltz, 1977). Papenfus (1967) attributed the decreased capacity for
photosynthesis of tobacco leaves to be a combination of shading by
the upper leaves and aging. The relative importance of the aging
factor increased with time.
Fluctuations, particularly diurnal, in light and temperature have a
marked effect on the growth and development of tobacco in the field.
Long and Woltz (1977) found that the concentration of reducing
sugars in green leaf was high in the early morning hours and then
decreased and remained low. Starch concentration generally declined
during the day. Light can also have a great effect on the rate of uptake
of nitrate by roots, and decreased rates in darkness as compared with
light have been observed (Delhon, et al., 1995). Long and Woltz
(1977) emphasized the role that the diurnal fluctuations in light have
on plant metabolism and that the concentration of certain chemical
constituents and quality of cured tobacco may be influenced by the
time of day the leaf was harvested.
d
Carbon Dioxide and Climate Change
Carbon dioxide assimilated by tobacco plants is supplied from the air
above the crop and the respiratory activity of the leaves and soil.
Generally, the turbulence of the air at night is low, so that the CO2
gradients are rather steep and the highest concentration is near the
soil. Usually, the differences in CO2 concentration are smaller during
the day than at night, because air turbulence is greater. However,
photosynthetic CO2 uptake exceeds CO2 production by the soil and
plant respiration, and as a result the CO2 concentration is often lowest
in the top of the canopy where there is the most photosynthesis.
During the day, the CO2 concentration in the air above the crop is
highest early in morning and decreases during daylight hours.
Generally, CO2 concentration increases during the dark period
because of the respiration activity of plants at night and the lack of
photosynthesis (Monteith, 1962; Gaastra, 1963).
Raper and Downs (1973) and Raper, et al. (1973) compared the effect
of allowing plants to deplete the CO2 concentration of the air from
400 to 200 ppm in controlled-environment rooms with those grown in
an atmosphere in which the CO2 concentration was maintained at 400
ppm. Plants that were subjected to the low CO2 treatment had smaller
leaves and less dry matter accumulation in the shoots, but only
slightly lower starch, reducing sugar and polyphenol concentrations in
the cured leaves.
Respiration, photosynthesis and photorespiration occur
simultaneously in photosynthesizing cells. In leaves subjected to
moderate to high light intensity, at normal temperature and CO2
concentration, photosynthetic CO2 assimilation may proceed 10 to 20
times faster than CO2 release from respiration in the dark at the same
temperature (Amthor, 1995). However, plant species differ markedly
in their rates of photorespiration; in some inefficient species it may be
as high as 50% of net photosynthesis (Hall & Rao, 1987). A tobacco
mutant that exhibited O2-resistant photosynthesis was found to have
enhanced photosynthesis because the photorespiration was reduced
(Zelitch, 1990). However, the selection of tobacco lines for survival at
low CO2 concentration did not result in less photorespiration and
greater net photosynthesis, but rather they gave genotypes with
smaller cells which were able to use more photosynthate for cell
division and hence larger leaves were obtained (Delgado, et al., 1992).
The activities of humans release large quantities of atmospheric
pollutants including ozone, chlorofluorocarbons and oxides of carbon,
nitrogen and sulfur, which are often referred to as 'greenhouse gases',
and CO2 is one of the major components. The effect of ozone and acid
rain on tobacco growth was reviewed by Mulchi (1985). Hall and Rao
(1987) reported that the CO2 concentration of the atmosphere has
increased from 270 to 345 ppm in the span of a century and is
estimated to reach 600 ppm in 50 years.
The enzyme Rubisco is a catalyst in the fixation of CO2. At current
values in the atmosphere, the CO2 concentration is too low to achieve
saturation of Rubisco in C3 plants, such as tobacco, and O2 competes
for the reaction sites. Increasing atmospheric CO2 increases the
intercellular CO2/O2 ratio at the reaction sites of the Rubisco, and
decreases oxygenation and hence photorespiration. Therefore, there is
a positive response of net photosynthesis to high CO2 concentrations,
particularly at higher temperatures (Lawlor, 1995).
Most plants have a decreased stomatal conductance under elevated
CO2 values, and if this reduced the transpiration from all vegetation it
would have impor-

 Page 97
tant implications for crop water use and the hydro-logical cycle as
atmospheric CO2 continues to rise. However, several other
mechanisms tend to diminish the effect of the reduction of stomatal
conductance in terms of its effect on canopy evapotranspiration. A
major counterbalancing effect arises when leaf area/plant increases
due to growth under elevated CO2. However, Samarakoon, et al.
(1995) found that for wheat, a C3 species, this counterbalancing of
stomatally reduced transpiration/unit leaf area by greater leaf
area/plant applied only under drying soil conditions. Under
continuously wet conditions there was a minimal leaf area response to
CO2; thus, plants in the high CO2 environment had reduced
transpiration rates and used significantly less water. These authors
concluded that the relative growth enhancement by high CO2
concentrations increased with drought, and that the high CO2 greatly
relieved drought stress.
It was pointed out by Schulze and Caldwell (1995) that, despite all the
knowledge that has been collected about photosynthetic processes,
there is still considerable uncertainty in making predictions on
wholeplant performance at elevated CO2 due to other mechanisms
within the plants that might be affected.
With the current global climate changes, more studies on the effect of
elevated CO2 and temperature on the physiology, yield and quality of
tobacco are required, so that field practices may be altered to
maximize production.
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Chapter 5
Production Practices
5A
Flue-Cured Tobacco
G.F. Peedin
North Carolina State University
Raleigh, North Carolina, USA
Introduction
Tobacco is the most widely grown commercial nonfood crop in the
world (Capehart & Grice, 1994). Although cotton is grown on more
land area, tobacco is produced in at least 117 countries by
approximately 33 million farmers (Campbell, 1995a), primarily
between latitudes 45°N and 30°S (Tso, 1990). In 1993, total world
production of all classes of tobacco was estimated at 8.42 million
metric tons on 5.14 million hectares, less than 0.5% of the total
available arable and permanent croplands in the world. Approximately
5.39 million metric tons or 64% of total production in 1993 was
classified as flue-cured tobacco (Creek, et al., 1994).
Fluecured tobacco, also called Bright or Virginia, derives its name
from the unique curing process used to produce lemon to orange
colored cured leaves with high sugar contents and smooth smoking
properties. Curing is the process for drying freshly harvested tobacco
with partially or fully controlled temperature and moisture schedules
that preserve the inherent quality of the leaves during drying.
Fluecuring occurs in tightly constructed barns with artificial heat
beginning at about 35°C and ending at about 75°C over a 5 to 7 day
period. The term 'flue-cured', first used to describe Bright tobacco,
was derived from the system of metal flues used to distribute the heat
over the floor of the curing barn. The initial and most important stage
of flue-curing is called 'yellowing' during which chlorophyll is
degraded and most carbohydrates are converted to simple sugars. The
leaf cells are then killed with drier and hotter air to stop respiratory
losses of sugars. Most other tobacco types are primarily aircured
under prevailing environmental conditions over a period of 4 to 8
weeks, usually in well ventilated structures with little or no artificial
heat, resulting in cured leaves with relatively low sugar contents.
Fluecured tobacco has a characteristic sweet aroma due to its high
sugar content, and a slightly acidic taste. Because of these properties,
it blends well with other tobacco types (such as burley, Maryland and
Oriental) to produce a smooth and flavorful smoke in cigarettes.
The flue-curing process apparently originated in 1839 in Caswell
County, North Carolina, on a farm with sandy, infertile soils
considered practically worthless for production of other commodities.
In the early 1800s, farmers were attempting to produce a mild
flavored tobacco for pipe smoking, especially for consumers abroad.
It was thought that a thin, yellow cured tobacco would provide the
desired aromatic smoke, but such tobaccos were rare. In those days,
small fires were used on the floors of open curing barns to lower the
humidity and improve natural curing conditions. It seems a slave
responsible for curing fell asleep during the yellowing stage and
almost allowed the curing fire to go out (Collins & Hawks, 1993).
Because the woodpile was wet, he quickly rekindled the fire with
charcoal from the blacksmith's forge and, in his haste, raised the
temperature much higher than normal for tobacco curing at that time.
The result was bright, yellow cured leaves which sold for four times
the normal price! However, the flue-curing process did not become
widely used until after the American Civil War (1865) when cigarette
smoking became more popular and substantially increased demand for
thinner-bodied, lighter-colored leaf compared to the thicker-bodied,
harsher and darker aircured tobaccos then used primarily for plug and
twist chewing tobaccos. Cigarette smoking increased steadily but
slowly until the early 1900s when the cigarette-making machine was

 Page 105
developed and came into general use, making cigarette smoking less
expensive. More details on factors influencing the development of
flue-cured tobacco production in the southeastern USA are given by
Collins and Hawks (1993).
Today, approximately 80% of total world tobacco production is grown
for use in cigarettes, currently estimated at 5.6 trillion pieces annually,
up from 3.2 trillion pieces in 1972 (Stevens, et al., 1996). Flue-cured
tobacco is used primarily in cigarettes, although small amounts are
used in pipe and chewing products and as cigar filler. Cigarettes made
totally or primarily with flue-cured tobacco are quite popular in
China, Europe and Asia. Flue-cured tobacco is the major component
of the blended cigarette, first introduced in the USA by R.J. Reynolds
in 1913 (Collins & Hawks, 1993). World consumption of the
American blended cigarette is presently increasing about 5% per year
(Dale Hill, personal communication, Philip Morris USA), making the
USA the second leading cigarette manufacturing country in the world
(behind China) and the world leader in cigarette exports (Stevens, et
al., 1996). The estimated composition of the American blended
cigarette by tobacco type and source since the late 1960s is shown in
Table 5.1. The use of Maryland tobacco, which can be replaced in
blends by burley, has declined more than 50%, while total relative use
of flue-cured, burley and Oriental has not changed substantially.
However, relative use of US flue-cured (and burley) has declined
about 30% over the last 20 to 25 years, likely the result of better
tobacco processing and cigarette manufacturing technology and/or
changes in consumer preferences that allowed for greater use of less
expensive but usable tobaccos from other parts of the world. Notice,
however, that the relative use of both domestic flue-cured and burley
increased in 1994 to 1996 due to import restrictions imposed by the
1994 Domestic Content Law, which was considered inconsistent with
GATT/WTO requirements and replaced in 1995 with the less
restrictive Tariff Rate Quota System.
World production of flue-cured tobacco has more than doubled over
the last 30 years (Table 5.2). Although 80% of total production still
occurs in the same 8 to l0 major countries, absolute and relative
productions among these countries have changed considerably since
the late 1960s (Tobacco Associates, Inc, 1980, 1989, 1997). China
emerged as the leading flue-cured producer (and consumer) by far, but
is not currently a major exporter. Production in Brazil and Zimbabwe
increased 4.5 and 9 times, respectively, while that in Canada, Japan
and the USA declined significantly. During 1993 to 1996, Brazil and
Zimbabwe supplied nearly 45% of the world's flue-cured exports,
including over 70% of that imported into the USA; India and China
had approximately equal shares of the world export market, averaging
5 to 7% each (Tobacco Associates, Inc, 1997).
Production Practices
The amounts of specific cured tobaccos suitable for various cigarette
blends are determined primarily by their chemical and physical
properties, which in turn are the products of interactions among
genetic, soil, climatic and cultural factors. The inherent adaptability of
flue-cured tobacco allows it to be successfully grown in at least 75
countries, primarily between latitudes 60°N and 40°S (Collins &
Hawks, 1993). The seasonal rainfall and temperatures averaged over
several major production areas in the USA, Brazil and Zimbabwe, not
greatly dissimilar in latitude, are shown in Table 5.3. These climatic
factors are, on average, distinctly different among the production
areas, with Brazil
Table 5.1 Estimated use of tobacco types in US cigarettes based on
annual disappearance, 196996.
Tobacco Average annual disappearance for years (% of
total)1
Type Source 196971 197981 198191 199293 199496
Flue Domestic 46.9 36.4 31.2 32.2 37.1
 Imported 0.7 5.2 8.6 14.0 12.3
Burley Domestic 36.8 33.6 30.0 26.8 28.2
Imported 0.3 8.4 9.8 12.9 9.6
Oriental Imported 13.6 14.3 14.0 13.3 12.6
MarylandDomestic 1.8 1.6 1.2 0.9 0.7
1 Calculated from data in Capehart (1997).

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Table 5.2 Flue-cured tobacco production in the eight major producing
countries, 196596.
Average annual farm sales weight (1000 MT/yr) and % of world
production1
Country 196569 % 197577 % 198587 % 199496 %
Brazil 65 3.9 165 7.2 261 7.9 307 7.9
Canada 94 5.6 95 4.2 72 2.2 67 1.7
China 374 22.4 605 26.6 1696 51.0 2104 54.1
India 96 5.8 97 4.3 107 3.2 115 2.9
Japan 123 7.4 98 4.3 66 2.0 51 1.3
Korea 48 2.9 82 3.6 55 1.6 54 1.4
USA 496 29.7 584 25.6 323 9.7 387 9.9
Zimbabwe 74 4.4 92 4.0 116 3.5 190 4.9
WORLD 1672 100.0 2277 100.0 3325 100.0 3890 100.0
1 Calculated from data in Tobacco Associates, Inc Annual Reports (1980;
1989; 1997).

having much higher rainfall but lower minimum temperatures than the
other two countries during the first half of the production season.
Zimbabwe's early season is very dry and tobacco transplanted during
this period normally requires irrigation until the rains begin. Notice
also the relatively high maximum temperatures during the latter half
of the production season in Brazil and the wide differences between
maximum and minimum temperatures for Brazil during all of the
production season compared to the relatively small differences in
Zimbabwe. Average yields in these areas during 1984 to 1994 were
approximately 2480, 2284 and 1830 kg/ha for North Carolina (Miller,
et al., 1997), Zimbabwe (Tobacco Marketing Board, 1994), and Brazil
(Sindifumo, Brazilian Tobacco Industrial Association, unpublished),
respectively, and are likely reflective of differences in soils and
management practices as well as climatic differences. While the
Table 5.3 Average rainfall and temperature comparisons for major flue-
cured production regions in North Carolina, Brazil and Zimbabwe.1

Production season (months)

Flue-cured region 1 2 3 4 5 6
Rainfall (mm)
N. Carolina 89 102 111 128 119 101
Brazil 149 167 160 137 140 191
Zimbabwe 24 97 176 234 195 105
Maximum temp (°C)
N. Carolina 24.8 28.4 30.8 31.3 29.6 25.5
Brazil 28.0 30.0 34.0 36.0 36.5 37.0
Zimbabwe 29.1 27.1 25.7 25.4 25.2 25.6
Minimum temp (°C)
N. Carolina 10.6 15.0 18.1 19.1 17.2 12.1
Brazil 5.5 7.5 9.0 12.5 14.5 14.5
Zimbabwe 16.3 17.9 18.1 17.8 17.6 16.3
Max - min temp (°C)
N. Carolina 14.2 13.4 12.7 12.2 12.4 13.4
Brazil 22.5 22.5 25.0 23.5 22.0 22.5
Zimbabwe 12.8 9.2 7.6 7.6 7.6 9.3
1 Calculated from data in sources: NC (Epperson, et al., 1988);
Zimbabwe (Stocks, 1994); Brazil (unpublished, supplied by Universal
Leaf Tobacco Co, Inc, Santa Cruz). Data for NC and Zimbabwe are
averages for two or more major production areas for at least 30 years;
data for Brazil are averages for three major production areas for 11 years.

 Page 107
interactive effects of these and other environmental factors, such as
relative humidity, on tobacco yield and usability are largely unknown,
the result is that many styles of flue-cured tobacco are produced in the
world and various cultural practices are used to favorably influence
leaf usability characteristics in the regions (i.e. environments) where
the crop is grown. The intensity of specific practices often varies
within and among regions and countries, but nitrogen fertilization
(and sometimes soil moisture), plant density, harvesting and curing
are the major production factors influenced by growers within their
local conditions to produce profitable yields of tobaccos suitable for
use in cigarette products. The grower's degree of success is measured
by leaf quality and yield, cost of production and the consistency of
producing a usable crop year after year (Hawks, 1975).
Bowman, et al. (1984) assessed the relative contributions of breeding
and production technology to yield and quality improvements of flue-
cured tobacco in the USA from 1954 to 1.981. During this 27-year
period, he estimated that 32% of the average annual yield increase
(49.5 kg/ha) was due to varietal development and the other 68% to
changes in cultural practices such as improved soil fertility, use of
multipurpose fumigants, herbicides, chemical sucker control and
irrigation. The ability and/or need to adopt new technology varies
over the world but appears to be most rapid in the major tobacco-
producing countries. For example, average yields have increased
approximately 9, 15 and 21% in North Carolina, Brazil and
Zimbabwe, respectively, since the early 1980s, although use of
varieties developed in the USA is practically nil in Zimbabwe (Ann
Jack, personal communication, Tobacco Research Board) and is
declining in Brazil (Sergio Bremm, personal communication,
Profigen, Inc). Ultimately, however, the level of production and
usability of the local cured product in a given season are highly
dependent upon suitable weather conditions, particularly amount and
distribution of rainfall. The purpose of this chapter is to highlight and
discuss those agronomic field practices which research has shown to
contribute to the efficient production of usable flue-cured tobacco
under a range of soil and weather conditions.
Seedling Production and Transplanting
Tobacco is a transplanted crop and seedling production is similar in
most countries, except where greenhouses are widely used. In
conventional seedbed production, seedlings are started from tiny seeds
(about 12 000 per g) sown by hand on well prepared soil beds and
manually removed for transplanting to the field after reaching a height
of 15 to 20 cm. In tropical climates, seedling beds are usually covered
with dried plant materials to preserve soil moisture and reduce
disturbance of seeds or seedlings by heavy rains. In cooler climates,
seed beds are covered for frost and freeze protection with one of
several porous synthetic materials or cotton cheesecloth until
approximately a week before transplanting. The purpose of removing
the covering at this stage of growth is to acclimate (i.e. 'harden') the
seedlings before transplanting them to the harsher field environment.
Withholding water periodically during the last 2 to 3 weeks before
transplanting is used to harden transplants in some countries. The bed
soils are usually treated before seeding with methyl bromide, dazomet
or metam sodium to manage most weeds and soil-borne diseases and
insects (Miner & Worsham, 1990). Herbicides for supplemental grass
control are labeled in some countries (Smith, 1996b), but in regions
where labor is plentiful and inexpensive, weeds and grasses are often
removed from seedling beds by hand. Foliar insects and diseases are
usually managed with appropriate pesticides (Southern, 1996; Melton,
et al., 1996).
In the USA and Canada, seedlings are produced primarily in
greenhouses covered with plastic and glass, respectively. It was
estimated that 80 to 85% of the flue-cured transplants used in North
Carolina in 1997 were grown in plastic-covered float greenhouses,
and their use is increasing in countries such as Canada, Italy and
Australia. Smaller, plastic-covered but less expensive outdoor
waterbeds, usually with a means of supplemental heat, are sometimes
used on small farms. Overhead-watered systems are less popular than
float systems in the USA because more management time and
capability are required for fertilization and irrigation, particularly
from seed germination until the root systems become established. In
Canadian glasshouses, regular seeds are broadcast on steam-sterilized
muckbased media, and the seedlings are then managed much like
those in conventional plant beds. In the float system, a peat-based
medium is contained in polystyrene trays with an individual cell for
each of several hundred seedlings. The trays are floated on fertilized
water and each cell is continuously moistened by capillarity. The
seeds are usually coated (i.e. pelletized) to increase their size, which
facilitates mechanical seeding, although pelletization is not necessary
for some types of seeders. The trays are often treated for disease

 Page 108
control with a sodium hypochlorite solution or preferably methyl
bromide before each transplant production season (Melton, et al.,
1996). Currently, only a few pesticides are labeled for use in tobacco
transplant greenhouses, so proper ventilation, horizontal air
movement, solarization (between production seasons) and good
sanitation in and around the greenhouse are used to help reduce
problems with foliar/stem diseases and insects.
Frequent clippings after the seedlings are 6 to 7 cm tall and allowing
night temperatures to drop to about 15°C in the latter 2 to 3 weeks of
growth are the primary means of hardening transplants in float and
overhead-watered greenhouses. In addition to clipping, both higher
greenhouse temperatures and root disturbance (i.e. forking), which
cause moderate wilting of seedlings, are used in various combinations
to harden seedlings in Canadian glass-covered houses (Van Hooren &
Veinot, 1997). Clipping is widely practised on conventional seedbeds
in some countries and its other advantages include improving
transplant uniformity/usability and slowing seedling growth when
needed. These are discussed further in Chapter 4B, Seedling
Production. Reed (1996) has published excellent guidelines on
clipping and other management practices for successful transplant
production in float greenhouses.
Depending on availability and cost of labor and equipment, seedlings
are manually or mechanically transplanted to well prepared fields
previously treated with one or more pesticides for control of soil pests
and/or weeds. Additional foliar pest management is usually needed
during subsequent growth and harvesting of the crop. (The specific
pesticides and application methods for target pests are discussed in
other chapters of this monograph.) However, to protect workers from
exposure, applications of hazardous pesticides near or during
transplanting are not recommended. In the USA, the recently enacted
EPA Worker Protection Standard (WPS) requires farmers to protect
workers from pesticide-related illness or injury by:
(1) providing training on pesticide safety, specifically for those
pesticides used on the farm,
(2) providing personal protective equipment and clothing and
assuming responsibility for their proper use and cleaning,
(3) ensuring that workers do not enter treated fields within specific
time intervals after pesticide application, and
(4) providing decontamination sites and emergency assistance in case
of exposure.
The WPS requirements for most flue-cured tobacco pesticides labeled
in the USA as of 1996 have been summarized by Moore and Southern
(1996). In many countries, varietal tolerances and multi-year rotations
of tobacco with nonhost crops (when sufficient land is available) are
widely used to reduce reliance on pesticides. In Zimbabwe, the dates
of seeding plant beds, transplanting to the field and the destruction of
plant beds and stalks/roots in harvested fields are controlled by
legislation to reduce the incidence and spread of insect-transmitted
viruses (Tobacco Research Board, 1988). In areas where appropriate
equipment is available, high row ridges are often formed several days
or weeks before transplanting. Compared to fiat soil, water infiltration
into ridges is reduced and soil temperature in the ridge at transplanting
is usually higher. When row fumigation is used on shallow soils,
injecting the fumigant into the soil during ridging allows a greater soil
volume to be treated than is usually possible when the fumigant is
injected into flat soil. Rapid root growth soon after transplanting is
desirable, but is reduced by excess moisture and/or low temperatures
in the root zone. Therefore, transplanting on ridges often improves
early plant growth in cool, wet conditions.
Most of the row ridge is pushed off during mechanical transplanting,
so the tobacco is transplanted on a lower, wider ridge. Depending on
the yield goal and style of tobacco to be produced, rows are spaced 90
to 122 cm apart and plants within rows 45 to 60 cm apart. The
transplants undergo a period of 'transplant shock', which lasts 5 to 8
days for conventional transplants, depending on soil moisture and
ambient factors affecting evapotranspiration. Good quality and
properly clipped transplants from float or overhead-watered
greenhouses usually undergo less wilting and recover more rapidly
than conventional transplants.
Water is normally applied manually or mechanically around the
transplant roots during transplanting to improve soil-root contact and
thus plant livability. The amount of water used per transplant ranges
from 100 ml to 12 L, depending on soil type and moisture content and
climatic conditions at the time of transplanting. In the USA, liquid
fertilizers with high phosphorus (P) concentrations are occasionally
added to the transplant water. Early growth is sometimes stimulated
and plants may flower several days earlier than normal, even on soils
with high P availability, but yield or leaf quality are not improved
(Smith, 1995; Smith, 1996a). Early growth responses are similar for
transplants grown in greenhouses or plant beds when

 Page 109
the liquid fertilizers are used at rates to provide 4 to 8 kg/ha of P2O5
(phosphorus pentoxide). In field experiments, however, excessive
fertilizer concentrations in the transplant water sometimes stunted
transplants from plant beds more than those grown in float
greenhouses (Fisher, 1997; G.A. Pate, MS research in progress, NC
State University). Therefore, the organic media surrounding much of
the root mass of greenhouse transplants may provide some protection
against fertilizer salts injury. It has been reported that root and shoot
growth of greenhouse float transplants are sometimes slower than that
of transplants grown in conventional plantbeds, particularly when
weather conditions for the first several weeks after transplanting are
unusually dry (Fisher, 1997) or cool (Pate, MS research in progress,
NC State University).
Cultivation and Weed Management
Hawks (1975) stated that tobacco cultivation serves three major
purposes:
(1) reduces weed growth,
(2) improves soil aeration and water penetration, and
(3) can be used to form a large row ridge (called 'ridge cultivation')
which diverts some excess water to row middles and thus reduces the
risks of drowning and loss of leachable nutrients.
Compared to flat cultivation, Collins, et al. (1968) obtained yield
increases with ridge cultivation which were greater at wet locations
than at normal or dry locations in North Carolina. Collins and Hawks
(1993) suggested that the comparatively lower moisture content in the
ridge under wet conditions may contribute to higher temperatures in
the root zone which improves root growth. In addition, it is generally
accepted that roots initiate on stalks covered by the soil provided by
ridge cultivation. Collectively, the additional soil and root mass
reduce lodging and increase absorption of water and nutrients.
Cultivation is the major method of weed control for flue-cured
tobacco. In Zimbabwe, where labor is relatively plentiful and
inexpensive, manual cultivation is routinely practiced, even for ridge
formation after the crop is established. Mechanical ridge cultivation is
practiced in countries where production units are large and cultivating
equipment is available. Within a few days after the transplants recover
from transplant shock, the first of two to four cultivations is made and
a portion of the fertilizer nutrients is applied. With each cultivation,
soil is moved to the plants from the sides of the rows, partially
reforming the row ridge lost at transplanting by moving increasing
amounts of soil to the plant stalks. Cultivation is usually completed
within the first month after transplanting, resulting in an increase in
ridge height of 10 to 15 cm. The majority of the ridge reformation
occurs during the last or 'layby' cultivation when the plants are 30 to
35 cm tall.
The number and frequency of cultivations needed on the same soil
may vary from year to year, depending on the weed population, soil
type and frequency and intensity of rains. Soils that crust badly at the
surface after drying may need to be cultivated after each rain to
improve aeration. The level of weed infestation, often higher with
frequent rains after transplanting, and whether a herbicide has been
used, will also determine when cultivations are needed. Tobacco is a
shallow rooted crop and most of the roots, even at maturity, are in the
upper 30 to 40 cm of soil. Cultivating deeper, more often, or later than
needed to achieve the major purposes of cultivation may injure roots
which reduces water and nutrient uptake and increases the risk of
infection by soil-borne diseases such as bacterial wilt (Pseudomonas
solanacearum) and black shank (Phytopthora parasitica) (Smith,
1996b). Collins and Hawks (1993) suggested that as the plants
become larger and their root systems more extensive, any soil
disturbance closer than 8 to 10 cm to the plants should become
progressively more shallow. For the last 'layby' cultivation, when the
primary buildup of the ridge occurs, most of the soil moved to the
plants should come from the middles between rows. Also, mechanical
cultivation after plants infected with tobacco mosaic virus are tall
enough to touch the tractor can spread the virus over the field.
Weeds may reduce cured leaf yield and quality directly by competing
with tobacco for growth resources or indirectly reduce quality as
foreign matter in the cured product. Therefore, herbicides are often
used to complement the weed control provided by cultivation. Their
use allows more flexibility in scheduling cultivations and may reduce
the number of cultivations needed, but is unlikely to replace the need
for cultivation on many flue-cured soils. Research in North Carolina
(Hawks & Collins, 1970) indicated that when a herbicide was used,
yield was not reduced when the number of cultivations was reduced
from the normal 35 to 2, but yield was significantly reduced when
only one cultivation was used. Use of the herbicide and no cultivation
resulted in the lowest yield and value per ha. In another study, Collins,
et al. (1972) imposed two or no cultivations on three herbicide
treatments.

 Page 110
Compared to the nontreated, twice-cultivated control, cultivation of
the herbicide treatments increased yield an average of 5%, while not
cultivating the same herbicide treatments reduced yield 10%.
Herbicide and cultivation treatments did not affect the average market
price of cured leaves.
Successful weed management includes proper use of other cultural
practices in addition to cultivation and herbicide use (Smith, 1996b).
Early stalk and root destruction destroys most weeds before their
seeds mature, reducing weed populations the next season. Herbicides
labeled for tobacco usually will not control all weeds present in
tobacco fields. In addition, use of the same herbicides on the same
land year-after-year may increase populations of resistant weeds.
Therefore, rotation of tobacco with other crops, which also aids in
disease management, allows the opportunity to use a wider range of
herbicides which may be more effective on problem weeds. This is
particularly true for some broadleaf weeds which are not adequately
controlled by most tobacco herbicides (e.g. Xanthium strumarium,
Ambrosia artemisiifolia and Cassia obtusifolia). A fast growing
tobacco crop aids in weed control by shading the row ridges and
middles. Therefore, practices which encourage healthy root systems
(use of healthy transplants, crop rotation, disease control,
transplanting on ridges, proper rate and application methods of
pesticides and fertilizers, and maintaining proper soil pH) are all part
of a sound, integrated weed management program (Collins & Hawks,
1993).
Herbicides are used sparingly in some major flue-cured countries such
as China, India and Canada. In the USA, however, approximately
80% of the land area is treated with one or more herbicides. The
relative degrees of weed control for the herbicides currently labeled
for use on flue-cured tobacco in the USA are shown in Tables 5.4
(grasses) and 5.5 (broadleaf weeds). Diphenamid and isopropalin are
no longer registered in the USA but are legal to use when available.
Postemergence grass herbicides such as sethoxydim and fluazifop-
butyl have given good-to-excellent control of certain grass species
(Hagwood & Komm, 1984) but are not yet labeled. Some weeds differ
in susceptibility to herbicides, so accurate field records of weed
species and populations are necessary for proper selection and rate of
herbicides (Smith, 1996b). Pendimethalin, pebulate and napropamide
control weeds by restricting cell division during seed germination.
The other two compounds are absorbed by emerging roots and shoots
and affect photosynthesis (clomazone) or plant metabolism
(sulfentrazone) and it is not uncommon to observe susceptible weeds
emerge before their growth is controlled. More details on the activity
and use of these herbicides is given in Chapter 5B on air-cured
tobacco.
Sulfentrazone received a federal label for use on tobacco in the USA
in early 1997. It is a member of the aryl triazolinone group, classified
as a protoporphyinogen oxidase inhibitor, and acts by the same
mechanism as the diphenyl esters through membrane disruption. Soil
mobility is greater than that of alachlor but less than that of metribuzin
(FMC Corporation, 1993). Sulfentrazone provides better control of
Cyperus (nutsedge) species than pebulate (Table 5.4) and is the only
tobacco herbicide labeled in the USA that controls Ipomoea (morning
glory) species (Table 5.5). However, it is relatively weak on several
grass species (Table 5.4) and tankmixing with another herbicide such
as clomazone is necessary to achieve broad spectrum grass control.
Most weed seeds germinate in the upper 2 to 3 cm of soil. Certain of
the above herbicides can be soil incorporated (PPI) or applied to the
soil surface (PRE-T) before transplanting, applied over the crop
within the first week after transplanting (OT) or applied soon after the
lay-by cultivation (LB). There are advantages and disadvantages to
each time and method of application, depending on weather
conditions, weed populations and the herbicide used. In the USA, all
of the herbicides except sulfentrazone are labeled for PPI application.
Advantages include:
(1) more consistent weed control than surface applications because
herbicide incorporation and activation are less dependent on rainfall
or irrigation,
(2) tankmixing with certain other soil-applied pesticides to save one or
more passes over the field (Walls, et al., 1974; Johnson, et al., 1992),
and
(3) it provides weed control when the first cultivation is delayed by
excess rainfall.
The primary disadvantages of PPI applications are stunting of tobacco
and/or following grass crops. Stunting of tobacco is more likely
during cool, wet conditions, particularly on coarse, low organic matter
soils when the herbicide is applied at excessive rates, improperly
incorporated or tankmixed with another herbicide. Reduced rates of
PPI grass herbicides often provide acceptable control but less stunting
in fields treated with herbicides for several years. Although
clomazone has not been observed to stunt tobacco when properly
applied, some occasional whitening of leaves may occur but maturity
and yield are not affected (Smith, 1996b). Napropamide (LaMondia &
Ahrens, 1996), some dinitroanalines (Shelby, et al.,

 Page 111
Table 5.4 Relative grass control with herbicides labeled for flue-cured tobacco
in the USA1 (Smith & Fisher, 1998).
Weeds PendimethalinSulfentrazonePebulateNapropamideClomazone
Barnyardgrass G-E F G-E G-E E
(Echinochloa
crus-galli)
Bermudagrass P P P P P-F
(Cynodon
dactylon)
Broadleaf G F-G P G E
Signalgrass
(Brachiaria
platyphylla)
Large Crabgrass E F-G E E E
(Digitaria
sanguinalis)
Crowfootgrass E F E E E
(Dactyloctenium
aegyptium)
Fall Panicum G F-G G G E
(Panicum
dichotomiflorum)
Yellow Foxtail E F-G E E E
(Setaria glauca)
Goosegrass E F-G G E E
(Eleusine indica)
Johnsongrass G F G F G
(seedlings)
(Sorghum
halepense)
Field Sandbur G P-F G
(Cenchrus
incertus)
Texas Panicum G F P G
(Panicurn
texanurn)
Nutsedge P E G P P
(Cyperus
esculentus)
(Cyperus
rotundus)
1 Ratings based on adequate soil and weather conditions and on proper rate,
method and timing of application.
E = excellent, >90%; G = good, 8090%; F = fair, 6080%; P = poor, <60%.

1990; LaMondia & Ahrens, 1996) and clomazone (Smith, 1996b)

applied PPI may stunt small grains following tobacco. Soil residue
effects on fall-seeded wheat following burley tobacco were enhanced
when pendimethlin and flumetralin were used for weed and sucker
control, respectively, on the tobacco; stunting of tobacco planted the
following spring was due primarily to flumetralin carryover (Shelby,
et al., 1990). Also, because of the potential for soil residue carryover
with sulfentrazone, there is an 18-month planting restriction for cotton
(Gossypium hirsutum) and 12-month restrictions for several other
crops. (FMC Inc has applied for a PPI label for sulfentrazone in 1998
because weed control is more consistent than with the PRE-T method
currently labeled. However, considerable stunting and necrosis of
tobacco has sometimes been observed with sulfentrazone applied with
any of the methods listed above, particularly with PPI and OT
applications.)
Surface applications of herbicides (PRE-T and OT) usually do not
provide acceptable weed control as consistently as PPI applications
and generally are recommended for fields with a history of low-to-
moderate grass pressure. However, surface applications are often
adequate in formerly high-pressure fields which have been treated for
several years with PPI herbicides while in tobacco and other crops.
The less volatile, micro-encapsulated (ME) clomazone formulation
can be applied on the soil surface before transplanting, and this
application method was required for sulfentrazone in the USA in
1997. If ridged rows are used, they should be pushed down to

 Page 112
Table 5.5 Relative broadleaf weed control with herbicides labeled for flue-
cured tobacco in the USA1 (Smith & Fisher, 1998).
Weeds PendimethalinSulfentrazonePebulateNapropamideClomazone
Common P F-G P P F
Cocklebur
(Xanthium
strumarium)
Common E G-E G E F-G
Putslane
(Portulaca
oleracea)
Hairy P F-G P P-F P-F
Galinsoga
(Galinsoga
ciliata)
Jimsonweed P F-G P P G
(Datura
stramonium)
Common G E G G G
Lambsquarters
(Chenopodium
albumm)
Morning P E P P P
Glories
(Ipomoea sp.)
Redroot G-E E G G P
Pigweed
(Amaranthus
retroflexus)
Prickly sida P G-E P P E
(Sida spinosa)
Common P P P F G
Ragweed
(Ambrosia
artemisiifolia
Sicklepod P P P P P
(Cassia
obtusifolia)
Pennsylvania P-F E P P G
Smartweed
(Polygonurn
pensylvanicum)
1 Ratings based on adequate soil and weather conditions and on proper rate,
method, and timing of application.
E = excellent, >90%; G = good, 8090%; F = fair, 6080%; P = poor, <60%.

transplanting height and the herbicides applied as close to

transplanting time as Worker Protection Standards allow.
Napropamide and clomazone can be applied over the crop within a
week after transplanting. Both application methods are dependent on
soil moisture to activate and move the herbicides into the soil and thus
fit well in overhead irrigated situations. If rainfall does not occur
within 3 to 5 days after application, a light cultivation often improves
weed control. The primary advantages of PRE-T and OT surface
applications are that the risk of stunting tobacco and following crops
is reduced and the total amount of herbicide applied can be reduced by
using narrow bands over the row, where weeds are more difficult to
control with cultivation, rather than broadcasting it over the entire
field. However, because sulfentrazone is relatively mobile, substantial
rain on coarse-textured, low organic matter soils can move it into the
root zone, causing stunting and necrosis of tobacco as mentioned
above. In addition, re-ridging of fields treated with surface-applied
herbicides such as sulfentrazone or napropamide concentrates the
herbicide in the root zone and can cause significant injury to tobacco
(Smith & Fisher, 1998).
Lay-by applications are useful in extending the period of grass control
in high-pressure fields when a short-lived herbicide such as pebulate
is used, particularly in wet seasons. A lay-by application of
pendimethalin or napropamide following a PPI application of pebulate
will cause less stunting of tobacco than a PPI tankmix of pebulate
with one of these products. However, pendimethalin application to
tobacco buds will cause injury and should be avoided. Yield
reductions do not occur unless weed populations are unusually heavy,
so the use of a lay-by herbicide should be considered on a year-to-year
basis or when the use of mechanical harvesters necessitates excellent
weed control to minimize foreign matter in the cured leaves. Potential
herbicides and other pesticides undergo

 Page 113
strenuous residue and smoke panel evaluations before being approved
for use on tobacco in the USA. Therefore, their use as labeled would
not be expected to negatively influence the smoking properties of
cured leaves.
Field Soils, Fertilization and Soil Moisture
a
Soil Properties and Nutrient Disorders
Flue-cured tobacco is typically grown on sandy, infertile surface soils
with good internal drainage and low organic matter contents (i.e. low
nitrogen reserves) to enhance the light-color and high sugar
characteristics of cured leaves. Most air-cured tobaccos, except
Maryland and Oriental, are usually grown on heavier-textured, more
fertile soils which also receive substantially higher nitrogen (N) rates.
Some characteristics of both flue-cured and burley tobacco surface
soils in North Carolina are shown in Table 5.6. The generally higher
sand content of flue-cured soils is indicated by the higher percentage
of soils with high volume weights and low cation exchange capacities
(CEC). Humic matter content is also lower in flue-cured soils. Humic
matter represents that portion of organic matter that has decomposed
into humic and fulvic acids and is not necessarily indicative of the
total organic matter content of the soil. In North Carolina, flue-cured
soils with humic matter contents greater than about 0.8% often
produce tobacco that ripens slowly and is difficult to cure properly
because N is still being absorbed during the harvesting season, even
when fertilizer N rates are reduced substantially (personal
observation). The yield, quality and chemical composition of flue-
cured tobacco are directly associated with the development and health
of the root systems, which are sensitive to prolonged periods of excess
soil moisture. High humic matter contents can be indicative of soils
with poor internal drainage, which produce relatively good yields in
dry seasons but extremely poor yields in wet seasons. Soil acidity and
available calcium (Ca) are almost always adequate for optimum yield
and quality when soil pH is maintained in the 5.8 to 6.0 range with
dolomitic limestone (Peedin & McCants, 1977a). However, soil pH
less than 5.0 may result in aluminum (Al) and/or manganese (Mn)
toxicity to roots, while pH greater than 6.2 can cause deficiencies of
some micronutrients on soils with inherently low. availabilities of
these nutrients (Kamprath & Foy, 1985). On coarse, deep sands in
very wet
Table 5.6 Some properties of flue-cured and
burley surface soils in North Carolina.
Soil Property Flue-Cured Burley
property range (coastal plain) (mountains)
% of total samples1
Volume 0.750.94 1 26
weight
(g/cm3) 0.95- 69 73
1.29
>1 .29 30 1
Humic 0.250.50 42 34
matter
(%) 0.500.75 23 19
>0.75 30 40
Buffer <1 62 25
acidity
(meq/100 11.9 32 42
cm3)
22.9 4 28
pH 5.05.4 20 23
5.55.9 47 31
6.06.4 27 24
P (mg/kg) 3160 6 16
61120 21 30
>120 71 34
CEC2 <2.9 43 1
(meq/100 35.9 54 32
cm3)
69.9 3 57
K 0.130.25 37 21
(meq/100
cm3)
0.260.50 49 48
>0.50 6 27
Mg (% of <4.9 1 1
CEC)
59.9 21 8
1014.9 40 17
1519.9 28 24
Ca (% of <34.9 11 11
CEC)
3544.9 25 18
4554.9 37 35
5564.9 21 31
1Derived from North Carolina Department of
Agriculture soil test summaries for 1994 and
1995 crop years. Total number of soil samples
= 15 808 for flue-cured and 1261 for burley.
2 CEC = summation of buffer acidity; K, Mg,
and Ca expressed as meq/100 cm3.

early seasons in North Carolina, magnesium (Mg) deficiency is

occasionally observed in the second or third year after dolomitic lime
application (Peedin, 1996), but the condition does not occur in seasons
of more normal rainfall if available Mg is greater than 0.5 meq/100
cm3 of soil and occupies at least 10% of the cation exchange capacity
(M.R. Tucker, personal communication, NC Department of
Agriculture).


 Page 114
The data in Table 5.6 represent soil test availabilities of some nutrients
in a wide range of surface soils planted to tobacco for many years and
should not be interpreted to necessarily represent those nutrient levels
needed for optimum yield and cured leaf quality or to prevent certain
nutrient disorders. For example, fertilizer phosphorus (P) application
on soils with >60 mg/kg of extractable P was found to occasionally
increase early growth of tobacco, but did not increase yield and
quality (Lolas, et al., 1978) or substantially influence chemical
composition of cured leaves (Lolas, et al., 1979). In a 3-year study
conducted at 13 locations in North Carolina, Denton, et al. (1987)
found that the highest optimum rates of applied potassium (K) for
flue-cured tobacco ranged from 0 to 112 kg K2O/ha and the highest
rates occurred on soils with sand or loamy sand surfaces. However,
the K level in the Ap soil horizon was not related to yield response.
These authors suggested that leached K accumulated in the upper part
of the B horizon, not normally included in soil samples, and was more
available to roots on soils with thin, loamy surfaces than on soils with
deep, sandy surfaces.
Weather and/or genetic factors can also influence expression of
deficiency or toxicity symptoms of some nutrients. Calcium
deficiency symptoms occur before flowering on some flue-cured and
burley varieties, but not on others grown in the same experiments
following periods of rapid growth. The symptoms on flue-cured
tobacco were not related to available soil Ca or corrected by soil
applications of calcium sulfate and/or dolomitic limestone (Peedin &
McCants, 1977b). Work with burley tobacco indicated that the genetic
condition influencing Ca deficiency symptom development may
induce higher concentrations of oxalic acid in the upper portions of
susceptible varieties which complex Ca and reduce its translocation to
and/or metabolic accessibility in apical meristems (Brumagen & Hiatt,
1966). The inability to maintain adequate Ca in meristemic tissues is
apparently more pronounced when rates of upper leaf expansion are
unusually high. Using controlled environmental chambers, Rufty, et
al. (1979) found that tolerance of tobacco to high rates of applied Mn
increased with increasing temperature, despite greater concentrations
of Mn in green leaves during early growth. They proposed that the
greater tolerance to Mn in warm temperatures is associated with more
rapid leaf expansion rates accompanied by greater vacuolor capacity
for disposal of accumulated Mn. Leaching of sulfur (S) can cause
deficiencies of this nutrient on sands and loamy sands with clay
depths greater than 35 to 40 cm (Smith, et al., 1987). In addition, the
soil test levels for S in leachable surface soils may vary substantially
within and among growing seasons, depending on when the samples
are taken relative to leaching rains. Thus, the soil test for S, like that
for K, is generally unreliable for predicting optimum application rates.
For most of the disorders discussed above, tissue as well as soil
analysis is often helpful in diagnosing and correcting the nutrient
problems. An excellent review of most nutrient disorders for tobacco,
the sufficiency ranges in leaf tissues and proper sampling procedures
was published by Miner and Tucker (1990). Color photographs and
descriptions of most nutrient disorders on tobacco are also available
(Reich, 1991).
b
Micronutrients
The essential micronutrients for nonluguminous plants are boron (B),
chloride (Cl), iron (Fe), manganese (Mn), molybdenum (Mo), copper
(Cu) and zinc (Zn). Except for Cl, which is discussed later, tobacco
has relatively low requirements for micronutrients (Collins & Hawks,
1993) and deficiencies have not been sufficiently documented under
field conditions to warrant their general inclusion in flue-cured
tobacco fertilizers in the southeastern USA. However, Mo deficiencies
related to low pH and high sulfate applications have been reported for
burley tobacco in Kentucky (Miner & Sims, 1983) and for Maryland
tobacco (Khan, et al., 1994). Conversely, Ryding (1991) concluded
that Mo deficiency is not a problem for burley or flue-cured tobacco
in Zimbabwe on soils maintained above pH 5.0 (determined in 0.01M
CaC12) with lime.
Boron deficiency is the most common micronutrient deficiency of
flue-cured tobacco. It has been reported following dry conditions in
Virginia (Jones & Leslie, 1986) and in Zimbabwe (Ryding, 1986)
where B application is routinely recommended for tobacco production
(Tobacco Research Board, 1988). In the Mareeba-Dimbulah District
of Australia, B deficiency is associated with high rainfall during the
preceding wet season and is prevented by B application at
transplanting or corrected by one to three foliar sprays during the first
3 weeks after transplanting (Tonello & Gilbert, 1990). These workers
warned that there is a fine line between B deficiency and toxicity, and
no more than 1.25 kg B/ha over the 3-week period is recommended in
Australia.
Manganese deficiency sometimes occurs on flue-cured tobacco grown
in overlimed soils of the North Carolina coastal plain (Peedin, 1996).
In 10 on-farm problem situations investigated 4 to 7 weeks after

 Page 115
transplanting, average soil pH in the row was 6.35, available soil Mn
averaged 6 kg/ha and the average Mn concentration of flecked, green
lower leaves was 10 ppm (Peedin, unpublished data). In Georgia, mild
flecking symptoms on lower green leaves occurred in the first year of
a 2-year study when soil pH after harvest was 7.2, no fertilizer Mn
had been applied to limed plots and composited cured leaves averaged
13 ppm Mn (Stephenson, et al., 1987). Available soil Mn levels were
not reported in that study. Miner, et al. (1987) found that one to three
foliar sprays of 0.45% Mn between 3 and 7 weeks after transplanting
increased Mn concentrations of green leaves, but that band application
of manganese sulfate was six-fold more efficient than broadcast
application and was more effective in increasing Mn concentrations of
middle and upper leaves on a soil with pH 6.4 and 5.4 kg/ha of
extractable Mn. However, Mn deficiency symptoms did not develop
in these experiments, and yield and quality were not affected by Mn
rates and application methods. Jones and Leslie (1986) found no yield
and quality response of flue-cured tobacco to band or broadcast B, Cu
and Zn applied singularly or in various combinations in 13
experiments conducted on Virginia soils that tested low for these
micronutrients. Boron and Zn concentrations of 20 ppm and a Cu
concentration of 10 ppm in upper leaves at early flowering were
considered sufficient for flue-cured production in Virginia. However,
B toxicity occurred under dry conditions with as little as 0.56 kg/ha of
band-applied B.
Some knowledge of nutrient mobility within plants is often helpful in
diagnosing deficiencies. For example, N, Mg and Mn are very mobile
and move from lower to upper leaves when soil supplies of these
nutrients are inadequate to support growth. Therefore, deficiencies of
these nutrients are first visible on lower leaves. Conversely, nutrients
such as S, Ca, and B are not sufficiently mobilized from lower to
upper leaves when root absorption is inadequate, so deficiencies of
these nutrients are first observed on young upper leaves and buds.
c
Nitrogen, Vegetative Growth and Moisture
A number of excellent review articles describe the effects of macro-
and micronutrients on the agronomic, chemical and physical
properties of flue-cured tobacco (McCants & Woltz, 1967; Elliot,
1974; Chaplin & Miner, 1980; Miner & Sims, 1983; Sims, 1985; Tso,
1990). However, many tobacco soils have been in production for
years and contain substantial levels of the less leachable nutrients
within rooting depth. Failure to apply these nutrients annually at
traditional rates seldom limits yield and quality when soil pH is
maintained in the desirable range. Further, when a specific nutrient is
limiting, routine soil testing with improved extraction and correlation
techniques provides reliable rate estimates for most nutrients needed
for profitable crop production. However, N is leachable, and soil tests
for it are not reliable. Nitrogen is usually the most limiting nutrient in
the sandy, low organic matter soils preferred for flue-cured
production. More importantly, the ability to control both the amount
and time of N availability during growth and ripening, coupled with
moisture supply, has a greater effect on the agronomic and smoking
properties of tobacco than any other nutrient or management factor.
The association of low nicotine and high sugars in well watered
tobaccos and, conversely, of high nicotine and low sugars in drought
stressed tobaccos is well known. Weybrew and Woltz (1974) stated
that droughts are metabolically equivalent to overfertilization with N
while excessive leaching of N is equivalent to inadequate N
fertilization because the balance between N and carbohydrate
metabolism is greatly influenced by these production factors.
Weybrew's explanation is demonstrated by the data in Table 5.7
(Weybrew, et al., 1983) and is covered in more detail in Chapter 4C,
Field Practices, by Flower. When both N fertilization and soil
moisture are adequate for the timely uptake and reduction of N in
green leaves, the transition to starch accumulation occurs at about the
same time as flowering; leaves fill out, ripen properly and cure easily.
The cured leaves are compositionally balanced with respect to sugars
and nicotine (sugar/nicotine ratio between 6 and 8). According to
Weybrew, et al. (1983), when these two constituents are in proper
balance, it can be reasonably assumed that other chemical constituents
are not grossly unbalanced. Therefore, chemically balanced tobaccos
produce a smooth, flavorful smoke. However, excess N and/or
drought delay the transition from N reduction to starch accumulation,
allowing for more nicotine synthesis and, because of shortening day
lengths, less starch accumulation; these cured tobaccos are chemically
imbalanced, with sugar/nicotine ratios less than 5, and produce a
harsh, irritating smoke. Conversely, inadequate N and/or leaching
cause starch accumulation to occur prematurely, thereby restricting
nicotine production and permitting greater starch accumulation; these
cured tobaccos are imbalanced chemically, with sugar/nicotine ratios
greater than 9, and produce a flat,

 Page 116
Table 5.7 Main effects of N rate and soil moisture on some agronomic
and chemical properties of flue-cured tobacco (Weybrew, et al., 1983).
Main Yield Grade Total N Nicotine Reducing Sugar/nicotine
effect (kg/ha) index (%) (%) sugars (%) ratio
N
(kg/ha)
43 3093 23.9 2.03 3.22 21.6 7.1
64 3085 20.4 2.23 3.88 17.0 4.8
86 3013 13.9 2.62 4.60 12.5 3.0
Rain + irrigation
(mm)1
508 3392 24.3 2.01 3.23 21.3 7.1
406 2963 17.4 2.38 4.24 16.7 4.4
330 2836 16.3 2.49 4.23 13.1 3.4
1 Totals for 140-day production period.

insipid smoke. More recent work by Flower (1996) in Zimbabwe

generally confirms the basic explanation proposed by Weybrew, et al.
(1983), but indicates that a decline in leaf expansion and therefore N
demand, an increase in root growth and nicotine synthesis due to
topping, and a reduction in water use by the plant at this time may
also reduce N concentration and reduction in green tobacco leaves.
Other adverse effects of inadequate or excess N on the agronomic
properties and chemistry of flue-cured tobacco have been summarized
by Collins and Hawks (1993) and by Flower in Chapter 4C of this
monograph. It is generally accepted that proper ripening and curing of
flue-cured tobacco are enhanced when the soil N supply becomes
essentially exhausted soon after expansion of upper leaves is complete
(McCants & Woltz, 1967; Weybrew, et al., 1983). Therefore, only 50
to 90 kg N/ha are usually applied to flue-cured soils in most countries
(Dale Hill, personal communication, Philip Morris USA), depending
on the soil sand content, depth to clay, and the amount and distribution
of seasonal rainfall. In the southeastern USA, where leaching of N is
likely all year round on the sandy-textured tobacco soils, fertilizer N
recommendations are not based on soil analysis but on results of
numerous field experiments conducted over a range of soil types and
rainfall conditions. In North Carolina, the suggested N rate range from
routine fertilizer applications, assuming no leaching losses, is 55 to 90
kg N/ha, based primarily on topsoil depth, with modifications made
for the variety being grown and possible residual N left in the soil by
rotational crops (Peedin, 1996). The lower portion of the N rate range
is suggested for fine-textured, fertile soils, especially when legume
crops such as soybeans or peanuts were grown in that particular field
the previous year. The higher portion of the range is suggested for
coarse-textured soils with topsoil depths greater than 35 to 40 cm.
Although this approach performs well in seasons of normal rainfall,
adjustments for leaching are sometimes necessary in seasons of
excessive rainfall, particularly in areas with deep, sandy soils. Based
on field experience and the work of Terry and McCants (1970), an
adjustment procedure was developed by Hawks and Collins (1978)
that estimates the amount of additional N to apply based on topsoil
depth, the estimated amount of water that percolates through the root
zone, and the age of the crop when leaching occurred. This subjective
procedure, or a modification of it, is used effectively in many
countries around the world. Slow-release fertilizers reduce leaching
but are more expensive than water-soluble nutrient sources. Their
release rates are often too slow to supply N and other nutrients at
levels needed by tobacco during the period of rapid leaf expansion,
and the residual soil N supply remaining after topping is unpredictable
(Miner, et al., 1978; Chaplin & Miner, 1980; Miner & Sims, 1983).
The subjective aspect of N management for tobacco has yet to be
eliminated. However, Lyons, et al. (1996) evaluated sap analysis as an
aid in determining the need for additional N application to flue-cured
tobacco grown in Australia. The nitrate concentration in sap from the
midrib of the youngest fully expanded leaf (YFEL) quickly showed
the need for extra N before visual symptoms of N deficiency appeared
and in time for effective remedial action. Total N analysis of dried
green leaves was less effective for indicating additional N
requirements. These authors concluded that sap nitrate concentrations
in YFEL midribs should be greater than 5000 mg/L at final
cultivation, greater than 3000 mg/L at flowering, but less than 2500
mg/L at the start of harvesting for optimum yield and quality.

 Page 117
However, the decision to apply additional N would depend on soil
depth and the judgment that low nitrate values were due to leaching of
N out of the root zone and not to reduced N absorption by roots due to
waterlogging (i.e. oxygen starvation).
Tobacco has a relatively short period of leaf expansion which limits
the time for application of N and other nutrients. The time between
transplant recovery and flowering is about 8 weeks in suitable
environments, and it is usually during this period that nutrient
deficiencies occur (Miner & Tucker, 1990). Raper and McCants
(1966) characterized the accumulation of dry matter and nutrients in
the aboveground portions of flue-cured tobacco during a growing
season in North Carolina. Based on percentages of the totals, dry
matter accumulation increased at a lower rate than that of the nutrients
until flowering. Approximately 60% of the N, 45% of the K and Mg
and 37% of the P and Ca were absorbed during the 3-week period
following transplant recovery, but only 20% of the total dry matter
accumulated during this period. By flowering time, however, about
75% of the dry matter had accumulated and almost all of the N and K
had been absorbed whereas accumulation of Ca, Mg and P continued
until the youngest leaves reached physiological maturity (see also Fig.
4.1, with data expressed as kg/ha rather than %). While these
accumulation rates would be expected to vary for different soils
and/or climates, the data illustrate that flue-cured tobacco has a high
demand for nutrients during the leaf expansion period and requires
adequate and readily available levels, particularly of N (McCants &
Long, 1971), early in the growing season to reach maximum growth
and quality potential. As a result, all planned fertilizer nutrient
applications are usually made within the first 3 to 4 weeks after
transplanting in most countries, usually in two applications or more on
leachable soils. The first application occurs at or very near
transplanting and provides 50 to 60% of the total N. The last
application occurs at or before the final lay-by cultivation.
Additional nutrients, particularly N, are sometimes applied between
the last cultivation and topping if rainfall is excessive during this
period. While research and observations have shown late N
application to be of limited value on most North Carolina soils,
particularly when applied foliarly (Peedin & McCants, 1973; Peedin,
1985), research in Zimbabwe indicated that in under-fertilized
situations and for some varieties a relatively small amount of soil-
applied N at or soon after topping improved the color and balance of
the nitrogenous and carbohydrate constituents in cured leaves without
greatly affecting ripening rate (Tobacco Research Board, 1994). In
Canada, Court, et al. (1996) reported that application of one half of
the total N at transplanting and the remainder from 4 to 10 weeks after
transplanting had little effect on the agronomic and chemical
characteristics of cured leaves. However, these results were obtained
on soils which usually require only 55 kg N/ha for normal production,
and in the first year of the 2-year study rainfall was lower than normal
for most of the growing season.
d
Form of N
The form of N needed by flue-cured tobacco, i.e. nitrate (NO3) and/or
ammonium (NH4), has been the subject of considerable research and
discussion since the late 1950s. While nitrate-nitrogen (NO3-N)
performs well in many situations, it is more expensive and more
leachable than NH4-N during the first month after application, often
requiring leaching adjustments on sandy soils in wet seasons.
However, the work by McCants, et al. (1959); McCants (1960);
McCants & Woltz (1963, 1967) and others (Gous, et al., 1971;
Rhoads, 1972) in the USA indicated that NH4-N often reduced
tobacco growth in greenhouse and field studies, except when
excessive rainfall may have caused relatively greater leaching of
NO3-N in several field experiments (Elliot, 1970). This led to the
recommendation that N-P-K fertilizers contain at least 30% of the
total N as NO3-N. In actual practice, the fertilizer NO3-N content
unofficially adopted was nearer to 50% and has since become widely
accepted in the USA and many other flue-cured producing countries.
However, most of the early field work compared ammonium sulfate
with sodium nitrate and all of the N was usually applied in a single
application, a practice seldom used on sandy soils. In addition,
ammonium sulfate produces a highly acidic reaction in and around the
fertilizer band which could reduce bacterial conversion of NH4-N to
NO3-N (nitrification) in the soil, especially when all of the N is
applied in one application. Therefore, Williams and Miner (1982)
compared sodium nitrate with urea, which raises pH and nitrifies
relatively fast, both with and without soil fumigation which also slows
nitrification. Based on soil, agronomic and cured leaf sugar and
alkaloid data collected from eight experiments over 3 years, they
found that nitrification of urea was complete within 4 to 6 weeks after
application and that urea was equivalent to sodium nitrate as an N
source for tobacco in five of eight experiments conducted under
conditions of adequate soil moisture, even on fumigated soils. Urea
was superior to sodium nitrate in one experiment where

 Page 118
leaching occurred within 2 to 3 weeks after fertilizer application.
However, use of urea reduced yield an average of 7% in two
experiments conducted under very dry conditions, which apparently
slowed the nitrification process. The results with urea under
conditions of adequate soil moisture were later confirmed in
Zimbabwe (Tobacco Research Board, 1986).
Collectively, these studies indicate that NH4-N sources such as urea
and ammonium nitrate can be more widely used to reduce fertilizer
costs and leaching of N without reducing leaf yield or usability when
dependable water supplies and adequate irrigation systems are
available. However, the ability to maintain soil pH in the range of 5.5
to 6.5 must also be considered, particularly for urea. Low soil pH
reduces nitrification rate (Cao, et al., 1991) as well as NH4+
absorption by tobacco roots (Tolley-Henry & Raper, 1989).
Conversely, the use of urea on soils with pH levels of 7 or higher
could increase N losses by volatilization (Cao, et al., 1992).
In the southeastern USA, where no more than 50% of the crop is
routinely irrigated, it is standard practice to apply a sidedress fertilizer
containing 100% NO3-N 1 to 2 weeks after application of the N-P-K
fertilizer. However, Peedin (unpublished data, Table 5.8) found that N
form in the sidedress fertilizer did not affect yield, visual quality or
concentrations of reducing sugars and total alkaloids of cured leaves
when the previously-applied N-P-K fertilizer contained 50% NO3-N.
Soil moisture was generally adequate and soil pH ranged between 5.5
and 6.2 among locations. The sidedressings were band-applied 10 to
12 cm deep and supplied 40 to 50% of the total fertilizer N, depending
upon location. The lack of sidedresser × location interactions indicates
that the results are valid for a range of soil types in temperate climates
when soil moisture and pH are maintained at desirable levels. These
results were later confirmed in six similar experiments conducted over
2 years in eastern North Carolina (Smith, 1993). However, at one
location, urea increased reducing sugar concentrations of upper cured
leaves about 10%, possibly because of the lower energy required for
the reduction and subsequent utilization of NH4-N (Court & Hendel,
1986).
e
N Form and Chloride Interactions
McCants and Woltz (1967) summarized several experiments which
indicated that NH4-N increases chloride (Cl) toxicity symptoms, that
high Cl concentrations reduce visual quality and combustibility of
cured leaves, but that Cl uptake and/or concentration in tobacco are
inversely related to pH. This is additional evidence that successful use
of NH4-N is at least partially dependent on maintaining the proper soil
pH, particularly in production areas where soils and/or
Table 5.8 Effects of N form in sidedress fertilizers on some agronomic
and chemical properties of flue-cured tobacco grown in North Carolina,
1981 and 1982 Seasons (G.F. Peedin, unpublished data).
Fertilizer N Grade Reducing Total
Sidedress
fertilizer as NO3 (%) Yield index1 sugars (%) alkaloids Total
(kg/ha) (%) N (%)
Average of 15 Average of 5 experiments
experiments
Calcium nitrate 100 2788 36 14.8 2.52 2.44
(15.5-0-0)
Sodium nitrate 100 2707 36 15.2 2.52 2.40
(16-0-0)
Ammonium 50 2784 36 15.0 2.62 2.46
nitrate (34-0-0)
Ammonium 0 2757 36 16.4 2.35 2.27
sulfate (21-0-0)
Urea 0 2769 36 16.1 2.52 2.37
(46-0-0)
LSD (0.10) NS NS NS NS NS
CV (%) 6 11 16 15 10
1 Grade index is a 199 rating based on official USDA grades; higher
values indicate better quality.

 Page 119
irrigation waters contain enough Cl to cause concentrations in cured
leaves to exceed 1% (Miner &Sims, 1983). There is little evidence
from field experiments, however, that N form increases Cl
concentration in cured leaves of tobacco grown on relatively low Cl
soils such as those in the southeastern USA. Warren (1990) conducted
three field experiments to evaluate possible interactive effects of four
N sources (ammonium sulfate, urea, ammonium nitrate and sodium
nitrate) and four Cl rates (8, 37, 75 and 112 kg/ha from KCl). At two
locations, Cl toxicity symptoms developed only on the lower four to
five leaves of plants fertilized with ammonium sulfate or urea, but
they occurred at lower Cl rates when ammonium sulfate was the N
source. Symptoms did not develop on plants fertilized with
ammonium or sodium nitrate, even though Cl concentrations were
often as high as those in lower leaves of plants fertilized with
ammonium sulfate or urea. Further, higher leaf Cl concentrations were
associated with the 100% NH4-N sources only for 35-day-old plants
fertilized with 112 kg Cl/ha. Chloride concentrations of green lower
and upper leaves sampled 70 days after fertilizer application or of
lower and upper cured leaves were not affected by N form. Williams
and Miner (1982) found that urea application on fumigated soil
resulted in substantially higher Cl concentrations than sodium nitrate
in tobacco leaves sampled 4 and 6 weeks after transplanting, but Cl
concentrations were similar in leaves of all treatments 2 weeks later
when NH4-N was nitrified to low levels in the soil. Cured leaves
contained less than 0.5% Cl irrespective of N source or soil
fumigation treatment, although the fumigants increased Cl
concentrations as much as 50% over those in the nonfumigated
tobaccos. Conse-quently, the results of both Williams and Miner
(1982) and Warren (1990) indicate that NH4-N sources can increase
Cl toxicity and/or Cl levels in young tobacco plants exposed to high
rates of Cl, but are unlikely to substantially increase Cl concentrations
of cured leaves if nitrification is complete by 4 to 6 weeks after
fertilizer application.
It is generally accepted in the southeastern USA that the application of
25 to 35 kg Cl/ha often improves the yield and sometimes the visual
appearance of flue-cured tobacco, but that higher rates often severely
reduce leaf quality with no further increase in yield (Peele, et al.,
1960; Neas, 1961; Sierra, 1966; McCants & Woltz, 1967). The effects
of Cl on the hygroscopic properties of cured tobacco leaves and the
associated adverse affect on their burning properties are wellknown
(Elliot & Vickery, 1961; Sierra, 1966). Chloride concentration in
cured leaves increases linearly as the amount of Cl supplied by
fertilizers increases (Tsai, 1979; Miner & Sims, 1983; Warren, 1990).
In addition, some soils and irrigation waters, particularly those in
coastal regions, contain substantial amounts of Cl. Sierra (1966)
showed that Cl concentration is higher in lower than upper cured
leaves, and Warren (1990) showed that rate of Cl concentration
increase was about four times greater in lower than upper leaves as Cl
rate increased from 8 to 112 kg/ha (Fig. 5.1). Warren's (1990)
regression equation for lower leaves was y = 0.014x + 0.556 (r2 =
0.99), where y = percent Cl in cured leaves and x = kg/ha applied
fertilizer Cl; the y intercept, 0.556, represents the Cl contributions
from sources other than the fertilizer, such as Cl-containing soil
minerals, irrigation water and fumigants.
 Fig. 5.1
Relationship between rates of fertilizer Cl and Cl concentrations in lower and
upper flue-cured tobacco leaves (Warren, 1990).
If the generally accepted maximum Cl concentration of 1% is based
on lower leaves, Warren's equation can be used to predict the
maximum Cl rate allowed to keep leaf Cl concentrations from
exceeding 1%. Therefore, if y = 1% Cl, the corresponding Cl rate is
32 kg/ha for lower leaves. If the same approach is used for upper
leaves, the acceptable Cl rate would be 120 kg/ha, but this rate would
substantially increase the risk of leaf quality reduction. The 32 kg
Cl/ha rate is in the 25 to 35 kg/ha rate range recommended in the
major flue-cured states in the southeastern USA (Collins & Hawks,
1993), where the maximum Cl content of tobacco fertilizers is
officially regulated and monitored by state Departments of
Agriculture (North Carolina Department of Agriculture, 1986).
It should be reemphasized that the y intercept of the regression
equation will vary somewhat, depending upon amount and
distribution of rainfall, soil types and other cultural practices such as
fumigation. Therefore,

 Page 120
the equations developed in one country or region may not apply
directly to another. In North Carolina, it is suggested that as little
fertilizer Cl as possible be applied on fumigated soils (Peedin, 1996)
because recommended rates of the most popular fumigants supply 30
to 80 kg/ha Cl, a portion of which remains in the soil and is available
for absorption by roots.
f
Time and Method of Nutrient Application
Miner and Sims (1983) suggested that only one-half to two-thirds as
much of a fertilizer nutrient is required to produce optimum crop
yields when the fertilizer is banded compared to broadcast application.
While the magnitude of response to banding can vary widely, greatest
benefits occur in:
(1) soils with very low to low soil test levels of the nutrient in
question, particularly if it is not leachable,
(2) cool climates, and
(3) acidic or alkaline soils.
Collins and Hawks (1993) reported results of a large number of N-P-
K fertilizer application on-farm tests conducted in North Carolina
over an 8-year period. Banding the fertilizer to one or both sides of the
row at transplanting or within 10 days after transplanting produced
more uniform growth and 3.5% higher yields than broadcasting or
banding the fertilizer in the ridge before transplanting. These authors
suggested that pre-transplant application methods allowed more time
for leaching losses on sandy soils in wet seasons and contributed to
fertilizer salts injury in dry seasons, particularly banding in the ridge
where the fertilizer may be less than 8 to 10 cm beneath the plant
roots. In these tests, the fertilizer bands at or after transplanting were
placed about 12 cm to the side of the plant row and 10 to 12 cm deep;
this is the recommended method for banded fertilizer applications to
flue-cured tobacco in North Carolina. Miner and Sims (1983),
however, reported that some farmers banded the N-P-K fertilizer on
the surface of the row and used shallow incorporation with rolling
cultivators, resulting in slow early growth in dry seasons. Batten &
Peedin (1993) applied the N-P-K fertilizer:
(1) broadcast 1 week before transplanting,
(2) in two shallow (1 to 2 cm) bands 1 week after transplanting, and
(3) in two deep (12 to 13 cm) bands one week after transplanting.
Deep fertilizer placement produced higher yields than shallow
banding or broadcasting in a season when very little rainfall occurred
during the first 6 weeks after transplanting. In a second test where
excessive rainfall occurred in the first month after transplanting, yield
was not affected by fertilizer application method or depth, but
broadcast application reduced grade index, average market price and
value/ha because of a higher proportion of pale, immature cured
tobacco. Total N in cured leaves was also reduced, indicating that the
broadcasting method did not provide adequate N in close proximity to
roots under the wet conditions. Miner and Sims (1983) suggested that
shallow or surface banding may result in N deficiency if rainfall is
inadequate for several weeks after fertilization, and that P remains
near the soil surface and is not positionally available to new roots.
Collectively, these results indicate that banding of the N-P-K fertilizer
at the depth of the root system at or within 10 days after transplanting
would be expected to produce good yields of high quality cured leaves
more consistently over a range of soil types and rainfall conditions
than the shallow band or broadcast methods.
Studies with placement depth of sidedress fertilizers such as sodium,
ammonium or calcium nitrate have not been encountered in the
literature, but it would be expected that deep placement would be less
critical for early growth because these fertilizers do not contain P and
have relatively high water solubilities. However, it is possible that
intense rains could erode the exposed fertilizers off the ridge and carry
N from the field in surface water.
Topping and Sucker Control
Tobacco is a flowering plant with a central, terminal meristem which
suppresses growth of axillary buds (suckers) by hormonal action until
the meristem begins to produce flowers. Topping, or removal of the
terminal inflorescence, stimulates root growth and slows the rate of
decline in net photosynthesis in remaining leaves, thereby diverting
growth resources into leaf rather than seed production which increases
size, yield and quality of cured leaves, particularly at the upper node
positions (Papenfus, 1987). Topping and improved sucker control
increase the levels of most of the neutral volatile compounds
associated with desirable smoke character (Weeks & Sehmann, 1986).
However, topping also hastens rapid and profuse growth of suckers
(Decker & Seltmann, 1971) which must be removed periodically by
hand, a costly and

 Page 121
disagreeable chore, or their growth controlled chemically to maintain
the yield and quality increases achieved by topping. Flowers are
removed manually or mechanically, primarily in the USA, and sucker
growth is retarded in most countries with applications of contact
and/or systemic growth regulators.
a
Time of Topping
The beneficial effects of topping decrease as the time of topping is
delayed and seeds are allowed to mature (Papenfus, 1970). Therefore,
time of topping has a pronounced influence on the yield and chemical
composition of cured leaves. Marshall and Seltmann (1964a) found
that the average yield reduction in North Carolina as topping was
delayed beyond the button stage was 16 kg/ha per day for hand
suckered plants and 30 kg/ha per day for plants treated with maleic
hydrazide, a systemic growth regulator which allows much less sucker
growth and increases yield compared to normal hand suckering
(Marshall & Seltmann, 1964b). Additionally, both sugar and nicotine
concentrations of cured leaves decreased with delay in topping. Elliot
(1966) found that delaying topping about one week later than normal
for Canadian tobacco generally reduced length and width of upper
leaves and lowered total alkaloids, but did not lower reducing sugars
in cured leaves. The general response to late topping was reduced
yield, but the effect was seasonal and not consistent for some of the
five varieties used in the 3-year study. In a later study, Elliot (1975)
found more definitive yield increases and greater upper leaf densities
with topping in the pre-button (when the seventeenth or eighteenth
leaf was about 5 cm in length) or button stages. All of the above
studies were conducted in the 1960s and early 1970s when average
yields were in the 2000 to 2250 kg/ha range; similar studies with more
recent, higher-yielding varieties grown with improved cultural
practices have not been encountered in the literature. However,
Collins and Hawks (1993) listed several other advantages of early
topping:
(1) early topping requires less labor and topping is completed before
the labor-intensive harvest season begins,
(2) early topped plants are more resistant to lodging by wind, which
reduces efficacy of contact and contact-local systemic sucker control
chemicals as well as harvesting efficiency
(3) early topping alleviates some of the growth stresses associated
with both too little and too uch soil moisture, and
(4) early topping reduces the attractiveness of plants to egg-laying
moths of several harmful insects.
Therefore, early removal of tops may reduce the cost of insect
management and possibly the undesirable residues and health hazards
associated with some insecticides.
b
Modes of Action and Effects of suckercides
Steffens (1979) reviewed the modes of action of tobacco growth
regulators and their effects on the chemical properties and residue
levels of cured tobacco in 1979; very little new chemistry for tobacco
sucker control has been developed since that time. Depending on their
mode of action, tobacco suckercides fall into three categories: fatty
alcohol contacts, contact-local systemics or systemics.
Fatty Alcohol Contacts
These products are widely used and are composed primarily of
straight chain n-octanol and n-deconal mixtures or only n-deconals.
To be effective, they must wet the sucker buds directly and should be
applied by methods that encourage their drainage down the stalks and
into leaf axils. Therefore, the best sucker control is obtained when
contacts are applied directly over the top of upright plants; application
in the button stage is more effective than in later flowering stages
because suckers are less developed. Also, when topping is delayed,
the uppermost leaves are larger and easily blown over the stalk if
winds occur during application; the folded leaves protect the stalk tops
from the spray and disrupt the spray pattern. Seltmann (1994)
described how the unique spiral arrangement of leaves on the tobacco
plant affects the flow of fatty alcohols down the stalk and thereby
influences the effectiveness of contact suckercides in the upper leaf
axils. The tobacco plant has a 3/8 phyllotaxy, which means that when
counting consecutive leaves downward on a topped plant, the stalk is
circled three times until the ninth leaf is directly below the first leaf. A
fatty alcohol emulsion was applied to the first, second and third upper
leaf axils in all possible combinations. When the emulsion was
applied only to the first axil, the third axil was missed; if applied only
to the second axil, the first and fourth axils were missed; and if
applied only to the third axil, then the first, second and fourth axils
were missed. Seltmann (1994) concluded that wetting of the top three
axils is needed for excellent sucker control

 Page 122
with contacts, but that greater volumes of emulsion reduced the
number of suckers missed below the axil of application.
Fatty alcohols disrupt the semipermeable properties of cell
membranes and cause rapid desiccation of cell contents of meristemic
tissues (Wheeler, et al., 1991). Therefore, contact concentration is
important and must be just high enough to affect the cell membranes
of the more tender, succulent sucker buds but not those of the tougher
leaf and stalk tissues. Consequently, the fatty alcohol concentration
needed for tobacco grown under dry conditions may be too high and
cause phytotoxicity on upper leaves of tobacco grown with ample
moisture, particularly if applied under conditions of low humidity,
high temperature and high pressure. Conversely, the lower
concentration needed to affect sucker buds without causing
phytotoxicity on succulent tobacco may not give sufficient effect on
buds of tobacco grown under dry conditions. The use of fatty alcohol
contacts as the only suckercide seldom gives satisfactory sucker
control because sucker buds not wetted by the initial application may
grow too large to be adequately controlled by applications made 2 to 3
days later (Collins, et al., 1970). Also, the contact concentration
needed to desiccate the meristemic tissues of the two to three buds
normally found in each of the upper axils of flue-cured tobacco
(Seltmann & Kim, 1964) could cause leaf phytotoxicity and possibly
leaf drop. In Canada, however, where the length of the harvest season
is relatively short and use of the systemic (maleic hydrazide) is
prohibited, several applications of fatty alcohols are the predominant
means of chemical control (Rosa & Caughill, 1991). Other than
possible phytotoxicity, the use of fatty alcohols is not known to
adversely affect normal development of immature leaves (Rosa &
Caughill, 1991), cause undesirable residues (Tancogne, et al., 1977),
or to substantially alter the chemical, physical and smoking properties
of cured leaves compared to manually-suckered tobacco (Steffens,
1979). Therefore, when early topping is practised to increase yield,
one or more fatty alcohol contact applications are frequently used to
provide sucker control until the upper leaves become large enough to
be treated with the systemic or contact-local systemic, both of which
can restrict growth of leaves less than about 25 cm in length.
Contact-Local Systemics
Another group of compounds now used worldwide for tobacco sucker
control are called 'contact-local systemics'. They are not translocated
within the plant but are absorbed sufficiently by meristemic tissues to
stop cell division in leaf axils contacted directly. Therefore, like the
fatty alcohols, contact-local systemics are most effective when applied
by manual or mechanical methods which encourage stalk rundown
and wetting of leaf axils rather than leaf absorption. Because these
chemicals inhibit cell division, however, application to immature,
upper leaves may cause growth restrictions, and generally, they should
not be applied as early as fatty alcohols (Collins & Hawks, 1993).
The most widely used contact-local systemics are dinitroanalines
(flumetralin, butralin, pendimethalin), although chlorothal-dimethyl
and chloropham are popular in some countries. The physical and
chemical properties of cured leaves from plants treated with
dinitroanilines compare favorably with those treated with sequential
applications of fatty alcohols and maleic hydrazide (Steffens, 1979;
unpublished data of the Regional Tobacco Growth Regulator
Committee). However, unless applied to individual plants by hand,
single applications of dinitroanalines seldom provide adequate sucker
control because fewer axils are wetted with machine than hand
application (Peedin, 1987). Using manually-applied sprays, Jones and
Rideout (1986) found that tankmixing flumetralin with a 4% fatty
alcohol improved sucker control and yield compared to flumetralin
application alone. Sucker control was improved for both treatments
when applied after rather than before topping, primarily for the low
rate of flumetralin (0.84 kilograms active ingredient/hectare (kg ai/ha)
vs 1.35 kg ai/ha). Yelverton, et al. (1993) used butralin alone
following one or two fatty alcohol applications in seven experiments
conducted across the flue-cured states in the southeastern USA. They
reported that butralin can be an effective tobacco suckercide, but
multiple applications generally were more effective than single
applications and directed downstalk application provided better
control more consistently than over-top foliar applications. These
authors concluded that missed suckers are a problem for
dinitroanilines and their use should be evaluated in combination with
the systemic, maleic hydrazide.
Dinitroanilines are used primarily as grass and annual broadleaf
herbicides; all are sorbed to soil particles, particularly to organic or
humic substances, have low water solubilities, and are practically
immobile in soils (Weber, 1990). Soil residue carryover problems
have occurred for some crops following tobacco treated with
dinitroanilines for sucker control. Rawls (1986) found that wheat
planted in the fall following flumetralin-treated tobacco showed
negative growth responses to increasing flumetralin rates at each of
four

 Page 123
locations with soil organic matter levels ranging between 0.4 and
1.2%. Bioassay studies indicated that soil concentrations of 0.05 ppm
and 0.1 ppm flumetralin reduced wheat root and shoot growth,
respectively; concentrations above 0.85 ppm retarded emergence of
wheat but not germination. Bregger (1985) showed that 1.3 kg ai/ha of
flumetralin applied to the soil surface caused stunting of corn planted
285 days later. In two experiments conducted on soils with similar
organic matter contents but with 12 or 28% clay, Sheets, et al. (1994a)
obtained slight stand and vigor reductions of wheat planted 100 days
after application of 1.34 kg ai/ha of flumetralin to tobacco at the low
clay site. However, butralin applied at 3.36 kg ai/ha did not affect
wheat stand or vigor at either location, but provided less sucker
control than flumetralin at the low clay site when both were applied
alone at the rates indicated above. Soil residue effects on following
crops may be enhanced when a dinitroaniline is used for both weed
and sucker control. Shelby, et al. (1990) found that stunting of wheat
planted in the fall following flumetralin-treated burley tobacco was
greater when pendimethalin was used as the tobacco herbicide,
whereas stunting of the tobacco planted the following spring was due
primarily to flumetralin carryover. These effects were more
pronounced when the tobacco plants received 86 mg ai/pl (1.6 kg
ai/ha) of flumetralin for sucker control rather than 29 mg ai/pl (0.5 kg
ai/ha). However, cured leaf yields appeared to be affected more by
degree of sucker control than by the stunting caused by flumetralin.
Flumetralin was approved for use on flue-cured tobacco in the USA in
1983 and butralin was approved for use in the 1997 season. While on-
farm observations with butralin are less numerous, flumetralin was
initially labeled for 1.34 kg ai/ha and substantial soil residue carryover
problems were observed by the author and others on soils with <0.5%
organic matter. In addition, over-top spray and stalk rundown
applications at the labeled rate (90 mg ai in 30 ml/pl) sometimes
restricted growth of succulent, immature upper leaves. Based on these
experiences, some due to over-application, and the work of Jones and
Rideout (1986) and Seltmann (personal communication), indicating
that lower rates of flumetralin can provide adequate sucker control,
the most commonly used rate is now 0.67 kg ai/ha (45 mg ai/pl) and
carryover complaints from growers are very rare. By comparison,
butralin is less active on suckers than flumetralin and the most
common use rate is expected to be 2.52 kg ai/ha. Based on the work of
Sheets, et al. (1994b), this rate of butralin is not expected to cause
widespread soil residue carryover problems in southeastern USA
tobacco soils.
Dinitroanilines appear to leave relatively low levels of residues in
cured leaves of flue-cured tobacco. In two experiments, Sheets, et al.
(1994b) reported that residues were usually highest for leaves of the
first priming and the highest levels found for lower leaves were 0.37
and 0.33 ppm flumetralin and butralin, respectively. Generally,
butralin disappeared more rapidly from leaves than flumetralin. In an
extensive residue sampling program conducted over the flue-cured
marketing areas in the USA during the 1993 through 1995 seasons
(Sheets, et al., 1994a; Leidy, et al., 1995, 1996), flumetralin residues
averaged 0.5 ppm over stalk positions and were not consistently
different among stalk positions. By comparison, flumetralin residues
for burley tobacco sampled in the same years averaged 1.68 ppm over
stalk positions and generally increased from the lower to upper stalk
positions. The reasons for the apparently higher residues on burley
than flue-cured are not clear, but may be because burley is harvested
sooner after treatment and high temperatures are not used during
curing. (Butralin was not included in the sampling program because it
was not labeled at that time for use in the USA.)
Systemics
Maleic hydrazide (MH) is the only true systemic and has been the
most controversial chemical used for tobacco sucker control since it
was first introduced in the USA in the 1950s. MH was the first
commercial chemical to give effective sucker control when sprayed
over tobacco with hand or tractor-mounted sprayers. It eliminated
much of the individual plant labor needed for hand suckering or
application of the oil emulsions used at that time. Consequently, MH
became widely used on both flue-cured and burley tobacco by the
mid-1960s. However, several cigarette manufacturers reported that
MH was causing undesirable changes in cured leaves compared to
hand-suckering (Coulson, 1959; Moseley, 1959). Increases in
reducing sugars and equilibrium moisture content and reductions in
nicotine and ash contents and filling value were attributed to MH use
(Chaplin, 1967; Peedin, 1970), and inconsistent preferences by expert
smoke panels for hand-suckered over MH-treated tobacco were
reported (Steffens, 1979). However, work by Peedin (1970) indicated
that some of the undesirable changes associated with MH use were
partially related to improved sucker control ratherthan to specific
physiological effects of MH on tobacco. Steffens (1979)suggested

 Page 124
that simply reducing the number of meristemic regions with chemical
sucker control could have profound effects on the physiology of the
plant. The data of Seltmann (1978), however, indicated that the higher
reducing sugar and equilibrium moisture contents associated with
MH-treated tobaccos could not be explained by better sucker control
alone.
The international trade of MH-treated tobacco is affected by MH
residues which are the highest of any pesticide used on tobacco
(Collins & Hawks, 1993). Some major flue-cured countries such as
Brazil, Canada and Zimbabwe prohibit MH use because several
countries in the European Union have a residue tolerance of 80 ppm
on manufactured products. However, Meyer, et al. (1987) reviewed
factors affecting MH residues and their toxicological implications and
cited no convincing evidence that the potassium salt formulation used
worldwide is harmful to smokers. Further, the US Environmental
Protection Agency reviewed data from numerous toxicological studies
with MH in the early 1980s and did not establish MH residue
tolerances for tobacco products manufactured in the USA.
Maleic hydrazide absorption is most efficient under conditions of
good plant turgor and high humidity (Meyer, et al., 1987) and its use
is not recommended in hot, dry conditions. After absorption, it is
rapidly translocated in both the xylem and phloem to meristems where
it inhibits cell division but not cell elongation. Maleic hydrazide is
relatively stable within the plant and exists in free and bound forms.
Free MH includes the water soluble portion in the leaf and the
residues on the leaf surface. Bound MH is thought to be associated
with cell walls and glucosides and may not participate in
physiological responses attributed to MH. Glucoside formation may
explain why Seltmann and Peedin (1972) observed poorer sucker
control when MH was applied in the evening rather than in the
morning or mid-afternoon; plant carbohydrate levels are higher in the
early evening due to accumulation during the day (Meyer, et al.,
1987).
In addition to its metabolic stability, the vapor pressure of MH is
essentially zero and MH is not significantly degraded by UV radiation
or by the highest temperatures used during flue-curing (Collins &
Hawks, 1993). Therefore, treated leaves almost always contain MH
residues (Meyer, et al., 1987) which decline very little during curing
(Sheets & Seltmann, 1985) and storage for 24 months (Steffens,
1979). Maleic hydrazide residues are directly related to rate of MH
application (Hunt, et al., 1977; Sheets & Seltmann, 1985; Cui, et al.,
1995), but MH is very water soluble and published residue levels for
cured leaves vary widely, probably as a result of varying rainfall
and/or heavy dew events within several days after application. Cui, et
al. (1995) reported that MH residues on green, upper leaves of burley
tobacco declined about 50% during the first 8 days after MH
application at the labeled rate and then remained relatively stable until
harvest 16 days later. Possible reasons for the initial decline were not
stated but the rate of decline best fitted (r = 0.99** which is highly
significant) the power model:
MH (ppm) = 502 × days-0.84
where days = days after MH application. Hunt, et al. (1977) found
that cured leaves from the lower stalk positions generally contained
higher residues than those from the middle and upper positions, but
residues decreased with time after application, leading to the
recommendation of allowing a week between MH application and
harvest of lower leaves. A 5-year study conducted over 29 locations in
North Carolina indicated that when the labeled rate of MH (2.5 kg
ai/ha) was used only once and leaves were harvested 7 or more days
after application, residues for composited, whole-leaf samples varied
widely over locations and years but averaged 73 ppm for the 163
samples analyzed. The range in residue levels varied from
nondetectable to a high of 351 ppm and exceeded 80 ppm in 36% of
the samples (Sheets & Nelson, 1989). These authors found a low but
significant negative correlation between average MH residue level and
yield/ha for some locations and years, but the relationship was not
strong enough to use yield as a predictor of MH residues.
Because of its high water solubility, residues of MH on tobacco can be
substantially and consistently reduced by relatively small and properly
timed overtop applications of water after MH application. Sheets and
Seltmann (1982) reduced degree of sucker control as well as MH
residues by initiation of 2 to 3 cm of sprinkler irrigation 6 hours after
MH application; irrigation applied 12 hours after MH application also
reduced residues but did not consistently reduce sucker control.
Irrigation applied 24 or 48 hours after MH application did not affect
residues or sucker control compared to the nonirrigated control. These
results were generally confirmed in a later study (Seltmann & Sheets,
1987), and the authors found that reapplication of a half rate of MH
following rain occurring between 3 and 12 hours after the initial MH
application maintained sucker control without increasing residues
above 80 ppm. Jennette, et al.

 Page 125
(1995) applied simulated rainfall ranging from 0.013 to 1.27 cm at
four locations over 2 years and found that as little as 0.5 cm of water
applied 24 hours after MH application reduced residues more than
50% on both upper and lower green leaves compared to the
nonirrigated control. The residue reduction occurred in a linear or
two-part segmented fashion as simulated rainfall increased from none
to the highest level, and the effect was most consistent on upper
leaves which generally had two to three times higher residues than
lower leaves. In cases when the reduction was segmented, the 'join
point' between linear segments often occurred between 0.13 and 0.25
cm of irrigation. Degree of sucker control was not reduced by any
level of irrigation. Collectively, these studies indicate that MH
residues can be effectively reduced by overhead irrigation applied
between 12 and 24 hours after MH application without reducing
degree of sucker control. The amount of water to apply is more
speculative than the time of water application, but maximum residue
reduction should be possible with 1 to 1.5 cm of water.
The method of MH application may affect residue levels on cured
leaves. Jennette, et al. (1995) reported that 2.52 kg ai/ha of MH
applied as a fine spray with low volume (296 L/ha) and high pressure
(276 kPa) resulted in higher residues than the same rate of MH
applied as a coarse spray with high volume (468 L/ha) and low
pressure (138 kPa). On average, the coarse spray reduced residues by
5% on lower green leaves and 34% on upper green leaves, resulting in
a 23% reduction for composited cured leaves. These authors
speculated that fine droplets were more mobile and therefore more
likely to attach to the undersides of leaves where wash-off is less
likely. In a similar study, however, Yelverton, et al. (1995) did not
obtain differences for residues or sucker control between coarse and
fine sprays of the same MH rate when the delivery volume was the
same (470 L/ha) for both application methods but pressures were 150
and 300 kPa for the coarse and fine sprays, respectively. Cui, et al.
(1995) applied the same total rate of MH to burley tobacco in single
or split applications in 392 L/ha per application or as a single
application in 196 L/ha. Compared to the single application of MH
with high water volume, cured leaves treated with the split application
had intermediate MH residues and those treated with the low spray
volume had the highest residues, particularly those from the middle
and upper node positions. They speculated that the higher residues
with the split application occurred because the second application was
made seven days closer to harvest, while the highest residues from the
single low water application may have been the result of better MH
absorption because of its higher concentration in solution. If so, the
apparent positive relationship between MH concentration and
absorption may help explain why Jennette, et al. (1995) obtained
higher residues with fine, low volume sprays than with high volume,
coarse sprays, while Yelverton, et al. (1995), who used the same
volume for both coarse and fine sprays, did not. In a 3-year study,
McKee (1995) also obtained higher residues on Maryland tobacco
when the same rate of total MH was applied in two applications rather
than one, using the same delivery volume and pressure for both
treatments, but he did not speculate on why the difference occurred.
Maleic hydrazide is now used on most of the fluecured tobacco grown
in the USA. The low availability and high cost of labor, coupled with
recent legislations mandating federal salary and social security taxes
and certain housing and worker protection requirements for migrant
labor, are encouraging tobacco growers to replace manual labor with
machinery. Therefore, hand application of contacts and/or contact-
local systemic is declining and most sucker control chemicals are now
applied with tractor-mounted or high clearance power sprayers. Past
experience and research indicated that acceptable sucker control could
not be achieved consistently with contacts and/or contact-local
systemics applied in single or multiple applications with power
sprayers, especially for mechanically harvested tobacco which would
include suckers as foreign matter. Therefore, the preferred sucker
control program from the mid-1970s until recent years was two fatty
alcohol contact applications followed by MH. Under normal field
conditions, MH provides adequate sucker control for 6 to 7 weeks
after application (Collins & Hawks, 1993). As production units
became larger and more mechanized and farmers adopted improved
cultural practices and varieties which increased yields but extended
the harvest season, farmers quickly discovered that the labeled rate of
MH would no longer provide season-long control of suckers.
Therefore, excessive rates of MH were often used, sometimes as
second applications during the harvest season to control late season
sucker regrowth. Although flumetralin was approved for use in the
USA in 1983, it was not widely accepted by growers initially because
of its relatively high cost and potential for soil residue carryover when
used at the labeled rate of 1.34 kg ai/ha. Therefore, during the late
1980s and early 1990s, MH residues in US fluecured tobacco
averaged 60 to 80% higher than the 80 ppm tolerance level previously
established by the Federal Republic of Germany

 Page 126
(Leidy, et al., 1997) and then being considered by several other
countries in the European Union, the largest importer of US tobacco.
A reduced rate of flumetralin in a tankmix combination with MH was
evaluated in the early 1980s (Ben Whitty, personal communication,
University of Florida). With this approach, the growth of suckers
missed by flumetralin is controlled by MH. In further testing by the
Regional Tobacco Growth Regulator Committee, 0.67 kg ai/ha of
flumetralin tankmixed with the labeled rate of MH and applied after
two contact fatty alcohol applications provided better and longer
lasting sucker control than the alcohols followed by MH alone. The
tankmix was labeled in 1991 and is now the most consistently
effective and widely used treatment (Table 5.9) for mechanical sucker
control on flue-cured tobacco in the USA. Flumetralin is also effective
when applied alone 2 to 3 weeks after MH to reduce lateseason sucker
regrowth, but application time is critical and it must be applied before
the sucker buds reach 2.5 to 3 cm in length to be as effective as shown
in Table 5.9. The use of flumetralin, primarily in a tankmix with MH,
has been a significant factor in reducing MH residues in US flue-
cured tobacco because it has allowed growers to obtain season-long
sucker control without using excessive rates of MH. Based on samples
collected by the Flue-Cured Stabilization Corporation, average MH
residues declined from 147 ppm in 1990 to 85 ppm in 1996 (Peedin &
Priest, 1996). Use of butralin at the 2.52 kg ai/ha rate in a tankmix
with MH or 2 to 3 weeks after MH alone should give results
comparable to those in Table 5.9, but data obtained in North Carolina
during the 199596 seasons indicated
Table 5.9 Sucker numbers and weights
allowed by several chemical sucker control
treatments applied with power sprayers in
North Carolina, 199194 (G.F. Peedin,
unpublished data).
 Suckers per Weight per
Treatment1 plant sucker

(No) (g) (g)

MH alone 2.74 128 47
MH and 0.26 29 112
flumetralin,
tankmix
0.09 13 144
1Each treatment was preceded by two fatty
alcohol applications of 4% and 5%,
respectively; rates of MH and flumetralin
were 2.52 and 0.67 kg ai/ha, respectively;
spray volume was 468 L/ha for all
applications. All results are the average of 25
on-farm tests.

that 2.52 kg/ha of butralin is slightly less effective than 0.67 ks/ha of
flumetralin in the MH tankmix (Peedin, unpublished data). However,
meaningful differences between the two products would be expected
only when the harvest season is abnormally extended by drought
and/or excessive N application. When N rate is excessive, sucker
growth is stimulated and ripening is delayed, which increases the
probability of sucker regrowth in extended harvest seasons (Yelverton,
1994). Under these conditions, suckers are difficult to control with
any combination of suckercides.
Several tobacco breeders have explored the possibility of breeding
tobacco varieties with fewer and/or smaller suckers. Seltmann and
Sisson (1993) evaluated several breeding lines with reduced sucker
growth against two commercial flue-cured varieties to determine
whether acceptable sucker control could be achieved on the low-
sucker genotypes with reduced rates of MH. All plants except the
hand-suckered controls were treated with two fatty alcohol contact
applications followed by the full or half rate of MH. Chemical control
was more effective on the low-sucker genotypes than on the varieties.
Two genotypes with the lowest sucker growth regardless of the
chemical treatment produced significantly lower yields than the
varieties. However, one genotype with intermediate sucker growth
produced yields equal to those of the varieties and was selected for
further evaluation. These workers concluded that development of
varieties with lower sucker number and size is achievable and should
make it possible to obtain sucker control with reduced rates of
suckercides and, therefore, reduce chemical residues on tobacco.
Harvesting and Ripeness
a
Number of Primings
Flue-cured, Oriental and cigar wrapper are the only tobacco types
whose leaves are harvested (primed) in sequence as they ripen from
the bottom to the top of the plant. In most flue-cured countries, several
ripe leaves per plant are removed by hand in each of several passes
over the field during a period of 6 to 12 weeks after topping,
depending on temperature, rainfall/irrigation, soil productivity and N
fertilization. The number of primings varies within and among
countries, depending on labor availability and cost, level of
mechanization and buyer preferences. In Zimbabwe, where
production units are relatively large but labor is plentiful and
inexpensive and 98% of the crop is

 Page 127
exported (Zimbabwe Tobacco Industry Council, 1990), about two
leaves per plant are removed in each of 10 primings. In Brazil, where
production units average only 2 to 3 ha and 90% of the harvest labor
is provided by family members and neighbors (Association of
Brazilian Tobacco Growers, 1994), five to seven primings are made
on plants topped at 18 to 20 leaves. In the USA, almost 50% of the
crop is primed mechanically and the majority of the land area is
harvested in three primings, compared to five primings 15 years ago.
Unfortunately, as size of production units has increased in the USA, a
substantial portion of the land area is also harvested in two primings
because the predominant variety (K 326) has good holding ability in
the field, labor costs and machinery use are reduced, movement of
harvesting equipment among farms is less and ultimately because
buyer discrimination against tobaccos mixed over stalk positions has
been minimal to date.
Most published research on reducing number of primings has been
conducted in the USA to evaluate possible impacts of mechanized
harvesting on the agronomic and chemical properties of flue-cured
tobacco; the results have been inconsistent. Gooden, et al. (1976a,
1976b) found that harvesting 18 leaves per plant in six or three
primings had very little effect on yield, visual quality or
concentrations of total N, alkaloids and reducing sugars of cured
leaves. However, in 21 on-farm tests conducted over 3 years in the
late 1960s, Collins and Hawks (1993) reported a 5% yield reduction
with three primings compared to the normal five to seven primings
routinely used in North Carolina at that time. Compared to four
primings, Chaplin (1975) obtained 15 and 20% yield reductions when
normally topped tobaccos were harvested in two or one primings,
respectively. Average market price and total alkaloids of cured leaves
were not affected by priming number, but reducing sugars were higher
in tobaccos harvested only once or twice. Jones (1988) evaluated five,
three and two primings in seven on-farm tests over 2 years in Virginia
and obtained 13% reductions in both yield and value per hectare with
three or two primings; grade index was not consistently affected by
number of primings. In another study involving time of harvesting
(Johnson, 1966), tobacco harvested in three or two equal primings (by
leaf number) produced yields similar to that harvested in six primings,
but the earlier harvest needed for optimum yield with three or two
primings generally reduced average market price compared to the six-
priming control, apparently the result of harvesting under-ripe
tobacco. Generally, time of harvest had a greater influence on
chemical and physical properties of cured leaves than did number of
primings.
The twopriming system popular among farmers in the southeastern
USA involves removing five to seven leaves at the first priming and
the remaining leaves 4 to 6 weeks later, often following treatment
with ethephon. Suggs, et al. (1989) found that harvesting the bottom
four to six leaves at optimum ripeness, followed by harvest of all
remaining leaves 2 or 4 weeks later, did not reduce yield or value per
hectare compared to a four-priming control. However, the lowest
average market prices were associated with harvesting the remaining
leaves 2 (under-ripe) or 6 (over-ripe) weeks after the lower leaves
were harvested. The agronomic results obtained by Peele (1994)
generally agree with those of Suggs, et al. (1989). However,
compared to the four-priming control, the twopriming system resulted
in slightly lower yields in each of the three experiments and reduced
grade index (but not average market price) in two of three
experiments. Chemical compositions of cured leaves were influenced
more by maturity at harvest than by number of primings. In an earlier
study, Peedin (unpublished data) conducted nine experiments over 3
years and obtained a 4% yield reduction with the twopriming system
compared to a four-priming control. There was no measurable effect
on value per hectare because both grade index and average market
price were increased with the twopriming system, which resulted in a
higher proportion of tobacco graded in the more valuable leaf (B)
category. However, compared to the four-priming control, harvesting
all leaves in a single priming when the upper leaves were judged to be
mature reduced yield and value per hectare by 13% and 10%,
respectively, but increased grade index by 8%. In a later study
conducted in South Carolina, Gooden and Rideout (1997) obtained
results very similar to those of Peedin (unpublished) for each of the
three varieties tested and concluded that flue-cured tobacco should be
harvested three or four times to offer purchasers the opportunity to
select tobaccos from the various stalk positions.
Because of the development of mechanical harvesters in the USA with
the capacity to remove 15 to 17 leaves per plant in a single pass, once-
over harvesting of plants topped low enough to accommodate the
harvesting capacity of the machines has been studied. Compared to
four primings on normally topped plants, Chaplin (1975) reduced
yield by an average of 38% and increased total alkaloids and reducing
sugars by over 20% by topping plants at 61 cm and harvesting only
once. Delaying harvest of low-topped, once-harvested plants by 1 and
2 weeks progressively reduced yield but

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increased visual quality as measured by average market price.
Working with 12-and 15-leaf topping heights, Miner (1980a; 1980b)
found that multipass and onceover harvests resulted in similar grade
indices but yield was decreased by 8% with onceover harvesting;
differences in chemical composition of cured leaves were not
consistent and were related to the harvest intervals selected between
the harvest methods. Gooden, et al. (1976b) compared onceover
harvesting with a range of multiple harvests at a constant leaf
population and concluded that onceover harvesting resulted in a
mixing of cured leaves varying widely in chemical and physical
properties, but generally total alkaoids and N were increased and
reducing sugars were decreased by fewer primings.
Collectively, the studies above indicate that harvesting flue-cured
tobacco in less than four primings in the southeastern USA would be
expected to reduce yield because fewer leaves are harvested at
optimum maturity; the greatest yield reduction and mixing of stalk
positions would occur with only one priming. The two-priming
system discussed above minimizes yield losses because lower leaves
are not allowed to deteriorate before the first harvest and upper leaves
are usually allowed to mature before the last harvest is made.
However, if the variety has poor holding ability and/or harvesting is
delayed by unfavorable weather conditions or limited curing facilities,
the yield reduction is likely to be greater. Because grading systems are
subjective, the mixing (i.e. hiding) of less valuable lower stalk
tobaccos with more valuable upper stalk tobaccos, which occurs in
varying degrees with all reduced priming systems, may actually
increase measures of visual quality, thereby offsetting some of the
value of the yield loss. Further, reduced priming systems appear to
have minimal and/or inconsistent effects on the 'average' chemical and
physical properties of cured leaves, which generally are affected more
by time of primings rather than number of primings. Most
importantly, distinct chemical and physical properties traditionally
have been provided to the purchaser in distinct and separate stalk
positions, but with reduced primings, leaves remain hopelessly
intermixed by both stalk positions and ripenesses (Weybrew, 1983),
which severely limits the cigarette manufacturer's flexibility to make
specific blends preferred by customers worldwide. Therefore, the
grower's decision on number of primings should be based more on the
industry's ability to consistently provide reasonably priced, quality
products to customers than on perceived economic advantages which
may ultimately reduce demand for his raw material.
b
Ripeness at Harvest
Based on some of the priming studies above, it is obvious that degree
of leaf ripeness at harvest is an important factor determining the
suitability of cured leaves for use in smoking products. Ripe tobaccos
cure more easily, respond more readily to aging and deliver a more
flavorful and palatable smoke, while immature tobaccos have long
been associated with poor aroma and smoke taste (Moseley, et al.,
1963). Both Weybrew, et al. (1984) and Moseley, et al. (1963) gave
similar descriptions of leaf ripeness in the field. As green leaves ripen,
they droop away from the stalk to assume a more horizontal attitude,
lose their sheen and appear more velvety, and the green color fades as
chlorophyll degrades and a yellowish color emerges from the apical
margins towards the bases of the leaves. The basal halves of the
midribs also change from light green to milky white. These color
changes are usually the principal guides in judging ripeness. However,
as leaves ripen, they develop 'grain' or mounds between the small
veins, become more turgid and do not wilt readily, and snap crisply
and cleanly from the stalk. With under-ripe leaves, flaps of stalk
epidermis usually tear away with the petioles. Weybrew, et al. (1984)
stated that physiological maturity and ripeness are not the same;
maturity is the attainment of maximum leaf dry weight which
indicates the transition from growth to senescence, while ripeness
occurs about 12 days after maturity. However, rate of ripeness is
affected by many cultural and environmental factors and may be
reduced by use of today's improved cultural practices and cultivars
which encourage high yields.
The effects of ripeness on the agronomic, chemical and physical
properties of flue-cured tobacco are somewhat variable, probably
because of variable climatic conditions within and among the studies
reported below. Compared to harvesting leaves judged to be ripe,
harvesting about 1 week under-ripe may not affect yield (Moseley, et
at., 1963) or increase yield (Weybrew, et al., 1984), but delaying
harvest to the overripe stage generally decreases yield (Moseley, et
al., 1963; Weybrew, et al., 1984; Tucker, 1987; Flower, 1995). The
yield reductions associated with overripeness are the result of
deterioration of the senescencing leaves and net respiratory losses of
carbohydrates.
The highest visual quality ratings are normally associated with ripe
tobaccos. In three experiments, Weybrew, et al. (1984) reported
consistently higher grade indices for ripe tobacco, and overripe
tobaccos had higher grade indices than under-ripe tobaccos in

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two of three experiments. Moseley, et al. (1963) did not report visual
quality ratings but observed that better leaf color and texture were
associated with ripeness, while under-ripe cured leaves were dull,
slick and soggy. Hawks, et al. (1976) found that harvesting as fast as
the leaves could be cured resulted in lower grade indices than
harvesting as slow as possible without leaf deterioration, but harvest
rate did not affect yield. Tucker (1987) found that normal N
fertilization and ripe harvesting generally produced higher grade
indices and average market prices than combinations of low or high N
fertilization and harvesting under-ripe or over-ripe. In one experiment
with an unusually wet early season, however, both grade index and
average market price of over-ripe tobaccos were less than those of
under-ripe tobaccos, regardless of N rate.
Cured leaf colorgrade distributions indicated that:
(1) tobaccos harvested ripe produced the highest weight percentage of
lemon (L) and orange (F) grades,
(2) the percentage of variegated (K) tobaccos increased with both
higher N fertilization and greater ripeness, and
(3) the percentage of green (G) and variegated mixed (KM) tobaccos
decreased with greater ripeness (Tucker, 1987).
The effects of ripeness on the percentages of K and G tobaccos were
similar to those reported by Weybrew, et al. (1984). However, in six
Zimbabwean experiments summarized by Flower (1995), grade index
and colorgrade distribution were not consistently affected by ripeness
at harvest, and results were confounded by environment and curing
regimes. Flower (1995) suggested that further progress will be made
in ripeness experimentation only if ripeness is measured less
subjectively and the effects of climate and curing are evaluated.
In smoke preference taste tests, Moseley, et al. (1963) reported that
ripe tobaccos were preferred over under-ripe tobaccos, but that both
quantity and quality of aroma were less for over-ripe tobaccos.
Weybrew, et al. (1984) obtained unexpected results; smoke panels
from five domestic cigarette manufacturers judged cigarettes made
from under-ripe tobaccos yellowed longer than normal during curing
to be milder and preferable to over-ripe tobaccos yellowed for shorter
periods of time. In most instances, the panelists selected the cigarettes
with the lowest nicotine contents, which averaged 3.28% in the test
cigarettes. However, tobacco company leaf experts' visual appraisal
favored tobaccos harvested ripe with normal to long yellowing
periods. Peele (1994) reported that immaturity at harvest had the most
consistent adverse effect on smoke acceptability of twice-primed
tobaccos compared to those of the four-primed control, and when
twice-primed tobaccos were allowed to ripen their smoking
characteristics were similar to those harvested in four primings.
Delayed harvesting affects the physical and chemical properties of
cured leaves, but the reported effects are sometimes inconsistent.
Moseley, et al. (1963) reported that increasing ripeness decreased
moistureholding capacity, reducing sugars, the sugar/nicotine ratio
and total and protein N. However, filling capacity, strip yield,
nicotine, the nicotine/total N ratio and petroleum ether extracts tended
to increase with increasing ripeness. Weybrew, et al. (1984) found that
increasing ripeness generally increased nicotine, starch, reducing
sugars and strip yield but had no consistent effect on equilibrium
moisture, filling capacity, lamina density, the sugar/nicotine ratio and
level of aroma. Concentrations of most nitrogenous constituents,
including total N, decreased with delayed harvest. Generally, leaf
chemistries were influenced more by stalk position than by either
ripeness at harvest or curing schedule.
Curing and Marketing
Wood smoke inside the curing barn was used initially to reduce rots
and molds and to prepare the leaf for storage and transport to the
market. Growers in Virginia and North Carolina began to use heat to
cure tobacco to a lighter color in the early nineteenth century. The use
of flues to transfer heat from a firebox was patented by Dr Davis G.
Tuck of Halifax, Virginia in 1831 (Peele, et al., 1995), although flue-
curing was not completely adopted in the USA until almost 100 years
later. As mentioned previously, the 'color fixing' step of flue-curing
was accidentally discovered by the 18-year old slave, Stephen Slade,
in 1839. Soon thereafter, cured leaves with bright orange-lemon color
became associated with tobacco grown on relatively infertile soils and
cured with open charcoal fires. Around 1870, R.L. Ragland of
Virginia proposed four sequential stages of the flue-curing process:
yellowing, color-fixing, leaf drying and stem (midrib) drying. While
curing facilities and technology, varieties and cultural practices have
changed drastically since that time, use of Ragland's proposed curing
stages remains essentially unchanged for successful curing today.
However, in recent publications, some workers con-

 Page 130
sider color-fixing as the first step in leaf drying (Johnson, 1974; Peele,
et al., 1995).
a
Curing Systems and Energy Sources
Of the seven major tobacco types produced in the world, fluecured is
the only type that requires substantial amounts of energy for curing. In
1993, approximately 82% of the world's fluecured tobacco was cured
in conventional barns constructed of wood, burnt bricks, cement
blocks or wood poles and mud, depending on local availability and
cost of construction materials (Campbell, 1995b). With conventional
systems, groups of two to four harvested leaves are placed on wooden
or metal 'sticks' which are loaded into and removed from the barn by
hand; the maximum cured weight per stick seldom exceeds 1 kg.
The remaining 18ø/o of the flue tobacco in 1993 was cured in tightly-
constructed and better insulated metal or wood bulk barns that
allowed more precise control of temperature, humidity and ventilation
within the barn. These advantages of bulk curing, plus the denser
packing of green tobacco made possible with forced air circulation,
reduced energy consumption per kg of cured tobacco compared to
conventional barns (Peele, et al., 1995). More recent improvements in
bulk curing include automatic temperature controls that reduce human
error and require less management time during curing (Glover, 1989).
Bulk curing systems reduce labor requirements because individual
leaves are handled less during the loading and unloading of curing
containers, particularly with the box system. However, the bulk rack
system is most widely used in the world and each rack is filled by
hand and yields 8 to 12 kg of cured tobacco, depending on leaf size
and stalk position. The box system is widely used in the USA and the
boxes may be filled manually after harvesting or mechanically as the
leaves are primed with automatic harvesters; each box yields 150 to
180 kg cured tobacco.
Two-thirds of the world's curing energy for fluecured tobacco is
obtained from coal, notably in China, Zimbabwe, South Africa, and
India (Campbell, 1995b). The remaining energy is obtained about
equally from wood and petroleum products (various fuel oils and
natural or liquid gases). Wood is the predominant energy source in
South America and Asia, while oil and gas are the major sources in
North America and some European countries where bulk curing is
widely practised. Oil and gas are also used in conventional barns in
Mexico, Argentina, the Dominican Republic and Indonesia.
Deforestation is a problem in many tobacco-producing countries,
although one detailed study in 69 countries (excluding China)
representing 66% of the flue-and fire-cured tobacco produced in 1986
indicated that tobacco accounted for only 1% of the total wood
consumption in those countries (Campbell, 1995b). The tobacco
industry has encouraged tree growing by tobacco farmers in wood
deficit and prospective deficit countries for years. Campbell (1995b)
cited successful tree growing programs in Brazil and Kenya. In Brazil,
the Brazilian Tobacco Growers' Association promoted and financed a
program to make every tobacco farm self-sufficient in reforested
wood without resorting to native forests. Approximately 1 ha of trees,
primarily ratooned Eucalyptus species, replanted about every 28
years, will sustain the average Brazilian fluecured farm of 2 ha. In
Kenya, farmers growing flue-, fire-cured or burley tobacco are
required to plant and grow their own tree seedlings before entering
into a contract with a tobacco company. The farmer's seedlings must
also be available for planting by the general public. The Kenyan
tobacco industry is currently self-sufficient in wood fuel.
Very useful curing research in countries such as Zimbabwe, Brazil and
India has emphasized energy efficiency by improving temperature and
humidity control in barns, using forced air flow in conventional barns
and furnaces, improving heat exchangers and redesigning furnaces
and curing barns. The average Brazilian farmer uses about 4 kg wood
per kg of cured tobacco, considerably less than the world average for
wood consumption based on data cited by Campbell (1995b).
Improved curing systems in Zimbabwe use 1.0 to 1.6 kg coal per kg
of cured tobacco, compared to about 2.5 kg for conventional barns
(Andrew Brooker, personal communication, formerly Tobacco
Research Board, Harare, Zimbabwe). Most improved systems have
forced air furnaces. In India, where about 60% of the tobacco is cured
with wood and the remainder with coal, average fuel consumption per
kg cured tobacco is about 4.5 or 3.0 kg for wood or coal, respectively
(Srinivas Putchala, personal communication, ITC Limited,
Rajahmundry, India). Watkins (1990) measured fuel oil and liquid
propane (LP) gas consumption for fluecuring in a large number of on-
farm tests, primarily with rack-type bulk barns, and reported
consumptions of 0.9 and 1.0 L/kg cured tobacco, respectively. He also
suggested the following practices to maximize energy efficiency in
bulk barns:
(1) Stop air leaks in the barn structure, particularly around doors and
between the barn walls and the cement pad.

 Page 131
(2) Use a hygrometer to determine when and how much to ventilate.
(3) Inspect and maintain burners routinely, especially before the start
of each curing season.
(4) Install insulation if necessary, but use only materials and
installation methods approved by tobacco purchasers.
(5) Harvest only ripe tobacco to reduce curing time.
(6) Load racks or boxes uniformly to assure even and quicker drying.
(7) Assure air seals between racks or boxes to force air through the
tobacco rather than between curing containers.
b
The FlueCuring Process
Fluecuring requires 5 to 7 days for completion and removes about
97% of the moisture from the harvested leaf (Peele, et al., 1995).
However, fluecuring is a biological as well as a drying process. The
major objective is to maintain the potential quality of freshly
harvested ripe leaves by providing a scheduled regime of temperature
and humidity control within the barn. This optimizes and preserves
the products of certain desirable chemical and biological changes
needed in cured tobacco suitable for the manufacture of cigarette
products that are acceptable to the consumer (Collins, 1983). The
curing schedule varies with the barns used, the stalk position from
which the leaves were taken, the weather conditions during harvesting
and curing and the ripeness and moisture content of the leaves at
harvest (Hawks, 1975). Johnson (1974) stated that curing is very
much an art since curing conditions must be modified according to the
variability of the leaves at harvest. The major process variables under
the curer's control are air temperature, relative humidity, air velocity
(in forced air systems), and the time variation of these variables. Rate
and extent of biochemical reactions are controlled by leaf temperature
and moisture content, while air velocity provides uniformity of curing
conditions and facilitates the timely removal of moisture from the
curing system. Consequently, considerable and varied experience as
well as a thorough knowledge of curing principles are needed to
produce consistently successful cures.
The chemical and biochemical changes that occur during fluecuring
and the effects of altered curing regimes on leaf chemistry and quality
have been reviewed by several authors (Johnson, 1974; Weybrew, et
al., 1984; Peele, et al., 1995; Jones & Wilkinson, 1996). Chemical
transformations during curing are essentially extensions of those
initiated during senescence in the field (Long & Weybrew, 1981). The
tobacco leaf is metabolically active at the beginning of curing and
biochemical processes continue until terminated by high temperature
and/or desiccation (Johnson, 1974). The yellowing stage is the period
of major biochemical conversion which continues into the leaf drying
stage. Peele, et al. (1995) summarized the most important changes as
follows:
(1) chlorophyll degradation which unmasks the yellow carotenoid
pigments,
(2) hydrolysis of starch to free sugars and partial respiration of these
sugars to carbon dioxide,
(3) hydrolysis of proteins into free amino acids and the subsequent
reaction of free amino acids with free sugars tO form Amadori and
Maillard compounds important to flavor and aroma; these are
primarily high temperature products formed during the leaf and stem
drying stages,
(4) variable changes in polyphenols which usually have no
undesirable effects on fluecured quality unless the leaves are over-
yellowed to cause oxidative browning,
(5) thermal degradation of leaf surface diterpenes and sugar esters into
more volatile compounds, and
(6) the conversion of nitrate to nitrite and its subsequent reaction with
alkaloids to form tobacco-specific nitrosamines.
Many of these changes result in the production of flavor and aroma
compounds characteristic of fluecured tobacco (Peele, et al., 1995).
It is widely accepted that the potential quality of tobacco is
determined in the field and that harvested leaves with poor quality
characteristics are not improved during curing, even when curing
procedures are ideal (Walker & Stier, 1987). Improper curing can,
however, reduce the final quality of leaves with either good or poor
quality potential. Therefore, the first requirement for proper curing is
to start with uniformly ripe tobacco. Peele, et al. (1995) stated that no
amount of curing skill can completely compensate for the failure to
harvest tobacco at the proper stage of maturity. Although Weybrew, et
al. (1984) showed that the visual quality of under-ripe or overripe
leaves can be improved by extending or reducing the normal
yellowing period, respectively, the highest levels of chemical and
physical quality components were associated with cured leaves which
were harvested when judged to be ripe or slightly overripe. Peele, et
al. (1995) reviewed several other advantages of curing ripe compared
to under-ripe or overripe tobacco:

 Page 132
(1) ripe leaves require less curing time and color more uniformly,
while leaves with variable ripenesses may result in overcuring of ripe
leaves and undercuring of under-ripe leaves,
(2) tobacco harvested under-ripe often has a greenish cast after curing
and traditionally a lower selling price because of undesirable aesthetic
and smoking characteristics,
(3) ripe tobacco usually has a more desirable leaf texture and color
than under-ripe tobacco, and
(4) under-ripe leaves dry more rapidly during yellowing and leaf
drying, which may explain the high frequency of green color
associated with under-ripe tobacco after curing.
The principles of flue-curing are the same for both conventional and
bulk barns, although the curer has greater and more timely control of
the processing variables in bulk barns. The dry and wet bulb
temperatures, relative humidities and their time variation for the four
major curing stages of a typical flue-curing schedule in bulk barns are
shown in Fig. 5.2. Yellowing is considered as the most important
stage of curing because the potential quality of leaves is either
maintained or lowered during this period (Hawks, 1972). For the
important biochemical conversions initiated in the field to continue
satisfactorily during curing, the leaves must maintain a relatively high
moisture content during yellowing. Therefore, yellowing normally
occurs with dry bulb temperatures of 35 to 37°C and humidities of 85
to 90% for 36 to 72 hours. During this period, the loss of chlorophyll
and moisture from the leaves should be monitored closely. When
green leaves are harvested, they have relatively high starch and low
 Fig. 5.2
Typical flue-curing schedule for normal, ripe tobacco in bulk barns (D.M. Peele,
unpublished data, R.J. Reynolds Tobacco Company).
sugar contents, both of which are undesirable in the cured product.
The starch level in harvested leaves limits the sugar content of cured
leaves, but the extent of the starch to sugar conversion is largely
determined by the curing process (Papenfus, 1996). The conversion of
starch to sugar and the color change from green to yellow, although
independent processes, occur simultaneously and at about the same
rates (Hawks, et al., 1975). Therefore, sugar accumulation is near
optimum when most of the chlorophyll has degraded, and color-fixing
should be initiated at this time.
Incomplete conversion of starch results in cured leaves with a thick,
'soapy' feel, a high propensity for 'tar' delivery and a bitter taste and
metallic-like aftertaste (Papenfus, 1996). Extended yellowing
exacerbates respiratory losses of converted sugars, and perhaps other
carbohydrates, which lowers yield as well as sugar content and the
sugar:nicotine ratio of cured leaves (Garvin, 1988). Papenfus (1996)
stated that overcolored leaves are thinner than normal and at the
middle and upper stalk positions, the thinness is exhibited along the
leaf midribs as paper-thin tissue which becomes 'trash' during
processing and contributes to lower lamina:stem ratios. Extended
yellowing may also cause 'sponging', a curing disorder related to the
oxidation of polyphenols to compounds that impart an undesirable
dull brown color to cured leaves (Garvin, 1988). Lower stalk leaves,
especially if overfertilized with N, are most susceptible, but
polyphenol oxidation can also cause dullness of middle and upper
stalk tobaccos (Papenfus, 1996). Sponged tobaccos usually have low
sugar:nicotine ratios and, therefore, receive lower market prices than
properly cured tobaccos.
The moisture content of a fresh, ripe leaf varies between 80 and 90%,
but is reduced to about 3% during curing. One of the most critical
aspects of curing is controlling the moisture content of the leaves
during yellowing (Hawks, 1972; Papenfus, 1996). Enough moisture
must be maintained in the leaf so that the desirable chemical and color
changes can occur unimpeded. Drying too quickly usually results in
greenish colored cured leaves which indicate undesirably high starch
contents. Conversely, enough drying must occur so that the leaf will
dry without darkening or scalding when the temperature is increased
during the leaf drying stage. Papenfus (1996) suggested minimum
ventilation (i.e. moisture loss) for the first 12 (lower leaves) to 24
(upper leaves) hours and then increasing ventilation so that about 30%
of the moisture is lost by the end of yellowing. For normal ripe
tobacco, this can usually be accomplished by main-

 Page 133
mining a 1°C differential between the dry and wet bulbs for the first
12 to 24 hours and increasing the differential to 2.53°C by the end of
yellowing. If successful, the yellowed leaves should be completely
wilted and some of the leaf tips dry and crisp when the yellowing
stage is complete. Johnson (1974) reported more rapid yellowing at
higher temperatures and when leaves were slightly wilted.
The rate of drying during yellowing is controlled primarily by
ventilation and, depending on the characteristics of the tobacco and
weather conditions at harvest, may vary substantially. Fresh leaves
from lower stalk positions usually have higher moisture contents than
those from upper positions. Therefore, faster drying is usually
required for lower than upper stalk tobacco and for tobacco harvested
during wet compared to dry weather. Hawks (1972) stated that ripe
lower leaves harvested in a rainy period yellow relatively fast and,
therefore, require faster than normal drying so that their moisture
content is low enough at the end of yellowing to prevent scalding in
the leaf drying stage. For conventional barns under these conditions,
he suggested that less tobacco be placed in the barn, and it may be
necessary to keep both the bottom and top vents open for all or most
of the yellowing period. For bulk barns, reducing the amount of
tobacco may cause uneven air flow through the tobacco, so starting
the ventilation process earlier than normal may be the preferred
option. Upper stalk tobacco harvested under dry conditions presents
the reverse situation. The problem is to keep enough moisture in the
leaves so that yellowing can be completed before the cells are killed
by excessive drying. Under these conditions, the drying rate should be
reduced by uniformly loading more tobacco in the barn and keeping
the vents closed during most or all of the yellowing period.
For the color-fixing and leaf drying stages, which last about 2 days for
normal ripe tobacco, drying rate is increased by increasing both
ventilation and temperature (Fig. 5.2). During this stage, enzymatic
activity and other cellular functions are inactivated by desiccation
and/or lethal temperatures, approximately 70% of the original green
leaf weight is lost, primarily as water, and much of the final chemistry
and color of the cured leaves are preserved (Peele, et al.,1995). The
temperature is increased about 1.1°C/hour from 35 to about 55°C
which is maintained until the lamina are almost dry. The 2-day period
is required for moisture removal to keep pace with the temperature
increases. Sufficient ventilation is provided to hold the wet bulb
temperature at 38°C for the first 24 hours and no higher than 41°C for
the remainder of leaf drying. Wet bulb temperature closely
approximates leaf temperature until the leaf is dry (Watkins, 1983).
Leaf cells break down and oxidative browning or scalding occurs at a
leaf temperature of 45°C, especially in the thinner leaves from the
lower stalk positions. Therefore, it is suggested that the wet bulb
should not exceed 41°C until the lamina are completely dry. Scalding
can occur when the dry bulb temperature exceeds 55°C if sufficient
moisture has not been removed from the leaves, thus another reason to
remove about 30% of the moisture during the yellowing stage
(Collins, 1983). This is why most successful curing schedules include
a period of constant dry bulb temperature at the end of leaf drying but
before advancing to the stem drying stage (Collins & Hawks, 1993;
Watkins, 1983; Fig. 5.2). In bulk barns, scalding is most likely to
occur in heavily loaded areas of racks or boxes which do not receive
adequate air flow.
In the stem drying stage, temperature is gradually increased from 55
to about 73°C, while ventilation is gradually decreased. Peele, et al.
(1995) stated that the midribs still contain large amounts of water after
leaf drying and high temperatures are necessary to overcome
evaporative cooling. The higher temperatures also reduce relative
humidity in the barn which increases the rate of stem drying.
However, Watkins (1983) suggested providing enough ventilation to
keep the wet bulb temperature at 43°C until the lamina are completely
dry to reduce the risk of scalding or sponging. Prolonged temperatures
above 75°C at the end of stem drying may increase the amount of
scorched, red tobacco (Collins, 1983; Watkins, 1983), especially for
some varieties (Hawks, 1972). In contrast to 'cherry red', which is
associated with high concentrations of nornicotine (Weeks, et al.,
1993) and disappears after the leaves are placed in the sun for about a
minute, scorched red tobacco often has a sweet off-type smell caused
by the caramelization of sugars at high temperatures (Flower, 1994).
In addition, Papenfus (1996) stated that stem drying temperatures high
enough to cause undesirable color changes do not result in faster stem
drying but instead remove valuable essential oils and associated
aromatic compounds from the cured leaf.
The causes and symptoms of major curing disorders, including those
discussed above, have been published by others (Walker & Stier,
1987; Collins & Hawks, 1993; Flower, 1994). The publication by
Flower (1994) contains photographs of nearly 30 curing disorders.

 Page 134
c
Current Curing Research in the USA
A major problem with the development and implementation of curing
in larger boxes is the difficulty in loading a consistent density of green
leaves into the box (W.H. Johnson & M.D. Boyette, personal
communication, NC State University). One possible solution would be
to slice or cut the green leaves into smaller pieces at harvest. Johnson
conducted research 25 years ago (unpublished) that demonstrated that
cut leaves packed into boxes more uniformly than whole leaves. Other
potential advantages of cut leaves are that they may be conveyed
pneumatically, leading to total harvesting and curing mechanization,
and possibly have a shorter curing time than whole leaves. Both labor
problems and the use of harvesting and curing mechanization continue
to increase in the USA, so Johnson has renewed his research efforts
with curing cut tobacco. In addition to the proposed advantages listed
above, preliminary results indicate that 40 to 50% more green weight
can be cured in boxes with cut than whole leaf, without affecting
visual quality and curing time. To date, however, cut cured leaves
have had slightly but consistently higher starch and lower reducing
sugar concentrations than whole leaves, probably the direct result of
bruising during cutting (Johnson, et al., 1957; Comber, 1977) and/or
relatively faster drying along the cut leaf edges during the yellowing
and early colorfixing stages. W.H. Johnson is currently investigating
cutting and curing techniques to correct this problem.
Based on the assumption that the major chemical changes take place
during yellowing, Johnson (1996) used infrared radiation to dry
yellowed tobacco lamina within 5 minutes compared to normally
cured leaf which required 24 hours or more to reach the same state of
dryness. The rapidly dried product was chemically and physically
similar to normally cured tobacco and also compared favorably in
smoke taste quality. However, under rapid drying conditions, reduced
percentages of polyphenols and nicotine were obtained, as well as
slightly higher levels of total reducing sugars, probably due to fewer
respiratory losses during rapid drying. Johnson (1996) suggested that
major considerations for future work are to rapidly dry midrib and
lateral vein materials, which have heat and moisture transfer
characteristics different from lamina, and, if successful, to explore
rapid drying of yellowed tobacco on a commercial basis.
d
Yellowing Chemicals
Tobacco growers in some major flue-cured countries, particularly the
USA and Canada, use ethephon (2-chloroethylphosphonic acid) sprays
1 to 2 days before harvesting or ethylene gas during the yellowing
stage of curing. The growers believe these chemicals allow greater
flexibility in harvesting immature tobacco, increase barn capacity by
reducing yellowing time and improve the color of cured leaves. The
chemicals are promoted as enhancers of senescence or yellowing of
tobacco leaves similar to that produced by the natural senescence
hormone, ethylene (Jones & Wilkinson, 1996). In some instances,
both chemicals are used on the same tobacco, particularly if it is
unripe (personal observation), although neither chemical is
recommended for use on immature tobacco. Tobacco purchasers are
concerned about the use of yellowing chemicals because ethephon use
has been reported to sometimes adversely affect cured leaf chemistry
(Dormir & Foy, 1976; Long, et al. 1974; Walker, 1977) and ethylene
gas reduced smoking quality of Canadian tobacco (Walker, et al.,
1985). It is possible that use of these products can mask the
immaturity of tobacco harvested under-ripe (Peele, 1994).
Results of several early field studies (Miles, et al., 1972; Dormir &
Foy, 1976; Walker, 1977) demonstrated that ethephon treatment on
mature tobacco increased yellowing rate in both the field and curing
barn. In most early research with ethephon, mature tobacco was
sprayed several days before harvest, allowing the leaves to yellow in
the field, and reductions in yield and visual quality sometimes
occurred (Miles, et al., 1972; Dormir & Foy, 1976). However,
improvements in visual quality have been reported for ethephon
treatment (Steffens & Alphin, 1970; Kittrell, 1983), with no effects on
yield (Kittrell, 1983).
The reported effects of ethephon on the direction and magnitude of
changes in the starch, sugar and nitrogeneous constituents of flue-
cured tobacco are highly variable, even in experiments where
adequate yellowing responses occurred in the field. Long and
Weybrew (1981) suggested that the variable results among
experiments were probably due to differences in climatic conditions
(especially temperature), cultural practices (especially N fertilization)
and rate of ethephon. In a 2-year study with upper stalk tobacco
harvested 24 hours after ethephon treatment (Gooden, et al., 1997),
results with ethephon appeared to be influenced by seasonal and/or
curing factors. Nontreated and treated tobaccos were yellowed for 72
and 48 hours, respectively. Grade and price indices were reduced by
ethephon treatment in the second year, but the physical parameters
measured were not consistently affected by ethephon in either year.
Reducing sugars were higher in ethephon-treated tobacco in the

 Page 135
second year and a greater proportion of the cured leaves were graded
as smoking leaf (H) in the first year. This study also included three
application and harvest dates for both treated and control tobaccos.
The first harvest was 24 hours after leaves of treated test plants
showed a yellowing response to ethephon; the second and third
harvests were made 10 and 20 days later, respectively. For the first
harvest, assumed to be physiologically mature leaves, smoke panels
preferred ethephon-treated tobacco, but the preference was for
nontreated tobacco as harvest was delayed. Peele (1994) reported that
smoke panel acceptability was less for overripe than ripe tobacco
treated with ethephon, while cigarettes made from overripe tobaccos
treated or not treated with ethylene gas were judged as acceptable.
However, panel acceptability of immature tobacco was not improved
by treatment with ethephon and/or ethylene gas.
Ethylene gas is injected into the curing barn during yellowing and its
proposed advantages, i.e. faster yellowing and better visual quality,
are promoted to be similar to those of ethephon. Rakitin, et al. (1976)
reported that an ethylene to air ratio of 1:1000 in small, wooden
chambers increased the yellowing rate of leaves of a Russian tobacco
variety, Ostrolist 1519. Green leaves exposed to ethylene had slightly
higher respiration rates, while cured leaves contained lower protein
and starch concentrations but higher concentrations of reducing
sugars. Walker, et al. (1985) found that ethylene gas generally
increased yellowing rate and buyer preference of Canadian flue-cured
tobacco, but darkened the base color of cured leaves and reduced
smoke quality. In a 3-year study using variety K-326, Goins, et al.
(1996a) evaluated the use of ethylene gas on tobacco harvested
immature or mature and found negligible differences in yellowing
time between ethylene-treated and nontreated leaves. Most differences
in leaf chemistry were associated with ripeness at harvest rather than
ethylene treatment. In addition, ethylene did not influence grade index
or smoking characteristics of cured leaves compared to the nontreated
or smoking characteristics of cured leaves compared to the nontreated
tobaccos, although an industry smoke panel did prefer ripe tobacco
over unripe tobacco (Goins, et al., 1996b). These workers concluded
that ethylene gas did not influence the chemical or subjective quality
of flue-cured tobacco in a consistent or meaningful manner.
Only two studies that included both ethephon and ethylene gas have
been encountered, and the results of both indicate that ethephon is
more effective than ethylene gas in reducing yellowing or total curing
time. In a 3-year unpublished study by M.J. Rogers (Virginia Polytech
Inst, Blackstone, 197779), use of the ethylene generator reduced total
curing time of variety Coker 319 an average of 11 hours compared to
the nontreated control. However, ethephon sprayed 24 or 7296 hours
before harvest reduced curing time by 21 or 35 hours, respectively.
Only when ethephon-treated tobacco was harvested 4 hours after
treatment (not labeled or recommended) did both products have
similar curing times (106 hours for ethophon and 105 hours for
ethylene). In three experiments with variety K-326, Peele (1994) used
ethephon alone, ethylene gas alone or both on the second large
priming (upper 14 to 16 leaves) of twice-primed tobacco harvested
underripe, ripe and overripe. Average yellowing times for the
nontreated tobaccos harvested at the three stages of ripeness were 74,
66 and 55 hours, respectively. Therefore, the first step in reducing
yellowing time is to allow the tobacco to ripen in the field before
harvesting. Ethephon used alone or in combination with ethylene gas
further reduced yellowing time by 7 to 24 hours, depending on
location and ripeness at harvest (Table 5.10). Use of ethylene gas
alone did not affect yellowing time in any experiment, and using both
products on the same tobacco did not reduce yellowing time more
than use of ethephon alone. However, in experiment 3, where the
underripe tobacco was very immature at harvest, none of the
treatments affected yellowing time. For the agronomic parameters
mea-
Table 5.10 Effects of ethephone and/or ethylene gas on yellowing time (in hours)
during curing (Peele, 1994).
Experiment 1 Experiment 2 Experiment 3
Yellowing treatment UR R OR UR R OR UR R OR
Control 84 71 65 68 61 49 70 66 51
Ethephon alone 60 47 44 52 49 42 70 54 37
Ethylene gas alone 84 71 65 68 61 49 70 66 51
Ethephon and ethylene 60 47 44 52 49 42 70 54 37
UR = underripe; R = ripe; OR = overripe.

 Page 136
sured under the conditions of this study, Peele concluded that the most
consistent positive effect of ethephon was a reduction in yellowing
time, while ethylene gas provided no measurable beneficial effects.
The reasons for the lack of yellowing response to ethylene gas in the
studies of Peele (1994) and Goins et al. (1996b) are not clear, but may
be related to use of higher N rates and more vigorous varieties than
those used in the studies conducted by Walker, et al. (1985) and
Rogers (1979, unpublished).
d
Marketing (See Also Chapter 10A of this Monograph)
The colors of cured leaves and their uniformity and intensity within a
lot of tobacco are important characteristics used by buyers to
determine its usefulness for specific blends (Campbell, 1982).
Therefore, leaves with undesirable colors (particularly green, black
and brown) are usually removed by farmers before offering the
tobacco for sale. In most countries except the USA, the cured leaves
of each priming are further separated into homogeneous lots based on
variations in leaf size, color, texture and degree of blemish, and then
bulked for several weeks before marketing. Minimum sorting is
practiced in the USA and the cured tobacco is usually marketed within
a week after being removed from bulk barns. In some southern
African countries, where labor is currently plentiful and inexpensive
and most of the crop is exported, a crop may be sorted into 60 or more
lots (i.e. grades) by the grower before being sold. In some countries
such as Mexico, green tobacco is purchased directly from the grower
and collectively cured by the buying company. However, most
fluecured tobacco is sold in the cured form in standardsized,
compressed bales weighing 40 to 100 kg, depending on the region or
country. In the USA, fluecured tobacco is currently marketed in burlap
sheets weighing about 100 kg each, but the use of bales weighing 275
to 450 kg is being investigated. If accepted as a practice, this will
change many aspects of preparing, marketing and processing
fluecured tobacco in the USA.
In most countries, tobacco is produced and sold under contract
between the grower and purchaser, with predetermined, negotiated
prices for the various grades. In a few large tobacco-producing
countries, annual production is controlled by government regulation
or farmer-buyer negotiation, and the tobacco is sold in an auction
system with (USA and Canada) or without (Zimbabwe) minimum
support prices for most usable grades. In the USA, tobacco not sold to
commercial buyers is purchased for price support by a grower-owned
cooperative and sold later to domestic and foreign buyers, usually
when supply is limited. Some marketing systems, such as that in
Zimbabwe, have been substantially mechanized. However, a great
deal of manual labor is still required to unload and present the tobacco
for sale, remove it from the sale area and to load and transport it to the
buyer's processing facility.
The quota system used in the USA limits the amount of excess
production that can be sold in the marketing season. Therefore, excess
tobacco is often stored on the farm until the next season. To reduce
spoilage, tobacco to be stored should not contain more than the
minimum moisture content that will allow it to be handled without
breakage and shattering. Stored tobacco is also subject to damage by
the tobacco moth and the cigarette beetle, particularly in warm
climates, and both pests are difficult to control after the tobacco has
been packaged or bulked. Southern (1996) listed several suggestions
to minimize insect infestations of stored tobacco and appropriate
treatments should infestations occur.
Labor Requirements
The manual labor required to produce and prepare fluecured tobacco
for sale varies greatly around the world, depending primarily upon the
level of mechanization used for transplanting, sucker control,
harvesting and preparation of the leaves for curing and marketing.
However, harvesting and the preparation of leaves for curing and
marketing are usually the most labor intensive tasks in tobacco
production in most countries. In the USA, the total manual labor
required ranges from 150 to 300 man-hours per hectare, depending
primarily on the level of mechanization used for harvesting and
loading/unloading of bulk barns. These estimates would be expected
to be considerably higher in countries using less mechanization.
The Future
Major changes in tobacco production over the last 50 years have
occurred in response to economic factors affecting production
efficiency. Additional changes, including some for the tobacco
program in the USA, will have global implications. Imaginative
research in management, harvesting and curing technology will
continue to improve production efficiency of quality

 Page 137
fluecured tobacco in the future. Advances in breeding and
biotechnology will undoubtedly provide greater tolerances to the
major diseases and insects, perhaps reduce the adverse effects of
weather extremes on yield and quality, and eliminate and/or reduce
undesirable chemical constituents in cured leaves. The requirement
for quality tobacco for quality products will not change, although
buyer preferences for some styles of tobacco may change as cigarette
manufacturers' technology improves and consumer demand changes.
Thus, tobacco agronomists will continue to be challenged to
incorporate and maximize the effects of positive changes into our
industry. In closing this chapter, however, it is appropriate to expand a
statement made by Chaplin and Miner in 1980,
'We can afford to adopt only those new cultural practices that improve the
grower's production efficiency while preserving the flavors, aromas,
chemistries, and physical properties of cured leaves desired by the
manufacturer and consumer'.
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5B
Light Air-Cured Tobacco
G.K. Palmer and R.C. Pearce
University of Kentucky
Lexington, Kentucky, USA
Introduction
Light air-cured tobacco is common in many locations all over the
world. Production practices vary depending on local weather patterns,
socio-economic status of the growers and the degree of
industrialization of the region. Even in developed countries, light air-
cured tobacco production is a labor-intensive enterprise when
compared with other crops. The common factor is that curing is
primarily without artificial sources of heat and humidity. Because air-
cured tobacco is not cured under a prescribed set of temperatures and
humidities, the end product may differ considerably from one location
to another and from year to year. The two most common types of light
air-cured tobacco are burley and Maryland.
Maryland tobacco refers to the relatively light bodied, mild, air-cured
tobacco grown on the sandy coastal plains of southern Maryland,
USA. Maryland tobacco has a relatively small, but loyal niche market
in European cigarette manufacturing. Some blended cigarettes contain
small amounts of Maryland tobacco.
Burley tobacco comprises most of the light air-cured tobacco. Burley
tobacco, as we know it today, originated in Southern Ohio and
Northern Kentucky, USA in 1864. Because of this, the light air-cured
tobacco produced in Kentucky and the surrounding states set the
standard of quality for burley tobacco. Burley tobacco growers in
other parts of the world strive to produce a similar style of leaf, but
often find it difficult to duplicate the growing and curing conditions
necessary to produce a full flavor burley. This milder burley type leaf,
often called filler tobacco and valued for its filling power and open
structure, accepts flavoring compounds well. Most burley type
tobacco becomes a component in the manufacturing of blended
cigarettes.
Soil Factors
Light air-cured tobacco production utilizes a wide range of soil types
around the world that influence the character of the cured leaf. Soil
factors with an impact on tobacco production include soil texture,
structure, fertility and landscape position. Management practices such
as tillage and fertilization modify the crop response to these factors
and influence the style of tobacco produced.
Soil texture is an inherent soil property that is not subject to easy
manipulation. The conventional wisdom is that sandy soils produce
very light-bodied open tobacco, and clay soils are more likely to result
in heavy-bodied tight leaf. Most air-cured tobacco grows on
intermediate loamy textured soils.
The relatively coarse textured soils of Southern Maryland tend to
produce light-bodied tobacco with an open leaf structure and the mild
flavor typical of Maryland tobacco. In one study Maryland tobacco,
grown on soils ranging from fine sand to loam textures, produced
higher yields, total alkaloids and total N with lower filling capacity on
loam soils when compared to that grown on coarser textured soils
(Mulchi, 1985). Visual quality was lowest for tobacco grown on fine
sand. In general, loams and sandy loams are the best soils for the
production of Maryland tobacco (McKee & Conrad, 1994).
Burley tobacco production is common on both coarse and fine
textured soils in various parts of the world. Much of the burley
tobacco produced in Latin America and Africa grows on sandy loams
and loams and tends to be 'filler' type burley. Cured tobacco in these
areas has a light-body, open leaf structure and a neutral character with
little impact on the smoke (Glass, 1983). Conversely, in areas of
heavier silt loams and silty clay loams the tendency is to produce a
medium-bodied tobacco that imparts a strong burley character to the
smoke. The tobacco grown in the US burley belt is an exception to the
rule that heavy soils produce tight, lifeless leaf. Although most burley
grows on silt loam soils with clay subsoils, the cured leaf is relatively
open in structure and accepts flavorings well.
A soil with good internal drainage produces the best burley (Sims,
1993) and Maryland tobacco (McKee & Conrad, 1994). Sandy soils
generally have good

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drainage because of their coarse texture; however, heavier soils must
have a strong stable soil structure, particularly in the subsoil, to
maintain good drainage. A fine textured soil with poor structure and
drainage will percolate water very slowly and have a greater tendency
to become saturated in the root zone. This will result in reduced root
growth and increase the potential to produce thin-bodied leaf.
Maintenance of soil structure is essential to continued production of a
high yield of good quality light air-cured tobacco.
With its high biomass production, tobacco removes relatively large
quantities of nutrients from the soil. Soil fertility refers to the
capability of the soil to supply these nutrient elements to crops. Very
few soils are fertile enough to supply the needs of a high yielding
tobacco crop without supplemental fertilization. Typically, sandy soils
are more difficult to manage due to the relatively low capacity to store
nutrients in the root zone. Finer textured, deep soils are generally
more fertile because of their capacity to retain nutrients in forms that
are not subject to leaching loss, yet are readily available to plants.
Selection of flat to gently rolling landscapes for air-cured tobacco
production prevents excessive loss of soil due to erosion. Most burley
tobacco production in the USA is on flat land without ridges or beds.
This, coupled with intensive tillage practices, makes tobacco ground
very susceptible to soil erosion. Soil erosion can, in just a few years,
destroy the productivity of the soil. Steep-sided slopes hasten the loss
of productivity. Other landscape positions can cause production
problems. Depressional areas often produce poor tobacco due to water
logging. Bottom lands near streams and rivers can have drainage
problems but, where well drained, bottom lands produce excellent
tobacco crops. Well drained upland sites, ridge tops or gently rolling
side slopes, with deep soil that has sufficient water capacity
consistently produce the best tobacco crops. The coastal plain region
is flat to gently rolling and well suited for the culture of Maryland
tobacco.
Site Selection and Tillage Practices
Site selection for tobacco production 1 to 2 years prior to growing the
crop provides ample time to test for chemical properties and make
amendments. Besides soil drainage, good air drainage is also
advantageous. Sites with limited air drainage may be more prone to
certain diseases like blue mold or target spot. However, factors other
than soil conditions, such as proximity to irrigation or curing facilities,
often dictate site selection.
Crop rotation is necessary to maintain soil tilth and reduce disease
incidence. An ideal rotation for burley tobacco is tobacco at the same
site for 2 years and then grass sod for 4 years. However, due to land
limitations, the same site may have continual tobacco production or
limited rotation. Continuous tobacco production can result in severe
soil degradation due to intensive tillage and the high nutrient demand
of tobacco. Proper fertilization replaces depleted nutrients, but
structural degradation continues. Structurally degraded soils tend to be
cloddy and puddle easily. Such soils may require more intensive
tillage to produce a 'suitable' seed bed, thus perpetuating a destructive
cycle. In many parts of the world the lack of sufficient land area for
adequate rotation is a serious impediment to air-cured tobacco
production. Proper rotation intervals reduce the severity of some soil-
borne diseases like black shank and black root rot, although some
rotational crops, like legumes, can maintain damaging levels of soil
pathogens.
Site preparation generally begins with a primary tillage operation. In
developed countries, cultivation of the site usually involves turning of
the soil with a moldboard plow in late winter or early spring. This
operation kills any winter vegetation and buries surface residues so
they will decompose. Sealing plowed soil with a heavy drag levels the
soil and aids in organic matter decomposition. Plowing a large
quantity of residue under near setting time contributes to a disorder
commonly known as organic matter toxicity. Organic matter toxicity
causes yellowing and stunted growth and is the result of a toxic
accumulation of nitrite from the rapidly decomposing organic matter
(Hamilton & Lowe, 1981). Plowing sods and heavy cover crops at
least 6 weeks before setting reduces the chances of organic matter
toxicity.
Working the field site several times with a disk harrow or similar
implement before setting produces a good bed for transplanting. Field
preparation provides a means of incorporating any fertilizer,
herbicides, fungicides and insecticides needed. The use of power
driven tillers leads to more rapid destruction of soil structure.
Intensive cultivation after transplanting contributes to structural
degradation and soil erosion. Cultivating for weed control may be
essential. However, deep cultivation damages roots and brings viable
weed seeds to the surface.
Minimal or no-tillage methods of production reduce soil loss but
comprise a small amount of the total

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production. No-till production methods generally result in greater
reliance on chemical methods of weed control. Modified transplanters
can successfully establish seedlings in chemically killed sods and
cover crops (Phillips & Zeleznik, 1989). Adequate control of weeds in
no-till tobacco production allows the tobacco the chance to yield
comparably to that of conventionally grown tobacco (Pearce &
Zeleznik, 1996). Inconsistent weed control limited adaptation of no-
till in the past. New herbicide chemistry recently approved for use on
tobacco makes no-till tobacco more feasible.
Developing nations utilize hand or animal power for tillage.
Nevertheless, tillage of tobacco ground is intensive in these countries
compared to other crops. Clearing ground for tobacco production is
common in some regions. After several years of tobacco production, it
is transferred to newly cleared land and the abandoned area
regenerates. Land for this type of slash and burn culture is becoming
scarce. In some places it is prohibited by regulations. Production of
light air-cured tobacco on top of low ridges or beds, formed following
any secondary cultivation and just before transplanting, is common in
some countries.
Fertilization Practices
Fertilization practices for light air-cured tobacco vary depending on
the type of product desired. Producers of Maryland and filler burley
tobacco use low rates of nitrogen in the 70 to 90 kg/ha range (McKee
& Conrad, 1994). Burley tobacco growers in the USA typically apply
200 to 300 kg N/ha, while in Kentucky, N rates approaching 300 to
400 kg N/ha are common. Ammonium nitrate and urea are the
primary sources of N with sodium nitrate more commonly reserved
for side dressing. As a result of heavy nitrogen applications, burley
growers must apply large quantities of lime to offset the acidity
produced by nitrogen fertilizers. Maryland lime recommendations call
for liming to a pH of 5.5 to 5.6 (McKee & Conrad, 1994). The
recommendation for burley tobacco in Kentucky is to lime to pH 6.4
to 6.6 (Anon, 1996). The reason for liming is to insure that after
fertilization the pH does not drop below 5.4 where manganese (Mn)
toxicity can become a problem.
Manganese toxicity, considered to be the most significant nutritional
disorder of burley tobacco in Kentucky, generally appears 2 to 4
weeks after setting. Tobacco develops poorly with interveinal
chlorosis between veins that remain dark green. A rescue treatment of
1100 kg/ha fine lime improves soil pH and helps the tobacco recover.
However, prevention is the key to reducing losses from Mn toxicity.
Agricultural grade limestone applied at least 6 to 12 months before
planting tobacco produces the best results.
Recommendations for phosphorus range from 0 to 200 kg P/ha based
on soil test levels. Limited root growth in the spring under cool, wet
conditions compounds phosphorus deficiencies. The symptoms of
deficiency include stunted growth and occasionally the appearance of
small, shiny, metallic looking spots on the lower leaves of young
plants. This condition is usually temporary and normal growth
resumes eventually with little or no impact on cured leaf yields. The
most commonly used source of phosphorous is triple super phosphate
with ordinary super phosphate also used in some regions.
Potassium is well known for its effects on improving the burning
qualities of cured leaf. Light air-cured tobacco, because of its large
biomass production and high potassium content, has a high
requirement for potassium. The symptoms of deficiency are first
chlorosis and eventually necrosis of the leaf margins of older leaves.
Affected leaves will be thin and trashy at harvest. Potassium
deficiency can reduce cured leaf yields by as much as 800 to 1000
kg/ha. Soil tests for available potassium provide assessment of needs
that range from 0 to 400 kg K/ha. The preferred source is potassium
sulfate. The cost of fertilization tempts tobacco growers to use the
cheaper muriate of potash source. Muriate of potash costs half as
much per unit of K as potassium sulfate, however the use of muriate
can lead to the accumulation of unacceptable levels of chloride and
impair the quality of the cured leaf.
Trends in recent years in the USA are toward the use of customized
blends, based on soil tests, that provide only the nutrients needed for
the current crop year. A practice that was common in the past, and still
in use today, is to apply a prescribed quantity of blended fertilizer,
regardless of the crop need. The most common 'recipe' in recent years
called for 2240 kg/ha of 5-10-15 fertilizer plus a supplemental
application of nitrogen. Such practices are inefficient and may result
in the over-or under-application of some nutrients.
In the USA, the majority of the fertilizer applications are made prior
to transplanting with incorporation into the top 7.6 to 10.2 cm of soil.
Some growers sidedress nitrogen at the first or second cultivation to
improve fertilizer efficiency and to replace nitrogen lost by leaching.
In developing regions where fertilizer is scarce and expensive,
growers are more likely to apply lower rates of fertilizer in bands or to
individual plants to improve efficiency. Banding or other forms of fer-

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tilizer placement can improve fertilizer efficiency by as much as 50%
compared to broadcast applications (Sims, et al., 1994).
Micronutrient deficiencies in tobacco are relatively rare and usually
occur in isolated pockets. Most micronutrient deficiencies occur as a
result of pH imbalances. The only micronutrient recommended for use
on burley tobacco in Kentucky is molybdenum (Mo) when the pH is
less than 6.4. Transplant water solutions of Mo are easy and simple to
apply. Sims (1980) reported that Mo fertilization in Kentucky resulted
in a significant yield increase on about 50% of the sites tested. Yield
increases ranged from 225 to 900 kg cured leaf/ha. Precise
micronutrient fertilization is necessary due to a narrow window
between sufficiency and toxicity.
Transplant Production
The majority of burley tobacco transplants production in the USA is in
containerized plant production systems. The most commonly
employed system is the float system, in which transplants grow in
polystyrene flats (trays) floating on beds of nutrient solutions. The
float system has been in use since the early 1990s yet comprises over
70% of all burley transplants production. The cost is somewhat higher
than for conventional transplant beds, but for many growers the
advantages of the system outweigh the costs. The main advantage
cited by most growers is the ability to easily handle and transport
groups of plants instead of having to pull individual plants from a
conventional bed.
Excessive fertilization can lead to salt damage in young seedlings and
to increased disease susceptibility in older plants. The nitrogen
concentration in the float water should not exceed 125 ppm and the
preferred range for burley is 75 to 100 ppm N. Commonly used
fertilizers are 20-10-20, 20-9-20, and 15-4-15. These ratios provide
the proper balance of nutrients for good transplant growth. The use of
urea-based fertilizers in the float system produces nitrites that are
toxic to plants.
Seeds, pelletized with diatomaceous earth to increase size and
uniformity, allow for singulation and automated handling of seeds.
Seeding begins 6 to 8 weeks prior to transplanting, depending on light
and temperature. A peat-based soil-less mix placed in the trays wicks
moisture and nutrients from the float bed. Producers utilize
greenhouses covered with polyethylene film or unheated outside float
beds. Outside beds are subject to damage from rains and cold
temperatures.
The float system dramatically increases the long distance transport of
tobacco transplants in the USA. Suitable precautions are necessary to
prevent the spread of diseases by infected transplants.
A condition known as spiral root is common for seeds germinated in
the float system. Spiral root is a result of media over-saturation. The
extreme moisture in the float system makes for an environment that is
more conducive to disease development. Plants produced in float
systems appear to be more susceptible to chill injury than
conventionally produced plants. Though the transplants usually
recover from this chill injury, problems with lateral suckers from the
base of the stem increase as a result of bud damage.
Conventional tobacco beds are still common in many areas of the
world. Fertile, well drained soils are preferable. The site should be
free from shade and located near a source of clean water. A fine seed
bed is important to insure the best start for seedlings. A broad
spectrum biocide used to fumigate the site kills weed seed and
pathogenic organisms. A gas-impermeable covering prevents rapid
dissipation of the fumigant and protects from recontamination. A mix
of raw tobacco seed and sand or fertilizer allows a uniform
distribution of seeds. Straw or other mulch helps hold moisture and
provide insulation where needed. A cotton, polypropylene or
polyethylene cover permits light to pass but retains heat. The
provision of adequate moisture promotes good plant growth.
One of the issues facing growers of conventional plant beds is the
potential loss of the fumigant, methyl bromide, due to environmental
concerns. Few alternatives exist for use on tobacco plant beds.
The ideal burley tobacco transplant is 10 to 15 cm in height with a
stem diameter of 8 to 10 mm. A healthy burley transplant has a pale
green color. Dark green colors indicate a plant that is tender and may
have more difficulty surviving in the field.
Plant Population
Plant population can have a significant impact on economic return per
unit area, plant size parameters, yield and quality. Economic and
quality considerations should determine the plant population.
However, producers often base plant population decisions on
tradition. Manual transplanting often has an inadvertent influence on
plant population. Limiting factors, such as land availability and labor,
are the best indicators for determining plant population. If labor is
limiting, a decrease in plant population produces more

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cured leaf weight per unit handled. If land is the limiting factor, an
increase in plant population produces more cured leaf weight per unit
area. If quality is significantly affected by a high or low population,
plant population should be shifted to improve quality.
In the USA before 1971 a government control program limited burley
production based on land area. Limits (quotas) are now based on
cured leaf weight. Production gradually shifted to wider plant spacing
as labor availability decreased. Current recommendations call for
plant populations to range from 15 000 to 18 000 plants per hectare.
However, spacing ranges from 40 to 60 cm within a row and 90 to 120
cm between rows. Maryland tobacco benefits from slightly wider
spacing with recommended populations ranging from 12 000 to 15
000 plants per hectare. Row spacing ranges from 90 to 107 cm with
spacings of 45 to 90 cm within a row. Producers in many areas of light
air-cured tobacco production would benefit from careful consideration
of optimum plant population (Isaacs, 1993; McKee & Conrad, 1994).
Weed Control
Weeds compete with light air-cured tobacco for water, nutrients and
space just as they do with all types of tobacco. Climbing weeds may
physically damage the tobacco before or during harvest. Mechanical
and chemical controls significantly reduce the loss of yield and quality
associated with weed growth. Whether by hand cultivation or motor
driven cultivators, a shallow cultivation is sufficient to disturb weed
root systems or shear the weed stem just below the soil level. A deep
cultivation is not necessary and may damage tobacco root systems or
pull untreated soil to the surface, allowing regrowth of weeds.
Cultivation during dry conditions exposes soil moisture and promotes
evaporation. Cultivation with machinery when soils have high soil
moisture contributes to soil compaction.
The number of cultivations required depends on weed species and
population, weather conditions, efficiency of cultivators, herbicide use
and how well the tobacco competes with the weeds. The light air-
cured tobacco produces a moderate amount of shading quickly. By 6
weeks tobacco is capable of shading most weeds that regrow later in
the season. Exceptions include perennial weeds that may have
established root systems or other structures that have the potential to
grow rapidly. Yellow nutsedge (Cyperus esculentus L.), which
reproduces primarily from underground tubers, can re-establish
quickly and crowd out newly transplanted tobacco plants. Although
cultivation may disturb or kill a nutsedge plant, the death or severance
of the mother plant from the tubers stimulates sprouting and increases
the weed problem. Annual climbing weeds such as morning glory
(Ipomea sp.) and honeyvine milkweed (Ampelamus albidus (Nutt.
Britt.) may grow sufficiently to intertwine tobacco leaves, causing leaf
breakage and loss.
Chemical weed control is common in light air-cured tobacco
production. An ideal herbicide selectively controls weeds without
damaging the tobacco. It persists long enough to prevent loss of yield
or quality from weed pressure without causing damage to any
subsequent crop. Although ideal herbicides don't exist, several
chemicals successfully control weeds in light air-cured tobacco with
tolerable effects on the crop.
Pendimethalin (Prowl®) is a dinitroanaline herbicide applied
preemergent or preplant incorporated for control of most annual and
certain broadleaf weeds (Thomson, 1993). Pendimethalin controls
weeds by inhibiting cell division of roots or shoots emerging from a
germinating seed. Incorporation to a depth of no more than 5 cm
prevents loss from photodecomposition and volatility, places the
chemical in the weed seed zone and allows room for adequate root
development below the treated zone. Photodecomposition and
volatility account for minor losses in activity compared to some
herbicides. Rapid incorporation is, therefore, not necessary. A high
application rate, deep incorporation or soils with a high clay content
increase the potential for root damage. Since little or no uptake of the
chemical occurs, damage is primarily in the treated root zone.
Pendimethalin injury induces root swelling and prevents secondary
and tertiary root development. Low rainfall during the growing season
reduces the breakdown of pendimethalin, increasing the persistence
and the potential damage to cover crops or other rotational crops.
Benefin (Balan®) and isopropalin (Paarlan®) are similar herbicides
used to a lesser extent in tobacco production. They vary in their
persistence, photodecomposition rate and crop safety (Thomson,
1993).
Clomazone (Command®) is an isoxazolidinone herbicide applied
preemergent or preplant incorporated for control of most annual
grasses and certain broadleaf weeds (Thomson, 1993). Clomazone is a
chlorophyllinhibiting compound. Weeds germinate, take up lethal
doses of the chemical and turn white before dying. Some Amaranthus
species tolerate clomazone, therefore other means of control are
necessary where infestations of amaranths occur. Although new
formulations of

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clomazone volatilize less readily, damage to susceptible plants near
areas of application is possible. Incorporation reduces drift. Light air-
cured tobacco tolerates clomazone, but a few white leaves are
common. They recover quickly with no lasting damage to yield or
quality. Clomazone persists for a long period of time in soils,
presenting a threat to susceptible rotational or cover crops.
Pebulate (Tillam®) is a thiocarbamate herbicide applied preplant
incorporated for control of nutsedge, annual grasses and certain
broadleaf weeds (Thomson, 1993). Volatilization occurs quickly,
requiring immediate incorporation to reduce loss of activity. Pebulate
leaches easily in sandy soils and with heavy rainfall. Microbial
breakdown reduces persistence, potentially allowing regrowth of
weeds before harvest. Good activity on nutsedge makes it an ideal
chemical in areas with high nutsedge infestation. A tank mix with
other herbicides provides a broader spectrum of weed control.
Pebulate has excellent crop safety. Tobacco tolerates pebulate by rapid
breakdown of the chemical. However, under conditions of rapid
transpiration newly transplanted tobacco plants with extensive root
systems absorb excessive pebulate. Plants develop strap-like leaves
and/or bud damage. Pebulate damage is rare.
Napropamide (Devrinol®) is a propionamide herbicide applied
preplant incorporated or preemergent for annual grasses and certain
broadleaf weeds (Thomson, 1993). Napropamide is highly persistent,
lasting up to 12 months in most soils, and can damage rotational or
cover crops. Banding of napropamide over a tobacco row before weed
emergence can aid weed control. However, delaying application
increases the length of persistence. A tank mix with other herbicides
provides a broader spectrum of weed control.
Sulfentrazone (Spartan®) is a new soil-applied herbicide for selective
control of broadleaf and certain grass weeds (Anon, 1993). It is
especially effective on morning glories and nutsedge. Sulfentrazone
controls weeds through root and shoot uptake and subsequent
membrane disruption. Weeds turn necrotic and die after emerging
from the soil. A tank mix with other herbicides improves grass
control. Excessive application rates or incorporation of sulfentrazone
may cause leaf stunting and tissue breakdown on lower leaves of
tobacco.
Annual and perennial grasses in tropical and subtropical regions
present an extra challenge for tobacco producers. In countries where
selective grass herbicides are registered, they could be used to
improve weed control.
Topping
Before the development of effective sucker control chemicals, burley
and Maryland tobacco were topped late or just prior to harvest
primarily to prevent sucker growth (Anon, 1954). Other methods
included topping when two-thirds to three-fourths of the plants were
in bloom and suckering once or twice before cutting. Leaving the top
two suckers until just before harvest prevented other sucker growth.
Early topping in general produces a darker, heavier-bodied tobacco,
while late topping produces the opposite. Leaves of any plant serve as
a store for seed production. As blooms develop, energy stored in the
leaves moves to the area of highest demand, the seed head. Removing
the seed head in a timely manner prevents excessive loss from the
marketable leaves.
Since the topping stage affects yield, quality and nicotine content, the
method should fit the style of tobacco desired. In burley regions where
stalk-cut tobacco cures under good to ideal conditions, topping at 10
to 25% bloom produces the best quality and yield. Both bud topping
and late topping reduce yield but affect the quality differently. Ideal
topping for stalk cut Maryland is at 60% bloom. Stalk-cut burley
curing rapidly under less than ideal conditions may produce
undesirable quality if topped too early. The same applies to primed
burley and Maryland tobacco. Topping height depends not only on the
desired result but also on the sucker control method. Low topping
reduces labor but removes true tip leaves. Since a tobacco plant
supports a specific number of leaves, topping high does not insure
increased yield. Ideal leaf number varies but should range between 18
and 26 leaves. Length of the top leaf after topping varies but ranges
from 20 to 35 cm (Palmer & Calvert, 1993; McKee & Conrad, 1994).
Sucker Control
With the loss of apical dominance (normally by topping), laterals
shoots, or suckers, begin to grow. Suckers, like inflorescences, rob
leaves of valuable nutrients. Sucker control is essential for high yield
and quality. Hand suckering is one option, but chemical growth
regulators are more commonly used. However, in situations where
topping occurs after significant blooming begins, manual sucker
removal is essential before application of sucker control chemicals.
Sucker control chemicals fall into three categories:

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(1) systemics cnemicals absorbed by the plant and translocated inside
the plant,
(2) contacts chemicals that must come in contact with the sucker to
achieve control, and
(3) local or contact systemics chemicals that need to contact the
sucker bud where absorption occurs.
These chemicals differ in application methods, activity and residue
(Bruns & McKee, 1985; Palmer, et al., 1986).
Maleic hydrazide (MH) is a systemic chemical used for sucker control
in burley and Maryland tobacco in certain regions of the world, most
notably in the USA (Thomson, 1995). The most common type is the
potassium salt formulation used since the late 1970s. Removing
existing suckers reduces sucker control problems. This is especially a
concern in varieties of burley that initiate suckers quickly and in
Maryland tobacco where topping time may occur after sucker
initiation. Failure to remove existing suckers prior to application
encourages repeat applications that increase residue in cured leaf.
Application of MH within 24 hours after topping produces the best
results. Avoiding the hottest part of the day improves uptake and
control. Small leaves left after topping may develop slowly. MH
retards the spread of leaves smaller than 20 cm. A sufficient volume
of water (375 to 475 L/ha) should provide adequate coverage of the
top one-third to one-half of each plant. A reduced volume will
compensate for small tobacco and low plant populations. Current
research suggests that a coarse spray applied with low pressure (1.4 to
1.7 × 105 Pa) performs better than a fine spray mist, especially under
hot, dry conditions. MH-treated tobacco often yellows after treatment,
making assessment of maturity difficult. Other indicators must be
used to judge maturity when MH is used. Control lasts approximately
3 to 4 weeks depending on weather conditions and nutrient levels in
the plant. Rainfall within 6 hours after spraying requires a full
reapplication. However, after 6 hours and before 12 hours elapse, a
half rate is sufficient (Palmer & Calvert, 1993; McKee & Conrad,
1994).
The contacts contain fatty alcohols as their active ingredient. Fatty
alcohols must contact the sucker bud to achieve control (Thomson,
1995). Contacts kill small suckers rapidly, therefore fatty alcohols are
not susceptible to rainfall after 1 hour passes. Removal of suckers
greater than 2 cm in length improves control. Fatty alcohols work best
before sucker initiation. Applications at bud stage and before topping
or within 2 days after topping are effective. Arrangements of three
nozzles directed over each row are common on power sprayers. A
direct spray from a manual sprayer also provides good coverage and
control. Low pressure sprayers are sufficient for application of fatty
alcohols. A coarse spray or stream applied to the top of the plants
provides good coverage of leaf axillary buds. Leaves of tobacco plants
form a natural funnel that directs the spray down the stalk. Control
diminishes where plants are not upright. Straightening plants prior to
application could improve control. High rates of fatty alcohol and hot,
humid conditions promote leaf drop. Fatty alcohol efficacy lasts for 7
to 10 days and requires reapplication for season-long control. The
desired time between topping and harvest will determine the number
of reapplications required.
Fatty alcohols alone or in combination with other sucker control
chemicals are beneficial for improving uniformity in tobacco with an
irregular bloom stage. By topping and treating only those plants that
are in the desirable stage, uniformity improves. Repeating the process
in 7 days continues to improve uniformity if needed. However,
retreatment of initially treated plants is necessary when fatty alcohols
are used. MH as a final treatment or fatty alcohols applied as
sequential treatments controls suckers for the rest of the season.
Applications of MH too close to harvest increases undesirable residue.
Local or contact systemics are dinitroanaline chemicals that fall in the
same class as some herbicides (Thomson, 1995). Two examples are
flumetralin (Prime+®) and butralin (Butralin®, Tamex®). These
formulations control suckers in the same manner as they control
weeds, by interrupting cell division at the growing point. Since uptake
is minimal the chemical must come in contact with all axillary buds. A
spray mix may miss the growing point of suckers longer than 3 cm,
reducing control. The most common application method consists of
pouring a 2% solution to the top of the stalk. Volume will depend on
the size of the tobacco plants, but will be approximately 20 ml per
plant. Precise application is necessary to insure complete axil contact
and to prevent excess soil exposure. Residual chemicals on the soil
can damage cover crops and rotational crops. Poor sucker control is
common on plants that are not completely vertical. Misses often occur
in the top of the plant due to imprecise application. Any suckers
missed by the local systemic chemicals grow unimpeded. This could
include ground suckers stimulated by topping but that are large
enough for the sucker control mix to miss as it travels down the stalk.
Good coverage insures good control for a 6 to 8 week

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period of time. The length of control is more than sufficient to provide
the length (time) of control needed in either burley or Maryland
tobacco.
Combinations of reduced rates of MH and either of the dinitroanaline
type chemicals may offer advantages over MH alone, while reducing
the negative impact of each chemical. By reducing MH rates by a half
or three-fourths of the normal rate and by limiting local systemics to
1% of the final solution, a combination results with the best activity of
both chemicals.
Mechanical applications with high clearance sprayers equipped with
solid cone, coarse nozzles provide sufficient coverage and economical
application for large tobacco fields. Backpack sprayers similarly
equipped are equally acceptable. Besides excellent sucker control and
yield, a combination offers the true systemic activity of the MH and
the rain safety (wash off) and extended control of the dinitroanalines.
Good sucker control should reduce the tendency to over-apply or
reapply, which increases MH residue. Reducing the rate lowers MH
residue and decreases the cover crop damage associated with the local
systemics. Variation of the rates may be necessary depending on
production practices that may influence sucker pressure.
Harvesting
Maturity and ripeness are terms used frequently to describe the
appropriate time to cut tobacco. However, both terms are somewhat
ambiguous and often mean different things to different people. Since
light aircured tobacco must depend on natural curing conditions,
selection of the right time to harvest tobacco can influence the cure.
Although time after topping is the most common criterion used to
judge harvest, many factors influence true maturity or ripeness of a
crop. Time after transplanting is not a good criterion due to seasonal
variation. Weather has a primary influence affecting nutrient uptake
and active growth of tobacco. Other factors such as soil type, pH and
fertility influence maturity.
Plant characteristics such as color, stalk hardness and leaf firmness do
signal maturity but are not clearly defined (Massie & Smiley, 1974;
McKee & Conrad, 1994). A plant's normal green color degrades to a
lighter color as physiological maturity occurs. This may be subtle or
different depending on varietal influence. For example, varieties like
the burley variety, TN 86, or MD 609 grow with a light green color
initially. Stems and stalks of burley tobacco do turn a noticeable light
cream to white color at maturity, making assessment of maturity
easier in burley than Maryland tobacco. Stalk hardening begins as
floral initiation occurs and may not be a good indicator of the best
time to harvest. Delaying topping till full bloom allows the stalk to
harden before topping. This would eliminate stalk hardness as a useful
tool for determining maturity under these circumstances. A tobacco
leaf that breaks easily when bent signifies maturity and is a relatively
easy and nondestructive test. Leaf firmness is easy to evaluate but
may also vary due to moisture stress or time of day.
Harvesting light aircured tobacco is labor intensive regardless of the
method used. Depending on the location, light aircured tobacco
harvesting is by whole stalk cutting, leaf removal or priming and a
combination of the first two methods. The method used may depend
on several factors including labor availability, production practices,
curing facilities, curing conditions and the style of tobacco desired.
In the USA, whole stalk harvesting is almost the exclusive method
and it is becoming increasingly popular in countries where other
methods predominate. Cut tobacco left standing in the field to wilt is
susceptible to sunburn, which reduces quality. Significant water loss
during the field wilting period helps reduce labor demands and leaf
breakage. Maryland tobacco is more susceptible to sun damage than
burley, and field wilt should be minimal to reduce damage. However,
Maryland tobacco is more brittle at harvest and may suffer less
damage if allowed to field wilt for a short time. Sun intensity and
temperature in some locations prevent the use of field wilting as an
option. If tobacco sunburns, an extra 2 to 3 days in the field will
bleach the damaged areas, improving quality. However, quality still
suffers. Extending the time in the field exposes the tobacco to
potential rainfall in some locations. Rainfall can splash soil onto
leaves, further reducing quality or physically damaging more mature
leaves.
Priming of bottom leaves before stalk cutting to prevent deterioration
is common in some areas. Priming of leaves as they ripen or just the
bottom leaves of the plant before stalk cutting is also common in
many areas. Removal of leaves as they reach maturity is critical for
producing high quality tobacco. The leaf maturity characteristics
described above are useful in assessing maturity. Style desired may
differ, but in general leaves that are over-mature will produce a thin-
bodied leaf, and immature leaves may produce green tobacco when
cured.

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Curing
Moisture loss and curing begin at harvest. Primarily harvesting is by
hand with few exceptions, including mechanical leaf and stalk
harvesters in Japan and other locations (Ohori, 1996). Curing,
however, is more than drying of the leaves. It involves complex
physical and chemical changes that occur as tobacco leaves slowly
lose moisture.
There are three stages in the curing process (Jeffrey, 1940; Massie &
Smiley, 1974; McKee & Conrad, 1994). The first stage, the green
stage, is short in duration, lasting for 2 to 5 days. During this stage the
chlorophyll degrades, giving way to the second, the yellow stage.
Burley tobacco left in the field to wilt may continue through the green
stage while still in the field. Green color may result if leaf damage
occurs during this stage or if rapid drying conditions prevail with no
attempt to regulate the moisture loss.
The second stage is longer in duration and lasts for 5 to 10 weeks,
depending on curing conditions. The yellow pigments of the second
curing stage degrade more slowly than the chlorophyll. However,
color change is only part of the curing process during the second
stage. During the second stage many chemical reactions occur before
the leaves become dehydrated. Any factor that increases the drying
rate can leave yellow pigment in the cured leaf as well as undesirable
chemical characteristics. High ambient moisture that would delay
curing may provide fungi and bacteria with favorable conditions for
growth. Houseburn, the destruction of tobacco by these organisms,
causes loss of dry matter and reduction in quality.
The third stage of curing, the brown stage, follows as the yellow
pigment degrades and the leaf dies. Once the tobacco reaches the third
stage most of the chemical changes cease. This does not mean that the
tobacco cannot change once it reaches the brown stage. Moisture is
still influential, and tobacco may darken substantially as it takes up
and gives off moisture as the ambient humidity changes. As tobacco
takes up and gives off moisture it varies from a fragile to wet state. If
tobacco cures too quickly this process of taking up and releasing
moisture will change the color of the tobacco. However, excessive dry
conditions that halt chemical changes during curing leave unfavorable
residual products that degrade quality. Curing stopped in the green
stage leaves the green pigments in. the cured leaf. These pigments will
degrade slowly with time and favorable moisture levels to a more
desirable product.
Primed leaves cure more quickly than stalk-cured and to have more
green or yellow color than stalk-cured burley.
Temperature, humidity, air flow and sunlight control curing rate. As
temperature, air flow and sunlight increase, drying increases but
decreases as humidity increases (Jeffrey, 1940; Jeffrey, 1946).
Temperature has the greatest influence over chemical reactions during
the curing process. The desirable range is 18 to 32°C. However,
Jeffrey determined that humidity plays the more significant role due to
a narrow desirable range of 65 to 70% relative humidity (Jeffrey,
1940). In areas with little rainfall during curing, it is common to leave
tobacco exposed to sunlight on curing frames. Sunlight may
compound the negative effects of low humidity on the curing process
in these situations. Low humidity and high temperatures common
during dry curing seasons in Central America and Mexico speed
curing during the yellow stage. Variegated or 'pie-bald' tobacco
results. Quick cured tobacco is less desirable than more slowly cured
tobacco in current markets. Green or mottled tobacco is also
undesirable and is highly influenced by temperature and humidity
during the early stages of curing. Low temperature, regardless of
humidity, produces green tobacco. The degree of damage is dependent
on the extent that humidity varies from the norm (Walton & Henson,
1971). Low humidity with temperatures in the desirable range
produces greenish or mottled tobacco. As temperature increases
toward the upper limit, high humidity provides favorable conditions
for microbial activity. The resulting rot or houseburn darkens the
tobacco and reduces dry weight.
Curing Facilities
Air-cured tobacco curing facilities must take advantage of the ambient
conditions to achieve the best cure. Curing facilities vary from basic
open-air scaffolding to conventional burley barns typical of the
Bluegrass regions of Kentucky. Attributes of a good curing facility
should minimize detrimental curing conditions and protect cured
tobacco from rainfall and wind damage. Conservation of moisture and
a slower cure are desirable, especially in dry areas. Regardless of the
curing conditions, curing facilities allow little regulation of
environmental factors except for air flow. Regulation of air flow
affects temperature and humidity and controls curing. Density of
tobacco leaves or stalks in the curing structure, orientation of the
structure to

 Page 152
prevailing winds, ventilators and other openings in the structure and
the size of the structure all affect air flow. O'Bannon (1943) expressed
the relationship as:

where Q = quantity of air entering and leaving barn

V = velocity of wind
d = direction of wind, greatest when perpendicular to the
structure
A = area of ventilator opening
C = area of cracks
B = width of structure
S = spacing of tobacco
P = size of plants
As air moves through curing tobacco, moisture evaporates, so cooling
the tobacco. At the same time the ventilation increases curing. Curing
structures filled tightly will prevent proper air movement. Tightly
packed tobacco is more susceptible to houseburn. Leaving air
channels through the tobacco allows air to pass around the tobacco
and not through the tobacco, reducing the benefits of ventilation.
Spacing tobacco uniformly in both the horizontal and vertical planes
assures even curing. Tobacco curing conditions in a specific area
dictate the appropriate spacing for that area. Since air traveling across
short distances is more efficient, orientation of curing structures
broadside to the prevailing wind improves ventilation. Exceeding 12
m in width reduces the effectiveness of ventilation, while length is not
critical with proper orientation. Structures with ventilators comprising
one-third of the side surface area maximize ventilation potential.
Where air flow is minimal fans can improve ventilation. Fans
designed to pull air through the curing tobacco are more efficient than
other methods. Curing improves with appropriate regulation of the
ventilators as dictated by ambient conditions (O'Bannon, 1947;
Massie & Smiley, 1974; McKee & Conrad, 1994).
Traditional curing barns are slowly being replaced by various
temporary or semi-permanent scaffolds. These structures offer low
investment, reduced labor cost and, in some cases, improved curing
and quality. Common black plastic material serves as the roof and
siding. Although plastic provides less protection from wind and rain,
raising and lowering the plastic side curtains regulate air flow and
provide maximum conservation of moisture under dry climates. These
structures generally have less air space below the tobacco, reducing
ventilation. Keeping width to a minimum reduces the chance of rot or
houseburn. These structures do not substitute for proper curing
management. Management of ventilation is as critical as with more
conventional structures.
The conditions of cured leaves exposed to the air inside a curing
facility are good indicators of relative humidity (Table 5.11). Samples
in dry to low case or order indicate a relative humidity in the target
range of 65 to 70%. The condition of the cured leaves helps to
determine the need for more or less ventilation. If cured leaves are
placed in a protected area but exposed to ambient humidity this
further enhances the ability to assess ventilation needs properly. Cured
leaves in moderate to high case inside the curing structure indicate the
need for more ventilation. Brittle leaves signal rapid curing
conditions. Closing ventilators conserves moisture, slowing the cure.
If rapid curing conditions prevail, ventilators opened at night allow
cooler, more moist air to enter the structure. With proper regulation of
the rate of moisture loss, a good cure is possible.
Table 5.11 Feel of cured tobacco flyings in
relation to relative humidity.*
Feel of cured leaf Relative humidity (%)
High case 90 to 100
Medium to high case 85 to 90
Medium case 80 to 85
Low to medium case 75 to 80
Low case 70 to 75
Dry to low case 65 to 70
Dry 60 to 65
Dry to brittle 55 to 60
Brittle 50 to 55
Fragile 0 to 50
* From Kentucky Agricultural Experiment
Station Bulletin 501, Principles of Burley
Tobacco Barn Operation.

The use of heat to control humidity is possible, but risky and

expensive. Proper spacing and control of the heat is necessary to
prevent hot spots that set green or yellow pigments in the cured leaf.
Moderate heat produces better results, and multiple heat sources
insure distribution of heat. As heated air picks up moisture from the
tobacco, ventilation is necessary to allow moisture to escape. Natural
gas, liquid propane (LP) gas or coke are suitable fuels for curing
under high humidity. Fuels with possible impurities that could
influence quality are not suitable.

 Page 153
Market Preparation
Once tobacco completely cures, sorting or grading of tobacco can
begin. 'Fat' stems or stems that have not completely cured are an
indication that the tobacco is not ready for stripping. Fat stems can
cause rot if bulked with other cured leaves. Leaves on stalk-cut
tobacco should break clean from the stalk when fully cured. Handle
cured tobacco only when in proper case or order (moisture level).
Good lighting is important for sorting or stripping tobacco. Either
natural or fluorescent light is suitable for this process. Direct sunlight
should be avoided for best color distinction.
Grading of tobacco varies depending on the market demand. The
priming process is a form of pregrading, but further classification is
often necessary to sort out off-color variation and damaged leaves
after curing. Stalk cut tobacco must be graded as it is stripped. Leaves
are sorted by stalk position, color, leaf texture and the degree of
damage during the stripping process in stalk cut tobacco. Mixing
across stalk positions is undesirable regardless of the market. Leaf
chemical and physical handling characteristics vary considerably with
change in stalk position.
References
Anonymous (1954) Tobacco production in Kentucky. Kentucky Agric.
Ext. Serv., Lexington.
Anonymous (1993) Sulfentrazone (F6285). Technical bulletin, FMC
Corporation. Philadelphia, PA.
Anonymous (1996) 19961997 Lime and fertilizer recommendations.
Kentucky Coop. Ext. Serv., AGR-1.
Bruns, H.A. & McKee, C.G. (1985) Sucker Control Chemicals for
Maryland Tobacco. Fact Sheet 214. Cooperative Extension Service,
The University of Maryland, College Park, Maryland.
Glass, J.M. (1983) Production and leaf chemistry of burley tobacco in
Latin America. Rec. Adv. Tob. Sci., 9, 7799.
Hamilton, J.L. & Lowe, R.H. (1981) Organic matter and N effects on
soil nitrite accumulation and resultant nitrite toxicity to tobacco
transplants. Agron. J., 73, 78790.
Isaacs, S.G. (1993) Choosing a tobacco plant spacing. In: Tobacco in
Kentucky, (eds W.C. Nesmith, et al.) Kentucky Coop. Ext. Serv., ID-
73.
Jeffrey, R.N. (1940) The effect of temperature and relative humidity
during and after curing upon the quality of white burley tobacco.
Bulletin No 407. Kentucky Agricultural Experiment Station,
Lexington, Kentucky.
Jeffrey, R.N. (1946) The relation of curing conditions to quality in
burley tobacco. Bulletin No 496. Kentucky Agricultural Experiment
Station, Lexington, Kentucky.
McKee, C.G. & Conrad, D.L. (1994) Handbook on the Culture of
Maryland Tobacco, 9th edn. Maryland Tobacco Improvement
Foundation, Inc, Upper Marlboro, Maryland.
Massie, I.E. & Smiley, J.H. (1974) Harvesting and curing burley
tobacco. Agronomy Bulletin No 14. University of Kentucky,
Lexington, Kentucky.
Mulchi, C.L. (1985) Environmental factors affecting the growth,
chemistry and quality of tobacco. Rec. Adv. Tob. Sci., 11, 346.
O'Bannon, L.S. (1943) Distribution of temperature and relative
humidity within a burley tobacco barn. Bulletin No 444. Kentucky
Agricultural Experiment Station, Lexington, Kentucky.
O'Bannon, L.S. (1947) Principles of burley tobacco barn operation.
Bulletin No 501. Kentucky Agricultural Experiment Station,
Lexington, Kentucky.
Ohori, K. (1996) A Historical Review of Tobacco Production. Studies
and Technological Developments in Japan. Japan Tobacco Inc,
Tokyo.
Palmer, G.K. & Calvert, J.R. (1993) Topping, sucker, and harvest
management for burley tobacco. In: Tobacco in Kentucky, (eds W.C.
Nesmith, et al.). Kentucky Cooperative Extension Service, ID-73.
Palmer, G.K., Smiley, J.H. & Calvert, J.R. (1986) Sucker Control in
Burley and Dark Tobaccos. AGR-75. Cooperative Extension Service,
University of Kentucky, Lexington.
Pearce R.C. & Zeleznik, J. (1996) Evaluation of weed control options
for no-till burley tobacco production. In: Agronomy Research Report
1996. Progress Report 385. pp. 335. Kentucky Agricultural
Experiment Station, Lexington.
Phillips R.E. & Zeleznik, J.M. (1989) Production of burley tobacco
using no tillage and conventional tillage. J. Prod. Agr., 2, 3436.
Sims, J.L. (1980) Molybdenum Nutrition of Burley Tobacco. Agr. 82.
Department of Agronomy, University of Kentucky, Lexington.
Sims, J.L. (1993) Field selection and fertilization of tobacco. In:
Tobacco in Kentucky, (eds W.C. Nesmith, et al.) pp. 1314. Kentucky
Coop. Ext. Serv. ID-73.
Sims J.L., Thom, W.O., Wells, K.L. & Oldham, J.L. (1994) Effect of
strip band and in-row placement of phosphorus and potassium
fertilizer on growth, yield, and efficiency of potassium use by burley
tobacco. Comm. Soil Sci. Plant Anal., 25, 262738.
Thomson, W.T. (1993) Agricultural Chemicals. Book II Herbicides.
Thomson Publications, Fresno, California.
Thomson, W.T. (1995) Agricultural Chemicals. Book
III Miscellaneous Agricultural Chemicals. Thomson Publications,
Fresno, California.
Walton, L.R. & Henson, W.H., Jr (1971) Effect of environment during
curing on the quality of burley tobacco. I. Effect of low humidity
curing on support price. Tob. Sci., 15, 547.

 Page 154

5C
Oriental Tobacco
S.N. Gilchrist
R.J. Reynolds Tobacco Company
Winston-Salem, North Carolina, USA
Introduction
Opinions differ on the history and exact origin of Oriental tobaccos;
however, it is generally agreed that the varieties derived from the
main species of tobacco, Nicotiana tabacum. It is believed that seed
from native American varieties found its way to Turkey, possibly via
Spanish and English traders, and then spread, over time, to other
suitable growing areas of what was then the Ottoman Empire. This
theory would, in part, explain its extensive cultivation in the eastern
Mediterranean and Balkan regions today. In any case, cultivation of
Oriental or aromatic varieties in this part of the world is documented
as far back as the early seventeenth century. The small plant and leaf
size, characteristic of today's Oriental varieties, as well as its unique
aromatic properties are a result of the plant's adaptation to the poor
soil and stressful climatic conditions in which it developed over these
past centuries.
Commercial interest in Oriental tobacco began in the early twentieth
century when 'Turkish' cigarettes became widely popular in Europe.
At roughly the same time, the 'American Blend' was being developed
in the USA market and rapidly gained popularity. The early American
blends introduced relatively small amounts of Turkish Oriental to
smooth, soften and add aroma to the existing American-grown
Virginia and burley blends. By the end of World War II, the American
blend rapidly gained popularity in both the USA market and in Europe
and demand for Oriental tobaccos grew rapidly. Today, with the
exception of local Oriental brands in the main Oriental tobacco
producing countries, the majority of the Oriental tobacco produced is
used in blended type cigarettes produced around the world. A book
entitled, Aromatic or Oriental Tobacco (Wolfe, 1962) was used in
preparation of this monograph section.
Growing Regions and Types
Cultivation of classical Oriental tobacco today is still concentrated in
the geographical regions where it originated: near the coastal areas of
the eastern Mediterranean, Aegean, Marmara and Black Sea. Turkey,
Greece, Bulgaria and the Former Yugoslav Republic of Macedonia
(FYROM) produce the bulk of what is generally accepted as the
classical Oriental varieties. In addition to these classical types,
Oriental and semi-Oriental varieties are also grown in Italy, Albania,
Syria, Iran, Lebanon, the southern republics of the Former Soviet
Union and Thailand. The classical growing areas have a similar
climate, topography and soil conditions which have supported
Oriental tobacco cultivation for generations. In general, the ideal
climate for growing Oriental is one with a mild, wet winter and spring
and a hot, dry summer and fall. The best quality Oriental leaf is
produced in rocky, poor, somewhat infertile soil containing minimal
amounts of nitrogen and organic matter. The most suitable terrains are
generally sloping hillside fields or terraced lower mountainside
locations with good drainage and ample direct sun. As these type
fields are undesirable for most alternative crops, Oriental tobacco
production provides an invaluable cash crop in these generally poor
rural areas. The classical growing regions can be broadly divided into
three main geographical locations: Izmir, Black Sea and greater
Macedonia. While similar growing conditions as described above
exist in all three, the more subtle differences in climatic and soil
conditions between them have resulted in varietal differences that
have evolved over centuries into distinctly different types with
distinctly different physical, chemical and smoking characteristics.
With increasingly more sophisticated blend formulations, these
individual varieties within the Oriental family have been developed
into specific Oriental sub-group blends which, like flue-cured and
burley, have a specified place in total blend formulations at varying
inclusion levels in blended cigarettes.
a
Izmir Type
By far the most widely produced and used of the Oriental types is the
Izmir variety. Produced only in

 Page 155
Turkey within a radius of about 125 miles (200 km) from the Aegean
port city of Izmir (hence the name), Izmir accounts for nearly half of
the total classical Oriental production today. In recent years, annual
Izmir production has averaged around 330 million pounds (mil lb)
(150 000 metric tons (mt)), with production swings from 440 rail lb
(200 000 mt) to less than 250 rail lb (110000 mt). This considerable
elasticity of supply is less a function of demand than of political
intervention in both producer pricing and production quotas.
Izmir is a highly aromatic variety characterized by very small, bodied
leaves with a high concentration of oils. Because of the high density
of the leaf, particularly in the upper stalk positions, Izmir has rather
poor filling capacity and a slow rate of burn. Chemically it is the most
acidic of the Orientals; it is very high in sugar, 1520% (roughly
equaling that of flue-cured) and is among the lowest in nicotine,
averaging less than 1%. The highly acidic nature of Izmir is important
in blended cigarettes as it helps to smooth the smoke by neutralizing
the base characteristics of burley. Also, its slow burning properties
help to improve puff count by offsetting the more combustible burley
and Maryland types. Izmir has a distinctly aromatic smoking quality
with a sweet and sour taste and a sharp front-of-the-mouth impact.
This, coupled with the ample and consistent supply, has positioned
Izmir as the fundamental basis of the Oriental component in most
modern blended cigarettes.
b
Macedonian Basma Types
A wide variety of Basma type Oriental is produced in the area
historically known as greater Macedonia. Today this geographical
area includes northern Greece, southern Bulgaria and the
southernmost republic of FYROM. The topography and climate in this
region varies from the Aegean seaside areas of northern Greece to the
mountainous areas of south-central Bulgaria. Accordingly, the
growing conditions in specific areas of this region also differ and
while there are general similarities in the various Basma types
produced, there are also distinct differences between each variety.
Widely accepted as the finest of the Basma types, Greek Basma is a
thin, delicate variety with small leaves and a mild, pleasant aroma.
Classical Basma is produced primarily in the eastern Macedonian and
Thracian areas of northern Greece but poorer quality Basma is now
also produced, to a lesser degree, in central and western Macedonia as
well. Basma production in Greece has decreased steadily in the last
decade, largely due to the time and labor intensive nature of its
cultivation, harvest and curing. However, in recent years production
has stabilized at an average annual production level of around 55 mil
lb (25 000 mt). Greek Basma is generally considered one of the most
pleasant smoking tobaccos. It has a very fine, smooth and rich
aromatic smoking characteristic with a deep, cedary taste. Basma has
a high concentration of nicotine of from 2 to 3%, and has an average
sugar content of 10 to 12%. With increasing demand for premium
blended cigarettes, there is strong demand for Greek Basma and,
combined with the high cost of production, it commonly commands
the top price of all the Oriental varieties.
A distant relative of Basma is produced across the central and western
Macedonian regions of northern Greece. Kabakulak, meaning 'thick
ears' in Turkish, is a neutral variety that was once largely popular in
European blends but is of lesser importance in today's American blend
cigarettes. Kabakulak is generally grown in the flat plains on richer
fields, often with irrigation, and while the yields are significantly
higher than the more classical varieties, the quality and price are not
comparable. Kabakulak is a leafy, thick bodied type with little oil or
aroma. It is generally considered a neutral smoking type with little
Oriental character and is mainly used today as an Oriental filler.
Today around 35 mil lb (15000 mt) of various types are produced.
Just across the Greek border in Bulgaria, a number of Basma types are
grown. The most widely produced varieties are Djebel, Krumovgrad,
Nevrokop and Melnik, but smaller amounts of several other varieties
including Dupnitza and East Balkan are also regularly in demand.
Once a major supplier of both leaf and cigarettes to the Soviet Union
and other Eastern Bloc countries, Bulgarian production of these
Orientals and other semi-Orientals peaked at over 150 mil lb (75 000
mt). In recent years, struggling to convert to a free market system,
Bulgarian production has fallen dramatically, to just over 45 mil lb (20
000 mt). Today, with outside investment, production of the classical
varieties is slowly increasing again.
The more classical Bulgarian varieties, notably Djebel, Krumovgrad
and Nevrokop, are characteristic of a true Basma type. While
somewhat leafier and less thin and delicate, they are never-the-less
highly regarded for their rich, full, aromatic flavor and good burning
properties. They tend to be fairly low in nicotine, around 1 to 1.5%,
and have a relatively high sugar content of around 20%.

 Page 156
Due largely to a less stressful climate and more fertile growing areas,
other Bulgarian Oriental varieties tend to be more leafy and less oily
and to have a more prominent mid-rib. Some of these lesser types,
grown in low lying, plain areas, particularly in north Bulgaria, are
more semi-Oriental in nature and tend to be neutral to harsh in
smoking character.
Just west of the Bulgarian growing areas, throughout the small
country now officially known as FYROM, three main Basma types
are produced: Prilep, Yaka and Djebel. Here again, with the collapse
of the Eastern Bloc markets, production has fallen dramatically in
recent years from over 65 mil lb (30 000 mt) to around 45 mil lb (20
000 mt) in 1997.
While related, each of these three varieties has distinctive physical
and smoking characteristics that set them apart. Prilep is the most
widely produced and well known of the varieties. Grown in a wide
area around the central town of Prilep, the Prilep variety accounts for
roughly half of the country's total Oriental production. Prilep is
generally a small leaf, more bodied Basma type with a unique,
pungent aroma and a deep, cedary smoking character that is highly
regarded among blenders. The Yaka and Djebel are grown in the
south-eastern mountainous areas along the Bulgarian and Greek
borders. Similar to the classical Bulgarian varieties, they are generally
a bit more leafy and bodied and have somewhat less aroma and a
milder smoking character than Prilep. Nicotine and sugars are roughly
equivalent to their Bulgarian counterparts.
c
Samsun (Bashi Bagli) Types
'Bashi Bagli' is a Turkish term meaning 'attached at the head'.
Although not commonly used today, it was at one time used to
describe a family of Oriental varieties with heart shaped leaves and a
naked petiole by which the leaves are attached to the stalk. A number
of these varieties, including Turkish Samsun and Bafra, were grown in
north-eastern Turkey along the Black Sea and spread to parts of
southern Russia and Greece, and this descriptive term went with them.
Today it is more common to refer to the remaining varieties of this
type as Samsun types due to their origin, but the leaf shape and naked
stem are about the only remaining similarity.
The Samsun variety, as the name suggests, originated in the region
around the Black Sea port city of Samsun in north-eastern Turkey.
The land in this area rises rapidly from the sea into steeply sloping
fields on the mountain sides. Turkish Samsun grown in this area and
Bafra, a very similar variety grown just to the west around the town of
Bafra, are among the finest smoking tobaccos in the world. Over time
the distinction between the Samsun and Bafra leaf has all but
disappeared and today they are generally thought of as one and the
same and are, for the most part, bought and packed together as a
Samsun blend. Total combined production today averages around 45
rail lb (20 000 mt).
Due to the higher levels of humidity and somewhat lower
temperatures in this area, the tobaccos cure at a slower rate than in the
hot and dry Aegean regions. The result is that they take on
characteristics more like an air-cured variety, becoming darker and
more reddish than other Orientals. Samsun is sometimes referred to as
the burley of the Orient, both for its color and for its distinctive deep
resinous smoking character. The leaf tends to be of medium size, open
grained, thin and delicate and as a result has superior filling capacity
and good burning quality. Despite the comparisons to burley, Samsun
contains fairly typical Oriental nicotine and sugar levels of around 1.5
and 10%, respectively.
Greek Katerini is a prominent Samsun relative. Generally believed to
have originated from Turkish Bafra seed, Katerini has evolved over
time into a distinctly different variety with only some similarities of
leaf shape remaining. Katerini is traditionally grown in a fairly
concentrated area around the town of Katerini, just south of
Thessaloniki. The classical growing areas are hillside and lower
mountain villages that produce a small leaf, crinkly and very oily
crop. Similar to the old Bafra seed type, the leaves are heart shaped
and have a characteristic naked petiole. Unlike Samsun, Katerini has a
delicate yellow/orange, apricot color and has a pleasantly sweet,
perfume-like taste. A poorer quality Katerini is grown on the lower
plain fields which tends to be more leafy and masculine, is light and
pale in color and has a harsher smoking character. The nicotine
content of Katerini is 1.5 to 2% and sugars average around 15%. Total
production in 1997 averaged around 45 mil lb (20 000 mt).
A third Samsun type is Suchumi, originally developed in the Georgian
Republic of the Soviet Union around the Black Sea port city of
Suchum. While the original variety is now basically extinct, seed from
Russia was brought to Greece in the late 1970s where production was
first introduced in eastern Macedonia, along the Bulgarian border
around the village of Porroia. While the Greek version never quite
duplicated the fine and delicate character of the original Russian
Suchumi, a production of reasonable quality developed in this area.
The better crops produce a medium size leaf with good texture and a
strong, deep aroma. The smoking characteristics somewhat resemble
Turkish Samsun, although are more harsh

 Page 157
and strong and tend to be somewhat bitter. Later expansion of
Suchumi production into the Agrinion region of Thessaly, in southern
Greece, has been less successful. The milder climate and more fertile
fields in this area produce over-developed plants with thick, masculine
leaves and a rather prominent mid-rib and petiole. Production of
Suchum in 1997 totalled around 5000 mt.
The types and varieties described above are by no means a complete
list of the many and varied types and sub-types of Oriental and semi-
Oriental that are produced around the world. Those mentioned merely
represent the major categories and varieties commonly in demand
today. Many other growing areas and types exist and, to one extent or
another, production of each is of significance in the world tobacco
trade.
Cultivation, Harvest and Curing
As mentioned in the previous section, field selection, climate and
growing conditions are critical elements in the successful production
of quality Oriental crops. The key element in all aspects of Oriental
cultivation is plant stress. Unlike most other tobacco varieties, and
indeed most other agricultural crops, the more stressful the growing
environment and conditions are, the better the quality produced will
be. This would explain the extremely low production yields typical of
the classical Oriental varieties which average around 1125 kg/ha as
compared to 2475 kg for flue-cured and 3150 kg for burley.
The hot and dry climate of the Balkans is particularly well suited for
Oriental cultivation. Combined with the poor and rocky soil
conditions, this area is ideal for growing the stunted, small leaf plants
that characterize the classical varieties. To add further stress to plant
development, Oriental is planted with minimal space between the
plants and rows in the field, and the plants are generally not topped as
is common with most other tobacco types. Use of chemical fertilizers
and irrigation is generally not conducive to quality Oriental
production; however, their use is becoming more common as the
farmers become more modernized and commercial.
Seed Beds
Timing for the preparation of seed beds differs according to the
seasonal weather patterns in each specific growing region. In the
warmer coastal climates preparation begins in February whereas, in
the more inland areas and at higher elevations, it may not be practical
until late March. In any case, beds are not sown until after the danger
of a hard freeze has passed.
Seed bed preparation and attention is fairly similar to that of other
tobaccos types, although the number of healthy seedlings required per
hectare is obviously much higher due to the dense field populations
characteristic of Oriental cultivation. As they require considerable
attention and tending, the seed beds are generally prepared in close
proximity to the farmer's house, but may also be situated adjacent to
the prospective field in areas with an available water supply. In some
smaller villages, preparation and tending of community seed beds are
common.
The beds are generally built up above the surrounding ground level in
an area with good drainage. The soil is prepared from a mixture of
fertile dirt and sand which is sieved to assure a consistent, granular
texture. Better farmers will sterilize the soil to protect the beds from
insects and disease, using either heat or chemical sterilization
techniques. The size of the beds varies from area to area and
according to the eventual field size, but is generally 1.5 to 2 m wide
and 8 to 10 m in length. The tiny seeds are evenly sown at a rate of
approximately 5 g/m2, but a more common measure is one tiny
'Turkish' coffee cup per square meter. After the seeds are sown, a fine
layer of well fermented and sieved sheep manure is spread on top and
the bed is well watered. The beds are then covered with straw mats or
nylon sheets to protect them from frost and extreme wind and rain.
Germination takes place in about 3 weeks, and once the seedlings
have emerged the covers are removed during warm, sunny days and
replaced at night. About a week after the seedlings have emerged, a
light chemical fertilizer spray containing about 20% nitrogen is
commonly applied. If the prevailing weather is such that blue mold
and other fungi are a concern, appropriate fungicides are applied at
regular intervals. Seedling development generally takes 6 to 8 weeks
before transplanting.
Field Preparation and Cultivation
As many of the classical Oriental growing areas today are still rather
poor and primitive, small family farms are common, and much of the
work in these areas is still done by hand with crude farming
implements and beasts of burden. Mechanized farming is, however,

 Page 158
becoming more prevalent in some areas; therefore, tractors and
automatic planters are becoming more common. Unlike other tobacco
varieties, and indeed unlike most other agricultural crops, it is
common for Oriental farmers to use the same field site year after year.
This practice is largely due to the limited availability of alternative
fields for the typical small, poor Oriental growers but is in keeping
with the practice of stressing the Oriental plant. It is a general practice
to plow under the stalks and crop refuse in the fall and to over-seed
the fields with a suitable winter cover grass. The grass is planted
basically to prevent erosion, but is commonly used to graze sheep
over the winter. Springtime field preparation begins once the winter
rains have mainly subsided and the fields are dry enough to be
worked. The fields are roughly plowed to turn under the cover grass,
and a thin layer of animal manure is worked into the soil and allowed
to decompose along with the grass. In many areas it is common to
graze livestock on the fields during the winter months to keep down
the cover grasses and to add manure in the off-season. While the use
of chemical fertilizers is not recommended for quality Oriental
cultivation, they are becoming more commonly applied to increase
crop yields.
Just prior to transplanting, the fields are more thoroughly plowed and
disked to break up the soil and to smooth and level the field. Once the
seedlings are ready for transplanting, shallow furrows are prepared for
the rows of seedlings. The rows are spaced as closely as possible to
allow the minimum space required for the plant to grow and to
facilitate working in the field during the growing and harvesting
season. Depending on the eventual plant size of the particular variety,
the rows are spaced from 25 to 50 cm apart. Healthy seedlings which
have developed to about 15 cm tall are brought from the seedbeds to
the fields in baskets. Transplanting is generally done by hand in a very
time-consuming and labor-intensive process, but the use of automatic
planters pulled behind tractors is becoming more common. The
transplanting procedure involves opening two small holes per plant
very close together in a row along the side of each furrow, one hole
for the seedling and the other to apply water.
Generally, one family member walks along the furrow opening the
holes, a second follows dropping a seedling into the first hole and
firming the dirt around the roots and lower stalk and a third comes
behind with a watering can, filling each side hole with water. The
seedlings are spaced as closely together as possible to allow just
enough room for the plant to develop. The close spacing adds to the
stressful growing conditions, further stunts plant and leaf development
and improves the aromatic quality of the final product. As a general
rule of thumb, the farmers space the plants about the width of a
clenched fist apart, the rough equivalent of about 10 to 15 cm.
Applying a little arithmetic and depending on the eventual plant size
of the particular variety, the field population of Oriental tobacco crops
averages from 150 000 to 300 000 per hectare.
Due to the critical economic importance of the tobacco crop to these
poor farming families, the fields are generally very closely monitored
and cared for on a daily basis throughout the growing season. Any
sign of insect or disease damage is quickly attended to by removing
the affected plants or with the application of approved and
recommended agrochemical solutions. As tobacco is of such
importance to the overall agricultural economies of the major Oriental
producing countries, governmental agencies and/or tobacco
monopolies have well established extension services which routinely
communicate with the farming communities and regularly monitor the
crops. The regulation of growing areas and production quotas, the
monitoring and control of approved seed types and controlled use of
appropriate agrochemicals are all functions of these extension services
to varying degrees in the different producing countries.
About 3 weeks after transplanting, once the plants are well
established, the field undergoes the first cultivation. Using a short-
handled hoe, the soil around each plant is loosened and any weeds
removed. Generally, one additional cultivation is carried out within a
couple of weeks, after which the plants have filled out sufficiently to
deter further weed development. If a heavy rain should occur, an
additional light cultivation may be required to loosen the compacted
soil. For optimal quality, the plants should develop at a slow, uniform
pace from the time of transplanting through harvest. Except in
extreme heat and drought conditions, no watering or irrigation is
recommended or necessary and any further application of fertilizer is
generally detrimental to the quality.
Harvest and Curing
The lower leaves begin to ripen and yellow about 6 to 8 weeks after
transplanting, in early summer, at roughly the same time that the top
of the plant begins to flower. As mentioned earlier, Oriental varieties
are not generally topped as this practice encourages stimulation of leaf
growth and size and is detrimental to producing the

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small, compact and highly aromatic leaves desired in quality Oriental
crops. Just before harvesting commences, the better farmers will pass
through the field and strip off the lowest weak and pale ground leaves
which are of no commercial value. Harvesting is carried out in a series
of hand reapings called primings, in which only a few mature leaves
are picked at a time, as they ripen from the bottom of the stalk
upward. Historically, the plants were primed six to eight times, with
three to five ripe leaves, referred to as a hand, picked with each
priming. To reduce labor and effort, even the best farmers are making
fewer primings, and today, three to four primings of six to ten leaves
per hand are more common. Harvesting, depending upon weather
conditions, generally lasts through the summer as the crop matures
and is completed by mid-September when the very top hands are
primed. If unseasonable cool and rainy weather sets in prior to the top
hands maturing, the uppermost leaves will not ripen, and they are
either left in the fields or harvested immature and green.
Harvesting usually takes place early in the morning before the hot
summer sun descends on the fields. The primed leaves are carefully
laid, in hands, in baskets used to collect and transport them to the
curing site. The baskets of leaves harvested each day are collected in a
shady area, usually in or around the farmer's house, where the entire
family works to string the leaves onto cotton strings for curing in the
sun. This tedious task is done using long, flat needles to pierce the
midrib near the base of each leaf and then thread them onto the string.
The density of packing the leaves on the strings differs according to
the body and guminess of the particular variety and stalk position, but
care must be taken not to over-crowd the leaves which can cause
bruising and may not allow sufficient air circulation to cure the leaves
evenly. The length of the strings varies according to the variety, but is
generally uniform in length for each variety. Once each string is filled,
it is attached at either end to a bamboo pole of similar length, and the
string is tied off at intervals in the middle to distribute the weight and
prevent sagging.
In an effort to reduce the extremely time consuming and labor-
intensive process of hand stringing the leaves, a number of various
sewing machines have been developed in recent years, primarily in
Greece and Italy. While these sewing machines are commonly used
with some varieties today, the results are generally considered inferior
to traditional hand stringing. These machines sew the leaves in a flat
overlapping position which causes the tobacco to cure less evenly than
in the straight line position achieved with hand stringing. Also,
machine sewing causes more tearing and damage to the lamina than
the single hole created by hand threading, and the strings are
somewhat more difficult to remove during manipulation (processing).
Once all the leaves harvested that day are strung, the poles are hung
parallel to the ground in a shed or other cool, shady area to wilt.
Wilting generally takes from 24 to 36 hours depending on the
temperature, relative humidity and the body and degree of ripeness of
the leaves. Wilting is a critical stage of the curing process during
which the leaves gradually lose moisture and begin to yellow. If
conditions do not allow proper air circulation, heat and humidity, the
tobaccos can rot or may dry out too quickly, causing them to become
brown, brittle and lifeless. In extremely hot and dry climates, the
wilting areas are commonly sprinkled with water below and around
the racks of tobacco to help keep the temperature down and relative
humidity up during the process.
Once sufficiently wilted, the tobaccos are hung directly in the sun for
curing. There area number of various techniques for this process, each
unique to the specific variety and location. In general terms, the
strings are hung out on racks of varied construction which are situated
in such a way that they receive direct sun during the day and are
protected from harsh winds. These racks are usually built just far
enough off the ground so that the draped leaves do not touch but can
still benefit from the heat reflected off the ground or stones just below.
At night the racks are either covered with nylon sheets or the strung
leaves are moved to a covered area to protect them from the cooler
night air and morning dew. Some more bodied varieties and those
having a more prominent mid-rib require the additional curing step of
removing the strings from the rack and laying them out flat on the
ground, or on a bed of stones, to cure out the residual moisture in the
leaf and stem to prevent mold and rotting. The time required to
complete the curing process depends upon a number of variables,
including: type, stalk position, body, degree of ripeness and prevailing
weather conditions. In practice, it is the farmers' experience and
judgment that are the best indicators of proper curing.
A curing practice that is becoming more common in many areas today
employs the use of a crudely constructed hot house or curing tent
called a 'serra'. These structures are frames built over the hanging
racks described above and covered with plastic sheeting. The sides of
the serra are rolled up during the day to allow ample air circulation
while the tops remain covered to intensify the heat. At night the sides
are lowered to retain the heat and keep out the cool, damp night air.

 Page 160
Use of the serra accelerates the curing process and eliminates the
time-consuming task of covering or moving the strings each night.
While the practice is becoming inevitably more common, it is
generally considered detrimental to the curing process and the results
are inferior to the slower traditional curing method.
Once curing is completed the tobaccos are dry and very brittle and
must be handled with care to prevent breakage and loss. At this point
the tobaccos are bulked, in one form or another, so that they can be
allowed to gradually come back into a soft, pliable order during the
cool and rainy fall months. With the thinner and more delicate Basma
and Samsun types, the strings are removed from the bamboo poles and
hung from the rafters in garlands in a cool dry cellar or barn with
ample air circulation. Izmir type is generally bulked up to a meter
high in close rows, one on top of another, in a cool dry storage room.
Izmir, being a more bodied and durable type, tends to improve with
this type of bulking as the remaining moisture in the leaves migrates
uniformly throughout the stack, and some amount of preliminary
fermentation occurs under the pressure, improving both the color and
aroma.
Baling and Marketing
Late in the fall, once the tobaccos are soft and pliable enough to be
handled, the leaves are cleaned, sorted and packed into standard farm
bales and prepared for market. Here again, the size, weight and
construction of the bales differs from one variety to the next, but in
general, the tobaccos are packed and bound in such a way as to
promote safe keeping, protect the tobaccos during handling and
transport and to maintain a stable moisture level within the bale.
Simple wooden presses are used to form the bales and to assure a
consistent and uniform size and shape. The tobaccos are arranged in
the packing box so that the butt ends are facing the outside of the bale.
This allows the mid-ribs and petioles to dry more readily while
keeping the lamina inside the bale where it will stay in good condition
until market time.
There are two basic methods of baling, generally referred to by their
old Turkish names, 'dizi' and 'pastal'. Dizi refers to the baling of the
tobaccos while still on the strings on which they were hung for curing.
The strings are cut or folded to the length of the baling box and laid
head to head in rows along the bottom of the box. After carefully
laying in several layers, the tobaccos are gently pressed to compact
the bale, and then the next several layers are laid in. Once the box is
full and the desired bale size is achieved, a wooden board is laid on
top and the entire bale is compressed using a screw type hand press.
After allowing time for the tobaccos to settle and compress, the press
is released and the formed bale is removed, covered with a burlap
wrapper around three sides and firmly bound with strong hemp or
cotton cord.
The method for pastal packing is similar but involves much more
hand working of the tobaccos. The tobaccos must first be gently
removed from the strings and arranged neatly into small flat hands
(pastals) of similar stalk position and quality. This process involves
sorting out inferior, damaged or improperly cured leaves so that the
finished bales are consistent and uniform. The pastals are then laid
and stacked into the baling box and formed into bales in much the
same manner as with the dizi method. This more time consuming and
meticulous procedure is commonly used only with the finer and more
delicate Basma types, and while it is of great benefit in the eventual
grading and manipulation of the tobaccos, it also adds considerable
cost to the product. Baling of the farmer's crop is generally completed
by late fall and the bales are stored in a dry storage area until market
time. Once baling is completed in all areas for a particular variety, leaf
experts from the various merchant companies participating in the
market carry out an extensive appraisal of the crop, commonly
referred to by the Turkish term 'tespit'. This premarket grading is
conducted from farmer to farmer in the specific growing areas and
villages of interest to the prospective merchant companies. The
process is extremely time consuming and, in larger produced varieties
like Izmir with as many as 250 000 farmers, can take months to
complete. The larger merchant companies eventually grade nearly the
entire crop to plan their purchase strategy.
The various grading systems generally focus on two basic criteria,
stalk position and quality. The quality assessment takes into
consideration such key elements as ripeness, color, body, texture and
condition. Unlike most other tobacco types, Oriental farmers sell their
entire crop as a lot. That is, the buying company is obliged to offer an
average price for each farmer's entire production, from the top of the
stalk to the bottom. The ultimate aim of the premarket grading is to
establish for each farmer's crop an educated estimate of the eventual
yield of the various manipulated grades of differing commercial
values. Using this yield estimate allows the merchant company to
establish a relative value for each farmer's crop according to how it
can be used in eventual customer grades. The

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marketing season varies from country to country and by variety, but
generally runs from January through March. Unlike most other
commercial crops, it is often the case that an entire year or more has
elapsed from the time a farmer plants his crop until it is sold to the
trade and he is paid for his considerable efforts. The opening date for
the market of each variety is established by either government decree
or by consensus agreement of the various participating leaf merchants,
depending on the particular market. Market systems vary by
producing country, but generally the procedure involves the individual
farmers, circulating among the various potential buying companies,
seeking the best offer for their crop.
With the increasing importance of foreign currency financing, the
influence of exchange rates and interest rates has become a major
factor in eventual merchant cost. Accordingly, the delivery date and
payment terms have become a major point of negotiation between the
growers and merchant companies. It has become increasingly
common for merchants to offer advances or instalment payments in an
effort to delay both foreign currency borrowing and interest rate
charges. Once a price and the terms of the sale are agreed upon, a
delivery date is set. Generally, weighing dates are set up in centrally
located villages throughout the growing areas by each merchant
company, and on a specified day(s), farmers in that area transport their
crop to that location to be inspected and weighed and payment is
received. From there the tobaccos are trucked to the merchant's
warehouse to be stored until the manipulation season.
Some markets, most notably those in the European Community, have
developed a contract system whereby the merchant companies
negotiate contracts with the farmers, prior to cultivation of the crop,
for a mutually agreed quantity at a target price level according to the
eventual quality produced. While this system has stabilized
production somewhat by assuring the producers have an eventual
market for their crop, it is regularly abused by both the farmers and
merchants alike. Adjustments and refinements to the system are
regularly being studied and implemented, and perhaps this type of
system will eventually be adopted in other major producing markets.
Manipulation and Fermentation
Once the tobaccos are received in the merchant's warehouse they are
generally regraded and classified according to their suitability for the
specific blending requirements of particular customer blends. Within a
variety there are many subtle differences between different growing
regions. Many of these variations are fairly consistent from year to
year, but may also be the result of varying climatic conditions during
the growing season. While most of the major cigarette manufacturers
today prefer wide blends representing the overall characteristics of the
crop, smaller manufacturers may prefer a more specific origin or
characteristic that better suits their particular blending needs.
Depending on their customer base, a merchant will grade and classify
their purchase to achieve desired blended results.
The processing of Oriental tobaccos is commonly referred to as
manipulation. This term probably originated years ago when Oriental
leaf was opened, sorted, separated, cleaned, blended and packed,
exclusively by hand. Today manipulation is a much more modern,
mechanized process but is still very labor intensive relative to other
tobacco processing. The traditional manipulation season generally
begins in late spring, when the winter rains have subsided, and
continues through the hot summer months.
Prior to being brought to the manipulation floor the tobaccos are
conditioned. The traditional method is to lightly mist the bales with
water and cover them with damp burlap sheets for a period of time to
soften the tobaccos; however, modern conditioning units and steam
tunnels are becoming more common. The conditioned bales are
brought to the blending area which consists of multiple lines of
conveyor belts each leading to an opening machine and a series of
separators.
Along both sides of each conveyor belt are seated women who work
in pairs, removing the strings, separating the leaf pads and removing
undesirable leaves. Each line works a designated stalk position and
each pair of women works one bale at a time of that particular stalk
position and blending grade. The conveyor belt drops the padded leaf
onto a vibrating shaker with a fine mesh screen designed to remove
sand and dirt. This shaker delivers the tobacco to an incline belt which
carries it up to the infeed of the opening machine. The opening
machine consists of a series of rotating iron bars through which the
leaf pads fall. The paddling action of the rotating bars beats the
compressed pads, causing them to open and the individual leaves to
separate. The speed of the rotation and the volume of leaf input are
regulated according to the Stalk position and relative body and
condition of the tobacco being handled on each line. To avoid
excessive breakage and resulting scrap, it is crucial that the pads be
soft and

 Page 162
pliable at input. The opened leaf next passes through an air flow
separator which pulls the light, open leaves over the top and sends
them on down the line. Any heavier, unopened pads fall to the bottom
of the separator where they are pulled through a pneumatic pipe back
to the opening machine and re-run.
The opened leaf is discharged from the separator and falls onto a
series of corrugated screens which simultaneously shake and tumble
the leaf in order to sift out scrap and fines. Several workers (pickers)
are positioned along either side of these shakers to visually inspect the
tobaccos and remove any bits of string or other foreign matter, as well
as to pick out defective leaves. The scrap falling through the screens is
carried through pneumatic pipes to a scrap room where it is cleaned
and packed for sale as a byproduct. These shakers, at the end of each
blending line, feed the resulting leaf onto a single wide belt which
collects all of the various component stalk positions and grades that
will eventually be blended into a single packing grade. These leaf
components are carried to a series of large blending bulkers where
they are fed and thoroughly blended via an overhead oscillating belt.
The blended leaf is discharged alternately from each of the bulkers
and carried to automatic packing machines, usually situated one floor
below the bulkers. The packing machines are fed in a uniform manner
from the top, and the leaf is pressed into bale form by a hydraulic
press. The presses are carefully set to insure that the bales are packed
at a uniform weight and density that is appropriate for the particular
tobacco type and grade.
The packed bales are removed from the press and are kept under
uniform pressure for a period of time in order to allow the tobacco to
relax and settle enough to avoid excessive expansion during the
wrapping process. Once stabilized, the bales are removed, weighed to
assure uniformity and are placed on a wrapping table. Usually in
teams of two, workers wrap a burlap binder around three edges of the
bale and tightly lace the burlap in place using a needle and strong
hemp or cotton cord. Once tightly bound, an outer wrapper sheet of
similar burlap is inserted behind the binder strings to protect the
tobacco during handling and storage and, eventually, to be sewn in
place as the outer cover of the export bale. The finished bales are then
removed from the packing room and sent to the storage area.
Storage and Fermentation
A key element that differentiates the processing and packing of
Oriental tobacco from that of most other tobacco types is the absence
of the re-drying process. Many trials have been carried out over the
years to mechanically dry Oriental, but the process of rapidly
lowering the moisture level has repeatedly proven to adversely affect
the desirable aromatic qualities of the classical Oriental varieties.
Freshly packed Oriental tobacco may contain upwards of 20%
moisture and, particularly when modern conditioning equipment is
used during the manipulation process, great care and attention must be
paid to insure that a gradual moisture loss and fermentation occur
naturally.
The Oriental warehouse supervisor is generally considered one of the
most critical personnel in an Oriental merchant organization.
Extensive studies and research have been carried out to determine the
optimal timing and conditions for the Oriental fermentation process
but, in the end, the experience and judgment of the warehouse
supervisor are still the essential elements. The process of gradually
drying and fermenting the packed tobacco is as time-consuming and
labor-intensive as any other aspect of Oriental production and,
arguably, the most critical stage in developing and enhancing the
aromatic characteristics. A proper Oriental warehouse is dry, well
insulated and has adequate weather tight windows and doors to allow
good, uniform, cross ventilation, but that can also be sealed during
periods of adverse winds and rain. Once brought to the warehouse
from the manipulation floor, the bales are initially placed in an upright
position (kilic) on wooden racks, several tiers high. The upright
positioning of the bales, such that the leaves are oriented in a vertical
position, allows the bale to relax and spread somewhat so that the
center of the bale can breath. The bales and tiers are spaced to allow
adequate ventilation throughout the racks. The bales remain in the
kilic position throughout most of the hot summer months but are
turned and rotated, top to bottom, on a weekly basis. This traditional
laborious task (alabora) insures that each bale receives equal and
adequate ventilation during this important initial drying stage.
In late summer, once the warehouse supervisor determines that the
tobaccos have dried sufficiently, usually to around 14 to 16%, a forced
fermentation is induced. The timing of this fermentation is critical
because if the tobacco is not sufficiently dry through the center of the
bales, they may mold or over-ferment, causing a sour, sickly aroma.
On the other hand, if the tobaccos are allowed to become too dry,
proper fermentation will not occur and the desired aromatic quality
will be lost. Fermentation is traditionally induced by turning the bales
down into the position in which they are packed (istiff), i.e., such that
the leaves

 Page 163
are now in a horizontal position. The bottom tier of bales is then
compressed by the weight of two or more tiers of bales that are
stacked on top of a row of boards resting on the bales to be pressed.
The weight and pressure causes the temperature inside the bales to
increase and induces the chemical breakdown of the starches in the
leaf into sugars and acids, which is the basis of the fermentation
process. The duration of this process varies from variety to variety and
according to such other factors as the body, ripeness and condition of
the particular tobacco as well as climatic conditions. Here again, the
timing and duration of the istiff process are largely determined by the
experienced warehouse supervisor who closely monitors the tobacco
and relative conditions. Once the bottom tier is deemed ready, the
slack created in the binding cords is tightened and this tier is replaced
by the one above it, then the process is repeated.
This fermentation process, unique to Oriental tobacco, is largely
responsible for developing and intensifying the characteristic Oriental
aroma and taste. It is a complex chemical and physiological process
that is left to scientists to describe and explain in more detail. Of
greater importance to those who are responsible for selling, buying
and blending the Oriental varieties is the end result. It is the ongoing
fermentation process that, over the stored life of Oriental tobaccos,
ages and mellows the leaf, improves its characteristic color and aroma
and develops its distinctive smoking character.
Once the fermentation process is complete and cooler weather sets in,
the racks are removed and the bales are stored in stacks, flat on their
sides (plaka). In this position the remaining moisture in the bales
migrates and a uniform moisture level of 12 to 13% is maintained. At
this point, now some 20 months since the crop was planted, the
finished product is ready for sale and export. The marketing season
generally begins in late fall and exports begin toward year end.
After negotiation of the sale and inspection are completed, the bales
are covered with the outer burlap wrapper and neatly sewn for export.
The finished product is then stenciled with the buyer's required grade
marks and weighed and it is ready for shipment anywhere in the
world.
The author wishes to acknowledge the contributions of S.G. Lyon
(Socotab Leaf Tobacco Company), H.A. Bush (Socotab Leaf
Company) and R.J. English (Philip Morris, USA, retired) to this
monograph section on Oriental tobacco.
References
Wolfe, F.A. (1962) Aromatic or Oriental Tobacco. Duke University
Press, Durham, NC.

 Page 164

5D
Dark Fire-Cured Tobacco
Robert D. Miller and Donald J. Fowlkes
University of Tennessee
Knoxville, Tennessee, USA
Introduction
Dark fire-cured tobacco produced in Tennessee, Kentucky and
Virginia, USA, is used primarily for making snuff and plug chewing
tobacco. Small quantities are used in the production of specialty-type
cigars. It is characterized by a distinct aroma of wood smoke from the
open hardwood fires used during the curing process (Chaplin, et al.,
1976). Production is controlled under an acreage-based allotment
system. Allotments can be leased or transferred within the same
county, but under-or over-production cannot be carried forward to the
next year (Hourigan, 1986). There is, therefore, a strong incentive to
produce maximum yields on the allotted acreage. Type 21, dark fire-
cured tobacco, is produced in approximately 10 counties located in the
piedmont and mountain sections of Virginia (David Reed, personal
communication). Type 22 and Type 23 dark tobacco are produced in
12 counties in Tennessee (Fowlkes, et al., 1995) and 16 counties in
Kentucky (Shuffett, 1986). Type 22, which is referred to as Eastern
District fire-cured tobacco, is produced east of the Tennessee River in
the northern part of middle Tennessee and southwestern Kentucky.
Type 23 is referred to as Western District tobacco and is produced
between the Tennessee, Ohio and Mississippi Rivers in western
Kentucky and north-western Tennessee.
Although dark fire-cured tobacco is grown in relatively small regions,
it is an extremely important cash crop in these production areas. From
1991 through 1995, the average annual production of Type 22 and
Type 23 tobacco was over 16.5 million kg (Agricultural Marketing
Service, 1996). Production was split almost evenly between Kentucky
and Tennessee. The average annual value of the crop during this
period was more than $77.4 million USA. Because of the small
geographic area in which dark tobacco is produced and the low
number of purchasing companies, much of the crop is bought directly
out of the curing barns. Prices are negotiated between seller and buyer
for all or part of the crop, which is delivered directly to processing
facilities. Approximately 48% of the dark fire-cured tobacco produced
in Kentucky and Tennessee from 1991 through 1995 was sold directly
at the farm, with the remainder sold at auction warehouses
(Agricultural Marketing Service, 1996). The average price received at
the farm was $493.96 per hundred kg, compared with $446.82 at the
warehouse. The price disparity is primarily due to the fact that the
better grades of tobacco are often sold at the farm, while the less
desirable grades are marketed at warehouses.
Far less Type 21 tobacco is produced; from 1991 through 1995, the
average production was less than 1.13 million kg per year
(Agricultural Marketing Service, 1996). Sale of this tobacco produced
almost $4 million USA for Virginia tobacco producers, with an
average price of $356.92 per 100 kg. Type 21 tobacco, which is sold
at warehouses, is not as valuable as Type 22 or Type 23 because it
typically receives significantly less firing during the curing process.
The production, and particularly the curing, of dark tobacco is often
more an art than a science. Cultural practices vary significantly from
state to state, from community to community and even from neighbor
to neighbor. Many growers rely more on traditional methods, which
have been handed down from generation to generation, than they do
on University or Extension Service recommendations. Many of the
recommended practices are the same for all three states that produce
dark fire-cured tobacco, but some cultural methods differ among the
states. For example, Kentucky growers often use higher fertilization
rates and top higher (Legg & Miller, 1988) than their neighbors in
Tennessee. Tobacco produced in Virginia is fired very little in
comparison to crops produced in Kentucky and Tennessee. Plant
populations and the varieties that are grown in the three states also
vary considerably. Although the production practices that are
described herein are those generally recommended for the production
of dark fire-cured tobacco in the United States, it is impossible to
address all of the cultural variations that are used in the production of
the crop.

 Page 165
Variety Selection
One of the most important steps to successful tobacco production is
choosing the proper variety. Strong research efforts in flue-cured and
burley tobacco have resulted in considerable progress in developing
superior varieties over the years. Almost all flue-cured and burley
crops are produced from modern, disease-resistant varieties. However,
less research has been conducted on dark tobacco variety
development. Although the tobacco breeding programs at the
University of Kentucky, University of Tennessee and Virginia
Polytechnic Institute and State University have released several new
varieties in recent years, much of the dark fire-cured tobacco crop is
still produced from farmer-selected strains that have no disease
resistance. These older varieties are usually preferred because of their
superior curing characteristics. Because the success of particular
curing methods is often highly dependent on the use of a specific
variety, many growers have been reluctant to use newer, disease-
resistant varieties.
Because of the large number of individual strains, standard
descriptions of farmer-selected varieties are not possible. The most
popular varieties in Tennessee and Kentucky include Madole and
Black Mammoth. However, there are several selections of these
varieties which differ significantly (Legg & Miller, 1988). More than
a dozen strains of Madole are produced in Tennessee, with several
additional strains grown in Kentucky. In Tennessee, dark fire-cured
tobacco is often simply referred to as 'Madole tobacco'. Some of the
more popular Tennessee strains include Tom Rosson (TR), Certified,
Elliot's, Jordan's, Head's, Vincent Harris and Jennette's Madole
(Fowlkes & Miller, 1995). Some of the more popular strains in
Kentucky include Narrow Leaf (NL) Madole, Improved Madole,
Little Wood and Greenwood (Smiley, et al., 1986b). In Virginia, many
growers prefer old-line varieties such as Lizard Tail Orinoco, Hastings
or Walker's Broadleaf (Jones, 1990a).
The spread of black shank and black root rot in dark fire-cured areas
is making the use of disease-resistant varieties much more necessary
to ensure successful crop production. Type 22 and Type 23 varieties
that have black root rot resistance include KY 170 and KY 171,
developed at the University of Kentucky, and DF 911, which was
released by the University of Tennessee (Fowlkes & Miller, 1995;
Smiley, et al., 1986b). KY 171 and DF 911 are also resistant to
wildfire and tobacco mosaic virus (TMV). KY 171 is widely grown in
many areas of Kentucky. It has a more erect growth habit than most
other dark varieties. KY 171 produces about two more leaves, which
are somewhat wider and more rounded at the tips, in comparison to
older varieties. DF 911, which is produced predominantly in
Tennessee, also has an erect growth habit. It produces a large plant
that yields well. DF 911 has a larger stalk diameter than other
varieties, which makes it unpopular with many producers.
For many years DF 300 was the only dark fire-cured tobacco variety
with resistance to black shank. DF 300 was released jointly by the
Tennessee Agricultural Experiment Station and the USDA in 1968
(Latham, 1969). The variety was developed from a cross between
black shank resistant cigar variety Florida 301 and dark fire-cured
variety Broadleaf Madole. Although it has medium resistance to both
race 0 and race 1 black shank, DF 300 is susceptible to black root rot,
TMV and wildfire. Because of its low yield potential, undesirable
color and susceptibility to these important diseases, DF 300 was never
widely grown by dark tobacco producers.
DF 485, the first black shank resistant variety of dark fire-cured
tobacco that was also resistant to other tobacco diseases, was released
by the University of Tennessee in 1985 (Miller & Hunter, 1987). It has
medium resistance to race 0 and race 1 black shank and high
resistance to black root rot, wildfire and TMV. Although the yield
potential of DF 485 is high, many growers and tobacco buyers dislike
the variety. The growth habit of DF 485 is less upright and leaf color
is darker green in comparison to Madole varieties. The cured leaf of
DF 485 is darker brown in color than that of other dark fire-cured
cultivars under normal curing conditions. The quality of the cured leaf
of DF 485 is comparable to that of Madole types under normal
growing conditions. However, when it is produced in seasons that
have excessive moisture the lower leaves often have a greenish color
that results in a reduced sale price.
KY 190 was released by the Kentucky Agricultural Experiment
Station in 1989 (Legg & Tedford, 1989). KY 190, which was
developed from DF 485 and KY 171, has good resistance to both
races of black shank, black root rot, wildfire and TMV. Although it is
grown in some production areas in Kentucky, it has not been well
accepted by many Tennessee dark tobacco growers due to its
relatively low yield and quality.
Two dark fire-cured tobacco varieties have been released by the
Tennessee Agricultural Experiment Station. TN D94 (Miller, et al.,
1995) was released in 1994. It is resistant to both race 0 and race 1
black shank, black root rot, wildfire and TMV. TN D94 was

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developed by pedigree selection from a three way cross among DF
485, Certified Madole and DF 300. The growth habit of TN D94
resembles older, Madole strains of dark tobacco. Leaves have a
smooth surface similar to Madole types, rather than the 'puckered'
surface that is present in DF 485 and KY 190. This is considered
desirable in dark fire-cured tobacco as it gives the leaves a smoother,
more even finish during the curing process. TN D950 (Miller, et al.,
1998a) was released in 1995. It has high resistance to black root rot,
wildfire and TMV; it has medium resistance to race 0 and race I black
shank. Leaves have a smooth surface similar to Madole tobacco types.
In performance variety trials, growers and tobacco buyers have judged
the quality and industry acceptability of TN D950 to be equal to older,
nondisease-resistant varieties. TN D950 was compared with DF 485,
DF 300, TN D94 and Certified Madole for black shank resistance in
onfarm tests conducted in three Tennessee counties. The black shank
resistance of TN D950 was comparable to KY 190 and better than DF
485 or DF 300.
Virginia Polytechnic Institute and State University has released
several Type 21 dark fire-cured tobacco varieties (Jones, 1990a). VA
309 has medium resistance to black shank and black root rot. VA 310
and VA 331 also have medium resistance to black shank but have only
a low level of resistance to black root rot. VA 312 is susceptible to
black shank but has high resistance to black root rot and TMV.
Transplant Production
Production of an adequate supply of healthy, vigorous transplants is
essential for successful tobacco production. Excellent transplants can
be produced in conventional plant beds or by newer, hydroponic
production methods. When producing transplants in conventional
plant beds, growers must pay attention to key factors such as site,
fumigation, fertilization, seeding date and rate, cover management,
water management and insect and disease control to consistently
produce desirable transplants. Enough plant bed area should be
prepared to provide adequate plants for transplanting the entire crop in
two pullings. For dark tobacco, 41.8 to 55.7 m2 of bed space should
be adequate to transplant about one half of a hectare (one acre)
(Smiley, et al., 1986a; Fowlkes, 1989).
Careful consideration should be given to the plant bed site. The best
sites for plant beds have a convenient source of water, preferably from
a well or utility line. Soils should be fertile, well drained and high in
organic matter. The site should be free from shade, gently sloped to
the south or south-east and contain windbreaks on the north and north-
west sides (Fowlkes, 1989). Sites near tobacco barns and areas
infested with tobacco trash, bullnettles, ground cherries or plantains
should be avoided since they may be sources of tobacco diseases
(Smiley, et al., 1986a). Areas heavily infested with white clover or
dodder should also be avoided since these weeds are difficult to
control in plant beds.
Weed control is critical to the production of an adequate supply of
healthy plants. Most weeds can be controlled by fumigating the beds
with methyl bromide prior to seeding. Although beds can be
fumigated during the previous fall or in the spring that the crop is to
be produced, fumigation during September or October is
recommended because soil temperatures are higher and wet weather is
less likely to occur. Timely spring treatments are often difficult due to
cold, wet soils. Soils should be moist and above 10°C for effective
fumigation. The soil should be thoroughly worked just prior to
fumigation. If methyl bromide is used, it is applied at the rate of 454 g
per 9.3 m2 (Smiley, et al., 1986a; Fowlkes, 1989). It can be applied as
a liquid in pressurized 454 g metal cans. When the cans are punctured
in the applicator tray, gas is formed. The gas, held in contact with the
soil by a plastic cover, kills weed seed as it penetrates the upper 10.2
to 15.2 cm of loose soil. Methyl bromide can be applied by
mechanical equipment which injects the gas into the soil and covers
the bed in one operation. The plastic cover should be left on the bed
for at least 48 hours before it is removed. When beds are fumigated in
the fall, the covers are typically left in place until spring to prevent
weed seed from blowing onto the bed.
The soil can also be fumigated using Vapam (metam), a liquid
material. For best results Vapam should be used in the fall (Fowlkes,
1989). Spring treatments are not recommended. Vapam is applied at
the rate of 3.79 to 7.56 L of chemical in a minimum of 151 L of water
per 83.6 m2 of bed. A sprinkler can, a sprayer with a high volume
handgun attachment or other equipment can be used to uniformly treat
the bed. Immediately after treatment, the bed is covered with a plastic
tarp, which is left in place over the winter.
Seeding the plant beds can be done anytime that the weather permits,
but usually occurs in late February or early March. After removal of
the plastic covers, a complete analysis fertilizer, such as 4-16-4 or 10-
10-10, is applied to the soil at rates to provide 1.81 to 2.27 kg of
nitrogen per 83.6 m2 of bed. The soil is raked lightly, but no deeper
than 5.08 cm, to incorporate the fertilizer and to help aerate the soil.
The bed is then seeded at

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the rate of 4.72 g (two level teaspoons) of seed per 83.6 m2 (Smiley,
et al., 1986a; Fowlkes, 1989). A thin layer of fumigated straw mulch
is applied to protect the seedlings and the bed is covered with a cotton,
polypropylene or nylon cover. The covers are left on the beds until
approximately 1 week before transplanting, when they are removed to
harden the transplants.
Lack of moisture during the seed germination period is the major
cause of plant stand problems. Beds should be watered immediately
after seeding, and as needed throughout the transplant production
period. Thorough watering every 4 to 5 days is better than applying
small amounts every day or two (Smiley, et al., 1986a). Application of
2.54 cm of irrigation to a 83.6 m2 bed requires about 2120 L of water.
Well or utility water should be used, since water from streams and
ponds may contain disease organisms. Beds should be watered during
the morning hours rather than during late afternoon or at night to
reduce the incidence of diseases.
The beds should be monitored closely for the development of diseases
or presence of insect pests. They should be sprayed every 7 to 10 days
with Dithane 11F (mancozeb) or other labeled fungicides to prevent
the occurrence of fungal diseases such as blue mold or anthracnose,
and with streptomycin to prevent the development of angular leaf
spot. Labeled insecticides should be applied to control insect pests.
The use of hydroponic tobacco transplants, commonly referred to as
'float plants', is fast gaining in popularity among tobacco producers.
This system, in which plants are grown in styrofoam trays filled with
soil-less potting mix that are floated on water in 'float beds' or 'water
beds', has several advantages over traditional bed plants. These
advantages include:
(1) elimination of plant pulling;
(2) reduced risk of plant bed failures due to insufficient water or poor
weed control;
(3) easier management of plant growth during wet transplanting
seasons;
(4) the ability to save unused plants resulting from unexpected rainfall
by re-floating trays on the water beds;
(5) increased daily transplanting rates since plant pulling is eliminated
and transplanting can be done during any time of the day; and
(6) better plant survival and early season growth of float plants in
comparison to conventional transplants (Fowlkes, 1997; Miller, et al.,
1998b).
Growers have five options in using tobacco float plants:
(1) purchase plants that are ready to transplant from commercial
greenhouse operators;
(2) buy prestarted plants from greenhouse operators and finish to
transplant size in float beds;
(3) buy prestarted plugs, which are transferred into trays with larger
cells and finished in float beds;
(4) start seedlings in small plastic trays and later transfer them into
float trays; or
(5) direct seed and grow their own plants (Fowlkes, 1997).
The use of hydroponic tobacco transplants began in the late 1980s and
has increased rapidly each year. It is estimated that over 60% of the
1996 crop was produced using hydroponic plants (Fowlkes, 1997).
Although many growers initially utilized the transfer of prestarted
plugs or seedlings into float trays which they then finished in their
own float beds, most producers are now either purchasing finished
transplants or direct seeding and producing their own plants.
Direct seeding involves growing the plants from seed to transplant
size in the float trays. Seedlings are germinated in the same tray from
which they will be transplanted, thereby eliminating the task of
transferring seedlings from starter trays to larger celled trays used for
finishing the plants. This reduces the labor requirement since seeding
a tray is much faster than transferring seedlings into a tray. However,
it significantly increases the management required because care must
be taken to provide a favorable environment for seedling germination
and early growth. The direct seeding method of producing tobacco
float plants has been primarily used by larger producers who have
purchased greenhouses.
Maintaining a favorable environment is crucial to the successful
hydroponic production of tobacco transplants (Duncan & Anderson,
1997; Fowlkes, 1997; Pearce & Maksymowicz, 1997). Because each
cell in the styrofoam trays receives only one seed, a high, uniform
germination rate is essential. Many growers have chosen to purchase
environmentally controlled greenhouses for transplant production.
They typically have vented heating systems and exhaust and
circulation fans to regulate temperature and humidity. Optimum seed
germination occurs when the average air temperature is maintained
between 18.3 and 23.9°C. Germination will occur within 5 to 10 days
when temperatures are maintained within this range. Excessive
temperatures are much more detrimental to seed germination than
insufficient heat. Cool temperatures will delay seed germination, but
temperatures above 32.2°C will deactivate enzymes and result

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in decreased seed germination (Fowlkes, 1997; Miller, et al., 1998b).
Humidity control is important, particularly in preventing or
minimizing diseases in developing plants.
Ventilation is best achieved with large exhaust fanlouver systems
(Duncan & Anderson, 1997; Fowlkes, 1997). The ventilation system
should be able to provide one air exchange per minute for the entire
greenhouse. Smaller fans are usually mounted inside the greenhouse
to circulate air, which helps to equalize temperatures and humidity
throughout the house. Some greenhouses are also equipped with side
curtains to further facilitate ventilation. The fans and curtains may be
manually or thermostatically controlled.
The number of trays needed per hectare of tobacco will depend upon
the number of plants set per hectare, the number of cells per tray, and
the percent usable plants produced per tray. Although all tobacco float
trays are approximately 68.6 cm × 35.6 cm in size, the number of cells
per tray varies. A University of Tennessee study found no significant
difference in transplant survival or percent usable plants among plants
from trays with 200, 242, 253, 288, 338 and 392 cells (Fowlkes,
1997). Because the number of plants produced per unit area
significantly affects the cost of transplant production in expensive
greenhouses, 288, 338 or 392 cell trays are typically used to maximize
the number of plants that can be produced per square meter. The
percentage of usable plants will depend upon how closely the plants
are managed. With direct seeding, a figure of 75 to 85% is typical
when the plants are not clipped. Clipping two to three times will often
improve percent usability to 8590%. Many greenhouse owners will
clip plants four to six times to increase uniformity of plants; this often
results in greater than 90% plant usability. The higher cell number
trays will produce a smaller sized plant that must be clipped more
often to produce a comparable sized plant, especially in terms of stem
diameter. Disease control may require more attention in trays with a
greater density of plants and therefore restricted air circulation.
However, with proper management these trays will produce
acceptable transplants.
Hydroponic transplants are produced in soil-less potting mixes
specifically formulated for tobacco float systems. The trays are filled
with mix and 'dibbled' prior to seeding. This involves pressing the
trays with a mold in order to make holes in the mix. Dibbling gently
compresses the potting mix, centers the seed in the cell and helps
create a favorable germination environment. Dibble boards typically
consist of either pyramid or round shaped objects (forged metal,
marble, etc.) attached to a metal or wood base with handles on both
ends. Although shallow dibbling is typically used in most greenhouse
operations where temperature and humidity are closely controlled,
dibble depths of 1.3 cm or deeper result in improved germination in
outdoor beds (Miller, et al., 1998b).
After the trays have been dibbled, they are ready for seeding. Because
natural tobacco seed are too small to allow mechanical placement of
individual seed, they must be pelletized with clay to increase their
size. Both primed and nonprimed pellefized seed of most popular
tobacco varieties are readily available from commercial seed
companies. Priming refers to a process where germination is begun,
then suspended, before the seed is pelletized. Primed seed typically
germinate quicker and at lower temperatures than nonprimed seed
(Millet, et al., 1998b). Mechanical seeders are used to rapidly place
one seed into every tray cell. Commercial seeders and dibble boards
are used by many growers. These seeders are usually vacuum
operated and range in cost from a few hundred to over three thousand
dollars. As an alternative, some growers choose to build or buy low-
cost hand made seeders and dibble boards (Miller, et al., 1994). The
seed should not be covered after seeding. Trays should be handled
gently after seeding to prevent potting mix from covering the seed.
Covered seed will germinate poorly if at all.
Fortified potting mixes with time released fertilizer are available, but
it is difficult to regulate plant growth with these materials due to the
uncertainty of remaining nutrient levels as the seedlings approach
transplanting size. It is easier to regulate plant growth by using
nonfertilized potting mix and adding necessary nutrients to the water.
A water-soluble fertilizer such as 20-10-20 or 20-20-20 is typically
applied to the float bed water. The fertilizer should have nitrogen in
the nitrate form rather than urea and contain micronutrients in addition
to the N-P-K. Nitrogen rates should be maintained in the range of 75
to 125 µg/g to provide good plant growth and development (Fowlkes,
1997; Pearce & Maksymowicz, 1997). This requires around 142 to
227 g of 20-10-20 or 20-20-20 fertilizer per 379 L of float bed water.
One advantage of float plants is that growth can be regulated as they
approach transplant size. Studies have shown that plants grown at a
steady, moderate rate and transplanted as soon as they reach adequate
size usually perform best (Fowlkes, 1997). However, it may be
necessary to temporarily hold plants if weather conditions are
unsuitable for transplanting. The growth of plants can be suspended
by simply replacing the fertilized water in the beds with nonfertilized
water

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and clipping as necessary. Plants will continue to grow, however, until
all of the nutrients that are in the root pod are depleted. This typically
takes 5 to 8 days if the nitrogen level is in the range of 75 to 125 µg/g
before the water is replaced (Miller, et al., 1998b). Plants can be held
for 3 to 5 weeks if necessary. However, they should be transplanted as
soon as possible for optimum performance. Older plants will yellow
and essentially go dormant. Once they are eventually transplanted,
they are slower to start growing than plants that are transplanted as
soon as they are large enough.
Research conducted at the University of Tennessee Agricultural
Experiment Station has demonstrated that the production of
transplants by direct seeding in outdoor float beds is a safe, reliable
method if proper management procedures are followed (Miller, et al.,
1998b). Many smaller burley and dark fire-cured tobacco producers
are now using this system. Float beds are made with a treated lumber
frame (usually 5.1 cm × 15.2 cm or 5.1 cm × 20.3 cm) and 6 mil (151
micron) black plastic lining. The bed site must be prepared so that it is
level and smooth. Sand or similar material is used to make a soft
foundation for the bed liner. Stakes or soil placed around the outside
perimeter of the bed prevent the sides of the frame from bowing
outward when water is added. Although the bed frame can be built to
hold any number of trays, a 6.1 m × 2.1 m bed is recommended. Most
float bed covers come in 15.2 m lengths, which is enough to cover
two 6.1 m beds without having wasted material. It is more difficult to
prevent damage from water drips and to maintain desired
temperatures in larger beds, and losses from diseases or water leaks
can be minimized by limiting the size of beds. A 6.1 m × 2.1 m bed
will hold 51 trays, which will produce enough plants for
approximately 4.9 ha of dark fire-cured tobacco. Growers who direct
seed in outdoor beds are less concerned about the number of plants
per square meter and usually prefer trays having 200, 242, 253 or 288
cells.
It is very important to protect outside beds from water damage which
can result from heavy rainfall or condensation (Fowlkes, 1997; Miller,
et al., 1998b; Pearce & Maksymowicz, 1997). If water drips onto
newly seeded trays, the seed may be buried and fail to emerge. The
beds are covered with a heavy spunbonded polypropylene material
that is specially designed for use with outdoor float beds and will shed
rainfall unless it is extremely heavy. The bed covers are supported
above the plants by 1.9 cm PVC or metal bows. Although the support
bows are often attached directly to the bed frame, it is preferable to
construct a light-weight cap for the beds out of 5 cm × 10 cm studs
(Miller, et al., 1998b). The cap can be easily removed for inspection
of plants and application of pesticides; it also makes it much easier to
regulate temperatures as the weather begins to get warmer in late
spring. Arched PVC bows are easily constructed by connecting two
25 cm uprights to 230 cm cross-pieces using 45° unions. Although the
bows may be arched or A-frame in shape, it is extremely important
that they be constructed with adequate height and pitch to effectively
shed rainfall. A bow height of about 50.8 cm above the plants is
sufficient for a bed that is 2.1 m wide; wider beds require higher
arches to shed water effectively.
Maintaining proper temperatures inside the bed is essential for
successful production of tobacco transplants. Several studies have
been conducted at the University of Tennessee Tobacco Experiment
Station (TES) to determine the effects of temperature on germination
of tobacco seed in outdoor float beds (Miller, et al., 1998b).
Thermocouples were placed at various locations in the beds to
monitor air and water temperatures every 30 minutes from seeding to
transplanting. Although the optimum temperature for germination is
approximately 21.1°C, data from the Tennessee studies demonstrated
that tobacco seed will germinate at significantly lower temperatures.
However, there is a direct relationship between temperature and the
rate and uniformity of seed germination. The average air temperature
inside the bed must be approximately 18.3°C in order to obtain
complete germination of primed seed within 7 days. Lower
temperatures resulted in delayed seed germination. If the average
temperature inside the bed was 15.6°C, 10 days were required for
germination of primed seed; 14 days were required if the temperature
was as low as 10.0°C. Nonprimed seed did not consistently germinate
within 7 days, regardless of temperature. An average temperature of
18.3°C inside the bed resulted in germination of nonprimed seed
within 10 days, while a temperature of 12.8°C resulted in germination
within 14 days.
If outdoor float beds are seeded from mid-March to early April in dark
fire-cured production areas, supplemental heat is usually required to
maintain adequate temperatures inside the beds for satisfactory seed
germination. The amount of supplemental heat required is determined
by the average ambient air temperature that occurs within the first 5 to
7 days after seeding. The TES research studies demonstrated that the
most effective method of maintaining desired air temperatures inside
the bed is to heat the water to

 Page 170
26.7°C by using two thermostatically controlled electric water bed
heaters for each 6.1 m × 2.1 m bed (Miller, et al., 1998b). The heaters
are rubber mats that are used to warm the water in regular water bed
mattresses. The beds should be covered with 6 mil (151 micron) black
plastic to minimize heat loss when temperatures are predicted to be
below 4.4°C. However, they should never be covered with clear
plastic. Temperatures inside beds covered with clear plastic can
exceed 48.9°C very quickly when the sun is shining, resulting in
extreme injury or death of the plants. The heaters can be reduced to
about 15.6 to 18.3°C after germination is complete, and turned off
when frost damage is no longer a threat.
This method of supplying supplemental heat has been shown to be
very effective in preventing cold damage in float beds. In the TES
studies evaluating this system, the average temperature inside float
beds was maintained at 18.3°C, which is adequate for seed
germination, when the average outside air temperature was 10.0°C
(Miller, et al., 1998b). Even when the outside temperature dropped to
-10.6°C, the temperature inside the heated beds was never lower than
3.3°C if they were covered with 6 mil (151 micron) black plastic. An
extremely low temperature of -14.4°C caused temperatures inside the
bed to drop to -1.1°C, but little damage to tobacco seedlings was
observed. This suggests that the risk of freezing temperatures
occurring inside the bed is minimal with this system. Although the
amount of time required for the seedlings to reach transplant size will
vary somewhat with prevailing weather conditions, plants in heated
beds were ready within 52 to 60 days after seeding in 3 consecutive
years of research at TES.
Although supplemental heat greatly increases the reliability of
transplant production in outdoor float beds, it is not always necessary
(Miller, et al., 1998b). Plants can be successfully grown without heat
if temperatures below -2.2° to -1.1°C are not encountered. For
optimum results, beds should not be seeded until the average 24 hour
temperature exceeds 12.8°C. Seed germination is much slower
without supplemental heat, and an additional 10 to 17 days may be
required for seedlings to reach transplant size, but excellent results
can be achieved during warm spring seasons. However, production of
tobacco plants by direct seeding without supplemental heat is
extremely risky.
Many growers attempt to avoid using supplemental heat by delaying
seeding until mid-April, but it is very difficult to prevent the beds
from overheating on warm, sunny days. In the research conducted at
TES, percent germination decreased substantially when temperatures
inside the beds exceeded 32.2°C (Miller, et al., 1998b). This was
particularly true during the first week after seeding. Excessive
temperatures occurred on bright, sunny days when outdoor
temperatures were 21.1°C or higher; temperatures inside the beds
always exceeded 32.2°C when ambient temperatures were above
26.7°C. Excessively high temperatures were prevented by raising the
covers by 15.2 to 25.4 cm to ventilate the beds. This was most easily
accomplished by using removable 'caps' to cover the beds rather than
attaching the cover material directly to the bed; spacers were simply
placed between the beds and the caps to provide ventilation.
Although clipping is a standard practice in most greenhouse
operations, it is an optional practice with direct seeding. In a 2-year
study conducted at TES, the percentage of usable plants ranged from
81 to 89% for nonclipped plants and 77 to 92% for plants that were
clipped twice (Miller, et al., 1998b). The average transplant usability
rate was approximately 5% higher for clipped plants in comparison to
nonclipped plants. Because it is much cheaper to produce tobacco
transplants in outdoor beds than it is in greenhouses, many growers
choose not to clip. However, clipping may be necessary to increase
plant uniformity, particularly in nonheated beds, if germination is
extended over several days.
Site Selection and Fertilization
Site selection for the production of dark fire-cured tobacco is very
important because of the limited number of disease resistant varieties
that are available. Deep, well drained soils high in organic matter are
most desirable; in general, the most productive land on the farm is the
most profitable for dark tobacco production (Everette, 1958; Rhodes,
et al., 1980; Jones, 1990a). Because of the prevalence of black shank
and black root rot in most production areas and the lack of disease
resistance in many older dark tobacco varieties, crop rotation is
essential. Careful consideration should be given to weed problems
when selecting fields for tobacco production. Carryover from
persistent herbicides used in other crops in the rotation, such as
atrazine in corn, that tobacco is sensitive to, should be considered. A
3-year rotation of tobacco with grass crops such as small grains,
fescue or orchardgrass is usually recommended (Jones, 1990a;
Maksymowicz, 1994; Fowlkes, et al., 1995). Legumes play an
important role in rotations for burley and flue-cured tobacco, but are
not used extensively in dark tobacco because of

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problems that can occur from black root rot when nonresistant
cultivars are used (Fowlkes, et al., 1995). Red clover can be beneficial
in the rotation, but only black root rot resistant tobacco varieties
should be grown (Johnson, 1990a). Cover crops are normally turned
under at least 3 weeks prior to transplanting to facilitate good soil
preparation.
Maintaining proper soil pH is critical for high yields of quality dark
tobacco. Although soil pH levels between 5.6 and 6.5 are often
recommended for burley and flue-cured tobacco, lower levels are
usually considered desirable for dark tobacco (Everette, 1958;
Rhodes, et al., 1980; Jones, 1990a). If the pH exceeds 6.0, black root
rot may be more prevalent when nonresistant varieties are used.
Studies conducted in Tennessee in the 1960s indicated that soils with
a pH range from 5.4 to 5.8 appeared to be most desirable for dark
tobacco production (Parks & Safley, 1967). However, more recent
studies suggest that pH ranges from 5.5 to 6.0 are more desirable
(Jones, 1990a; Fowlkes, et al., 1995). Toxicity resulting from high
concentrations of soil manganese is one of the most prevalent
disorders in tobacco production. If the soil pH falls much below 5.6,
excessive levels of manganese are released into the soil (Fowlkes, et
al., 1995). Manganese toxicity in tobacco results in slow early season
growth, often delaying maturity by 1 to 3 weeks. Pale green or
yellowish coloring develops between the larger veins of leaves on
small plants, with the tips of lower leaves being the first to show
symptoms. In severe cases, numerous dead spots develop on the
leaves and plants are severely stunted, resulting in substantial
reductions in the yield and quality of the cured leaves. The best way to
prevent manganese toxicity in tobacco is to maintain an adequate soil
pH by adding appropriate amounts of ground limestone.
Dark fire-cured tobacco requires relatively large amounts of fertilizer
to produce high yields of good quality leaf. Nitrogen, phosphorous
and potassium are the most important nutrients for tobacco
production. Annual soil testing is essential to ensure that proper
amounts of fertilizer are applied to the crop. The amount of nitrogen
recommended for dark fire-cured tobacco varies among the three
states that produce the crop. In Tennessee, it is recommended that
total nitrogen from all sources should not exceed 225 kg per ha
(Fowlkes, et al., 1995). Levels as high as 281 to 338 kg/ha are
recommended in Kentucky (Pearce, 1997). Much less nitrogen is used
in Type 21 tobacco, with current Virginia recommendations
suggesting only 141 to 169 kg/ha (Jones, 1990a). Excessive nitrogen
results in rank growth, delayed maturity, decreased quality, reduced
soil pH and increased weed pressure (Fowlkes, et al., 1995). All of the
nitrogen can be applied along with the phosphorous and potassium
and disked into the soil prior to transplanting, or part can be applied
by side dressing 2 to 3 weeks after transplanting (Jones, 1990a;
Fowlkes, et al., 1995). All commonly available sources of nitrogen
can be used successfully for dark tobacco if proper soil pH is
maintained. However, nonacid-forming sources of nitrogen, such as
sodium nitrate or calcium nitrate, will help prevent damaging
decreases in soil pH. If a leguminous cover crop or manure is turned
under during soil preparation, less inorganic fertilizer is needed
(Jones, 1990a). These practices are normally avoided in dark tobacco
production, however, unless a black root rot resistant variety is
planted or good crop rotation is practised.
Dark tobacco requires adequate amounts of phosphate for early
growth and proper maturity (Jones, 1990a; Fowlkes, et al., 1995).
Many soils used for tobacco production have relatively high levels of
phosphate due to repeated fertilization over several years. Additional
phosphate requirements may range from 67.5 kg/ha for high testing
soils to 169 kg/ha for soils testing low in phosphate. Potassium levels
for dark tobacco production are relatively high; a high potassium
content is necessary for good cured leaf content (Jones, 1990a).
Recommendations range from 135 to 338 kg/ha, depending on levels
already present in the soil. Since high levels of chloride in tobacco can
result in poor curing and undesirable leaf quality, sulfate of potash,
rather than the muriate form, is recommended.
Weed Control
Growers should consider existing weed problems when selecting a
field for tobacco production. Many weeds such as ground cherry,
jimson weed, horsenettle, cocklebur, bermudagrass and rhizome
johnsongrass are not adequately controlled by existing tobacco
herbicides (Rhodes & Fowlkes, 1995). A combination of soilapplied
herbicides and timely, shallow cultivation provides adequate weed
control in most situations. Recommended herbicides for dark tobacco
include Balan (benfluralin), Devrinol (napropamide), Prowl
(pendimethalin), Tillam (pebulate) and Spartan (sulfentrazone).
Proper application and incorporation and adequate soil moisture are
critical for effective weed control.
Depending on the herbicide, application may be prior to transplanting,
over the top after transplanting or at lay-by. With the exception of
Spartan, all of the

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herbicides labeled for tobacco may be incorporated prior to
transplanting. Spartan is applied prior to transplanting, but is not
incorporated into the soil. Advantages of pretransplant incorporation
of herbicides include the ability to tank-mix with other pesticides to
reduce application costs, reduced dependence on rainfall or irrigation
for incorporation and activation and more consistent weed control due
to better soil incorporation. The primary disadvantage of pretransplant
incorporation is early season root pruning which can delay initial
growth of the plants. This can usually be eliminated by avoiding
higher than recommended application rates and by proper
incorporation into the soil. Sprayers need to be carefully calibrated
and spray overlap should be avoided. Chemicals should only be
incorporated in the top 2.5 cm to 5 cm of the soil. Deeper
incorporation will result in damage to transplant roots and will give
erratic weed control. Herbicides can be adequately incorporated by
setting a disk to cut 10 cm to 15 cm deep; this will incorporate the
chemical into the top 5 cm of soil. The soil should be disked twice,
with the second pass preferably made at a right angle to the first
(Johnson, 1990b).
Devrinol 50DF is presently the only tobacco herbicide labeled for
application over the top after transplanting (Johnson, 1990b; Rhodes
& Fowlkes, 1995). It must be applied within 7 days following
transplanting and requires immediate tillage. Application in this
manner has several advantages if native weed infestations are fairly
low. It saves a trip across the field, which may be particularly
beneficial during wet seasons when field access is limited. Herbicides
are only applied in a 40 cm to 70 cm band over the row, which
requires less material, and the possibility of early season root pruning
is minimized. However, proper activation of the herbicide is
dependent upon approximately 2.54 cm of precipitation or irrigation
within 2 to 3 days after application. Inadequate weed control often
results because of insufficient soil moisture, particularly in heavily
infested fields.
Devrinol 50DF and Prowl 4E are also labeled for lay-by application
(Johnson, 1990b; Rhodes & Fowlkes, 1995). Herbicides are directed
toward the middle of the rows during the last cultivation of the crop to
extend weed control throughout the season. If Prowl 4E is used, care
should be taken to prevent the herbicide from contacting the plants.
Lay-by applications are particularly useful in cases where inadequate
weed control was obtained from pre-plant or over the top herbicide
applications, additional weed seed in untreated soil were brought to
the surface by deep cultivation or a herbicide with a short residual was
used prior to transplanting. Rainfall or irrigation of 2.54 cm is
required within 2 to 3 days to ensure herbicide activation.
Transplanting and Plant Spacing
Transplanting of dark tobacco can be done anytime the plants are
ready and weather conditions are favorable. Good transplanting
conditions exist in dark tobacco production areas in the USA from 10
May through 1 June during most years. The threat of frost is gone and
yet the weather has not gotten hot enough to cause problems with
stand establishment. Stand reductions from hot, dry weather
commonly occur in tobacco transplanted after the middle of June.
Because float plants have a much more extensive root system than
plant bed plants, they undergo less transplant shock and can be
transplanted more successfully under unfavorable conditions
(Fowlkes, et al., 1995). Transplant shock in conventional plant bed
plants can be reduced by watering the bed before plant pulling. The
use of healthy, vigorous plants that are kept in the shade with roots
moist until transplanting will greatly improve plant survival. If plants
must be stored temporarily, they should be kept in a cool place with
the leaves dry. At least 1870, and preferably 28053740, liters of water
should be used per hectare when transplanting. Transplanting during
mid-to-late afternoon or early evening will minimize sun and heat
damage on hot, sunny days. Transplanting should be avoided if
possible when temperatures exceed 30°C.
Studies show that float plants survive transplanting better and start
growing more rapidly than traditional bed plants (Fowlkes, et al.,
1995; Fowlkes, 1997). Float plants can be transplanted as soon as the
root ball fills the cell and the plants are large enough to be held by the
fingers of a conventional transplanter. Smaller float transplants
generally perform better than larger ones. Float plants can be set
throughout the day with little or no transplant shock unless they have
been over-fertilized and are extremely tender and succulent. Plants
should be placed well into the ground, making sure the entire root ball
is underground. This minimizes the possibility of plants leaning or
partially falling over, which can cause early season ground suckering
and increase lodging incidence later in the season.
In Kentucky and Tennessee, planting densities may vary from
approximately 9884 to 12849 plants per ha (Fowlkes, et al., 1995;
Maksymowicz, 1994). Dark tobacco plants are typically spaced 102 to

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112 cm between rows and 76 to 91 cm between plants within the row
in these states. Plant populations of between 12355 and 13220 per ha
are typically recommended in Virginia, with a row spacing of 107 cm
and plants spaced 71 to 76 cm within the row (Jones, 1990a). A 107
cm row and 91 cm plant spacing combination has been popular among
Tennessee growers attempting to produce high quality, wrapper grade
tobacco. This spacing provides approximately 10255 plants per ha.
Research conducted from 1988 to 1990 at the Highland Rim
Experiment Station in Springfield, Tennessee, investigated the effects
of row width and plant spacing on three dark fire-cured tobacco
varieties (Fowlkes, et al., 1995). Plant spacings of 96.5 or 106.7 cm
between rows and 70.0, 76.2 or 91.4 cm between plants within a row
were evaluated. Narrowing the row spacing from 106.7 to 96.5 cm
reduced yield and quality, regardless of within row plant spacing. This
was thought to be a result of physical damage to the leaves that
occurred during mechanized cultivation and spraying, although
reduced light penetration could not be ruled out. Closer spacing of
plants within the rows resulted in significant increases in yields. A
spacing of 70.0 cm within the row averaged a 534 kg/ha higher yield
than a 91.4 cm spacing. The highest yield was obtained with 70.0 cm
plant spacings in 106.7 cm rows. In this study, within row spacing had
no effect on cured leaf quality as measured by grade index (Miller &
Legg, 1990). However, decreased within row plant spacing generally
resulted in smaller leaves. More significantly, higher plant populations
resulted in thinner-bodied leaves and a lower percentage of wrapper
grade tobacco. Although the three cultivars used in the study were
significantly different for leaf size and growth habit, no significant
variety by plant spacing interactions were detected.
Disease Control
Plant diseases cause significant yield reductions in dark tobacco each
year. Because the number of dark tobacco varieties with disease
resistance is relatively limited, adequate disease control is dependent
on proper management practices. Good disease control requires the
use of several, integrated control practices. Careful consideration
should be given to disease problems occurring in each field. Proper
crop rotation and the use of resistant varieties and pesticides will
usually ensure the successful production of a dark fire-cured tobacco
crop.
The two most important diseases of dark tobacco are black shank and
black root rot (Fowlkes, et al., 1995). Black shank is caused by the
soil-borne fungus Phytophora parasitica (Lucas, 1975). Two strains
of the fungus (race 0 and race 1) occur in tobacco producing areas of
the United States. Research conducted in 1981 (Hunter, et al., 1981)
demonstrated that both races were present in Tennessee. Black shank
affects the vascular system of the plant, with infected plants wilting
and turning yellow before dying. The stalk is blackened at ground
level and the root system is decayed; the pith in the stalk is often
separated into plate-like disks. Since the black shank fungus is often
spread through infected plants, only healthy transplants should be
used. The fungus is also spread by equipment carrying infested soil.
Equipment that has been used in contaminated fields should be
thoroughly cleaned and disinfected with a solution of one part
commercial formaldehyde in 25 parts of water (Fowlkes, et al., 1995).
Since black shank is also spread by water, plant beds and fields should
be located so they do not receive drainage from infested areas. Care
should be taken to use only noncontaminated water for transplanting
and irrigation, and to not contaminate barn areas and fields with stalks
and trash from black shank infested crops.
The level of black shank fungus in the soil is reduced, but not
eliminated, by long rotations (Lucas, 1975; Fowlkes, et al., 1995).
Some degree of loss can be expected with susceptible varieties, even
after fields have been out of tobacco for as long as 7 to 10 years
(Lucas, 1975). Black shank can best be controlled by a combination of
crop rotation, resistant varieties, sanitation and the use of soil-applied
metalaxyl (Ridomil). Resistant varieties include VA 309, VA 310, VA
331, DF 300, DF 485, TN D94, TN D950 and KY 190 (Smiley, et al.,
1986; Johnson, 1990a; Fowlkes & Miller, 1995). However, these
varieties have only a medium level of resistance to both races of black
shank. They produce increased yields in the presence of the disease
but are not immune and can suffer significant losses when levels of
black shank are high. The effective resistance of these varieties is
enhanced by the use of soil incorporated metalaxyl. The use of
metalaxyl with nonresistant varieties, however, is relatively
ineffective if significant populations of the black shank fungus are
present in the soil.
In some years, black root rot probably causes more yield reduction
than any other disease in dark tobacco. The disease, which is caused
by the soil-borne fungus Thielaviopsis basicola (Lucas, 1975), is
widepread and most destructive during cool, wet growing seasons.

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Because the symptoms associated with black root rot are not as visible
as those that occur with black shank, growers are usually unaware of
the loss that is being caused by this disease. The presence of black
root rot is indicated by slow, uneven growth of the crop. Although
some yellowing of the leaves may occur, the typical symptom is plant
stunting. Infected plants have very few feeder roots; stubs of
remaining roots are blackened and decayed. Plants may appear to
recover as the season progresses and soil temperatures begin to rise.
Black root rot can best be controlled through the use of resistant
varieties. Several varieties, including KY 171, KY 190, DF 485, DF
911, TN D94 and TN D950, have high resistance to the disease
(Smiley, et al., 1986b; Fowlkes & Miller 1995). VA 309 and VA 312
have a medium level of resistance and should be used in a 3-year
rotation with small grains or corn (Johnson, 1990a). Where
susceptible varieties are used, certain control practices will help
reduce losses due to black root rot. Dark tobacco grows well in a pH
range of 5.5 to 6.0. Since the black root rot fungus thrives at pH levels
above 6.0, growers using nonresistant varieties should attempt to
maintain pH levels between 5.5 and 6.0. Lime should be applied
according to soil test results to maintain pH levels at 5.5 or higher,
however, since other problems such as manganese toxicity can
develop at low pH levels. Cover crops should be turned under at least
3 weeks before transplanting. The breakdown of crop residues often
stimulates the growth of the black root rot fungus (Johnson, 1990a). If
tobacco is set too soon after turning under the cover crop, an increased
incidence of black root rot may occur. The application of excessive
amounts of manure and early transplanting into cold soils also favor
development of the disease.
Several other diseases cause losses in dark tobacco each year. Tobacco
mosaic virus (TMV) is one of the most easily transmitted tobacco
diseases. It causes a wide variety of symptoms on dark tobacco
ranging from a light green mottling of leaves, severe leaf mottling and
distortion, to burning of large areas of the leaves. TMV infection early
in the growing season can significantly reduce yield and quality of the
crop. Anything that moves sap from infected material to healthy
plants can transmit the disease. TMV has been known to remain alive
in cured tobacco for as long as 50 years (Fowlkes, et al., 1995). Since
the virus is present in many tobacco products, the use of these
materials should be avoided while weeding plant beds, handling
transplants or topping plants. TMV can also be spread during normal
crop cultivation. Any infected plants discovered prior to the first
cultivation should be removed from the field.
TMV can also be carried over from one season to the next in the plant
stubble or crop trash. Resistant varieties such as VA 312, DF 485, DF
911, TN D94, TN D950, KY 171 or KY 190 should be used on farms
having a history of TMV (Smiley, et al., 1986a; Johnson, 1990;
Fowlkes & Miller, 1995). The type of resistance provided by these
varieties prevents infection in the leaf. However, if resistant plants are
infected through the stalk or mid-rib, the plant will be killed. Where
mosaic resistant and susceptible varieties are being grown in the same
field, any mosaic susceptible plants should be topped first.
Damage from blue mold has not been as widespread as it has been in
other tobacco types, but some economic losses do occasionally occur
in dark fire-cured tobacco. The fungus that causes blue mold
(Peronospora tabacina) (Lucas, 1975) normally does not overwinter
in dark tobacco production areas but moves northward from other
tobacco producing regions by means of wind-borne spores. Affected
leaves first develop small yellow spots that enlarge into dead circular
areas. The undersides of these spots are covered with a bluish-gray
mold. In both the plant bed and the field, this disease is favored by
cool, rainy, overcast weather. Unprotected fields may be damaged
from the disease if spores are present when prevailing weather
conditions are favorable. Prior to 1995, blue mold was adequately
controlled by incorporating metalaxyl into the soil prior to
transplanting. However, metalaxyl tolerant strains of the fungus have
made the disease more difficult to control in recent years. Acrobat-MZ
(dimethomorph-mancozeb), a fungicide that contains dimethomorph
and carbamate, provides acceptable control of both strains of blue
mold. However, this fungicide is primarily preventive and must be
applied prior to disease incidence to be effective. Thorough coverage
of all leaves must be obtained, with sprays being re-applied every 7 to
10 days until the threat of disease development is over.
Although nematodes are not considered a problem in dark fire-cured
tobacco production in Tennessee or Kentucky, they often cause
significant yield reductions in Virginia (Johnson, 1990a). Tobacco
cyst, root knot or lesion nematodes may be present at damaging
levels, depending on geographic location and field site. A diagnostic
or predictive nematode assay should be conducted to determine if
nematodes are present in fields that are to be used for tobacco
production. Crop rotation and timely destruction of root stubble
should be practiced to keep nematode populations as low as

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possible. Selection of rotation crops is important; soybeans are
susceptible to root knot and lesion nematodes and should be avoided
in a tobacco rotation. Tomato, eggplant and pepper are also
susceptible to tobacco cyst nematodes and should be avoided
(Johnson, 1990a). Nematicides may be needed to adequately control
nematodes in some fields that have relatively heavy infestations.
Angular leaf spot, which is caused by a bacterium that results in a
brown spot on leaves during periods of rainy weather, appears to be
increasing in importance in recent years (Fowlkes, et al., 1995).
Brown spot, caused by Alternaria alternata (Lucas, 1975), can cause
significant decreases in yield and quality during some seasons.
Because the fungus damages older, mature leaves, brown spot is
primarily a problem late in the season when tobacco becomes overly
ripe. No fungicides are available for this disease, but it can be
minimized by practicing proper crop rotation and harvesting before
the crop becomes overly mature (Johnson, 1990a). Other diseases
such as tomato spotted wilt virus, target spot, hollow stalk and sore
shin cause minor damage in dark fire-cured tobacco each year
(Fowlkes, et al., 1995). Although resistant varieties or pesticides are
not available for these diseases, losses can be reduced by crop rotation
and field clean-up after harvest. Fall clean-up of tobacco fields will
help reduce the level of all diseases that affect dark tobacco. A good
job of fall clean-up involves disking plant stubble as soon as possible
following cutting, turning stubble under 2 to 3 weeks later and seeding
a small grain or fescue cover crop (Fowlkes, et al., 1995).
Insect Control
Effective insect control is essential for successful producion of dark
fire-cured tobacco. There are three general approaches to insect
control. Many producers simply follow a spray schedule and apply
insecticides every 10 to 14 days. Other growers spray only when
insects appear. The recommended method, however, is to follow an
integrated pest management approach (Semtner, 1990; Fowlkes, et al.,
1995). This method utilizes natural, cultural and chemical controls to
keep insect populations below economically damaging levels while
minimizing detrimental effects on beneficial insects and the
environment. Limited insect damage does not have an economic
impact on the crop; unnecessary application of pesticides will actually
decrease profitability. With integrated pest management, pesticides are
applied only when the economic threshold, which is the population at
which the loss from damage is equal to the cost of control, is reached
for a given insect. A successful pest management program is
dependent on accurate insect scouting practices. Tobacco fields should
be scouted for damaging insects at least once a week (Fowlkes, et al.,
1995; Pearce & Maksymowicz, 1997). Proper scouting should consist
of observations on five consecutive plants in at least 25 areas of a 1 ha
field.
Although there are several insects that occasionally damage dark
tobacco crops, only a few cause significant problems. Wireworms and
cutworms sometimes cause significant damage in newly transplanted
crops, particularly when tobacco follows sod crops. Wireworms,
which are the larval stage of click beetles, live in the soil and tunnel
into the roots and pith of tobacco transplants (Semtner, 1990).
Wireworm injury can result in stunted plant growth and reduced
stands. Because there is no remedial treatment for wireworms,
pretransplant applications of soil insecticides are recommended
(Semtner, 1990; Fowlkes, et al., 1995; Burgess & Hadden, 1996).
Treatments should be thoroughly incorporated into the soil at least 2
weeks prior to transplanting. Cutworms usually cause the most
damage to newly transplanted tobacco. They feed on the leaves and
cut off plants just above soil level; severe damage may require
retransplanting. Cutworms can be controlled by using soil-
incorporated insecticides before transplanting, or the application of
sprays or baits to the soil around the base of the plants after
transplanting (Burgess & Hadden, 1996).
Tobacco flea beetles, aphids, budworms and hornworms are the major
insect pests that feed on the leaves of dark tobacco. Extensive feeding
by flea beetles on newly transplanted tobacco causes a 'shot-hole'
effect on the leaves which may result in serious stunting. Excellent
early season control of flea beetles can be obtained by adding labeled
pesticides to the transplant water. Foliar insecticides should be applied
to newly transplanted tobacco when four beetles are observed per
plant; larger plants do not require treatment until populations reach 20
beetles per plant (Semtner, 1990).
Tobacco aphids are one of the most persistent pests in dark fire-cured
tobacco. Although aphids can be introduced into the field on infested
transplants, winged aphids that move into the field and deposit young
wingless aphids on the leaves are the most important source of
infestation (Semtner, 1990). The most severe damage occurs during
hot, dry periods of the summer. Aphids are often more severe in
partially shaded areas along the edges of fields. Aphids deposit
honeydew on tobacco leaves and a black sooty mold

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often develops. This interferes with the curing of tobacco and results
in reduced quality. Aphids cause dead areas at the base of lower
leaves which increases leaf loss at harvest. A preplant, soil-
incorporated insecticide sometimes gives aphid control for 2 to 3
months after transplanting. Research trials conducted with Admire
(imidacloprid), a new pesticide labeled in 1997, indicate that season-
long control of aphids is often obtained with this insecticide (Charles
Hadden, personal communication). If a preplant insecticide is not
used, tobacco plants should be examined regularly. The economic
threshold level for tobacco aphids is reached when 10% of the plants
have 50 or more aphids on any upper leaf. After topping, the threshold
level is 200% (Semtner, 1990; Fowlkes, et al., 1995; Burgess &
Hadden, 1996). Thorough coverage of the plant, particularly the
undersides of leaves, with a foliar insecticide is necessary to obtain
control.
The tobacco budworm female lays her eggs in the bud or on the
underside of leaves. The larvae feed on the buds of young tobacco
plants, producing holes in the developing leaves. As the leaves
increase in size, the size of the feeding holes also increases, giving the
leaves a ragged appearance. Larvae may appear in destructive
numbers any time after transplanting. Tobacco budworm control
should be initiated when five or more worms are found per 50 plants
anytime prior to the button stage of plant growth (Semtner, 1990;
Fowlkes, et al., 1995). Application of foliar sprays should be made at
4.1 bars using one or three full cone nozzles over each row. Because it
is difficult to get foliar insecticides down into the small leaves where
the worms are feeding, better control is often obtained by applying
granular baits to the plant buds (Semtner, 1990; Burgess & Hadden,
1996).
Tobacco and tomato hornworms are 2.5 to 10.0 cm long caterpillars
that consume large areas of tobacco leaves. Infestations may occur
anytime during the growing season, but the worst problems usually
occur in mid to late summer (Semtner, 1990). There are several
methods for controlling hornworms. Predators and parasites destroy
many small hornworms. On small plantings, hand picking of
hornworms may be practical. Insecticides should be applied when a
total of five or more hornworms 2.54 cm or more in length are found
on 50 plants at various locations throughout the field (Semtner, 1990;
Fowlkes, et al., 1995). Parasitized worms, which have white egg-like
cocoons on their surface, should not be included in the count. These
worms eat much less than healthy ones and provide a source of
parasites that will help reduce the next generation of hornworms
(Semtner, 1990). For best hornworm control, insecticide sprays should
be directed to the upper one-third of the plant.
Topping and Sucker Control
Dark fire-cured tobacco should be topped when the plants are in the
button to elongated button growth stage, prior to opening of blooms
(Jones, 1990a; Maksymowicz, 1994; Fowlkes, et al., 1995). Although
some growers break the tops out, it is preferable to cut the tops from
the plants with a sharp knife to decrease the possibility of disease
development in the topped plants. There is considerable variation in
topping and sucker control practices among producers. Many growers
use a sequential topping program, making two or three passes over the
field while removing the tops from only those plants that are in the
button or elongated flower stage of plant development. Other growers
choose to top the entire crop when approximately 50% of the plants
are in the early flower stage.
Research conducted at the University of Kentucky compared topping
at bud elongation, 30% bloom and full bloom (Maksymowicz, 1994).
Yield and quality were reduced as topping was delayed. The most
significant effect was the distribution of leaf within tobacco grades.
Sixty-eight percent of leaves were placed in the more desirable leaf
grade when the plants were topped at the elongated button stage,
compared to 56% when plants were topped at full bloom. Value per
hectare, based on 1991 average sale prices, was $14100 USA for bud
elongation, $12 943 USA for 30% bloom and $10326 USA for
topping at full bloom.
Plants are usually topped at about 12 to 16 leaves, with growers in
Kentucky typically topping at greater leaf numbers than Tennessee
and Virginia producers (Jones, 1990a; Maksymowicz, 1994; Fowlkes,
et al., 1995). Higher topping may result in reduced quality if barn tiers
are too close together, plants are spaced too closely in the field or
insufficient water is available during the growing season. Studies
conducted in Virginia demonstrated that extremely high topping did
not produce higher yields and proved detrimental to leaf quality
(Jones, 1990a). Research conducted in Tennessee indicated that
topping at 16 leaves resulted in higher yields than topping at the
traditional 14 leaves during seasons when soil moisture was adequate;
topping at 12 leaves resulted in significant yield reductions (Miller,
unpublished data, 1990). The quality and value of the cured leaves
were not affected by topping height. However, the upper leaves of
plants topped at 16 leaves

 Page 177
did not expand during extremely dry seasons, resulting in tobacco
with unacceptable length.
Sucker control compounds may be applied prior to topping, but they
are normally used immediately after topping is completed. Three
types of sucker control chemicals are available for use in dark tobacco
(Fowlkes, et al., 1995). Contact compounds (fatty alcohols) kill
suckers by quickly burning them and must be applied to run down the
stalk and come in contact with each sucker. Treated suckers are killed
within about one hour of application, greatly reducing the risk of
rainfall rendering the treatment ineffective. Maleic hydrazide (MH) is
a systemic sucker control compound that is absorbed into the leaves
and moves throughout the plant; it is effective when sprayed onto the
leaves and does not need to be run down the stalk. It inhibits growth
of actively growing suckers and small leaves by inhibiting plant cell
division. The only growth made after MH application is the expansion
or enlargement of cells already present in the plant. Local systemic
sucker control compounds, such as Prime+ and Butralin, also inhibit
cell division but are not translocated throughout the plant. They have
systemic activity only in the localized area of the leaf axil and must be
applied to run down the stalk like a contact compound to be effective.
Treated suckers do not turn black, but have a yellow, deformed
appearance. Local systemics will give season-long sucker control
when properly applied.
When contact sucker control compounds are used in dark tobacco,
they should be applied at the button to elongated button growth stage
(Fowlkes, et al., 1995). Suckers longer than 2.5 cm must be removed
at topping. Growers may treat individual plants as they flower or the
entire field when 50% of plants reach the elongated button stage. A
second application of a contact compound is usually made 3 to 7 days
after the first application if necessary. Contacts can be used at any
time of the day, as long as the plants are turgid and dry. They can be
applied either by hand using jugs or backpack sprayers, or with
powered spray equipment. Manual application is more time-
consuming, but generally provides better sucker control than
applications with powered sprayers. Between 644 and 859 g
chemicals are used per liter of water, with 9.45 to 19 g applied per
plant. The mixture is applied as a coarse spray so that the material
runs down the stalk. For powered spray equipment, a 4% solution is
used (18.7 L of chemical with 468 L of water). Approximately 468 L
of mixture, applied at 1.4 bars, are applied per hectare. Three solid
cone nozzles per row are recommended to ensure proper application.
Two TG-3 nozzles should be used on the outsides of the row, with a
TG-5 nozzle used for the center (Fowlkes, et al., 1995). The outside
nozzles should be spaced approximately 23 cm from the center one
and angled at 45°; all three nozzles should be directed at the center of
the row, 30.5 to 40.6 cm above the top of the plants. Additional
applications of contact, systemic or local systemic compounds are
usually required to prevent late season sucker growth.
Many dark tobacco producers prefer to use local systemics because
they give season-long sucker control. Because these chemicals are not
translocated throughout the plant, they must be applied in a manner
similar to contact compounds (Fawlkes, et al., 1995). Plants should be
topped, with suckers longer than 2.54 cm removed, at the elongated
button stage. The entire field may be topped at once, or plants may be
topped individually during two to three passes across the field.
However, individual plants should not be treated with a local systemic
more than once. Because application of local systemics to plants that
have not reached the elongated button stage may cause distortion of
immature leaves, many growers use these chemicals following an
earlier application of a contact sucker control compound or as a tank-
mix with MH. When applied alone, local systemics are mixed as a 2%
solution (Fowlkes, et al., 1995). Application can be made by hand or
by powered spray equipment as described for contact compounds.
However, because carry-over residues may damage small grain cover
crops, smaller volumes of these solutions are applied per plant. When
hand applied, 9.5 to 14.2 g of solution is used per plant; with power
sprayers, 281 L per hectare are applied. Thorough coverage of every
sucker is essential, since untreated suckers will grow vigorously. Rain
occurring within 2 hours after spraying will significantly reduce
effectiveness.
Although MH is labeled for use in dark tobacco, it is not commonly
used as a stand-alone treatment (Fowlkes, et al., 1995). When applied
to actively growing dark tobacco plants, MH may discolor the top
leaves, reducing their quality; leaves less than 20 cm in length will
likely be stunted. If MH is used in dark tobacco, it is typically used
following the application of a contact or local systemic, or as a tank-
mix with a local systemic. When used alone or as a sequential
treatment, MH is applied at the rate of 14 to 19 L/ha, applied in 151 to
189 L of water. Because MH is absorbed and translocated throughout
tobacco plants, it does not need to touch each sucker bud to be
effective. Application as a fine mist to the upper one-third to one-half
of the plant at a pressure of 2.8 to 3.5 bars is sufficient.

 Page 178
Effectiveness of MH is improved by treating plants while leaf stomata
are open; if possible, sprays should be applied on cloudy days or early
in the morning after the dew has dried. Application of MH during the
middle of hot days reduces chemical absorption and increases the risk
of leaf burn. Plants under drought stress do not absorb and translocate
MH readily, which reduces the effectiveness of sucker control in dry
growing seasons. Rainfall occurring within 6 hours of MH application
may reduce effectiveness and require respraying. If rain occurs within
3 hours after spraying, a full rate of MH should be used during
respraying (Fowlkes, et al., 1995). Rains occurring within 3 to 6 hours
after application require only a half rate during respraying.
One of the most effective sucker control programs combines the use
of contact compounds at topping, followed 5 to 7 days later with a
tank-mix treatment of a local systemic and MH (Fowlkes, et al.,
1995). This combination prevents stunting of leaves that are immature
at topping and provides season-long sucker control. The tank mix
contains a half rate of either Prime+ or Butralin combined with a full
rate of MH. The tank mix is applied with powered spray equipment as
described for the mechanical application of contact sucker control
compounds.
Harvesting and Curing
The production of a good crop of dark firecured tobacco in the field
does not automatically result in a quality product at marketing. Many
excellent crops are ruined during the harvesting and curing process.
Methods used for harvesting and curing vary considerably among
production areas and among individual growers within the same
neighborhood. The techniques that are utilized for one crop, or during
a particular curing season, may be inappropriate for another crop or
season. However, there are several basic principles that are involved
in harvesting and curing dark firecured tobacco.
Harvesting dark tobacco at the proper maturity stage is essential for
producing a quality product (Everette, 1968; Rhodes, et al., 1980;
Jones, 1990b). Mature tobacco cures much more easily, and usually
with a better color, than tobacco that is green or over-ripe. Tobacco
that is housed green will not cure properly, while over-ripe tobacco
loses desirable body and quality (Everette, 1958). Dark tobacco
normally ripens 3 to 5 weeks after topping. Mature leaves will droop,
thicken and become oily. Dark tobacco is ready for harvest when the
leaves begin to lose their dark green color and take on a slight
yellowish cast. Many growers determine leaf ripeness by folding a
tobacco leaf. Ripe leaves will crack readily when creased between the
fingers.
Dark firecured tobacco is stalk harvested. Research conducted at the
University of Kentucky during the late 1960s comparing stalk
harvesting with leaf priming (Everette, 1968) indicated that primed
leaves could be cured as well as those remaining on the stalk. Cured
leaves that had been primed were judged to have equal or better
quality and higher elasticity than traditionally cured leaves, but they
were thinner bodied and less desirable for some segments of the
tobacco trade. Traditionally, dark firecured tobacco producers in
Virginia have primed the bottom two to three leaves before stalk
harvesting. However, studies investigating the effects of priming
conducted at the Southern Piedmont Center in 1979 (Jones, 1990b)
and at the University of Kentucky in 1979 to 1980 (Miller & Everette,
unpublished data, 1979) demonstrated that priming of dark tobacco is
not economically feasible.
The most common method of harvesting dark firecured tobacco is to
cut the stalks 5 to 10 cm above the ground (Everette, 1958; Jones,
1990b; Fowlkes, et al., 1995). Some growers will split the stalk down
to about 15 to 20 cm from the ground before cutting. Although
splitting the stalk increases the rate of moisture loss and promotes
rapid curing, there is increased risk of excessive leaf breakage unless
workers are skilled in this practice (Everette, 1958). Regardless of
whether the stalk is split, it is important to wilt the tobacco to
minimize leaf breakage and to make it easier to handle and cure.
However, great care must be taken to prevent the tobacco from
scalding during wilting. In some production areas, cut plants are
turned upside down and allowed to wilt before being placed on the
stick. The amount of time required for proper wilting is dependent on
the weather. One to three hours are usually sufficient under warm,
clear weather conditions. Once the tobacco has wilted, five to seven
plants are placed on a stick, depending on the size of the plant.
During hot weather, it is difficult to wilt tobacco in this manner
without the occurrence of significant scalding. As a result, many
growers place the plants directly onto sticks as they are cut. For either
cutting method, the tobacco needs to remain in the field for an
additional 24 to 48 hours to allow further wilting (Everette, 1958;
Fowlkes, et al., 1995). This is best accomplished by scaffolding the
tobacco. Scaffolding greatly decreases the possibility of scalding,
particularly if the tobacco is covered with burlap, and minimizes

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damage from rains that may occur. Fixed wooden or rolling scaffolds
work equally well. Rolling scaffolds are more efficient, but are
sometimes cost prohibitive due to the large number that are required.
When wooden scaffolds are used, the tobacco is transferred to wagons
or rolling scaffolds for transport to the barn.
After the tobacco has properly wilted in the field, it is ready for
transport to the curing barns. The location and design of the barn has a
significant influence on the proper curing of dark fire-cured tobacco
(Everette, 1958). Barns are often located in a wooded or sheltered area
to minimize extreme fluctuations in temperature and humidity during
curing. Barns are constructed to be virtually airtight, with ventilators
placed near the bottom and at the top of the structure to control
humidity. A tight barn holds heat and moisture when desired and
greatly minimizes the risk of fire destroying the structure.
A barn fire, resulting in the loss of both the crop and the building, is a
dark tobacco producer's worst nightmare. Many barns have a concrete
foundation that extends 0.9 to 1.2 m up the side walls to decrease the
chance of the barn catching fire. The doors on the bottom ventilators,
which normally run the entire length of the barn, are positioned to
direct air above the fires; this decreases the chance of sparks igniting
the dried leaves. In spite of these precautions, it is not unusual for a
few barns to be destroyed by fire during a typical curing season.
It is important to completely fill a barn as quickly as possible to
promote uniform curing (Everette, 1958). Sticks are normally spaced
20 to 35 cm apart, depending on the size of the tobacco and the time
of season; early harvested tobacco requires more space than tobacco
harvested later in the season. It is important that the tails of tobacco on
one tier do not overlap the butts of plants on the tier below. Very large
tobacco may require that only alternating tiers be used. The stalks on
each stick are evenly spaced to promote uniform air and smoke
movement during curing. This minimizes problems with houseburn
and facilitates an even, desirable cure of the tobacco.
The most difficult aspect of dark tobacco production is the curing of
the leaves. This process involves the proper, timely manipulation of
humidity, heat and smoke (Everette, 1958; Rhodes, et al., 1980).
Many chemical changes take place during the curing process. These
changes are highly dependent on temperature. If temperatures drop
below 15.6°C, the tobacco will continue to dry but will not cure
properly (Everette, 1958). Temperatures above 37.8°C may result in
the development of undesirable leaf colors.
Although considerable research has gone into 'scientifically' curing
dark tobacco, the process remains more art than science, with specific
techniques varying significantly among growers. Curing of dark
tobacco is highly dependent on the weather, tobacco variety and the
size and quality of a given crop. No two curing seasons are exactly
alike; this makes the experience of the individual dark tobacco
producer an essential component of the curing process. However,
there are several critical stages that each crop must go through to
achieve a quality product. These stages can generally be described as:
(1) yellowing the leaves,
(2) setting the leaf color,
(3) drying down the stalks and leaves, and,
(4) applying 'finish' to the leaves (Everette, 1958; Rhodes, et al., 1980;
Jones, 1990b; Fowlkes, et al., 1995).
The degree of yellowing obtained before the fires are started has a
pronounced influence on leaf color. Ideally, leaf yellowing should be
obtained without the use of fires and with complete ventilation
(Everette, 1958). Proper yellowing will usually occur within 5 to 8
days. However, the use of high nitrogen rates, limited barn space and
adverse weather conditions often necessitate the use of fires during the
yellowing phase of curing. Although fires are not normally used until
yellow spots begin to appear in the leaves, small fires may be
necessary to achieve proper yellowing if temperatures drop below
21°C. During warm, wet weather, open dry fires may be needed to
prevent houseburn.
When yellowing is completed, the tobacco is ready for firing. All
ventilators, except those in the top of the barn, are closed and the fires
are ignited. Two general methods of building fires are currently used,
depending primarily on the production area (Everette, 1958; Rhodes,
et al., 1980). The first method uses hardwood logs that are 0.9 to 1.2
m in length and 10 to 25 cm in diameter. Several piles, 1.8 to 2.4 m on
center, are positioned over the entire barn floor and covered with
sawdust. A small hole is left uncovered for starting the fire. The other
method of building fires is to lay hardwood slabs end-to-end for the
entire length of the barn. The rows of slabs, which are spaced 1.8 to
2.4 m apart, are covered with sawdust, with openings left at various
places for starting the fires. Kerosene and kindling are used to start the
fires. The amount of sawdust that is used during the first firing
depends on the amount of moisture present in the tobacco, the relative
humidity and the temperature. Relatively small

 Page 180
fires are normally used to increase the temperature to approximately
27° to 32°C. Hotter fires may be needed if excessive moisture is
present in the crop. However, temperatures should not exceed 38°C to
prevent the development of undesirable colors in the cured leaf. The
relative humidity should be maintained at approximately 85 to 90%.
After the first firing is started, it should continue until desirable color
has developed in the leaves. This process will normally take
approximately 6 to 8 hours, but may vary considerably under differing
firing conditions. When the color is properly set, the mid-rib of the
leaf will still be green, but the lamina will be uniformly brown.
Once the color has been set, drying of the leaves and stalks begins
(Everette, 1958; Jones, 1990b; Rhodes, et al., 1980). Good ventilation
is provided and the amount of heat is increased. The relative humidity
should be decreased to about 75 to 80%. The drying process is
continued until the leaf is dry and the lamina cracks when touched.
This will usually take from 7 to 14 days, depending on prevailing
curing conditions. When the crop has been adequately dried, the
bottom half of the mid-rib will normally have turned brown. The barn
is then ventilated and the leaves are allowed to come into 'case' or
'order' (Everette, 1958; Rhodes, et al., 1980; Jones, 1990b). This
means that the leaves have absorbed sufficient moisture from the air to
become soft and pliable. This is critical because it allows the color
that has developed in the leaf to 'run' or become more uniform. This
sequential process of firing the tobacco, followed by allowing it to
come into order, may need to be repeated several times to achieve the
desired color in the leaves. The stems and veins are also darkened
during this process.
The development of satisfactory color is much easier during some
curing seasons than others. During adverse curing seasons, the skill of
the grower is extremely critical in the production of a quality crop. In
dry seasons, it may be necessary to wet the floors of the barn, or the
sawdust itself, to facilitate proper curing. Under wet curing seasons,
houseburn may be a serious problem. This is often referred to as
'strutting' or 'sweating' (Everette, 1958; Rhodes, et al., 1980; Jones,
1990b). Sweating is caused by water condensing on the curing leaf.
The water-soaked areas will cure black rather than brown. When the
cured leaf is stretched, sweated areas will often tear. In severe cases of
sweating, the leaves may rot and drop from the stalk. Sweating often
occurs during the first few days of firing when moisture is high; it can
be corrected by adding heat and increasing ventilation in the barn.
The final stage of the curing process is adding 'finish' to the crop.
Finish refers to smoke deposits on the leaf which give it good aroma
and a shiny, somewhat gummy texture (Everette, 1958; Jones, 1990b).
This is achieved by repeated firing using low fires with a lot of
sawdust and minimum ventilation. The objective is to maximize the
smoke level in the barn and to minimize the temperature. Finishing
the crop is much easier in damp weather. Many growers periodically
let the fires go out so that the tobacco will come in case. Other
producers keep the fires going continuously, but wet the floors and the
sawdust to increase the humidity in the barn. The length of time and
number of firings required for finishing the crop will vary among
growers and from season to season.
Market Preparation
When the crop has been cured and finished, the tobacco is taken down
from the rails and bulked. This is normally done while the tobacco is
in good case and soon after the last firing to prevent loss of aroma.
The tobacco is placed on a platform prepared by laying down rows of
boards supported by logs or blocks, with at least 7 to 10 cm of air
space underneath the bulk. Bulking is done by taking the stalks off the
sticks and placing the butts out on alternating sides of the pile
(Everette, 1958). The finished bulk is covered with plastic or a
tarpaulin, with the butts uncovered. Ideally, tobacco is left in the bulk
at least 2 to 4 weeks before stripping, since color and aroma improve
during bulking.
Unlike most other tobacco types, a substantial portion of the dark fire-
cured tobacco crop is sold in the barn each year and delivered directly
to the buyer. In these situations, the way in which the tobacco is
handled during stripping will vary with the particular grower and
buyer. However, regardless as to whether the crop is sold directly at
the farm or at a warehouse, it is important that growers properly
prepare dark tobacco for market.
Many growers grade dark tobacco into only three classes. However, in
recent years nondescript and green grades of dark tobacco have often
brought only the support price. For this reason, the more traditional
practice of making 'trash lugs' and 'out leaf grades has been more
profitable. Typical farm grades for a well cured crop of dark tobacco,
topped at about 14 to 16 leaves, should include lugs, seconds, leaf and
out leaf (Fowlkes, et al., 1995). Lugs usually include the bottom three
to five leaves. These leaves are thin-bodied and may be injured, torn
or dirty. This grade can be

 Page 181
improved by splitting it into good lugs and trash lugs, based on the
amount of dirt and injury present. The seconds grade usually consists
of the next three to five leaves on the stalk. This grade is separated
primarily according to the body of the leaf. Color and finish may be
about the same as that of the leaf grade. The leaf grade is usually
made up of the top four to six leaves on the stalk. These leaves are the
most desirable and will have good body, elasticity and plenty of
finish. In some production areas, growers also make a 'wrapper' grade,
which is very high quality leaf used in the manufacture of specialty
cigars. The final grade should be the out leaf. This throw-out grade is
used for leaves from the seconds and leaf positions which have green
areas in them or are otherwise undesirable.
As the tobacco is stripped, it is tied in small, neat hands comprising
about 8 to 12 leaves (Everette, 1958; Rhodes, et al., 1980; Fowlkes, et
al., 1995). After several hands have been tied, they are bulked three to
four hands at a time. Several methods of bulking are used, depending
upon how the tobacco will be marketed. If the tobacco is marketed at
a warehouse, individual grades may be bulked directly onto marketing
baskets at the barn, or bulked in wooden frames and then re-bulked
onto baskets at the warehouse. Each basket of tobacco will be given
an official standard grade by USDA graders based on quality, group,
color and length of leaf and assigned the appropriate support price.
The tobacco is sold through an auction system similar to that used for
burley and flue-cured tobacco. The dark firecured tobacco producers'
long, laborious task is then completed just in time to begin the next
season's crop.
The reader is referred to Chapter 14 for additional comments on dark
air-cured tobaccos.
References
Agricultural Marketing Service (1996) Tobacco Market Review:
FireCured and Dark Air-Cured, 1995 Crop. USDA. US Government
Printing Office, Washington, DC.
Burgess, E. & Hadden, C. (1996) Tobacco pest control. Agric. Ext.
Serv. Pub. SP 91. University of Tennessee, Knoxville.
Chaplin, J.F., Baumhover, A.H., Bortner, C.E., et al. (1976) Firecured
tobacco. In: Tobacco Production. USDA-ARS Agric. Info. Bull. No
245. Washington, DC.
Duncan, G. & Anderson, B. (1997) Protecting greenhouse plants. In:
Burley and Dark Tobacco Production Guide 1997. Rural Press USA,
Raleigh, NC.
Everette, G. (1958) Harvesting, curing and preparing dark fired
tobacco for market. Agric. Ext. Serv. Circ. 555. University of
Kentucky, Lexington.
Everette, G. (1968) Evaluation of primed versus stalk cured leaf of
dark firecured tobacco. Tob. Abstr., 12, No 11, abstr. 2593, 825.
Fowlkes, D.J. (1989) Producing tobacco transplants. Agric. Ext. Serv.
Pub. PB 1385. University of Tennessee, Knoxville.
Fowlkes, D.J. (1997) The float system for producing tobacco
transplants. Agric. Ext. Serv. Pub. 167. University of Tennessee,
Knoxville.
Fowlkes, D.J. & Miller, R.D. (1995) Dark tobacco variety
information 1995 Tennessee recommendations. Agric. Ext. Serv. P &
SS Info. No 215. University of Tennessee, Knoxville.
Fowlkes, D.J., Miller, R.D., Jared, J.R., et al. (1995) Dark tobacco
production in Tennessee. Agric. Ext. Serv. Pub. 899. University of
Tennessee, Knoxville.
Hourigan, W.W. (1986) Federal programs: burley and dark leaf
controls. In: Tobacco in Kentucky. Kent. Ext. Serv. Pub. ID-73.
University of Kentucky, Lexington.
Hunter, P.P., Jones, P. & Hilty, J.W. (1981) The occurrence and
distribution of races of Phytophthora parasitica var. Nicotianae in
dark tobacco in Tennessee. Tob. Sci., 25, 2021.
Johnson, C.S. (1990a) Disease control for dark-fired tobacco. In: 1990
Dark FireCured Tobacco Production Guide. Va Coop. Ext. Serv. Pub.
436-048. Virginia Poly. Inst. and State Univ., Blacksburg.
Johnson, C.S. (1990b) Weed control for dark-fired tobacco. In: 1990
Dark FireCured Tobacco Production Guide. Va Coop. Ext. Serv. Pub.
436-048. Virginia Poly. Inst. and State Univ., Blacksburg.
Jones, J.L. (1990a) Agronomic information. In: 1990 Dark FireCured
Tobacco Production Guide. Va Coop. Ext. Serv. Pub. 436-048.
Virginia Poly. Inst. and State Univ., Blacksburg.
Jones, J.L. (1990b) Harvesting and curing. In: 1990 Dark FireCured
Tobacco Production Guide. Va Coop. Ext. Serv. Pub. 436-048.
Virginia Poly. Inst. and State Univ., Blacksburg.
Latham, D.H. (1969) DF 300: a black shank resistant dark firecured
tobacco and observations on tobacco black shank in Tennessee. Agric.
Exp. Sta. Bull. 453. University of Tennessee, Knoxville.
Legg, P.D. & Miller, R.D. (1988) Cultivar differences and genotype ×
year interactions in dark tobacco under two topping systems. Tob.
Sci., 32, 2932.
Legg, P.D. & Tedford, E. (1989) Registration of 'KY 190' Tobacco.
Crop Sci., 29, 1093.
Lucas, G.B. (1975) Diseases of Tobacco. Harold E. Parker & Sons,
Fuquay-Varina, NC.
Maksymowicz, B. (1994) Dark tobacco production guidelines. In:
Burley and Dark Tobacco Production Guide 1994. Rural Press USA,
Raleigh, NC.
Miller, R.D., Eggett, B.C. & Hensley, R.A. (1995) Registration of 'TN
D94' dark firecured tobacco. Crop Sci., 35, 1712.
Miller, R.D., Eggett, B.C. & Hensley, R.A. (1998a) Registration of
'TN D950' dark firecured tobacco. Crop Sci. (in press).
Miller, R.D., Fowlkes, D.F. & Buschermohle, M.J. (1998b) Producing
tobacco float plants by direct seeding in outdoor water beds. Agric.
Expt. Sta. Res. Rept. University of Tennessee, Knoxville (in press).
Miller, R.D., Hensley, R.A. & Wilhoit, U.D. (1994) Instructions

 Page 182
for building poor-boy tobacco seeders. Agric. Expt. Sta. Res. Rep.
94-04. University of Tennessee, Knoxville.
Miller, R.D. & Hunter, P.P. (1987) Registration of 'DF 485' dark fire-
cured tobacco. Crop. Sci., 27, 131314.
Miller, R.D. & Legg, P.D. (1990) A grade index for type 22 and type
23 fire-cured tobacco. Tob. Sci., 34, 102104.
Parks, W.L. & Safley, L. (1967) Effect of soil pH and potash on the
yield and quality of dark tobacco. Agric. Expt. Sta. Bull. 414.
University of Tennessee, Knoxville.
Pearce, B. (1997) A closer look at plant nutrients. In: Burley and Dark
Tobacco Production Guide 1997. Rural Press USA, Raleigh, NC.
Pearce, B. & Maksymowicz, B. (1997) Careful management needed
with float plants. In: Burley and Dark Tobacco Production Guide
1997. Rural Press USA, Raleigh, NC.
Rhodes, G.N. & Fowlkes, D.J. (1995) Control weeds in burley and
dark tobacco. Agric. Ext. Serv. Pub. PB 1071. University of
Tennessee, Knoxville.
Rhodes, G.N., Hunter, P.P. & Burgess, E.E. (1980) Dark tobacco
production in Tennessee. Agric. Ext. Serv. Pub. 899. University of
Tennessee, Knoxville.
Semtner, P.J. (1990) Insects on tobacco. In: 1990 Dark Fire-Cured
Tobacco Production Guide. Va Coop. Ext. Serv. Pub. 436-048.
Virginia Poly. Inst. and State Univ., Blacksburg.
Shuffett, D.M. (1986) Tobacco's economic impact in Kentucky. In:
Tobacco in Kentucky. Kent. Ext. Serv. Pub. ID-73. University of
Kentucky, Lexington.
Smiley, J.H., Nielsen, M.T. & Palmer, G. (1986b) Variety selections
very important. In: Tobacco in Kentucky. Kent. Ext. Serv. Pub. ID-73.
University of Kentucky, Lexington.
Smiley, J.H., Palmer, G. & Calvert, J.R. (1986a) The seed bed. In:
Tobacco in Kentucky. Kent. Ext. Serv. Pub. ID-73. University of
Kentucky, Lexington.

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Chapter 6
Major Tobacco Diseases
6A
Fungal and Bacterial Diseases
P.B. Shoemaker and H.D. Shew
North Carolina State University
Raleigh, North Carolina, USA
Introduction
There is a large number of diseases caused by fungi and bacteria that
affect tobacco. Less than 25% of these are considered major diseases
which will be discussed here. Some fungal and bacterial diseases
occur on the leaf and thus have a direct effect in reducing yield and
quality (eg. blue mold, brown spot, powdery mildew, target spot,
wildfire and angular leaf spot). Some fungal diseases cause root rots,
which result in stunting, and thus indirectly reduce yield and quality
(e.g. black shank and black root rot). Other fungal and bacterial
diseases may infect the plant stalk or vascular system and result in
death and loss of the entire plant before it is harvested (e.g. black
shank, sore shin, Granville wilt, collar rot, Fusarium wilt and hollow
stalk). For more detailed information and coverage of the many
tobacco diseases, the reader may consult such references as Diseases
of Tobacco: Compendium of Tobacco by G.B. Lucas.
Tobacco Diseases
a
Blue Mold
Blue mold can be one of the most destructive diseases affecting
tobacco. It is most damaging following extended periods of wet and
rainy weather during which the fungus has had an opportunity to
produce several cycles of spores. Depending on weather conditions,
growth stage and other factors, losses from blue mold can range
between only slight to complete destruction of the crop in a field. The
pathogen which causes blue mold is present in many tobacco
production regions, including the Americas, the Euro-Mediterranean
region and Australia. However, the disease has not been reported
south of the equator in Africa nor in east Asian countries such as
China and Japan.
Symptoms
Symptoms vary depending on the age of the plant at the time of
infection. Seedlings that are infected within 2 to 3 weeks of
germination will yellow, shrivel and die. Older seedlings may show
symptoms of either systemic infection or leaf infection. When
seedlings are systemically infected, the leaves have a twisted and
puckered appearance and the terminal bud stops growing. Suckers
may begin to grow in leaf axils, and in severe cases the seedling dies.
The most characteristic symptoms of blue mold are the leaf spots,
which may develop on leaves of seedlings or leaves of any age
following transplanting. Blue mold leaf spots are circular and appear
yellow on the upper leaf surface and bluish and moldy on the lower
leaf surface, hence the name of the disease. Usually, the yellow leaf
spot will appear the day before the bluish mold appears. The yellow
leaf spot with the bluish mold will maintain this appearance for about
a week, after which the leaf tissue in the spot becomes necrotic and
tan in color and the mold turns brown. Later, the tissue in the leaf spot
may fall out of the leaf leaving an empty hole 2 to 4 cm in diameter. If
infection occurs during the first 6 to 8 weeks following transplanting
and leaf spots are very numerous, the infected leaves will become
ragged in appearance and eventually blight and fall off the plant. If
infection occurs later in the growing season, the leaves are likely to be
retained on the plant and the leaf spots will be very evident following
harvest and curing. Another symptom of blue mold results when
infections

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occur on leaf veins which are referred to as locally systemic
infections. These result in a slight yellowing and distortion at the point
of infection and are rarely accompanied by sporulation of the fungus.
Plants that survive early systemic infection may appear to develop
normally but are usually stunted and subject to late season lodging.
Cutting across the stem of systemically infected plants will reveal a
reddish-brown discoloration in the vascular tissue.
Pathogen Description
The blue mold fungus is currently referred to as Peronospora
tabacina Adam, but the name Peronospora hyocyami de Bary may
have precedence. The fungus is in the class of fungi Oomycetes
commonly referred to as water molds. The blue mold fungus is an
obligate parasite which means it will grow and reproduce only in
living tissues. It produces two types of spores: asexual spores called
sporangiospores and sexual spores called oospores. Sporangiospores
are the spores produced in the bluish mold on the underside of the leaf
spot. These spores are produced in abundance with as many as one
million spores per square centimeter of infected tissue possible. Under
the microscope, sporangiospores appear as small, thin-walled lemons
which are clear in color and approximately 15 × 25 mm in size.
Sporangiospores are the principal means by which the disease is
spread. However, these spores are relatively shortlived. Direct
exposure to sunlight will kill most within an hour, but under continual
high humidity and cloudy conditions sporangiospores will survive for
several days. Oospores, in contrast, are produced within the infected
tissue. They have a spherical shape, are thick-walled and reddish-
brown and range in size from 20 to 60 mm diameter. To date, there is
very little evidence that oospores germinate or are involved in survival
or carryover of the fungus from season to season.
Epidemiology
Blue mold epidemics develop when P. tabacina and the tobacco plant
are present together and weather conditions favor disease
development. In tropical and subtropical regions, the blue mold
pathogen and cultivated tobacco and/or wild tobacco species are
present together throughout the year and blue mold epidemics are
common. In contrast, in temperate regions the tobacco crop is
seasonal, the blue mold pathogen is not believed to overwinter and
blue mold epidemics are less common. In both regions, however,
when blue mold epidemics do appear, they can be equally destructive.
Chronological maps of past epidemics show that blue mold in some
years advances with successive crops as they are planted.
Sporangiospores are readily released from blue mold infected leaves
and are capable of being windblown for several kilometers to infect
tobacco plants in distant fields. In other cases, blue mold has shown
up in areas hundreds of kilometers from any reported outbreaks of the
disease, indicating long distance transport of sporangiospores. Besides
movement of spores on air currents, the disease may also be moved on
infected transplants. Epidemics initiated from infected transplants may
begin earlier, develop faster and be more destructive than those
initiated by windblown spores.
Once P. tabacina has been introduced into a tobacco field, further blue
mold development is very dependent on subsequent environmental
conditions. If conditions are sunny and dry, the disease will fail to
develop in open fields. If conditions, on the other hand, are relatively
cool, overcast and wet, a raging epidemic may develop. In the
presence of free water, sporangiospores will germinate and infect the
leaf in 2 to 4 hours Symptoms and a new crop of sporangiospores will
appear 5 to 7 days later. Maximum disease and sporulation occurs if
temperatures during this time have been between 15 and 25°C. A
precondition for sporulation is that the relative humidity must be
greater than 95% for at least three hours the night before
sporangiospores develop. The spores are released in the hours before
noon as humidity drops and air temperatures rise. Spores that are
released and exposed to direct sunlight are readily killed. If spores
land on tobacco leaves where they are shaded, they may survive until
nightfall when free water from rain or dew is a prerequisite for
germination and infection. Spores that are released during overcast
and wet weather have a far greater chance of surviving and
successfully initiating new infections. During extended periods of
cool, cloudy and wet weather, P. tabacina can produce several cycles
of new spores. Nearly total crop loss can occur in as short as a 3-week
period during blue mold favorable weather.
Management
The best strategy to manage blue mold is to prevent the early
introduction and establishment of P. tabacina. It is essential that
seedlings be protected by the preventive use of fungicides during all
phases of transplant production. This may be followed, in many
countries, by preventive use of certain fungicides in the field. The
choice of which fungicides to use is dependent on

 Page 185
governmental regulations, whether or not fungicideresistant P.
tabacina strains are present and the need to prevent undesired
fungicide residues on the harvested leaf.
Seedlings should be sprayed every 5 to 7 days with a protectant
fungicide such as mancozeb, and care should be taken to assure
complete coverage. Recent labeling of metalaxyl in the United States
prohibits its use during any tobacco transplant production for fear of
early selection of metalaxyl resistant strains of the fungus. Metalaxyl
is labeled as a soil preplant incorporated treatment for field use in the
United States and as a prepack mixture for spraying on the leaves in
many other countries. Metalaxyl has been an extremely effective
fungicide for sensitive strains of P. tabacina, but resistant strains are
becoming more prevalent. Registrations are being sought for
alternative fungicides, such as dimethomorph, which have no cross
resistance with metalaxyl. As new fungicides are developed and made
available, there should be care taken in their use to prevent
development of multiple fungicide resistance.
Currently, there are no commercial cultivars with resistance to blue
mold. Efforts are underway at North Carolina State University to
develop resistance in burley and flue-cured types, utilizing a source of
resistance from Ovens 62. Two breeding lines, NC-BMR-42 and NC-
BMR-90, were released in 1997 and efforts continue to develop
resistant tobacco cultivars through back-crossing.
Blue mold is a disease that lends itself to regionalwide management.
When the disease occurs in Turkey or Greece, it is of concern in
France and Germany. Likewise, when blue mold occurs in the
Caribbean or Mexico, it is of concern in the United States. Blue mold
warning services are operated by the Centre de Coopération pour les
Recherches Scientifiques au Tabac (CORESTA) headquartered in
Paris for the European and Mediterranean countries and by the North
America Blue Mold Forecast System located at North Carolina State
University for North America. Blue mold coordinators in each country
or state send blue mold status reports to the central office, which in
turn issues blue mold warnings to all the participants. The computer-
based Blue Mold Forecast System for North America was placed on-
line on the Internet in March 1996. This system can be accessed from
anywhere by way of the worldwide web. It provides. expected
trajectories of spore movement from known sources of blue mold
based on 48-hour weather forecast models. Blue mold management in
the future could become much more sophisticated with the full
integration of computer assisted forecast and warning systems.
b
Brown Spot
Brown spot can be a serious disease on all tobacco types and has been
reported from most tobacco producing countries. It can cause direct
loss to leaves by the production of leaf spots, by causing leaves to
drop off the plant before they are harvested or by causing harvested
leaves to shatter during handling. Brown spot infection also affects
leaf quality by alterations in chemical constituents of the leaf. For
example, sugar/nitrogen and sugar/alkaloid ratios decrease and
nitrogen/alkaloid ratios increase with increased disease severity
(Lucas, 1975).
Symptoms
There are several leaf spots that can affect tobacco, and the untrained
observer might easily confuse them. The brown spot disease causes
very conspicuous brown circular spots, approximately 1 to 3 cm in
diameter, which are surrounded by a definite yellow halo. Black
concentric rings are present within the leaf spots. Leaf spots may form
on any leaves, but they are most prevalent on the uppermost leaves
late in the growing season. Leaves with heavy infection may abscess
and fall to the ground.
Pathogen Description
Brown spot is caused by the fungus Alternaria alternata (Fr. ex Fr.)
Keissl. The fungus is in the class Hyphomycetes, a group of fungi for
which no sexual state has been found. Alternaria alternata produces
asexual spores called conidia, which look like small clubs under the
microscope. The conidia are multicelled, with a swollen base that has
cross walls and usually a tip cell called a beak. They are brown to
olive in color and 18 to 47 × 7 to 18 mm in size (Lucas, 1975). The
conidia are produced on short stalks called conidiophores, which arise
from the concentric rings that can be observed on the surface of the
leaf spot.
Epidemiology
Fields that are used continuously for tobacco production are at greater
risk of having problems from brown spot than fields where crop
rotation is practiced. Ahernaria ahernata can survive, grow and
reproduce in tobacco debris left from a previous crop. Following
transplanting, conidia produced on carried over debris

 Page 186
can be splashed up onto lower leaves. Moisture, in the form of rain or
dew, is a prerequisite for germination. In the presence of moisture,
conidia will begin to germinate within an hour and the resulting germ
tubes will penetrate the leaf either directly or through stomares. A new
crop of conidia can begin to develop in 7 to 8 days, but longer
incubation periods are common. Several disease cycles occur within a
growing season, and spores can be splashed or wind-blown from plant
to plant. Disease build-up is progressive, with little damage noticeable
for the first 2 months following transplanting and suddenly becoming
very obvious the final 3 to 4 weeks. Heavy rains and dews later in the
growing season can result in severe damage. Plants also are
predisposed to heavier damage from brown spot by plant stress that
may be caused by drought, drowning, nematode infection or
imbalanced fertilization. Brown spot is more severe under conditions
of either phosphorus or potassium deficiency or excess nitrogen
fertilization (Lucas, 1975).
Management
Brown spot is best controlled by following sound cultural and
production practices. Fields in which brown spot was severe one year
should not be planted to tobacco the following year. Also, fields
known to be infested with root knot nematode should be treated or not
used for tobacco. Fertilization should be based on soil test
recommendations, and excess nitrogen should be avoided. Plants
should be topped and treated for suckers in a timely manner. Delaying
topping and allowing sucker growth increase the susceptibility of the
plant to the brown spot disease.
Some burley and flue-cured cultivars are reported to have some
tolerance to brown spot (Lucas, 1975). There are no commercial
cultivars which have been bred specifically for resistance to brown
spot. Fungicides, such as maneb, mancozeb, anilazine and iprodione,
are very effective against brown spot. However, because of concerns
of fungicide residues that would be left on the leaf, the use of
fungicides have not been approved for this purpose. Currently, there is
considerable activity in the development of new fungicide chemistry.
If a highly active compound for Alternaria were found that was safe
and left negligible residues, then fungicidal control of brown spot
could become a possibility.
c
Powdery Mildew
Powdery mildew can cause major losses to tobacco when it occurs. It
is a sporadic disease, that is most important in tobacco producing
countries in southern Africa, the Mediterranean and Eastern Europe. It
occurs in eastern Asia and the Americas but not in the United States.
When the disease is severe, powdery mildew can completely cover the
leaf surface and render the leaf unusable.
Symptoms
The major symptom of powdery mildew is the mold produced by the
causal fungus. The powdery mildew fungus grows and reproduces on
the leaf surface where the visible vegetative mycelium and
reproductive spores form a cotton-like web, hence the name powdery
mildew. The grayish-white mildew appears about 6 weeks after
transplanting as small tufts on fully expanded leaves. The mildew
does not usually develop on young leaves. It develops first and more
rapidly on the under leaf surface, but eventually covers the upper leaf
surface as well. Also, brown spots may develop in the tobacco tissue
on the upper leaf surface. What appears to be a small amount of
damage on a harvested leaf is often more conspicuous following
curing.
Pathogen Description
Powdery mildew is caused by Erysiphe cichoracearum DC in the
class of fungi Pyrenomycetes, family Erysiphaceae. Erysiphe
cichoracearum produces both sexual and asexual spores. The asexual
spores, called conidia, are formed in chains (end-to-end) from the tips
of short conidiophores within the visible mold on the leaf surface.
Conidia are elliptical in shape, are single cells with thin walls and are
in the size range of 20 to 50 × 12 to 24 mm (Lucas, 1975). The sexual
spores are called ascospores and form in small sacs called asci which
are produced in a spherical structure called a perithecium. The
perithecia form within the mold on the leaf surface, usually toward the
end of the growing season. The black perithecia range in size between
80 and 140 mm and are easy to see with a hand lens. The perithecia
remain dormant until the next growing season and may survive for
several years.
Epidemiology
Powdery mildew epidemics can originate from either air-borne
conidia or from ascospores. In climates where tobacco or wild species
of Nicotiana survive throughout the year, conidia produced on these
plants serve as the primary source of the pathogen to infect new
tobacco plantings. In more northern climates with killing frosts and
freezing temperatures, the primary sources of the pathogen are
ascospores which are released from the

 Page 187
asci in the perithecia in the spring. Unlike most fungal leaf pathogens,
powdery mildew spores (ascospores and conidia) do not require free
water to germinate. The spores can germinate within 2 hours under
optimum humidity between 60 and 80%. The fungus grows more
readily on the older, fully expanded leaves. The grayish-white mold is
visible in about 7 days and conidia are produced in an additional 4
days. Conidia are released into the air in the late morning, reach a
peak by mid afternoon and germinate during the night. Warm daytime
temperatures with low to moderate humidity and cool nights with dew
formation favor spread and disease increase.
Management
Nicotiana digluta has been used as a source of resistance to powdery
mildew. In countries where powdery mildew is a serious problem,
breeding efforts to obtain resistant cultivars generally have been
successful. Fungicides can provide effective control of the disease, but
care must be exercised to avoid excessive residues on the harvested
leaf. Dinocap and benomyl have given good results, but benomyl
resistant strains of E. cichoracearum have been reported.
Experimental fungicides with high levels of activity against powdery
mildews are being developed and may offer improved disease
management in the future.
d
Wildfire and Angular Leaf Spot
Wildfire and angular leaf spot are bacterial diseases of tobacco which
have caused serious problems in the past, but are much less of a
problem today. These diseases have occurred in many tobacco
producing countries, especially in wet temperate climates. The
introduction of wildfire resistant cultivars in the 1950s has all but
eliminated wildfire, and the switch from outdoor seedbeds to
greenhouse seedling production is contributing to reductions in the
occurrence of angular leaf spot.
Symptoms
The bacteria which causes wildfire and angular leaf spot produce
numerous water-soaked leaf spots which range in diameter between 1
and 8 mm. The tissue in the spots dies quickly, dries and turns reddish
brown, brown or black. In wildfire, the spot is surrounded by a large
and conspicuous yellow halo which is lacking in angular leaf spot.
Infected seedlings can be killed by the wildfire disease, whereas
seedlings have not been known to be killed by angular leaf spot.
However, angular leaf spot infections on young expanding leaves can
cause severe leaf distortion. Both diseases can occur in the field.
Wildfire is now rare because of cultivar resistance, but at one time it
caused severe leaf damage. Angular leaf spot is occasionally a field
problem, especially following wind-blown rains. When severe
spotting occurs, the spots coalesce and give the leaf a ragged and torn
appearance. These diseases may be tentatively diagnosed by making a
glass slide mount of a leaf spot section in water and microscopically
observing streaming of bacteria from the infected tissue.
Pathogen Description
The bacteria that cause wildfire and angular leaf spot are in the
fluorescent group of phytopathogenic bacteria called pseudomonads.
They were once classified as separate species, but are currently placed
within the same species and pathovar designation, Pseudomonas
syringae pv. tabaci (Wolf & Foster) (Young, et al). Deall and Cole
(1986) have argued for separate pathovar status, i.e. P. syringae pv.
tabaci for the wildfire bacteria and P. syringae pv. angulata for the
angular leaf spot bacterium, because of differences in pathogenicity
and epidemiology. On the basis of morphology and standard
biochemical tests for identifying bacteria, the two organisms are
indistinguishable. The one characteristic in which they do differ is
their ability to produce tabtoxin, the toxin responsible for the yellow
halo in the wildfire disease. The angular leaf spot bacterium does not
produce tabtoxin. There are likely other differences since wildfire
resistant tobacco genotypes are not resistant to angular leaf spot.
Pseudomonas syringae pv. tabaci produces rodshaped cells, 0.5 × 2 to
2.5 mm in size, with one to six polar and bipolar flagella (Lucas,
1975). The bacteria are noncapsulate, nonsporeforming, nonacid-fast,
gram-negative and produce a fluorescent pigment in culture. When
grown on beef extract agar, colonies are white, slightly raised and
have a translucent edge with an opaque center (Lucas, 1975). Isolates
lose their ability to cause disease if repeatedly grown and transferred
in culture media. However, pathogenic isolates can be stored for long
periods in sterile distilled water and still maintain their ability to cause
disease.
Epidemiology
Like many foliar bacterial plant diseases, wet weather favors disease
development. When the tobacco leaves are wet, the bacteria will ooze
out of the leaf spots and will spread from leaf to leaf and plant to plant
by splashing rain or irrigation water. Infection is through

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stomates and wounds and is facilitated by wind-blown rains that force
water and bacteria into intercellular spaces within the leaf. Under
extreme wet conditions, the bacteria multiply and spread rapidly. Hot
and dry weather effectively stops these diseases.
Plant refuse from previously infected plants is a likely source of the
bacteria to begin new epidemics. Pseudomonas syringae pv. tabaci
has also been found on various weed species and the roots of crop
plants such as barley, rye and wheat. Seed has also been implicated as
a source of angular leaf spot outbreaks. In the past, the disease has
first appeared and been most severe in the seed bed. Certain cultural
practices in the seedbed can actually increase these diseases. Frequent
overhead watering can aid in the spread and increase of the bacteria
and clipping causes wounds which can aid in entry as well as spread
on the clipper. Further spread can occur if seedlings are pulled when
plants are wet. These diseases have generally been less of a problem
in the field, but have occasionally caused serious leaf damage and
loss.
Management
Seedbed sites should be rotated each year and should not be situated
where surface run-off from a previous crop can wash into the seedbed.
The increased use of float plant production in greenhouses, where
overhead watering is not used, should lead to a decrease of these
particular bacterial diseases. Use of resistant cultivars is the most
effective control for wildfire. Wildfire resistance is conferred by a
single dominate gene derived from N. longiflora. However, strains of
the wildfire bacterium that overcome this source of resistance have
been reported. Both bluestone-lime (Bordeaux mixture) and
streptomycin provide effective control when used in a preventive
program in the seedbed. Bactericides are not recommended or needed
in the field.
e
Hollow Stalk
Bacterial hollow stalk is one of three phases of bacterial diseases that
can affect tobacco from seedling to harvest. The bacteria that cause
hollow stalk also cause the seedling disease known as black leg and
the curing disease known as barn rot. These diseases occur wherever
tobacco is grown and harvested in warm moist climates.
Symptoms
Seedlings with black leg develop a soft watery rot of basal leaves and
stems. The affected tissue is slimy and appears light brown at first
then turns black upon drying. If the disease is present at the time of
pulling for transplanting, additional spread and increase may occur if
plants are held in containers overnight. In the field, the disease is
usually noticed following topping when affected plants appear to
suddenly wilt. Leaves droop and fall off the plant. The pith in the
main stem is at first slimy then turns black and the stalk becomes
hollow. Black streaks may be visible on the exterior of the stalk
extending into leaf petioles. Infection may be initiated in the cut or
break at topping, in which case the disease advances down the stalk.
Also, infection may be initiated in the cuts where leaves are primed at
the base of the plant, in which case the disease advances up the stalk.
The barn rot phase may develop in aircured tobacco in the curing barn
if the humidity is near saturation. Decaying leaves may slip off their
support strings or fall off the stalk of hung tobacco, which is
accompanied by the characteristic smell of soft rot. Mechanically
harvested tobacco may decay in the bulk bins especially if harvested
when wet and packed too tightly.
Pathogen Description
Black leg, hollow stalk and barn rot are caused by the common soft
rot bacterium Erwinia carotovora subsp. carotovora (Jones) (Bergey,
et al.). In addition, a similar bacterium, Erwinia chrysanthemi
(Burkholder, et al.) can cause barn rot. The soft rot bacteria are
rodshaped, in the size range 0.5 to 0.8 mm × 1.5 to 3.0 mm, and may
occur singly, in pairs or in chains, many of which possess from two to
eight flagella (Lucas, 1975). These bacteria are nonsporeforming,
noncapsulate, gram-negative and facultatively anaerobic. The soft rot
bacteria produce pectolytic and other enzymes which macerate and
soften plant tissues in advance of invasion into affected tissues.
Epidemiology
Erwinia catotoyota subsp. carotovora are common soil bacteria which
enter the plant through wounds. The bacteria can be disseminated on
soil particles, splash drops, insects and by air. On seedlings, the
bacteria usually infect leaves in contact with the soil and subsequently
invade the stem through the petiole, thus causing black leg. In the
field, the bacteria are common residents on leaf surfaces where they
do not normally cause a problem. However, if plants are pruned,
topped or suckered under wet conditions, and the bacteria come in
contact with the wound, they can readily enter the pith and cause
hollow stalk. Plants and leaves that

 Page 189
are harvested and handled when wet can result in the barn rot phase of
the disease. The presence of moisture is the most important factor in
the development of the three soft rot diseases on tobacco. Erwinia
carotovora subsp, carotovora grows over a wide temperature range (0
to 40°C) with optimal growth at warm temperatures (27 to 30°C).
Management
Moisture control and avoiding certain cultural practices when plants
are wet are important considerations in managing the three diseases.
No chemical controls or resistance are available. To reduce black leg
in the seedbed, covers should be removed early to reduce humidity,
and frequent overhead irrigation should be avoided. In the
greenhouse, humidity should be controlled through proper ventilation
and plants should be kept dry. To prevent hollow stalk, plants should
not be topped or suckered when plants are wet from dew or rain, or
rain is imminent. To prevent the barn rot phase, leaves and plants
should be dry when hung in the barn. If the tobacco is wet, heaters and
ventilation should be used to reduce the humidity. Mechanically
harvested tobacco should not be harvested when plants are wet nor
packed too tightly in bulk bins.
f
Black Shank
Black shank is a root and stem rot disease of all types of cultivated
tobacco (Lucas, 1975). Major losses occur each year in flue-cured and
burley tobaccos, but other tobacco types also may suffer severe losses.
Losses can occur during all stages of plant development and can reach
100% in highly infested fields under optimum disease conditions
(Lucas, 1975). The disease was originally described by Van Breda de
Haan in Indonesia in 1896, but it is not known where the fungus
originated. Since the first report, the fungus has spread to tobacco
growing regions worldwide; however, the disease does not occur in all
countries where tobacco is produced (Delon, et al., 1987). Since
tobacco appears to be the only natural host for the fungus, spread
probably occurred when tobacco and tobacco products were moved
between countries.
Symptoms
The black shank fungus affects the roots, basal stem region and leaves
of the tobacco plant, but symptoms vary with plant age, variety of
tobacco grown and weather conditions (Lucas, 1975; Shew & Lucas,
1991). Young seedlings are very susceptible, and typical damping-off
may develop in the seed bed or on greenhouse-produced seedlings
during periods of wet, warm weather. The initial symptom on field
plants is wilting of the leaves during the middle of the day. In general
all leaves wilt, which is in contrast to the unilateral or one-sided
wilting caused by vascular wilt pathogens. Leaves begin to turn
yellow and hang down the stalk over the next few days to weeks, and
as the disease progresses, the pathogen grows into and up the stem of
the plant. In its final stages, aboveground parts of the stem, or shank,
of the plant will turn black and hence the name, black shank.
When the stem of a diseased plant is split longitudinally, the pith
appears dry, is brown to black in color and is often separated into
plate-like disks up to the top of the stem lesion. Although this
symptom is characteristic of black shank, the disking symptom alone
does not confirm that the disease is black shank. Other diseases, such
as sore shin, also may cause disking of the pith under some
conditions, and the disking caused by lightning injury is not
discolored and extends well above any necrotic stem lesions produced
(Shew & Lucas, 1991).
During periods of rainy weather, lower leaves of tobacco also may
become infected as soil or spores of the fungus are splashed onto the
leaves. Leaf spots begin as small water-soaked spots, but they expand
rapidly, turn brown and necrotic and develop into circular lesions. The
fungus can completely destroy the leaf and then grow down the
petiole of the leaf into the stem and cause typical black shank
symptoms.
Pathogen Description
Van Breda de Haan named the fungus that causes black shank
Phytophthora nicotianae. Tucker renamed the fungus P. parasitica
Dast. var. nicotianae (B. de Haan) Tucker, whereas Waterhouse
concluded that the correct name of the black shank pathogen is P.
nicotianae B. de Haan var. nicotianae Waterhouse. Currently, both
names are used to describe the organism (Shew & Lucas, 1991).
The fungus can be easily grown in pure culture in the lab, but culture
morphology varies with isolate and growth media (Lucas, 1975; Shew
& Lucas, 1991). The fungus produces four spore stages: sporangia,
zoospores, chlamydospores and oospores (Goodins & Lucas, 1959;
Lucas, 1975). Sporangia are produced in wet soils and on roots and
germinate to produce hyphae or zoospores. Between 5 and 30
zoospores are produced per sporangium and can swim through the soil
for many hours or until they contact nutrients, toxic ions or

 Page 190
general toxicants such as fungicides. Zoospores infect the root tips of
the tobacco plant. Chlamydospores frequently develop in great
abundance in diseased tissue and allow the fungus to survive for long
periods in the soil. They germinate to produce hyphae or sporangia
when a host is present. Although oospores are sometimes produced,
they are not important in the life cycle of the fungus or the
epidemiology of the disease.
There are at least four known races of the pathogen. Race 0 is
predominant, but other races have been reported from specific
geographic locations (Prinsloo & Pauer, 1973; Shew & Lucas, 1991).
Races are identified by the resistance genes which are used in
cultivars grown in a particular area.
The fungus can be recovered or detected in soil or plant material by
baiting procedures, selective agar media and immunoassay
(Kannwischer & Mitchell, 1978; Shew & Lucas, 1991). Selective
media are used for the detection and quantification of P. parasitica
var. nicotianae which has helped in diagnosis and in understanding
the effectiveness of management practices such as fungicides, crop
rotation and resistance.
Epidemiology
Black shank is a warm-weather disease (Lucas, 1975). Soil
temperatures above 20°C are required for significant levels of
infection to occur, although infection can occur at 16°C. This may
partially explain the absence or poor development of the disease in
seedbeds in some locations. Lesion expansion is greatest at 22° to
28°C, and high soil moisture conditions enhance disease. Little or no
infection occurs in soil that is not saturated for at least brief periods of
time (Shew, 1983).
Soil factors affect pathogen activity and development of black shank
(Wills & Moore, 1969; Kincaid, et al., 1972; Lucas, 1975; Sidebottom
& Shew, 1985). Disease occurs in heavy and light soils, and soil
organic matter content has little effect on disease incidence. Black
shank is generally less severe in poorly drained soils, probably
because such soils are usually more acid and flooded conditions are
not optimum for fungus survival. Soil series and soil texture affect
both reproduction of the fungus and root infection (Lucas, 1975;
Sidebottom & Shew, 1985).
Black shank is most severe when soil pH is between 6.0 and 7.0.
Levels of calcium and magnesium in these soils are thought to
enhance leaf quality, but they are also associated with increased levels
of disease. These increased losses to black shank may eliminate any
benefit gained by the additional calcium amendment, as they may
increase the need for fungicide applications. The level of soil
aluminum affects disease severity; as aluminum goes up, disease
severity is decreased.
Low levels of overwintering inoculum are sufficient to initiate
epidemics of the disease because black shank is a polycyclic disease
(Ferrin & Mitchell, 1986; Shew, 1987). This means that the pathogen
is capable of going through many generations within a single growing
season. New sporangia and chlamydospores develop on roots within
48 hours of infection and this secondary inoculum spreads to healthy
plants and new cycles of infection occur (Shew, 1987). The pathogen
is easily spread on infected transplants, in water, soil and on tobacco
debris. Infected transplants may not show any symptoms, but under
favorable conditions the disease will continue to develop after
transplanting. Rain water transports infested soil and fungus spores
along rows and from infested fields into uninfested areas of the field
and into drainage ponds, creeks and rivers. The fungus also can be
present in stalks of infected plants, so these stalks should not be
placed on seedbed sites or fields after tobacco is stripped.
The severity of black shank is greatly increased in the presence of root
knot nematodes (Lucas, 1975). Nematodes provide a means of entry
into roots for the fungus. Black shank resistant cultivars become
susceptible when infected by root knot nematodes.
Losses to black shank primarily are due to plant death, especially in
tobacco types such as burley which have little resistance. In resistant
cultivars of flue-cured tobacco, leaves on affected plants ripen
prematurely, and many leaves can be saved if they are harvested as
they begin to yellow or wilt. These leaves may cure poorly compared
to leaves that ripen at a normal rate. If not harvested quickly, leaves
on affected plants turn brown and shrivel and are not marketable.
Management
Management of black shank requires the use of resistant cultivars,
crop rotation, nematode control and, in some fields, either soil
fumigants or fungicides. The fungicide metalaxyl has been effective in
reducing losses to black shank, especially when used in conjunction
with other management strategies. Cultivars vary significantly in their
level of resistance, so selection is critical. Susceptible cultivars are
still widely used in some types of tobacco, and rotation with nonhosts
or resistant cultivars is necessary to prevent build-up of inoculum in
these cases. It is critical to maintain soil pH below 6.0 if the field has
a history of black shank. High pH levels make disease management
very difficult by favoring rapid growth and reproduc-

 Page 191
tion by the fungus which reduces efficacy of fungicides and
effectiveness of host resistance.
g
Black Root Rot
Black root rot occurs in most tobacco growing regions of the world
(Delon, et al., 1987), but it is most severe on tobacco grown in cooler
climates (e.g. Europe, Canada, northern USA). The pathogen attacks
more than 100 species in 33 families of plants. The first reports of
black root rot on tobacco were from the United States and Italy around
1880 (Lucas, 1975). The fungus has a worldwide distribution and
likely attacked other plants prior to the introduction of tobacco into
various countries; black root rot is not an important disease in
countries where tobacco originated. Losses occur in the seedbed and
field, although seedbed losses are greatly reduced or eliminated by
fumigation treatments. Annual losses of 5% or higher are common in
some countries, especially where no host resistance has been
incorporated into varieties.
Symptoms
Roots infected with the black root rot fungus develop black lesions
that may coalesce to form large necrotic areas along the entire length
of the root (Shew & Lucas, 1991). Small roots are rotted through
completely, resulting in a much reduced root system and the presence
of numerous blackened root tips upon removal of plants from the soil.
Lesions on large roots are black, scaly and slightly sunken.
Tobacco roots are susceptible throughout the growing season, but in
many countries black root rot is considered a seedling disease.
Seedlings with only a few infected roots show no aboveground
symptoms, but severely infected seedlings are stunted and turn pale
green to yellow and may even be killed. In the field, the first
indication that black root rot is present is an uneven growth among
plants. On hot days, leaves of diseased plants wilt more quickly than
those of healthy plants because of their reduced ability to take up
water. These symptoms are common to many root diseases, so roots
must be examined carefully before an accurate diagnosis can be made.
In addition to the characteristic black lesions, examination of diseased
tissue with a microscope will reveal the presence of the characteristic
spores of the fungus.
Pathogen Description
Taxonomy of the black root rot fungus has been the subject of
considerable debate (Shew & Lucas, 1991) and at present, both
Thielaviopsis basicola (Berk. & Broome) Ferraris and Chalara
elegans Nag Raj & Kendrick are used to identify the black root rot
fungus. Thielaviopsis basicola is the correct name for chamydospore
stage of the fungus and will be used in this chapter. The C. elegans
name refers only to the endocondial state of the organism and is
referred to as a synanamorph of T. basicola.
Isolates of T. basicola are easily grown under laboratory conditions.
The fungus produces two spore types, endoconidia and
chlamydospores. Both are produced in culture and on host tissue. The
fungus grows well on a variety of natural media including PDA and
carrot agar (4 to 8%) with optimum temperatures for growth in culture
between 22° and 30°C. Spore germination is rapid between 21° and
33°C. Optimum pH for growth in culture is between 4.0 and 6.2.
Some host specialization is present in T. basicola (Corbaz, 1985), but
only one race of T. basicola is known on tobacco.
Quantitative recovery of T. basicola from soil has been difficult, but a
selective medium developed by Specht and Griffin (1985) has been
very effective in recovery of T. basicola from soil and plant tissue.
Epidemiology
Black root rot severity depends primarily upon the prevailing soil
temperature, inoculum density of the pathogen, soil chemistry (pH
related factors) and level of host resistance (Lucas, 1975; Meyer &
Shew, 1990; Meyer & Shew, 1991). Endoconidia and chlamydospores
germinate and infect tobacco roots, and as the fungus colonizes the
root, small roots are killed and many root tips are lost. This loss
affects synthesis of many of the important chemical compounds in
tobacco, and results in a thin leaf of some reduced quality.
Inoculum levels present in soil vary with cropping history and soil
type. Inoculum density required to cause significant levels of root rot
varies with soil type and level of host resistance in the cultivar planted
(Meyer & Shew, 1990; Meyer & Shew, 1991). Inoculum of T.
basicola can survive in soil for several years, but populations drop
rapidly in the absence of a host. However, because of the wide host
range, careful selection of rotation crops is important.
Chlamydospores are the primary survival structures and are spread in
soil, water and on roots of transplants.
Black root rot is most severe in cool soils (Lucas, 1975). Soil
temperatures between 17° and 23°C are generally considered to be the
most favorable for dis-

 Page 192
ease development. Although these temperatures are not optimum for
growth of the fungus, growth of tobacco also is slowed at these
temperatures and black root rot develops. The disease is most severe
at temperatures unfavorable for host growth, and symptoms often
disappear as soil temperatures increase and root growth is rapid. The
fungus remains active on the roots of susceptible plants, and
secondary inoculum produced may contribute to disease development
the following growing season.
Soil acidity is of primary importance in controlling this disease. The
disease is usually not severe below pH 5.6, however this is not strictly
a pH effect since the fungus grows well in culture between pH 4.0 and
6.4 (Lucas, 1975). The mechanism of disease suppression in acid soils
has been related to the level of exchangeable aluminum present
(Meyer & Shew, 1991). The addition of calcium ions by liming with
calcium carbonate increases disease severity, while use of gypsum
does not enhance disease.
Management
A combination of practices is required to adequately manage black
root rot. Where possible, rotation should be used because it reduces
inoculum density of the pathogen. Leguminous cover crops should be
avoided as T. basicola populations may reach high levels on these
crops. Rye and hairy vetch appear to be effective cover crops as they
are not hosts or they suppress populations of T. basicola.
There are two known sources of resistance to T. basicola in tobacco
(Lucas, 1975). Resistance derived from N. tabacum is multigenic, and
cultivars with levels of partial resistance from low to high have been
developed. Single-gene resistance from N. debneyi provides complete
resistance to T. basicola. Because T. basicola is a variable organism,
development of races capable of overcoming the single gene
resistance is possible.
Some chemical controls are available for use against T. basicola.
Methyl bromide gives excellent control in the seed bed, and trizole
fungicides, the demethylating inhibitors (DMI), are active against the
fungus. Fumigation of field soils with chloropicrin or other general
soil sterilants also may be helpful, but local recommendations should
be followed regarding the use of chemicals for black root rot control.
Finally, soil should not be overlimed. The pH should be maintained
between 5.5 and 5.8 in infested fields. Higher pHs favor black root rot
more than the added benefit obtained for plant growth.
h
Sore Shin and Damping off
All tobacco types are susceptible to sore shin. The pathogen is present
in most tobacco areas worldwide (Delon, et al., 1987) because the
fungus has a very wide host range, yet losses rarely exceed 1% in a
given field. The disease can cause extensive damage to tobacco
seedlings, whether produced in the seedbed or greenhouse, so
management practices that limit development of sore shin are
essential in the production of healthy seedlings. The disease is seldom
important if healthy seedlings are used in transplanting.
Symptoms
The first symptom of sore shin is a small water-soaked area on the
stem near the soil line. This area rapidly becomes dark brown and
sunken, and it may enlarge until the stem is girdled (Lucas, 1975;
Shew & Lucas, 1991). Often, a tuft of mycelium is visible in the
center of the lesion. If infected seedlings are transplanted, small
lesions may continue to develop into a canker that extends up to the
lower leaves. This symptom may be confused with black shank since
disking of the pith may occur, however, characteristic mycelium of
Rhizoctonia will be present.
When small lesions are present, plants may continue to grow and
appear normal. However, the stem often will be very constricted at the
soil line and the plants easily break off or are blown over during wind
storms. The root system is not decayed, but it may be less extensive
than on healthy plants. Leaf quality is generally not affected, but
plants which blow are difficult to harvest.
Patbogen Description
Sore shin is caused by the soilborne fungus Rhizoctonia solani Kuhn
(teleomorph Thanatephorus cucurneris (Frank) Donk). Not all isolates
of R. solani from other host plants will cause damping off and sore
shin of tobacco; sore-shin isolates that have been characterized belong
to AG-1, AG 3 and AG-4 (compare: only AG-3 causes target spot)
and are very diverse in their morphology (Ogoshi, 1987).
Epidemiology
Rhizoctonia solani survives in soil as hyphae in colonized organic
matter and as sclerotia. The fungus exists in soil indefinitely in the
absence of antagonists or suppressive environments. Optimum
temperatures for germination and growth are usually between 24° and

 Page 193
28°C, but rapid growth occurs between 20° and 30°C (Parmeter,
1970).
Germination of sclerotia is stimulated by host exudates. Hyphae that
contact tobacco stems branch profusely prior to infection of stem
tissue. Infected seedlings may be killed (damped off) or the disease
may be limited to discrete lesions on the stem. New inoculum is
produced on, or in, host tissue and the cycle is repeated when new
substrates become available. The fungus tolerates both acid and
alkaline soils. Losses to the disease are generally limited to seedlings,
so the only effect on tobacco production is through loss of plants. On
large plants, losses are limited to plants that blow over.
Management
Fumigation of seedbed sites reduces the incidence of sore shin on
seedlings. In the field, preventing injuries to the transplants reduces
infections by R. solani. Bruising of the stem during handling and
fertilizer injury are the most common types of injury that lead to
development of sore shin.
In the greenhouse, extreme care should be exercised to prevent the
introduction of inoculum to seedling trays. All trays and media should
be sterilized prior to use with either steam or methyl bromide. Surface
disinfestants have not proven successful in sterilization of trays. No
chemicals are registered for management of R. solani in the
greenhouse, but exemptions for the use of Rovral (iprodione) have
been obtained in some states in the USA.
i
Target Spot
A leaf-spot disease of tobacco caused by basidiospores of
Thanatephorus cucumeris (Frank) Donk was reported from Brazil in
1948 (Costa, 1948) and from Costa Rica in 1973 (Vargas, 1973). In
the United States, the disease was first observed in North Carolina in
1983 (Shew & Main, 1985), and it has been observed or reported from
many other countries including South Africa (Meyer, et al., 1990) and
Zimbabwe (Shew & Melton, 1995). The disease probably occurs in
other countries with mild, humid climates. The disease may easily be
confused with other leaf spots and appears to be a new threat to
tobacco worldwide. Losses are usually negligible, but during extended
periods of favorable environmental conditions they may exceed 50%
in a given field. For example, severe epidemics were reported from
North Carolina in 1989 and 1995 and resulted in total losses of over
$50 million US.
Symptoms
Symptoms begin as small circular water-soaked spots about 2 to 3 mm
in diameter. On seedlings, these initial lesions are net-like in
appearance when leaves are held up to a light source. On field plants,
these small spots may continue to expand, and under conditions of
high relative humidity and moderate temperature, lesions enlarge
rapidly, becoming light green and almost transparent with irregular
margins and chlorotic halos, and may completely destroy fully
expanded leaves. In less humid conditions, lesions expand more
slowly and often develop a pattern of concentric rings. This symptom
makes target spot somewhat difficult to distinguish from Ahernaria
brown spot (Shew & Melton, 1995). Necrotic tissue becomes brittle
and often drops out leaving a shot-hole effect in the leaf, and a
chlorotic halo often is present around expanding lesions.
Pathogen Description
Target spot (Rhizoctonia leaf spot) is caused by Thanatephorus
cucumeris Donk (anamorph Rhizoctonia solani Kuhn). Isolates of T.
cucumeris are similar in morphology, and all belong to anastomosis
group 3(AG-3) (Shew & Melton, 1995). Optimum temperatures for
growth are between 24° and 28°C, maximum 36°C. Basidiospores are
produced readily on soil and plants in favorable environments.
Isolates of T. cucumeris that cause leaf spot also may cause damping
off and sore shin under certain environmental conditions; however,
most sore shin isolates of R. solani are of a different anastomosis
grouping, AG-1 or AG-4, and do not cause leaf spot.
Epidemiology
Environmental conditions conducive to development of target spot of
tobacco include extended periods of high relative humidity or leaf
wetness and moderate temperatures (20° to 30°C). Also, these
conditions are necessary for abundant production of basidiospores,
infection and colonization of host tissues. When conditions are
unfavorable for basidiospore production (low moisture), leaf spot
isolates of T. cucumeris may cause damping off and sore shin of
tobacco seedlings.
Secondary inoculum production is important in leaf spot development.
New basidiospores are produced on leaves, other plant parts or on leaf
tissue that has fallen to the soil and are readily dispersed by air
currents to healthy tobacco tissue. Under optimum conditions, 16 to
20 days are required for completion of the disease cycle (Shew &
Main, 1990).

 Page 194
In an extensive host range study (Shew and Melton, 1995)
basidiospores of the tobacco leaf spot isolates of T. cucumeris
attacked only tobacco, sugar beet and egg plant. Only flue-cured and
burley type tobaccos were tested, but other tobacco types are probably
susceptible. Losses to target spot are through loss of leaf tissue.
Affected leaves may be ragged and somewhat thin upon curing.
Management
Sensitivity to fungicides is similar to that of sore shin isolates of R.
solani and the fungicide Rovral has been used in the US in
greenhouses and, in some cases, in the field to slow epidemics of the
disease. Sanitation practices should be used to prevent the
introduction of inoculum into seedling production beds or trays used
in greenhouse production of seedlings. Practices that limit wetting of
foliage (spacing, ventilation and flood irrigation in greenhouses)
reduce the incidence and severity of the disease. No resistance has
been identified.
j
Fusarium Wilt
Fusarium wilt of tobacco occurs in many countries (Lucas, 1975;
Delon, et al., 1987; Shew & Lucas, 1991). In the United States it is
the most destructive disease on broad leaf tobacco in Connecticut
(LaMondia & Taylor, 1987), and although the disease is widely
scattered in the flue-cured and burley production areas of the United
States, it seldom causes serious losses. It is a serious pathogen of
tobacco in many countries (Delon, et al., 1987).
Symptoms
The most conspicuous symptom is yellowing and bronzing of leaves
on one side of the plant. Wilting is not conspicuous at this stage of
disease. Leaves on the affected side of the plant also are dwarfed and
often curved due to uneven growth of the mid-rib. In addition, the tip
of the plant is often drawn over toward the diseased side, giving rise
to the common name 'draw-stalk' or 'crook-neck'. All the vascular
tissue on the affected side of the plant becomes a uniform medium-
brown as opposed to the black streaking of the vascular system caused
by the bacterial wilt pathogen. Sections of affected roots and mid-ribs
of affected leaves also show discoloration of the vascular system. In
typical cases, Fusarium wilt results in a dry rot of the stem in contrast
to the wet or slimy rot caused by the bacterial wilt pathogen.
The fungus often enters a single root and then spreads up that side of
the plant in the vascular tissue, resulting in loss of only a few leaves
on the plant. In other cases, all roots on one side of the plant may be
attacked. This severe infection causes severe stunting or death of
plants, resulting in severe loss of yield. Presence of the fungus and
brown discoloration of the wood give positive identification of the
disease.
Pathogen Description
Fusarium wilt of tobacco is caused by Fusarium oxysporum
(Schlecht) Wr. f. sp. nicotianae Johnson (Nelson, et al., 1981). There
is a close relationship among isolates of F. oxysporum from tobacco,
sweet potato and cotton, so rotation with these crops should be
avoided if the disease is present. Only one race of the tobacco
pathogen is known.
The fungus is easily cultured in the laboratory on standard growth
media and produces three spore stages: microconidia, macroconidia
and chlamydospores. No sexual spores are produced. The optimum
temperature for growth is 28° to 30°C, and the fungus grows over a
wide pH range. The fungus is notoriously variable, with loss of
virulence common in old cultures. Cultures should be stored as spores
in sand culture or frequently reisolated from inoculated plants to
maintain virulence.
Epidemiology
The pathogen can survive in soil for very long periods of time without
tobacco present. Dormant chlamydospores can survive as long as 10
years in soil. Chlamydospores germinate in response to host. The
fungus usually enters roots through wounds made by man, nematodes,
etc. and grows into the vascular system. After the disease becomes
established the vascular system turns brown as the result of enzymatic
attack on the vascular tissues, and symptoms appear within a week or
10 days of infection. Leaves of affected plants have a decreased sugar
content with an increase in resins and waxes. The disease also
adversely affects leaf quality.
Fusarium wilt is a warm weather disease; little damage occurs in cool
soils. Sandy-loam soils appear to be most favorable for disease
development, perhaps because they are also favorable for root knot
nematodes. Soil pH appears to be a minor factor as the disease occurs
in acid, neutral or alkaline soils, although the disease may be favored
in acid soils in some locations. Any moisture shortage sufficiently
severe to reduce the vegetative vigor of the host also will reduce
disease development.

 Page 195
Fusarium wilt is often associated with and accentuated by root knot
and tobacco cyst nematodes. The fungus is a wound parasite and
nematode-feeding punctures provide suitable infection courts for the
fungus. Also, nematodes alter host physiology, making it more
suitable for fungus growth. An interaction of root knot and fusarium
wilt with Alternaria brown spot has been reported.
Management
The use of resistant cultivars is the principal means of controlling
fusarium wilt, but resistance in tobacco to Fusarium wilt is not
complete. In conjunction with resistant cultivars, soil fumigation with
an acceptable nematicide will often prove beneficial in those fields
heavily infested with nematodes.
In areas where Fusarium wilt is a problem, tobacco should not be
grown in rotation with sweet potatoes, for both crops are susceptible
to the same fungus strains. Flue-cured tobacco may be grown safely in
sequence with cotton, but burley and dark tobacco should not be
grown in rotation with cotton.
k
Collar Rot
Collar rot has generally been of minor importance in tobacco
production (Delon, et al., 1987), but as greenhouse production of
transplants has increased, so has the importance of collar rot. The
disease is included in this treatment to call attention to the potential of
a new threat to tobacco in areas where the disease has not occurred
previously. The disease is generally considered to be important in
areas with cool climates, similar to those conducive to black root rot.
Symptoms
Seedlings may be attacked at any time, but in general the disease is
much more severe after leaves of seedlings make contact and form a
canopy (Shew and Lucas, 1991). Infection is first noticed as water-
soaked lesions on lower leaves followed by the development of a
water-soaked stem lesion. Affected stems develop a brown soft rot
which spreads into the leaves and rapid destruction of seedlings
occurs. During periods of high relative humidity, white, cottony
mycelium grows over the rotted plants, and black sclerotia develop in
the pith and on the surface of stems. Field symptoms result in leaves
that are of poor quality, similar to those observed with other diseases
that cause wilting and yellowing.
Pathogen Description
Collar rot is caused by Sclerotinia sclerotiorum (Lib.) Dby (Shew &
Lucas, 1991). The fungus survives as black, hard, oblong sclerotia (5
to 10 mm × 3 to 6 mm) in plant tissue or in soil. Sclerotia can survive
for many years in soil in the absence of a host. Sclerotia germinate by
production of vegetative hyphae that can infect host tissue or by
production of a specialized structure known as an apothecium.
Apothecia are small yellow to orange, cup-shaped structures that
release ascospores. The ascospores are wind dispersed and when they
land on plants they germinate in a thin film of water on the plant
surface, especially in the presence of leaf debris (for example, after
clipping of foliage). Periods of cool wet weather favor disease
development and spread of the fungus between closely spaced
seedlings in the field or greenhouse.
Management
Fumigation of seedbed sites will eliminate inoculum of the fungus in
the bed, but ascospores of the fungus may blow into the bed from
surrounding areas. In the greenhouse, sanitation to remove all infected
plant material or infested soil is critical to prevent disease
development. Diseased seedlings should not be discarded close to the
greenhouse as sclerotia which form on the tissue may produce
inoculum for the following growing season, which can initiate new
epidemics of the disease. Adequate ventilation and avoiding dense
plant stands will aid in maintaining an environment unfavorable for
the germination and growth of the fungus. Fungicides may be
effective in suppressing disease, but adequate coverage is necessary.
l
Bacterial Wilt
Bacterial wilt, also known as Granville wilt, first attracted attention in
Granville County, North Carolina, where it caused serious damage on
tobacco farms in about 1880 (Lucas, 1975). Bacterial wilt has since
been reported from tobacco growing regions in tropical, semitropical
and warm-temperate zones of the world (Buddenhagen, 1985;
Englebrecht & Prinsloo, 1985; Akiew & Trevorrow, 1994; Hayward
& Hartman, 1994) and is considered particularly serious in Asia and
North, South and Central America (Akiew & Trevorrow, 1994). The
bacterium has a very wide host range, but host specialization limits
the range for a given strain of the bacterium (Hayward & Hartman,
1994).

 Page 196
Symptoms
Bacterial wilt is characterized by wilting, stunting and yellowing of
the foliage and may occur at any growth stage of the plant (Lucas,
1975). On young, succulent, susceptible plants, initial symptoms are
wilting of one or two leaves during the hot part of the day, frequently
followed by recovery in the evening and early hours of the morning.
Often leaves on one side of the plant or even half a single leaf become
wilted. This unilateral wilting is a characteristic symptom of bacterial
wilt. If the disease develops rapidly the leaves wilt without change in
color. However, if wilt occurs gradually, affected leaves become light
green and progressively turn yellow, and frequently necrotic areas
appear between the veins and at leaf margins. In hot, dry weather,
wilted leaves are often scalded.
A cross section of the stem of a diseased plant reveals a tan to yellow-
brown discoloration of the vascular tissue. As the disease progresses,
large portions of the pith and cortex become deep-brown to black. In
contrast to fusarium wilt, the vascular tissue of plants with bacterial
wilt has dark brown to black streaks (pencil markings) and not a
uniform medium-brown discoloration as seen in plants with fusarium
wilt. Finally, as the pith decays the stem at the base, the plant becomes
hollow and the vascular tissue turns dark brown to black. Plants often
develop stem lesions similar to those observed with black shank
(Shew & Lucas, 1991).
A reliable diagnostic test for bacterial wilt is the observation of
masses of bacterial cells in vessels. If the stem of a wilted plant is cut
in cross section and left in a humid environment for a few hours,
dirty-white glistening droplets of a viscous ooze visible with the
naked eye are exuded from the vascular tissue. If a thin longitudinal
section of one of the blackened streaks in the vascular tissue is
removed, placed in a drop of water and examined with a microscope,
bacterial slime will be observed almost immediately. The presence of
ooze can also be demonstrated by placing a longitudinal section of a
diseased stem in a container of water. In a few minutes, fine milky-
white strands composed of masses of bacterial cells and slime stream
from the vascular tissue.
Pathogen Description
Ralstonia solanacearum E.F. Smith is a nonfluorescent,
nonsporeforming, Gram-negative rod (Hayward & Hartman, 1994).
Strains of R. solanacearum vary greatly in their ability to attack
tobacco, and strains from different countries are capable of attacking
different varieties of tobacco (Akiew & Trevorrow, 1994). Isolates
change rapidly when maintained on most laboratory media, so long
term storage should be done in sterile distilled water.
Epidemiology
Severity of bacterial wilt depends primarily on soil temperature and
moisture, the amount of pathogen present and the level of host
resistance. Ralstonia solanacearum penetrates tobacco roots through
wounds caused by the emergence of lateral roots or by soil organisms
such as nematodes. Once the pathogen has penetrated, time required
for symptom expression is variable and depends on environmental
conditions and host resistance. Inoculum is readily spread in soil,
water or transplant seedlings.
Temperature plays an important role in the geographic distribution of
the pathogen and in development of the disease. Bacterial wilt
develops most rapidly at 30° to 35°C. High soil moisture favors
infection because it increases contact of host roots by the bacterium
and enhances survival of the pathogen. Wounding decreases the
number of bacteria needed to infect roots of tobacco.
Losses to bacterial wilt may be in total yield, as death may reach
100% in some fields, or in reduction in quality of leaf from infected
plants.
Management
There is no single practice that will provide complete control of
bacterial wilt. Therefore, an effective management program should
include crop rotation, nematode management, chemical application,
resistant cultivars, proper cultivation and stalk and root destruction at
the end of harvest.
References
Akiew, E. & Trevorrow, P.R. (1994) Management of bacterial wilt of
tobacco. In: Bacterial Wilt: The Disease and its Causative Agent,
Pseudomonas solanacearum (eds A.C. Hayward & G.L. Hartman).
pp. 179-98. CAB International, Wallingford, UK.
Buddenhagen, I.W. (1985) Bacterial wilt revisited. In: Proc. Int.
Workshop Bact. Wilt Dis. Asia South Pac. (ed. G.J. Persley). Aust.
Cen. Int. Agric. Res. (ACIAR) Proc., 13, 12643.
Corbaz, R. (1985) Pathotypes et variations due pouvoir pathogene
chez Charlara elegans Nag Raj et Kendrick (= Thielaviopsis
basicola). Phytopath. Z., 113, 28999.
Costa, A.S. (1948) Mancha aureolada e requeima do fumo causadas
por Corticium solani. Biologico, 14, 11314.
Deall, M.W. & Cole, J.S. (1986) A comparative study of the

 Page 197
pathogenicity and epidemiology of strains of Pseudomonas
syringae pv. tabaci that cause wildfire and angular leaf spot disease
of tobacco in Zimbabwe. Plant Pathol., 35, 7481.
Delon, R., Hill, D., Papenfus, H.D. & Tancogne, J. (1987) Survey of
Pests and Diseases of Tobacco and Chemicals used. Part A. Diseases
and Pests. Agron. Phytopath. Group, CORESTA, Paris.
Engelbrecht, M.C. & Prinsloo, G.C. (1985) Pseudomonas
solanacearum on tobacco in South Africa. Phytophylactica, 17, 1712.
Ferrin, D.M. & Mitchell, D.J. (1986) Influence of initial density and
distribution of inoculum on the epidemiology of tobacco black shank.
Phytopathol., 76, 2439.
Gooding, G.V. & Lucas, G.B. (1959) Factors influencing sporangia
formation and zoospore activity in Phytophthora parasitica var.
nicotianae. Phytopathol., 49, 27781.
Hayward, A.C. & Hartman, G.L. (eds) (1994) Bacterial Wilt: The
Disease and its Causative Agent, Pseudomonas solanacearum. CAB
International, Wallingford, UK.
Kannwischer, M.E. & Mitchell, D.J. (1978) The influence of a
fungicide on the epidemiology of black shank of tobacco.
Phytopathol., 68, 176065.
Kincaid, R.R., Martin, F.G. & Rhoads, F.M. (1972) Regressions of
tobacco black=shank index on soil calcium. Phytopathol., 62, 302.
LaMondia, J.A. & Taylor, G.S. (1987) Influence of the tobaccocyst
nematode (Globodera tabacum) on Fusarium wilt of Connecticut
broadleaf tobacco. Plant Dis., 71, 112932.
Lucas, G.B. (1975) Diseases of Tobacco, 3rd edn. Biological
Consulting Assoc, Raleigh.
Meyer, J. & Shew, H.D. (1990) Field development of black root rot on
burley tobacco as influenced by inoculum density, host resistance and
soil chemistry. Plant Dis., 74, 6015.
Meyer, J. & Shew, H.D. (1991) Soils suppressive to black root rot of
burley tobacco, caused by Thielaviopsis basicola. Phytopathol., 81,
94654.
Meyer, J.C., Van Wyk, R.J. & Phillips, A.J.L. (1990) Rhizoctonia leaf
spot in South Africa. Plant Pathol., 39, 206207.
Nelson, P.E., Toussoun, T.A. & Cook, R.J. (1981) Fusarium:
Diseases, Biology, and Taxonomy. The Pennsylvania State University
Press, University Park.
Ogoshi, A. (1987) Ecology and pathogenicity of anastomosis and
intraspecific groups of Rhizoctonia solani Kuhn. Ann. Rev.
Phytopathol., 25, 12543.
Parmeter, J.R., Jr (ed.) (1970) Rhizoctonia solani: Biology and
Pathology. University of California Press, Berkeley.
Prinsloo, G.C. & Pauer, G.D.C. (1973) Die identifikasie van rasse van
Phytophthora nicotianae (B. de Haan) nicotianae wat in Suid Afrika
voorkom. Phytophylactica, 6, 21720.
Shew, H.D. (1983) Effects of soil matrix potential on infection of
tobacco by Phytophthora parasitica var. nicotianae. Phytopathol., 73,
116063.
Shew, H.D. (1987) Effect of host resistance on spread of
Phytophthora parasitica var. nicotianae and subsequent development
of tobacco black shank under field conditions. Phytopathol., 77,
109093.
Shew, H.D. & Lucas, G.B. (1991) Compendium of Tobacco Diseases.
APS Press, St Paul.
Shew, H.D. & Main, C.E. (1985) Rhizoctonia leaf spot of fluecured
tobacco in North Carolina. Plant Dis., 69, 901903.
Shew, H.D. & Main, C.E. (1990) Infection and development of target
spot of fluecured tobacco caused by Thanatephorus cucumeris. Plant
Dis., 74, 100913.
Shew, H.D. & Melton, T.A. (1995) Target spot of tobacco. Plant Dis.,
79, 611.
Sidebottom, J.R. & Shew, H.D. (1985) Effect of soil type and soil
matric potential on infection of tobacco by Phytophthora parasitica
var. nicotianae. Phytopathol., 75, 143943.
Specht, L.P. & Griffin, G.J. (1985) A selective medium for
enumerating low populations of Thielaviopsis basicola in tobacco
field soil. Can. J. Plant Path., 7, 43841.
Vargas, E. (1973) Infeccion pot basidiospores de Thanatephorus
cucumeris, causante de una enfermedad foliar en tabaco. Turrialba,
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of tobacco. Phytopathol., 59, 34651.

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6B
Virus Diseases
D. Blancard and R. Delon
Institut du Tabac, Seita
Bergerac, France

B.W. Blair and T. Glover

Tobacco Research Board
Harare, Zimbabwe
Introduction
Tobacco is a host for numerous viruses, and significant economic crop
losses result each year from virus infection of this economically
important crop. Indeed, a recent survey of tobacco pests and diseases
by a CORESTA task force (Delon, et al., 1993) revealed that one
virus, (tobacco mosaic virus (TMV), was always an important
problem in many tobaccoproducing countries.
Virus-related diseases have long been a major concern for tobacco
growers because of the lack of efficient methods of disease control.
Tobacco viruses may damage leaves and stems and cause significant
economic losses by reducing yield and quality of the cured tobacco.
Losses may be catastrophic locally, or they may be mild and relatively
insignificant. The severity of virus diseases varies with the country,
locality, tobacco variety, climatic conditions and severity of an
individual virus strain. At many locations, however, virus diseases
occur in tobacco and cause losses ranging from small to moderate.
The cumulative effects of these endemic virus diseases are substantial
losses to growers and poorer quality tobacco for the manufacturers.
Symptoms Caused by Viruses
The most common, and sometimes the only, symptom of a virus
infection is a reduced growth rate of the plant. This may result in
varying degrees of dwarfing or stunting of the entire plant. Visible
symptoms are often the most intense in the early stages of tobacco
development. Many viruses cause a light and dark green mosaic or
mottling symptoms, slight or severe puckering of leaves, distortion of
stem or leaves or tissue proliferation (enation). Ring spots and rings
with lines or 'oak leaf' patterns are other types of leaf symptoms
commonly seen with many virus infections. The type and severity of
symptoms also can be greatly affected by environmental conditions,
especially temperature and light intensity. Finally, even if harvesting is
possible, the quality of virus-infected leaves may be greatly reduced.
Morphology and Structure
Basically, the plant virus particle consists of an infectious nucleic
acid, referred to as the viral genome, that is enclosed within a
protective protein coat that is sometimes named the capside protein.
The genome which carries the genetic information necessary for the
virus's replication is composed of ribonucleic acid (RNA) in most
groups of plant viruses but consists of deoxyribonucleic acid (DNA)
in members of the caulimovirus and geminivirus groups. The RNA or
DNA may be single or double stranded. As an example, Fig. 6.1
shows the structure of TMV as rod-shaped particles with the protein
coat and RNA genome surrounding an axial hole. The main
characteristics of tobacco viruses are described in Table 6.1.
Identification Methods
The proper identification of the virus infecting the tobacco plant is
key to designing any possible control strategy or to studying the
epidemiology of the virus. Before the development of serological
techniques, laborious and time consuming assays such as trans-


 Page 199

Fig. 6.1
An electron micrograph of tobacco mosaic virus
(TMV) particles shadowed with vapour of heavy
metal. Bar scale = 100 nm. (Courtesy of René ROHR.)
mission to indicator hosts were used. Some of these more
conventional techniques still have a place in the diagnosis of viral
pathogens, but generally, the identification of viruses combines
different techniques such as symptomology and bioassays with
serology tests.
a
Disease Symptoms
Information needed to identify a virus differs from one virus group to
another. An accurate description of symptoms helps greatly to identify
the causal virus. However, a thorough description of symptoms,
mosaic, necrosis, etc. is seldom sufficient to identify an unknown
virus precisely.
b
Inclusion Bodies
Many plant viruses induce inclusions in host cells, and the inclusions
are more or less specific to each virus. Coupled with light microscopy,
the staining of inclusion bodies allows the identification of many
viruses based on the morphology and color reaction of the inclusion
bodies.
c
Bioassays with Indicator Hosts
In the absence of an electron microscope, screening or indexing plant
material for viruses is often required. It involves tests for one or more
well recognized viruses that are known to occur in the plant species
concerned. This works well with sap transmissible viruses. The test
plants include: Chenopodium amaranticolor, Cucumis sativus, Curbita
pepo, Datura stamonium, etc. and different Nicotiana species.
d
Immuno Tests
Immunosera containing virus-specific antibodies are commonly used
to detect viruses in immunodiffusion tests, immunoenzymology or
ELISA (enzyme linked immunosorbant assays). ELISA, used
routinely in virus identification, is a very sensitive and quantitative
method. There are numerous citations that provide detailed procedures
for the different ELISA methods that are available. Monoclonal
antibodies can be used to differentiate among strains of the same
virus.
e
Molecular Techniques
The technique of nucleic acid hybridization is a relatively recent
development and it can be adapted for the detection and identification
of viruses. In this method, specific probes bind or hybridize only to
the nucleic acid that is unique for a particular virus. Probes with
radioactive labels have been developed for many viral plant
pathogens. Commercial development of nonradioactive DNA or RNA
probes for nucleic acid hybridization assays is starting to be
successful.
Another molecular tool, Polymerase Chain Reaction (PCR), which is
used to amplify discrete regions of nucleotides, will also find a place
in the diagnostic methods used in modern laboratories for DNA and
RNA viruses.
Infection Process
Plant viruses enter plant cells only through wounds made
mechanically or by vectors. After entry, the nucleic acid (RNA) of the
virus is first freed from the protein coat, a process called
decapsidation. The next step is the multiplication of the virus in the
host cell, or virus replication.
For infection of a plant by a virus to take place, the virus must move
from one cell to another and must multiply in most cells it enters. This
movement of a virus from cell to cell is made through the
plasmodesmata. The mechanisms of transport into vascular tissue are
not well known, but a large number of viruses are known to be rapidly
transported through the

Table 6.1 Characteristics of the major tobacco viruses.
Common AbbreviationGroup CharacteristicsMode of Occurrence
name transmission
Tobacco TMV TobamorvirusRigid rod 300 Mechanically Worldwide
mosaic × 18 nm and contact
virus
Cucumber CMV Cucumovirus Icosahedral Insects Worldwide
mosaic particles Æ 28 (nonpersistent)
virus nm and
mechanically
Potato PVY Potyvirus Flexuous rod Insects Worldwide
virus Y 730 × 11 nm (nonpersistent)
and
mechanically
Tobacco TEV Potyvirus Flexuous rod Insects Principally
etch virus 730 × 1213 (nonpersistent),in North,
nm mechanically, Central and
cuscuta South
America,
South Africa
and Far East
Tomato TSWV Tomato Particles about Insects (Thrips Mainly in
spotted spotted wilt Æ 7090 nm spp), East of
wilt virus virus with a mechanically Europe and
lipoprotein cuscuta USA
envelope
Tobacco TLCV Geminivirus Germinate Insects Mainly in
leaf curl particles 1520 (Bemisia spp) tropical and
virus × 2530 nm subtropical
countries
Tobacco TVMV Potyvirus Flexuous rod Insects and Serious
vein 765 × 13 nm mechanically disease in
mottle Southeastern
virus USA,
occasionally
in Portugal,
 Columbia,
China
Tobacco TBV Complex Two particles Insects Mainly in
bushy top (1) Æ 213 nm (aphids) Southern
virus (2) Æ 89 nm African
countries
(Zimbabwe,
South
Africa,
Malawi)

phloem and to rapidly reach the growing regions or meristems of plants. The
distribution of viruses within a plant varies with the virus and the plant and is
related to symptom expression.
Transmission and Spread of Viruses
Transmissibility is an essential criterion for all viruses. Nearly all viruses can be
transmitted by grafting, and the majority of viruses can be transmitted by
inoculation of sap. However, relatively few viruses are sufficiently contagious to
be transmitted simply by contact between plant leaves: the exception is TMV
(Zaitlin & Israel, 1975). Such a virus is highly contagious and can be very
difficult to control in susceptible cultivars. Transmission of viruses through the
seed seems to be exceptional, but an increasing number of viruses are now
known to be seed-borne.
Many viruses are transmitted by insects (aphids, thrips, whiteflies, etc.), and
different relationships between the vector and viruses can be defined as follows:

 Page 201
Non persistent is the characteristic of viruses that are acquired by
insects (adult or larvae) and are immediately infectious but infectivity
is soon lost, for example potyvirus (potato virus Y (PVY), tobacco
vein mottle virus (TVMV), tobacco etch virus (TEV), etc.), and
cucumovirus (cucumber mosaic virus (CMV)) (Francki ,et al., 1979;
De Bokx & Hutting, 1981).
Persistent is the term used for viruses acquired only after a prolonged
feeding on an infected plant and a latent period may be necessary
before the insect can transmit.
There is also a category of semi-persistent viruses intermediate
between the two types described above. In addition, the tomato
spotted wilt virus (TSWV) seems to be unique; the adult thrips vector
must acquire the virus in the larval stage in order for the adult thrip to
transmit TSWV (Ie, 1970).
A survey made within the scope of CORESTA (Delon, et al., 1987;
Delon, et al., 1993) identifies 16 viruses responsible for damage on
tobacco, out of which eight (in order of importance) are very
damaging to the tobacco crop: TMV, PVY, tobacco leaf curl virus
(TLCV), CMV, tomato spotted wilt virus (TSWV), TEV, TVMV and
tobacco bushy top virus (TBV). These eight tobacco virus diseases
will be described in more detail below.
Tobacco Mosaic Virus (TMV)
a
Basic Description
Tobacco mosaic virus is a member of the tobamovirus group. The
virus particles or virions are rigid rods of 18 nm in diameter with one
modal length of 300 nm. They are built from a single species of
protein subunit arranged in a helix; each particle contains a single
molecule of positive sense, single stranded RNA (Zaitlin & Israel,
1975). TMV has very stable particles that usually occur in high
concentrations in their host plants. It is easily transmitted
mechanically and naturally by contact and incidental wounding. TMV
is also a good immunogen.
TMV can be transmitted to numerous species in 30 angiosperm
families, but it is only important economically to some members of
the Solanaceae. Outside tobacco, TMV can naturally infect tomato,
pepper and wild Solanaceae (Zaitlin & Israel, 1975).
b
Geographical Distribution
TMV occurs worldwide in all tobacco growing countries (Zaitlin &
Israel, 1975; Delon, et al., 1987; Delon, et al., 1993).
c
Diseases Symptoms
TMV induces classic mosaic symptoms on tobacco, characterized as a
mottled appearance of the leaf with alternating areas of light and dark
green tissues (Fig. 6.2). This symptom is especially noted in the top of
the plant or in younger tissue. The first signs of infection are vein
clearing followed by vein banding and mosaic. In the case of severe
early infections, plants are usually stunted and the lamina of the leaf is
considerably reduced, giving a filiform appearance. During periods of
high temperature and high light intensity, affected portions of leaves
may die. This is called a 'mosaic burn'.
d
Epidemiology
TMV is a very contagious disease that can occur in seedbeds as well
in the field. It is one of the most infectious and persistent disease
agents and is
 Fig. 6.2
Large vein banding on leaves of flue-cured
tobacco (tobacco mosaic virus (TMV) Courtesy
D. Blancard).

 Page 202
mechanically transmitted. It can be readily spread among plants by
workers as they perform operations in the plant beds or fields. TMV
can also be carried in tobacco products such as cigarettes, cigars or
chewing tobacco. It can survive for many years in plant debris which
can then serve as a source of inoculum.
There is little or no evidence that tobacco strains of TMV are seed
transmissible, although tomato and pepper strains of this virus are
frequently transmissible by seed (Lucas, 1975). TMV is not as
sensitive to weather conditions as most other diseases. However, it is
easier for infection to take place where there is moisture on the plants
and when plants are succulent and growing rapidly.
e
Economic Importance
A high disease incidence of TMV infection in tobacco can result in
low yields and poor leaf quality. The greatest crop losses due to TMV
infection have been reported in areas of North, Central and South
America and in Asia (Delon, et al., 1987; Delon, et al., 1993). In
North Carolina, the estimation of annual losses in value caused by
TMV range from 0.03% to 0.88% (Johnson, et al., 1983) and the
monitoring of the loss due to TMV in North Carolina shows an
increasing incidence of this virus in recent years.
In a survey made by CORESTA (Delon, et al., 1993), TMV was found
to be less damaging than in previous years; this is primarily due to the
cultivation of resistant varieties. Nevertheless, we must not forget the
highly infectious nature of this disease and the grave potential for
disease development in susceptible crops.
f
Means of Control
Because of its contagious nature, control of TMV must be approached
very diligently. The most efficient technique of controlling TMV is to
keep the crop virus-free. To prevent the mechanical spread of TMV in
transplants, spray the plant bed with milk (20 L whole milk mixed
with 20 L water applied to 100 m2) or have all workers dip their hands
in milk or a strong solution of a phosphate detergent every 20 to 30
minutes (Lucas, 1975; Shew & Lucas, 1991). This may prevent the
spread of TMV from plant to plant and from seedbeds to the field.
Sanitation practices such as roguing infected plants, destroying stalks
and roots and crop rotation may also contribute to the reduction of
TMV.
The use of resistant tobacco varieties could be recommended as the
best tactic for managing TMV disease. The genetic factor responsible
for the resistance to TMV is derived from N. glutinosa (Wernsman,
1992). It is a single dominant gene (N) which restricts the virus to
localized necrotic lesions (hypersensitive reaction) that cause little
damage to the leaves. Holmes (1936) made the initial transfer of the N
gene from N. glutinosa to N. tabacum. Many dark air-cured and
burley varieties with this resistance gene are cultivated throughout the
world.
For flue-cured varieties, TMV resistance seems to be related to poorer
leaf quality ('off color' and lower grade index). Nevertheless, Johnson
and Main (1983) have shown that recently released TMV flue-cured
varieties demonstrate significantly improved yield and qualities over
their predecessors.
Potato Virus Y (PVY)
a
Basic Description
PVY is the type member of the potyvirus family. Like other
potyviruses it has filamentous, flexible particles 730 × 11 nm in size.
This virus is easily transmitted mechanically to a narrow range of
hosts (De Bokx & Hutting, 1981).
The biological and serological characterizations of the various PVY
isolates have been mostly carried out using the potato. In tobacco,
PVY strains are broadly classified in two types:
A moderate strain that produces mosaic-type symptoms (vein
banding, vein clearing, chlorotic marbling).
A severe strain that causes necrotic areas between the veins as well as
mosaic-type symptoms (Park, et al., 1984). These necrotic symptoms
have been reported in many countries and they are by far the most
destructive (Izard, 1964; Gooding & Lap, 1980) (Fig. 6.3).
There are few precise and detailed classifications of PVY strains apart
from their symptoms on tobacco. Only Gooding and Tolin (1973) in
the USA characterized three isolates according to their reaction on
root knot nematode (Meloidogyne incognita) susceptible (McNair
944) and resistant (NC 95) flue-cured tobacco. These authors
observed in the field that root knot resistant genotypes show serious
necrosis in the presence of a particular isolate of PVY (Gooding,
1985). The three isolates were defined as:

 Page 203

Fig. 6.3
Some necrotic lines along the vein of old leaves
of dark air-cured tobacco (oak leaves) characterize
a CMV contamination (Cucumber Mosaic Virus,
CMV) (Courtesy D. Blancard).
strain MSMR causing the symptoms of mosaic type vein banding,
clearing of the veins on both root-knot susceptible and resistant
genotypes MN 944 and NC 95, respectively;
strain MSNR causing mosaic on root knot susceptible MN 944 and
very necrotic symptoms on root knot resistant NC 95;
strain NSNR which causes necrosis in both root knot susceptible and
resistant genotypes.
It has been reported that the necrotic response of root knot nematode
resistant cultivars to strains MSNR of PVY is due to a pleiotropic
effect of the gene conditioning root knot resistance (Rufty, et al.,
1983).
Other observations on the pathogenicity of PVY isolates have been
reported, such as a necrotic strain capable of overcoming the recessive
gene (va), derived from the genotype known as virgin A mutate
(VAM), for resistance to PVY (Blancard, et al., 1995). This has been
demonstrated in greenhouse evaluations of an American NN strain of
PVY. This isolate has been named VAM-B (virgin A mutant breaking)
strain because it induces symptoms on genotypes that possess the
VAM resistance to PVY. A strain with similar characteristics has
occurred naturally in a field in the USA (Reddick, et al., 1991).
Strains with similar virulence have also been reported in Hungary
(Horvath, 1993), Poland (Glazewska, 1977), Italy (Piccirillo & Piro,
1986) and France (Blancard, et al., 1994). It has also been found in
other countries on the American continent, especially in Chile
(Latorre, et al., 1982) and Argentina (Gooding, 1985).
b
Geographical Distribution
Potato virus Y is found worldwide. Tobacco grown in Europe is
particularly affected by this virus. The very aggressive necrotic strains
have been reported in many East European countries (Hungary,
Germany, Poland, etc.) and also in the western portion of this
continent (Spain, Italy, France, etc.) (Piccirillo & Piro, 1986; De La
Puerta, et al., 1991; Blancard, et al., 1994). Asia is also seriously
affected by necrotic isolates and some have been reported to occur in
China, Japan (Tomaru, 1983), Korea (Park, et al., 1984) and Taiwan.
Also, PVY is found in many African countries, particularly in
Morocco in North Africa (Lockhart & Fisher, 1976). In North
America the situation seems to be completely different; if PVY is
frequently found in crops the most widespread strains seem to be of a
less pathogenic nature than those found in Europe (Henderson &
Troutman, 1963; Lap & Gooding, 1976; Gooding & Lap, 1980; Pirone
& Nesmith, 1994). This does not seem to be the case in South
America and particularly in Argentina and Chile (Latorre, et al., 1982)
where severe epidemics of necrotic strains have occurred.
c
Disease Symptoms
The symptoms caused by PVY on tobacco are much more numerous
and diverse than the American name vein banding would suggest. The
following symptoms can be observed:
Different abnormalities of color in the leaves of affected plants with
more or less pronounced marbling between veins. Veins tend to
yellow while sections of the lamina stay greener. Sometimes many
rings or bright yellow spots can be observed on leaves in the middle
of the plant.
Necrosis can be observed in different forms including browning of the
mid-rib and/or secondary veins of the leaf. This syndrome is the origin
of the European name of this virus 'brown rib', 'Tabak-Rippenbraune',
'maladie des côtes brunes'. When the attacks are very serious, the
lignified vessels, the stalk and the pith, show brown or black necrosis.

 Page 204
White spots, but more often beige or brown ones, of different sizes
often begin near the veins.
For some genotypes the foliar necrosis is more superficial and less
diffused, giving a brassy look to the lamina. This symptom is due to
the death of cells in the mesophyll of virus-infected tobacco leaves
exposed to the sun (Lucas, 1975). Deall (1980) attributes this
symptom to the intervention of a particular strain of PVY expressed in
ripe leaves. The affected leaves are sometimes reduced in size and are
crisp and rolled. If infection occurs early, the growth of plants is
severely reduced. In general, the nature and intensity of these
symptoms vary according to the type of tobacco cultivated, the variety
and, above all, the nature of the virus strain infecting the plant.
Spectacular symptoms, sometimes lethal ones, can result from mixed
infection of PVY with other viruses, particularly CMV.
d
Epidemiology
The host plant spectrum of PVY seems to be limited to cultivated
Solanaceous crops, potato, pepper, tomato and tobacco as well as
different weed species (Solanum nigrum, Portulaca oleracea, Senecio
vulgaris, Physalis spp.). This diversity helps assure, among other
things, the preservation of PVY during the winter months.
At least 25 species of aphids are able to transmit PVY in a non-
persistent manner. Myzus persicae, the green peach aphid, seems to be
the most common and frequent vector in temperate regions. The virus
is rapidly acquired by the insect as it feeds directly in the phloem, and
very few viral particles are needed to assure efficient transmission
(Gooding, 1971, Pirone & Thornbury, 1988). However, for
transmission by aphids the presence of a protein of viral origin is
necessary. This protein has been termed the helper component or the
assisting factor. Only the particles acquired in the presence of this
factor are retained by the stylet of the aphid and are transmissible. The
efficiency of transmission depends on the aphid vector as well as on
the virus isolate.
Mechanical transmission of PVY by inoculation to different members
of the Chenopodiacea, Amaranthacae, Leguminoseae, Compositae
and Solanaceae families (Richter, 1988) is possible. It has not been
reported to be seed transmissible.
e
Economic Importance
At present PVY is certainly the virus which causes the most damage
to tobacco crops worldwide. This fact was confirmed by the latest
CORESTA survey on tobacco viruses. Furthermore, production losses
caused by this virus, especially the necrotic strains, are increasing in
many countries (Delon, et al., 1993). Repercussions of this virus on
burley and Virginia tobaccos are a reduction in the size and weight of
the leaves, plant height and yield. The earlier the infection occurs the
worse is the effect on the crop. In some highly infected fields a
reduction of more than 70% in yield has been noted in Chile (Latorre
& Flores, 1985) and in New Zealand (Thomson & Wright, 1966). The
quality of the tobacco harvested also greatly decreases. An increase in
nicotine, nornicotine, total nitrogen, acid-insoluble nitrogen, and
nitrates can result from PVY infection (Thomson & Wright, 1966;
Sievert, 1978; Latorre & Flores, 1985).
f
Means of Control
The best method to control PVY is to use resistant varieties,
particularly in regions where the virus is present. Resistant varieties
have been grown for many years. Since the first appearance of
necrotic strains of PVY (PVY N) in Europe, it has been noted that
many resistant varieties were traditionally cultivated. Because of the
serious nature of these PVY N strains, and in particular their reaction
on Virginia types, work on improving germplasm has been initiated.
Koelle (1958) obtained a mutant line (VAM), using radiation by X-
rays, that showed good resistance to necrotic PVY isolates. The
resistance of VAM was later reported to be governed by a recessive
gene (Koelle, 1961). Further selections were made to increase this
disease resistance, and these selections such as SCR (Aurea) and
Perevi, Polalta and Havana II C were later used as sources of
resistance in many countries.
Many countries in the world have used resistant European cultivars
(Paraguay, Alta, SCR, Perevi) or VAM as a source of resistance to
PVY disease development in tobacco. All these resistances seem to be
governed by a recessive gene situated in the same locus, but this gene
appears to have different allelic forms as evidenced by the variation in
expression of resistance.
Although the VAM source of resistance still appears to be effective
against the South American strains, the strategy of selection for PVY
resistance should now include the American VAM-B and European
strains. Another source of PVY resistance has been obtained in the
USA by androgenesis in vitro from the PVY sensitive cultivar Mc
Nair 944. This variant has a semi-dominant gene called vr that gives

 Page 205
resistance to necrosis but is only effective against some strains of
PVY especially the VAM-B strains (Witherspoon, et al., 1991).
However, many European strains of PVY induce serious necrosis on
this genotype. Work is currently under way to incorporate the
resistance of Nicotiana africana which is governed by a dominant
gene (Keum, et al., 1991). At present, the introduction of this
resistance into Nicotiana tabacum is difficult due to the low frequency
or lack of pairing between the chromosomes of the two genomes. A
large number of Nicotiana spp. are also resistant to PVY and therefore
could serve as possible sources of resistance.
Molecular biology has opened a fabulous array of prospects for the
fight against this virus (Dougherty & Carrington, 1988) and could
ultimately provide a solution to the management of VAM-B strains
and their relatives.
All measures necessary to stop, or at least limit, the introduction of
PVY in tobacco fields should be taken into consideration. In countries
where infection occurs very early, seedbeds and transplants should be
protected. A physical barrier of nonwoven fabric covering transplants
or greenhouse openings should delay contamination by vectors. Plants
showing symptoms of PVY should be eliminated rapidly, and in no
case should infected plants be replanted later. Careful weeding of the
plot and its surroundings (paths, hedges and roads) should be carried
out to remove sources of the virus and/or vectors. Tobacco should not
be planted near susceptible PVY crops such as potato, tomato and
pepper. Like tobacco, tomato seems capable of harbouring all PVY
strains and acts as a universal host.
Aphid infestation of tobacco should be controlled, but unfortunately,
this has not been shown to be very efficient in combating epidemics of
this virus. In fact, aphid vectors frequently come from outside the
tobacco field and transmit the virus during a very brief injection
period before the insecticide has had time to act.
Tobacco Leaf Curl Virus (TLCV)
a
Basic Description
This virus consists of two incomplete icosahedrons which form a
geminate particle 15 to 20 × 25 to 30 nm in size, and hence belongs to
the gemini group of plant viruses. The genetic material is DNA (Osaki
& Inouye, 1981).
b
Geographical Distribution
The occurrence of TLCV is restricted to the tropics and subtropics,
although it has also been reported in some temperate areas, e.g. Japan,
parts of Europe and the USA (Osaki & Inouye, 1981).
c
Diseases Symptoms
A number of different TLCV strains have been reported to cause a
wide variety of disease symptoms on various hosts (Osaki & Inouye,
1981). Symptoms on tobacco include: stunted plants with twisted
stems and leaves (Fig. 6.4); dark green thickenings or enations along
veins on the underside of the leaves (Fig. 6.5); small puckered leaves
with downward-rolled leaf margins; and plants with knotted or
crooked midribs and veins (Lucas, 1975; Osaki & Inouye, 1981). For
a milder form of the virus, these symptoms may only be visible in the
upper parts of the plant (Lucas, 1975). Symptoms generally appear 3
to 4 weeks after infection has taken place (Lucas, 1975).
 Fig. 6.4
Necrotic strains of PVY cause browning of midribs
and veins and necrotic rings (Potato Virus Y, PVY)
(Courtesy D. Blancard).
d
Epidemiology
This virus is transmitted primarily by the whitefly Bemisia tabaci.
Seed and sap transmission have not been reported, but the virus is
graft transmissible. As

 Page 206

Fig. 6.5
Tobacco leaf curl virus causing twisted stems
and leaves.
this virus has never been reported to be sap transmissible it cannot be
mechanically inoculated (Lucas, 1975; Osaki & Inouye, 1981).
Bemisia tabaci requires an acquisition time of 15 minutes to 2 hours
on infected hosts, and a transmission time of 10 to 60 minutes on
healthy plants (Lucas, 1975; Osaki & Inouye, 1981). In dry seasons
when whiteflies are prevalent, the disease incidence of TLCV is
increased, as is the case when any of the 63 species of overwintering
hosts are present (Smith, 1957; Lucas, 1975). The disease has rarely
been observed in seedbeds (Lucas, 1975).
e
Economic Importance
Leaves of infected plants are generally small, deformed and not
harvestable (Lucas, 1975). Heavy losses occur in South America,
Asia, Australia and Africa (Delon, et al., 1993). Plants infected with
TLCV are also more susceptible to infection by other pathogens,
particularly Cercospora (Lucas, 1975).
f
Means of Control
Whitefly control with insecticides and the removal of overwintering
hosts are the only effective means of controlling the incidence of
TLCV. Growers must ensure that healthy seedlings are planted and
that infected plants are destroyed (Lucas, 1975).
Cucumber Mosaic Virus (CMV)
a
Basic Description
CMV is a member of the cucumovirus group. It has a hollow
icosahedric particle 28 to 30 nm in diameter with a capsid constituted
of an assembly of 180 subunit proteins. Its genome is composed of
three main molecules of RNA, monocatenair positive designated as
RNA 1, RNA 2 and RNA 3 according to their decreasing molecular
weight. RNA 1 and RNA 2 are separately encapsided; RNA 3 is
encapsided with a four sub-genome of the same sequence as the
extremity 3 of RNA 3 which is not necessary for infection. The RNA
monocistronic 1 and 2 coding for proteins P1 and P2 are involved in
the replication of the virus (Nitta, et al., 1988).
The induction of symptoms is the result of the effect of the RNA
satellite on the host plant with the interaction of the virus helper and
not the direct effect of a protein coded by the RNA satellite. It is more
susceptible to modification by environmental conditions, especially
temperature (Kaper, 1992).
CMV isolates can be differentiated according to their function and
their biological characteristics (symptomology and thermosensibility)
by serological criteria and molecular properties (Table 6.2).
Table 6.2 Main names given to the two principal
groups of CMV strains.
Group Criteria of Authors
characterization
C B Symptoms Marrou, et al.,
1974
thermo- thermo- Thermal Marchoux, et
resistant sensitive al., 1976
U N Serological Richter, et al.,
1972
DTL ToRS Serological Devergne &
Cardin, 1975
WT S Serological Piazzolla, et al.,
1979
I II Serological Owen &
Palukaitis, 1988

b
Geographical Distribution
CMV has a worldwide distribution (Francki, et al., 1979), but its
incidence on tobacco varies widely from one country to another. It is a
particularly serious disease of tobacco in Asia and China (Han, et al.,
1983). It has been reported on tobacco in Europe, Japan and

 Page 207
Taiwan (Marte, 1980; Piccirillo & Tonini, 1990; De Los Angeles, et
al., 1992; Blancard, et al., 1994) alone or in a complex with other
viruses. CMV is present in other countries such as the USA (Gooding,
1971), India and The Philippines, but disease caused by CMV is
minor in these countries and appears to have little effect on tobacco
production.
c
Disease Symptoms
The symptoms caused by CMV differ significantly according to the
type of tobacco infected. They can be confused with symptoms
provoked by other viruses such as TEV and TMV (Pena-Inglesias, et
al., 1982), AMV (Lucas, 1975) and TRSV. Indeed, quite different
symptoms in infected plants can be fairly systematically observed on
all types of tobacco. The symptoms include the following:
A more or less severe mosaic, sometimes accompanied by vein
banding and or inter-rib yellowing. Strains responsible for very yellow
to white mosaic of the 'Aucuba' type have been reported on many
occasions in the literature (Marchoux, et al., 1976; Wan, et al., 1984).
Different anomalies of the lamina such as the presence of blisters, a
tendency for leaves to be filiform or shriveled.
Localized necrotic alterations taking the form of small beige or brown
lines (etches) arranged in lines or rings. Chlorotic and necrotic lines
sometimes trace the ribs of the lamina giving an oak leaf symptom.
This symptom is found on the bottom leaves of dark air-cured tobacco
(Fig. 6.6).
 Fig. 6.6
Leaf enations (outgrowths) caused by tobacco
leaf curl virus.
General necrotic alterations bringing about the destruction of the
terminal bud. These alterations are accompanied by necrosis present
on the leaves, ribs, stalk and pith. These symptoms, which sometimes
affect a large proportion of the plants, are mainly found in dark air-
cured tobacco. They can be confused with those caused by TSWV.
Verhoyen (1962) had already described these types of symptoms
which he sometimes attributed to the virus complexes: TMV + PVY-N
or TMV + CMV. The necrotic strains of CMV on tobacco have also
been described in the Philippines (Revellame, et al., 1982) and in
France (Blancard, et al., 1994).
Mixed infections are quite frequent in many countries, associating
especially CMV and PVY. Symptoms from this combination are even
more spectacular (Marte, 1980), sometimes resulting in lethal necrosis
(Han, et al., 1983).
d
Epidemiology
CMV is capable of infecting a large number of hosts, including 191
species belonging to 40 families (Francki, et al., 1979). This large
spectrum partly explains why CMV has reappeared from year to year,
as it is probably maintained in weed hosts. Cultivated plants such as
spinach, winter celery, different curcubits (melon, cucumber,
courgette) and other Solanaceae (tomato, pepper) also serve as
reserves for CMV.
CMV is transmitted from infected plants to healthy plants by aphids in
a nonpersistent fashion. The aphids rapidly acquire virus particles in
their stylets and teguments of the mouth. Virus acquisition can occur
during a short period of not more than 2 to 4 hours. About 60 species
of aphids are able to transmit CMV (Francki, et al., 1979). Different
species of aphids vary in their ability to transmit CMV. The best
vectors are Myzus persicae, Aphis gossypii, A. craccivora and A.
fabae (Mansion & Dunez, 1988).
Seed transmission has been reported for some weeds and 20 vegetable
species (Kaper & Waterworth, 1981). In Stellaria media, CMV is
present in the seeds, therefore transmitting the disease to its progeny.
This must influence the epidemiology of this virus. The spread of
CMV depends on the epizootic nature of the aphids. Indeed, many
abiotic factors play an essential role in the biological efficiency of
these insects. For instance:
wind determines direction of aphid flight;
temperature influences growth of the crop and the progression of
disease development within a plant;

 Page 208
tobacco production practices;
the proximity of other virus-infected source plants.
e
Means of Control
In countries where infection occurs early in seedbeds, young plants
should be protected. This can be done with nonwoven tissues, (Agryl
P17TM, ReemayTM, etc.), which act as physical barriers to the insect
vector. Plants that have symptoms of CMV should be rapidly
eliminated and at no time should infected plants be transplanted into
the field. Careful weeding of the plot and its borders (hedges and
tracks) should be carried out to eliminate sources of viruses and/or
insect vectors. Planting tobacco near CMV susceptible plants such as
tomato, spinach or cucurbits should be avoided.
Special precaution must be taken to control aphid populations on
tobacco. Unfortunately, these methods are often ineffective in
controlling virus epidemics. In fact, aphid vectors frequently come
from outside the plot and transmit the virus by a brief injection before
the vector is killed by the insecticide.
Resistance to CMV infection in tobacco has been discussed (Fulton,
1953; Troutman & Fulton, 1958). In many countries where CMV is
severely endemic, there are no CMV resistant tobacco varieties. A
program of selection for CMV resistance has been initiated in Taiwan
(Wan, et al., 1984), using a source of resistance developed in the
USA, TI 245 (Holmes, 1960; Holmes, 1961; Gooding, 1971). The
partial resistance given by seven genes has given a reduced rate of
disease development in the plants. A more efficient source of
resistance than this has been described in an Indian tobacco, but has
never been used (Thomson & Wright, 1966). Many other genotypes of
tobacco in Japan with more varying resistance to CMV (Oka &
Yoshino, 1988) have been described.
f
Economic Importance
It is never easy to determine with certainty the incidence of a disease
on a crop and, more particularly, the extent of damage it may cause.
However, research in Italy on field crops has shown a high incidence
of CMV on tobacco (Piro, et al., 1990). Yield losses in 1989 and 1990
were 46% and 30%, respectively, which was greater than losses
attributed to PVY infection (Piro, et al., 1990). Production values
were reduced 50% by CMV and 55% in cases of mixed CMV-PVY
infections. Significant reductions in total alkaloid content were also
noted in tobaccos infected with CMV and CMV + PVY. This disease
is important for tobacco in those countries where it is commonly
found in other crops.
Tomato Spotted Wilt Virus (TSWV)
a
Basic Description
TSWV is a RNA-containing virus with membranebound particles
about 70 to 90 nm in diameter. TSWV is the sole member of this
group. The name Lycopersicon virus 3 is also widely used. TSWV is
physically and chemically one of the most unstable plant viruses, but
is readily sap transmissible when neutral buffers containing reducing
agents are used (Ie, 1970). It has a very wide host range infecting 160
dicotyledons and 10 monocotyledons, especially members of the
Solanaceae, Compositae and Leguminosae (Ie, 1970; Lucas, 1975).
b
Geographical Distribution
First described in Australia in 1919 on tomato, Brillelebank named the
disease 'spotted wilt' (Ie, 1970; Lucas, 1975). TSWV is common in
temperate and subtropical regions throughout the world. It became a
disease of great economic importance to European tobacco producers
around 1941 (Ivancheva-Gabroska, 1979; Mickovski, 1981). TSWV is
also widespread in south-eastern Brazil. In the US, it was identified on
Perique tobacco in Louisiana in 1975 and in Georgia in 1986 where
losses have become more and more significant (Culbreath, 1991;
Culbreath, et al., 1993).
c
Disease Symptoms
Symptoms of TSWV infection are quite variable. Concentric necrotic
rings and zonate necrotic spots appear on the younger leaves (Fig.
6.7). At first, the spots are yellow and they rapidly turn red brown.
Young seedlings to mature tobacco plants may be attacked. Infested
plants are stunted, and the apical bud droops or bends over. Young
leaves infected on one side are frequently distorted or puckered as a
result of unequal growth throughout a leaf. Necrotic streaks develop
along the stem and dark necrotic areas or cavities appear in the cortex
and the pith. Diseased plants rarely yield any curable leaves.

 Page 209

Fig. 6.7
Necrotic and concentric spots localized near the
veins of a leaf of dark air-cured tobacco, these are
also brown (Tomato Spotted Wilt Virus, TSWV)
(Courtesy D. Blancard).
d
Epidemiology
Many minor variants giving symptoms differing in disease severity
have been isolated and TSMV occurs naturally in at least six unique
strains (Ie, 1970; Lucas, 1975; IvanchevaGabroska, 1979).
TSWV is transmitted by thrips: Thrips tabaci, Frankliniella schultzei,
F. occidentalis and F. fusca (Ie. 1970; Lucas, 1975;
IvanchevaGabroska, 1979). The virus is acquired only by the larvae,
but only adults can transmit TSWV. Vectors retain the infectious virus
for all of their life, about 5 to 9 weeks, but do not transmit the virus to
their progeny (Lucas, 1975; IvanchevaGabroska, 1979). There are
many generations (9 to 20) of thrips that can be produced each year.
TSWV has not been found to be seed transmitted (Ie, 1970). Often
tobacco becomes infested with thrips when seedlings first emerge.
Both TSWV and thrips are transplanted to the field with the seedlings.
Warm weather and dry air provide favorable conditions for thrips
multiplication. Disease outbreaks occur in the field during the
vegetative period when the virus is transmitted from plant to plant by
vectors.
Overwintering of the virus may occur in several wild plant species.
However, the level of virus transmission depends greatly on the vector
populations. The main vector Thrips tabaci overwinters as an adult
insect in the soil; in spring when soil temperature increases, the vector
moves to weeds.
e
Economic Importance
The wide distribution of TSWV becomes less important when
considering the large yield losses it causes in East European countries
(Delon, et al., 1993). In Bulgaria, damage was particularly severe in
1956 and 1969 where losses were estimated to be worth more than
$20 million USD (Ivancheva-Gabroksa, 1979). In 1969, Greece
(Katis, et al., 1992) and Yugoslavia (Mickovski, 1981) reported
epidemics of the disease. All tobacco types are susceptible to TSWV,
and leaves of infected plants are often of poor quality.
f
Means of Control
The most effective technique of controlling TSWV has been by
managing the insect vectors. Chemical control, especially using
recommended insecticides during seedling growth, is one important
method of TSWV disease control (Tsakiridis, 1983; Mazur, 1985).
Tactics such as rotating with nonsusceptible crops or destroying
diseased plants are useful but not always sufficient. TSWV resistant
tobacco cultivars are being developed by incorporating the disease
resistance gene found in N. alata into cultivated tobacco (Gajos, 1987;
Gajos, 1988; Nielsen, 1993).
Tobacco Etch Virus (TEV)
a
Basic Description
The virus particles of TEV are rod-shaped and flexuous, with a
diameter of 12 to 13 nm and a length of 730 nm (Shepherd &
Purcifull, 1971). TEV is a member of the potyvirus group because of
its similar appearance and other biological and physical properties to
other potyviruses (Lucas, 1975; Shepherd and Purcifull, 1971).
Pinwheel (cylindrical) cytoplasmic inclusions and other crystalline
nuclear inclusions are present in the cells infected with some strains of
TEV, and can be seen under a light microscope (Smith, 1957;
Shepherd & Purcifull, 1971; Lucas, 1975). The par-

 Page 210
ticles contain single-stranded RNA as the genetic material (Shepherd
& Purcifull, 1971). The virus is easily transmitted mechanically to
other plant hosts (Shepherd & Purcifull, 1971). TEV has several
solanaceous overwintering hosts, which include tomato, petunia,
potato and Datura stramonium (Smith, 1957; Shepherd & Purcifull,
1971).
b
Geographical Distribution
TEV is generally confined to North and South America, particularly
in parts of Canada, USA, Mexico, Puerto Rico and Venezuela
(Shepherd & Purcifull, 1971). However, serious losses due to the virus
have been reported in Madagascar, occasionally in South Africa and
in parts of the Far East (Delon, et al., 1993).
c
Disease Symptoms
Not unlike many plant viruses, symptoms vary in severity depending
on the strain of the virus (Lucas, 1975). Disease symptoms generally
become apparent near flowering in the case of natural infections and
within 6 to 10 days after inoculation in the greenhouse (Smith, 1957;
Lucas, 1975). Symptoms include faint, interveinal chlorotic spots
which develop necrotic ring patterns (Smith, 1957); vein clearing and
line patterns of necrosis; in the lower leaves and roots necrosis occurs
3 to 4 weeks after infection (Lucas, 1975). These symptoms generally
begin at the base of the leaves and spread to the leaf tips (Smith,
1957). Plants that are severely infected with TEV are often stunted
due to shorter internodes between the leaves (Smith, 1957; Lucas,
1975). Leaf 'firing', chlorosis, and tattering at final harvest cause a
reduction of leaf quality (Lucas, 1975). Strains of the virus are
differentiated on the basis of the type and severity of the expressed
symptoms:
K severe strain;
N strain causes strong necrosis, no cellular inclusions;
V strain causes vein clearing and mottling, no inclusion present
(Smith, 1957).
d
Epidemiology
TEV is transmitted nonpersistently by aphids, particularly Myzus
persicae (Shepherd & Purcifull, 1971), with the transmission being
primarily by mechanical effects of the mouthparts (Lucas, 1975). This
virus has not been found to be seed transmissible (Smith, 1957).
e
Economic Importance
The amount of damage caused by the virus fluctuates yearly due to the
shifts in the aphid populations and the amount of overwintering
inoculum. TEV causes significant yield losses throughout the burley
and fluecured tobacco producing regions in the southeastern USA
(Gooding & Ross, 1970).
f
Means of Control
Like other aphid-transmitted viruses, it is often difficult to prevent the
introduction of the virus into the crop. The virus often overwinters in
several weed hosts and is then transmitted by aphids. The most
effective means of controlling TEV is by controlling or monitoring the
aphid populations and by growing disease resistant varieties. In 1986,
a burley variety was released in the USA having medium-high
resistance to some isolates of TEV. The source of resistance is from
the tobacco introduction (TI) 1406 or VAM (Miller, 1987; Reddick, et
al., 1991). As reported earlier, this line was first introduced as a
source of PVY resistance.
Tobacco Vein Mottling Virus (TVMV)
a
Basic Description
This virus is a member of the potyvirus group, having a flexuous
filament 765 nm in length with a diameter of 13 nm. It is a single-
stranded RNA virus (Pirone & Shaw, 1988).
b
Geographical Distribution
Worldwide there are relatively few incidences of TVMV, but it does
occur occasionally in Portugal, Columbia and China (Delon, et al.,
1993). TVMV is considered to be a serious disease of burley tobacco
in the southeastern USA (Pirone & Shaw, 1988).
c
Disease Symptoms
TVMV was first described in 1972 on burley tobacco in North
Carolina (Gooding & Sun, 1972). Diseased tobacco had symptoms
similar in character with viruses (PVY, TEV) previously described in
the potyvirus group. The virus was named after its striking
veinmottling type symptoms (Sun, et al., 1974). Initial symptoms
consist of faint vein clearing, a discontinuous pattern along the veins
developing into a

 Page 211
mottling or banding (Lucas, 1975; Pirone & Shaw, 1988). Systemic
veinal and laminar necrosis may occur in older plants (Lucas, 1975;
Pirone & Shaw, 1988). In some tomato cultivars, the virus can cause
symptomless infections (Pirone & Shaw, 1988).
d
Epidemiology
The virus can be transmitted mechanically, but is generally
transmitted nonpersistently by aphids. Several aphid species are very
efficient transmitters, including Myzus persicae (Lucas, 1975; Pirone
& Shaw, 1988). Acquisition times of 15 to 30 seconds are followed by
retention times of up to several hours (Pirone & Shaw, 1988). The
virus has a very limited host range (Pirone & Shaw, 1988) and can
only overwinter in perennial solanaceous weeds (Lucas, 1975). Seed
transmission has not been reported (Pirone & Shaw, 1988).
e
Economic Importance
Yield losses vary according to the severity of the isolate, with
different strains producing slightly differing symptoms. Large losses
can result from veinal and laminar necrosis of the leaves (Pirone &
Shaw, 1988). In the late 1970s, TVMV disease incidence was
estimated to be as high as 12% of the North Carolina burley crop
(Gooding & Main, 1981). However, with the commercialization of
TVMV resistant burley varieties, this virus has become less of a
concern for some growers. TVMV is often found in combination with
TEV on burley tobacco in the USA and is often referred to as the virus
complex. Yield losses due to this complex of viruses have been
estimated to be greater than 60% in susceptible cultivars (Fisher &
Rufty, 1993). TVMV infections appear to be on the increase and are a
growing concern for tobacco producers in Columbia (Delon, et al.,
1993).
f
Means of Control
Control is as for the other potyviruses, i.e. reduction in the numbers of
alternative hosts, destruction of the aphid vectors by the use of
insecticides and tobacco-free periods, and the use of resistant cultivars
(Lucas, 1975).
Tobacco Bushy Top Virus (TBTV)
a
Basic Description
Bushy-top is caused by a mixture of two different viruses which are
related to the tobacco rosette complex (Legge, 1960). The rosette
complex has a veindistorting and a mottle component (Legge, 1960),
and the relationship between the two complexes can be seen in (Fig.
6.8) (Deall, 1977). It was suggested that bushy top complex has the
rosette veindistorting component and a mottle component, which is
either an altered rosette mottle virus or a different strain of it (Legge,
1960; Cole, 1962; Deall, 1977). Two unique virus particles have been
found in TBTV-infected plant material: a veindistorting particle with a
diameter of 2 to 13 nm (Cole, 1962; Deall, 1977) and a second
particle, probably the mottle component, with a diameter of 8 to 9 nm
(Deall, 1977).
 Fig. 6.8
The bushy-top/rosette complex, showing the relationship between the viruses
and their methods of transmission.
b
Geographical Distribution
TBTV was first reported in Zimbabwe in 1958 (Gates, 1962; Legge,
1960; Lucas, 1975) and now occurs throughout southern Africa,
including South Africa, Zimbabwe and Malawi, and has been reported
occasionally in Pakistan and Thailand (Delon, et al., 1993).
c
Disease Symptoms
The leaves of both naturally infected and inoculated plants show
symptoms of vein clearing, mottling, vein distorting and rounding.
The leaves are generally crisp and the shoots very brittle (Lucas,
1975). Axillary bud growth is stimulated, giving the 'bushy'
appearance (Fig. 6.9) which is characteristic of this disease (Gates,
1962; Lucas, 1975; Deall, 1977). Infected plants are stunted, although
the number of leaves remains unchanged (Legge, 1960). Flowering
and seed production is affected, with little or no seed being set. Seed
harvested from TBTV-infected plants are often less viable
(Swanepoel, 1993).

 Page 212

Fig. 6.9
'Bushy' tobacco plant (foreground) and normal
plants (background).
d
Epidemiology
This disease is dependent on aphids, especially Myzus persicae, for
transmission. It cannot be mechanically transmitted (Lucas, 1975).
The vector requires an acquisition feeding time of at least 15 minutes
and will generally only transmit the complex after 24 hours (Legge,
1960). Longer acquisition times increase the infectivity retention time
to several days (Gates, 1962).
Throughout the tobacco growing season the number of aphid vectors
increases and, therefore, the potential for transmission of TBTV also
increases (Legge, 1960). It has been found that the disease incidence
is higher in late planted crops (late November and December in the
southern hemisphere) than in those planted earlier (October and early
November) (Legge, 1960).
All other alternative hosts of TBTV have been found to be
solanaceous (Gates, 1962).
e
Economic Importance
The severity of TBTV on tobacco varies according to the growth stage
of the plant when infection occurs. Symptoms are expressed greatly
when infection occurs in the first 3 to 5 weeks after transplanting.
Symptoms are less pronounced if the infection occurs when plants are
growing slowly and are almost negligible in mature plants. TBTV
causes an overall decrease in the yield as the plant's leaves are smaller
and leaf quality is reduced (Legge, 1960).
f
Means of Control
There are several ways to prevent the spread of TBTV:
plant early this allows the crop to become established when aphid
populations are low (Cole, 1966; Lucas, 1975);
apply chemicals use foliar and soil insecticides to reduce the number
of aphid vectors (Cole, 1966; Lucas, 1975);
rogue unwanted weed hosts and infected tobacco plants this reduces
the potential sources of virus inoculum (Lucas, 1975).
Conclusions
Significant economic crop losses result each year from virus diseases
that infect tobacco in all areas of the world where the crop is
produced.
The principal means of disease control continues to focus on
protective measures, such as crop rotation, destroying alternative
weed hosts, and by controlling the insect vectors. No chemical
substances (viricides) are commercially available and effective in
controlling plant virus diseases. When available, the best strategy for
maintaining virus-free crops is to grow diseaseresistant cultivars.
Because pathogen populations are continuously changing by
producing new strains or variants, the need for new sources of virus
resistance is continually increasing. In the near future, plants that have
been genetically engineered for virus resistance will likely provide
additional sources of disease resistance.
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 Page 216

6C
Nematode Pests of Tobacco
J.A. Shepherd
Tobacco Research Board
Harare, Zimbabwe
Introduction
Plant-parasitic nematodes are found wherever tobacco is grown, but
the severity of the problems they cause will depend largely on climate
and soil type.
Many tobacco-producing countries are close to or within the
intertropical area of the world where the dominant plant parasitic
nematodes are from the genus Meloidogyne, of which the most
important are M. arenaria, M. incognita and M. javanica.
Meloidogyne hapla and some other Meloidogyne species, plus some
species of Pratylenchus, Tylenchorhynchus, Globodera,
Rotylenchulus, Ditylenchus and Aphelenchoides may cause yield
losses in certain restricted areas. Some species of Xiphinema,
Longidorus, Trichodorus and Paratrichodorus are reported to transmit
virus diseases to tobacco.
Meloidogyne
a
Life Cycle
Meloidogyne species are sedentary endoparasitic nematodes. The eggs
are laid by the female in a gelatinous matrix which may be completely
or partially embedded in the root of the host plant. The first moult
occurs while the developing nematode is still in the egg, and the
second-stage juvenile hatches soon after. The second-stage juveniles
are mobile and migrate to just behind the root tip of a host plant where
they penetrate. They come to rest with their heads in the developing
stele near the region of cell elongation and with their bodies in the
cortex, lying parallel with the stele. Here the young nematodes start to
feed, causing the cells on which they are feeding to change into 'giant
cells' which are essential for the successful completion of the
nematode's life cycle. Also, this causes the hypertrophy and
hyperplasia of surrounding cells that create the characteristic
enlargement of the root to form galls. Within the gall the juvenile
undergoes two more moults before becoming the swollen female or
the worm-shaped but usually functionless male. In most of the
important root knot nematodes, so called because of the root galls
formed by the feeding nematode, the females produce the next
generation parthenogenetically, that is, without the sexual
involvement of the male. The life cycle takes about 35 days under
normal conditions.
b
Geographical Distribution
Meloidogyne incognita and M. javanica are the most widely
distributed of the root knot nematode species (Table 6.3). They are
mostly confined to the intertropical zones, (35°N35°S), with the limits
for M. incognita being a bit further north or south than M. javanica.
M. arenaria occurs in much the same areas as M. incognita. Their
relative importance and distribution are largely dependent on the
climate, as M. javanica has a greater tolerance to drought and high
temperature than M. incognita, and is therefore more likely to be
found where the rainfall is seasonal (Daulton & Nusbaum, 1961;
Daulton & Nusbaum, 1962). Meloidogyne arenaria and M. hapla are
the next most widely distributed species, with M. hapla being
confined to the cooler parts of the world where the temperatures may
vary between 0°C and 15°C, such as in the north of the USA, southern
Canada, northern Europe and in Africa above 1500 m.
Changes in cultural practices, such as extending the rainy season by
using irrigation, the crops grown in a tobacco rotation or the
introduction of resistant cultivars, can affect the species distribution.
Meloidogyne incognita is widespread in the USA tobacco-growing
areas, but the spread of M. javanica in Florida and M. arenaria in
North and South Carolina is being encouraged by the widespread use
of the M. incognita resistant tobacco cultivars (Fortnum, et al., 1984;
Rich & Garcia, 1985; Schmitt & Barker, 1988). Meloidogyne
incognita tends to predominate when cotton or maize (corn) are grown
in a tobacco rotation, while M. arenaria is encouraged when soybeans
or groundnuts (peanuts) are grown (Fortnum & Currin, 1993).

 Page 217
Table 6.3 Relative importance of Meloidogyne
species in some tobacco growing countries.
Country Meloidogyne
arenariahaplaincognitajavanica
Africa
Malagasy 1 2
Malawi 3
Nigeria 2
South Africa 1 1 3 3
Zimbabwe 1 3
America
Argentina 1 1
Brazil 1 3 3
Canada 1
Chile 2 2
Colombia 1 1
Cuba 3
Guatemala 3
Mexico 2 3 3
Paraguay 3 3
U.S.A. 2 1 3 3
Asia and Oceania
Australia 1 2 2
Bangladesh 2 1
China 2 1
India 3 3
Japan 2 2
Korea 1
Malaysia 2 2
Pakistan 3 3
Philippines 1 3 2
Thailand 1 2 3
Vietnam 3 3
Europe
Albania 2
Bulgaria 2 2
France 1 2
Germany 2 2
Greece 2 2
Hungary 1 1 2 2
Italy 3 3
Spain 3 3
Yugoslavia 1 2 2
(former)
Mediterranean
countries
Iraq 1 2 3
Morocco 2 2
Syria 3 3
Turkey 3 3
Key: 1 Minor importance. 2 Moderately important
or locally important. 3 Very important.
Adapted from: Survey of Pests and Diseases of
Tobacco and Chemicals Used. CORESTA:
Agronomy and Phytopathology Groups, 1987.

Meloidogyne javanica and even M. hapla are found in North Carolina.

In South Africa in 1961, 73% of females identified were M. javanica
and 4% were M. incognita (Milne, 1961), while in a later survey M.
javanica was found in 62% of populations and M. incognita in 72%,
of which 40% were mixed populations (van Niekerk, 1985). This
change may well have been influenced by the wider use of irrigation.
In Zimbabwe, where rains are normally expected only from
November to March, 99% of infestations were found to be M.
javanica (Martin, 1962). Subsequent work has confirmed this and in
the few cases where M. incognita has been found, the soil has been
kept moist for longer periods by irrigation. In Canada, root-knot
nematodes are rare with M. hapla occurring more frequently than any
of the others.
c
Disease Symptoms
The characteristic symptoms of root knot nematode parasitism are the
galls formed by the roots as a reaction to the invasion and feeding by
the nematode. These can range from small individual galls to severe
coalescing growths associated with restriction of root development.
The galls produced by M. hapla are usually small and often only
affect a limited part of the root system, while M. arenaria galls may
be bead-like and affect a large part of the root system. Meloidogyne
incognita and M. javanica produce large galls which may affect 900%
or more of the root, with M. javanica usually producing the most
galling. Root decay, caused by secondary invasion of the galled tissue
by soil-borne pathogens, often develops in roots severely galled by M.
javanica, M. incognita or M. arenaria, but is not so common in roots
galled by M. hapla.
Poor top growth and wilting on hot days are common symptoms of a
severe infestation. There may also be scorching of the leaf tips and
margins with signs of apparent nitrogen and potassium deficiency.
Characteristically, these symptoms are patchily distributed throughout
the field, as a uniform severe infestation is rare. Weeds, which are
usually suppressed by wellgrown tobacco plants, are able to grow and
compete successfully for soil moisture and nutrients.
The galls, in particular the giant cells, provide a favored site for the
development of soil-borne fungi, often enabling the pathogen to
overcome resistance that the plant should have in the absence of
successful root knot nematode invasion. Black shank (Phytophthora
nicotianae), fusarium wilt (F. oxysporum var. nicotianae) and black
root rot (Thielaviopsis basicola) are important diseases that are found
in many tobacco growing areas


 Page 218
of the world, and whose severity is enhanced by root knot nematode
attack. The control of these diseases relies heavily on successful
nematode control (Sasser, et al., 1955; Porter & Powell, 1967; Lucas,
1975; Tu, et al., 1995). Plants heavily infested with root knot
nematodes are more likely to be severely affected by the 'brown spot'
disease caused by Alternaria alternata (Powell & Batten, 1969), and
early infestation may increase the incidence of stem damage by the
sore shin fungus, Rhizoctonia solani (Hartill, 1968; Batten & Powell,
1971). Many other soil fungi that do not normally cause disease on
their own can invade galled roots and enhance the damage by
increasing the root necrosis (Powell, et al., 1971).
d
Pathotypes
All four pathotypes of M. incognita have been recorded on tobacco,
but by far the commonest is race 1 (Taylor, et al., 1982). Race 1 is the
commonest in North Carolina, although races 2 and 4, which can
cause galls on the M. incognita resistant cultivars, and race 3 are also
found. All four races have been reported from Brazil and in India
(Sharma & Gill, 1992). In Zimbabwe races 1 and 3 are found, of
which the commoner is race 3; while in South Africa races 2 and 4
have been identified, of which race 4 is the more commonly found
(van Wyk, 1985). Race 2 of M. arenaria is the predominant pathotype,
when this species is found on tobacco (Taylor, et al., 1982). So far it is
believed that there is only one race of M. javanica and M. hapla.
e
Survival and Dissemination
All the root knot nematodes that attack tobacco have a wide host
range and can survive between tobacco crops on many weeds and
other crops, especially if tobacco is frequently grown in the same
field. The gelatinous material of the egg mass and the undecayed roots
of the host plant provide a protection from desiccation which can
protect the eggs in the event of drought. In Ghana it was found that
many eggs would still hatch even if kept in soil at 3.7% moisture for 5
days, while if undecayed roots were present, the eggs remained viable
at even lower moisture levels (Peacock, 1957). Meloidogyne javanica
egg masses from Zimbabwe, where the soil is usually wet for 4 to 5
months during summer and dry for a similar period during winter,
were able to withstand soil moistures of 3.4% and 20.4% for longer
than M. javanica egg masses from Georgia, USA, where the rainfall is
more uniform (Daulton & Nusbaum, 1962). Root knot nematodes can
also be spread by irrigating with river or dam water that is
contaminated with eggs and juveniles washed into the water from
infested fields.
f
Economic Importance
The three most commonly found root knot nematodes, M. arenaria,
M. incognita and M. javanica, are always pests of economic
importance in tobacco culture, wherever the climate and soil type
favor their development (Barker, et al., 1981). They are of less
importance in cooler areas, such as Canada or France, and tobacco
grown in heavy clay soils often suffers less yield loss than when
grown in a sandy soil (Figures 6.10 and 6.11), (Barker, et al., 1981).
The relative aggressiveness of the commonly found species is shown
by the percentage loss in yield for each 10-fold increase in the initial
population; M. hapla is the least aggressive with only 3%, while
losses with M. incognita are 8 to 9%, M. arenaria 16 to 17% and M.
javanica 19%. Although of less importance, similar losses occur with
the root knot resistant cultivars, such as Speight G28, where a 3% loss
for each 10-fold initial increase of M. incognita race
 Fig. 6.10
Regression of root-galling vs. tobacco yields in
sandy soil in North Carolina, USA.

 Page 219

Fig. 6.11
Regression of root-galling vs. tobacco yields in
a heavy fine textured soil in North Carolina, USA.
3 has been reported (Barker, et al., 1981). This damage response may
well be caused by the hypersensitive reaction of the resistant cultivars
to the nematode invasion, especially at high inoculum levels (Sosa-
Moss, et al., 1983).
The main effect of root knot nematode attack is loss of yield, but this
is usually accompanied by a loss in quality, particularly if the
infestation is severe. Leaves from damaged plants are often smaller
and may have necrotic damage to the tips and edges, and damage
from leaf disease such as Alternaria is often increased.
Estimates of losses vary considerably. An estimate of financial loss
due to root knot nematodes in North Carolina was about US$4000000,
which was about 0.4% of the total value of the crop, despite the
widespread use of M. incognita resistant cultivars and nematicides
(Melton, et al., 1995). In the Mediterranean area M. incognita can
cause losses of 30 to 75%, especially in Oriental cultivars (Lamberti,
1979). In India, losses of between 15 and 50% of cured leaf yield have
been reported, depending on the severity of the infestafion (Hussaini,
1986). In Zimbabwe, in 1963, when over 52% of the crop was
fumigated, it was estimated that losses were between 8 and 11 million
kilograms annually and that field fumigation could cause increases of
between 55 and 1800 kg/ha of cured leaf (Daulton, 1963). Subsequent
experiments have confirmed these estimates and shown that under
severe infestation conditions even greater yield decreases can be
expected.
The threshold damage levels for these nematodes are low. The
Nematode Advisory Service of the North Carolina Department of
Agriculture recommends that if as few as 10 to 200 M. incognita
juveniles per pint of soil are found in a soil sample taken in the fall, a
resistant cultivar should be used, while if more than 200 are found, a
reliable nematicide should be used as well. If root knot nematodes that
break the resistance of the available cultivars are found, the
nematicides become even more important (Melton, et al., 1995). In
Zimbabwe, no threshold levels have been published for the M.
javanica susceptible cultivars, which are widely grown at present, as
wherever any juveniles have been found, an economic response to
nematicides can be expected.
Pratylenchus
The migratory endoparasitic root-lesion nematodes, Pratylenchus
spp., are widely reported to be found in tobacco fields, but are rarely
of great importance (Table 6.4). The symptoms of Pratylenchus
invasion are brown lesions, which may encircle the root and cause the
cortex to fall off, so giving Pratylenchus damage the name of 'brown
root rot'. The damage can enhance the development of black shank
and other soil-borne diseases (Inagaki & Powell, 1969).
Globodera
The tobacco cyst nematode, Globodera tabacum, has been important
in the shade-grown tobacco of Connecticut since 1951 (Lownsbery &
Lownsbery, 1954; La Mondia, 1995). A sub-species is known to
attack flue-cured tobacco in a limited area of Virginia and neighboring
parts of North Carolina, and a closely related species also reproduces
slowly on burley tobacco, but does not attack flue-cured cultivars, in
Virginia and North Carolina (Miller & Gray, 1972; Miller, 1977;
Komm, et al., 1983; Melton, et al., 1991). Tobacco cyst nematodes are
reported from some other parts of the world (Table 6.4). The attacked
tobacco plants have small roots with the visible cysts attached

 Page 220
Table 6.4 Relative importance of certain plant parasitic nematodes in
some tobacco growing countries.
Country AphelenchoidesDitylenchusGloboderaPratylenchus
Africa
Malagasy 1 1
South Africa 2
Zimbabwe 1
America
Brazil 2 2
Canada 2
Chile 1 1 2
Colombia 1
Mexico 1
Paraguay 1 2
U.S.A. 2 1
Asia and Oceania
Australia 1
China 1 1 2 2
India 1
Korea 1 1 1
Malaysia 2
Pakistan 1 1 2 1
Thailand 1
Vietnam 3
Europe
Albania 1
France 2 2 2
Germany 1 2 2
Greece 1
Hungary 1 1
Italy 2 2 2
Yugoslavia 2 2 1
Mediterranean
countries
Iraq 1
Morocco 2
Turkey 1 1
Key: 1 Minor importance. 2 Moderately important or locally important.
3 Very important.
Adapted from: Survey of Pests and Diseases of Tobacco and Chemicals
Used. CORESTA: Agronomy and Phytopathology Groups (1987).

and can be severely stunted with dark-green leaves. Yield losses can
be high, but at present the distribution is limited and the host range of
these nematodes is restricted to solanaceous plants.
Other Nematodes
Ditylenchus dipsaci, the stem and bulb nematode, is reported from
various countries but only causes yield loss in tobacco in Holland,
France, Germany and Switzerland (Lucas 1975; Valloton and Corbaz,
1976) (Table 6.4). The nematode can invade the lower parts of the
stem causing 'stem break'. This is usually associated with cool, damp
weather and heavy soils and is only of localized importance.
Aphdenchoides species are reported from a few countries (Table 6.4).
However, it is only in a limited area near the French Pyrenees that A.
ritzemabosi has been described as causing leaf blotches on tobacco
similar to those it causes on chrysanthemums (Delon, et al., 1981).
Species of Tylenchorhynchus, Rotylenchulus, and the spiral nematodes
are all reported to feed on tobacco roots, but the damage caused is
generally so slight that they are regarded as being of little economic
impor-

 Page 221
tance (Khan, 1990; Patel & Patel, 1990; Melton & Powell, 1991).
Paratrichodorus and Trichodorus species are vectors of the 'tobacco
rattle' virus, which is reported to cause yield losses in tobacco in parts
of Holland and Germany (Lucas, 1975). Xiphinema and Longidorus
are widespread, and X. americanurn is a relatively efficient vector of
the 'tobacco ringspot' virus which is reported in many countries and
has localized importance (Lucas, 1975).
Control
The extent to which nematode control measures are taken in countries
growing tobacco varies greatly. In some places, such as France and
China, losses due to nematode damage are slight or are of localized
importance only, and little attention is paid to control measures. Such
countries are usually those where tobacco is grown under cool
conditions, often on heavier soil types, and where the root knot
nematode is not widely distributed.
In the USA, Australia, parts of central and southern Africa and other
places where the root knot nematode is widely distributed, the entire
tobacco growing cycle is centered around nematode control. The basic
strategies are to reduce the initial nematode population in the soil and
plant and to reduce the subsequent rate of nematode increase.
However, there are still some countries where nematodes can cause
economic yield loss and little attention is paid to nematode control by
some of the farmers, except maybe at the most basic level.
a
Cultural Control
Some of the earliest control measures practised were the adaptation of
cultivation methods to reduce the nematode population and its effect
on tobacco. The early and efficient destruction of tobacco seedbeds
and roots in the field is an effective way of reducing damage to
subsequent crops by nematodes and other pests (Melton, et al., 1995).
In countries such as Zimbabwe, where the rainfall is in distinct
seasons, it is recommended to plough in the previous crop at the end
of the rainy season. This conserves moisture during the dry season and
also exposes the surface to the sun. The top 50 mm, from which the
tobacco ridge will be made, is often heated to over 36°C and the soil
moisture level goes down to less than 1%, conditions under which
root knot nematode eggs and juveniles are killed (Ferris, 1969).
Preparing the ridge and planting early to take advantage of the lower
nematode populations reduce damage and increase yield (Table 6.5).
In many areas where tobacco is grown by small-scale peasant farmers,
who may find nematicides and nonproductive rotations too expensive,
early deep ploughing, early planting and root destruction are to be
recommended.
Table 6.5 Impact of planting date on
tobacco yield, Zimbabwe, 195354.
Planting date Yield (kg/ha)
8November 1380
19November 1344
3December 1030
17December 461
31December 231
14January 166

A period of bare fallow would reduce the nematode population, but

this practice cannot be recommended because rain and wind would
cause serious erosion if the fallow is to be effective, as it has been
shown that M. javanica can still be recovered from a bare fallow after
4 years (Martin, 1967). Allowing the fields to revert to a natural weed
cover would reduce the risk of erosion, but may not reduce the
nematode problem as many weeds are hosts (Meyer & van Wyk,
1989; Madulu & Trudgill, 1993). The choice of crops to grow in
rotation with tobacco depends on the most important nematodes
present. In much of southern Africa the dominant nematode is M.
javanica, but in most of the rest of the world it is M. incognita and the
situation is made more complicated in many areas where mixtures of
root knot species are found.
In places where M. javanica is the main problem, crops such as cotton
and groundnuts can be grown (Table 6.6) (Madulu, et al., 1994), but
where M. incognita or M. arenaria are found, these crops may not be
satisfactory. In North Carolina and other places where M. incognita is
the dominant nematode, a 4-year rotation with fescue, cereals or
'Rowan' lespedeza is very effective (Melton, et al., 1995). The
sunnhemps can be used to suppress root knot nematodes, although, as
some of them produce a toxin, they cannot be grown in some
countries. Maize, although lightly attacked, is often grown in tobacco
rotations in Zimbabwe, where it is an important food crop, and
provided that it is grown for 2 or more years on its own or as the first
year with other resistant crops, it helps to lower the M. javanica

 Page 222
Table 6.6 Root damage by Meloidogyne
javanica to tobacco after 5 years of various
crops, Zimbabwe, 196061.
Crop Root knot index
(0100)
Maize 18
Groundnuts 2
Cotton 3
Soybeans 32
Sunnhemp (Crotalaria 19
juncea)
Sunnhemp (Crotalaria 1
spectabilis)
Tobacco 66

populations. Certain maize herbicides are very damaging to tobacco,

and a minimum of two seasons must elapse before planting tobacco if
such herbicides are used. Rotation crops can affect P. penetrans
populations where this nematode is a problem (Arsenault, et al.,
1989).
Pasture grasses that are resistant to root knot nematodes are valuable
rotation crops, since they protect the soil from erosion better than row
crops and, if sown densely enough and well maintained, will suppress
weeds which may be nematode hosts. They can also be cropped for
seed or hay. In North Carolina, fescue (Festuca pratensis) is
recommended as a rotation grass (Melton, et al., 1995). In southern
and central Africa, the Ermelo and Umgeni strains of weeping
lovegrass (Eragrostis curvula), Katambora Rhodes grass (Chloris
gayana) and Sabi Panic grass (Panicum maximum) are recommended
(Table 6.7) (York, 1989). To obtain the maximum benefit from these
grasses, it is necessary to grow them for 3 or 4 years and, when
growing a M. javanica susceptible tobacco cultivar, to use a
nematicide (Table 6.8).
Table 6.7 The effects of 3 years of grass on the
yield and Meloidogyne javanica damage to
tobacco grown in unfumigated soil in
Zimbabwe (after Daulton, 1964).
Grass Yield Root knot index
(kg/ha) (0100)
Eragrostis curvula cv 1817 31
Ermelo
Chloris gayana cv 1530 63
Giant
Chloris gayana cv 1966 33
Katambora
Setaria sphacelata cv 1342 80
Kazungula
Panicum maximum cv 1605 50
Sabi

b
Physical Control
Many of the early attempts to control nematode pests of tobacco in
seedbeds or nurseries relied on heating the soil by burning grass and
brushwood on the seedbed area. Though many peasant farmers still
use this as their only method of seedbed control, it is difficult to get
adequate heat penetration to kill nematodes below 150 mm. Burning
can be used to control weeds when a nonherbicidal nematicide is used.
Steam can be used to kill nematodes, weeds, insects and other
pathogens; however, by upsetting the soil bacterial balance, it can lead
to an increase in soil ammonium and to manganese toxicity. If steam
is applied to the soil surface under a cover, effective penetration is
about 300 mm, but greater effectiveness can be obtained using buried
perforated pipes or a steam plough. All steaming methods are slow
and expensive and are only suitable for seedbeds or similar small
areas.
Solarization, the heating of soil covered with plastic by the sun, is an
attractive way of controlling pests and diseases in areas of
uninterrupted sunshine (Patel & Makwana, 1992). However, it is time
consuming, as the soil often needs to be covered for 6 weeks or more.
Also, heat penetration is often not deep enough and consequently the
results are not always successful (Stapleton & De Vay, 1986).
Some degree of nematode control can be obtained where the fields are
flooded, either naturally or when tobacco is grown after paddy rice (de
Guiran, 1970). However, flooding may be needed for at least 100 days
before all the nematodes are killed.
c
Control by Resistance
In the USA many tobacco cultivars have been bred with resistance to
M. incognita races 1 and 3 (Peedin, 1995). These cultivars can be
grown anywhere that has a problem with these two races of M.
incognita and where the type of tobacco produced by these cultivars is
acceptable. The resistance to these two races of M. incognita appears
to be expressed as an inability to establish a feeding site, rather than
an inability to penetrate the root (Schneider, 1991). The source of this
resistance was N. tomentosa (Slana & Stavely, 1981). Other sources of
resistance to race 3 are available from an N. repanda × N. tabacura
cross (Gwynn, et al., 1986) and for races 1 and 4 from a selection of
N. otophora (Arcia & Wernsman, 1983; Reed & Schneider, 1992).
In Zimbabwe where M. javanica is the most

 Page 223
Table 6.8 The effect of length of Eragrostis curvula cv Ermelo ley on
yield and Meloidogyne javanica infestation on tobacco in Zimbabwe
(after Daulton, 1964).
Yield (kg/ha) Root-knot index (0100)
Rotation Not fumigatedFumigatedNot fumigatedFumigated
Continuous 486 1571 93 37
tobacco
1 year grass ley 712 1599 73 21
2 year grass ley 1190 1900 58 18
3 year grass ley 1416 2050 34 11
4 year grass ley 1597 2101 14 7

important nematode, sources of resistance have been found in N.

repanda, some strains of N. longiflora and in some locally grown
primitive N. tabacum types. These lines were crossed to M. incognita
resistant cultivars from the USA, and the advanced resistant lines
were crossed to commercial susceptible cultivars to produce F1
hybrids which have a high level of resistance and produce good
quality leaf (Table 6.9). These hybrids, although producing better
yields and less galling than the susceptible cultivars, do benefit from
the extra protection provided by a reliable nematicide when exposed
to a high nematode population. One of these hybrids has been released
for commercial use and others are near to being released.
No cultivars with resistance specific to Pratylenchus have been
developed, but several of the M. incognita resistant cultivars show
some resistance (Southards & Nusbaum, 1967). Sources of resistance
to the tobacco cyst nematode have been established and a breeding
program started (Baalawy & Fox, 1971; La Mondia, 1991; Herrero, et
al., 1996). There has been no attempt to breed for resistance to any
other nematode parasite of tobacco.
d
Chemical Control
Although many nematicides are banned or under close scrutiny as
they are considered to be a health or environmental hazard, they
continue to be an important tool in tobacco production.
The fumigant nematicides are particularly important in seedbed
management because they are more effective than the nonfumigants.
Methyl bromide, which is one of the halogenated gases claimed to be
responsible for the depletion of the ozone layer, is widely used as a
seedbed fumigant, as it gives excellent control of nematodes, weeds
and other soil-borne pathogens (Table 6.10). However, the parties to
the Montreal Protocol of the UNEP recommended in December 1995
that its use be phased out in the developed countries by 2010,
although some countries, notably the USA, are planning to ban its use
earlier. Its use is to be restricted in the developing countries after
2002. In many countries less expensive fumigants, such as EDB
(ethylene dibromide) and 1,3-D (1,3-dichloropropene), are
recommended for nematode control, but if weed and soil-borne
disease control is needed, it is necessary to burn
Table 6.9 Effects of two nematicides on Meloidogyne javanica resistant
hybrids in a heavily infested site, Zimbabwe, 198586.
Nematicide Rate/ha Yield (kg/ha) Root knot index (0100)
K51 E RK3 RK5 K51E RK3 RK5
(susceptible) (resistant) (susceptible) (resistant)
Untreated 946 2363 2999 90 4 9
EDB 41% m/m1 22.5 l 2693 3761 3868 50 <1 1
45.0 l 4120 3968 4374 42 <1 <1
Aldicarb 1.5 kg 2067 2957 3386 75 1 3
3.0 kg 2075 3220 3598 66 <1 3
1 m/m = proportion by mass.


 Page 224

Table 6.10 Impact of seedbed fumigation on the incidence of weeds,

seedling number and root knot infestation of tobacco seedlings,
Zimbabwe, 1967.
Material Rate Spacing Weeds per Tobacco seedlings Root knot index
m2 per m2 (0100)
Untreated 1281 517 43
Methyl 50 32 624 0
bromide g/m2
EDB 41% 5 ml 35 × 35 1216 624 0
m/m cm
DD/MITC 3 ml 30 × 30 129 613 0
cm

brushwood on the seedbed or apply other chemical treatments. The

mixture of 1,3-D and MITC (methyl isothiocyanate) gives good
nematode control with reasonable weed control. In Sri Lanka and
some other countries, dazomet is used successfully, but in Zimbabwe,
dazomet and metham-sodium, both MITC precursors, have not proved
to be reliable enough to be recommended for use on their own.
In the field, EDB and 1,3-D are the most successful fumigants,
although EDB has been banned in some countries. In the past DD
(1,2-dichloropropane, 1,3-dichloropropene), the forerunner of 1,3-D,
was widely used. In Zimbabwe, as in other countries where the highly
aggressive M. javanica predominates, the fumigants are widely used,
because they provide better control and higher yields than the
nonfumigant nematicides (Tables 6.9 and 6.11). Chloropicrin, either
on its own or in combination with 1,3-D or methyl bromide, is an
effective nematicide and is particularly useful when there is also a risk
of Granville wilt or black shank (Melton, et al., 1995). 1,3-D/MITC
are frequently used in Canada for control of Pratylenchus penetrans
and black root rot (T. basicola) (Tu, et al., 1995).
The nonfumigant nematicides aldicarb, fenamiphos and ethoprophos
are used in the USA, although they are not as effective as the
fumigants (Melton, et al., 1995). The use of aldicarb is restricted in
some states because it has been found in the ground water and has
contaminated some wells. Aldicarb is also used in Zimbabwe, Malawi
and South Africa, but it is not recommended when there is a high root
knot nematode population. Oxamyl, applied as a spray 3 to 5 weeks
after planting, is used in Malawi, South Africa and Zimbabwe as a
supplement to fumigation by extending the period of control.
Fenamiphos can also be used in the same way in Zimbabwe. Many
nematicides are being screened to determine the most effective ones
for control of the tobacco cyst nematode, in particular in Virginia.
The halogenated fumigants, in addition to being phytotoxic, may
increase total nitrogen and alkaloids, lower reducing sugars in the
tobacco leaf and disrupt the normal nitrification processes in the soil
(Tillet, 1964; Elliot, et al., 1972). This is not normally a problem in
the field as the situation has usually returned to normal during the 2 to
3 weeks before the seedlings are planted. However, in the seedbeds, it
is advisable to apply some of the nitrogenous fertilizer in the nitrate
form to overcome the problem.
Care must be used in the safe handling of nematicides, with the use of
protective clothing encouraged. The cured leaf must be analyzed to
ensure that residues are kept to acceptable levels and that
recommended application rates do not lead to unacceptable residues
or to environmental pollution.

Table 6.11 Effects of nematicides on tobacco yield and root knot

nematode damage with a high Meloidogyne javanica
population, Zimbabwe, 198586.
Rate
Material
per plant per ha Yield (kg/ha) Root knot index (0100)
Untreated 1264 88
EDB 41% m/m 3 ml 45 l 3244 32
1,3-D 92% m/m 4 ml 60 l 2958 35
Aldicarb 3 kg 2459 58

 Page 225
e
Biological Control
Worldwide attention is being given to manipulating natural methods
of reducing nematode populations, with the hope that these methods,
which may be less damaging to man and the environment, can, at least
partially, replace chemical control methods.
Research with Paecilomyces lilacinus and Pasteuria penetrans has
shown that they can reduce galling and improve the growth of tobacco
infested with M. incognita (Dube & Smart, 1987; Chen, et al., 1994).
Other fungi, such as Verticillium chlamydosporium, can parasitize the
eggs of Meloidogyne and Globodera and may become useful control
agents (Stirling & Mankau, 1978; Kerry, 1987).
Plants with toxic root exudates, such as the Tagetes species, can be
used to reduce nematode populations in small areas, but unless they
are sown as a dense ground cover to suppress weeds much of the
benefit can be lost (Daulton & Curtis, 1964). Plant extracts, plant
waste and other organic matter can be incorporated into the soil and
will reduce nematode populations (Singh, et al., 1985; Rodriguez-
Kabana, et al., 1990). These soil amendments probably operate
through the breakdown of the organic matter releasing toxic
substances and also by encouraging the growth of antagonistic or
parasitic fungi or by increasing soil enzymatic activity.
f
Summary of Control Measures
There is no doubt that the most effective technique for controlling
nematodes, in particular the root knot nematodes, is by a combination
of all possible methods.
It is essential that the seedbed area be free from all nematode pests,
because if infected seedlings are transplanted into the field, any
control measures used there are valueless. Fumigation with methyl
bromide which kills nematodes, weed seeds, soil insects and soil fungi
is a very effective way to protect the seedlings. If the seedbed area is
not used every year, it should be sown to a nematode-resistant crop. In
the field, tobacco should not be grown in the same field frequently; a
break of 3 or 4 years of nematode-resistant crops, preferably a grass,
will lower the nematode population throughout the whole field, thus
making any chemical control more effective (Daulton, 1964), (Table
6.8).
Many cultivars resistant to M. incognita races 1 and 3 and some
resistant to M. javanica are available, and where these nematodes are
the main problem, the resistant cultivars are very valuable. In many
situations, the use of a good rotation and a resistant cultivar may
provide enough control, but often the addition of a nematicide is
needed to realize the full potential of the resistant cultivar. If the
nematode population is high, a fumigant will give better control than
an organophosphate or carbamate nematicide. Multi-purpose
fumigants, such as 1,3-D + chloropicrin or 1,3-D/MITC, should be
used in fields with nematodes and Granville wilt or other soil-borne
disease. However, the choice of nematicide may be limited by cost or
by restrictions on the sale of certain nemaficides.
Biological control measures using antagonistic or parasitic fungi and
Pasteuria penetrans are attractive, but not enough is known about
such organisms or their mass production to know whether they can be
commercially viable. Advanced plant breeding techniques and genetic
engineering offer the prospect of new cultivars with multiple
resistance to nematodes, disease organisms and insects and with
tolerance to presently phytotoxic chemicals.
With the increasing concern over the use of toxic chemicals, the future
of nematode control in tobacco growing lies in breeding resistant
cultivars, the intelligent use of cultural and cropping techniques and
the use of minimal amounts of chemicals, possibly backed up by
biological agents.
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 Page 228

Chapter 7
Tobacco Insect Pests
7A
Insects and Their Management in Tobacco Production
B.W. Blair
Tobacco Research Board
Harare, Zimbabwe
Introduction
Insects and their control have been~ the focus of numerous
publications over several decades from agricultural experiment
stations, for example Kuitert and Tissot (1956). In this section only
those insect pests of tobacco that are considered to be important in
several of the tobacco-producing areas have been included. There will
undoubtedly be pests of importance in particular countries or regions
that are not mentioned here, and interested readers are advised to
obtain such information from relevant sources in those areas. No
specific insecticides are mentioned because for a particular pest there
may be differences in the insecticides registered or used in each
country. Furthermore, since new recommendations are continually
being made, any mention of specifics would soon be outdated.
Information on actual recommendations is available in various forms
from most tobacco-producing countries, for example from North
Carolina Cooperative Extension Service in the USA (Southern, 1995),
Tobacco Research Board in Zimbabwe (Anon., 1990), Central
Tobacco Research Institute in India (Chari & Nagarajan, 1994), and
National Tobacco Administration in the Philippines (Xuan, 1989).
For each insect pest mentioned, information is given on its broad
geographical distribution, on the pest's life cycle, descriptions of the
various stages in the cycle and the damage to tobacco, and there are
general comments 1 on the management of the pest.
Insect Life Cycles
For readers who do not have a knowledge of entomology, there are
two types of life cycles in the insect kingdom with some variation in
each. As insects have an external skeleton (exoskeleton), insects at
immature stages need to shed this periodically to allow for growth
into a new soft exoskeleton and before this hardens. The stages
between moults are known as instars.
The incomplete (hemimetabolous) cycle consists of an egg, from
which a nymph hatches and which resembles the adult but does not
have fully developed wings or reproductive organs. The nymph
moults a number of times before it becomes fully grown and develops
rudiments of wings and reproductive organs (Fig. 7.1).
The complete (holometabolous) cycle consists of an

Fig. 7.1
Heminetabolous (incomplete) insect life cycle.

 Page 229
egg, from which hatches a larva that moults a number of times before
it changes into a pupa. The pupa is a resting stage and does not feed.
Within the pupa the larva undergoes tremendous change to form the
adult. The larva and pupa do not resemble the adult, though the form
of the adult can sometimes be seen through the exoskeleton of the
pupa (Fig. 7.2). The larvae of some orders of insects are sometimes
commonly known by other names, such as caterpillars (butterflies and
moths), grubs (beetles) and maggots (flies).

Fig. 7.2
Holometabolous (complete) insect life cycle.
As insects are cold-blooded (poikilothermic), their rate of
development is largely dependent on ambient temperature. During
unfavorable periods the insects may enter a type of hibernation
(diapause) so as to survive. Besides temperature, the availability and
nutritive value of food, moisture and daylength may also affect insect
development.
In some insects and/or in some conditions there may be no sexual
reproduction as there are no males. The females are able to produce
viable eggs by a type of asexual reproduction known as
parthenogenesis.
Chemical Insecticides
As a general principle, chemical insecticide control of insect pests
should only be considered if they are causing economic damage, that
is, once the economic injury level has been reached. In crops where
seeds or fruit are the harvested product, a fair amount of leaf damage
can occur before there is an economic loss of yield. This means that
the insect injury thresholds can be relatively high. In tobacco leaf
production, however, it is the leaf that is harvested so any loss of leaf
has a direct effect on yield and any damage could also reduce quality;
in other words, the economic pest injury levels in tobacco leaf
production are usually lower than for the same or similar pests in
other crops.
Before any control is used, it is important first to identify the pest
(publications with color plates are available in some regions to assist
in identification) (Reich, 1985; Xuan, 1989; Chari & Nagarajan, 1994;
Anon, 1995a), since any one insecticide or other form of control
measure will not be effective in controlling all insect pests. The
labeled or recommended rates, methods and timing of insecticide
application should be followed to ensure that:
lowest effective doses are applied;
insecticide residues in the cured tobacco are minimal;
pest populations do not develop resistance by being exposed to
excessive rates or to unnecessary applications;
the recommended application method is used so that the insecticide
reaches its intended target;
pests are controlled before economic damage occurs and before it
becomes difficult to reduce pest numbers below damaging thresholds,
either because the insect has grown and is more difficult to control or
because of sheer numbers.
The continual use of insecticides from the same chemical group (such
as organophosphates, carbamates and synthetic pyrethroids) may lead
to pests developing resistance to that group, and rotation of groups or
limiting their use will assist in prolonging the effective lifespan of the
pesticides (Blair, 1986). It is important to retain the present
insecticides for tobacco pests because the cost of developing,
evaluating their efficacy and registering new ones is now very
expensive, and the manufacturers have to balance these costs with
potential sales. Because tobacco is not a major crop in terms of
pesticide usage worldwide, manufacturers are now generally less
inclined to register new products for controlling tobacco pests.
General guidelines for the selection, application and handling of
agrochemicals in leaf tobacco production have been published by the
Cooperation Center for Scientific Research Relative to Tobacco
(CORESTA) (Anon, 1993). There is a useful guideline dealing with
the principles of scouting for pests and of treatment thresholds
(Linker, et al., 1989).

 Page 230
Integrated Insect Management
The concept of employing as many methods (chemical, biological,
cultural, resistant cultivars and legislative) as possible to control a pest
is termed integrated pest management. Although effective control may
not be achieved by using any one of the components, the combination
of methods will usually result in reducing the damage to near or below
economic injury levels; the more methods that can be used, the better
the control. Such an integrated approach also allows a more
sustainable (i.e. economically viable, socially acceptable and
environmentally healthy) state to be established for tobacco leaf
production in the long term. The objective of pest management is to
control pests at the least cost and with the least quantity of pesticide:
the concept of complete annihilation is rarely achievable and should
not be the goal. An important aspect in implementing integrated
control is planning to effectively combine each component over a
period of time (not only for the current season but for subsequent
ones).
Careful consideration in the choice of insecticides for an integrated
program is required to minimize deleterious effects on beneficial
natural control agents. In general, soil-applied insecticides have
minimal effects on beneficial insects that frequent the foliage, whereas
long-acting and broad-spectrum insecticides applied to the foliage
have a very detrimental effect. Spot treatment, determined by pest
scouting, of only the areas where a pest is present at or above a
damaging level is much more desirable than blanket applications to
the entire crop, as this allows the beneficials to survive in the
untreated areas.
Economics of Insect Damage and Control
Estimates have been made for the cost of disease losses in tobacco
(for example, Anon, 1995b), but only a few such estimates have been
done for insect damage. In South Carolina, field losses due to insects
(Manley, 1995) were estimated to be 2.63% in 1993 and 0.78% in
1994. The latter damage was valued at US$1.38 million, whereas the
estimated cost of controlling the insects was US$2.5 million. This
example would seem to be uneconomic control. However, the cost of
control can also be interpreted as having been economic since the
control measures limited the degree of insect damage and, by
extrapolation, the total cost of insect damage and control was US$3.88
million for tobacco produced on about 22 000 ha.
Simply, insect injury results either in the death of a plant or the
destruction of some of its tissues. Death is more common in newly
planted tobacco attacked by soil pests. Although there is some
compensatory growth by plants on either side of a missing one, the
quality of tobacco is reduced and, because of the resultant uneven
plant growth, any compensatory benefits are nullified.
The consequences of plant tissues being destroyed or disfigured are
complex, and the compensation within and between plants is more
difficult to measure and define. Because laceworm, Spodoptera
littoralis, feeds on fully expanded leaves of mature plants, results
from laboratory feeding studies (Shaw, 1976) can probably be directly
applied to a field situation. Studies showed that each laceworm
consumed the equivalent of 800 mg flue-cured tobacco and that an
infestation density of 25 laceworms per 100 plants justified the cost of
control. On the other hand, budworms are more destructive on young
tobacco when they remove tissue from unexpanded leaves in the bud,
because this eventually represents large missing areas of the mature
leaves. From field studies it was shown (Shaw, 1981) to be profitable
to save 0.23% of plants from early budworm attack. Once stem
elongation occurs, however, the plant compensates completely for any
subsequent budworm damage.
When insects transmit debilitating virus diseases to young plants,
there is huge loss of leaf yield and quality. In two field experiments, it
was shown that an infection of potato virus Y reduced monetary
returns by 30% and 42%. Early infection of bushy top virus reduced
returns by 93% and 85%, whereas a late infection caused 40% and
62% (Shaw, 1981) reduction. With such high losses, it is obvious that
it would be an economic advantage to control insect vectors.
Soil Pests
Apart from the direct damage to tobacco as a result of the feeding
activities of soil pests, the damaged subterranean parts of the plant
indirectly allow easy entry of soil-borne pathogens. These pathogens
can cause serious diseases such as sore shin, bacterial wilt, fusarium
wilt and black shank (see Chapter 6A of this monograph).
a
Cutworms
These are larvae or caterpillars of night-flying moths that attack the
stems of tobacco below, at or above soil

 Page 231
level. The species include Agrotis ipsilon (cosmopolitan) (Anon,
1952), A. segetum (Europe, Africa, Asia) (Anon, 1952), Euxoa
messoria (North America) (Akehurst, 1981), Feltia subterranea
(Americas) (Akehurst, 1981) and Spodoptera litura (Asia,
Australasia) (Anon, 1952).
The adults of all species are inconspicuous moths having grayish-
brown forewings with various patterns, a wingspan of 40 to 45 mm
and creamy-colored hind wings. The larvae (cutworms) are greasy,
brownish or grayish to black caterpillars which are up to 40 mm long
when fully grown (Fig. 7.3). The early larval instars feed on tender
and prostrate foliage during the day, while the third instar and later
instars rest in the soil during the day and emerge at night to feed on
stems and foliage. The pupae are plump, reddish-brown at first but
darken later and are found in the soil within a cocoon of soil particles.

Fig. 7.3
Black cutworm and moth, Agrotis ipilon.
Cutworms are indiscriminate and wasteful feeders and will attack any
plant that is not too woody. They mainly cut the stems of seedlings or
transplants in seedbeds or soon after transplanting into the field,
directly causing a reduction in plant stand or indirectly by weakening
stems. The larvae of S. litura may also feed extensively on leaves of
young plants, and only the mid-rib remains.
The nocturnal moths lay eggs singly or in clusters on low-lying
vegetation, clumps of soil and preferentially on fibrous plant material
on the soil surface. A female can lay between 600 and 800 eggs and
these hatch in 4 to 13 days. The larval stage takes from 3 to 11 weeks
to complete, after which pupation takes place in the soil. The pupal
stage lasts for 10 to 14 days (Blair, 1976). Depending on the ambient
temperature and availability of food, there can be from one to five
generations per year (Akehurst, 1981; Blair, 1982).
Cutworms are generally controlled by the application of contact
insecticides applied to the soil around the base of plants or by
broadcast incorporated application. The treatment threshold for field
infestations in most countries is when 5% of small plants are injured
or killed up to 3 weeks after transplanting. As cutworms require green
plant material to survive, a cultural method of control is to starve any
resident population by ensuring a bare fallow field for at least a month
prior to transplanting (Blair, 1975). Regular scouting for the presence
of cutworms and damage in seedbeds is desirable because of
potentially high losses that can occur in densely sown seedbeds.
b
False Wireworms
These are the larvae of surface beetles which feed on roots,
underground parts of the stem and on humus. Plants with damaged
roots wilt rapidly, while plants with damaged stems usually die.
Larval attacks are more severe in reverted or virgin fields, because
grasses are their natural food. The adult beetles may also feed on
folded tobacco leaves in the bud.
The larvae are yellow-brown, smooth, elongate and have a tough
shiny exoskeleton (Fig. 7.4). The fully grown, lesser false wireworms
range in length from 8 to 38 mm, while those of larger false
wireworms vary from 50 to 75 mm in length. The main genera
attacking tobacco are Gonocephalum (Africa, Asia, Australia) (Hill,
1987), Trachynotus (Africa, Caribbean) (Büttiker, 1994) and
Psarnmodes (Africa) (Büttiker, 1994). Gonocephalum adults are
brown to black in color, usually coated with a layer of dust,
rectangular and flattened, 10 mm long and 5 mm wide, and with the
wing covers having longitudinal ridges. Adults of Trachynotus are
dull black, though usually coated with dust, and are about 15 mm
long. The margins of the prothorax are

Fig. 7.4
alse wireworm, Tenebrioniade.

 Page 232
angled and there are three prominent ridges on the fused wing covers.
Psarnmodes adults are shiny black, smooth with a globular prothorax
and abdomen, and are about 30 mm long.
Females of Psammodes lay eggs in soil over a period of 1 to 2 months
after the onset of rains and they hatch in 5 to 10 days. The larvae
(larger false wireworms) develop rapidly before the onset of the dry
season, then they burrow deeper into the soil and stop feeding until
the next rains. Larval development takes 2 years, followed by
pupation in an earthen cell and the adults emerge after the first heavy
rains. The life cycle takes 3 years to complete.
Trachynotus and Gonocephalum females lay eggs towards the end of
the rains and these hatch in 7 to 10 days. The larvae (lesser false
wireworms) feed through the dry season and are fully grown in 7 to 8
months. Pupation in a flimsy earthen cell occurs towards the end of
the rains. There is normally one generation a year, but occasionally
the fully grown larvae may not pupate until the following year.
Pre-sowing or pre-planting fumigation of soil effectively reduces false
wireworm infestations. The addition of insecticides to the planting
hole at the time of transplanting also provides protection of the roots
for a few weeks during crop establishment.
c
White Grubs
These are the larvae of beetles, and there are numerous species that
damage tobacco roots in many parts of the world. The larvae are up to
50 mm long, C-shaped, dirty white or yellowish (hence the name
white grub) with a brown head and prominent mandibles, and a
swollen abdomen (Fig. 7.5). The adults vary in size from 10 to 35 mm
long, with relatively hard wing covers; they are plump and brown,
yellowy or gray.
Adults emerge after the first soaking rains and swarm around trees at
dusk and are attracted to light. The female lays about 50 eggs over 2
to 3 weeks at depths of 100 to 200 mm in soil. Plowed land is
attractive to egg-laying females, as it is easier for them to burrow.
Eggs hatch in about 18 days and the larval period spans about 6
months. As the dry season approaches, the larvae burrow deeper into
the soil and then pupate 3 to 4 months later in earthen cells. The pupal
period lasts between 14 and 28 days, and the adults emerge when
moisture again contacts the pupal cell.
The white grubs feed on organic matter and at the root crown; the
latter damage is clean cut and results in

Fig. 7.5
White grub, Scarabaeidae.
reduced growth and wilting; young transplants may be killed. The
adult chafers can also severely defoliate tobacco, particularly the
upper leaves and buds of plants on the edge of a field.
Control of white grubs involves incorporation of soil insecticides prior
to transplanting as a preventive treatment, but effective control is
rarely achieved. The incidence of white grubs is reduced when a soil
fumigant is used to control nematodes in fields.
d
Wireworms
These are the larvae of click beetles that attack roots, burrow into the
base of stems and feed on organic matter. There are many species that
feed on tobacco, but the most common are Limonius (North America)
(Akehurst, 1981), Agriotes (Americas, Asia, Europe), Melanotus
(North America, Europe, Asia), Conoderus (Americas, Australasia)
(Akehurst, 1981) and Hemicrepiclius (North America, Europe).
The adult click beetles are generally sombercolored, tapered and
flattish with comb-or saw-like antennae and a peg on the underside of
the thorax which can be forced with a click into a cavity. If a beetle
should land on its back, a sudden release of the peg flicks the beetle
into the air and onto its legs. The larvae are cylindrical, hard-skinned,
waxy yellow or brown in color, with two appendages at the rear,

 Page 233
and are 13 to 20 mm long (Fig. 7.6). The larval period varies from 1 to
3 years according to the species. Pupation takes place in an earthen
cell in the soil, with the pupal period being about 3 weeks.

Fig. 7.6
Wireworm, Elateridae.
By damaging the roots and tunneling into the base of the stem of
newly transplanted tobacco, the wireworms cause the plants to wilt
and die. The severity of attack is less when there are high levels of
organic matter in the soil.
Control of these pests is difficult and usually involves the
incorporation of insecticides in the soil as a pre-plant treatment. It is
difficult to apply post-plant remedial treatments.
Above-Ground Pests
a
Aphids
Aphids occur as winged or wingless forms (Fig. 7.7), the most
familiar being the wingless. The main species on tobacco are the
green peach aphid, Myzus persicae (cosmopolitan) (Anon, 1952), and
the red tobacco aphid, M. nicotianae (Americas, Africa, Southern
Europe, Middle East and South-East Asia) (Blackman, 1987). The
latter aphid was first thought to be a red morph of M. persicae which
has now displaced the green morph in the USA, but Blackman (1987)
maintains that it is a separate species. Aphids suck sap and nutrients
from the plant, which stunts growth and causes distortion. While
feeding they produce a sugary secretion (honeydew) on which sooty
mold grows and discolors the leaf. Aphids are more important as
vectors of virus diseases (see virus diseases in Chapter 6B of this
monograph) of tobacco in many parts of the world. Heavy aphid
infestations and virus infections cause reductions in both yield and
quality of tobacco.
Aphids are soft-bodied, oval and 1.5 to 2.5 mm long with a pair of
conspicuous tubes (cornicles) near the tip of the abdomen. The
wingless forms are yellowy-green to pale-green, and some may have a
reddish color (green peach aphid) or most may be reddish-brown
(tobacco aphid). The winged forms have darker heads

Fig. 7.7
Green peach aphid, Myzus persicae, (a) winged
form, (b) wingless form.
and thorax, dark markings on the abdomen and dark cornicles. They
occur in colonies on the underside of leaves, suckers and buds.
In almost all tobacco-producing areas M. persicae does not reproduce
sexually as no males are present, while in the more temperate areas
there may be both sexual and asexual reproduction. There is no sexual
reproduction in M. nicotianae (Blackman, 1987). In asexual
reproduction both winged and wingless females can each produce
about 80 offspring. Embryonic development begins before the
mother's birth, in the body of the grandmother whose food intake
sustains three generations of about 6000 individuals. This peculiar
reproduction, together with a short life cycle of 7 to 10 days, is the
reason for the rapid build-up of aphid populations in a crop. Winged
females are formed when there is overcrowding or when there is a
change in the nutritive quality of the tobacco. The winged aphids are
able to migrate to new sources of food. Where aphids reproduce
sexually, males are formed during the autumn and cold-hardy
overwintering eggs are produced on the primary host (peach in the
case of M. persicae).

 Page 234
The thresholds at which to apply aphid control vary according to the
stage of crop development and whether virus diseases are endemic or
not. The thresholds up to topping are very low in areas where aphid-
transmitted viruses are endemic, and routine aphid control treatments
are usually advised (Anon, 1990). The treatment threshold in North
Carolina (Southern, 1995), where aphid-transmitted viruses are not
endemic, is when 10% or more plants have at least 50 aphids on an
upper leaf before topping; after topping it is when 20% of the plants
are infested (Southern, 1995). Chemical control normally involves the
application of systemic insecticides, as foliar sprays, transplant water
treatments or as soil-applied granules. There are many natural enemies
of aphids (parasitic wasps, ladybird beetles and larvae, plus hover-fly
larvae), and the use of soil-applied systemic insecticides is preferred
as there is minimal harm to these natural control agents. In Zimbabwe
there is legislation that enforces a 'dead' period (when no tobacco is
growing) to minimize the carry-over of virusbearing aphids from one
season to the next. The legislation prohibits transplanting at a time of
high aphid populations (Blair, 1994).
b
Budworms
Budworms are the caterpillars of moths that have gray to gray-brown
forewings with wavy transverse lines, beige hind wings with darker
bands near the edge and a wingspan of about 35 mm. The younger
larvae feed on tender tissues of the buds and older larvae may feed on
the expanding leaves. In seed crops they will bore into the flowers and
developing seed capsules. There are three main species that damage
tobacco: Heliothis virescens, Helicoverpa (Heliothis) zea (Americas)
(Anon, 1952), and Helicoverpa (Heliothis) armigera (Africa,
Southern Europe, Middle East, Asia, Australasia) (Anon, 1952), but in
all cases tobacco does not seem to be an ideal food plant.
The yellowy-white eggs are ribbed and domeshaped. The older larvae
vary in color from shades of green to pink, brown or almost black but
can be recognized by having three dark lines running down the middle
and sides of the body which are separated by two broad pale stripes
(Fig. 7.8). The larvae are 40 mm long when fully grown. The pupae
are mahogany brown and are found in the soil.
The female can lay several hundred eggs which are laid singly on
leaves, stalks or buds. The eggs hatch in 3 to 8 days, and the newly
hatched larvae eat their egg shells as their first meal before moving
towards the bud. The larval period is about 18 days in summer or

Fig. 7.8
Budworm, Helicoverpa (Heliothis) armigera.
up to 50 days in winter, and the pupal period varies from 14 days in
summer to several weeks in winter. There can be several generations
per year.
Tobacco genotypes that have fewer glandular hairs (trichomes) and,
therefore, fewer sticky surfaces than normal are more susceptible to
budworm attack. Control of budworms can be by chemical
insecticides and biological agents, such as the bacterium Bacillus
thuringiensis. The treatment threshold in North Carolina (Southern,
1995) is when 10% of the plants are infested and in Zimbabwe the
threshold is 10 budworms per 100 plants (Anon, 1990).
c
Flea Beetles
These are brown beetles with black markings, about 1.5 mm long,
with fine indentations on their wing covers and with yellowy antennae
and legs. Three species of Epitrix (Americas) (Akehurst, 1981) feed
on tobacco (Fig. 7.9) in seedbeds and in the field, producing small
round holes that give a shot-hole appearance. Although the beetles are
small, serious damage can occur when large numbers feed on newly
transplanted tobacco or on lower leaves of larger plants. The larvae
also feed on and burrow into roots of tobacco seedlings, causing them
to be unthrifty or in some cases to die.
Eggs are laid in shaded areas on moist soil beneath tobacco plants.
The larvae are slender and white with brown heads and are found in
the soil around roots or burrowing into the roots. The larval period
lasts 4 to 5 weeks, and pupation occurs in the soil. There are two to
four generations a year (Rabb, et al., 1959).
The treatment threshold in North Carolina (Southern, 1995) is when
there are four or more beetles per plant on small plants, and on large
plants when there are 60 or more beetles per plant or when the lower
leaves look lacy.

 Page 235

Fig. 7.9
Flea beetles and larva, Epitrix spp.
d
Grasshoppers
There are numerous species of both short-and longhorned
grasshoppers that will feed on tobacco. Examples of species are
Melanopus (Americas) (Anon, 1982), Phaneroptera (Asia, Europe),
Zonocerus, Ruspolia and Phymateus (Africa) (Anon, 1982). All the
grasshoppers have enlarged hind legs and are 25 to 85 mm long. They
vary in color from green to brown or black with areas of bright colors.
They feed by chewing large irregular holes in leaves which can result
in complete defoliation of young plants. As the natural habitat of these
insects is grassland and bush, the damage is first seen on the edges of
tobacco fields and seedbed sites.
Depending on the species, eggs are laid either in soil or in slits in
plant tissue. The nymphs may be gregarious for a period after
hatching, and they feed on foliage while undergoing four to five
moults before becoming adults. There may be one to three generations
a year, and in some species a generation may take 2 years because the
eggs take a year to hatch.
If there are small numbers of grasshoppers in patches on the edges of
fields, it may be possible to collect them by hand. Sprays or dusts of a
contact insecticide may also be used to control grasshoppers,
particularly in seedbeds.
e
Hornworms
These are caterpillars of hawk moths and are characterized by a fleshy
horn at the tip of the abdomen. The main species feeding on tobacco
are Manduca sexta and M. quinquemaculata (Americas) (Hill, 1987).
After hatching the larvae are pale green and about 6 mm long,
becoming bright green or brownish and attaining a length of 75 to 90
mm; those of M. sexta have seven white diagonal stripes on each side
and a curved red horn, while those of M. quinquemaculata have eight
white V-shaped marks on each side with a blue-black horn (Fig. 7.10).
The adult moths are 45 to 50 mm long and the wings (span 80 to 100
mm) are held in a delta shape when at rest; coloration is grayish
brown with lighter wavy markings on the wings and orange spots on
the abdomen. The eggs are spherical, about 1.5 mm in diameter,
greenish when laid and becoming almost white later.
 Fig. 7.10
Hornworms, (a) Manduca sexta, (b) M.
quinquemaculata.
The young larvae chew small irregular holes in leaves, while the
larger ones feed voraciously from the leaf margins, leaving only the
mid-ribs and larger veins.
The female moth lays about 300 eggs singly on the underside of
leaves. The eggs hatch in 3 to 6 days, and the larval period takes 25 to
30 days to complete. The fully grown larva burrows into the soil to
pupate and in summer the adult moth emerges about 2 to 3 weeks
later; the pupal period is longer in winter.
The treatment threshold in North Carolina

 Page 236
(Southern, 1995) is when there is one hornworm per 10 plants or five
hornworms per 10 plants if they have white silken cocoons of wasp
parasites attached to their bodies.
f
Laceworm
This is the caterpillar of the night-flying moth Spodoptera littoralis
(Southern Europe, Africa) (Anon, 1952), which feeds on leaf tissues
between the veins so that damaged leaves have a lacy appearance. It
causes serious defoliation to the lower leaves of topped plants. Also, a
related species, S. litura (see (a) under Soil pests, this chapter) can
defoliate tobacco.
The moth has a gray-brown body, is 15 to 20 mm long and has a
wingspan of 35 mm. The forewings of the female are gray-brown with
paler lines along the veins, whereas in the males there are bluish areas
at the bases and tips. The hind wings are white with darker veins and
gray margins. The eggs are 0.6 mm in diameter, spherical, slightly
flattened and white or yellow. Fullygrown larvae are up to 45 mm
long and their color varies from yellowy-white to bluish-gray and
grayish-brown. On either side of the mid-line of the abdomen there
are two rows of triangular black markings, with those on the first and
eighth segments being larger than the others. On the side of the
abdomen there are alternating dark and light longitudinal bands.
The female can lay up to 1500 eggs in clusters of about 300 on the
underside of leaves and they are covered with gray-brown scales from
the female's abdomen. The eggs hatch in 3 to 4 days and the young
larvae are gregarious. As the larvae grow they disperse; the older ones
tend to leave the plant to shelter under debris and clods of soil during
the day and return to feed at night or early morning. The larval period
lasts for 15 to 20 days, then the larvae enter the soil to pupate. The
adults emerge from the pupae in about a week and live for about 10
days. There are several generations a year (Blair, 1990).
Control of this pest is usually obtained by spraying chemical or
biological insecticides to the lower half of the plant. The treatment
threshold in Zimbabwe is when there are eight or more laceworms per
100 plants. There are many wasp and fly parasites of laceworms, and
there are viral and bacterial diseases of the larvae which assist in
natural control.
g
Loopers
These are caterpillars that move by drawing up the abdomen in a loop
as they only have prolegs on the last three abdominal segments (most
caterpillars have prolegs on five segments). There are two main
species that feed on tobacco (Fig. 7.11), Trichoplusia ni (Americas)
(Anon, 1952) and Thysanoplusia orichalcea (Africa, Middle East)
(Anon, 1952). Moths of both species have mottled brown forewings
and paler straw-colored or grayish-brown hind wings which are darker
near the edge. On the forewing of T. ni are two silvery spots near the
center, whereas T. orichalcea has a prominent shiny gold wedge. The
eggs are pale-green, round with ridges and about 0.6 mm in diameter.
The larvae of both species are pale-green with fine white and darker
green lines along the body, and they attain a length of about 35 mm.
The younger larvae feed on the underside of leaves making windows,
whereas older ones feed on the entire leaf but tend not to eat the veins.
 Fig. 7.11
Looper caterpillar, pupa and moth,
Trichoplusia ni.
Eggs are laid singly on the underside of leaves or on stems, and these
hatch in 3 to 4 days into whitish larvae with black spots. They soon
become green and in about 15 days they are fully grown. A white
silken cocoon is spun on the underside of leaves in which they pupate.
The pupa is dark brown and the adult emerges from it in 7 to 10 days.
Several generations are possible in a year.
Loopers are inclined to be sporadic pests and chemical control is used
only when there are sufficient numbers to inflict economic damage.

 Page 237
h
Stink Bugs
These sap-sucking insects are shield-shaped, 12 to 15 mm long and 5
to 8 mm wide (Fig. 7.12). Stink bugs, also known as shield bugs,
refers to a group of insects that may be from different genera and
species from the family Penpatomidae (order Hemiptera). They are
either green, such as Nezara viridula (cosmopolitan) (Anon, 1952)
and Acrosternum hilare (Americas) (Akehurst, 1981) or are brown
such as Euschistus servus (Americas) (Akehurst, 1981) in color. Eggs
are barrelshaped, 1.2 × 0.75 mm; they are white when first laid and
turn pinkish later. The nymphs are smaller versions of the adults and
are gregarious soon after hatching. Both adults and nymphs feed on
plant sap, usually from veins of tender leaves, and they probably
inject a toxic saliva as the area around the feeding site wilts. If
damage occurs during hot and dry weather, the wilted area turns
black, but if the weather is cooler and humid the wilted parts usually
recover. In tobacco seed crops the bugs will also feed on flowers,
causing them to abort, or they will feed on the seed capsules, causing
the seeds to shrivel.
A female can lay up to 300 eggs in batches of about 50, stuck together
in rafts on the underside of leaves. On hatching the nymphs do not
feed but congregate near the eggs. After moulting they disperse and
start
 Fig. 7.12
Stink bug, Nezara viridula.
feeding on plant sap. There are five nymphal instars which take about
2 months to complete; there are about three generations a year, with
the adults hibernating during the winter.
Stink bugs are sporadic pests, but when they appear in a crop they are
usually in large numbers and can inflict serious damage in hot, dry
conditions. Chemical control is used as a remedial treatment.
i
Thrips
Thrips are torpedo-shaped insects, 1.0 to 1.5 mm long, and are pale-
brown to black with darker bands across the abdomen (Fig. 7.13).
Thrips tabaci (cosmopolitan) (Anon, 1952) and species of
Frankliniella are widespread pests of tobacco (Hill, 1987). The adults
have two pairs of finely fringed wings that are folded along the body
when at rest. The nymphs are whitish to paleyellow but have no
wings. Thrips characteristically flex the abdomen over the body and
have a globule of dark excrement at the tip. The small eggs are bean-
shaped and laid in slits in leaves. They feed by rasping the surface of
leaves, particularly in the depression of veins, which results in a
silvery appearance. Thrips are vectors of tomato spotted wilt virus
(see virus diseases in Chapter 6B of this monograph); the virus can
only be spread if the nymphs first feed on an infected plant.

Fig. 7.13
Thrips.
Males are usually scarce, but females are able to produce viable eggs
without fertilization (parthenogenesis), in which case all the progeny
are females. When there is bisexual mating, all the progeny are males.
The female lays about 50 eggs in leaf tissue and

 Page 238
these hatch in 1 week. The nymphs moult twice in about 5 days; the
third instar shows the first signs of wings and stops feeding
(sometimes called a pre-pupa). Pupation occurs in the soil and the
pupae have longer wing buds than the pre-pupae and antennae are
fixed over the head. Adults emerge in about 4 days. The cycle takes
about 3 weeks and several generations are possible in a year. Warm,
dry weather favors the rapid build-up of populations.
Tobacco does not seem to be an ideal host for thrips as they do not
survive for long before being trapped on the sticky trichomes. They
only breed on tobacco once it has flowered.
Remedial chemical control is applied when damage is first seen,
mainly to curtail the spread of tomato spotted wilt virus.
j
Tobacco Leaf-Miner or Splitworm
This is a small caterpillar that tunnels in leaf tissue and sometimes
stems, leaving a transparent area (mines). If the tobacco is small or
growing slowly, the leaf-miners move rapidly up the plant to attack
the heart, causing stunting or death of the plant. There is one
cosmopolitan (Anon, 1952) species, Phthorimaea operculella, which
is also known as splitworm and potato tuber moth (Fig. 7.14).
The adult moth is small, torpedo-shaped, graybrown with a wingspan
of about 12 mm and is active at night. The tiny, pearly-white eggs are
laid singly or in
 Fig. 7.14
Tobacco leaf-miner/splitworm, pupa and moth,
Phthorimaea operculella.
clusters of five or six on soil or foliage. The fully grown larvae are 12
mm long, dirty white or gray in color or sometimes pinkish or pale-
green prior to pupation.
The eggs take about a week to hatch and the newlyhatched larvae
wander over the leaves for a while before tunneling into the plant. The
larva becomes fully grown in 10 to 20 days, then it spins a silken
cocoon in which particles of soil or leaf fragments are incorporated
and in which it pupates. In warmer months each generation lasts about
4 weeks, but at lower temperatures the cycle may take 10 weeks.
As infestations can build up rapidly when temperatures increase after
winter, it is usually necessary to apply chemical control as either a
preventive or as a corrective treatment. There are a number of
parasitic wasps (such as the South American Copidosoma
uruguayensis and Apanteles subandinus which have been released in
many countries) that are effective in controling the pest after the post-
winter flush. As this pest feeds on other solanaceous plants, carry-over
from one season to the next can be reduced by removing solanaceous
weeds from tobacco fields and their borders.
k
Tobacco Slug
This is a slug-like, slimy globular larva of the beetle Oulema bilineata
(South America, Africa) that feeds on tobacco leaves. The first two
larval instars feed superficially on the underside of leaves and the
damage shows on the upper surface as irregular pale-green marks or
blotches. Older larvae penetrate the leaves to make holes and they are
capable of destroying whole leaves.
The adult beetle is rectangular, 6 to 7 mm long and usually black with
a yellow longitudinal stripe on each wing cover. The prothorax is also
yellow with two black spots. The eggs are cylindrical and 0.8 mm
long × 0.3 mm in diameter. The globular larva has a blackish head and
a tapering grayish abdomen. The slimy body is usually covered with
feces.
Females lay up to 1000 eggs in brown packets, containing 15 to 30 in
each, which are stuck to the underside of leaves. Eggs hatch in about 1
week and the larvae take about 2 weeks to become fully grown. These
drop to the soil to pupate and adults emerge about two weeks later.
The adults of the first two summer generations live for about 2
months. Soon after the adults of the third summer generation start to
lay eggs, they penetrate the soil to about 100 mm to overwinter and
emerge at the onset of rains to continue egg laying.
As outbreaks of this pest are sporadic, it is normal to spray
insecticides as a corrective treatment. Alternative

 Page 239
solanaceous food plants act as a reservoir for the tobacco slug. Growth
of these alternate hosts should be discouraged when possible.
l
Tobacco Stem Borer
This is a small, white-bodied, brown-headed caterpillar of a moth
similar to that of the tobacco leaf-miner (see (j) Tobacco leaf-miner or
splitworm, this chapter). There is one species, Scrobipalpa heliopa
(Africa, Middle East, South-East Asia, Australasia) (Anon, 1952),
which is mainly a pest of Oriental tobacco. When young plants are
attacked, the stems swell to form galls where the larva is feeding and
this causes stunting or death of the plant. The larva penetrates the
inner core of the stem (unlike the superficial burrows of leaf-miner).
Suckers tend to be produced below the gall. In woodier plants the
symptoms of attack are less severe and may not be noticed.
Each female lays 50 to 80 eggs singly on any part of the plant and
these hatch in about a week. The larvae tunnel in leaf tissue and make
their way via a vein and the mid-rib to the main stem, where they
complete their development. The larva prepares an exit hole in the
stem, prior to pupating, to allow the moth to emerge later. The cycle
from egg to adult takes about 1 month.
As infestations usually start in seedbeds and continue in the field,
control measures in both situations are usually applied. Plants
showing symptoms of damage are removed and stalks are destroyed at
final harvest to prevent carry over of the pest to the next planting.
m
Whiteflies
The adult white fly is a fragile insect, 1 mm long and covered with a
finely powdered white waxy bloom. The two main species that feed
on tobacco are Bemisia tabaci (cosmopolitan) (Anon, 1952) and
Trialeurodes vaporariorum (cosmopolitan). The minute eggs are 0.2
mm long, pear-shaped with a stalk, white at first and becoming
brownish later. The first of the four nymphal instars is mobile, having
functional legs and antennae, while the other instars are sedentary
with degenerate legs and antennae. The nymphs are oval in outline
and flattened, giving the appearance of a scale insect. The adult is
formed within the fourth instar nymph and as a result is sometimes
called a 'pupa'. It is 0.7 mm long with red eyes of the adult, obvious
through the translucent pupal exoskeleton.
The sedentary nymphs feed on plant sap and produce honeydew, on
which sooty mold grows and discolors the leaf. They are able to
transmit tobacco leaf curl virus which causes the leaves to pucker,
twist and curl, and in severe infections there may be leafy outgrowths
(enations) from the veins on the underside (see virus diseases in
Chapter 6B of this monograph). If infection occurs when plants are
small, they are severely stunted.
The female makes slits with her ovipositor on the underside of leaves
into which she inserts the stalk of the egg. Each female lays about 100
eggs. The life cycle may be completed in 2 to 3 weeks in hot weather
and up to 6 weeks in colder conditions. Populations build up in warm
and humid conditions; if infested plants are shaken, a cloud of
whiteflies flutters out and rapidly resettles.
Control of whiteflies can be effected by systemic insecticides and by
natural enemies such as the tiny parasitic wasps Encarsia and
Eretmocerus.
Conclusion
As mentioned in the introduction to this chapter, it has not been
possible to include all insect pests found on tobacco in the world.
However, some of the minor pests that have a wide geographical
distribution include the vegetable weevil, mole crickets, white-fringed
beetle, Japanese beetle and seed-harvesting ants. Although not insects,
snails, true slugs (as opposed to the tobacco slug, which is a beetle
larva, see (k) this chapter) and millipedes are minor pests with a wide
distribution.
References
Akehurst, B.C. (1981) Tobacco, 2nd edn. Longman, London and New
York.
Anonymous (1952)Ò Distribution Maps of Insect Pests. International
Institute of Entomology, London, UK (formerly Commonwealth
Institute of Entomology, London). (Looseleaf; maps revised
periodically and new maps added.)
Anonymous (1982) The Locust and Grasshopper Agricultural
Manual. Centre for Overseas Pest Research, London (now Natural
Resources Institute, Chatham Maritime, UK).
Anonymous (1990) Section I: Insect and mite pest control. In: Flue-
Cured Handbook of Recommendations and Burley Handbook of
Recommendations. Tobacco Research Board, Harare, Zimbabwe.
(Looseleaf; updated periodically.)
Anonymous (1993) The use of published guidelines in the selection,
application and handling of agrochemicals in leaf tobacco production.
CORESTA Info. Bull. 19931, 2131.

 Page 240
Anonymous (1995a) Tobacco Field Guide for Central and Southern
Africa. Bayer Zimbabwe (Pvt) Ltd, Harare, Zimbabwe.
Anonymous (1995b) Tobacco disease losses 1993. In: Proc. 36th Tob.
Workers' Conference, Tampa, Florida, January 1995.
Blackman, R.L. (1987) Morphological discrimination of a tobacco-
feeding form from Myzus persicae (Sulzer) (Hemiptera: Aphididae),
and a key to new world Myzus (Nectarosiphon) species. Bull. Ent.
Res., 77, 71330.
Blair, B.W. (1975) Behavioural studies on the larvae of Agrotis
segetum (Denis & Schiff.) and A. ipsilon Hüfn. (Lepidoptera:
Noctuidae): towards better pest management. In: Proc. 1st Congr. Ent.
Soc. Sth Afr. pp. 1923. Stellenbosch, South Africa, SeptemberOctober
1974.
Blair, B.W. (1976) Comparison of the development of Agrotis ipsilon
Hüfn. and A. segetum (Denis & Schiff.) (Lepidoptera: Noctuidae) at
constant temperatures. J. Ent. Soc. Sth Afr., 39, 2717.
Blair, B.W. (1982) Seasonal abundance of Agrotis segetum (Denis &
Schiff.) and A. ipsilon (Hüfn.) in Zimbabwe and a method of
forecasting post-winter population densities. J. Ent. Soc. Sth Afr., 45,
21015.
Blair, B.W. (1986) Strategies to minimize resistance in arthropod
pests to acaricides and synthetic pyrethroid insecticides in Zimbabwe.
In: IVe Congrès sur la Protection de la Santé Humaine et des Cultures
en Milieu Tropical. pp. 2227. Marseille, France, July 1986. Chambre
de Commerce et d'Industrie de Marseille.
Blair, B.W. (1990) Insect and mite pests of tobacco in Zimbabwe:
Description, biology and damage. Tech. Bull. No 1. Tobacco Research
Board, Harare, Zimbabwe.
Blair, B.W. (1994) Legislative control of tobacco pests in Zimbabwe
(abstract). CORESTA Info. Bull., Congr. Issue 36. Harare, Zimbabwe.
Büttiker, W. (1994) Observations on the economic importance and life
cycle of several tenebrionid beetle species in Zimbabwe. Trans. Zimb.
Sci. Assoc., 68, 16.
Chaff, M.S. & Nagarajan, K. (1994) Management of Pests and
Diseases of Tobacco. Central Tobacco Research Institute.
Rajahmundry, India.
Hill, D.S. (1987) Agricultural Insect Pests of the Tropics and Their
Control, 2nd edn. Cambridge University Press, Cambridge, New
York, New Rochelle, Melbourne and Sydney.
Kuitert, L.C. & Tissot, A.N. (1956) Insect pests of flue-cured tobacco
and their control. Univ. Fla. Agric. Exp. Sta. Bull. No 573.
Linker, H.M., Southern, P.S., Melton, T.A. & Smith, W.D. (1989)
Scouting tobacco in North Carolina. Agric. Ext. Serv., N.C. State Univ.
Pub. AG400.
Manley, D.G. (1995) Insect management. In: South Carolina Tobacco
Growers Guide 1995. Coop. Ext. Serv. Pub. EC 569. Clemson
University, SC.
Rabb, R.L., Scott, H.E., Guthrie, F.E. & Smith, C.F. (1959) Tobacco
insects of North Carolina and their natural enemies. Agric. Exp. Sta.
NC Sta. Coll. Bull. No 394.
Reich, R.C. (ed.) (1985) Flue-Cured Tobacco Field Manual and
Burley Tobacco Field Manual. R.J. Reynolds Tobacco Co., Winston-
Salem, NC.
Shaw, M.J.P. (1976) Crop losses caused by insect pests of tobacco.
Rhodesia Agric. J., 73, 141 (abstract).
Shaw, M. (1981) Insecticides money well spent. Zimb. Tob. Today, 4
(7), 9, 15, 17.
Southern, S. (1995) Insect management. In: Flue-Cured Tobacco
Information and Burley Tobacco Information. North Carolina
Cooperative Extension Service, North Carolina State University,
Raleigh, USA (updated each year).
Xuan, T.H. (1989) Tobacco Insect Pests, Diseases, their Natural
Enemies: Identification and Management. National Tobacco
Administration, Quezon City, Philippines.

 Page 241

7B
Stored Tobacco: Insects and Their Control
E.D. Massey
British American Tobacco, R&D
Southampton, UK
Introduction
Two insect species, the tobacco or cigarette beetle (Lasioderma
serricorne) and the tobacco moth (Ephestia elutella) are known to
feed on cured tobacco. Ephestia elutella is indigenous to the northern
hemisphere, being adapted to temperate climes. It is predominantly
reported to feed on leaf stocks and is excluded from the tropics as a
result of its intolerance to high temperatures (Cox & Bell, 1981).
Lasioderma serricorne is particularly suited to tropical regions
(USDA, 1972) and is known to infest and consume all stages of the
product, resulting in spoilage of at least 1% (US&!;300 million) of
stored tobacco stocks per annum (USDA, 1972). Because of its ability
to consume tobacco at all stages of the manufacturing process,
including finished cigarettes, the cigarette beetle may be present in
leaf storages, established in processing facilities, surviving in tobacco
dust accumulations, finished product warehouses and even present in
retail premises and vending machines.
A comprehensive, integrated pest management program is required to
produce a quality product that is free from active insect infestation
and insect corpses or byproducts (excretia and spotting by body oils).
This involves a team approach and must be practised at all stages from
curing on farms, through re-drying, transport, manufacture and
storage of the final product, shipping and wholesale/retail sale. While
chemical treatments such as fumigation and Kabat (methoprene)
treatment are used to produce infestation-free starting material, good
sanitation (housekeeping) is the key to maintaining the subsequent
product infestation free with treatments such as insecticides reserved
for specific circumstances.
Biology of the Cigarette Beetle, Lasioderma Serricorne
a
Morphology and Life History
Lasioderma serricorne (Fabricius), (Coleoptera: Anobiidae) was first
recorded as a tobacco pest in Paris in 1848 (Runner, 1919) and in
America in 1886 (Tenhet & Bare, 1951). The adults are 2.0 to 3.7 mm
long, light to dark brown in color and of ovoid shape, with the first
thoracic segment bent down giving a humped convex appearance (Fig.
7.15). The antennae are characteristically saw-like with equal
segments, in contrast to the club-like segments of the antennae of the
drug store beetle (Stegobium panaceum) which in many other ways
resembles L. serricorne in visual appearance.

Fig. 7.15
Lasioderma serricorne. Taken from Runner, 1919.
Adults fly up to 3 km (Buchelos, 1981), though slowly, during dusk
and darkness whenever the temperature is above 18°C (Howe, 1957).
The females lay between 45 and 116 eggs; each has a waxy shell to
prevent desiccation and is approximately 0.4 mm long by 0.2 mm
wide. Eggs only hatch when the temperature is above 20°C with an
approximate success rate of 50 to 60%, which provides an indication
of why the life cycle is only completed above this critical temperature
(Howe, 1957).
The larvae emerge and consume the egg shells which

 Page 242
have an external flora of yeast-like symbionts that colonize the insect
gut and provide vitamins, hence their ability to survive on
nutritionally poor food sources (Ashworth, 1993a). The larvae
complete four growth stages and attain an eventual size in the region
of 4.5 mm. They are creamy-white to gray-white in color and it is the
larval stage in which the beetle can over-winter in temperate zones
(Ashworth, 1993a). They are negatively phototrophic and seek small
holes, e.g. pack seals to enter items such as packed commodity
(Bovingdon, 1931). It is the larvae that consume tobacco, each eating
13 to 16 mg as they 'wander' in the tobacco mass or outer 50 mm of
tightly packed tobacco strips. They produce the characteristic shotgun
appearance (small holes) of packed leaf and powder debris of tobacco
and excretion products.
The pupal stage of the development occurs within the tobacco or
attached to processing equipment in a cell made of food and excretia
products (Howe, 1957). After 4 to 12 days, the pupae moult to
become adults and emerge mainly during the night with a sex ratio of
1:1 (Sivik, et al., 1957; Samuel, et al., 1984). In general the whole life
cycle is completed within the tobacco, there being little requirement to
leave the product as it will both provide food and harbor potential
mates.
Commencing some 10 hours after emergence, the female releases a
pheromone (4S,6S,7S-serricornin) which attracts the male (Coffelt &
Burkholder, 1972). This chemical, which can be commercially
synthesized, forms the attractant lure of the now standard industry
monitoring tool for this species. Seven days post emergence some
96% of females are reported to have been mated, ensuring the high
success of the species (Levinson & Levinson, 1987).
b
Survival and Development
The life cycle and length of individual stages depend upon
temperature, humidity and nutritional value of the food source
(Ashworth, 1993a). The adults survive between 2 and 7 weeks and
under optimum conditions, 28°32°C and 70% relative humidity, egg
to adult can be completed in approximately 24 days, giving an
increase in the beetle population of 100% per week (Howe, 1957). For
practical purposes, some 11 generations per year at 32°C, 6 at 25°C
and 3 at 21°C have been reported (Howe, 1957).
However, it should be noted that all stages of L. serricorne can
survive between 2°C and 36°C (Fletcher, et al., 1973). At the lower
temperatures (below 18°C), minimal activity will be detected, though
viability will remain which will increase if the ambient temperature is
raised by either seasonal effects or shipment of tobacco stocks from
temperate to warmer climes.
c
Trophism
Its food sources are probably the widest known among insects, but
include most dried and processed plant or animal products (Howe,
1957). For tobaccos which vary in their natural levels of nicotine, they
are reported to prefer high sugar, low nicotine types, being nicotine
tolerant up to 4%, but are unable to survive on a diet containing above
8.25% nicotine (Ashworth, 1993a). However, its food preferences
have never been reported as providing strict protection of tobacco
products against infestation.
Natural enemies of L. serricorne have been reported, the most
common being the wasp Anisopteromalus calandrae which lays eggs
in larvae and pupae (Bare, 1942; Howe, 1957). However, these have
never shown promise for control (Ashworth, 1993a), rather their
presence is often indicative of the presence of L. serricome.
Biology of the Tobacco Moth, Ephestia Elutella
Ephestia elutella (Hübner), (Lepidoptera: Pyralidae), was first
recorded as a tobacco pest in Russia in 1915 and like the beetle feeds
on a range of stored products, being particularly prevalent on cocoa
(Mokrzhetskii & Bragina, 1915; Ashworth, 1993b). Unlike L.
serricorne, E. elutella does not complete its life cycle within the
tobacco, since the adults are fragile and cannot push their way out of
confinement or into packed tobaccos (Ashworth, 1993b). Because of
its preference for temperate zones, E. elutella generally completes one
or possibly two life cycles per year.
The adults are even gray and 10 mm from the head to the tip of the
folded wings. They rest during the day and are most active at dusk and
can live for as long as 3 weeks. Mating takes place 1 to 2 days after
emergence in spring and summer then females lay more than 100
eggs, each approximately 0.5 mm in size, on or near tobacco products
in the first 4 to 5 days of life (Ashworth, 1993b). The eggs hatch after
6 to 7 days and the larvae burrow into the tobacco where they remain
for the next 2 to 3 months, feeding and developing through six stages,
reaching a size of 13 mm. Often the whole leaf is devoured, leaving
only veins and mid-rib (USDA, 1972), with flue-cured and
particularly Oriental tobaccos (high sugars and low nicotine

 Page 243
varieties) appearing to be the most favored food sources (Fraenkel &
Blewitt, 1943). Finished or partly processed products, air-cured or
cigar tobaccos are rarely attacked (USDA, 1972).
Mature larvae leave the product before the onset of winter and enter a
wandering phase, leaving silken threads characteristic of infestation.
They spin a cocoon and diapause (i.e. enter a state of dormancy) for 4
to 9 months on the fabric of the building, packaging material or
racking (Ashworth, 1993b). Pupation of the population is
synchronized by increasing temperature and photoperiod, with a
synchronized population of adults emerging in spring to mate and
complete the life cycle (Ashworth, 1993b).
In a similar way to L. serricorne, females secrete a male long range
attractant and mating behavior-inducing pheromone (Krasnoff, et al.,
1983). This attractive agent has been purified, (Z,E) 9,12-
tetradecadien-1 ol acetate, and used as an attractant lure in monitoring
traps for this insect (Ryan, 1995).
Monitoring
Monitoring for the presence of L. serricorne or E. elutella is a primary
tool for determining both where and when action should be taken to
ensure few or preferably none of these pests are able to damage the
raw material or finished product.
In addition to visual inspections, which should be carried out at each
stage of tobacco processing, pheromone traps are now the principle
tool available as an early warning system for both L. serricorne and E.
elutella, being at least 15 times more sensitive than light traps
(Heeman, 1986).
a
Lasioderma Serricorne
Pheromone traps use the chemical secreted by the female, 4S,6S,7S
serricornin, to attract males which are restrained on an adhesive
surface and can subsequently be counted. These data should be used
to indicate not only the presence or absence of L. serricorne, but
whether the population is increasing or decreasing, reflecting the
potential effectiveness of the control processes in operation.
In comparison to the light traps used to monitor their presence,
pheromone traps have a number of advantages:
They are small and versatile in where they may be placed, e.g. among
processing machinery.
No electricity is required.
They are low maintenance.
Their major disadvantage is that they may be inactivated by dust and
must be replaced if the adhesive capture surface is rendered
inoperative. Monitoring for the adhesive performance is an essential
part of trap maintenance in addition to replacement when the
pheromone is exhausted.
Stocks of pheromone ,traps (adhesive board and lures) should be
stored in the refrigerator prior to use at temperatures between 6 and
10°C (not frozen) and used within 1 year of purchase. In the general
monitoring situation, pheromone traps should be numbered and the
date of placement written on them; they are then placed in a grid
pattern with a spacing of 20 m between traps, or one trap per 3000 m3.
They should be placed among processing machinery and between
tobacco stacks in the warehouse situation, preferably at head height.
However, for the best sensitivity, the traps should be installed on a
backing board with a ledge beneath them on which beetles may first
alight and then crawl into the trap. Placement above heat sources (e.g.
dryers) should be avoided, as should places with large air movements.
Inspection should be on a weekly basis, and the count written both on
the trap and on a plan of the tobacco facility involved. These data
should be under the control of one person per site and should be
regularly reviewed by the local management teams to ensure
coordinated site action. The pheromone lure will eventually become
exhausted and the trap, including adhesive board, should then be
replaced in accordance with the manufacturer's instructions, which
may be after anything from 4 to 8 weeks (Ryan, 1995).
If L. serricorne are detected, then the placement of additional traps to
aid precise location has been found useful. For example, in air
conditioned secondary processing areas, L. serricorne are less likely
to fly to traps in large numbers, but traps included inside a protective
holder placed inside cigarette makers have been found to result in
significant captures, indicating that the product is still at risk, though
the general monitoring program may indicate that the environment is
infestation-free.
While L. serricorne do not fly if the temperature is lower than 18°C in
temperate zones, factories are heated for the comfort of staff and
activity may continue year round, especially in the region of
equipment used to heat tobacco (conditioning and drying cylinders)
(Ryan, 1995). Trap monitoring in these areas should continue year
round.

 Page 244
b
Ephestia Elutella
The discovery and synthesis of female-produced sex pheromone (Z,E)
9,12-tetradecadien-1 ol acetate has again led to the use of pheromone
traps for E. elutella detection (Krasnoff, et al., 1983). Capture devices
can be one-way tunnel systems and a container or adhesive boards, as
used for beetle traps. The pheromone is usually placed on the trap or
incorporated into plastic tubes, rubber septa or impregnated cotton.
Trap spacing, maintenance and recording of captures is the same as
recommended for L. serricorne.
c
Action Thresholds
The level at which action should be taken once captures of L.
serricorne are recorded will depend upon the facility and geographical
location of the facility in question. The goal is to have all areas in
which the product is present infestation-free. With a systematic
program of control, this is achievable in all areas of storage and
processing in temperate zones, as well as production premises in
warmer climes. Once data are available from a pheromone monitoring
program, e.g. in a storage in tropical areas, the optimum time to carry
out a fumigation can be determined before counts begin to increase in
processing facilities. In the latter, a systematic and thorough
inspection and cleaning program can be used to ensure product
integrity is not compromised and has been demonstrated to be clearly
achievable without impairing production efficiency. Indeed, in
facilities where systematic and thorough cleaning programs are in
place, machinery is less likely to wear and downtime is reduced.
Housekeeping (Sanitation)
As the tobacco stocks journey from farm through manufacture and
eventually to the customer, a systematic program of preventive control
and elimination of tobacco pests has to be practised. The prime tool in
achieving this is to deny the pest a suitable harborage, including food
upon which to survive. While supply tobaccos can be rendered
infestation-free by Kabat treatment or fumigation, removal of
processing debris, i.e. cleaning, is the key element to insure
processing buildings and equipment are not able to maintain
beetle/moth populations.
On the farm, carry-over from year to year is a potential risk of
infestation (Koizumi, et al., 1985). Farmer involvement through
extension programs is a prime area for commencing control programs.
All farm areas in which cured tobacco is manipulated should be
cleaned thoroughly of all leaf debris, which should be burned. The
dangers of throwing leaf debris onto dry soil or grass should be
pointed out and subsequently discouraged. Use of pheromone traps
should be introduced at this point as a tool to indicate infestation.
In the leaf processing plants, inspection programs must be in place to
detect infested consignments at receipt, with a program to deal with
badly infested lots, either rejection and removal from the process, or
fumigation, depending upon the degree of infestation. The plant must
be regularly cleaned throughout the season using the same principles
applied to tobacco manufacturing facilities. Waste material must not
be left to accumulate, but removed regularly (daily) and destroyed,
while a pheromone trapping and subsequent program to respond to
any increase in beetle captures is just as important as when used in
other parts of the supply chain of leaf to the consumer.
During shipment, integrity and cleanliness of containers are important
facets. Fumigation of tobacco consignments prior to shipment is often
used as a precaution to ensure freedom from pests. Only sound, clean
containers should be used and they should be inspected to insure they
are free from odor, taint or previous cargos. Vents should be screened
(8 strands/cm) before shipment to prevent ingress of L. serricorne or
E. elutella, if the container is to pass through areas where these may
be indigenous. A pheromone trap may be placed in the container in
the country of origin and inspected on receipt to provide further
information on the infestation status of the shipment.
Once in the warehouse of destination, prior to use in production,
tobaccos should be inspected for infestation and not introduced into
'clean' stores until staff are satisfied the consignment is infestation-
free, following inspection of between 3% and 10% of incoming cases.
Tobaccos can be segregated into high-risk or low-risk groups,
depending upon previous experience and, if necessary, those from
high-risk areas should be fumigated upon receipt rather than risk
introduction of infestation into previously clean warehouses. In
warehouses, a pheromone trapping system and response program
should again be in place to respond to potential infestation, as it may
arise. As on the farms, warehouses should be inspected and all debris
removed on a regular basis.
In production premises, a systematic program should continue to
combat potential infestation and is almost the last chance to insure the
integrity of the product before distribution to the customer. Premises

 Page 245
should be inspected to determine where tobacco debris may
accumulate. While the areas requiring general cleaning are easily
identified, it is the remaining areas that demand most attention.
Furthermore, with the help and encouragement of management, once
the tasks and reason for them are explained, operatives and
maintenance staff often know the equipment and areas of building
fabric that most accumulate debris. These should be systematically
identified in the production facilities and then cleaned. The build-up
rates of subsequent debris can then be determined, and cleaning
schedules and procedures set and documented to insure they are
removed with the frequency necessary to prevent build-up to a level
that will allow beetles to breed. Once this process is in operation,
regular inspections to insure that the cleaning job is carried out must
be built into the program. If the manager becomes disinterested in the
program, so will the staff carrying out the work.
The documentation and recording of these processes are most
important. It is the only way to insure that areas to be cleaned are not
forgotten and that the job is subsequently and properly carried out.
In tandem to the cleaning regime, the provision of a pheromone trap-
based beetle monitoring program will ensure that tools are in place to
detect beetles if infested material is introduced into a factory. If
systematic cleaning has been carried out, debris which beetles are able
to use as a breeding harborage will be minimal, and this insures that
they are not easily able to become a permanent feature of the
production premises.
Compressed air as a cleaning tool is a misnomer. It only serves to
force debris, which may contain beetle eggs, into further inaccessible
areas and insure distribution of infestation to a wide area. Compressed
air has no place in a planned cleaning program to combat infestation.
Vacuum cleaning is the most appropriate tool and, if necessary,
production of end tools to reach difficult areas should be undertaken.
In insuring a clean factory, appropriate design features of both the
fabric and equipment must be considered. Do not create areas around
machines which are difficult to clean. For example, cigarette makers
should be at least 15 cm off the floor to facilitate cleaning underneath.
Work on the principle that if one can easily see debris then there is a
greater chance of it being properly removed with the correct
frequency. Ducting and similar items should be designed to reduce
dust settling on the upper surface and should be subject to a regular
cleaning schedule. Electrical control panels should not be situated up
against walls with a difficult to clean space underneath or behind.
Smooth, easy to clean surfaces should be created and the wall/floor
junction should be curved, not a right angle. This list is by no means
exhaustive, but indicates the principle design features which make
cleaning for infestation control easier (Ryan, 1995).
Treatments to Eliminate Live Infestation
These treatments fall into two main areas:
(1) those that may be used on tobacco, e.g. fumigation, temperature
manipulation and insect growth regulators;
(2) those that may be applied to surfaces and the air space, e.g.
conventional insecticides, but would not be permitted to come into
contact with tobacco or materials intended for the production of the
product or the packaging.
a
Temperature Control
Freezing
A range of temperatures has been used to kill all stages of the cigarette
beetle and the essential element is to ensure that all parts of the
tobacco to be treated reach the critical temperature (Ryan, 1995). To
this end, a thermocouple probe or similar device should be included at
the center of the tobacco package to be treated and attached to a meter
to enable these critical temperatures to be monitored and determined
to have been held for the appropriate time.
To date, the following minimum freezing schedules have been
demonstrated to ensure tobaccos are free from beetle infestation:
-18°C for 5 days
-20°C for 48 hours
-25°C for 18 hours
To these times must be added the time necessary for the center of the
tobacco package to reach these critical temperatures. For example, it
has been demonstrated that one 100 kg case of cut tobacco placed in a
-25°C room requires over 8 days for the core to reach -18°C.
Heat
Heating as part of tobacco processing has been demonstrated to kill all
stages of the life cycle of beetles. However, the exact length of time
the tobacco is at

 Page 246
these temperatures, either in the re-drying plant or during primary
processing, is subject to wide variation and cannot be fully relied upon
to provide an infestation-free product (Ryan, 1995) unless tight
process controls are in place.
b
Conventional Insecticides
In most countries the use of these materials is in highly regulated
processes and they must be applied only by trained certified
personnel. The insecticide label and instruction manual must be
followed at all times (CORESTA, 1993).
Many formulations have been used against L. serricone and E. elutella
(Ryan, 1995), though the choice of active ingredient would now be
considered to be either natural pyrethrums synergized with piperonyl
butoxide or synthetic pyrethroids which penetrate the insect cuticle
and act as nerve poisons to kill the insect (Ryan, 1995). Formulations
available are divided into use as space sprays or surface treatments;
specifically natural pyrethrums are the first choice of agents for space
spraying as they lack any residual activity, while synthetic residual
forms may be used for hard surface treatments (Ryan, 1995).
c
Space Sprays
When required, these are usually best applied in evening periods when
insect flight is most active. They may be successful at reducing the
total population if repeated at an appropriate frequency, but will only
kill those insects flying at the time of application. Specifically, they
will not penetrate harborages or tobacco consignments to kill insects
contained therein; indeed tobacco, finished product or elements to be
used directly with the product should be removed or covered up prior
to space treatments to avoid possible contamination of the product.
The prime use of these formulations is considered to be to contain an
outbreak of infestation while the area is thoroughly cleaned to remove
tobacco deposits that harbor the infestation in production areas. In a
leaf warehouse where bales of infested material are present, they can
be used to prevent the infestation spreading while affected
consignments are fumigated.
Application of formulations in the region of 1:10 for
pyrethrins:piperonyl butoxide have been shown to be effective at rates
of 4 mg/m3 of active ingredient against L. serricorne and 1 mg/m3 for
E. elutella. Ultra low volume (ULV) applications using droplet sizes
of <25 µm are generally found to be most effective, though some
formulations are specifically suited to use with thermal fogging
apparatus (Matthews, 1992).
d
Surface Sprays
As the name implies, these are applied to surfaces in anticipation of
the insect pests crawling over the surface and picking up a lethal dose
of the agent. This requires a residual level of insecticidal activity that
depends upon the active ingredient, the formulation and the nature of
the surface to be treated, including porosity and chemical composition
(Beesley & Chadwick, 1986; Chadwick, 1986).
Usually formulations are applied to just short of runoff, hence taking
into account the porosity of the surface. As with the use of all
pesticides, the manufacturer's instructions for dosages to be applied,
safety precautions and appropriate protective equipment must be
followed to insure responsible use (CORESTA, 1993).
e
Insect Growth Regulators
Methoprene, an insect growth regulator, has the ability to halt the life
cycle of target insects and is particularly effective against both L.
serricorne and E. elutella. The mode of action is to prevent the larval
form developing into adults.
In treated and untreated tobaccos adult L. serricorne may lay eggs;
these hatch and develop into larvae which feed upon the tobacco,
causing degradation of the raw commodity. In untreated tobaccos, the
fourth instar larva will enter a pupal phase and metamorphose in the
pupal cell to an adult and continue the life cycle. In methoprene
treated tobaccos, larvae that have ingested methoprene do not
metamorphose and successive generations do not develop (Long, et
al., 1978). The compound interrupts the pathway and breaks the life
cycle of the insects. Two formulations of methoprene, Kabat Tobacco
Protector (usually applied directly to the tobacco leaf during
processing) and Dianex Residual Spray (used as a space fogging
agent), are used to control L. serricorne and E. elutella.
Methoprene has been registered for use in many countries and is
approved by the World Health Organization for addition to drinking
water at 1 µg/g for the control of mosquitoes (Ryan, 1995).
Methoprene is degraded by sunlight, but the conditions of storage of
tobacco insure that if correct levels are applied during the re-drying
process, 5 to 10 µg/g (ppm), these are still effective 2 years later
(Ryan, 1995).
The key to effective infestation control with Kabat is

 Page 247
uniform application to the tobacco crop during the threshing process,
applied at the end of the drier by a system to first mix the Kabat
formulation (5% w/w with ethyl alcohol) with water and then spray
the mixture evenly onto the tobacco. With its low vapor pressure,
methoprene has been demonstrated not to migrate to any significant
degree within a tobacco bale (Ryan, 1995). Controlled application is
achieved by monitoring the tobacco throughput via a weigh conveyor
linked to the spray pumping system. The tobacco throughput and
spray volume data should always be recorded and archived to indicate
application levels which are then further checked by analysis of the
tobacco samples.
Kabattreated tobacco should always be stored separately from
untreated tobacco. If nonKabat-treated stocks containing beetle are
present in the-same warehouse this will serve to constantly re-infest
the Kabattreated material with cigarette beetles, these will develop up
to the larval stages which will degrade the tobacco and may reach
sufficient levels to cause spotting and staining.
In a factory situation, the Dianex formulation of methoprene has been
used as a space spray and resulted once in an 89% reduction in insect-
related consumer complaints (Hutchens, 1990). The key, however, is
to maintain a 10 µg/g level in the surface dust to insure effective
control. As with most aspects of infestation control, insuring that
adequate data are available to confirm the correct levels are present to
give the control required is paramount.
f
Fumigation
Fumigants are gases which will penetrate to all parts of the tobacco
mass and, when used correctly, will kill 100% of all stages of L.
serricorne and E. elutella. The main fumigant for use with tobacco is
phosphine gas. Phosphine will kill most life forms, including man,
and should, therefore, only be used by trained staff who, in most
countries, require government approved training and subsequent
registration (Anon, 1996). This brief description serves only to
explain the principles of fumigation so that personnel can
communicate better with trained fumigators.
The preparation is supplied as a formulation based upon aluminum or
magnesium phosphide. These formulations, when removed from their
sealed containers, absorb water from the atmosphere which then reacts
to form phosphine gas and leave a residue of aluminum or magnesium
hydroxide. Magnesium phosphide generates phosphine gas faster and
more consistently at lower temperatures than aluminum phosphide.
Aluminum phosphide is not recommended for use when the
temperature is below 15°C and magnesium phosphide is not
recommended below 7°C (Howe, 1974; Sullivan, 1985; Ryan, 1995).
Phosphine gas has a garlic-like smell and will corrode copper, alloys
of copper and precious metals, such as gold and silver; consequently
electrical installations require protection from phosphine gas during
fumigations. For this reason tobacco processing equipment cannot be
fumigated with phosphine.
The preparations come in tablets or a more packaged form, such as
bags and plates. These packaged forms minimize potential
contamination of tobaccos by containing the spent fumigant residues
in the bag or plate and are the preferred form to use.
The correct use of phosphine generating preparations is paramount to
ensure that phosphine-resistant strains of beetles are not selected
(Taylor & Halliday, 1986). There is a CORESTA group contracting
with a lab to examine the potential phosphine resistance issue. The
dose of aluminum or magnesium phosphide preparation to be applied
to a fumigation should be sufficient to generate 1 gram of phosphine
gas (710 µg/g) per cubic meter of the volume to be fumigated, which
also includes the volume of the tobacco in the stack or warehouse
under fumigation.
Fumigations are carried out in a closed space at atmospheric pressure.
The space is sealed to prevent leakage of the phosphine gas to insure
that the fumigation is both effective and safe. In some cases whole
buildings are sealed or alternately fumigations may be carried out
under gas-proof sheeting (150 µm polyethylene) sealed to the ground.
Barriers and warning notices should be placed to insure that all
personnel are excluded from a region where the phosphine gas
exceeds 0.1 µg/g (NIOSH, 1990). Indeed, operatives in fumigations
will wear respiratory protective equipment whenever concentrations
exceed this limit. Full face canister apparatus must be used when the
phosphine gas levels are in the range of 0.1 µg/g to 15 µg/g. Self-
contained breathing apparatus (SCBA) must be used when the
concentration exceeds 15 µg/g (NIOSH, 1990).
It is essential to monitor the concentration of phosphine gas during a
fumigation using gas detection tubes or an electronic meter (Ryan,
1995) to insure that a minimum of 200 µg/g of phosphine gas is
maintained at the center of the tobacco bales for at least 48
consecutive hours within a longer than 4-day fumigation. This insures
100% kill of all the stages of the tobacco beetle (Childs, et al., 1973;
Howe, 1974; Hole, et al.,

 Page 248
1976). Sample gas lines should be included in all fumigations to both
the air space and the center of tobacco bales through which the gas
can be monitored. To insure a successful fumigation these timings and
concentrations of phosphine fumigant to be achieved should not be
altered. For phosphine, Harber's rule (concentration × time of
fumigation = a constant) does not apply; the length of time required
for an effective fumigation cannot be reduced by increasing the
concentration applied, the concentrations and time of the fumigation
given above must be rigidly enforced.
At the end of the fumigation the building/stack is vented to release the
phosphine gas. The doors and vents are opened from the outside or the
gas proof sheeting is drawn back by workers wearing respiratory
protection. Aeration is continued for 2 days or until the airspace is less
than 0.1 µg/g (ppm), whichever period is longest. Spent fumigant
should be collected by personnel wearing appropriate protective
equipment and deactivated as recommended in the manufacturer's
instructions (Ryan, 1995).
Conclusions
In this section, an up-to-date review of basic information and
techniques as background for the development of an integrated pest
management strategy to control infestation in tobacco leaf stocks,
premises and products has been presented. The emphasis is on a
structured control procedure at all stages involving the use of cured
tobacco, i.e. from the farm to the finished product. Sanitation is the
major tool for effective control, with the use of pesticides reserved for
specific purposes.
A more comprehensive text covering these topics has been published
and should be consulted for more detailed information (Ryan, 1995).
References
Anonymous (1996) UK Health and Safety Executive, Fumigation
using Phosphine. Guidance Note CS22, ISBN 0-7176-1218-X.
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Ashworth, J.R. (1993b) The biology of Ephestia elutella. J. Stored
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Buchelos, C.T. (1981) Coleoptera populations at flour mills and
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Childs, D.P., Overby, J.E., Cox, E.L. & Niggenegger, D. (1973)
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Cox, P.D. & Bell, C.H. (1981) A Review of the Biology of Moth Pests
on Stored Products. ADAS publication, Slough Laboratory, Berks.
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Fraenkel, G. & Blewitt, M. (1943) The natural foods and food
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Heeman, V. (1986) Insect control: natural chemical attack beetles.
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Hole, B.O., Bell, C.H., Mills, K.A. & Goodship, G. (1976) The
toxicity of phosphine to all development stages of thirteen species of
stored product beetles. J. Stored Prod. Res., 12, 23544.
Howe, R.W. (1957) A laboratory study of the cigarette beetle,
Lasioderma serricorne (F.) (Col., Anobiidae) with a critical review of
the literature on its biology. Bull. Ent. Res., 48, 956.
Howe, R.W. (1974) Problems in the laboratory investigation of the
toxicity of phosphine to stored product insects. J. Stored Prod. Res.,
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Hutchens, H.M. (1990) Advantages of using insect growth regulators.
Paper presented at Federation of Asian and Oceania Pest Managers
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Koizumi, S., Kikukawa, H. & Shimojic, C. (1985) Occurrence and
ecology of pests attacking stored tobacco in growing districts in
Kyushu. Kagoshima Tob. Exp. Stm. Bull., 26, 7586.
Krasnoff, S.B., Vick, K.W. & Mankin, R.W. (1983) Female calling
behavior in Ephestia elutella and E. figulilella (Lepidoptera:
Pyralidae). Florida Ent. 66, 24954.
Levinson, H.Z. & Levinson, A.R. (1987) Pheromone biology of the
tobacco beetle (Lasioderma serricorne F., Anobiidae) with notes on
the pheromone antagonism between 4S,6S,7S-serricornin and
4S,6S,7R-serricornin. J. Apl. Ent., 103, 21740.
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7C
Pesticide Regulations and Their Impact on Crop Protection Strategies
(Minimization of Pesticide Residues)
L. Mueller
R.J. Reynolds Tobacco GmbH
Cologne, Germany

M.R. Ward
Advanced Technologies (Cambridge) Limited
Cambridge, United Kingdom
The Need for Pesticide Regulations
Tobacco production, like the production of nearly all other crops, is
continuously affected by harmful organisms. Therefore, it is generally
necessary to resort to appropriate measures to protect the growing
plant, to secure desired quality and yield levels and to preserve the
crop after harvesting. The use of plant protection agents (pesticides) is
one of the most important ways to achieve this objective.
Applying pesticides appropriately and effectively for the purpose
intended is by no means unproblematic. Because of the potential for
nonbeneficial effects upon the crop itself, on other plants and on the
environment in general (including humans, and specifically people
working with these pesticides), a large number of recommendations,
rules and official regulations are in existence. Today, the use of
pesticides is increasingly seen as an essential part of modern
agricultural systems, such as Good Agricultural Practice (GAP) and
Integrated Pest Management (IPM).
Regulations cover nearly all aspects of pesticide use, though a large
degree of diversity can be observed when comparing different
countries or international bodies. The registration of (new) plant
protection agents is regulated in many countries often in a very
detailed way. The requirements for labeling, packaging and
application procedures are generally defined in the course of the
registration process. Maximum residue levels (MRLs) for pesticides
have been established in several countries for various crops including
tobacco and/or for products made from tobacco. Proper setting of
MRLs ought to be based on the scientific assessment of pesticide
characteristics and in consideration of good agricultural practice. For
sensible enforcement of MRLs, reliable and validated methods should
be available for use in analytical laboratories.
Registration of Pesticides
Most countries require that a pesticide be formally approved before it
can be used by growers and others for the purpose of protecting a
crop. However, procedures vary greatly among various countries.
They may be statutory or voluntary in nature; they may require
specific and detailed, or rather general supporting data and approval
may be granted for specified crops only or for general use. Therefore,
international bodies such as the World Health Organization (WHO),
the Food and Agriculture Organization (FAO) and the European
Union (EU) have for some time been working towards a more
uniform approach.
The first international guidelines, the International Code of Conduct
on the Distribution and Use of Pesticides, were published by FAO
(1985). The Code, endorsed by all the major agrochemical
manufacturers through the Groupement International des Associations
Nationales de Fabricants de Produits Agrochimiques (GIFAP), was an
important step forward in a harmonized system for registration of
pesticides. GIFAP is now known as the Global Crop Protection
Federation (GCPF). The Code addressed the concerns on the


 Page 251
supply of pesticides into countries which had no infrastructure to deal
with registration. It laid down voluntary standards to cover the
manufacturing, distribution, marketing and use of pesticides and relied
on the collaboration between exporting and importing countries
(governments, manufacturers, distributors and users). The Code was
amended in 1989 (FAO, 1990) to include the article on prior informed
consent (PIC), which set down an important principle that no
government should allow the export of a banned or severely restricted
pesticide to a country without first informing the importing country of
these restrictions and receiving confirmation back before proceeding
with the export of the product. This allows countries without testing
and evaluation infrastructures to evaluate the potential risks. This has
been a contributory factor to the worldwide ban of some of the
persistent organic chemicals, especially the organochlorine pesticides,
such as aldrin, dieldrin, endrin and DDT. The current USA list of
banned pesticides has 47 entrants, and this list forms part of the USA
participation in the voluntary prior informed consent procedure. In the
European Union, there are currently 39 banned or severely restricted
dangerous chemicals (Council of the European Communities, 1996),
of which 11 are pesticides covered by the PIC procedure.
Through the Code, governments were encouraged to establish their
own legislation and testing facilities, and the FAO collaborates with
156 member governments on the implementation of the Code and the
incorporation of it into national legislation.
With the increasing concerns for the environment and the potential
adverse effects of some pesticides, both the United States and the
European Union have introduced recent legislation on the registration
of new and the reregistration of existing pesticides. The registration
process has become an expensive and time consuming exercise, and
agrochemical companies are becoming more prudent in their choice of
label approvals. In the USA, all active ingredients of pesticides
initially registered before 1984 have to be reregistered under the
Federal Insecticide, Fungicide and Rodenticide Act (US
Environmental Protection Agency, 1988). This more stringent re-
assessment of pesticide safety is considered to be critical for
protecting human health and the environment. The requirements for
registration and reregistration were further reinforced by the Food
Quality Protection Act (US Environmental Protection Agency, 1996a)
to provide increased protection for infants and children, especially
with regard to residues in food. The new standard introduces the
concept of 'reasonable certainty of no harm' for aggregate exposure
using dietary and other reliable exposure information.
In 1988, approximately 600 groups of related pesticide active
ingredients required re-evaluation. Of those, 200 cases have been
cancelled because manufacturers failed to support them, and of the
remaining 383 cases supported, eligibility decisions have been made
on only 129 cases (US Environmental Protection Agency, 1996b). The
reregistration process requires a full data submission for each
chemical and an examination and risk assessment of any related health
and environment issues. When the EPA has completed this assessment
and is satisfied that the pesticide will not pose unreasonable risks to
human health or the environment, their conclusions are presented as a
reregistration eligibility decision (RED) and the pesticide can then go
to full registration. A number of pesticides previously registered in the
USA for use on tobacco have not been reregistered (monocrotophos,
fensulfothion, azinphos-methyl, malathion and diazinon). In the
future, the use of other pesticides may be withdrawn as a result of the
reregistration program, for example aldicarb, carbofuran, endosulfan
and methomyl (Sheets, 1991).
In the European Union, the Council Directive of 15 July 1991 and its
multiple amendments (Council of the European Communities, 1991)
provide for a community-wide system for controlling the supply and
marketing of plant protection products.
The basic goal of the Directive is the establishment of a European
Union list of authorized active substances (the so-called Annex I). For
this purpose, a Community procedure is laid down for assessing
whether an active substance can be entered on the Community list.
The information is detailed which an applicant must submit for
admission of the substance to the list (to date, summer 1998, only two
chemicals have been entered into Annex I). Similarly, the Directive
defines the procedure and the data requirements for the authorization
of a plant protection product, i.e. the formulation of an active
ingredient found on the Community list. The Directive also calls for
periodical review, in the future, of the substances on the list and
provides for an emergency authorization mechanism in cases of
unforeseen dangers to plant production.
The compilation of necessary tests, analyses and data for ingredient or
formulation approval is quite impressive indeed and laid out
meticulously in the Directives. In summary, it is required to
demonstrate, on the basis of the information submitted, that the
ingredient or formulation, when used as prescribed and

 Page 252
with application of the principles of good plant protection practice as
well as, whenever possible, of integrated pest control:
is sufficiently effective;
has no unacceptable effect on plants or plant products;
does not cause unnecessary suffering and pain to vertebrates to be
controlled;
has no harmful effect on human or animal health, directly or indirectly
(e.g. through drinking water, food or feed) or on groundwater;
has no unacceptable influence on the environment, having particular
regard to the following considerations: its fate and distribution in the
environment, particularly contamination of water including drinking
water and groundwater, and its impact on nontarget species.
Among the many other aspects addressed by the Directive, three are
particularly important for the following discussion. Where relevant, an
acceptable daily intake (ADI) for man must be determined, MRLs in
the agricultural products referred to in the authorization must have
been provisionally established and appropriate methods in general use
must be available to determine residues, which are of toxicological or
environmental importance, resulting from authorized uses.
Workers' Safety
An important part of any legislative framework is the protection it
affords to those working with or who may come into contact with
pesticides. The basis for some of this protection and safety is in the
registration procedure for individual products and the information that
is required to go on the product label. In many countries where the
principles of the FAO Code of Conduct have been adopted, the label
constitutes a legal document, and they generally have the same format
and information on them. The identified hazards to humans, animals
and the environment have to be stated on the product label along with
brief emergency first aid details. The precautionary statements are of
two kinds: mandatory statements that include the word must; and
recommendations that include the word should. There is often a re-
entry statement which specifies how much time must pass before a
pesticidetreated area is safe for entry by a person not wearing
protective clothing. In addition, when there is a possibility that the
crop may be fed to animals or eaten or handled by humans, there is a
harvest interval statement giving the period of time that must pass,
after treatment, before the crop is harvested. This information on the
label provides the baseline of safety for the use of the pesticide. The
information is supplied by the manufacturers, endorsed by the
registration procedure and places a responsibility on the handler or
user of the pesticide to understand and follow the instructions.
Further legislation for greater protection of people using or who may
come into contact with pesticides is in place in many countries of the
world under different guises of health and safety legislation. The
procedures to be followed are often based on the toxicity of the
chemical in question, be it an agricultural or industrial chemical.
There are two main classification systems for categorizing the toxicity
of a pesticide, and this in turn dictates to a great extent how the
chemical should be handled at the different stages from
manufacturing, transportation, storage and sale to mixing and
application.
The World Health Organization received requests from member
countries of the United Nations to develop a pesticide classification
that would distinguish between the more hazardous and the less
hazardous pesticides (WHO, 1973). A scheme was developed based
on the LD50 or acute toxicity in rats (mg/kg body weight) following
oral or dermal exposure to liquid or solid pesticides (Table 7.1). A
similar approach has been adopted by the EU (Council of the
European Communities, 1978) with three main categories, harmful,
toxic and very toxic, with additional information on other hazards
(highly flammable, corrosive, irritant, explosive and oxidizing). The
hazards have to be represented pictorially on all registered pesticides.
The United States Environmental Protection Agency has four classes,
based not only on acute toxicity through the oral, dermal and
inhalation routes but also considering eye and skin effects (Table 7.2).
The personal protective equipment (PPE), required to be worn by all
operators using pesticides under the EPA Workers Protection
Regulations, is based on the classification shown in Table 7.3.
In the USA, the Workers Protection Act (US Environmental
Protection Agency, 1994) lays down specific training requirements for
workers who, as part of their employment, are either handlers of
agricultural pesticides (and are likely to come into contact directly
with the pesticide) or are working on a farm where pesticides are
used. The Act attempts to reduce the risk

 Page 253
Table 7.1 World Health Organization classification of pesticides.
Class LD50 for the rat (mg/kg bw)
Oral Oral Dermal Dermal
solids liquids solids liquids
Ia Extremely hazardous £5 £20 £10 £40
lb Highly hazardous >5£50 >20£200 >10£100 >40£400
II Moderately hazardous >50£500>200£2000>100£1000>400£4000
IIISlightly hazardous >500 >2000 >1000 >4000
Product unlikely to present acute
hazard in normal use

Table 7.2 United States Environmental Protection Agency toxicity

classification.
Hazard indicator Toxicity categories
I II III IV
Oral LD50 (mg/kg) <50 50<500 500<5000 ³5000
Dermal LD50 <200 200<2000 2000<20 000 ³20 000
(mg/kg)
Inhalation LD50 <0.2 0.2<2 2<20 ³20
(mg/kg)
Skin effects (at 72 corrosive severe moderate slight
hours) irritation irritation irritation
Eye effects*
irritation: corrosive persisting reversible none
corneal opacity: not reversible reversible none none
'Signal word' DANGER/POISONWARNINGCAUTION CAUTION
*Note: Reversibility of corneal opacity and persistency of irritation are both
measured over a period of 7 days after contamination.

Table 7.3 Personal protection equipment requirements.

Acute toxicity category
Toxicity I II III IV
Dermal/skin Coverall, long Coverall, short Long sleeves, Long sleeves,
irritation sleeves, long pants sleeves, short pants long pants long pants
 Chemical-resistant Chemical-resistant Shoes Shoes
footwear footwear
Chemical-resistant Chemical-resistant Chemical- No
gloves gloves resistant requirement
gloves
Inhalation Respiratory Respiratory No No
toxicity protection protection requirement requirement
Eye Protective eye Protective eye wear No No
irritation wear requirement requirement

of exposure to handlers of pesticides by stipulating the requirements

for PPE and requires notification procedures to be adopted for all
spraying operations and the re-entry times into sprayed areas.
In Europe, there is no direct legislation safeguarding pesticide
handlers, but Directive 89/656/EEC on the use of personal protective
equipment (PPE) at the work place (Council of the European
Communities, 1989) has been incorporated into the national law of
member states and covers the use of all dangerous substances. In other
countries of the world, similar legislation exists which requires the
appropriate training and/or certification for all pesticide handlers.

 Page 254
Industry Stewardship
Although these and many other pesticide registration and re-
registration schemes broadly follow the FAO Code of Conduct, there
still remain wide discrepancies among schemes, and harmonization is
still some way from being realized (Anon., 1992). Furthermore, in the
less developed countries legislation on pesticide control is still in its
infancy, and over 40% of these countries are without official approval
schemes, due to lack of either support or resources (Ledru, et al.,
1994). The FAO has conducted a review on schemes existing in a
number of countries, selected on the basis of their geographical spread
and technical experience (FAO, 1995), and the Organization for
Economic Cooperation and Development (OECD) has reviewed the
data requirements for registration for different countries (OECD,
1996). This variable response on legislation for the control and use of
pesticides highlights the necessity for appropriate stewardship from all
companies and trade organizations either directly involved in
pesticides or involved in the international trade or production of plant
products.
To this end, the agrochemical industry's response to the FAO Code of
Conduct was the launch, by GIFAP, of the Safe Use Initiatives. These
were initiatives based on the provision of training programs for local
farmers with involvement of national governments, NGOs, industry
and aid donors. The objectives of these programs were:
(1) the protection of man and his environment;
(2) prevention and treatment of chemical accidents;
(3) recycling and disposal of empty containers;
(4) enforcement of regulatory legislation;
(5) education and training of farmers, retailers, doctors and
schoolchildren;
(6) creation of poison centers.
These initiatives have had notable success in Thailand, Kenya and
Guatemala, where several hundred thousand farmers have been
trained, and further programs in these and other countries are planned
for the future. Specialized courses and systems of accreditation are
introduced for stockists and retailers of agrochemicals as well as for
doctors and paramedics on the diagnosis and treatment of crop
protection products poisoning.
The tobacco industry, particularly through CORESTA (a worldwide
organization which supports international cooperation in research on
tobacco), has been proactive in encouraging the safe use of pesticides.
In 1993, CORESTA published and widely distributed 'The use of
published guidelines in the selection, application and handling of
agrochemicals in leaf tobacco production' (CORESTA, 1993) (at
present available in English, French and Spanish) to increase the
awareness of all sections of the tobacco industry.
Acceptable Daily Intake, Maximum Residue Level and Theoretical
Maximum Daily Intake
It is prudent to assume that the use of plant protection agents generally
leaves residues on the crop of the active ingredient(s) and/or of
degradation or conversion products. Minimization of residues on
crops and in finished products is one of the most important objectives
of responsible use of pesticides. The degree to which such residues
can be expected depends on a number of factors.
First of all, residue levels depend upon the chemical and biological
properties of the molecule itself. Many organophosphate insecticides,
for instance, are very labile compounds so that residue build-up is
rarely observed. The opposite is true for numerous organochlorine
compounds; the persistence of substances like DDT is notorious (and
has led to their ban for almost all purposes).
The right philosophy of pesticide application, as expressed in the
practical principles of GAP, is another important factor.
'Good Agricultural Practice in the use of pesticides includes the
nationally authorized safe uses of pesticides under actual conditions
necessary for effective and reliable pest control. It encompasses a
range of levels of pesticide applications up to the highest authorized
use, applied in a manner which leaves a residue which is the smallest
amount practicable.' (FAO/WHO, 1995)
In particular, the strict observation of dose recommendations and
application-harvest intervals contributes significantly to the
minimization of residue formation. Climatic and weather conditions,
in addition, may have considerable impact.
Residue levels are influenced by the circumstances of crop storage
and processing. Under certain conditions, residues may become
significantly reduced in finished products compared to original
harvested material. This aspect is particularly important for tobacco
and tobacco products.
In any case, it is mandatory for consumer safety to understand and
control any possible adverse health effects from pesticide residues.
Three characteristic

 Page 255
numbers have emerged as the most important criteria for the
assessment and management of risk from pesticide residues.
The ADI is the amount of an active ingredient which can be consumed
daily by an individual during his/her lifetime with the practical
certainty that harm will not result from ingestion. The ADI is
generally given in milligrams per kilogram of the consumer's body
weight. It is based on the experimental determination of the no
observed effect level (NOEL) in the most sensitive animal species
investigated or, in rare cases, on direct observations in man. The
NOEL, in turn, is defined as the highest amount of a chemical which,
in chronic toxicological feeding, drinking or inhalation studies, causes
no significant adverse effects on morphology, development, growth,
life span, biochemical parameters or functional capacity of individuals
of the target species.
To allow an appropriate safety margin for extrapolation from the
animal species to man, a factor of 10 is routinely applied. This step is
taken once more to take into account interindividual sensitivities in
man. Consequently, in many cases the ADI has been set at 100 times
the NOEL, which was found for instance in a study with a rodent
species. The ADI is a characteristic of a given chemical and, in theory,
requires reevaluation whenever new meaningful experimental data
become available.
The second characteristic number, the MRL, originates from specific
agricultural studies using the pesticide in question. Today, a standard
approach is generally pursued in the development of (new) pesticides.
First, field trials under strict observation of GAP demonstrate which
residue levels are found on crops of interest under a variety of realistic
production conditions. To that extent, national and international
guidelines are in existence which define specifics such as the required
number of individual trials, the desirable geographical or climatic
spread across a country or a region and the duration of specific pre-
harvest intervals. These trials demonstrate which residue levels are
realistically found on crops for the pesticide under examination and,
taken together, allow the investigators to calculate an empirical MRL
for the pesticide on a crop.
Using the determined MRL value and considering human
consumption patterns for an agricultural product (or for finished
goods made from it), a theoretical maximum daily intake (TMDI) is
now estimated. This estimate, if done scientifically, takes into
consideration factors such as the frequency of ingestion of a crop or
finished product, the amounts typically consumed, the handling of the
product before consumption (e.g. washing or heating) and specifics of
the uptake (transfer) of a residue to the human body. The latter aspect
is particularly relevant, though complex and frequently poorly
understood, if a product is consumed through the process of smoking.
A crucial step in the registration process is the comparison of the
TMDI value to the ADI level. If the TMDI value does not exceed the
ADI level, consumer safety requirements are generally considered to
be fulfilled. In cases where the TMDI value is higher than the ADI
level, refined calculations (looking at more specific situations) or new
trials under more restricted conditions of pesticide use (e.g. lowered
dose regimens or longer pre-harvest intervals) may lead to a new set
of data for the safety assessment.
For the practical purpose of consumer protection, pesticide residues
are defined, in a relatively small number of countries, in terms of
regulatory MRLs. These are the highest concentrations of a pesticide
residue, expressed in milligram per kilogram (ppm), recommended
(by agreement of responsible parties) or permitted (by law) in or on a
crop or finished product. Ideally, these regulatory MRLs are based on
the findings from studies of the pesticide on various crops conducted
under strict observation of GAP. Generally and logically, different
MRLs exist for the same pesticide on various crops, and their
numerical values can, and often do, vary considerably.
Looking at existing regulations with MRLs for tobacco or tobacco
products, it is obvious that these MRLs are not all based on proper
scientific or technical data. Particularly for pesticides which have been
in use for some time, the absence of useful data from agricultural
trials, and/or the lack of analytical methods for the determination of
residues, constitute major obstacles. In these cases, regulatory MRLs
are often set either on the basis of traditional numbers found in other
countries, or with the intent to keep MRLs at the lowest imaginable
level (e.g. 0.01 ppm). As a result of this, regulatory MRLs for the
same pesticide on tobacco (products) often vary widely amongst
different national regulations, and are in several instances simply not
practical, even for tobacco grown under the conditions of strictly
observed GAP.
International Scientific Bodies Involved in the Regulation of Pesticide
Residues
Today, the regulatory playing field for foods and other products
intended for human consumption has

 Page 256
numerous international and national players, including several well
established scientific bodies. Increasingly, a global or cross-national
approach dominates the activities and pushes the development of
philosophies, methods and regulations to new frontiers.
Two large: organizations provide momentum and structural support
for these activities: the World Health Organization (WHO) and the
Food and Agriculture Organization of the United Nations (FAO).
An interesting example of their co-operation is the Joint FAO/WHO
Meeting on Pesticide Residues (JMPR). Dedicated to the promotion of
the rational and safe use of pesticides, its experts develop evaluations
and recommendations for MRLs in foods. For this, JMPR relies on
two scientific expert groups: an FAO Expert Panel collects and
interprets (residue) data from field trials of pesticides on specific
commodities, conducted under different climatic and environmental
conditions, following the rules of GAP, and a WHO Expert Group is
responsible for the setting of ADIs based on data from toxicological
studies with animals and epidemiological observations in humans.
JMPR is assisted by invited experts and makes use of all authoritative
sources of scientific information. Doubtless, it is the area of risk
assessment where the main expertise and responsibility of JMPR
resides.
Conclusions and recommendations which result from the annual
meetings of JMPR, held since 1961, are published by the co-
sponsoring organizations, WHO and FAO, in the form of reports and
monographs. The primary users of JMPR's output are WHO itself, the
member countries of FAO and the Codex Alimentarius Commission
(specifically its Committee on Pesticide Residues).
Another activity which is sponsored by WHO, jointly with the United
Nations Environmental Program (UNEP) and the International Labor
Organization (ILO), is the International Program on Chemical Safety
(IPCS). Data collection and risk evaluation for chemicals of
importance; prevention, management and treatment of chemical
emergencies; and the development of manpower and technical
methods are the major tasks of this program. Following a conference
in 1992, the IPCS increased emphasis on pesticides by creating the
Joint Meeting on Pesticides (JMP). This consists of the IPCS Core
Assessment Group, which meets as two sub-groups working on
toxicological and environmental assessments (the first meeting was
held in 1994). Additional support is provided by panels on public and
occupational health as well as on environmental issues. Naturally, a
close liaison exists between the JMP and the JMPR.
UNEP also sponsors the International Register of Potentially Toxic
Chemicals (IRPTC), established in 1976. This program collects data
on all kinds of chemicals in a form readily accessible to interested
parties and identifies gaps in the existing information. Furthermore, it
pursues educational goals regarding the risk, control and regulation of
hazardous chemicals. In its activities, IRPTC also addresses issues
related to pesticide use.
A very active scientific organization under the auspices of WHO is the
International Agency for Research on Cancer (IARC), which was
founded in 1965 and is located in Lyon, France. This agency is
dedicated to the evaluation of carcinogenic risks of chemicals for
humans and is known for the production of useful monographs on its
various research topics. Although not systematically dealing with
pesticides, IARC has issued several noteworthy publications in this
field, such as the one published in 1991 on 'Occupational exposures in
insecticide application, and some pesticides'.
A United Nations food standards program set up in 1963, the Codex
Alimentarius Commission (CAC), is about to gain more and more
influence over the way countries establish their standards for food
safety and quality. It is also jointly run by the WHO and the FAO.
About 30 committees subsidiary to the Commission deal with general
subjects, specific commodities and regional problems. Committee
membership is from more than 130 countries and includes
representatives from government, public interest groups, industry and
academia.
The CAC sets standards which are only recommendations. There is no
obligation for governments to transform Codex standards into national
regulations. In fact, Codex standards may be quite different from
existing national standards, which are sometimes much higher or
considerably lower. However, with the new approaches for free world
trade, as heralded by GATT (now WTO), harmonization of national
regulations becomes inevitable, and the Codex standards are likely to
become more normative in nature.
A committee of special interest is the Codex Committee on Pesticide
Residues (CCPR). Created in 1966, the committee is charged with the
preparation of priority lists of pesticides for evaluation by JMPR, with
the establishment of MRLs in foods for human and animal nutrition,
with the consideration of methods of sampling and residue analysis,
and with the establishment of safe limits for environmental and
industrial contaminants showing similarity to pesticides. In
performing its tasks, CCPR follows the recommendations

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of JMPR. Clearly, the role of CCPR is in the management of risk from
pesticides.
All the scientific bodies presented above deal with pesticides, and
several of them particularly with pesticide residues. None of them,
however, specifically deal with pesticide residues on tobacco
(products). It is particularly important to recognize that tobacco is not
within the scope of Codex Alimentarius. Nevertheless, their activities
are of great importance for the tobacco industry, particularly the work
of JMPR and the CCPR. Their critical appraisals (especially those
with an outcome raising concern) have an impact on all applications
of a given pesticide or group of chemically-related pesticides.
Indications of increased risks from certain levels of pesticide residues
are relevant for all agricultural uses, and the ADIs discussed and set
by these bodies, in practice, are immediately applicable to judging
existing MRLs on tobacco (products).
It is therefore in the interest of responsible tobacco cultivation that
especially JMPR and CCPR are provided with the best possible and
most up-to-date scientific and technical information on the chemistry
and toxicology of important tobacco pesticides, on the mechanism of
action, the mode of application and the metabolic fate of pesticides
which are significant for tobacco growers, even if their use on tobacco
is not specifically considered and evaluated by these organizations.
An example of the role of JMPR in the assessment of an important
group of fungicides, the dithiocarbamates, and their metabolites is
related in a publication by Vettorazzi, et al. (1995). Dithiocarbamates
had come under severe attack due to an experimentally observed
carcinogenic effect on the thyroid gland of rats. It was only after
careful review of the scientific evidence by JMPR at its 1993 session
that the toxicology of dithiocarbamates was assessed with an outcome
allowing their continued use, including their application in tobacco
production.
Existing Regulations for Maximum Residue Levels of Pesticides on
Tobacco and Tobacco Products
There are several countries with existing regulations which
specifically apply to tobacco and/or tobacco products. All of them
have rather straightforward and legally binding ordinances or decrees
in place. Germany, for instance, has implemented an approach which
combines a legal regulation with a voluntary agreement. The
Ordinance on MRLs in § 5 (1) and Attachment 7 specifies permitted
MRLs for 14 individual pesticides or groups of chemically-related
pesticides (the list began with 11 tolerances in 1978) (German
Verordnung, 1994). Many of them are traditional organochlorine
compounds (like aldrin, dieldrin, chlordane, DDT,
hexachlorocyclohexane isomers, heptachlor and polychloroterpenes).
Others, such as aldicarb and the more recently added terbuphos,
belong to other chemical classes of compounds. It is important to
understand that the existence of MRLs for the pesticides in question
does not indicate that the pesticides are all approved for use on
tobacco. On the contrary, substances like DDT and dimefox have now
been banned for a number of years. In any case, it is punishable to
exceed stated MRLs because they are laid down in an Ordinance.
With regard to pesticides (other than those in Attachment 7) which are
not approved in Germany for use on tobacco but may be found in
tobacco products (particularly those made from imported tobacco), § 5
(2) of the Ordinance points out that commercial marketing is
acceptable as long as the amounts of residues found on and in tobacco
products are not likely to pose a risk to (human) health.
Based on this provision and as a supplement to the legal regulation
discussed above, the German cigarette industry and the manufacturers
of plant protection agents, in co-operation with the German Federal
Health Office, agreed, in 1978, on recommended MRLs for 64
pesticides (Wittekindt, 1986). Examples of agents of particular
importance which are controlled in this way are carbaryl, the
dithiocarbamates, endosulfan and maleic hydrazide (with an 80 ppm
MRL which has subsequently served as an example for regulations in
several other countries). The agreement was put in place with special
attention to the use of pesticides by tobacco growers in countries other
than Germany and the controlling of pesticide residues on imported
tobacco. Consideration of these tobacco growers' justified needs and
of appropriate agricultural practices played an important role in
setting these MRLs. Although not legally binding, and therefore
without the threat of punishment, experience in Germany over many
years has shown that the tobacco industry complies with the voluntary
agreement as strictly as with the Ordinance.
It is most important to recognize that the MRLs in Germany, whether
permitted or recommended, do not apply to the crop (tobacco in
whatever state of harvesting, curing or storing) but to finished tobacco
products. Specifically, the MRLs relate to the tobacco

 Page 258
content in the product and are expressed as milligram per kilogram (or
ppm).
In Switzerland, MRLs were stated for two crop protection agents in
the Ordinance on Residues and Ingredients in Foodstuffs of 1986
(Swiss Verordnung, 1986). These were a 50 ppm MRL for
dithiocarbamates in tobacco products and a 0.1 ppm MRL of fatty
alcohols (described as octanol/decanol) on tobacco leaf.
Following a revision of foodstuff regulations and the emergence of a
separate Ordinance on Tobacco and Tobacco Products in 1995 (Swiss
Tabakverordnung, 1995), no MRLs for tobacco (products) are found
any longer in the latest Ordinance on Residues and Ingredients in
Foodstuffs of 1995 (Swiss Verordnung, 1995). Therefore, it appears
that, at present, no formal MRLs exist in Switzerland for pesticides on
tobacco or tobacco products. Some kind of regulation is expected to
be developed in the future.
In compliance with Directives of the European Union (although
tobacco was not included as a product of plant origin in the Annex of
90/642/EEC), Spain issued the Royal Decree 280 of 18 February
1994, on the establishment of MRLs on vegetable crops (Spanish Real
Decreto, 1994). Tobacco, defined as 'dry product', was exhaustively
considered in the Decree, and MRLs on tobacco are stated (in Annex
II) for not less than 397 agrochemicals. While the broad majority of
these pesticides had been assigned MRLs in the range of 0.01 to 0.1
ppm, there were 18 pesticides with special importance for tobacco
growing, the MRLs of which ranged from 0.5 to 80 ppm (the latter
level not surprisingly assigned to maleic hydrazide).
Residues of pesticides without a listed MRL and not approved for use
in Spain must not exceed the limit of detection of the most sensitive
analytical method available. New pesticides for which an empirical
MRL, based on their use under GAP conditions, has been recognized
in the course of the approval process, must comply with this MRL
until they are formally included in the list (Annex II).
In 1996, the Royal Decree 280/1994 was amended twice (Spanish
Orden, 1996). MRLs on tobacco were modified for seven of the
agrochemicals already included in Annex II of Decree 280/1994. With
regard to tobacco, the only change of importance was the increase of
the MRL for imidacloprid to 5.0 ppm. In addition, 11 new
agrochemicals were included in Annex II. All MRLs for these on
tobacco are between 0.01 ppm and 0.05 ppm. By footnote, the
amended pages of Annex II indicate the regulatory philosophy that
MRLs below 0.1 ppm are considered the limit of analytical
determination.
Also in response to European Union Directives, the regulation of
pesticide residues on foodstuffs was completely renovated in Italy in
1990. The Ministerial Ordinance on Maximum Levels of Pesticide
Residues Acceptable on Products for (Human) Nutrition of July 1990
(Italian Ordinanza Ministeriale, 1990) included over 330 pesticides, of
which 75 carried MRLs specific for tobacco. The regulation was
extended and made more precise in seven subsequent Ordinances and
Decrees (Italian Ordinanza, 1991), bringing the number of all
pesticides considered up to over 370 and those with MRLs for tobacco
up to 93.
It is worthwhile to examine certain details in the development of
pesticide MRLs for tobacco in Italy. The 1990 Ordinance, in
describing the product in question, generally used the term 'tobacco'.
In six cases, however, the term 'tobacco (dried commercial)' and in
one case (maleic hydrazide) the term 'tobacco (finished product)' was
used. An important step of clarification was accomplished in the
Ordinance of 5 August 1991, which specifically addressed the
question of MRLs from dithiocarbamates. The tolerance was set at 2
ppm for 'tobacco', however, with the following comment: 'The factor
usually applied for the conversion of the weight of fresh tobacco into
the weight of dried tobacco is equal to 5'. De facto, this translates into
a 10 ppm MRL for dithiocarbamates on dry tobacco. The Ordinance
states further that 'the verification of compliance for products based
on tobacco takes place at the moment of putting them on their way of
distribution for consumption'.
Consequently, in the Decrees of 4 July 1992, of 9 August 1995, of 12
August 1995, of 2 April 1996, of 13 January 1997 and of 27 January
1997, two MRLs are stated for most of the pesticides, one for 'green
tobacco' or 'fresh tobacco' and another for 'dried commercial tobacco'.
With one exception (lambda-cyhalothrin), the two numbers always
differ by a factor of 5. Notably, the Decree of 12 August 1995, set the
tolerance for methoprene at 15 ppm, applicable to 'dried commercial
tobacco'.
It is very helpful that the most recent Decree of 22 January 1998
contains an updated complete compilation of all pesticide MRLs on
vegetable crops, including tobacco (Annex 2).
In France, no official legal document such as an Ordinance exists for
the regulation of pesticide residues on tobacco (products). However,
in 1994 a cooperative process was initiated involving the competent
French authorities, the manufacturers of agrochemicals, the growers'
associations and SEITA's Institut du Tabac in Bergerac with the intent
to examine all aspects of

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pesticide use in tobacco production. This is being done with special
attention to the principles of GAP and with the objective, inter alia, to
agree on responsible and practical MRLs for the crop. The first two
pesticides for which such MRLs were set are lambda-cyhalothrin (2
ppm on green leaf and 5 ppm on dried tobacco) and pirimicarb (1 ppm
on green leaf and 2 ppm on tobacco). The process continues and new
MRLs for tobacco are expected to appear.
In Hungary, detailed information on the availability, application and
safe use of pesticides (and crop yield enhancing substances) is
published every year by the Ministry of Agriculture in form of a two-
volume set of books. The 1997 edition (Hungarian Ministry, 1997)
includes MRLs for 27 pesticides on green tobacco leaf and for one
pesticide on dried tobacco (phosphine; 0.1 ppm). For three pesticides
(mancozeb, permethrin and zineb), two different MRLs apply
depending on whether they are used in combination with other
pesticide or not. MRLs for the various dithiocarbamate fungicides
may be as high as, but also lower than, 25 ppm. The MRL for maleic
hydrazide is 20ppm.
The Russian Federation has replaced several older listings of MRLs of
pesticides in foods products, water and soil by a new comprehensive
list of maximum permitted pesticide concentrations in 'environmental
objects' (Russian Federation, 1996). Tobacco is included in the list as
a crop with MRLs stated for 26 individual compounds. It is worth
noting that zineb is considered as the only dithiocarbamate (with an
MRL of 1.0 ppm), together with its degradation product, ethylene
thiourea (with an MRL of 0.02 ppm). In the case of maleic hydrazide,
the MRL is 8.0 ppm a level which is lower than in other countries.
At present, the regulatory status of pesticide residues on tobacco
(products) is rather difficult to assess in the various East European
countries, including those of the former Soviet Union. There is a
considerable degree of uncertainty regarding what regulations do in
fact exist, are retained or are under development. Interestingly, a
number of East European countries seem to be moving in the direction
of greater consistency with existing regulations in West European
countries as well as with European Union regulations for pesticide use
and control.
In the United States, effective 1 July, 1989, imported flue-cured and
burley tobaccos have to comply with MRLs set for 15 pesticides or
combinations of pesticides (USDA, 1989). All pesticides covered in
this regulation are not approved for use on tobacco in the United
States. With two exceptions (ethylene dibromide and formothion), all
of them are chlorine-containing compounds. Several of these
pesticides are also found in the German list of permitted MRLs.
There is considerable similarity between the USA regulation and the
pesticides and their MRLs dealt with in a Standard (Gulf Cooperation
Council, 1995) prepared by the Kingdom of Saudi Arabia for the
member states of the Gulf Cooperation Council. All 14 compounds in
this standard are also found on the US list of pesticides. However,
several of the MRLs are not the same some being higher and some
lower. The MRLs apply to tobacco in the finished product.
Only six agrochemicals have listed MRLs specific for 'tobacco' in the
Food Act 1983 and Food Regulations 1985 (including amendments up
to 1995) of Malaysia (Malaysian Food Act, 1983). Obviously, all of
them are very important for tobacco cultivation and storage:
dithiocarbamates, 25 ppm (expressed as CS2), metalaxyl, 10 ppm,
methoprene, 15 ppm, metolachlor, 0.5 ppm, oxycarboxin, 0.5 ppm,
and permethrin, 10 ppm.
A brief review of the existing regulations for pesticide residues on
tobacco (products) as presented above leaves the impression of a
considerable degree of diversity, regarding both the point of
enforcement and the absolute values of stated maximum residue levels
for a given pesticide.
In certain regulations, MRLs are stated for 'tobacco' without
additional specifications or explanations a situation which generally
leaves the interested reader guessing. On the other hand, the meaning
is obvious in cases where MRLs apply to 'green tobacco' or 'fresh
tobacco' or to 'dried (commercial) tobacco' or 'dried leaf'. The
significance of the term 'finished product' is equally distinct. Some
countries are clear and consistent regarding the point of enforcement,
be it on fresh or cured tobacco leaf (France, United States) or on the
finished product (Germany, Gulf countries). On the other side, Italy
has published MRLs for green tobacco, for fresh tobacco, for dried
commercial tobacco, for finished product and many simply for
'tobacco'.
There is an ongoing discussion regarding the preferred point of
enforcement. It looks like there are good arguments for both
fundamental positions. Setting and enforcing MRLs on the
commodity, tobacco, allows the control of pesticide residues (and the
identification of unacceptable material) close to the origin of the crop
and the source of elevated residue levels. If regulations were strictly
enforced, accountability would be an important factor and the
educational effect on the less responsible could be remarkable. On the
other side, control of marketed product is mandatory as a result of

 Page 260
manufacturers' responsibility for their products and for consumer
protection. In addition, examination of finished product would
automatically reflect the changes (in practice, the decreases) in
pesticide residues which may result from various processing steps and
treatments of the tobacco during manufacturing.
Comparing the absolute values of MRLs of certain pesticides for
different countries reveals considerable heterogeneity. This is
obviously due to a number of reasons. In some cases, MRLs have
been set some time ago and not been re-evaluated since. For others,
factors of agricultural or methodological practicability have been
important. Many of the more recent, extremely low MRLs are the
result of attempts to arrive at the lowest imaginable number without
too much regard for scientific reasoning or agricultural practicability.
For pesticides of importance to the tobacco grower, such situations
must and usually can be addressed using a responsible and
appropriate approach. It looks as if France is about to set a good fresh
example for dealing with the problem.
Analytical Methods for the Determination of Pesticide Residues
A review of the scientific literature reveals that there seems to be no
lack of published methods for the determination of pesticide residues
in foods and consumer products. If, however, stringent criteria of
analytical performance and practical applicability are used, relatively
few of the published methods turn out to be suitable. It is only a recent
requirement for the registration of a pesticide (formulation) in
advanced countries that the applicant (generally the manufacturer)
also provides practicable and reliable methods for the determination
of residues on and in the crops for which the pesticide is approved.
Compilations of methods for the analysis of pesticide residues are
available from several sources. The FDA Pesticide Analytical Manual,
for instance, is divided into a volume with multi-residue methods
(such as screens for organochlorine or organophosphorous pesticides)
and a second volume devoted to methods for single pesticides (US
FDA, 1985). Another source is the manual of the Association of
Official Analytical Chemists, which contains a section dealing with
pesticide concentrations in formulated products and another section
with methods for pesticide residues in foods (Helrich, 1990). The
Deutsche Forschungsgemeinschaft (1991) has released a three-volume
loose-leaf collection of methods dedicated to the determination of
residues from more than 130 active ingredients. This manual is
available in an English language version.
However, only a few of the methods described are immediately
applicable to tobacco. In many cases, however, these manuals offer
good starting points for further methods development. To meet the
urgent requirement for analytical methods which are specific for
tobacco and tobacco products, CORESTA, in one of its working
groups, has developed or examined and improved the analytical
methodology for some of the most important agrochemicals used in
tobacco cultivation. The following six CORESTA recommended
methods (CRMs) are currently available:
Revised method for the determination of dithiocarbamates in tobacco
(CRM # 1ter).
Determinations of organochlorine pesticide residues on tobacco
(CRM # 2).
Determination of maleic hydrazide residues in tobacco (CRM # 4).
Determination of residues of the suckercide flumetralin (Prime Plus,
CGA-41065) on tobacco (CRM # 30).
Determination of residues of the suckercide pendimethalin (Accotab,
Stomp) on tobacco (CRM # 31).
Determination of residues of the suckercide Off-Shoot-T (n-alkanol
mixture) on tobacco (CRM # 32).
Three of these methods have been adopted by ISO as international
standards. The work of CORESTA in this field continues with
considerable urgency.
Effects of Changes in the Regulatory Environment
Tobacco leaf, which is sold on quality and weight, demands high
standards of pest and disease control in its production. This, in the
past, inevitably led to 'insurance' or preventative spraying, where
pesticides are readily available, to realize the maximum potential yield
and quality of farmers' crops. With the current emphasis on
environmental considerations, farmers in all agricultural systems are
encouraged to adopt more rational and targeted approaches for
applying pesticides, rather than the preventative and indiscriminate
applications that have been used in the past.
The tightening of legislation, controlling registration and use of
pesticides in some producing countries, the

 Page 261
plethora of different residue limits that are being imposed at different
stages of tobacco product manufacturing and the ever demanding
criteria that manufacturers are proscribing are placing the traditional
tobacco farmer in an increasingly difficult position. The main
consequences of these changes and actions are now becoming
apparent to the farmers and the world of tobacco leaf trading.
The number of pesticides available to farmers is decreasing as many
are being withdrawn from the market for use on tobacco. The
emphasis placed on monitoring the potential use of some of these
withdrawn, and in some cases banned, pesticides through residue
testing is imposing restrictions on farmers far removed from countries
where such legislation is promulgated.
Agrochemical companies are reluctant to register new products for
use on tobacco due to the large costs and diversity of requirements for
registration in different countries for a relatively small global crop.
The tobacco farmer is, therefore, not always able to take advantage of
the new developments in pesticide chemistry. Furthermore, the
additional testing required from agrochemical manufacturers for
registration for use on tobacco, such as pyrolysis and smoking tests,
can further deter new registrations.
The efficacy of some pesticides has been jeopardized (and reduced)
by the absence of alternative pesticides, with different modes of action
to implement effective 'resistance strategies' against insensitive strains
of fungal pathogens and insects. The use of metalaxyl as the sole
weapon in the arsenal against blue mold in the USA led to a serious
problem with the widespread appearance of strains of the fungus that
were insensitive to the only registered fungicide. Emergence and
restricted registration of mancozeb, an alternative protective
fungicide, and the introduction of a new fungicide, dimethomorph,
have alleviated this particular problem for the time being. However, a
future strategy has to be implemented across the international
boundaries of the USA, Mexico and Central America, if the two
fungicides, metalaxyl and dimethomorph, are to remain effective in
controlling blue mold. An alternating regime of fungicides with
different modes of action has been suggested as a strategy for
combating this problem (Main, et al., 1996).
The widespread use of certain groups of insecticides, particularly the
synthetic pyrethroids and the organophosphates, has led to 'resistant'
strains of aphids and whiteflies becoming widespread and limiting the
use of the favored insecticide. If insecticides with alternative modes of
action are not available, then this situation can very quickly arise and
make effective insect control very difficult. The problem can be
exacerbated in areas where other adjacent crops act as host for
tobacco pests. The extensive use of insecticides with the same mode
of action over wide areas for the growing season of two crops, that do
not coincide but overlap and effectively extend the growing season,
can render an insecticide ineffective in a short period of time. For
example, the tobacco budworm, Heliothis virescens (F.), a major pest
of cotton (and tobacco) in the USA, has developed resistance to nearly
every class of insecticides used in cotton (Elzen, et al., 1992). If the
synthetic pyrethroids had been registered for use on tobacco as well as
cotton, this would have increased the selection pressure in this very
serious situation.
This concern about 'resistance' to pesticides prompted GIFAP to create
four Resistance Action Committees (Fungicide, Herbicide, Insecticide
and Rodenticide Action Committees) in order to understand and
formulate strategies to manage resistance. Surveys carried out by
these Committees have helped illustrate how widespread the problem
is, covering many different species of pests, diseases and weeds on all
world crops, to most of the major pesticide groups (Tomlin, 1997). In
many cases, possibly very serious, multiple resistance is arising when
a pest is insensitive to more than one distinct type of pesticide. The
main strategies involve:
employing all available methods for controlling pests and diseases
including cultural methods, resistant varieties, rotations, crop closed
seasons;
insuring that when using a pesticide the correct dose is always applied
and that label recommendations are followed in terms of the number
of applications;
using different insecticides with different modes of action in sequence
or in mixtures.
The regulatory authorities are aware of the potential problems caused
by pesticide resistance, and pesticide labels now carry warnings and
recommendations if resistance occurs. It has been estimated that in the
USA 30% of the requests to the EPA for Section 18 Emergency
Exemptions have been for the purpose of resistance management
because resistant pest populations have rendered the registered
alternatives ineffective (Lewis & Matten, 1996). The ethylene
bisdithiocarbamate (EBDC) fungicides, mancozeb and maneb,
underwent a special review by the EPA because of their long-term
effectiveness in the field and problems with resistance in other
fungicides. The uses for EBDCs were maintained on numerous
commodities

 Page 262
partly because of the benefits of EBDCs in fungicide resistance
management, for example, EBDCs in combination with benomyl for
controlling apple scab. In 1995, some registrations were obtained for use
on tobacco because of the widespread appearance of metalaxyl
insensitive strains of blue mold.
However, at the same time these regulatory changes, and they are not
restricted to tobacco, have also resulted in many advantages for growers,
manufacturers and consumers:
the virtual elimination of many environmentally and toxicologically
inappropriate pesticides;
resurgence in interest in cultural methods for pest and disease control
through a more integrated approach to crop management;
a tobacco industry with a stewardship campaign promoting the safe and
responsible use of pesticides;
a reduction in the use of pesticides on tobacco which, coupled with a
greater awareness of GAP, will minimize the risk of unacceptable levels
of residues.
These developments have been apparent in all crop protection systems,
and the move towards other forms of control such as cultural practices,
biological control and spraying based on either disease forecasts or
economic thresholds of pest and disease incidence is well developed in
other crops, most notably cotton. This multi-faceted approach to pest and
disease control, commonly used in its simplest forms before the
dependency in some crop systems on pesticides, has become known as
Integrated Pest Management (IPM) and is defined in the FAO (1985)
Code of Conduct on the use of pesticides as:
'Integrated Pest Management means a pest management system that, in the
context of the associated environment and the population dynamics of the
 pest species, utilizes all suitable techniques and methods in as compatible
manner as possible and maintains the pest populations at levels below those
causing economically unacceptable damage or loss.'
This concept has been encouraged by the major pesticide manufacturers
and forms a central philosophy in today's crop protection industry. The
three main elements of IPM are:
(1) identifying the problem such as the pest, disease and weed species;
(2) determining what level of infestation the crop can stand before yield
and quality are compromised while taking into account the cost of the
control measure and the economic threshold;
(3) applying the appropriate means of control which may be a pesticide,
biological control or some form of cultural treatment (see Table 7.4 for
examples).
The most crucial aspect is the training of farmers to monitor their crops
for both injurious and beneficial organisms (scouting) and to understand
the dynamics of the pest or disease build-up. With tobacco, when leaf
Table 7.4 Fundamentals of integrated pest management.
Observations Prevention Intervention
Identification of pest, disease Use of resistant varieties.Hand removal of causal
and weeds. agent if appropriate.
Identification of beneficial Production of healthy The safe use of pesticides
insects/organisms. seedlings for transplants. based on economic
thresholds or accurate
forecasting of epidemics.
Determination of economic Destruction of seedbeds Adherence to pesticide
threshold levels (the level of after transplanting. label recommendations.
infestation below which it is not
necessary to apply a pesticide).
Forecasting infestations and Crop rotations and strict Effective 'resistance
epidemics. 'closed seasons' when management strategies'.
tobacco plants are not in
seedbeds or the field.
Monitoring by scouting fields or Good crop husbandry, Use of biological control
in trapping. e.g. good weed control agents where appropriate.
and removal of debris
after harvesting.

 Page 263
quality is of paramount importance and small blemishes can
significantly reduce the quality and price of the crop, the economic
thresholds in some instances might be low, but IPM is still a relevant
concept and forms a major impetus towards the more rational use of
pesticides.
In tobacco leaf production, formal IPM strategies are actively under
development in a number of countries, for example in the USA, India
and Brazil, but the role of various cultural practices for controlling
pests and diseases in tobacco leaf production is widespread and has
been practised by small-scale farmers for many years. Two examples
of techniques that have been and are contributing to an integrated
approach of pest control and embrace the old and the new are
conventional plant breeding and computer simulation.
The breeding of disease-resistant varieties has been a major objective
of most tobacco breeding programs and has been successful in the US
with many diseases such as Granville wilt, black shank, root knot
nematode (Meliodogyne incognito races 1 and 3), black root rot,
wildfire and a range of virus diseases. In Zimbabwe, varieties with
tolerance to Meloidogyne javanica, white mold, alternaria and angular
leaf spot have been released from Kutsaga Tobacco Research Station.
More recently, a variety that shows resistance to bud worm has been
released in the USA.
The advent of computer simulations of disease epidemics and the
relatively easy access to up to date information on the World Wide
Web through the Internet has provided a new dimension for an IPM
strategy to control blue mold of tobacco in the USA and Central
America in real time (see Chapter 6A). The value of timing a spray
schedule, and thereby potentially reducing the number of spray
applications, based on such information is a major advance for
controlling particular diseases.
Summary
For all agricultural commodities, pesticide residues are an issue of
concern to all participants in the chain from the farmer who grows
crops and wants to achieve maximum returns, to the buyers and
manufacturers who want to insure that their goods and products are
acceptable and can be traded internationally as well as to the
consumer who expects every assurance regarding product safety. The
international diversity and increasing stringency of regulations
controlling the registration and use of pesticides, which have resulted
in an ever decreasing list of products registered for use on tobacco,
have led to a confused situation. This coupled with the absence of any
worldwide accepted standard on pesticide residues has made the
international trading of tobacco leaf and products a complicated and
ever changing business. The outlook, however, does not have to be so
pessimistic. With the ingenuity and dedication of the traditional
tobacco farmer in partnership with the crop protection industry and
support from the tobacco industry, the future could be based on
responsible use of pesticides and stewardship of the environment.
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Elzen, G.W., Leonard, B.R., Graves, J.B., et al. (1992) Resistance to
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 Page 264
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und sonstigen Mitteln in oder auk Lebensmitteln und
Tabakerzeugnissen, of I September 1994; as last amended on 26
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Chapter 8
Leaf Chemistry
8A
Basic Chemical Constituents of Tobacco Leaf and Differences Among
Tobacco Types
J.C. Leffingwell
Leffingwell and Associates
Canton, Georgia, USA
Introduction
In 1960, a little over 200 chemical constituents had been identified in
tobacco leaf of all types and less than 450 had been reported in smoke.
Today, approximately 3000 have been identified and characterized in
tobacco leaf and some 4000 in smoke. Estimates are that the total
number of chemical constituents in leaf exceeds 4000 and there are
over 6000 in tobacco smoke. It is not the purpose of this section to
comprehensively review all of the known constituents, but rather to
provide an insight into the known composition and chemistry of
tobacco types that impact tobacco quality and differentiate tobacco
types. Emphasis will be placed on the major tobaccos utilized
commercially: Virginia (fluecured), air-cured (burley and cigar) and
Oriental.
The physical and chemical properties of leaf tobacco are influenced
by genetics, agricultural practices, soil type and nutrients, weather
conditions, plant disease, stalk position, harvesting and curing
procedures. A change in any of these factors can markedly alter the
chemical composition of leaf and thus affect smoking quality (Tso,
1972; Tso, 1990).
It is now generally accepted that the metabolic carbon-nitrogen
balance in living plants is due to continuing transformations based on
the Krebs tricarboxylic acid cycle. In the Krebs cycle, carbon dioxide
from air is assimilated through photosynthesis in the tobacco leaf
while inorganic nitrogen (nitrate and/or ammonia) is assimilated
through the roots from the soil. Soil nitrate is converted to ammonia
which is utilized in the Krebs cycle to form amino acids which serve
as a nitrogen pool for the formation and transformation of a multitude
of nitrogenous chemicals important in the development of aroma and
flavor quality. Dawson (1952) has suggested a concept based on the
Krebs cycle to account for inherited and culturally induced variations
in gross tobacco composition. Using this concept, he rationally
suggested that for tobaccos where the nitrogen supply is abundant,
such as in cigar and burley tobacco production, there should be an
abundant formation of protein, amino acids and nicotine. For Oriental
tobacco, where growth is maintained with limited supplies of nitrogen
nutrients and water, there is an accumulation of acetate in the Krebs
cycle resulting in the biosynthesis of terpenoids via mevalonic acid as
well as a higher production of carbohydrates, 'aromatic' acids and
resins at the expense of nitrogen constituents. Fluecured tobacco is
intermediate in that the phytochemistry during the plant's life cycle is
balanced by a moderate supply of nitrogen which is depleted as the
plant reaches maturity.
Examination of representative analyses of the major cigarette tobacco
types as presented by Harlan and Moseley (1955) (Table 8.1) provides
an overview of the major differences in aged fluecured, burley,
Maryland and Oriental tobaccos.
Although average reducing sugar content in fluecured and Oriental
cigarette tobaccos today rarely runs as high as reported in Table 8.1,
the basic analytical trends still remain valid. Thus, in air-cured burley,
Maryland and cigar tobaccos the carbohydrates have been virtually
depleted via metabolism of the living cells, whereas the protein and a-
amino nitrogen are obviously higher than in fluecured or Oriental
tobaccos. Conversely, the fluecured and Oriental

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Table 8.1 Composition of cigarette tobaccos: representative analyses of cigarette
tobaccos (leaf web after aging, moisture-free basis).
Component
(%)1 Flue-cured, Burley, type Maryland, type Oriental2
type 13 31 32
Total volatile bases as 0.282 0.621 0.366 0.289
ammonia
Nicotine 1.93 2.91 1.27 1.05
Ammonia 0.019 0.159 0.130 0.105
Glutamine as ammonia 0.033 0.035 0.041 0.020
Asparagine as ammonia 0.025 0.111 0.016 0.058
a-Amino nitrogen as 0.065 0.203 0.075 0.118
ammonia
Protein nitrogen as 0.91 1.77 1.61 1.19
ammonia
Nitrate nitrogen as NO3 trace 1.70 0.087 trace
Total nitrogen as ammonia 1.97 3.96 2.80 2.65
pH 5.45 5.80 6.60 4.90
Total volatile acids as 0.153 0.103 0.090 0.194
acetic acid
Formic acid 0.059 0.027 0.022 0.079
Malic acid 2.83 6.75 2.43 3.87
Citric acid 0.78 8.22 2.98 1.03
Oxalic acid 0.81 3.04 2.79 3.16
Volatile oils 0.148 0.141 0.140 0.248
Alcohol-soluble resins 9.08 9.27 8.94 11.28
Reducing sugars as 22.09 0.21 0.21 12.39
dextrose
Pectin as calcium pectate 6.91 9.91 12.41 6.77
Crude fiber 7.88 9.29 21.79 6.63
Ash 10.81 24.53 21.98 14.78
Calcium as CaO 2.22 8.01 4.79 4.22
Potassium as K2O 2.47 5.22 4.40 2.33
Magnesium as MgO 0.36 1.29 1.03 0.69
Chlorine as Cl 0.84 0.71 0.26 0.69
Phosphorus as P2O5 0.51 0.57 0.53 0.47
Sulfur as SO4 1.23 1.98 3.34 1.40
Alkalinity of water-soluble 15.9 36.2 36.9 22.5
ash3
1 In % except for pH and alkalinity.
2 Blend of Macedonia, Smyrna, and Samsun types.
3 Milliliters of 1 N acid per 100 g tobacco.
Source: Harlan and Moseley (1955).

tobaccos possess significant amounts of reducing sugars (which are

virtually absent in the air-cured tobaccos) and lesser amounts of
protein and a-amino nitrogen. Substantial changes in the chemical
composition of tobacco leaf occur following harvest and during
subsequent processes.
Carbohydrates: Starch, Sugars, Sugar Esters, Cellulose, and Pectin
a
Starch and Sugars
In flue-cured tobacco, respiration in the primed leaf is arrested by the
controlled desiccative dehydration during flue-curing which causes
enzyme inactivation. Nevertheless, considerable change has occurred
(Table 8.2) in the leaf (during the period after priming and early stages
of curing) as starch loss occurs via enzymatic hydrolysis with a
concomitant increase in reducing sugar (Bacon, et al., 1952). This is
illustrated for the starch depletion during cure of Virginia tobacco
(Fig. 8.1) and the concurrent generation of reducing sugars (Long &
Weybrew, 1981) (Fig. 8.2).
In burley tobacco the starch accumulation during growth is only about
25% the amount in Virginia tobacco (Weybrew & Hamann, 1977) and
this is nearly depleted completely during the catabolic respiration of
the plant while air-curing leaving negligible sucrose, and reducing
sugars in the cured leaf.

 Page 267
Table 8.2 Changes in composition of Virginia
tobacco during the flue-curing process (% of dry
weight).
Constituents Green Yellowed Cured
Starch 29.30 12.40 5.52
Free reducing sugars 6.68 15.92 16.47
Levulose 2.87 7.06 7.06
Sucrose 1.73 5.22 7.30
Crude fiber 7.28 7.16 7.24
Total nitrogen 1.08 1.04 1.05
Protein nitrogen 0.65 0.56 0.51
Nicotine 1.10 1.02 0.97
Ash 9.23 9.24 9.25
Calcium 1.37 1.37 1.37
Oxalic acid 0.96 0.92 0.85
Citric acid 0.40 0.37 0.38
Malic acid 8.62 9.85 8.73
Resins 7.05 6.53 6.61
Pectinic acid 10.99 10.22 8.48
pH of tobacco 5.55 5.64 5.55
Source: Bacon, et al. (1952).
 Fig. 8.1
Change in lamina starch during flue-curing of
Virginia tobacco (adapted from Long &
Weybrew, 1981).
Starches are generally polymers of two polysaccharides: amylose and
amylopectin. Corn starch has approximately 27% amylose and 73%
amylopectin, whereas tobacco has been found to have approximately
23% amylose and 77% amylopectin (Johnstone & Plimmer, 1959).
The amylose portion is estimated to have a chain length of 40 to 47
anhydro-glucose units, while the amylopectin has about 26 glucose
units.

Fig. 8.2
Change in lamina reducing sugars during
flue-curing of Virginia tobacco (adapted
from Long & Weybrew, 1981).
b
Sugar Esters
The first report of sugar esters in Oriental tobacco came in 1970 with
the isolation, structure elucidation and synthesis of 6-0-acetyl-2,3,4-
tri-0-[(+)-3-methylvaleryl]-beta-D-glucopyranose (a glucose tetraester)
by Schumacher (1970). This and the more predominant sucrose
tetraesters (STE) of lower carboxylic acids (Fig. 8.3) are now
considered to be some of the most important aroma precursors
responsible for Oriental flavor (Leffingwell & Leffingwell, 1988).
In 1981, Severson, et. al., found that the cuticular waxes of a tobacco
budworm resistant tobacco contained a series of STE which are the
probable precursors of 6-0-acetyltriacylglucopyranosides (glucose
tetraesters (GTE)) isolated from Oriental tobacco (Severson, et al.,
1981; Severson, et al., 1985a; Severson, et al., 1985b).
Einolf and Chan have quantified the accumulation of STE for Coker
319 (Virginia), Kentucky 14 (burley) and Smyrna (Oriental) tobaccos
utilizing flue-curing, air-curing and sun-curing respectively to mimic
the field-curing practices for each tobacco type (Einolf & Chan,
1984). The graphic representation (Leffingwell

 Page 268

Fig. 8.3
Sugar esters present
in Oriental tobacco.
& Leffingwell, 1988) (Fig. 8.4) shows that accumulation of STE for
Oriental and burley is greater than for Virginia tobacco, with the
Oriental and burley maxima occurring at full maturity.
It has been demonstrated that the tetraesters of Oriental STE tended to
have higher amounts of the
 Fig. 8.4
Sucrose tetraesters in tobacco types (adapted from Einolf
& Chan, 1984).
more aromatic C5 and C6 carboxylic acids (Severson, et al., 1985b).
From a smoke aroma and flavor standpoint, it is interesting that GTE
and STE readily release free carboxylic acids on thermolysis while
totally esterified sucrose and glucose esters (such as glucose
pentaisovalerate and the sucrose octaesters) do not easily release their
acid moieties on thermolysis.
c
Glucosides
The suggestion that nonvolatile aroma precursors of tobacco existed
in the form of glucosides was made by the observation that materials
such as 3-hydroxydamascone and 3-keto-ionol were released from
tobacco by hydrolysis (Green, et al., 1980). Very little information on
the hydrolyric release of aroma compounds from fruits and other plant
materials by either the acid or enzymatic hydrolysis of glycosidically
bound terpenoid and norterpenoid materials was known prior to the
mid-1970s. Today, it is clear that many plants, including tobacco,
which have sugars (mono-and disaccharides) generate nonvolatile
materials with an inter-sugar link to aroma materials arising from the
oxidative and metabolic transformation of carotenoid pigments
(Williams, 1993). Many tobacco identical aroma compounds have
now been found in fruits, wines and other products, primarily bound
as glucosides. Heckman, et al. (1981) demonstrated that a series of
tobacco aglucone isolates are significantly increased by enzymatic
hydrolysis of glucosides from Virginia tobacco (Table 8.3).
The commercial enzymatic hydrolysis of tobacco scrap and dust has
been used to generate tobacco aroma materials from the glucosidically
bound precursors in tobacco by one of the major flavor companies.
The use of glucosides for flavoring tobacco has been patented
(Anderson, et al., 1976), and commercial use of the glucoside of ethyl
vanillin (Herron, 1987) (an artificial flavor about three times more
powerful than vanillin) for improving sidestream aroma was
employed at one time in the US in cigarettes.
d
Cellulose
It has been estimated (Green, 1977) that cigarette blends contain
approximately 10% of cellulose and hemicellulose, which corresponds
closely to the values obtained for a series of flue-cured varieties
(Collins & Legg, 1977). However, it must be noted that the tobacco
stalk, stem and mid-rib portion of the leaf are much higher in cellulose
than the lamina portion of the leaf.

 Page 269
Table 8.3 Aglucone increase from enzymatic hydrolysis of glucosides in Virginia
tobacco (mg/100 g).
Aglucone Virginia Enzyme- Percent
control treated increase
3-Methylbutanol 0 104 N/A
Benzyl alcohol 1815 3167 74
2-Phenylethanol 887 1744 97
2-Methoxy-4-vinylphenol 498 792 59
4-Vinylphenol 1023 1255 23
3-Hydroxyldamascone 2469 2697 9
4-Vinylsyringol 1615 2850 77
3-Keto-a-ionol 7257 10993 52
4-(4-Hydroxy-2,6,6-trimethyl-1-cyclohexen-1- 1783 1805 1
yl)-3-butyn-2-ol
3-Hydroxy-5,6-epoxy-b-ionol 248 698 181
4-(3-Hydroxybutenylidene)-3,5,5- 426 1146 169
trimethylcyclohex-2-en-1-one
3-(2-Hydroxyethyl)-phenol 155 1207 679
6-Hydroxy-3-keto-a-ionone 67 109 63
Vomifoliol 416 701 69
Source: Heckman, et al. (1981).

Cellulose is a polysaccharide of the general formula (C6H10O5)n which

usually consists mainly of glucose units. Acid hydrolysis of cellulose
fractions from Japanese Bright (Virginia) tobacco leaf yielded only
glucose from one fraction and mainly galactose from another, while the
stem cellulose yielded mainly arabinose and glucose (Johnstone &
Plimmer, 1959).
Normally, high cellulose content in a tobacco blend is a negative to
smoking quality in that it tends to impart a sharp stinging harshness and
a 'burnt paper' odor to the smoke. The cellulose content of stems plays
an important part in the manufacture of reconstituted tobacco because
the fiber provides structural strength. The degree of polymerization
(DP) of cellulose from leaf lamina varies considerably from that of mid-
rib and stems (DP of 11001650 for lamina versus 16001800 for leaf
mid-ribs). These values are considerably lower than that found for wood
fiber (DP of about 3000). In addition, the DP for Virginia tobacco tends
to be lower than for air-cured tobaccos (Stedman, 1968). Pyrolysis
studies on the generation of formic acid from tobaccos also suggest that
Virginia cellulose may have a shorter mean chain length than burley
(Fenner, 1988).
As tobacco stems play an important role in the modern economics of
tobacco manufacture, being employed as both cut rolled expanded stems
and as a cellulose source in reconstituted tobacco, it is noted that
Virginia or flue-cured stems are preferred to air-cured stems for their
smoking quality. This may be due to the difference in cellulose
polymerization, as well as other materials present (such as higher nitrate
(and combustibility)) in air-cured stems.
e
Hemicellulose
Hemicelluloses are a group of polysaccharides found in the cell walls of
all plants in association with lignin. Whether lignin and hemicellulose
are bonded or the hemicellulose is mechanically entrapped by lignin is
unclear (Anon, 1989). The hemicellulose of Japanese Bright (Virginia)
tobacco leaf was separated into two fractions which on hydrolysis gave
mainly arabinose, galactose and glucose, along with lesser amounts of
xylose (Johnstone & Plimmer, 1959). Hemicellulose is subject to
extreme variations as its composition can change with growth and
maturation and is influenced by cultural and environmental factors.
f
Pectin
The tobacco pectins are the 'glue' that holds tobacco leaf together. Pectin
comprises 6 to 12% of tobacco weight and as such contributes
importantly both to the structural stability of leaf and to pyrolysis
products that contribute to the smoke chemistry.
The pectin molecule is basically a polymeric carbohydrate comprised of
1,4-linked a-D-galactouronic acids with various degrees of methylation
of the carboxylic acid groups. Dispersed within the polygalacturonic
acid backbone are neutral sugar moieties such as galactose, rhamnose
and arabinose.
Bokelman and coworkers (Sun, et al., 1987) found that the main pectic
polysaccharide fraction from tobacco has a backbone of 1,4-linked a-D-

 Page 270
galactopranosyluronic acid interspersed with 2-linked L-
rhamnopyranosyl residues. Approximately 22% of the galacturonic
acid was methylated. The main pectic chain was determined to be
branched at carbon-4 of rhamnose with the neutral side chains
containing terminal and 4-linked b-D-galactopyranosyl and a-L-
arabinofuranosyl residues.
Lauterbach and Moldoveanu (1991) determined the degree of both
methylation and amidation by pyrolysis gas chromatography-mass
spectrometry (GC-MS) for the soluble pectic fractions of commercial
grades of cured Virginia, burley and Oriental tobaccos with the
following results:

Tobacco Methoxylation Amidation

(%) (%)
Virginia 20 24
Burley 4 34
Oriental 12 16

It has long been known that the pectin fractions of tobacco are
modified chemically by curing procedures (especially air-curing and
cigar fermentation where ammonia can interact with the pectic
substances present).
From a commercial aspect, pectins play an important role in the
manufacture of reconstituted tobacco. One of the two types of major
commercial reconstituted tobacco processes involves the treatment of
a tobacco slurry with diammonium phosphate to solubilize the
tobacco pectin. When the tobacco slurry is then cast onto a hot metal
belt, a portion of the ammonia is volatilized, 'setting' the tobacco
pectin to 'glue' the reconstituted sheet together. Because the ammonia
generated plays an important role in natural tobacco flavor formation,
this reconstituted process not only provides a means of extending
tobacco utilization, it also provides a superior reconstituted tobacco
from a smoking quality standpoint by increasing the flavorful
pyrazines. The role of ammonia in tobacco will be discussed below.
Carbohydrates and Smoke Chemistry
Carbohydrates in the forms previously described can account for 40 to
50% of tobacco weight and contribute significantly to smoking
quality. As mentioned, the sugar esters are a major contributor to the
Oriental tobacco flavor, in that they thermally release the potent lower
fatty acids characteristic of Turkish smoke. In addition, it has been
noted that of the 147 pyrolysis products reported from cellulose,
starch, mono-and di-saccharides and pectic substances that 121 have
been found in tobacco smoke (Heckman, et al., 1981).
Sensory Role of Carbohydrates, Ammonia and Nicotine PH of
Tobacco Smoke
Pyrolysis of carbohydrates in tobacco smoke has long been known
empirically to change the sensory impression of certain types of
tobaccos. Even before cigarettes became popular, sugars were added
to air-cured tobaccos used for pipe smoking to mitigate the harshness
of nicotine in the smoke. And, today, as 50 years ago, tobaccos are
purchased and blended based on certain chemical criteria (sugar and
nicotine content) as well as the more subjective criteria of appearance
and smoke flavor. In addition, it is a common practice to add sugars
(usually invert sugar (Glucose + fructose) or partially inverted sugar
which still contains some sucrose) in American blend cigarettes.
Sugars are known to 'balance' the smoke flavor, primarily by
modifying the sensory impact of nicotine and other tobacco alkaloids
(Leffingwell, 1976).
In the section on nitrogenous constituents of tobacco, the interaction
of amino acids and ammonia in the development of important
compounds in both Virginia and air-cured tobaccos (including amino-
sugar compounds) will be discussed.
However, at this point a brief discussion on the effect of the major
acid forming constituents (carbohydrates) of tobacco smoke and the
interaction with the major basic substances of tobacco and smoke
(nicotine and ammonia) is in order.
Fenner (1988) showed the striking differences in the generation of
both formic acid (Fig. 8.5) and ammonia (Fig. 8.6) from Virginia and
burley tobaccos on pyrolysis over the temperature range encountered
in a cigarette.
In Fig. 8.5, we see the generation of formic acid in Virginia tobacco to
be far greater than for burley with the large transition at 190°C being
due to simple sugars, while the transition at 250°C is due to pectin
plus hemicellulose and the transition at 310°C. is due to cellulose.
Conversely, Fig. 8.6 demonstrates that burley tobacco generates
significantly more ammonia than Virginia. The ammonia transition at
190°C. may simplistically be considered as a labile or exchangeable

 Page 271

Fig. 8.5
Thermal generation of formic acid (adapted from
Fenner, 1988).

Fig. 8.6
Thermal generation of ammonia (adapted from
Fenner, 1988).
ammonia source (e.g. ammonium salts), while the 350°C. transition is
amine derived and the 550°C transition is amide-derived ammonia.
Acid forming constituents (carbohydrates) and basic constituents
(ammonia) can have a significant effect on smoke pH. Figure 8.7
shows the cumulative effect of successive puffs on the pH of water
through which smoke has been passed. In a study of over 150 brands
of cigarettes, Elson, et al. (1972), concluded that brands with low
sugar content generated more alkaline smoke than those with high
sugar content (which become progressively more acidic).
As nicotine is a major volatile base present in

Fig. 8.7
Cigarette smoke pH (cumulative puffs) (adapted from
Elson, et al., 1972).
cigarette smoke, the pH of smoke plays an exceedingly important role
on the sensory impression imparted.
The higher ammonia level of air-cured tobacco containing low or
negligible sugar content would presumably have two major effects. It
has an effect on the ratio of nicotine/nicotine salts delivered on
smoking relative to high sugar and/or more acidic tobacco types
wherein the nicotine and other alkaloids are complexed (more) as salts
in the smoke.

Since the pH of smoke in air-cured tobacco is considerably more

alkaline than flue-cured or Oriental, the ratio of nicotine base to
nicotine salts increases. This causes the sensory and physiological
perception of increased nicotine strength (and harshness). It should
also be noted that the pH of air-cured tobacco smoke increases with
succeeding puffs from such cigarettes. Accordingly, the increased
alkalinity of straight air-cured cigarettes renders them virtually
unacceptable to nearly all smokers as the high smoke pH imparts an
alkaloid harshness with a flavor distortion which can be unpleasant.
Conversely, many American blend cigarette smokers find the 'acidic'
smoke of straight Virginia cigarettes to be

 Page 272
unbalanced. 'Nicotine alone does not determine smoking flavor'
(Leffingwell, 1976).
Nitrogenous Constituents
The data in Table 8.4 for air-cured cigar tobacco show the gross
chemical changes in nitrogen compounds during air-curing
(Frankenburg, 1946; Frankenburg, 1950).
Table 8.4 Changes of nitrogenous compounds in air-
cured cigar tobacco (percent of harvested dry
weight).
Primed leaf Stalk cured
Type of Before After Before After
nitrogen curing curing curing curing
Total 5.61 5.34 4.70 3.80
Protein 3.69 1.65 3.80 1.85
(insoluble)
Soluble 1.92 3.69 0.90 1.95
Amino 0.23 0.80 0.15 0.15
Ammonia plus 0.15 1.07 0.05 0.80
amide
Alkaloid 0.35 0.32 0.40 0.40
Nitrate 0.63 0.77 0.20 0.25
Remainder 0.56 0.73 0.10 0.35
Source: Frankenburg (1946).

In general, there occurs some loss in total nitrogen, a reduction in

protein via hydrolysis to amino acids, and formation of amino acid
amides (as reflected by an increase in soluble nitrogen). Changes
occurring in burley tobacco curing (Hamilton, 1974; Hamilton &
Lowe, 1978) are similar to those in stalk-cured cigar tobacco.
Flavor quality for both flue-cured and burley tobaccos is dependent on
a variety of complex interacting factors related to genetics,
agricultural practices, soil types, nutrients, weather conditions, plant
disease, stalk position as well as harvesting and curing practices. In
general, elevated levels of proteins and amino acids appear to
contribute negatively to smoke flavor. This is particularly true of
burley tobaccos grown in cool moist climates with insufficient
sunshine which are over-fertilized, harvested and air-cured
prematurely. In such cases the normal catabolic changes during
senescence and curing which result in the enzymatic hydrolysis of
fraction 1 protein to amino acids (which then can undergo further
catabolic changes) are at least partially arrested. Such immature
tobaccos possess a proteinaceous aroma on smoking which is
unpleasant. Even secondary fermentation of such poor tobaccos often
cannot provide sufficient improvement to make them usable. By
comparison to normal burley, in which approximately 50% of total
protein undergoes hydrolysis to amino acids, such poor tobaccos may
exhibit less than 15% protein hydrolysis during air-curing (Heinzer,
1986). Normal air-curing of fully mature burley shows a rapid drop in
protein (Fig. 8.8) (Hamilton & Lowe, 1978; Long & Weybrew, 1981)
with a concomitant initial increase in free amino acids for the first few
days in cure (Fig. 8.9) (Burton, et al., 1983). Then, there is a decrease
and finally a leveling off in total free amino acid concentration. The
decrease in free amino acid content after the fifth day of air-curing is
due to both metabolic deamination and decarboxylation as well as
nitrogen translocation. Even though a slight decrease from a
maximum in total free amino acids occurs during the cure, some
individual amino acids show a net decrease while others have a net
increase (Burton, et al., 1983). It should be noted that the ammonia
level of air-cured tobacco rapidly increases during this period (Fig.
8.10) (Burton, et al., 1983).
The role of ammonia on smoke pH and nicotine perception has been
discussed above, but ammonia also plays an important role in the
formation of aroma and taste compounds.
It is important to remember that, before curing, air-

Fig. 8.8
Protein decreases during burley air-curing
(adapted from Long & Weybrew, 1981).

 Page 273

Fig. 8.9
Total free amino acid changes during burley
air-curing (adapted from Burton, et al., 1983).

Fig. 8.10
Ammonia increases during burley air-curing
(adapted from Burton, et al., 1983).
cured burley or cigar tobaccos may contain 3 to 4% protein nitrogen,
while flue-cured tobacco contains only 15 to 20% of that amount. In
addition, about 50% of the protein in burley may undergo hydrolysis
during air-curing while only about 20% of the protein is hydrolyzed in
curingcured tobaccos. Thus, burley tobacco possesses a relative
abundance of free amino acids compared to flue-cured tobacco.
Representative free amino acid analysis (Table 8.5) of high
Table 8.5 Free amino acids in high quality
smoking grade blends (mg/g).
Amino acid Flue-cured Burley
Aspartic acid 0.13 7.84
Threonine 0.04 0.43
Serine 0.06 0.17
Asparagine 1.12 10.30
Glutamic acid 0.10 1.78
Glutamine 0.82 0.38
Proline 4.11 0.45
Glycine 0.02 0.14
Alanine 0.32 0.35
Valine 0.06 Trace
Isoleucine 0.06
Leucine Trace 0.10
Tyrosine 0.68 0.84
Phenylalanine 0.24 0.50
Lysine 0.03 0.33
Histidine 0.11 0.45
Arginine 0.26
Tryptophan 0.50
Source: Leffingwell (1976).

smoking quality blends of flue-cured and burley tobaccos with similar

nicotine content illustrates this dramatically (Leffingwell, 1976). In
burley, we find the main free amino acids present to be aspartic acid,
asparagine and glutamic acid, while for flue-cured, proline, asparagine
and glutamine are dominant. In total, 43 amino acids have been found
in tobacco leaf.
Cigarette tobaccos are almost never utilized in consumer products
unless they have undergone a period of aging ranging from 12 to 60
months. Of course, inventory control is an important factor requiring
maintenance of adequate stocks to allow periodic blend changes to
occur; but, freshly cured and redried leaf is not used primarily because
the smoke changes dramatically from a raw, somewhat irritating and
disagreeable taste to a smoother, more rounded flavor during the aging
process. Aging is ordinarily carried out at ambient temperatures and
about 12% moisture. In areas with relatively long-term high ambient
temperatures the desired aging effects are produced in shorter periods
than in colder climates (Bates, et al., 1974), and it is common for
companies located in cold climate regions to store and age their
tobaccos in southern locales. The role that nitrogen compounds play
in the improvement of tobacco flavor quality with aging is important
and the reasons for this will become apparent.
The role of certain amino acids in flue-cured tobacco

 Page 274
will be briefly discussed. Protein hydrolysis with formation of free
amino acids occurs also at the yellowing and early stages of drying
during flue-curing, although flue-cured tobacco contains only about
20 to 25% of the total amount of protein/amino acids as burley
tobacco. The major amino acid in flue-cured tobacco, proline, seems
to be an anomaly in that as much as a 25-fold increase has been
observed during the curing schedule for both flue-cured and air-cured
(Maryland) tobaccos (Weybrew, et al., 1966). This appears to be
greater than can be accounted for by proteolysis of fraction 1
protein a hypothesis (Hamilton & Lowe, 1978) being that proline is
partially formed by a metabolic conversion of the pyrrolic portion of
chlorophyll (which is rapidly decreasing during this stage).
In flue-cured tobaccos, proline reacts with the reducing sugar,
glucose, to form l-deoxy-l-(L-prolino)-D-fructose (Noguchi, et al.,
1971), an Amadori compound, which along with other amino-sugars
comprises as much as 1.5 to 2.0% of the dry aged tobacco weight.
This compound is significant because it has been shown to undergo
low temperature degradation primarily to the probable precursor (2,3-
dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP)) of the
important volatile flavorant, maltol, found in Virginia tobacco
(Leffingwell & Leffingwell, 1988). It has been shown that the
smoking properties of DDMP are identical to that of maltol
(Leffingwell, 1976).
Figures 8.11 and 8.12 show the change in two of the major Amadori
compounds in Virginia tobacco grown in Japan over a 4-year aging
period (Noguchi, et al., 1971). The major Amadori compounds
present tend to show a gradual increase peaking at about 2 years and
then a decrease. During the same period, the amino acid content of the
tobaccos shows decreasing values each year. Of course, it is also
known that the flavor of Virginia tobacco improves with aging, it
rarely being used for cigarettes prior to 12 to 14 months aging, and
preferably 18 to 24 months, which interestingly corresponds quite
well to these graphs. Amadori compounds are also known to generate
a number of flavorful pyrazines and pyrroles on pyrolysis.
Amino acids as well as ammonia interactions with sugars or carbonyl
compounds can form a variety of chemical entities. For example, a
number of groups have reported the isolation of fructosazines and
deoxyfructosazines from Virginia tobacco (Shigematsu & Kitami,
1978; Heckman, et al., 1981) and these are known to be formed by the
interaction of ammonia with glucose and fructose under weakly acidic
conditions. On pyrolysis, these polyhydroxypyrazines generate
products such as water, acetic acid, acetol, furans,

Fig. 8.11
Changes in Amadori compounds (prolinofructose)
during aging of flue-cured tobacco (adapted from
Noguchi, et al., 1971).
 Fig. 8.12
Changes in Amadori compounds (alanino-fructose)
during aging of flue-cured tobacco (adapted from
Noguchi, et al., 1971).

 Page 275
pyrroles and a number of simple pyrazines (including the major
tobacco leaf pyrazines, 2,5-and 2,6-dimethylpyrazine,
trimethylpyrazine and tetramethylpyrazine) (Heckman, et al., 1981).
Although pyrazines as a class represent a very small percentage of
tobacco weight, they are known to be very important to the flavor and
aroma of roasted foodstuffs and tobacco.
Bright and co-workers demonstrated that the weight percentages of
five amino acids were significantly reduced by heat treatment, while
the roasting of tobacco caused dramatic increases in pyrazine levels
(especially those high in ammonia or ammonia precursors) under
conditions where percent of nicotine remained constant (Tables 8.6
and 8.7) (Bright, et al., 1975).
Table 8.6 Change in amino acid levels
of tobacco after roasting (complete
cigarette blend).
Amino acid Before (%)After (%)D%
Aspartic acid 1.39 0.82 -41
Proline 0.65 0.31 -52
Lysine 0.32 0.10 -69
Histidine 0.24 0.16 -33
Arginine 0.16 0.06 -62
Source: Bright, et al. (1975).

Table 8.7 Effects of roasting on

dimethylpyrazine concentration.
Concn before Concn after
roasting (ppb) roasting (ppb)
Virginia 0.1 0.3
Turkish 0.1 0.8
Burley 0.1 460
Reconstituted 0.1 200
leaf
Tobacco samples were shredded, without
additives and masted for 4 hours at 120°
±3°C.
Source: Bright, et al. (1975).

Wahlberg, et al. (1977) have examined the chemical changes

occurring during the flue-curing and aging of Virginia tobacco. One of
the observations was that a number of pyrazines, pyrroles and
pyridines increased during the flue-curing and aging process (Enzell,
et al., 1977). As pyrroles, pyridines and pyrazines can arise in
tobaccos by a number of chemical pathways ranging from the heating
or aging of proteins and amino acids
to the interaction of amino acids and/or ammonia with sugars or
carbonyl constituents, suffice it to say, no single pathway is involved
and that this complex transformation is dependent on a variety of
factors and chemical mechanisms (Leffingwell & Leffingwell, 1988).
The generation of certain pyridines (e.g. methyl nicotinate, 3-
cyanopyridine and 3-acetylnicotine) can be derived from degradation
of nicotine.
Tobacco Alkaloids
The major tobacco alkaloids are nicotine, cotinine, nornicotine,
myosmine, nicotyrine, anabasine and anatabine. This subject is
covered in greater detail elsewhere in this book (Chapter 8B) and will
not be discussed here except to note that nicotine can range in
concentration from 0.5 to 8% in the major cultivated tobacco species,
N. tabacum and N. rustica. The interaction of smoke pH and the
sensory perception of nicotine is discussed above.
Plastid Pigments
The major pigments of tobacco are the chlorophylls and carotenoid
pigments, plastid pigments found in a wide variety of plants. The
chlorophyll and carotenoid contents and changes during the latter
stages of growing (and the curing processes) have been studied in
significant detail for both burley and Virginia tobaccos.
a
Chlorophylls
As shown in Fig 8.13 for burley tobacco, the major green pigments
are chlorophyll-A and chlorophyll-B, both of which decrease as the
tobacco reaches maturity, proceeds into senescence and continues to
decrease during barn curing (Burton & Kasperbauer, 1985). Studies
on Virginia tobacco show similar trends as the tobacco reaches
maturity (Court & Hendel, 1984).
Leaf color in the field as well as for cured leaf is considered to be
important in judging the quality of leaf and this has been extensively
studied (Tso, 1972; Tso, 1990).
b
Carotenoid Pigments
The major carotenoid pigments in tobacco are lutein, b-carotene,
neoxanthin and violaxanthin. In addition to being major color
pigments (red-orange to yellow), the

 Page 276

Fig. 8.13
Changes in chlorophyll content of burley
tobacco during air-curing (adapted from
Burton & Kasperbauer, 1985).
carotenoids are the precursors to many of the volatile aroma
components of tobacco.
Figure 8.14 shows the decrease in carotenoids during maturation,
senescence and curing for burley tobacco (Burton & Kasperbauer,
1985). Unlike the chloroplastid pigments, the carotenoids do not
undergo as extensive degradation (metabolism). Lutein and carotene
undergo about 65% degradation in burley.
 Fig. 8.14
Changes in carotenoid pigments of burley tobacco
during air-curing (adapted from Burton &
Kasperbauer, 1985).
More extensive degradation of lutein has been reported for Virginia
tobacco (Court & Hendel, 1984; Court, et al., 1993).
Tobacco Isoprenoids
Wahlberg and Enzell (1987) have comprehensively reviewed the
subject of tobacco isoprenoid constituents, therefore only comments
on some of the more important materials will be included in this
section.
a
Degraded Carotenoids
A large number of carotenoid metabolites are encountered in tobacco.
The degradation process of the carotenoid pigments in tobacco has
been hypothesized to be probably both enzymatic and autooxidative in
nature. As indicated in Fig. 8.15 for carotene, the carotenoid chain
may be cleaved in a number of locations resulting in a variety of C9 to
C13 chemicals, many of which possess important aroma properties.
Table 8.8 (Wilson, et al., 1982; Leffingwell & Leffingwell, 1988)
illustrates the quantitative amounts found for a number of important
nor-carotenoids for various tobacco types.
The degradation of both lutein and b-carotene by both photo-oxidation
and high pressure oxidation (at elevated temperature) has been shown
to generate
 Fig. 8.15
Carotenoid consituents of burley tobacco.


 Page 277
Table 8.8 Major tobacco carotenoid degradation
products.
Compound Virginia Burley Oriental
(ppb) (ppb) (ppb)
112 29 33

Isophorone
47 17 42

4-Ketoisophorone
32 4 43

Safranal
150 26 73

Dihydractinodiolide
139 25 34

Oxoeudalane
47 27 73

Dihydro-
oxoeudalane
+ + +

a-Ionone
+ + +

b-lonone
+ NR NR

3-Keto-a-ionone
+ 4 +
3-Keto-a-ionol
51 31 23

3-
Ketodihydroionone
168 42 39

Damascenone
24 11 +

3-Hydroxy-b-
damascone
1272 248 415
Megastigmatrienones
(5 lsomers)
NR + NR

4-Keto-b-ionone
NR = not reported; + = present, but quantitation
not available.
Table adapted from Wilson, et al. (1982) and
Leffingwell & Leffingwell (1988).

many of the same nor-carotenoid components found in tobacco (Heij,

et al., 1992).
b
Acyclic Isoprenoids and nor-Derivatives
Solanesol (a C45 polyprenol) is a major component of tobaccos,
generally ranging in quantity from 0.4 to 4%. While this polyprenol
was first isolated from tobacco, it is now considered to be a ubiquitous
leaf component in the plant kingdom. The progression of solanesol
change during growth and air curing for middle stalk burley tobacco is
shown in Fig. 8.16 (Long & Weybrew, 1981).

Fig. 8.16
Changes in solanesol levels of burley tobacco during
air-curing (adapted from Long & Weybrew, 1981).
It should be noted that solanesol is present as both the free alcohol and
in bound forms primarily as acetate, palmitate and linolate esters.
Solanesol normally ranges from 1 to 2% in Virginia tobacco and from
1% to as much as 4% in burley tobacco.
The concentration of solanesol in burley tobacco has been shown to
be effected by genetic lines and, more importantly, by production
practices. Water stress, through decreased soil moisture, leads to an
increase in solanesol content as does the length of senescence from
time of topping (Burton, et al., 1989).
Neophytadiene (a C20 isoprenoid polyene) is a tobacco diterpene
whose concentration increases significantly upon curing and aging.
Generally, low levels are present in green tobacco leaf, but
neophytadiene


 Page 278
increases during the yellowing and curing phases. Fig. 8.17 shows the
neophytadiene change in curing burley tobacco (Burton &
Kasperbauer, 1985) and Fig. 8.18 shows the change in flue-curing
Virginia tobacco (Long & Weybrew, 1981).

Fig. 8.17
Changes in neophytadiene content of burley
tobacco during air-curing (adapted from
Burton & Kasperbauer, 1985).

Fig. 8.18
Changes in neophytadiene content during curing
of the flue-cured tobacco.
It has been assumed that neophytadiene is produced by dehydration of
phytol. Phytol is a metabolite from chlorophyll hydrolysis, but it has
been concluded that only 15 to 30% of neophytadiene generated
during flue-curing could be accounted for from this source in the case
of flue-cured tobacco (Amin, 1979). Neophytadiene has been
suggested to be a tobacco flavor enhancer in that it may act as a flavor
carrier by entrapping volatiles in the tobacco smoke aerosol
(Leffingwell & Leffingwell, 1988).
Among other major acyclic isoprenoids are linalool, linalool oxide,
geranylacetone and farnesylacetone. Table 8.9 provides comparative
data on some acyclic isoprenoids, nor-derivatives and neophytadiene
obtained by the steam distillation of various tobacco types (Wilson, et
al., 1982).
Table 8.9 Some major acyclic isoprenoids and
norisoprenoid products isolated from the steam
distillation of tobacco types.
Compound Virginia Burley Oriental
(ppb) (ppb) (ppb)
Linalool 281 160
Linalool oxide 118
Geranyl acetone 181 84 130
Farnesylacetone 16 22 trace
6-Methylhepta-3,5- 40 17
dien-2-one
6-Methylhept-5-en-2- 78 17 78
one

c
Carbocyclic Diterpenoids and Their Degradation Products
Two major classes of diterpenoids are found in tobacco, the
monocyclic cembranoids and the bicyclic labdanoids (Wahlberg &
Enzell, 1987). These diterpenoids are produced in the glandular heads
of the trichomes on the leaf surface as well as in tobacco flowers and
are part of the surface cuticular waxes where they occur along with
the sucrose tetraesters, wax hydrocarbons, esters and alcohols.
Virginia and burley tobaccos contain only the cembranoids, while
Oriental and cigar tobaccos contain both labdanoids and cembranoids.
The variety specific occurrence is due to the fact that Nicotiana
tabacum L. is a hybrid of N. tomentosiformis (which produces
labdanoids) and N. sylvestris (which produces cembranoids). It has
been shown that the biosynthesis of the cembranoids is controlled by
two dominant genes, whereas labdanoid synthesis is controlled by a
single dominant gene (Wahlberg & Enzell, 1987).

 Page 279
d
Cembranoids and Their Degradation Products
More than 50 cembranoids have been isolated from tobacco. The
cembra-2,7,11-triene-4,6-diols are the major cembranoids of tobaccos
(Fig. 8.19) which, for example, have been found in Virginia to be
present at 0.23% by weight (Court, et al., 1993).

Fig. 8.19
Cembranoids in tobacco.
Perhaps more important to the quality of tobacco flavor are the
cembranoid metabolite products produced by oxidative processes and
retro-aldol reactions. More than 60 degradation products of
cembranoids have been isolated from tobacco. Of these degradation
products, some of the major metabolites are shown in Table 8.10.
Table 8.10 Some major cembranoid degradation
products isolated from the steam distillation of
tobacco types.
Compound Virginia Burley Oriental
(ppb) (ppb) (ppb)
Solanone 1725 493 1497
Solanol 545 84 177
Norsolanadione 156 26 137
Solanascone 24 33 trace
Solavetivone 18 15 trace
6-Methylhepta- 101
2,5-dione

Solanone, a major cembranoid metabolite in tobacco, is considered to

be an important tobacco flavor constituent.
e
Labdanoids and Their Degradation Products
(Z)-Abienol is a major labdanoid in the green leaf of Oriental tobacco
and has been shown to be a plant growth regulator. During air-and
sun-curing, the concentration of abienol decreases significantly while
numerous other labdanoids and labdanoid degradation products are
formed. In cured leaf about 20 labdanoids and an additional 25 or
more degradation products are found. (Z)-Abienol has been shown to
undergo various oxidative processes to many of these products
(Wahlberg & Enzell, 1987). Figure 8.20 shows some important
labdanoid derived products in Oriental tobacco.

Fig. 8.20
Some major labdanoid-derived constituents
in Oriental tobacco.
In 1959, norambreinolide (sclareolide), an important constituent of
Oriental tobacco and cigar tobacco, was patented (Schumacher, 1959)
as possessing a cedar character on smoking, although isolation from
tobacco (cigar leaf) was not reported until the early 1970s (Kaneko,
1971). It is now known that norambreinolide and related compounds,
such as sclaral, are responsible for the characteristic cedar-amber
notes of Havana leaf and Oriental tobacco. The important fragrance
compound Ambrox® is found to a much lesser extent in

 Page 280
Oriental tobacco, but this material is a key component in ambergris
derived from whales; the exotic aroma and fixative value of ambergris
is largely due to this material.
The combination of labdanoid-derived flavorants and sucrose
tetraesters present in Oriental tobacco comprises the major flavor-
forming components of this variety (Leffingwell & Leffingwell,
1988). Within the Oriental types, the Greek and Macedonian types
tend to be higher in labdanoid-derived compounds, while the Turkish
Smyrna (Izmir) types are higher in sucrose tetraesters containing the
3-methylvaleric acid component.
Carboxylic Acids
The major carboxylic acids in tobacco are citric, malic, oxalic and
malonic which in total can comprise 14 to 18% in burley and cigar
tobacco, 7 to 10% in Maryland, 6 to 8% in Oriental and 5 to 10% in
Virginia leaf, after curing. A substantial portion of such acids are
complexed as salts with nicotine, ammonia and inorganic anions of
calcium, potassium and sodium. The bar diagram (Fig. 8.21) adapted
from Kalianos (1976) graphically shows the differing acidity profiles
by tobacco type.
Considerable effort has been expended trying to relate such acids to
tobacco quality. Several studies indicate that there is an inverse
relationship to the smoking quality of Virginia tobacco and the
quantity of
 Fig. 8.21
Major organic acids in different tobacco types
(adapted from Kalianos, 1976).
citric and oxalic acids (Kalianos, 1976), although this is probably just
an indicator, and is not due to the absolute amounts of these acids
present in leaf.
The volatile C2-C8 acids are known to be important aroma
compounds in many fruits, foodstuffs and tobacco. Table 8.11 shows
the relative amounts of the major volatile organic acids for various
tobacco types (Kalianos, 1976).
Table 8.11 Volatile organic acids in tobacco.
Acid Virginia Burley Oriental Threshold
(µg/g) (µg/g) (µg/g) (ppb)
Formic acid 597 288 587 45000
Acetic acid 877 372 688 22000
Propionic 13 12 24 20000
acid
Isobutyric 32 29 72 8100
acid
Butyric acid 2 0 0 240
2- 247 26 313 1600
Methylbutyric
acid
Isovaleric 116 20 202 120
acid
2-Butenoic 12 1 3 NA
acid
Valeric acid 1 0 4 3000
3- 4 1 1372 300*
Methylvaleric
acid
Hexanoic 5 3 5 3000
Heptanoic 5 5 6 3000
acid
Octanoic acid 5 12 5 3000
Nonanoic 16 21 24 3000
acid
2-Furoic acid 32 36 125 NA
Benzoic acid 22 14 25 85000
Phenylacetic 36 0 65 10000
acid
* Author's estimate.

It should be noted that the Oriental sample above is an Izmir type

which should have a high 3-methylvaleric acid content and that the
reported values must include the bound acids released by hydrolysis
of sucrose tetraesters.
One of the ways to assess a compound's odor or flavor potency is to
evaluate its minimum detectable threshold level. In the table above are
included the detection thresholds (Leffingwell & Leffingwell, 1991)
for many of the tobacco acids. If one divides the relative concentration
of a material by the threshold one can approximate the relative odor
potency. For example, for acetic acid, 688/2200 = 0.03, while for 3-
methylvaleric acid, 1372/300 = 4.57, indicating the aroma
contribution of the latter would be about 152 × that of the acetic acid.
Table 8.12 shows the concentrations of free acids in an Oriental
tobacco blend and that of the correspond-


 Page 281
Table 8.12 Increase in aromatic acids in
Oriental tobacco smoke due to thermolysis
of sucrose tetraesters.
Acid Tobacco Smoke Change
(µg/g) (µg/g) (%)
Propionic 17.0 66.8 +293
acid
Butyric acid 2.5 9.7 +288
Valeric acid 4.3 3.6 -16
2- 2.1 12.4 +490
Methylbutyric
acid
Isovaleric 5.1 19.7 +286
acid
3- 12.0 62.0 +417
Methylvaleric
acid
Hexanoic acid 7.5 3.9 -48
Octanoic acid 1.7 6.2 +265
Phenylacetic 48.9 19.9 -59
acid
Source: Heckman, et al. (1981).

ing cigarette smoke (Heckman, et al., 1981). This provides an

indication of the increase in free acids due to thermolysis of the
sucrose tetraesters (Leffingwell & Leffingwell, 1988).
Without question, the volatile tobacco acids are some of the most
important contributors to smoke quality and the powerful contribution
from Oriental means that additions to cigarette blends at even low
percentages can have a profound effect on product acceptability.
The higher fatty acids (myristic, palmitic, stearic, oleic, linoleic and
linolenic) comprise about 0.75 to 1.1% in Virginia tobacco and about
0.5% in burley, with palmitic being about 25% of these total acids.
Table 8.13 lists the average amounts of fatty acids found in a 2-year
study on Virginia tobacco grown at Delphi, Ontario, Canada (Court, et
al., 1993).
Table 8.13 Fatty acids in Virginia
tobacco.
Acid %
Myristic acid 0.02
Palmitic acid 0.28
Stearic acid 0.05
Oleic acid 0.11
Linoleic acid 0.18
Linolenic acid 0.35

Tobacco Phenolics
a
Polyphenols
The amount of total phenols varies considerably by tobacco type
suggesting considerable genetic and cul-
Table 8.14 Polyphenols by
tobacco type.
Tobacco 1980 1981
(%) (%)
Virginia 3.13 3.75
Burley 1.78 2.05
Cigar 2.13 2.65
wrapper
Cigar filler 2.03 3.40
Cigar binder 2.27 3.54
Maryland 2.09 3.25
Dark fire- 2.78 3.64
cured
Oriental 1.83 2.09
Source: Smeeton (1987).

tural variability. In a 2-year study the average polyphenol amounts

found were as listed in Table 8.14 (Smeeton, 1987).
Major polyphenolics found in tobacco include chlorogenic acid, rutin,
scopoletin and scopolen, along with materials such as quercetin and
kaempferol. The biogenesis of plant phenolics with relationship to the
pathways relevant to the major groups in tobacco for compounds
formed via shikamic acid has been reviewed by Enzell & Wahlberg
(1980). The amounts of polyphenols for Virginia NC 95 and Burley
21 have been reported (Table 8.15) (Sheen, et al., 1979).
Table 8.15 Typical polyphenols in
tobacco.
Virginia Burley
(mg/g) (mg/g)
Chlorogenic 34.71 12.83
acids
Rutin 7.95 4.00
Scopoletin 0.13 0.06
Scopolin 0.94 0.35
Source: Sheen, et al. (1979).

The chlorogenic acids are primarily 3-O-caffeoylquinic acid with

lesser amounts of the 5-and 6-linked isomers. Scopolin and scopoletin
are the major coumarin compounds in tobacco.
b
Lignin
Lignin comprises as much as 4 to 5% of tobacco weight and is the
most abundant natural organic aromatic polymer found in the vascular
plant kingdom. With cellulose and hemicellulose, lignin is one of the
major cell wall components in leaf. Lignin is composed of a chain of
polymeric phenolic constituents, such as

 Page 282
coniferyl alcohol, p-coumaryl- and synapyl alcohol. In many reviews
on tobacco, lignin has been classified with 'carbohydrates' due to its
association with, and being bound to hemicellulose. In woody plants,
lignin acts as a cementing agent to help bind the matrix of cellulose
fibers (Goheen & Hoyt, 1985).
Lignin is known to produce high levels of phenolic constituents on
thermolysis, such as 2-propylphenol, vanillin, eugenol, guaiacol,
pryrogallol and cresols, which are found in tobacco smoke.
c
Other Phenolics
Many more phenolics are found in tobacco smoke than in leaf, but the
major isolated leaf phenols are shown in Table 8.16 (Wilson, et al.,
1982), albeit they are present in low quantities in leaf.
Table 8.16 Major phenolics isolated from
steam distillation of tobacco types (parts per
billion in leaf').
VirginiaBurleyOriental
Phenol 16 13 39
o-Cresol 9 4 11
m-Cresol 26 8 46
p-Cresol 31 4 18
Guaiacol 80 23 28
Dimethylphenols 9 3 15
4-Vinylphenol 54 2 161
2-Acetyl-3- 2 2
methylphenol
Trimethylphenols 5 4
4-Ethylguaiacol 1 8
4-t-Butylphenol 1 1 8
Eugenol 0.5
4-Vinylguaiacol 14 3 8
4-Allyl-2,6- 52 0.3 2
dimethylphenol
Source: wilson, et al. (1982).

In addition, the vanilla-like phenolic, vanillin, has been isolated from

the steam distillation of a tobacco blend (Green, et al., 1980). It is
probable that some of the phenolics isolated from tobacco by steam
distillation may arise from lignin hydrolysis and, in fact, may be
artifacts.
Phenolics are powerful aroma compounds and are considered to
contribute significantly to smoke quality.
Tobacco Sterols
Sterols, steryl esters and esterified steryl glucosides comprise about
0.1 to 0.3% of tobacco weight. The distribution of sterols for both
green and cured leaf has been studied and they can vary greatly within
the tobacco plant. The major sterols are cholesterol, campesterol,
stigmasterol and b-sitosterol. Table 8.17 provides the amounts of total
individual sterols (after hydrolysis) reported in a sample of Virginia
tobacco (Ellington, et al., 1977) and in two lines of burley tobacco
(Davis, 1976).
Table 8.17 Major sterols in tobacco.
Burley lines
Virginia L-1 L-7
(mg/g) (mg/g) (mg/g)
Cholesterol 0.30 0.18 0.21
Campesterol 0.53 0.23 0.33
Stigmasterol 0.75 0.72 0.82
Sitosterol 0.88 0.38 0.94
Total sterols 2.46 1.51 2.30
SourceL Davis (1976).

Flue-cured, burley and Maryland tobaccos have been reported to have

higher sterol contents than Oriental, fire-cured and cigar leaf tobaccos
(Johnstone & Plimmer, 1959).
Sterols in tobacco are considered to be a potential health negative in
that the parent polycyclic structure may form polynuclear
hydrocarbons on pyrolysis (e.g. stigmasterol forms benzo(a)pyrene at
750ºC) (Johnstone & Plimmer, 1959; Stedman, 1968).
Inorganics
Significant amounts of calcium, potassium, magnesium, chlorine,
phosphorus and sulfur are found in the ash of tobacco (as well as
sodium), as shown in Table 8.1. In addition, both free and complexed
ammonia and nitrate are found in tobacco. The topic of ammonia in
tobacco has previously been discussed in this section.
The tobacco content of these inorganics is in large part due to soil and
fertilizer practices. Lower amounts of many other inorganic anions
(e.g. zinc, aluminum titanium, strontium, silicon, rubidium, nickel,
manganese, lithium, lead, iron, copper, chromium, cesium, boron,
barium and arsenic) have been found in tobacco (Johnstone &
Plimmer, 1959; Stedman, 1968). The presence of the major metallic
anions certainly impacts (for example, magnesium and potassium
accelerate) the burning characteristics of tobacco leaf and the ash-

 Page 283
holding properties of cigarettes. Phosphorous and chloride tend to
slow the burning process. In fact, high chloride levels provide both
negative burning and taste characteristics.
Tobaccos with high nitrate content are assisted in the burning process
as nitrate is itself a combustible material. In air-cured tobaccos, the
nitrate tends to accumulate in the stem portion and is considered a
major reason why the smoking properties of burley stems are inferior
to Virginia stems. For many years, major cigarette manufacturers
'washed' their burley stems to reduce the nitrate content. This served
two purposes: improved the smoking properties and removed the
nitrate oxidant which lowered the level of nitrosamines in smoke.
The trace elements (such as iron) in tobacco can have a profound
effect on tobacco quality. For example, iron has been implicated in the
speckling effects that develop in the so-called 'grey tobacco'
deficiency of Virginia tobacco (Tso, 1972; Tso, 1990).
Summary
Tobacco is a Complex plant material from a chemical composition
standpoint. No other plant material has been studied more extensively
in the history of man. Yet even today, the quality of tobacco is judged
largely by empirical experience and subjective sensorial evaluation. It
is not possible to replicate the smoking properties of the tobacco leaf
synthetically. Yet, through chemical knowledge and genetic advances,
there is now a much better understanding of tobacco growth and
important constituents that allows for improvements in quality using
the best laboratory in the world through optimization of the biology
of tobacco itself in the tobacco growing field.
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8B
Alkaloid Biosynthesis
L.P. Bush
University of Kentucky
Lexington, Kentucky, USA
Tobacco Alkaloids
Nicotine is the principal alkaloid in Nicotiana tabacum. Of the many
species of Nicotiana, 50 to 60% have nicotine as the predominant
alkaloid and, based on the amounts of alkaloid accumulation in the
leaves, nicotine, nornicotine, anatabine and anabasine are the major
alkaloids present. Nornicotine is the principal alkaloid in 30 to 400%
of the Nicotiana species and anabasine is the principal alkaloid in four
species. Anatabine is not usually the principal alkaloid in any species,
but does accumulate to relatively higher amounts in three species.
Many other pyridyl bases plus many derivatives of nornicotine,
anatabine and anabasine have been reported to be present in tobacco
(Tso, 1990; Bush, et al., 1993). Most of these so-called minor
alkaloids are present in less than 50 µg/g (dry weight basis) and many
others are present in nanogram amounts. Many of these minor
alkaloids are probably the result of aberrant metabolism or catabolic
products of the major alkaloids.
Nicotine and nornicotine have received the most research attention
because of their importance in tobacco quality. Nicotine content seems
to be very important as a positive attribute of quality, whereas
nornicotine content has a negative impact on tobacco quality.
Consequently, this review will focus mainly on these two alkaloids
and their derivatives. Species with nicotine as the principal alkaloid
tend to have higher levels of alkaloid than other species, and the
principal route of nornicotine formation is by nicotine demethylation.
Nicotine and nornicotine levels in tobacco are affected by genetics,
environmental conditions and cultural practices.
Many reports have been published detailing the effects of these
parameters on nicotine and nornicotine accumulation in tobacco
leaves, but little is known about the specifics of how these parameters
cause change in alkaloid accumulation. The gross action of these
parameters has been covered in many earlier reviews and the reader is
referred to these works ( Bush & Saunders, 1977; Chaplin & Miner,
1980; Bush & Crowe, 1989; Tso, 1990). The general conclusion from
studies on environmental conditions and cultural practices is that
anything that improves the growth of the plant will increase alkaloid
formation and accumulation. Increased nitrogen fertilizer, decreased
topping height, increased length of photoperiod, decreased soil
temperature, increased time between topping and harvest and
increased sucker control all increased nicotine accumulation and were
most likely the result of increased leaf yield (Fig. 8.22).

Fig. 8.22
The influence of soil nitrogen fertilization on dry matter yield and
alkaloid content.
Flue-cured tobacco is grown under lower nitrogen fertility levels than
burley tobacco, and in flue-cured tobacco there is a positive
correlation between leaf nitrogen and nicotine concentration (Woltz, et
al., 1948). However, in burley tobacco, tissue nitrogen and total
alkaloids may not be correlated (Bortner, et al., 1960) However,
increased nitrogen fertility does increase total alkaloids (Fig. 8.22)
(Atkinson, et al., 1969; Tso, 1990). Nitrogen utilized for nicotine
biosynthesis may be absorbed before or after topping (Yoshida &
Takahashi, 1961). However, increased dry weight is concomitant with
nitrogen uptake in burley tobacco (Atkinson, et al., 1977) whereas in
flue-cured tobacco nitrogen uptake ceases before the surge in

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alkaloid biosynthesis associated with topping (Raper & McCants,
1966). As topping height decreased, nicotine concentration in the
remaining leaves at harvest increased (Chaplin, et al., 1964). In these
experiments leaf dry weight at harvest decreased slightly because of
the fewer leaves, but alkaloid concentration and total alkaloid
accumulation in the leaves increased. Total alkaloid level was greatest
in the upper leaves and decreased in leaves from descending stalk
positions (Bowman, et al., 1958).
The relative change in nicotine content in leaves from bottom to top of
the stalk may be 200 to 300%. Also, as the number of days between
topping and harvest increase, nicotine content of the plant increases.
However, there is an interaction with leaf stalk position and time after
topping (Fig. 8.23). Leaves from the lower stalk position have
increased nicotine during the first 10 to 14 days after topping, but as
leaf maturation and senescence continue, nicotine levels decrease to
levels equal to or less than the initial level. Nicotine concentrations in
leaves from the middle and upper stalk positions continue to increase
for several weeks after topping, and this increase was greater in the
upper leaves than the middle leaves (Bowman, et al., 1958). The
change in nicotine concentration between topping and harvest may be
400%. During the time between topping and harvest, leaf dry weight
also is increasing, but nicotine concentration is increasing at a faster
rate and total nicotine produced per land area accumulates at even a
greater rate. Greater sucker control after topping also increases
nicotine level in leaves. Because sucker control chemicals may alter
other plant processes, this phenomenon is best measured in hand-
 Fig. 8.23
Nicotine accumulation in leaves from different stalk positions with time after
topping. (D.O.H. is date of harvest.)
suckered experiments. Relative leaf nicotine concentrations were
found to be 1.00, 1.28, 1.48 and 1.57 from plants hand suckered 0, 1,
2 or 3 times. The increased nicotine biosynthesis is mainly the result
of enhanced root growth, which is the site of nicotine synthesis, and
the stimulation by removal of the auxin source (apical meristem). It
must be emphasized that there are often interactions among these
environmental and cultural parameters and the resulting alkaloid
accumulation is affected mainly by dry matter accumulation relative
to alkaloid formation and accumulation.
Genetic Control of Syntheses
Genetic control of alkaloid accumulation in tobacco has been
described for nicotine and nornicotine. These two systems are
different, and an understanding of their mechanism of regulation is
very important to the success of molecular biology programs aimed at
altering the alkaloid content of leaf product. The level of nicotine in
tobacco is controlled by two nonlinked loci referred to as Nicl and
Nic2 or A and B (Legg, et al., 1969). The mutant alleles are recessive
for lowered nicotine accumulation. Commercial cultivars are
considered homozygous dominant at the two loci. The relative dosage
effect of the homozygotes at locus Nicl was 2.4 times greater than the
homozygotes at locus Nic2 (Legg & Collins, 1971).
Nornicotine formation is primarily the result of demethylation of
nicotine to nornicotine (Dawson, 1952). Also, two potential loci for
expression of demethylation occur in N. tabacum, one from each of
the progenitor species N. tomentosiformis and N. sylvestris (Mann, et
al., 1964). The primary site of nicotine demethylation to nornicotine
occurs in the leaf, however the enzyme is found in all tissues. The
gene from N. tomentosiformis causes demethylation prior to curing
while the gene from N. sylvestris causes demethylation during curing
(Wernsman & Matzinger, 1968). The gene from N. tomentosiformis
seems to be the active allele in commercial tobaccos (Griffith, et al.,
1955; Burk & Jeffrey, 1958; Mann, et al., 1964). Only one dominant
allele at either of the loci is necessary for demethylation to occur, and
there is a very high mutation rate at this locus for nicotine
demethylation during senescence. Because of the negative impact that
nornicotine has on smoke quality, it is this high mutation rate that
makes it necessary for plant breeders to continually monitor
genotypes for nornicotine accumulation.

 Page 287
Plant Distribution
Alkaloid accumulation within plant parts is vastly different. Leaf
lamina generally contains the greatest concentration of alkaloids with
decreasing amounts in descending order found in the roots, midribs,
stalks, flowers and seed (Bush & Crowe, 1989). Root tips, the site of
nicotine synthesis, often contain greater concentrations of alkaloids
than leaf lamina, but if the whole root system is included in the
sample, dilution occurs and the alkaloid concentration decreases.
Alkaloid concentration in midribs and stalks is about 20% and 12%,
respectively, of that in the lamina. Flowers contain about 10% of the
alkaloid concentration of the upper leaves at the time of flowering.
Seeds contain very low concentrations of free alkaloids, but extraction
with a strong acid solvent released 0.37 mg alkaloid per gram seed dry
weight (Weeks & Bush, 1974).
Most importantly for tobacco utilization is the distribution of alkaloids
within the leaf. Within the midrib the alkaloid concentration decreased
from the tip to the base of the leaf, with the greatest decrease
occurring 25 to 30% of the distance from the leaf tip (Burton, et al.,
1992). The lateral veins had greater alkaloid levels than the mid-rib
and also decreased from the tip to the base of the leaf and from the
leaf margin to the mid-rib. The accumulation of alkaloids in the
lamina increased from the center to the margins (Fig. 8.24)
(Andreadis, et al., 1939; Burton, et al., 1992). The alkaloid
concentration is much greater at the margins nearer the base of the
leaf than elsewhere. Within the leaf, dis-
 Fig. 8.24
Distribution of nicotine in cured leaf; x-axis: 0,0
coordinates are the tip of the leaf with the mid-rib
samples on the line from coordinates 0,0 to 0,80.
tribution of nicotine and the minor alkaloids are similar. The best
portion of the leaf to sample to obtain an average value for the whole
leaf is a 3 to 6 cm transverse segment from about a third of the
distance down from the tip of the leaf (Gay & Bush, 1986).
Nicotine Synthesis
Nicotine was first described in 1809, isolated and purified in 1828 and
the structure and synthesis determined in the late 1800s (review by
Jackson, 1941). Since that time many reports have appeared in the
literature detailing the synthesis of nicotine and related alkaloids.
Many enzymes that are involved in the biosynthetic pathway have
been named, isolated from tobacco roots and partially characterized.
However, reproducible synthesis of nicotine in a cell-free system has
not been reported. Nicotine is synthesized mainly in the roots and
most investigations have used root tip tissue to elucidate the enzymes
and biosynthetic pathway.
Carbon dioxide is readily incorporated into nicotine, but the principal
intermediates are a 3-carbon sugar derivative and aspartic acid for the
pyridine ring (Yang, et al., 1965) and putrescine for the pyrrolidine
ring (Jackanioz & Byerrum, 1966). The tobacco alkaloid biosynthetic
pathways are summarized in Fig. 8.25. The phosphoglyceraldehyde
probably comes directly from carbon fixation of photosynthesis as
Robinson (1974) reported that during rapid alkaloid biosynthesis 17%
of the carbon fixed was found in nicotine. The aspartic acid most
likely is an early transamination product of malic acid. These two
substrates, aspartic acid and a 3-carbon sugar or the reaction product
of these two substrates, are probably translocated to the root for
further processing to yield nicotine.
Mizusaki, et al. (1973) reported the activities of ornithine
decarboxylase (ODC), putrescine N-methyltransferase (PMT) and
methylputrescine oxidase (MPO) to be much higher in tobacco roots
than leaves and the activity increased in the roots after topping.
Saunders and Bush (1979) reported PMT and MPO increased in
normal alkaloid genotypes after topping, but that MPO did not
increase following topping in a low alkaloid genotype. PMT activity
in the roots was proportional to leaf nicotine in several related and
unrelated high and low alkaloid genotypes (Yoshida, 1973; Saunders
& Bush, 1979). Pudliner (1980) showed that ODC increased in both
normal and low alkaloid genotypes as well as citrate synthase, an
indicator of metabolic activity. These results indicate that the N-

 Page 288

Fig. 8.25
Pathway for alkaloid biosynthesis in tobacco.
1 = quinolinic acid phosphoribosyltransferase; 2 = nicotinic acid mononucleotide
glycohydrolase; 3 = arginine decarboxylase; 4 = ornithine decarboxylase; 5 =
putrescine methyltransferase; 6 - methylputrescine oxidase; 7 = nicotine
demethylase.
methyl-D'-pyrrolinium is formed in the root and that PMT is the most
limiting enzyme in the pathway.
Consequently, the role of putrescine supply as well as utilization may
be a regulatory step in synthesis of nicotine, and the isolation of the
gene for PMT in tobacco has confirmed this conclusion (Hibi, et al.,
1994). The amount of nicotine formed was associated with the
availability of putrescine (Bush, et al., 1984; Miller, et al., 1983).
Putrescine has a central role in several biosynthetic pathways (Fig.
8.26) All the products of putrescine utilization shown in Fig. 8.26
accumulate in tobacco tissues at some time during the life cycle.
Cinnamoyl putrescine accumulation in flowers, bud leaves and cell
lines has been found as high as 5% to 9% of dry weight (Berlin &
Witte, 1982; Snook, et al., 1986; Snook, et al., 1987). The polyamines
accumulate in low alkaloid genotypes to a much greater extent than in
normal genotypes, with spermidine being the predominant polyamine
present

Fig. 8.26
Involvement of putrescine in metabolic pathways of tobacco. From Bush, et al.,
1993, published with permission of Chapman & Hall.
(Madsen, et al., 1984). g-Aminobutyric acid (GABA) is a major
component of amino acid chromatograms of tobacco and conversion
of putrescine to GABA has been demonstrated. Two of the pathways
for utilization of putrescine, methylation for nicotine biosynthesis and
aminopropylation for polyamine formation, have the same substrate.
Polyamine levels are elevated during rapid cell proliferation, such as
occurs in tumor growth (Bagni, et al., 1972; Russell, 1973). In
tobacco callus, high growth rates are usually associated with low
nicotine accumulation (Takahashi & Yamada, 1973; Miller, et al.,
1983).
Putrescine levels are normally low in tobacco, but Yoshida (1967)
reported elevated levels with deficiencies of boron, potassium and
sulfur. Increased levels of putrescine should be associated with tissues
that contain high ODC activity, and Purlliner (1980) found tobacco
callus has 10-times higher activity than roots of topped plants. The
low alkaloid levels but high ODC activity in tobacco callus suggest
that putrescine formation is not limiting nicotine biosynthesis, but
perhaps utilization of putrescine and S-adeno-sylmethionine for
polyamine biosynthesis is limiting the availability of putrescine for
nicotine biosynthesis. Specific inhibitors of ODC and arginine
decarboxylase activity did lower nicotine accumulation in tobacco
callus (Philips, et al., 1992).
The next step in nicotine biosynthesis, oxidation of methylputrescine,
raises many intriguing questions, N-methylputrescine oxidase does not
have great in vitro substrate specificity. Mizusaki, et al. (1972)
demonstrated that MPO utilized putrescine and cadaverine 40% as
readily as methylputrescine. Enzyme preparations from a nicotine
accumulator and a nornicotine accumulator (a nicotine to nornicotine
converter) oxi-

 Page 289
dized putrescine and methylputrescine equally (Saunder & Bush,
1979). Since oxidation of putrescine to 4-aminobutanal could lead to
formation of nornicotine, some explanation is needed to account for
the disproportionate differences in nicotine and nornicotine
accumulation. Several hypotheses may be suggested, but
compartmentalization of the reactions or very rapid dehydrogenation
of 4-aminobutanal to 4-aminobutyric acid seems most likely. These
results suggest that substrate specificity of MPO is not a major factor
in nicotine or nornicotine accumulation in Nicotiana species.
However, MPO may be regulated in concert with PMT and thus may
be a secondary control of nicotine biosynthesis.
Availability of nicotinic acid for the pyridine moiety of nicotine is an
important regulatory point for nicotine biosynthesis. Quinolinic acid is
an efficient precursor of the pyridine ring (Yang, et al., 1965) (Fig.
8.27). Mann and Byerrum (1974) were the first to report that
quinolinic acid phosphoribosyltransferase (QPT) activity was very
high in roots but not leaves of tobacco and suggested that this enzyme
was important in nicotine biosynthesis. Subsequently, Saunders and
Bush (1979) found nicotine levels in leaf tissue to be proportional to
Q PT activity in roots of different alkaloid genotypes. Enzyme activity
increased with treatment to induce nicotine accumulation in all
genotypes except the homozygous recessive low alkaloid genotypes.
The increased Q PT activity was apparently unique to nicotine
accumulating tissues, as citrate synthase, a metabolic indicator
enzyme, activity did not differ between alkaloid induction treatments
or genotypes (Pudliner, 1980).
It is generally accepted that QPT is the principal regulatory enzyme
for nicotinic acid synthesis for nicotine biosynthesis. Other enzymes
in the pyridine nucleotide cycle have secondary regulatory control of
 Fig. 8.27
Pyridine nucleotide cycle and nicotinic acid availability for nicotine
biosynthesis. From Bush et al., 1993, published with permission of
Chapman & Hall.
nicotinic acid as represented by width of arrows in Fig. 8.27. The Km
(Michaelis-Menten) for nicotinic acid mononucleotide glycohydrolase
is high and probably prevents depletion of the nicotinamide
mononucleotide pool for nicotine biosynthesis to ensure continued
nicotinic acid adenine dinucleotide availability for cellular activity
(Wagner, et al., 1986a; Wagner, et al., 1986b). Nicotinic acid
mononucleotide glycohydrolase did not have an apparent regulatory
purpose in anabasine biosynthesis in N. glauca (Fannin & Bush,
1988).
The general conclusion of the biochemistry and physiology is that
PMT and QPT are the most rate limiting enzymes in biosynthesis of
the two immediate precursors for nicotine. Other substrates or
enzymes in the pathways may have a regulatory function in certain
situations. In plants the significance of these findings is somewhat
diminished by experiments with replacement of enzyme product and
the resultant lack of normal nicotine accumulation. Purification of
QPT and development of transgenic plants with altered QPT activity
and nicotine accumulation, and better knowledge of nicotine synthase
in vivo or in vitro appears to be required before unequivocal
conclusions are made.
Normal alkaloid tobacco genotypes (Nic1Nic1Nic2Nic2) do not
accumulate more nicotine with addition of N-methylputrescine and
nicotinic acid as observed in the low alkaloid genotypes. However,
even in these substrate addition studies, low alkaloid genotypes do not
accumulate nicotine equal to the normal genotype controls. These
results suggest that nicotine synthase may be rate limiting for nicotine
accumulation in some instances. Studies on nicotine synthase have
been limited primarily to aberrant alkaloid synthesis as isolation of the
enzyme in vitro has proven very difficult. Nicotine synthase activity
in a cell-free system was reported by Friesen and Leete (1990), but
their results have not been repeated by other researchers. Significant
in their report of the enzyme was the absence of activity during the
first 100 hours of seed germination. This observation is consistent
with the in vivo report of Weeks and Bush (1974) that nicotine does
not begin to accumulate until about 96 hours after initiation of
germination. Certainly, additional work on in vivo and in vitro activity
of nicotine synthase must be completed before the entire regulatory
control of nicotine biosynthesis is understood.
Other Alkaloid Syntheses
Anabasine and anatabine are formed by similar decarboxylation
condensation reactions as nicotine and it is

 Page 290
not known if these reactions are catalyzed by the same enzyme (Fig.
8.25). Anatabine biosynthesis seems to be entirely dependent upon
QPT, the pyridine nucleotide cycle and a synthase. Certainly, the
recognition of the nicotinic acid and the 1,2-dihydropyridine most
likely would be the same as for nicotine synthase. The similarity of
condensation reaction with the N-methyl pyrrolinium or the 2,5-
dihydropyridine in nicotine or anatabine biosynthesis is problematic.
Anabasine biosynthesis may be even more similar to nicotine as the
1,2-dihydropyridine would react with the D'-piperideine (Fig. 8.25).
The last of the four major alkaloids of tobacco, nornicotine, is
apparently formed mainly by demethylation of nicotine. Nicotine
demethylation is nearly irreversible (Dawson, 1951; Leete, 1984)
(Fig. 8.25). There are many reports in the literature on the physiology
of the demethylase enzyme but very few on in vitro properties.
Demethylation of nicotine occurs in both growing and senescing
tobacco tissues. As noted in the beginning, many Nicotiana species
accumulate nornicotine as the major alkaloid, indicating that nicotine
was first formed and demethylated readily prior to sequestering of the
terminal alkaloid. In many of these plants the demethylation obviously
occurs during plant growth; however, in some plants and in N.
tabacum converter genotypes the demethylation occurs during leaf
senescence. Many tissues contain nicotine demethylase, but the
enzyme is only expressed in the presence of nicotine. Pith tissue of N.
otophora, a low alkaloid accumulating plant, is a good source of
extractable nicotine demethylase and is used for reports on the
characterization of the enzyme (Chelvarajan, et al., 1993).
Inactivation of nicotine demethylase in N. tabacum may be useful to
alter the alkaloid quality of commercial leaf.
Summary
Alkaloids in tobacco are extremely important because they are an
essential factor in leaf quality and provide a physiological stimulus
which makes the use of tobacco products pleasurable. Alkaloids in
Nicotiana species have been ascribed many functions and they appear
to be important in the evolution of the genus. However, as important
as the alkaloids are in utilization of tobacco, growth and development
of the plant will proceed normally in the absence of accumulation of
alkaloids. Despite the many reports on research with tobacco alkaloids
we do not understand the function in the plant nor do we completely
understand the regulation of biosynthesis and accumulation in the
plant.
References
Andreadis, T.B., Toole, E.J., Binopoulos, X. & Tsiropoulos, J. (1939)
Distribution of nicotine in the tobacco plant. Z. Untersuch. Lebensm.,
77, 26272.
Atkinson, W.O., Bush, L.P. & Sims, J.L. (1977) Dry matter and
nutrient accumulation in burley tobacco. Tob. Sci., 21, 812.
Atkinson, W.O., Ragland, J.L., Sims, J.L. & Bloomfield, B.J. (1969)
Nitrogen composition of burley tobacco. I. The influence of irrigation
on the response of burley tobacco to nitrogen fertilization. Tob. Sci.,
13, 1236.
Bagni, N., Serafini Fracassini, D. & Corsini, E. (1972) Tumors of
Scorzonera hispanica. Content in polyamines. Z. Pflanzenphysiol. 67,
1923.
Berlin, J. & Witte, L. (1982) Metabolism of phenylalanine and
cinnamic acid in tobacco cell lines with high and low yields of
cinnamoyl putrescines. J. Natl Prod., 45, 8893.
Bortner, C.E., Wallace, A.M. & Hamilton, J.L. (1960) Differences in
potassium, nitrogen and total alkaloid concentration of ten burley
tobacco varieties. Tob. Sci.., 4, 1515.
Bowman, D.R., Nichols, B.C. & Jeffrey, R.N. (1958) Time of
harvest its effect on the chemical composition of individual leaves of
burley tobacco. Tenn. Agric. Exp. Sta. Bull., 291.
Burk, L.G. & Jeffrey, R.N. (1958) A study of the inheritance of
alkaloid quality in tobacco. Tob. Sci., 2, 13941.
Burton, H.R., Dye, N.K. & Bush, L.P. (1992) Distribution of tobacco
constituents in tobacco leaf tissue. 1. Tobacco-specific nitrosamines,
nitrate, nitrite and alkaloids. J. Agric. Food Chem., 40, 105055.
Bush, L.P. & Crowe, M.W. (1989) Nicotiana alkaloids. In: Toxicants
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8C
Leaf Surface Chemistry
G. Wagner
University of Kentucky
Lexington, Kentucky, USA
Introduction
Any review of tobacco surface chemistry written in the 1990s should
pay tribute to the contributions of Ray F. Severson, who passed away
in 1993. So many who have been involved with this research area and
the impact of tobacco surface chemicals on pest interactions, flavor
and aroma, etc. have exploited (and indeed relied upon) the huge
catalog of excellent information and technical advances generated by
Severson and his colleagues. His excellent science, energy,
organizational talents, his devotion to the tobacco system and his
friendship are missed.
It has been said that a genuinely successful scientist is one who has
generated new and significant information that not only stands the test
of time but stimulates others to build upon a discipline, and perhaps
beyond. Ray achieved all of this in his too short career. This section is
dedicated to the memory and accomplishments of Ray F. Severson.
Were he alive, he would rightfully have written this section.
This review will attempt to present the state of our understanding of
the chemistry, modes and site of biosynthesis, stability and regions of
deposition of the main surface constituents of tobacco, the diterpenes,
sugar esters, surface waxes and volatiles. Methods for their isolation
and characterization will be discussed only briefly with references
cited. The relative amounts of constituents on green tobacco will be
considered briefly. Discussion of smoke chemistry as it relates to
surface chemistry of the green plant will be left entirely to the section
following this one, and discussion of surface chemical, pest/disease
interactions will be limited, as a section dealing with this subject is
included in this monograph.
The Nomenclature of Tobacco Surface Chemicals
Various general terms have been used to describe surface components
in the tobacco system. These include surface chemicals, leaf surface
chemicals, surface lipids, cuticular components, cuticular and
epicuticular lipids, exudate chemicals or constituents, surface waxes,
etc., and various versions of these. Understanding of the chemistry,
biosynthesis, sites of synthesis and deposition of tobacco surface
components has progressed to the point that some of these terms are
misleading and incorrect. For example, 'cuticular' implies close
association with the cuticle, which is a distinct surface component
having very distinct chemical composition. Tobacco trichome exudate
components such as common diterpenes and sugar esters are largely
not associated with the cuticle but are secreted into a space under it
and outside the gland cells producing them. Subsequent to their
secretion, components generally, in part, escape the cuticle barrier and
flow to the surface above the cuticle. Perhaps due to their amphipathic
nature, they may partially impregnate the cuticle. So, 'epicuticular' (on
top of the cuticle), 'cuticular', and 'subcuticular' may all be partially
correct, but no one term is solely correct. Yet, exudate components are
often referred to as 'cuticular' even though they are more important to
tobacco production and use than the cuticle. Further, the surface
components most important to tobacco production (exudate diterpenes
and sugar esters) are not formed from lipid metabolism. Research
shows that acyl groups of sugar esters are not biosynthesized via fatty
acid synthase related reactions that form membrane and storage lipids
in all living organisms but are produced by a newly discovered, novel
pathway (termed a-KAE, see below) that is unrelated to lipid
metabolism (Kroumova, et al., 1994).
So, what general term is a reasonable one? It is suggested that the
term 'surface chemistry' be used. Surface biochemistry would be more
correct and allow one to distinguish plant-produced from
exogenously-applied (i.e. pesticide residues) chemicals. But, tobacco
is a business in which chemistry is central, so surface chemistry is
perhaps better in this case. Misunderstanding of this general term
could only come

 Page 293
if one nit-picks about the extent of contact with the atmosphere that
the word 'surface' implies (i.e., inner cuticle may not contact the
atmosphere). Surface chemistry is more appropriate than leaf
chemistry since, for example, stems, mid-ribs, etc. often contain
higher levels of the same surface chemicals than leaves, and these
tissues are important to tobacco production and use. Surface chemistry
is also a correct, general term for cured and processed leaf.
Subdivision of the main tobacco surface chemicals might be into the
three main groups, the trichome exudate, diterpenes, sugar esters, and
the surface waxes (includes all cuticle components, see below). Minor
groups are the volatiles and miscellaneous components (see below).
Fig. 8.28 suggests a perhaps improved nomenclature scheme for
tobacco surface chemicals of green tobacco.

Fig. 8.28
Proposed nomenclature for tobacco surface chemicals
(based on our current understanding of their
chemistry, biochemistry, tissue location, etc.).
In addition, the many terms for tobacco exudate diterpenes that have
emerged since their discovery (i.e., thunbergol, 4-epiisocembrol,
duvanes, duvatrienes, etc.) should yield to more standard chemical
nomenclature and use. This means adoption of the cembrenoid and
labdenoid nomenclature (Wahlberg & Eklund, 1992). Also, since a
significant number of tobaccos contain sucrose esters and/or glucose
esters, 'sugar esters' is perhaps a better general term for these
important components. Glycolipids is inappropriate, again because
sugar esters are not derived from lipid metabolism.
Tobacco Surface Chemistry: General Aspects
The surface chemistry of tobacco and methods for constituent
characterization were generally reviewed by Severson, et al. (1985a).
The catalog of surface chemical composition (particularly of exudate
components) contained in that review has been extremely helpful in
exploiting the genetic diversity of the many available tobacco lines for
the purposes of studying pest-interactive properties of exudates and
the enzymology and site of their synthesis and deposition. That review
remains the most comprehensive one published.
As defined in Fig. 8.28, the main groups of the green tobacco surface
chemicals are:
(1) the cembrenoid and labdenoid diterpenes,
(2) the sugar esters, and
(3) the surface waxes.
Minor groups are:
(4) volatiles produced at the surface, and
(5) minor components found in certain tobaccos such as N-
hydroxyacylnornicotine in N. stocktonii (Zador & Jones, 1986),
cembrene in N. tabacum (Takagi, et al., 1980), and trichome
associated metals such as 210Po.
Various metals are often found to be enriched in various cell types of
plant trichomes. The significance of this is not understood. Gland cells
of tobacco trichomes often contain very large calcium oxalate crystals
(Vogeli-Lange & Wagner, unpublished data). Commonly occurring
examples of each group are shown in Fig. 8.29. The last group (minor
components) will not be discussed further. Degradation products of
exudate diterpenes and other surface constituents occur in aged or
cured tissue as a result of microbial or chemical action. This aspect is
discussed in the next chapter.
Trichomes are the tissue type most important to tobacco surface
chemistry, at least from a commercial point of view. In addition to
their role in producing exudates, trichomes affect leaf wettability and
interaction of the plant with light (Duffey, 1986; Fahn, 1988; Brewer,
et al., 1991; Wagner, 1991). Various types of trichomes occur on the
aerial surfaces of tobaccos. These may be branched or unbranched
glandless types, generally unbranched glanded types, or hydathodes
(Severson, et al., 1985a). Only glanded types are thought to produce
nonvolatile secretions. Exposure to iodine vapors (to stain terpenes)
stains most but not all TI 1068 glandular trichomes bearing exudate
(unpublished data). Also, this is observed when stain-

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Fig. 8.29
Examples of major tobacco surface chemicals.
ing with rhodamine is used to reveal sucrose esters (SE) (see below).
Therefore, it is possible that not all secreting trichomes on a given
surface are alike in their exudate profiles. Subcellular sites involved in
exudate terpene biosynthesis clearly include the plastids and
endoplasmic reticulum, but details are not known (Wagner, 1991).
Cembrenoid and Labdenoid Diterpenes
The least chemically diverse, but usually the most abundant of the
three major groups of surface chemicals is the first, the cembrenoid
and labdenoid diterpenes. The cembrenoid di-alcohols, a- and b-
2,7,11-cembratriene-4,6-diol (CBT-diol), are more abundant and
widely distributed than their corresponding monools (CBT-ol), a- and
b-2,7,11-cembratriene-4-ol (also known as thunbergol or isocembrol,
and 4-epiisocembrol, respectively). The cembrenoid monools are also
found in conifer oleoresins but have been found in a few other plant
species (Wahlberg & Eklund, 1992). Also, the a and b epimers have
been called a and b duvatrieneols and duvatrienediols. But, as noted
above, these terms should now be discontinued. Absolute
configurations of a-CBT-ol and b-CBT-ol were determined to be
(1S,2E,4R,7E, 11E)- and (1S,2E,4S,7E,11E)-cembra-2,7,11-trien-4-ol,
respectively (Wahlberg & Eklund, 1992). Configuration at carbon 6 of
the diols is R. The a and b epimers generally occur in a ratio of 2:1 to
3:1. This ratio may depend on developmental stage (Marysia, et al.,
1993). Similar ratios of these are formed in cell-free reactions (Guo &
Wagner, 1995a).
Labdenoids are not as common as cembrenoids but are more diverse
in structure and are found in many tobaccos. Figure 8.30 shows some
of the various labdenolds that occur in Nicotiana species. Most
commonly found is cis-abienol. Labdenoid diterpenes are also
reported from Nolane and Juniper species (Garbarino, et al., 1988;
San Feliciano, et al., 1992).

Fig. 8.30
Labdane diterpenes in tobacco trichome exudates.
Cembrenoid and labdenoid diterpenes are generally recovered from
plant surfaces by washing with acetonitrile, methylene chloride,
acetone, chloroform, etc., then separated/identified and quantitated as
their trimethysilyl derivatives using GC or GC-MS (Arrendale, et al.,
1990; Severson, et al., 1985b). Alternatively, separation without
derivatization and with full recovery may be made using HPLC (Guo
& Wagner, 1995b; Guo, et al., 1994). Thin layer Chromatography
(TLC) methods can separate or partially separate these components,
but components are not accurately quantified by this method.
Acetonitrile preferentially removes diterpenes and sugar esters, while
the other less polar solvents also remove hydrocarbons.
The cembrenoid and labdenoid diterpenes are undoubtedly
synthesized in trichome glands alone. This was shown directly for
CBT-ols and -diols, cisabienol, labdenediol and sclareol in various
tobaccos by comparing the ability of isolated glands versus epidermis-
without-glands versus subepidermal tissue to biosynthesize these
compounds (Kandra & Wagner, 1988; Wagner, 1991; Guo, et al.,
1994; Guo & Wagner,

 Page 295
1995a; Guo & Wagner, 1995b). Isolated glands of tobacco
introduction (TI 1068) biosynthesize a and b CBT (and sucrose esters)
in the light from the simple precursors bicarbonate or sucrose (Kandra
& Wagner, 1988). Therefore, glands are capable of forming complex
diterpenes and sugar esters independent of epidermis or subepidermis.
It is possible that, in situ, carbon is supplied to gland cells as sucrose
from the epidermis. But in isolated glands, sucrose is inverted to
glucose and fructose prior to utilization for sucrose ester synthesis
(Kandra & Wagner 1988). So, the initial precursor in situ is probably
glucose (formed in the gland or derived from translocated sucrose, or
both).
Current understanding of the mechanism of cyclization of
geranylgeranyl pyrophosphate to form CBT-ol in tobacco indicates
that cembrene and 6hydroxycembrene are not intermediates and that
the a and b CBT-ols are direct products of cyclization (Guo & Wagner,
1995a). It is presumed that the monols are converted to diols by
subsequent hydroxylation (possibly by a P450 type mechanism), but
this has not been shown. Variation in monol versus diol ratios in
different tobaccos may reflect efficiency of subsequent hydroxylation.
It has been concluded from plant breeding studies that CBT-diol
synthesis is controlled by two dominant traits (Severson, et al.,
1985a). These may encode for CBT-ol synthase and a hydroxylating
enzyme. Similarly, it is suggested that cis-abienol biosynthesis is
directed by a single dominant gene (Severson, et al., 1985a). This
gene may encode cis-abienol synthase. The enzymes and genes for
synthesis of certain related plant diterpenes (i.e. casbene), are better
understood than those for cembrenoids and labdenoids of tobacco
(Mau & West, 1994). However, we recently demonstrated and
partially purified cis-abienol synthase (Guo, et al., 1994), CBT-ol
synthase (Guo & Wagner, 1995a), and sclareol/labdenediol synthase
(Guo & Wagner, 1995b) activities from various tobaccos. These
soluble enzymes cyclize geranylgeranyl pyrophosphate to form
macrocyclic CBT-ols and bicyclic labdenoid-ols. The biochemical
characteristics and apparent mechanism of cyclization of these
enzymes are similar to those of monoterpene and sesquiterpene
cyclases found in other plants (Gershenzon & Croteau, 1993).
Methods used in our studies with N. glutinosa did not allow separation
of labdenediol and sclareol synthase activities (Guo & Wagner,
1995b), but recent results (unpublished) suggest that these activities
are due to separate enzymes. Thus, the tobacco diterpene synthases
appear to be members of a growing family of terpene cyclases
catalyzing formation of mono, sesqui, and diterpenes, the largest
group of plant secondary products.
Exudates are generally found in greater amounts on the lower surface
of leaves and are often higher on midribs, stems and sepals than on
leaves. The amounts of diterpenes produced by various tobaccos can
vary greatly. Lines such as TI 606 may produce as much as 151 and
13 µg/cm2 leaf of CBT-diols and -ols, respectively (Severson, et al.,
1985b). Line TI 1068, which we have used extensively to study the
enzymology and localization of exudate, yields strong biosynthetic
activity, yet produces only one-tenth as much CBT-ol and half as
much CBT-diol as TI 606. Other tobaccos, such as NFT, produce only
a trace of CBTs (Severson, et al., 1985a) and show low CBT-ol
biosynthetic activity (Guo & Wagner, 1995a). The labdenoids cis-
abienol and labdenediol ((12Z)-labda12,14-dien-8a-ol and (13E)-
labda-13-ene-8a,15-diol, respectively) are common exudate
components of Turkish type tobaccos. These diterpenes are generally
not present in burley, Maryland and dark type tobaccos, but are
common in cigar and primitive types. Severson reported significant
levels of these components in several nonUSA flue-cured types
(Severson, et al., 1985a). CBTs are thought to originate from N.
sylvestris, and cis-abienol and labdenediol from N. tomentiformis, the
presumed progenitors of Nicotiana species (Severson, et al., 1985a).
It is often useful to know approximately what percent of leaf dry wt
and crop yield is due to a given surface chemical. We have estimated
that in TI 1068 up to about 10% of leaf dry weight is exudate. From
this value one may predict that this tobacco could yield up to 225 kg
exudate per hectare (Wagner, 1991). The CBT-diols, SE, cis-abienol,
CBT-ols and labdenediols of TI 1068 account for about 62, 24, 12, 2
and 0.6% exudate weight, respectively. Generally speaking, CBT-diols
are the major components of tobacco exudates, even in lines where
exceptionally high levels of SE are found.
The approximate percent of CBT-diol and SE weights to exudate
weights were estimated for various tobacco types from the data of
Severson, et al. (1985a). Hydrocarbons and 1-docosanol were not
included as these are not exudate components. Also, tobacco types
having an unusual composition for their group were also excluded.
For USA flue-cured, burley, Maryland, cigar wrapper, dark fired, cigar
binder, and Turkish types, CBT-diols accounted for 95, 96, 96, 95, 96,
78 and 70%, respectively, of exudate weight. Some varieties may
contain substantial CBT-ols as well as diols (KY Black), while others
(like NFT) contain trace

 Page 296
levels of CBTs, very high cis-abienol, and high SE. The ratio of
carbon flow to cis-abienol versus CBTs may vary with developmental
stage and environment (Guo & Wagner, 1995a).
Various aspects of trichome exudate occurrence, surface deposition
and stability as affected by nitrogen supply, water stress, topping,
mechanical disturbance by rain or fog, light quality and intensity, soil
type, planting date, humidity and temperature have been assessed
(Severson, et al., 1985a). These aspects will not be discussed further
with the exception that new results regarding the role of humidity in
SE movement from the gland to the epidermal surface will be
considered below (Lin & Wagner, 1994a). As every tobacco producer
knows, water stress increases trichome exudate (gum) formation. This
has been quantified for several tobaccos (Severson, et al., 1985a).
There does not appear to be any evidence for 'induction' of exudation
as a result of insect or pathogen attack as occurs, for example, in
phytoalexins. Perhaps this is because exudates are already present (in
mature tobacco) as a first line of defense, while phytoalexins are
produced largely due to challenge (induced).
Sugar Esters (SE)
The second major group of tobacco surface chemicals is the SE. These
components are found in glandular trichome exudates of many plants
of the family Solonaceae, but apparently not outside this family. They
may occur as sucrose esters (i.e. in most N. tabacum, in N. glutinosa,
N. plumbaginifolia, N. langsdorfii and Petunia), as glucose esters (i.e.
N. miersii) or as mixtures (i.e. N. debneyi, N. cavicola, N. noctiflora,
etc.) (Cutler, et al., 1992). Potato species may contain sucrose or
glucose esters and Datura and tomato, glucose esters (Steffens &
Walters, 1991).
In Nicotiana, SE mixtures are well characterized and are grouped into
over eight types of acetylated or nonacetylated 2,3,4-tri-O-acyl-a-D-
glucopyranoside-a-D-fructofuranosides, where acyl groups are C3 to
C8 straight or branched (iso-or anteiso-branched) aliphatic acids
(Cutler, et al., 1992; Lin & Wagner, 1994b). While all known
Nicotiana sucrose esters contain three acyl groups on the glucose
moiety, the nature of groups occurring at specific positions (2, 3 or 4)
can apparently vary, as can the nature of mixtures of acyl groups
found in an SE type. Also, the number and position of acetyl groups
occurring in SE containing them may vary (C6 of glucose or C2 or C3
of fructose). Thus, the number of distinct SE structures produced by
members of the Nicotiana is tremendous. The biological significance
of this species-specific diversity is not known.
The use of selective NMR techniques to achieve regiochemical
assignment of acyl groups to specific positions in SE is beginning to
expand our understanding of the complexity of SE chemistry to the
next level (Matsuzaki, et al., 1992; Shinozaki, et al., 1992; King, et
al., 1993). Sugar esters and their acyl constituents are generally
characterized using GC or GCMS (Arrendale, et al., 1990; Severson,
et al., 1984) of trimethylsilyl derivatives or by HPLC of underivatized
molecules (Kandra & Wagner, 1988). Partial separation of SE group
members has been achieved using LH-20 gel filtration
chromatography and GC (Severson, et al., 1985c). Characterization of
acyl group short, medium and long chain fatty acids as free acids by
C18 HPLC (Kroumova & Wagner, 1995) has permitted discovery of
their unusual biosynthetic origin.
Biosynthesis of SE is shown to occur in trichome gland cells of
tobacco and tomato, but not in epidermal or subepidermal cells
(Kandra, et al., 1990). The sucrose or glucose moiety may be derived
from invertased sucrose transported to the gland and/or CO2 since
isolated glands are capable of synthesizing SE sugar (and acyl)
moieties from bicarbonate (Kandra & Wagner, 1988). Acyl moieties
may be straight or branched (iso or anteiso) and in tobacco and
petunia are derived by the alpha ketoacid elongation pathway (a-
KAE), a modified form of branchedchain amino acid metabolism
(Kandra, et al., 1990; Kroumova, et al., 1994). They are not formed
via fatty acid-synthase or -elongase mediated metabolism. Only in one
published case is an unsaturated acyl constituent shown, the detection
of a methyl C4 glucose ester acyl group having one double bond in N.
miersii (Matsuzaki, et al., 1989). This finding has been confirmed by
different methods (Kroumova & Wagner, unpublished data). It is
speculated that this group is formed by conversion of a-KAE-derived
2-methylbutyryl CoA via 2-methyl-branched chain CoA
dehydrogenase (unpublished results). Esterification of acyl moieties of
glucose esters in tomato is reported to occur via UDP glucose fatty
acid transglucosylation and transacylation (Ghangas & Steffens,
1995).
The basis of the diversity in acyl group chain types (iso-versus
anteiso-branched versus straight chain) in SE mixtures (particularly in
tobaccos) is not known. It has been speculated that it may be due to
substrate specificity of alkyl isopropyl malate synthase, a key enzyme
of a-KAE (Wagner, 1992). The basis of chain-

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length diversity is undoubtedly due to some chain termination
mechanism in a-KAE, but this is not characterized. Enzymes (and
their genes) for SE formation have not yet been purified and
characterized. The basis of the specificity of acyl substitution on the
glucose moiety (position 2, 3 or 4) or acetyl substitution of C6 or of
acetyl substitution at C2, 3 of fructose in certain sucrose esters is not
known. Plant breeding studies suggest that esterification is controlled
by one gene while acyl group formation is regulated by others
(Severson, et al., 1985a).
The amounts of SE relative to other exudate components in various
tobaccos have already been noted. They are most abundant in Turkish
types, but high levels can be found in certain flue-cured, primitive and
pale yellow types. They are least common in burley and Maryland
types (Severson, et al., 1985a). Turkish types are enriched in the 3-
methylvaleryl acyl group that is released on pyrolysis to give 3-
methylvaleric acid, the principal 'Turkish note' of their smoke
(Severson, et al., 1985a).
A recently developed procedure was used to stain SE on tissue
surfaces with rhodamine B to monitor SE amount, location (around
glands versus on the surface) and interaction with insects walking on
the surface (Lin & Wagner, 1994a). The method is simple,
quantitative but not selective for SE type. This method was compared
with one based on physical separation of gland-associated versus
below-the-gland associated SE and similar results were obtained (Lin
& Wagner, 1994a). Observations indicate that SE escape containment
around the gland (perhaps through pores in the cuticle) and flow down
the trichome stalk into depressions around epidermal cell cuticles
(Fig. 8.31). Migration of sucrose esters is greater than that of CBTs
and is enhanced by high humidity. Thus, migration is correlated with
compound polarity (Lin & Wagner, 1994a). Migration of exudate may
be particularly important to their presumed function for the plant as
anti-fungal and anti-bacterial agents. Aphids allowed to walk on
rhodamine-stained surfaces accumulate SE, first on their legs then
over their entire bodies (Lin & Wagner, 1994a and unpublished data).
Quantitation of rhodamine-stained SE by fluorimetry is so sensitive as
to allow detection of SE on a single aphid (unpublished results).
Sucrose esters, like CBTs appear to be stable on the undisturbed green
plant until senescence (Lin & Wagner, 1994a). However, turnover of
CBTs has been reported in tobacco shoot cultures (Marysia, et al.,
1993), and degradation (oxidation) is reported to occur in the field
(Severson, et al., 1985a). Unlike CBTs, they

Fig. 8.31
Migration of sucrose esters from trichome-gland
containment to the leaf surface as observed by
histochemical staining. Tl 1068 leaf was exposed
to high relative humidity and stained with
rhodamine B (35) to reveal SE migration from the
gland, down the trichome stalk then on to the
leaf in depressions between epidermal cells.
T = gland above the plane of focus, Ro-SE =
rhodamine B-SE in depressions between cells,
GC = guard cell.
are apparently also stable, post-harvest. When pyrolyzed, acyl groups
are released as volatile acids, and these contribute to flavor and aroma
(see following section). Levels on the green leaf may be decreased by
rain washing (Severson, et al., 1985a).
Surface Waxes
Cutin and surface waxes (the main components of the cuticle) and
their associated components comprise the third major group of
tobacco surface chemicals. These occur in all terrestrial plants, but
plants vary greatly in cuticle composition. The cuticle protects the
plant from desiccation, serves as a barrier against pests and
herbivores, and may function in light reflection, frost hardiness, etc.
(von Wettstein-Knowles, 1993). This covering is thought to be
continuous on the plant aerial surface (including surrounding the
trichome stalk and gland). It has been described as having three
layers: an outer wax layer lacking cutin, a thinner, amorphous primary
layer containing cutin and embedding wax and an inner, thick
reticulate secondary cuticle, also containing cutin and embedding
waxes (Bukovac, et al.,

 Page 298
1990). Elucidation of the cuticle physical structure has been hampered
by the insolubility of some components in both aqueous and organic
solvents. However, 13C-NMR is beginning to reveal structural
features (Pacchiano, et al., 1993).
Cutin, a principal component of the cuticle, is a polyester composed
mainly of cross-esterified hydroxy and epoxy fatty acids embedded in
wax. The outer layer (epicuticular) is generally found to be a mixture
of odd and even numbered, long chain paraffinic hydrocarbons, wax
esters (these two are most abundant), fatty alcohols, ketones and free
fatty acids (von Wettstein-Knowles, 1993). In some plants, small
amounts of flavanones, triterpenes, phenolics, polysaccharides and
polypeptides are thought to be associated with the cuticle (Bukovac, et
al., 1990). The outer wax layer may be amorphous, semicrystalline or
crystalline. This layer imparts texture and color to the plant surface.
Distinctly structured projections of surface wax can also contribute to
interactions with light in some plants (Pacchiano, et al., 1993). The
cuticular layer has been described as a high capacity ion exchange
polymer with high affinity for Ca++ over K+ or Na+ and an isoelectric
point of about 3 (Bukovac, et al., 1990).
Tobaccos of all commercial classes (i.e., flue-cured, burley, etc.) are
shown to contain about constant levels of methylene-chloride-
extractable surface aliphatic hydrocarbons (Severson, et al., 1985a).
These are C25 to C36, normal, anteiso-or iso-branched and are
generally present in a quantity up to 25% that of extract weight. This
extract would not contain cutin and some components of its
embedding wax. A constant, but low level of the fatty alcohol, 1-
docosanol, is common in tobaccos. Low levels of many chemically
distinct, high molecular weight wax esters are also common
(Arrendale, et al., 1988).
Arrendale, et al., (1988) studied the wax ester fraction of NC 2326
leaf in detail, as wax ester components have been reported in cured
leaf and in smoke. Surface waxes were washed from leaf surfaces
using methylene chloride and partitioned between 80% methanol in
water and hexane. Exudate components largely partitioned into the
methanol phase (Severson, et al., 1985c). The hexane phase contained
wax esters, hydrocarbons and fatty alcohols. These three were
separated by chromatography on silicic acid. The wax ester fraction
obtained was further purifed using Sephadex LH-20. Esters were
saponified and the products analyzed by GC as trimethylsilyl ethers
(alcohols) or methyl esters (acids). Free fatty alcohols and alcohol
moleties of wax esters of C18 to C30 were found. Of these, normal
C22 and C20 components were the most abundant. The free fatty
alcohol, 1-docosanol, was the most abundant of this chemical class.
Acid moieties of wax esters were Cl2 to C30 with normal C14 being
the most abundant. About 25% of wax ester fatty acids were iso-or
anteiso. Branched acids in surface wax esters are also common in
grasses. A total of 172 distinct wax ester species ranging from C30 to
C52 and having normal-normal, normal-iso or normal-anteiso alcohol-
acid composition were identified in NC 2326. This diversity may be
due to a lack of specificity in the transacylase reaction forming esters
(see below). This study in NC 2326 is a rare example of an in-depth
examination of surface waxes of tobacco, or any plant.
Hydrocarbons of the plant cuticle are formed by elongation of
preformed acids, then loss of the carboxyl carbon (decarbonylation)
(Kolattukudy, 1987). Wax esters are derived by esterification of
primary alcohols with fatty acids putatively derived from fatty acyl
CoAs or phospholipids. Exogenously supplied short-and long-chain
acids are efficiently incorporated into hydrocarbons and wax esters as
are valine and iso-leucine (Kolattukudy, 1987). Thus, it is thought that
branched acids of wax esters are derived by fatty-acid-elongase-
mediated elongation of short branched primers. However, detailed
analysis of the origin of carbons in products of precursor feeding
experiments leading to these conclusions was not made. And, with the
discovery of the role of a-KAE in SE acyl group synthesis, the
possibility exists that particularly branched-chain surface lipid
components may be derived in part or wholely by a-KAE.
While elongases and associated enzyme complexes involved in
synthesis of cuticle components are found to be associated with
particulate fractions of epidermal homogenates, the subcellular site of
cuticle component biosynthesis is not confirmed (von Wettstein-
Knowles, 1993). The endoplasmic reticulum is often suggested to be
involved, but enzymes are often enriched in a 3000 × g fraction,
suggesting that they may be primarily in the cell wall. This is also true
of enzymes that synthesize suberin, a polyester related to cutin that is
found in various anatomical locations in plants (i.e. casparian strip in
roots) (von Wettstein-Knowles, 1993). It is particularly tempting to
speculate that in wax ester formation, elongated alcohol and acid
moieties are formed inside the cell and that transacylation occurs in
the cell wall. The great diversity in wax ester types found in tobacco
suggests random transacylation, perhaps like random polymerization,
as in the case of wall-peroxidase-catalyzed lignin formation.
Mutants in rice, Brassica, Zea, Arabidopsis and Sorghum, having
altered cuticle structure and composition

 Page 299
are providing insights into the biosynthesis, structure and function of
the plant surface waxes (Hoffmann-Benning & Kende, 1994; Jenks, et
al., 1994).
Volatile Compounds
The fourth class of surface chemicals to be considered is the volatiles,
known to be produced in both leaf and floral tissues. These
compounds are undoubtedly produced by epidermal layer cells and
should be considered surface constituents. In tobaccos many (up to
40) volatile terpenes, aromatic and aliphatic compounds were
recovered from flowers by head space analysis (Loughrin, et al.,
1990). High boiling volatile compounds were trapped on the porous
polymer Tenax (2,6-dipheyl-p-phenylene oxide), desorbed, then
analyzed by capillary GC. A lesser number (total about 12)
compounds from all three chemical classes were found in leaf head
space (Andersen, et al., 1988). Burley (KY 14), pale yellow (TI 1068),
the primitive type (TI 1112) and the mutant line (TI 1406); all
volatilized the sesquiterpene caryophyllene as a major floral volatile.
Of these tobaccos, only TI 1068 produced the monoterpene alcohol
linalool as a major component. Caryophyllene was the second most
abundant component. Linalool is a common floral volatile in plants
(Kelsey, et al., 1984; Pichersky, et al., 1994). Since TI lines 1112 and
1406 are glandless and glandular-but-not-secreting, respectively, it
was concluded that exudate-secreting trichome glands are not
responsible for production of volatiles. The question of whether
hydathodes may be involved is, however, not yet answered. While
caryophyllene was found to be a minor and neophytadiene a major
component in leaf headspace in tabacums, the reverse order of
abundance was found with flowers (Loughrin, et al., 1990). Yield of
volatiles (rates of production) in tobacco flowers was estimated to be
in the mid-range of that reported for other plant species and was 30 to
100 times greater than amounts observed with tobacco leaves
(Loughrin, et al., 1990).
It is difficult to compare carbon flow through volatile versus
trichome-exudated components, but using certain assumptions, it is
possible to calculate that, in tobaccos, floral volatile compounds may
be produced at 0.01 to 0.001 the rate of exudates. Floral volatile
production in N. suaveolens has been calculated to be about 1 µg/g
fresh weight per 20 hours (Loughrin, et al., 1990) or 0.05 µg/g fresh
weight per hour. While few studies have carefully examined rates of
trichome exudate formation, one may predict production of 80 µg
exudate/cm2 per week or 1 µg/cm2 per hour (assuming synthesis
during the day only, so per 80 hours). Such rates were observed for
mature leaf of several tobaccos in a study of bud leaf CBT synthesis
versus time after transplanting (Severson, et al., 1985a). This is the
equivalent of 50 µg/g fresh weight per hour or 1000-fold the rate of
measured floral volatile production. However, volatiles are functional
at low levels and are extremely important for pollination (Dobson,
1993).
Apparently the sites of biosynthesis of volatiles in tobacco leaves and
flowers are not known. In flowers of other plants, various tissues
(petal, stamen, stigma, etc.) may participate in volatile production, or
this may primarily be located in specific tissues. Also, specialized
osmophores (extensively studied in the Orchidaceae) may be
localized in certain tissues and sites within these tissues for the
apparent purpose of massive volatile production to attract and perhaps
guide pollinators (Pridgeon & Stern, 1983). It has been suggested that
specific osmophores may produce specific components. Alternatively,
sites of volatile production may be more generally dispersed and may
deliver their products through the cuticle for volatilization (Pridgeon
& Stern, 1983) or to accumulate as droplets on the surface.
As is common in most plants, certain tobacco volatile emissions (like
benzyl alcohol) follow a circadian rhythm (Loughrin, et al., 1991).
Others, such as caryophyllene, do not. In some tobaccos there is
temporal synchrony of emissions with pollinator activity (Loughrin, et
al., 1991). Glycosidically bound volatiles of N. sylvestris and N.
suaveolens (principally aromatic compounds) found in homogenates
of flowers may be precursors to volatilized aglycones (Loughrin, et
al., 1992). As can be seen from the cited references, detailed study of
volatiles from vegetative tobacco was just recently begun.
Functions of Surface Chemicals to the Growing Plant
The role of the cuticle in plant-water relations is central to the growth
and development of all terrestrial plants (von Wettstein-Knowles,
1993). The cuticle may also deter herbivory or stimulate oviposition
(Espelie, et al., 1991). Perhaps the next most important role that
surface chemicals serve to the plants is a defensive one against insect
and microbial damage. Trichome exudate constituents are key
components in this role in tobacco and other Solanaceous plants.
Several reviews have

 Page 300
discussed in detail insect-and microbe-interactive properties of
tobacco trichome exudates and surface waxes. The reader is referred
to these for details (Cutler, et al., 1986; Espelie, et al., 1991;
Severson, et al., 1991; Cutler, et al., 1992; Severson, et al., 1993) and
to the section herein by Jackson, Semter and Massee. Here only a
brief update summary is offered. Also, some little discussed aspects of
surface chemical function are covered.
Exudates may physically immobilize or slow insect activity on the
surface by virtue of their stickiness. They may be insecticidal or anti-
microbial or, conversely, they may participate positively in an insect's
life cycle. A much studied example of the last is the stimulation of
tobacco budworm (Heliothis virescens) moth oviposition in tobacco.
In this case, CBT-ols and -diols, and to a lesser extent certain SE
types, appear to be active agents, while labdenoids, hydrocarbons, 1-
docosanol and wax esters, and certain other SE show lower or little
activity (Severson, et al., 1993). These same ovipositional stimulants
are, however, toxic to larval growth. Low levels of CBTs and SE are
correlated with susceptibility to flea beetle (Epitrix hirtipennis) and
Japanese beetle (Popillia iaponica) damage (Severson, et al., 1985a).
Possible direct effects of surface chemicals on susceptibility to virus
damage are little studied.
From recent studies, CBT-ols and certain SE types are implicated in
aphid resistance (Severson, et al., 1993). SE of N. gossii, N.
benthamiana and N. umbratica are shown to afford resistance to the
soft-bodied insects (the tobacco aphid, Myrzus nicotiana, the
greenhouse whitefly, Trialeuroides vaporariorum, Westwood and the
sweet potato whitefly, Bemisia tabaci, Genadius) that are important
pests of many crop plants (Buta, et al., 1993; Severson, et al., 1993).
In one study, CBT-ols, cis-abienol or nicotine were 20 times less toxic
to aphids than N. gosseii SE (Severson, et al., 1993). The unique
feature of SE from N. gosseii and certain other tobaccos is that their
acyl groups are composed, in part, of branched, medium chain
(methyl-hexanoyl, -heptanoyl, -pentanoyl) constituents (Shinozake, et
al., 1991; Matsuzaki, et al., 1992; Severson, et al., 1993). Acyl groups
of cultivated tobaccos are predominantly branched, but are short chain
(C3 to C5). It had been established earlier by Tingey and colleagues
that the insect-retardant activity of Lycopersicon pennellii (primitive
tomato) is due to the occurrence of 8-methylnanoyl groups in
trichome-exudated glucose esters (Steffens & Walters, 1991).
Similarly, long-chain acyl groups of SE from Solanum berthaultii
(primitive potato) are implicated in insect resistance in this plant
(King, et al., 1988). It is well known that medium chain length and
some types of branching and conjugation (esterification) in fatty acids
are among the properties that often correlate with optimal anti-
microbial properties of fatty acids. Cultivated species of tobacco and
tomato selected for yield and other characteristics may have lost the
capacity to elongate SE acyl groups in a-KAE and thereby lost natural
pest and disease resistance. Patents have already been issued for the
concept of using resistance-conferring tobacco SE as
environmentally-safe, natural-product pesticides for use on other
crops.
The effects of tobacco exudate components on the fungus
Peronospora tabcina (blue mold) have been assessed (Menetrez, et
al., 1990; Kennedy, et al., 1992). While data are somewhat
contradictory, CBT-diols and cis-abienol appear to show anti-fungal
activity. Sclareol, found in certain N. glutinosa varieties, is reported to
be an inhibitor of rust fungi and its 2-keto-8(18)-ene derivative is
reported to be an inhibitor of powdery mildew (Banthorpe, et al.,
1990). Growth of various gram-positive and gram-negative bacteria in
the presence of exudate components has been assessed by several
groups (Cutler, et al., 1986; Cutler, et al., 1992; Kennedy, et al., 1992;
Chortyk, et al., 1993; Kennedy, et al., 1995). Generally, SE having
longer, branched acyl groups are most effective against gram-positive
species (Chortyk, et al., 1993). They are generally inactive against
gram-negative types, but this distinction is not yet clear (Cutler, et al.,
1992). Synthetic SE with long chain acyl groups in combination with
sorbate and propionate afford increased anti-mitotic activity and cause
lower aflatoxin formation in Aspergillus parasiticum (Marshall &
Bullerman, 1986). Thus, SE undoubtedly provide for pest and
microbe resistance functions on the tobacco surface, and they may
have potential for commercialization as pesticides. Synthetic SE are
already marketed for this purpose (Smith & Stow, 1984). Tobacco
produces complex SE that are difficult to synthesize chemically. Some
of these may have unique pest-resistance properties. The potential of
high-biomass, highly-exudated, modified tobaccos as 'factory plants'
for producing commercially Valuable exudate chemicals has been
discussed (Wagner, 1991). It is also noteworthy that so-called
'designer fats' are synthetic SE.
It has been reported that tobacco SE, CBTs and labdenoids have
growth-regulating properties (Cutler, et al., 1986; Shinozaki, et al.,
1991; Cutler, et al., 1992). This is concluded from in vitro
experiments using the etiolated wheat coleoptile elongation system
and from assessments of seed germination inhibition and shoot

 Page 301
bleaching of barnyard grass and germination inhibition of hemp
sesbania. Growth inhibition of wheat coleoptiles was correlated with
SE 3-methylvaleryl content. Pure 3-methylvaleryl glucose SE was not
effective, suggesting the importance of a sucrose ester or esterification
at specific sites on the sucrose moiety (Cutler, et al., 1986). While
these observations are interesting and should be extended to studies of
effects on young leaves, it is premature to imply that SE may function
as growth regulators on the growing plant. The term 'effector' rather
than 'regulator' may be more prudent at this time. SE may have
usefulness as growth regulators in a commercial sense.
Little is known about the fate or modes of toxicity of SE in insects or
microbes. As noted earlier, aphids walking on the tobacco leaf surface
can rapidly become contaminated with exudate. We have monitored
the fate of radiolabeled SE supplied to fifth instar tobacco hornworms
in food and recovered intact SE from hemolymph and carcass
(Lomba, Dahlman and Wagner, unpublished data). Thus, these
molecules appear to pass this insect's gut intact. It is not known if
their detergent properties are the cause of observed growth inhibition.
Summary
The surface chemistry of green tobacco is largely elucidated. The next
level of understanding in this area is perhaps to determine the
regiochemistry of acyl substitution in specific SE. Better methods will
be needed to separate complex mixtures of SE molecules.
Understanding of the biochemistry leading to surface chemicals, on
the other hand, is in its infancy. Enzymes and pathways for
cembrenoid and labdenoid diterpenes are now being elucidated but no
genes for these are yet available. The pathway for SE acyl group
formation has been discovered, but enzymes involved are not yet
studied. We know little about the subcellular sites of surface chemical
formation. Improved knowledge in this area is needed to support
future efforts to manipulate surface chemistry by genetic engineering.
A challenge for the immediate future will be to isolate enzymes and
genes and use these to manipulate surface chemistry to improve
natural pest resistance, perhaps improve flavor and aroma qualities
(Wagner, 1991; Wagner, 1992; Gadani, et al., 1995) and to exploit the
commercial potential of tobacco as a 'factory plant' for producing
easily recovered commercial products. New variety development
continues and can benefit from availability of new tools to monitor
traits (antibodies, genes).
It is unfortunate for tobacco science in the USA that the
'biotechnology era' has coincided with the loss of USDA tobacco
laboratories, a bias against using federal funds to support tobacco
science (even to some extent basic science) and the movement of
several excellent tobacco scientists to other crop systems. At the same
time, more work is needed to define the roles of surface chemicals on
insects and microbes and to elucidate the biochemistry and molecular
biology of surface chemical production. Also, much more study is
needed to explore possibilities for modifying surface chemicals, post-
extraction (Arnarp, et al., 1993) if the commercial potential of surface
chemicals is to be exploited.
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8D
Relationship between Leaf Chemistry and Organoleptic Properties of
Tobacco Smoke
W.W. Weeks
North Carolina State University
Raleigh, North Carolina, USA
Introduction
The use of tobacco in North America precedes the arrival of
Christopher Columbus because native Americans were already
obtaining pleasure from smoking and chewing native tobacco
(Massie, 1981). Tobacco is now grown around the world and the
tobacco industry is among the world's leading enterprises. Consumer
products manufactured from tobacco include cigarettes, cigars, pipe
tobaccos, snuff and chewing tobacco (Leffingwell, et al., 1972).
Tobacco quality is influenced by weather, soils, management and
curing (Dawson, 1952). Discussion of the organoleptic properties will
be directed primarily to the relationship of certain chemistries from
tobacco on sensory perception of smoke.
In the USA, tobacco is classified according to its curing regime. Each
class is further divided into types, and type is divided into grades.
Grade defines stalk position, color, uniformity, quality, body and other
physical attributes of the leaf (Barnard, 1959; Tso, 1990). The three
major classes grown in the USA include flue-cured, burley and
Maryland tobaccos. Oriental or Turkish is an important class of
tobacco grown in the Balkans, notably Greece, Turkey and Former
Yugoslavia. Flue-cured comprises the largest percentage of tobacco
grown and is used primarily in the manufacture of cigarettes. Smoke
from flue-cured tobacco is sweet and aromatic with a slight acidic
taste as a result of high levels of reducing sugars in the cured leaf
(Leffingwell, et al., 1972). Burley, the second largest percentage of
tobacco grown in the USA, is an aircured tobacco that has excellent
smoking characteristics. It is less dense and bulkier than flue-cured,
darkbrown to red in color and produces a more basic smoke and a
taste described as winey, chocolate, nutty and ammonia-like
(Leffingwell, et al., 1972). Maryland is an aircured tobacco similar to
burley but milder in smoking characteristics. Oriental tobacco is
aircured and characterized by small leaves with aromatic properties
that contribute a pronounced taste and aroma to blended cigarettes
(Leffingwell, et al., 1972).
American cigarettes consist of tobacco blends (Green, 1977). Blends
are composed of a mixture of flue-cured, Oriental, burley and/or
Maryland tobaccos. Flue-cured generally comprises the largest part of
a blend; however, the amount of each tobacco used in the blend
depends upon the smoke characteristics the manufacturer is trying to
achieve. Casings and flavors are used in manufacturing cigarettes to
maintain consistency of the blend and improve smoking
characteristics (Leffingwell, et al., 1972; Bertin, 1979). The
manufacturer is interested in maintaining consistency and improving
quality of blends over time as economically as possible. To do this, it
is necessary to establish a correlation between the properties of
tobacco and tobacco smoke (Green, 1977).
Each compound produced in smoke is a potential contributor to the
organoleptic properties. However, the concentration of many
compounds that have been identified is too low for even the most
sensitive smokers to detect and identify. The chemical composition of
a tobacco leaf varies with the class of the leaf and the stalk position.
Many compounds can be detected analytically with very sensitive
techniques at very low concentration, but smokers cannot readily
identify the taste of these compounds. At the same time, there are
some compounds in smoke that are difficult to quantify but the
smoker is able to detect them readily on the palate and tongue at very
low levels (Bertin, 1979). Compounds are transformed from tobacco
to smoke during pyrolysis by:
(1) direct transfer from volatilization, although only a third or less of
the concentration of a compound found in tobacco transfers directly
from tobacco to smoke (Wakeham, 1972),
(2) transfer from the tobacco to the smoke by mass transfer, and
(3) generation of new compounds from pyrolysis.

 Page 305
During smoking, the smoker is subjected to favorable sensations of
flavor, aromas and feelings generated from irritation in the aerosol
from combustion of tobacco. Taste of smoke results from constituents
in the aerosol that dissolve in the saliva, making contact with the taste
buds, stimulating the nerves and sending an impulse to the brain
prompting the memory to recognize the taste. There are four primary
tastes which are recognized: sour, sweet, salty and bitter. The many
compounds in the aerosol interact synergistically, diluting the effect of
individual compounds and giving a sensation unlike the four basic
tastes. Nevertheless, detectable concentration of a substance in the
aerosol that the smoker can identify in the presence of other
compounds is described as the threshold of that compound
(Abadallah, 1970). Pleasing tastes give smoking satisfaction, and
unpleasant tastes result in irritation as well as dissatisfaction in
smoking (Bertin, 1979).
Olfaction and taste are very closely related. It is the aroma of smoke
that tantalizes and encourages smoking. Smoke that is free of odor is
flat and tasteless. Odor is perceived by millions of tiny hairs which
cover the epithelium located in the roof of the nasal cavity. Individual
aromas are detected by an unsolved mystery of science which cannot
be explained but plays an important role in smoke assessment. The
characteristic odors perceived by the olfaction system are floral,
putrid, pungent, acrid and camphorous (Meilgaard, et al., 1987). The
palate and the olfactory senses are in close proximity and the
combined function of these two systems gives a true impression of
smoke flavor.
Leaf and Smoke Components
The data in Table 8.18 (Weeks, 1985) show the categories of a number
of compounds and indicate their ranges in the cured leaf. By
reviewing this list, it is somewhat easier to understand the number of
individual compounds produced in the smoke from different tobaccos.
Flue-cured tobacco is higher in carbohydrates and lower in
nitrogenous compounds than air-cured leaves. Burley is higher in
mineral constituents than flue-cured and Oriental types. The latter
explains why air-cured tobaccos produce the highest ash content
(Demole & Dietrich, 1977). The chemistry of Oriental is intermediate
to that of air-cured and flue-cured in composition. For instance,
Oriental is lower in nitrogenous constituents than burley and
Maryland types and lower in carbohydrates than flue-cured, but it is
Table 8.18 Tobacco leaf
components.
(percentages)
Waxes and wax esters 00.6601.20
Solanesol and esters 00.8002.00
Organic acids 03.0007.67
Polyphenols 00.7505.70
Reducing sugars 00.8025.00
Non reducing sugars 01.0005.00
Starch and pectins 00.0008.00
Nicotine 00.2804.00
Amino acids 00.2503.00
Cellulose and lignin 25.0028.50
Volatile oils 00.2501.00
Protein 01.0003.00
Water (free and bound) 11.0024.00
Source: Weeks (1985).

considerably higher in resinous terpenes and volatile acids than burley

and flue-cured (Leffingwell, et al., 1972).
Leaf components, upon combustion, produce a multitude of
compounds that influence the taste and perception of smoke (Table
8.19) (Roberts, 1988). Roberts (1988) alluded to compounds found in
the smoke according to functional group. Although over
Table 8.19 Compounds found in smoke.
Functional No in No in No common
group Tobacco Smoke to both
Carboxylic 450 69 140
acids
Amino acids 95 18 16
Lactones 129 135 39
Esters 529 456 314
Amides and 205 227 32
imides
Anhydrides 10 10 4
Aldehydes 111 106 48
Carbohydrates 138 30 12
Nitriles 4 101 4
Ketones 348 461 122
Alcohols 334 157 69
Phenols 58 188 40
Amines 65 150 37
N-Hetrocycles:
Pyridines 63 324 46
Pyrroles and 9 88 3
indoles
Pyrazines 21 55 18
Nonaromatic 13 43 7
Ethers 53 88 15
Hydrocarbons 184 429 114
Inorganics and 105 111 69
metals
Source: Roberts (1988).

 Page 306
4000 compounds have been identified in tobacco smoke, relatively
few contribute to smoke taste.
a
Carbohydrates
Carbohydrates comprise 40% or more by weight of the lamina of
tobacco leaves (Table 8.18). From the carbohydrate fraction of the
leaf, glucose, fructose, arabinose and xylose have been found in small
quantities in cigarette smoke (Stedman, 1968). Studies are also
available on analysis of cellulose, pectins and starch.
Sugars are important in the food industry; therefore, degradation
products from thermal treatments of sugars from food products have
been thoroughly investigated. Gas chromatography of chloroform and
methylene chloride extracts prepared from thermal degradation of
glucose and fructose at 175 to 275°C resulted in the production of
factones, ketones, cyclopentane derivatives and short chain carboxylic
acids (Ferretti, et al., 1970). Taste and perception of these compounds
have a similar influence on the smoker somewhat comparable to
flavor compounds produced by nonenzymatic browning reactions
(Green, 1977). Glucose and fructose heated to 840°C resulted in
production of aromatic hydrocarbons, furfurals and phenols. Acetone
and acetaldehyde were also produced from pyrolysis of glucose at
800°C (Ivanov & Ognyanov, 1966; Green, 1977). Either of these
compounds can be detected in small quantities and very readily
become irritants in smoke. Smoke studies with 14C glucose and
fructose have also shown that most of the compounds derived from
pyrolysis of glucose and fructose are recovered in the total particulate
matter (TPM) of cigarette smoke (Jenkins, et al., 1975; Green, 1977).
Furfurals and phenols have been shown to influence smoke taste and
aroma (Green, 1977).
Matsukura and Ishiguro (1986) isolated furan and furanone
compounds from flue-cured tobacco roasted at 150°C and applied
these compounds to cigarettes. They concluded that improved taste
and aroma were obtained from cigarettes made from compounds
isolated from flue-cured tobacco roasted at 150°C. Production of
chemical components from pyrolysis of simple sugars is well
documented; however, the most important effect is the one that
pyrolysis has on the pH of the smoke (Leffingwell, 1976). In the case
of flue-cured and Oriental tobacco, sugar is generally high, resulting
in thermal decomposition at temperatures comparable to the
combustion temperatures of amino acids producing ammonia from
air-cured tobacco, and the resulting pH of flue-cured and Oriental
smoke is, therefore, acidic. This pH effect results in reduction of
smoke strength and impact by modifying the effects of nicotine and
other bases in the smoke (Fenner, 1988).
It is difficult to conceive and identify the taste and perception of a
specific compound from among the many compounds found in
smoke. Taste and perception of a compound are established by
directly adding the compound to tobacco before smoking in a quantity
that exceeds the quantity of the compound found in leaf. This does not
give an accurate perception of the natural taste of the tobacco because
the concentration added far exceeds the amount found naturally in the
leaf (Kallianos, 1976). On the other hand, individual compounds have
been pyrolyzed in quantities greater than the amount found in tobacco
at different temperatures to determine the fate of individual tobacco
compounds. Experimental studies of additives have been enlightening
in that pyrolysis products derived from them were identified and this
has led investigators to identify precursors in tobacco to compounds
found in smoke. However, this process has been of little value when
associating the precursors of compounds found in smoke with quality
and perception of smoke (Chortyk & Scholtzhauer, 1973). Smoking is
the predominant means of consuming tobacco. Constituents in the
smoke are contributors to its organoleptic properties. Thirty percent or
more of leaf weight is composed of high molecular weight polymers
that produce largely water, CO2 and water-soluble compounds that
accumulate in the particulate matter of smoke. At the same time, some
of these products from the high molecular weight polymers influence
the organoleptic properties of the smoke (Obi, et al., 1970; Green,
1977).
b
Cellulose
Cellulose is the major structural constituent in the tobacco leaf. This
high molecular weight polymer is primarily responsible for integrity
of the leaf. During curing, aging and processing the cellulose content
of the leaf remains constant. Products from combustion of cellulose
result in roughly 12% of the weight of the pyrolysis products in the
main stream of smoke (Chortyk & Scholtzhauer, 1973). Products
begin to form from cellulose at 350°C and reach a maximum between
700 and 750°C. For the most part, phenols are the predominant
compounds produced from pyrolysis of cellulose. These compounds
are similar in taste to food flavors with smoky aroma and taste.
Combustion of cellulose produces b-D-glucopyranose and other
similar compounds characterized by caramel scorched aroma and taste
from smoking (Heath & Reineccius, 1986).

 Page 307
c
Lignin
Lignin, a high molecular weight polymer, contains 100 or more
aromatic units high in methoxyl groups. Lignin is the source of such
flavor compounds as vanillin, benzyl alcohol, 1-phenylethanol, 2-
phenylethanol and other aromatic compounds produced and distilled
into mainstream smoke (Green, 1977). Benzyl alcohol and l-and 2-
phenylethanol have been described as floral and fruity, contributing to
the smoothness of smoke. These compounds are important at low
concentrations but add bitterness at high concentrations (Roberts,
1988).
d
Pectins
Highly complex, water-insoluble carbohydrates are known as
protopectins which, upon partial hydrolysis, yield pectins or pectic
acids containing carboxyl and methoxy-carbonyl groups. Purification
of pectic acid and hydrolysis yields about 85% D-galacturonic acid,
arabinose, galactose, glucose, xylose and L-rhamnose. Pectic acid
produces a carboxyl group per each group of six carbons. Analysis of
data obtained from leaf analysis shows that the mid-rib is higher than
the lamina in free pectic acid and protopectin. Varietal differences
reported in pectins appear to be due to difference in analytical
techniques rather than actual differences that occur among tobaccos.
However, flue-cured has more pectin material than burley. Differences
in pectin material due to physiological differences in leaves and the
effect upon leaf quality have been studied. Shmuk (1953) indicated
tobacco quality was inversely proportional to pectin concentration.
Pyrolysis studies with isolated pectins are similar to pyrolysis of
starch (Green, 1977). However, pyrolysis of sugars and galactouronic
acid yields products similar to other carbohydrates. Fermentation of
pectins produces as much as 1.0% to 1.5% acetic acid, which affects
the taste of smoke by producing acrid taste and irritation.
e
Protein and Amino Acids
During curing, storage and aging many changes occur to proteins and
amino acids in the tobacco leaf. Without some available protein,
smoking quality of tobacco is very low. Some protein is needed in
tobacco to increase smoke strength and bitterness to improve taste of
smoke, at least for some smokers (Shmuk, 1953). Forty-four amino
acids have been identified in the tobacco leaf. At least 15 of these
have been identified in the smoke (Leffingwell, 1976). Most of the
information available on amino acids pertaining to combustion has
come from pyrolysis studies. Aliphatic amines are the predominant
compounds produced from pyrolysis of amino acids at 500°C
(Patterson, et al., 1969; Patterson, et al., 1971). Higher temperatures
result in production of 3-and 4-carbon compounds that recombine to
form aromatic compounds regardless of the amino acid (Patterson, et
al., 1969; Patterson, et al., 1971). Pyrolysis of proline, the most
abundant amino acid in flue-cured tobacco, at 800°C produces
pyrrole, several substituted pyrroles and an azalactone which have a
characteristic caramel, roasted-like aroma and a taste typically
associated with thermal browning reactions (Smith, et al., 1975).
Pyrolysis of b-hydroxy amino acids (serine) results in pyrazine
compounds that produce other important tobacco flavors (Green,
1977). Pyrazines are very important flavor compounds in very small
quantities that add nutty, coca and popcorn-like flavor to smoke
(Roberts, 1988).
The influence of sugars, carbohydrates and amino acids upon smoke
has been considered independently. Sugars naturally occurring in the
leaf during aging or sugars added during processing react with amino
acids to form Amadori compounds during storage and processing
(Leffingwell, 1976). Amadori compounds during pyrolysis produce
pyrazines and maltol, a flavor compound found in smoke that has a
characteristic sweet taste and is also a prominent enhancer of flavor.
The most prominent Amadori compound found in flue-cured tobacco
is 1-deoxy-l-L-prolino-D-fructopyranose. Formation of this Amadori
compound seemingly corresponds with the availability of substrate
and optimum conditions during storage. Other Amadori compounds
produced in flue-cured tobacco arise from the interaction of alanine,
threonine, phenylalanine and aspartic acid with glucose and fructose
(Leffingwell, 1976). According to Noguchi, et al. (1971), the amount
of Amadori compounds produced in the flue-cured leaf constitutes up
to 2% of the leaf's dry weight. Huyghues-Despointes, et al. (1994)
showed that the pyrolysis of 1-deoxy-l-prolino-D-fructopyranose at
350°C for 10 seconds produced 17 compounds. The profile obtained
by GC-MS appeared very similar to the profile obtained from
pyrolysis of a 50/50 mixture of proline and glucose under similar
conditions. Information is currently unavailable on the pyrolysis of
Amadori compounds at temperatures of 700 and 800°C.
Interaction of amino acids and sugars begins early in the post-harvest
storage of flue-cured tobacco. Amino acids undergo change
enzymatically and chemically by

 Page 308
decarboxylation and deamination to produce dicarbonyl compounds
that react with amino groups of amino acids with loss of CO2. The
degradation products undergo further change and interact to produce
pyrazine compounds which influence the taste and perception of
smoke (Patterson, et al., 1971). Pyrazine and substituted pyrazines are
formed by amino acid interaction when heated in the presence of
glucose and fructose in processed tobacco (Fujimaki, et al., 1973).
The transformation of sugars to lower molecular weight carbonyl
compounds during aging and storage occurs from oxidation. The
amine groups of amino acids, primary and secondary amines and
ammonia react with these carbonyl compounds to produce adducts
that, during pyrolysis, produce compounds associated with
nonenzymatic browning reactions that are important flavor
constituents in smoke (Yamaoto & Noguchi, 1973).
f
Tobacco Lipids from Cuticle and Surface of the Leaf
A large number of tobacco components have been classified under the
category of lipids and waxes. These compounds are found on the
surface of the leaf or as components of the internal structure of the
leaf. Cuticular waxes consist of a complex of hydrocarbons having
similar properties (Kolattukudy, 1968). These hydrocarbons consist of
straight and branched chains from 25 to 34 carbon atoms.
Hydrocarbons are synthesized in the epidermal layer as droplets from
16-and 18-carbon fatty acids and transferred to the surface in
crystalline form (Tso & Chu, 1970; Chortyk, et al., 1975). Waxes and
surface lipids function to protect the leaf surface and aid against the
loss of water from the intact leaf during growth. Chortyk, et al. (1975)
concluded that hydrocarbons and waxes transfer unchanged directly to
the smoke. Ivanov and Ognyanov (1966) reported that waxes have a
negative effect on the taste of smoke by adding harshness and mouth
coating. Although the presence of waxes in smoke has a negative
effect, these compounds help provide balance to prevent a bland
smoke (Davis, 1976).
g
Diterpenes
Diterpenes found in tobacco are of two classifications, duvanes and
labdanes. The acyclic duvanes are synthesized in the trichomes of the
leaf and accumulate on the surface as exudate during leaf maturation
(Chang, et al., 1985). Duvanes are predominant to labdanes in
tobacco. The predominant duvane found in tobacco is 4,8,13-
duvatriene-l,3-diol. This compound comprises up to 50% of the lipid
material found in an immature tobacco plant (Keene & Wagner,
1985). However, the concentration decreases as the plant matures.
Roberts and Rowland (1962), Chang, et al. (1985) and Weeks and
Seltmann (1986) have shown the relationship of this compound to
aroma of tobacco, primarily due to degradation products formed by
oxidation during senescence and curing. Oxidation products of
duvanes are solanone, oxysolanone and branched chain volatile acids
which contribute to smoke flavor associated with burley (Demole &
Dietrich, 1977). Solanone and oxysolanone contribute a ketonic taste
to smoke. Although duvanes are washed off and lost during growth,
breakdown products remaining after curing are important natural
flavorants.
Labdanes are di-and tricyclic diterpenes that are more common to
Oriental than to flue-cured or burley tobaccos. Oxidative breakdown
of abienol during curing produces a number of bicyclic compounds
that impart a pleasant cedary-like flavor and aroma to smoke (Enzell,
1976).
h
Internal Lipids
Trisesquiterpenes are found endogenously in tobacco, and diterpenes
and sesquiterpenes occur on the surface of the leaf. Neophytadiene,
the most predominant diterpene, is found in large amounts compared
to other volatile and semivolatile compounds in the volatile oils of
flue-cured and burley tobaccos. Neophytadiene is associated with
smoke quality, because it transfers directly into the smoke in
appreciable quantities (Chortyk, et al., 1975). A soothing and
smoothing effect upon smoke has been associated with neophytadiene
(Green, 1977). Greater quantities of neophytadiene are found in the
leaves from the upper part of the plant and it occurs in larger amounts
in flue-cured than in air-cured tobaccos (Davis, 1976). At higher
temperatures pyrolysis occurs with neophytadiene to produce smaller
components. However, a large part of neophytadiene in a sample
distills into the smoke (Davis, 1976; Green, 1977).
Solanesol, the major trisesquiterpene alcohol, is found as a free
compound, as an acetate and as sterol esters in cured tobacco leaves. It
is found in higher quantities in flue-cured than in air-cured tobaccos.
The concentration of solanesol increases with curing (Smith, 1980).
Although there is no direct association with solanesol and tobacco
quality or tobacco aroma, secondary benefits of solanesol are obtained
from

 Page 309
diterpenes produced from pyrolysis during smoking (Grossman, et al.,
1962; Davis, 1976).
Fatty acids are another category of lipids found in the tobacco leaf.
They have been studied in the immature green leaf, senescencing
leaves and cured leaves by stalk positions but most have been found in
cigarette smoke (Kolattukudy, 1968; Tso & Chu, 1970; Davis, 1976).
Two classes of fatty acids are lower molecular weight polar fatty acids
and high molecular weight nonpolar fatty acids, which vary
quantitatively with the type of tobacco studied. Generally flue-cured
produces higher quantities of high molecular weight fatty acids than
burley and other air-cured tobaccos. Most of the high molecular
weight saturated aliphatic fatty acids, 12 carbons or higher, add a
waxy, fatty and smoothing taste to smoke. Otherwise, high molecular
weight unsaturated fatty acids (linoleic and linolenic) add harshness to
smoke (Davis, 1976). The low molecular weight 5-and 6-carbon fatty
acids contribute to flavor and aroma of cigarette smoke and are
generally considered associated with Oriental tobacco and the 'dirty
sock' aroma perceived in blended cigarettes. Isobutyric, isovaleric,
valeric and b-methylvaleric acid are short chain aliphatic fatty acids
associated with sweet, winy, buttery, nutty and fruity tastes in cigarette
smoke derived from Oriental tobacco in the blend. Schumacher and
Vestal (1974) isolated 6-O-acetyl-2,3,4-tri-O-(+)-3-methylvaleryl)-
beta-D-gluco-pyranose from Oriental tobacco; this is responsible for
the 'dirty sock' aroma during combustion, which is characteristic of
Oriental tobacco.
Tobacco breeders have successfully increased sucrose esters in flue-
cured and burley and glucose esters in Oriental that have improved the
flavor of tobacco by increasing the concentration of free polar acids in
smoke (Smeeton, 1987). Acetic and formic acids are found in the
highest quantities of all the carboxylic acids in the tobacco and smoke
(Kallianos, 1976). These two compounds add acrid taste to smoke; at
the same time, these compounds alter smoke strength associated with
nicotine and ammonia produced from nitrogenous compounds. High
concentration of formic and acetic acids in tobacco also produce
irritation that is undesirable.
Aromatic acids are important flavor compounds found in Oriental and
flue-cured tobaccos. Phenylacetic acid, found in the leaf, imparts a
honey like sweet taste to smoke. There have been over a hundred or
more acid compounds identified from tobacco and tobacco smoke
(Kallianos, 1976). It is not possible to identify all the tastes and aroma
produced by the acids and acid derivatives in smoke; nor is it possible
to predict the effect of compounds without addition of amounts that
are probably greater than the amounts existing naturally. When adding
specific compounds to determine the organoleptic properties, it is
necessary to have a natural tobacco base before adding individual acid
compounds for smoke evaluation (Kallianos, 1976) in order to be able
to understand the sensory response of the added acid compounds.
The nonvolatile di-and tricarboxylic acids of tobacco make up nearly
90% of the weight of all the carboxylic acids produced in flue-cured
tobacco. The major nonvolatile carboxylic acids found in tobacco are
malic, citric, oxalic and malonic acids. Glycolic, succinic, fumaric and
pyruvic acids are found in lesser amounts. These compounds do not
exist as free acids but as salts in the leaf. The concentration found in
the leaf generally reflects the state of maturity of the leaf (Dawson,
1946), and this phenomenon is almost always reflected in the quality
of the leaf. For this reason, early tobacco chemists developed
formulae using chemical analysis to measure leaf quality. Studies by
Phillips and Bacot (1953) indicated that the quality of smoking
tobacco was directly proportional to reducing sugar and inversely
proportional to citric acid.
The poor smoking quality relationship with large quantities of citric
acid was later verified by Resnik, et al. (1957) comparing smoke from
cigarettes made from primings and other stalk positions of flue-cured
tobaccos. Other workers have suggested that the nonvolatile organic
acids do not contribute to aroma of smoke but improve its mellowness
(Abadallah, 1970). Leffingwell, et al. (1972) suggest that citric acid
adds to the body of smoke. Dawson (1946) indicates in his review that
nicotine is bound with citric acid and other nonvolatile acids in
tobacco. According to Shmuk (1953), the ratio of free nicotine to
bound nicotine in tobacco influences the quality of the smoke. For
instance, the poorer the taste quality the greater the content of free
nicotine. The addition of malic acid to tobacco has much impact on
smoke strength due to high nicotine malic acid mellowing the taste
and reducing the impact of high nicotine. The nonvolatile acids, found
as salts in tobacco, add to the total acidity of smoke, frequently
altering the balance (Abadallah, 1970).
i
Carotenoids
Eighteen carotenoids have been identified in green tobacco plants
(Weeks, 1986). These 18 compounds are degraded drastically from
2000 ppm in a vigorously growing green plant to less than 100 ppm
during senescense, harvesting, curing and storage. Breakdown of 40
carbon isoprenoid compounds in intact plants

 Page 310
occurs from lipoxygenases operating in the plant and in post-harvest
treatment by photo-oxygenation (Enzell, 1976). Enzell (1976)
suggested that photo-oxygenation occurs similar to biological
oxidation of unsaturated compounds which favor attack at 6,7-, 7,8-
and 9,10-double bonds in the polyene chain to produce 9-, 10-, 11-and
13-carbon compounds. Over 100 compounds with structures that are
in some way related to that of cyclic carotenoid degradation products
have been isolated from tobacco and tobacco smoke. These
compounds bear an oxygen molecule associated with the cyclohexane
ring or in the side chain and are usually water soluble with low vapor
pressure and capable of distilling from tobacco to smoke unchanged
(Enzell, 1976). The ionones, megastigmatrienones, damascones and
damascenones are considered to be the most important carotenoid
derivatives found in cigarette smoke. The ionones and damescenones
are primary aromatic compounds found in rose oil; therefore, these
compounds add floral and woody-like notes to the aroma and taste of
cigarette smoke (Roberts, 1988).
j
Phenolic Compounds
Phenolic compounds exist throughout the plant kingdom and are not
unique to tobacco. A number of simple phenolic compounds exist in
tobacco in small quantities but chlorogenic acid, rutin and scopletin
are the predominant phenolic compounds found in tobacco (Kallianos,
1976). Flue-cured tobacco contains higher concentrations of
chlorogenic acid, rutin and scopletin, than air-cured types (Penn &
Weybrew, 1958). The concentration of polyphenols varies with stalk
position and with flue-cured grades, and in that respect, polyphenols
are positively correlated with tobacco quality. Phenolic compounds
are easily oxidized to quinones and undergo rapid reactions with
amino acids, alkaloids and amines to produce colored pigments in
tobacco (Weeks, et al., 1993). Many of these colored pigments are
very high molecular weight resins and during pyrolysis produce few
compounds that contribute to flavor and aroma of smoke. Pyrolysis of
free chlorogenic acid and rutin produces simple phenols and
compounds which produce smokey-like aroma. Zane, et al. (1965)
demonstrated the formation of quinic acid-gama-lactone from
pyrolysis of chlorogenic acid in model systems. This lactone, with a
characteristic bitter taste, has been isolated from smoke but not from
tobacco (Zane, et al., 1965). At least 50 or more simple phenolic
compounds have been isolated from smoke, three times the number
that have been identified in the leaf (Kallianos, 1976). Some of the
simple phenolic compounds found in smoke derived from precursors
other than polyphenols have been implicated in bitter, smokey and
medicinal tastes of smoke. Although phenolic compounds such as
vanillin are added to tobacco as flavorants, as a general rule simple
phenolic compounds do not make important contributions to the
organoleptic properties of smoke (Penn & Weybrew, 1958). However,
some phenolic compounds can produce volatile compounds that add
flavor to smoke by degradation and rearrangement reactions that
occur during combustion (Kallianos, 1976).
k
Tobacco Alkaloids
Alkaloids, of which nicotine is the predominant compound, are
synonymous with tobacco (Enzell, 1976). There are volumes of
publications that address tobacco alkaloids. Harshness and smoke
strength obtained from air-cured tobaccos are due to the alkalinity
produced from nicotine (Leffingwell, 1976). Pyrolysis of nicotine at
high temperature produces substituted pyridines in tobacco smoke.
The substituted pyridines from pyrolysis of nicotine are responsible
for residual tobacco aroma following smoking (Jarboe & Rosene,
1961). Acetyl-pyridine, known for its characteristic tobaccolike
aroma, is a pyridine compound found in tobacco and smoke (Weeks,
et al., 1995).
Nornicotine, myosmine, anabasine and anatabine are other pyridine
alkaloids commonly found in tobacco. Negative effects of nornicotine
on smoking discourage the production of smoking tobacco with even
relatively small quantities of nornicotine (Roberts, 1988). Nornicotine
is formed from the enzymatic degradation of nicotine during
senescence and curing of tobacco. During pyrolysis, nornicotine
produces myosmine and substituted pyridine compounds that create
objectionable smoke flavor. Experiments with 14C nicotine and
nornicotine showed that nornicotine was a more efficient producer of
pyridine than nicotine (Enzell, 1976). The characteristic alkaline
aroma detected from pyrolysis of tobacco was associated with
nornicotine and nicotine eluted from a capillary GC column fixed to a
sniffing port and GC mass spectrometer during the pyrolysis of 'cherry
red' flue-cured tobacco that contains high quantities of nornicotine.
Myosmine obtained at the same time from the pyrolysis of 'cherry red'
tobacco produced a mousy like aroma (Weeks, et al., 1995). a,b-
Dipyridyl, a cigar-like aroma, has been isolated from the pyrolysis
products of cured and aged tobacco, a,b-Dipyridyl was previously
found in cured

 Page 311
tobacco following 14C labeling of anatabine (Enzell, 1976).
Summary
Tobacco is grown and manufactured for different consumer
satisfactions when compared to other agronomic crops grown for food
and fiber. However, this discussion has largely addressed constituents
of the tobacco leaf and substances produced from these constituents
during combustion and their influence on smoke. The purpose of this
monograph section was to bring to the attention of those individuals
interested in the influence of smoking, at least, some of the factors
that produce satisfaction as well as some of the components in
tobacco and smoke that produce feelings and responses that are less
satisfying. For this reason, there are many tobacco products available
for the smoker. Due to the complexity of the raw materials and the
combustion products, it is difficult for the tobacco leaf blender to
develop a product that is satisfactory for all customers.
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Chortyk, O.T., Severson, R.F. & Higman, H.C. (1975)
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Demole, E. & Dietrich, P. (1977) The chemistry of burley flavor. In:
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 Page 313

Chapter 9
Physical Properties of Leaf Tobacco
Y. Nakanishi
Japan Tobacco, Inc
Yokohama, Japan
Introduction
In this chapter the most important physical properties of the leaf, that
is, filling value, equilibrium moisture and burn rate, will be covered.
Strip yield and byproduct utilization are omitted as there are very few
recent studies dealing with these topics.
The first section of this chapter outlines filling value in terms of ways
to measure it and its affecting factors. In the second section,
equilibrium moisture in terms of the definitions and measuring
methods will be discussed and a recent examination from the
viewpoint of adsorption will be presented. Finally, burn rate will be
reviewed briefly in the third section.
Filling Values
Since the number of cigarettes with an appropriate hardness that can
be produced from a given weight of raw material directly affects
manufacturing cost, filling value is the most important of all the
physical properties of the leaf ('appropriate hardness' refers to the
hardness maintained before and during smoking that is acceptable to
the smoker). Filling value is an index for the number of cigarettes that
can be produced from a given weight of raw material.
Today, shifts in smokers' preferences away from full-flavor to mild
cigarettes and a changing smoking environment call for lower nicotine
and tar contents. The primary factor that determines these
characteristics is the cigarette burning rate, which is closely connected
to the amount of cut rag (shred) that fills a cigarette (Mulchi, 1982).
Thus, the filling value is more important than cost alone.
a
Measuring Methods
There are many measuring methods available to determine the filling
value depending on the compression method employed such as
triaxial, centrifugal, vibration and constant velocity and weight
(Artho, et al., 1963; Canon, 1976; Flesselles, 1971; Kobari, et al.,
1996; Kobata & Akinaga, 1956; Lorenz & Seehofe, 1968; Masuo, et
al., 1966; Masuo, et al., 1967; Voisey and Walker, 1969; Voisey &
Walker, 1970; Wakeham & Watts, 1976; Walker and Voisey, 1972).
Each of these approaches is designed to estimate the weight of cut rag
in a cigarette as accurately as possible, and the filling value is given
either as a measured mean specific volume of shreds or a reciprocal,
i.e. bulk density. The most widely used approach is to place a pre-
determined weight of cut rag into a cylindrical vessel to determine the
mean specific volume or bulk density when the flakes are compressed
under certain conditions. The two basic methods depending on
compression conditions are:
(1) constant velocity compression in which the cut rag is compressed
until the stress reaches a certain level, and
(2) constant weight compression in which a specified weight is
applied to the cut rag for a certain length of time.
The first method assumes that the fill density in a cigarette of standard
hardness has a compressive stress of 250 g/cm2 (Masuo & Shinozaki,
1972). The second method constant weight compression method is
currently the most common throughout the world. This typically
involves applying a 2 kg weight to an accurately measured 10 to 20 g
sample of cut rag in a 60 mm diameter cylindrical vessel for 30
seconds and measuring the mean specific volume of the compressed
flakes. It should be noted, however, that cigarette makers have not
agreed on standard conditions or on ways to take measurements. Most
companies use their own methods and conditions.
Saito and Abe (1977) suggested that there is a need to offset the effect
that friction has on the cut rag/vessel wall interface in order to obtain
filling values

 Page 314
independent of tobacco type because they found that compression
causes friction between the walls of the vessel and the flakes that is
specific to the tobacco type used. To minimize the effect of friction on
pressure measurement, Masuo, et al. (1967) suggested using constant
velocity compression that limits the pressure detection area to the
central quarter of the sample surface area to which pressure is applied.
b
Factors Affecting Filling Values
Factors that affect filling value may be divided into two major groups:
the characteristics of individual constituents comprising a cut rag
aggregate and the structure of the aggregate. Aggregate structure, the
second group of affecting factors, depends on how cut rag is packed
into a sample vessel using such compression methods as vibration,
constant velocity and constant weight.
Flesselles (1982) suggested density, elasticity, plasticity and
brittleness for the first group of factors that affect cut rag
characteristics. Saito (1974) also proposed the three factors of density,
elasticity and morphological characteristics (e.g. crimping, curling,
twisting, width and length). It seems quite clear that the major
characteristics of shred include the morphological ones proposed by
Saito plus thickness, all the characteristics of elastic solids (including
those that appear after buckling) and density. Filling value decreases
as temperature and moisture content rise since increased temperature
and moisture reduce the Young's modulus and facilitate buckling. It
appears difficult to derive an empirical formula that can estimate the
wide range of influences that all these characteristics would have on
the filling value due to difficulties of independently changing each of
these characteristics. However, Kato, et al. (1980) reported on an
experimental examination of morphological characteristics such as
crimping, curling and twisting using strips of paper and proposed that
their influences on the filling value can be expressed as the apparent
volume/displacement ratio which is defined as a ratio of the
rectangular parallelepiped circumscribing cut rag to the net volume of
this shred.
Quantifying the effects of the structure of a cut rag aggregate on the
filling value is even more difficult. To define an aggregate structure
requires a three-dimensional orientation for each flake, distribution of
voids (apertures) and their orientation. Goldring, et al. (1973) have
reported on the orientation of cut rag flakes in cigarettes. There are
numerous reports (Kalimes & Corte, 1960; Komori & Makishima,
1977a; Komori & Makishima, 1977b) in the field of textiles that have
studied the orientation of fibers based on statistical thermodynamics
but there are no examples, to our knowledge, of such a strategy
applied to cut rag tobacco. In the case of textiles, when considering
orientation it is sufficient only to take into account the direction of the
axis. In the case of cut rag tobacco, however, consideration must also
be given at least to length and width directionality. Therefore, from
both a theoretical and experimental perspective, it is a more
challenging task than with fibers. This area awaits investigation and
merits further studies.
Comparing the filling values of two representative tobacco types,
burley and flue-cured, we find that those for the burley are higher.
This is attributed to the fact that burley results in a higher Young's
modulus and buckling loads and lower apparent density than the flue-
cured tobaccos. The filling values for these tobaccos can be improved
by manipulating the aforementioned affecting factors. The most
innovative of these improvement techniques is R.J. Reynolds'
pioneering application of chlorofluorocarbons (CFC) to expand
tobacco tissue, an expansion process in which apparent density is
reduced. A variety of expansion processes have been developed and
commercialized based on a number of other agents besides CFC
(Kertsis & Sun, 1984; Kramer, 1991). Expanding shred reduces the
apparent density by more than 50%. However, the filling value could
only be improved by a factor of 1.5. This may be due to changes in
the shape of the cut rag induced by expansion processing, which in
turn change the structure of the aggregate, although details are not
clear yet.
Besides reducing the apparent density there are also a variety of
techniques proposed for improving the filling value by changing the
characteristics of cut rag or aggregate structure for which many
patents are pending from many cigarette companies. Of all these
techniques, only Japan Tobacco's crimped rolling sheet technology is
mentioned here as an example of practical technologies that change
the shape of cut rag (Ohashi, 1988).
Equilibrium Moisture
Equilibrium moisture content is the moisture content of a tobacco
(solid)water system in the state of adsorption equilibrium. Tobacco
moisture content is an extremely important measure of physical
properties closely linked to mechanical properties, burn rate and

 Page 315
taste. Naturally, considerable research has long been carried out in this area
(Wilson & Fuwa, 1922; Kamei & Suzuki, 1941b; Locklair, et al., 1957;
Bethmann, et al., 1961; Seehofer, et al., 1961; Höschen, 1965; Samejime,
al., 1978).
To measure equilibrium moisture content, dry weight must be established
first. However, since tobacco includes volatile constituents, the dry weight
will vary considerably according to the drying conditions. Over-drying will
change the constituents themselves while insufficient drying would result in
releasing the volatile components while taking measurements. In either case,
the measured moisture values will rarely be reproducible. Consequently, the
first requirement is to decide on reasonable drying conditions, a number of
which have been proposed and employed including vacuum drying, through
flow drying, as well as drying by heating (Wilson & Fuwa, 1922; Kamei &
Suzuki, 1941a; Locklair, et al., 1957; Samejima, et al., 1978; Nakanishi &
Kobari, 1988).
a
Measurement
There are three main ways to measure equilibrium moisture content. One
method takes measurements by a gas with a fixed relative humidity that
flows through a sample. Another method measures equilibrium moisture
under the water vapor environment after evacuating air. The third way uses
a desiccator and adjusts the humidity with a sulfuric acid solution or a
saturated salt solution and takes measurements under atmospheric pressure
(Kobari, et al., 1996). The disadvantages of the first method are the
difficulty of maintaining constant humidity for the long time required by
this method and preventing condensation is challenging. This makes it
impossible to determine moisture content in the high humidity ranges. In the
second method, measurements of moisture content can be taken over a
broad range of temperature and humidity conditions in an extremely brief
time. The third method cannot be used to measure moisture content in the
high humidity ranges because molds grow on the sample during the long
period of time required to obtain the measurements. Thus for plant materials
like tobacco, the second method is the most appropriate.
b
Equilibrium Moisture Content
Equilibrium moisture varies considerably according to tobacco type and
curing method (Locklair, et al., 1957; Samejima, et al., 1978). Samejima,
al. (1978) studied equilibrium moisture contents with flue-cured, Oriental,
burley and air-cured domestic tobaccos. The results of their studies showed
that equilibrium moisture declines in the following order when a relative
humidity is 40% or less: domestic tobaccos, burley, flue-cured, Oriental.
They reported that at 60% relative humidity or above the order was flue-
cured, Oriental, burley and domestic tobacco in descending order (Fig. 9.1).
Moreover, the hysteresis-different equilibrium moisture values in the
adsorption and desorption process, under the same conditions of
temperature and humidity, have rarely been reported for tobacco (Kamei &
Suzuki, 1941b; Nakanishi & Kobari, 1988).
 Fig. 9.1
Moisture sorption isotherms of various tobaccos at 293 K (from Samejima, et al., 1978).
c
Adsorption Isotherm Equation
The adsorption isotherm describes the relations between equilibrium
moisture and vapor pressure or the relative vapor pressure. The water
adsorption iso-

 Page 316
therm for tobacco forms a sigmoid and can be described well by the
BET equation in a relative humidity range of 0.17 to 0.5 (Samejima,
et al., 1978; Nakanishi & Kobari, 1988). Kamei and Suzuki (1941b)
employed a modified Langmuir isotherm equation for water which
includes a term of second degree of relative humidity in the
denominator than can be applied in a wide humidity range. Recent
attempts have applied the GAB (Anderson, 1946) and modified
Dubinin-Astakhov (DA) equations (Kameoka, 1995) to the isotherm
of water adsorption by plant materials and their processed products.
The GAB equation provides good agreement when the heat of
adsorption is smaller than the heat of condensation above the second
adsorption layer, namely when adsorption is caused by dissolving in
adsorbent (Yano, 1992). Nakanishi and Kobari measured the
adsorption isotherms of flue-cured and burley tobaccos to obtain their
equilibrium moisture values and applied them to the Clapeyron-
Clausius equation to obtain the changes in the isoteric heat of
adsorption in response to the changes in the coverage which is defined
by an 'amount adsorbed/amount adsorbed in monomolecular layer'
equation (Nakanishi & Kobari, 1988). The reported results showed
that the value of the adsorption heat for water was several times that
of the condensation heat when the coverage was less than one, but that
when coverage was more than one, it was almost equivalent to the
condensation heat and, except for an extremely low relative vapor
pressure, this indicated that the water adsorption was physical
adsorption. Thus, it is inappropriate to use the GAB equation. On the
other hand, the DA equation is a semi-empirical equation that
formalizes adsorption volume and the adsorption potential curve and
is applicable to a broad range of materials. Miyauchi, et al. (1995)
reported that they used the DA equation and were able to satisfactorily
describe the isotherm of water adsorption by tobacco in a relative
humidity range of 5 to 80%.
d
Pore-Size Distribution and Specific Surface Area
Since tobacco is a porous material, its equilibrium moisture is a close
correlation of pore size distribution and specific surface area. In most
cases, pore size distribution is measured using the mercury injection
method or the nitrogen adsorption/desorption method. Forcing
mercury into the pores of soft materials like tobacco deforms the
pores, and thus the first method may not provide the correct
distribution. Nitrogen adsorption/desorption also requires that the
sample first be dried. However, since drying dehydrates and
condenses the hydroxyl bases in cellulose, this causes plant materials
to shrink (Casey, 1983). Thus, Nakanishi and Kobari proposed the use
of the critical point drying method before using nitrogen
adsorption/desorption to measure pore size distribution and specific
surface area (Nakanishi & Kobari, 1988). This procedure prevents
pore contraction by soaking the material in an organic solvent, which
displaces water within the pores, replacing the organic solvent with
CO2, and finally drying the material at the critical temperature of
CO2. Using the resulting pore size distribution and specific surface
area, the amounts of capillary condensation and the multimolecular
layer adsorption were calculated, and adding them to give the
moisture content provided good agreement between calculated and
measured values. Exhibited in Fig. 9.2 are the cumulative curves of
pore volume obtained for flue-cured and burley tobaccos. As the
figure shows, the pore volume of flue-cured tobacco rises with
moisture content that changes the structure. On the other hand, that of
burley tobacco remains almost unchanged with moisture content.
Thus, the difference in water adsorption characteristics between
burley and flue-cured tobaccos is well described in terms of pore size
distribution.
e
Effects of Flavoring on Equilibrium Moisture
To improve taste, a variety of flavorings are added to tobacco.
Generally, adding flavorings and other additives changes equilibrium
moisture content. Nakanishi and Kobari (1989) performed theoretical
and experimental studies on the binary adsorption equilibria of water
and ethanol as reported below. When ethanol is adsorbed in the
multimolecular layer and/ or the capillary condensed phase of water,
the mixing of the water and the ethanol reduces the chemical
potential. This serves to increase the amount of adsorption of both
adsorbates. On the other hand, the water-ethanol surface tension
sharply declines in the capillary condensed phase as ethanol
concentration increases and the capillary condensation radius becomes
smaller. This tends to reduce the amount of these two substances that
is adsorbed. In the water-ethanol adsorption equilibrium, these two
opposite effects offset each other so that the tobacco equili-

 Page 317

Fig. 9.2
Variations of the cumulative curve of pore volume
for (a) flue-cured and (b) burley tobacco with
humidity. The samples (1) were desiccated by
vacuum drying at 353K, while the samples (2,3
and 4) were prepared at the relative humidities
of 30, 60 and 80%, respectively and then
desiccated bycritical point drying after
dehydration by aqueous ethanol solution (from
Nakanishi & Kobari, 1989 and Nakanishi &
Kobari, 1988.
brium moisture scarcely changes. Miyauchi, et al. (1996a) examined
this using several model fragrances (that are not much soluble in
water) which experimentally supported the above arguments. On the
other hand, it has been reported that with moisture retaining
polyhydric alcohols, equilibrium moisture increases with the amount
added (Samejima, et al., 1978; Miyauchi, et al., 1996b). This may be
explained by the fact that the surface tension of polyhydric alcohol
solutions is less dependent on concentration than is the case with
ethanol solutions and this prevents the capillary condensation radius
from becoming appreciably smaller when water and polyhydric
alcohols are mixed.
Burn Rate
The burning rate of cigarettes is controlled mainly by the wrap paper,
more specifically its permeability and the amount of burn-control
additives in it. Filter ventilation also contributes to controlling the
burning rate (Durocher, 1984). Thus, the raw material is not
considered particularly important in determining the burning rate.
Today, however, changes in smokers' preferences and the smoking
environment call for lower nicotine and tar. To produce cigarettes with
low nicotine and low tar while retaining sufficient smoke, it would be
effective to reduce the number of puffs. This has refocused attention
on the combustibility of the raw material itself. This chapter does not
cover the effects of wrap paper permeability, burn-control additives,
cigarette circumference or filter ventilation on the burning rate since
these topics are covered in detail in Chapter 12 Smoke Chemistry.
a
Measurement
Cigarette burning rate is expressed by either the number of puffs or
the static burning rate under standard smoking conditions. Since there
is a high correlation between the number of puffs and the static
burning rate (Resnik, et al., 1977), the static burning rate is widely
employed because of its simplicity. Please refer to International
Standards Organization (ISO) 3612 for the static burning rate
measuring method. The static burning rate is described in terms of
length burned per unit of time (mm/min) which is the linear burning
rate or in terms of quantity, that is, the mass burning rate (g/min).
b
Burn Rates
As shown in Table 9.1 (Muramatsu, 1981), when tobacco types are
compared, burley exhibits higher linear burning rate and mass burning
rate values than flue-cured tobacco. The burning rate of the lower
stalk is also greater than that of the upper stalk. To examine the
burning characteristics of expanded tobacco, Ihrig, et al. (1986) made
cigarettes with different distribution ratios of nonexpanded and
expanded tobacco then measured their burning rates. The results are
shown in Table 9.2. In comparison with nonexpanded tobacco, the
expanded cut rag exhibited a higher burn rate. However, the
comparison did not reveal large differences in the mass burning rate.

 Page 318
Table 9.1 Burning rates of flue-cured and burley tobaccos.
Linear Mass Peak
TobaccoStalk burning rate burning rate combustion
type position Density (mm/min) (mg/min) temperatures
(g/cm-3) (°C)
Flue- Cutter- 0.258 3.21 41.6 812
cured 2
Lea-l 0.281 2.66 37.5 794
Leaf-2 0.304 2.48 36.6 805
Burley Cutter- 0.219 4.55 50.1 807
2
Leaf-1 0.256 3.80 48.9 806
Leaf-2 0.256 3.41 47.6 802
Source: Muramatsu (1981).

Table 9.2 Comparison of burning rates of nonexpanded and expanded

tobaccos.
Density Linear Mass burning
Blend CircumferenceWeight (g cm- burning rate rate (mg min-
(mm) (mg) 3) (mm min-1) 1)
100% 24.8 846 0.275 3.79 50.9
Nonexpanded
tobacco
50%
Nonexpanded
50% 24.8 622 0.201 4.93 48.7
Expanded
tobacco
100% 21.8 667 0.280 4.20 44.5
Nonexpanded
tobacco
100% 21.8 363 0.153 7.62 4.39
Expanded
tobacco
Source: Ihrig, et al. (1986).
The burning rates of raw materials varied according to their
constituents. Those that have the greatest bearing on burning rates are
potassium and chloride (Akehurst, 1981). In order to study the effect
that the concentrations of these constituents have on burning rates,
Chaplin and Miner (1980) and Mulchi (1982) controlled amounts of
potassium and chloride in soil and cultivated tobaccos under these
environments, respectively. The reported results showed that higher
levels of potassium concentrations increased the burning rate, while
higher concentrations of chloride reduced it. Thus, potassium
contributes favorably to combustibility while chlorine impedes it.
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(Dai 1 Hou) (in Japanese). Kogyo Kagaku Zasshi, 44, 1269.
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equilibrium moisture content of cell wall materials. Kagaku Kogaku
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hardness of cigarettes. Beitr. Tabakforsch., 4(7), 293300.
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Tokyo.

 Page 320

Chapter 10
Marketing, Processing and Storage
10A
Tobacco Marketing Systems
John S. Campbell
John S. Campbell Ltd
Wilson, North Carolina, USA
Introduction
Tobacco is cultivated for the purpose of providing a raw material,
which, after manufacture, satisfies the requirements and desires of its
ultimate consumer, principally the smoker, wherever he lives.
Therefore, the grower should aim to produce and market leaf which is
wanted by the buyer, leaf processor and, manufacturer, in whatever
area he operates, throughout the world. The grower and the buyer are
also dependent on each other for the provision of an acceptable raw
material at a reasonable price. Both, however, require an adequate
profit margin to finance their operations.
The method of growing, harvesting and curing the many tobacco
types (flue-cured, burley, light and dark-air cured, Oriental and semi-
Oriental, dark-fire) varies between countries and areas, thus providing
a range of acceptable leaf, with differing physical attributes,
chemistry, flavor and aroma to meet various smoking demands. Soil
types and environments, notably climate, which change from year to
year, together with the agronomic practices imposed on them in the
field, affect the many types grown. Finally, harvesting and curing
practices may levy further variables as they relate to the presentation
of the leaf at the market place.
System of Production and Marketing
Marketing of tobacco types by growers may be by an auction system
or by contract. In either case, it is desirable to balance production by
some means, e.g. voluntary or strict farm quotas, to ensure cured leaf
sales in domestic/export markets. Accurate prediction of leaf demands
(domestic/export) and future yields, which can change due to varying
weather conditions as with all crops, make this difficult. Nevertheless,
any method to achieve long-term viability of a crop, say for 5 years,
provides stability to the grower with his need to finance and purchase
equipment. The Flue-Cured Tobacco Growers' Marketing Board in
Ontario, Canada has been particularly cognizant of this approach in
future development and allocation of quota. The systems of marketing
will be described in the following sections.
a
Auction
USA, Canada, Zimbabwe, Malawi and India use the auction system;
principally for flue-cured and burley types, with the flue-cured
growers generally producing more tobacco (10 to 200 ha), whereas,
more often burley growers produce less tobacco (1 to 10 ha).
Production levels are voluntary in Zimbabwe and Malawi, though
annual recommendations are made by grower organizations. Leaf
quotas, based on predicted domestic/export demands, current leaf
stocks held by the growers and past yield/ha achievements, are
allocated to growers annually in the USA by the Secretary, United
States Department of Agriculture for all types, in Canada by the
FCTGMB and in India by the Tobacco Board.
There are currently 53 flue-cured auction markets and 42 burley
markets (1998) in the USA.; 2 flue-cured in Ontario, Canada; 1 flue-
cured, 2 burley in Zimbabwe; 1 flue-cured, 2 burley in Malawi and 28
flue-cured in India. The numbers of markets will change over time.


 Page 321
b
Allocation
In Australia major changes have been made since 1995. There are two
sales floors: in Mareeba, Queensland and Myrtleford, Victoria. There
are no auctioneers. Growers' lots are lined up in rows, as in normal
auctions. Bales are allocated to each buyer based on the share of
orders generated from separately negotiated contracts (three
manufacturers) and the ticket is appropriately marked. Two appraisers,
employed by the growers, grade each bale in company with a buyer
from each manufacturer. If grade agreement is not reached, further
arbitration may be required (small percentage). Prices for each grade
have been negotiated prior to this allocation procedure on the floor.
The appropriate figure is written on the ticket (Australian Tobacco
Marketing Advisory Committee, 1995).
c
Contract
In most tobacco growing countries in the world where small growers
(0.5 to 3 ha.) are in the majority, e.g. Europe, Africa, Asia, Central
and South America, tobacco is grown and marketed on a contract
basis. There are a number of variations on how this is carried out
which will be discussed.
Private Tobacco Manufacturing Companies
These organizations within countries, as in Brazil, Argentina, Chile,
Guatemala, Kenya, Uganda, Nigeria, Zaire, Ghana, Pakistan, Sri
Lanka, Bangladesh, Mexico and Venezuela, contract with farmers to
grow tobacco.
The tobacco is produced and marketed under a contractual, signed
agreement between growers and the company prior to the growing
season. These companies may provide certain financial inputs, seeds
and/or plants, fertilizers, crop protection agents, curing facilities and
fuel, together with technical agronomic advice during the growing
season. Such an arrangement enables the manufacturers to obtain
qualitative and quantitative characteristics desired in their products.
They may even select or advise on the most desirable areas and soils
on which the tobacco should be grown. These contracts will vary
slightly from country to country. The grower in most instances will
contract his entire crop to one company, but in some places, to one or
more. In certain countries, e.g. Kenya, Brazil, he may be required to
cultivate trees on his farm to provide wood for curing his flue-cured
tobacco (Campbell, 1994).
Tobacco Leaf Dealers/Merchants
These groups as in Brazil, Mexico, Spain, Thailand, Italy, Greece,
Hungary and the Philippines, or grower cooperatives as in Argentina,
South Africa, France and Italy, contract for tobacco production. In
Turkey all contracts are made with the government monopoly, but
growers are also allowed to sell to dealers.
Their contract system is very similar to that of domestic
manufacturing companies. Such contracts may vary slightly from
country to country in the amount of financial inputs and technical
assistance. The grower may contract to one or more dealers or
cooperatives. The leaf dealers may have primary processing facilities,
in which case the leaf purchased is threshed and may be sold to
domestic manufacturers or for export. In Argentina the leaf
purchasing and primary processing by the cooperatives on behalf of
leaf dealers are closely supervised by both parties to ensure that both
buying and threshing specifications are met.
Financial repayment problems sometimes arise with growers when the
climatic conditions during the growing season, e.g. hail, torrential
rains or floods, or drought, can markedly reduce yields or leaf quality.
Despite this, companies or cooperatives will try to alleviate such a
problem and, at the same time, attempt to retain good growers from
year to year.
Government Tobacco Manufacturing Monopolies
These are found primarily within free market countries, such as Japan,
Thailand, Taiwan, the Philippines, Turkey, Italy, Portugal, South
Korea and Spain. Some of these monopolies are in the process of
change to private ownership, notably in Western Europe and Japan.
Most control tobacco output on an annual basis and contract with
individual growers under similar marketing systems to those described
above for manufacturing companies and leaf dealers. China follows a
similar procedure.
Since 1992, the European Union, composed of countries whose
monopolies, leaf dealers or cooperatives contract the production of
leaf, establishes quotas for each member country and for each of five
tobacco groups (flue-cured, light air-cured, dark air-cured, fire-cured
and sun-cured types) and three Greek Oriental varieties (European
Union, 1992). This quota system was reviewed in 1997.
The political breakdown of communist Russia and its associated
Comecon countries has resulted in major changes. Leaf production
was often carried out under

 Page 322
contract through collectives/communes, not individual farms. Indeed,
this is generally still the case in Moldova, Uzbekistan and Bulgaria.
Collective contracts were, and for the most part still are, with
government or other manufacturing monopolies, such as in Russia and
members of the Commonwealth of Independent States (CIS),
Bulgaria, Albania, Romania and Poland. In some republics
independent merchants are now being set up. In recent years, leaf
production has diminished markedly.
Major changes in production and marketing in these areas are
anticipated, as they change from centrally planned systems to market
economies. These will depend on future government policies, capital
availability, notably for inputs, changes to private ownership of land,
influx of multinational take-overs and privatization, consumer brand
and blend changes, leading to the production of different tobacco
types, and a major change in personal attitudes. It has been stated that
'Bulgaria is in a transitional period, undergoing a change of system
and structure' (John, 1994). This can perhaps be applied to all these
countries in relation to grower production and marketing. The vital
need to increase leaf production to meet future usability requirements
of the buyers/manufacturers in these areas has been expressed by
Barton (1995), not only to avoid the use of hard currency for leaf
imports, but also to improve local economies and provide profitable
employment.
Flue-Cured Green Leaf to Independent Curing Barn
In this growing and marketing system, the mature green leaf is picked
by the grower, who delivers it to independent curers, such as in
Malaysia or Thailand. The National Tobacco Board (NTB) in
Malaysia and the Thailand Tobacco Monopoly (TTM) set annual
quotas, which are divided up amongst the curers and thence to the
growers. In Malaysia, the green leaf is purchased by the curers, by
grades (17 are described) and prices set by the NTB, then cured in
their barns and in turn sold to leaf dealers or tobacco manufacturers.
The reluctance by the tobacco growers and curers to adopt modern
methods of planting and curing has contributed to poor quality
tobacco. Furthermore the curers act as 'middlemen', adding an extra
cost (Anon, 1991). In both countries, it would be desirable if there
were a direct contract between grower/dealer/manufacturer. In Fiji, the
manufacturers purchase green leaf from the grower and cure it
themselves (Akehurst, 1981). In Jamaica, there is a similar procedure,
but after curing in company barns, the grower grades his own leaf and
is paid accordingly.
Grading/Sorting by the Grower Prior to Marketing
Grading is the sorting of leaves to form a uniform pile, which can be
classified, as a lot, according to a specification. Since leaf chemistry,
smoking and physical characteristics correlate with plant position and
are very important to the buyer, grading by plant position and maturity
(ripeness) is highly desirable (Weeks, 1992). In well grown flue-cured
and air-cured tobaccos, leaves vary in length, width and body
(thickness), which helps to subjectively designate their separation,
together with color, texture and degree of damage, blemish or
spotting.
Since fire-cured tobaccos are generally topped lower in the field,
leading to less variation in shape and length of leaves, and Oriental
tobaccos have smaller and less variable length of leaves, such
subjective characteristics (notably stalk position), while still very
important, are perhaps less easy to define. Thus separation at harvest,
curing and in subsequent storage becomes essential. Obviously the
more uniform the origin of the particular lot (affected by method of
growing, harvesting and curing) the fewer the grades required and the
easier will be the sorting for all tobacco types.
It is important for the grower to store and market cured leaf at a
moisture content which will inhibit the development of fungal molds
or other damage to it. Moldy tobacco will not be purchased, since it
causes an off taste in manufactured products. The ideal moisture
content of cured leaf in most countries is 16 to 18%. Specifications
may vary from country to country. Thus, the European Union has
defined these specifications for all tobacco types (European Union,
1992). In India this figure is much lower in order to reduce spoilage
because of high humidities in transport. It must be borne in mind that
the buyer will avoid high moisture tobacco at point of sale, since he is
paying a higher price than necessary because of increased weight.
It is very important for the grower, when marketing his tobacco, to
avoid including foreign matter (string, grass, paper, plastic, hessian
fluff, rubber bands, grease or oil), parts of stalks or suckers, scraps or
sand, or any other organic material, which demands removal prior to
manufacture (hence an added cost) or which detracts from, or mars,
the smoking quality of the manufactured product.

 Page 323
The principal aim of grading or sorting by the grower is to provide a
salable product with a good appearance which the buyer will desire
and for which he is prepared to pay a good price.
Market Presentation
Flue-cured tobacco is hand or machine harvested, generally at least
three times or more, and cured in conventional or bulk barns. Most
conventionally cured leaf is carefully hand sorted and grower graded
by plant position and color. It may be tied in hands e.g. Brazil, Kenya,
Malawi, China, Zimbabwe and Tanzania, or straight laid loose, e.g.
Canada, Argentina, Mexico and Australia. The tobacco may then be
baled and covered with hessian sheets, e.g. Kenya, Nigeria, Tanzania
and Australia, or in open bales tied with string, e.g. Brazil, Ghana,
Mexico and Argentina. Variations include coveting with bituminized
paper (inner lining) and hessian (outer lining), e.g. Zimbabwe,
Malawi; covering bales with brown paper, e.g. Canada, or covering
with untied leaf between two pieces of hessian and tying with Bengal
twine, e.g. India. Such parcels may be collected by the contractor, e.g.
Kenya, or grower delivered to the auction floor or a local buying
station, e.g. India, or 'curer' delivered to a collective selling point, e.g.
Malaysia.
Bulk-cured tobacco may be placed, with little sorting, as loose leaf not
necessarily straight laid, in hessian sheets, hessian wrapped or
between baling boards, and delivered to the market, e.g. USA and
Western Europe. Bulk cured tobacco in Japan is placed in cotton bags
which, like hessian, are reusable (Japan Tobacco, Inc, 1995, personal
communication).
The weight of the sheet or bale varies among countries: up to 340 kg
USA.; 120 kg Zimbabwe, India; 100 kg Australia; 80 kg Malawi; 60
kg Mexico; 65 kg South Africa, Malaysia; 48 to 50 kg Greece; 40 kg
Japan; 25 to 50 kg Western Europe; 25 kg Hungary, Taiwan. For ease
of presentation on the market floor in Canada, up to 35 bales, of the
same classification and weighing 25 kg each, are placed on wooden
pallets. Research in the USA is underway to determine a potential
package modification for flue-cured tobacco. In 1998, approximately
13% of the USA flue-cured crop was sold in large farm bales (340 kg)
rather than sheets.
Burley tobacco may be stalk-cut, e.g. USA., Argentina, Brazil,
Mexico (40% of crop) and Spain, and cured in appropriately sized
barns, or hand primed and air-cured, e.g. Mexico (60% of crop), Sri
Lanka and the Philippines, or five to eight of the bottom leaves are
hand-primed and the remainder stalk-cut, e.g. Guatemala, Thailand,
Japan and Malawi. After curing, stalk-cut leaves are removed from the
stalk and kept separate by plant position and color. Primed leaves are
sorted and graded in a similar manner. The leaf may be tied in hands,
e.g. Brazil, Sri Lanka, or straight laid, e.g. Argentina, Mexico, Japan,
USA, and placed in uncovered uniform size bales weighing 45 kg,
tied with string, e.g. USA, Mexico, Brazil, Argentina, or in hessian
covered bales, weighing 60 to 80 kg, e.g. Guatemala, Zimbabwe,
Malawi, or in woven bags weighing 40 kg, with good ventilation, e.g.
Japan.
Oriental tobacco plants are hand harvested three to four times or more,
and each leaf is hand threaded onto string prior to sun-curing, e.g.
Turkey, Greece, Bulgaria and Thailand. After completion of curing, it
is baled on the string and stored until the following year before
marketing, often in the grower's own home, e.g. Turkey, Greece and
Bulgaria. In Thailand it is marketed in the same growing year.
The stringed tobacco is placed in hessian bales, with the top side open,
in bales of up to 55 kg (Izmir), up to 25 kg (Samsun), 22 to 28 kg
Greece, prior to sale on the farm, or in small bales of 10 to 15 kg,
delivered to buying stations, e.g. Thailand. Representatives of the
Turkish Monopoly (TEKEL) or National Tobacco Board (Greece) and
leaf dealers/merchants assess the tobacco grade make-up and purchase
it, generally the whole crop. The leaf is delivered to an appropriate
warehouse where it is manipulated (blended) and repackaged in
hessian covered bales prior to fermentation. Growers in CIS countries
normally remove the string before baling prior to sale.
Some Oriental tobacco leaf, often of a better quality, may be
destringed and bundled in 20 to 30 straight laid, loose leaf lots or
'pastals' and thence into bales. Baling, either with strings or as pastals,
is common in Bulgaria and in the classic Basma districts in northern
Greece.
It must be noted that Oriental or semi-Oriental leaf is fermented. The
fermentation is mild compared with that of dark air-cured tobacco and
is natural in Greece and Turkey and artificial in Bulgaria and the CIS.
No threshing, as occurs in flue-cured, burley or air-cured tobaccos, is
carried out. Such leaf is blended in whole form in the cigarette
manufacturing process.
Classification
The original purpose of classification of grower lots was to supply the
buyer with better selected, higher quality leaf and the grower with an
incentive to provide this as a means of obtaining higher prices
(Campbell, 1993).

 Page 324
However, the older classification systems were developed for
manufactured products consumed in much smaller amounts today.
Cigarettes now represent 90 to 95% of the world's consumption of
tobacco manufactured products. There are different categories of
cigarettes, namely Virginia, dark-air, Oriental or blended, composed
of various tobacco types in different countries. Thus, present-day
classification systems must relate to what is wanted in the market
place and encourage growers to produce such styles of leaf for the
final consumer.
Grower lots, either at auction or on contract, are classified by
competent graders/classifiers into official standard grades on which a
price may be bid by the buyer or which, in some countries, is
previously defined. Such classifications, which are modified from
time to time, differ from country to country but are generally the
result of consultation between appropriate Government departments,
grower cooperatives or associations and leaf dealers/merchants or
tobacco manufacturers. The final text of such classifications is often
published by appropriate government bodies.
a
Auction
The first government classification system was developed in 1924 in
the USA. Today in the USA, fluecured, burley and fire-cured tobacco
types have their own systems (Campbell, 1993). The quality
classification for fluecured (1 to 6, with 1 being choice quality) and
burley (1 to 5, with 1 being choice quality) describes the looks of the
cured leaf and identifies each lot or pile by stalk position, color and an
estimate of total quality based on 10 elements or characteristics:
maturity, leaf structure, body, oil, color intensity, width, length,
uniformity, injury and waste tolerance (USDA, 1986, 1989, 1990).
Fluecured has 153 grades and burley 113.
This system has served as the basis of the tobacco leaf grade
descriptive structure for many tobacco producing countries in the
world. Indeed, USDA personnel have trained, and continue to train,
graders/classifiers in some countries. The actual system finally
developed in any country must however, be of a practical size,
particularly relating to the grower's size of crop, but at the same time,
as stated above, encouraging them to produce what is desired, at any
one time, in the market place by the buyer (Campbell, 1993). They
may take into consideration the requirements of the domestic, as well
as the export leaf buyer/manufacturer.
In Zimbabwe, the Tobacco Marketing Board has developed a
fluecured classification system based on group (principally stalk
position), quality, color and style (principally based on texture and
maturity), and extra factors (generally emphasizing poor quality).
There are about 130 grades depending on each year's crop quality
(Tobacco Marketing Board, 1994a). Since a very high percentage of
this tobacco is exported, considerable emphasis has been placed on
providing the overseas buyer with a high quality leaf by plant position
and darker colors, ripe in maturity, well sorted and uniform. Burley
(grade number about 80) follows a similar approach (Tobacco
Marketing Board, 1994a). Though the grade is written on the lot
ticket, the buyer takes little notice of it. The collated information of
grades/prices provides useful information as the market sales progress
day by day, for both the grower and the buyer.
In Malawi, the Tobacco Control Commission has developed a
somewhat similar classification system with 203 grades for fluecured
tobacco (Malawi Tobacco Control Commission, 1994). Emphasis
again, as in Zimbabwe, is on tobacco exports.
In Ontario, Canada, the FCTGMB, since 1987 has encouraged
growers to concentrate on improving maturity levels, to produce leaf
of orange or mahogany colors, rather than lemon, and to improve
flavor and smoking characteristics, for both domestic and export use.
Sorting by plant position is emphasized. The classification system
follows a similar pattern to those discussed above, with 191 grades
(Government of Ontario Canada, 1989).
Indian tobacco auctions started in 1984/85. The Government Tobacco
Board provides assistance to growers in grading and sorting prior to
sale. Over the years, this has improved (Chandramouli, 1988). An
ideal classification system based on plant position, quality (including
blemish), color and maturity has been described (Imperial Tobacco
Ltd, 1993). In the northern light soils of Andhra Pradesh and
Karnataka, the classification of 63 grades emphasizes plant position.
In the black cotton soils of Andhra Pradesh, the 11 grades concentrate
on color and blemish (Imperial Tobacco Ltd, 1995, personal
communication).
b
Allocation
Australia has developed a grade structure comprising 84 grades with
particular reference to plant position, color, quality and with special
factors for 'common' tobacco. It is considered that maturity is
insufficiently recognized (Australian Tobacco Marketing Advisory
Committee, 1995).

 Page 325
C
Contract
The classification system of tobacco types grown under contract is
generally less detailed and the number of grades reduced, simply
because the majority of growers produce relatively small quantities of
leaf. However, they include similar descriptions to those under the
classification section. Numbers will vary from country to country.
Brazil has 48 market grades for fluecured and 24 for burley (Brazil
Governo do Estado do Rio Grande do Sul, Secretaria da Agriculture e
Abastecimento, 1994); Argentina 46 fluecured and 24 burley grades
(Nobleza-Piccardo, 1995); South Africa 127 for fluecured (Jordaan,
1995, personal communication); Mexico 13 to 14 each for fluecured
and burley (Association Rural de Interes Collectivo de Productores de
Tabaco, 1994); the Philippines 10 fluecured (Philippines National
Tobacco Administration, 1994); Kenya 17 fluecured (Government of
Kenya, 1994) and Taiwan 7 fluecured.
Systems change to some degree over time, some grades are dropped
while others are added. In some instances, countries try to improve
their standards to gain more international recognition of their
unmanufactured leaf and manufactured products. Thus, in the People's
Republic of China, as from 1992, the China National Tobacco
Company is encouraging many growers to produce cured leaf of a
riper, more flavorful style with a wide range of chemistry, not only
available for inclusion in better quality domestic cigarettes but also
unmanufactured leaf for export (Ridgeway, 1992). There is one
grading system comprising of 40 grades, for both domestic and export
leaf types.
The subjective characteristics described in tobacco grades present at
auction or on contract may be useful to the buyer, e.g. uniformity of
plant position and color; however, many of them are not necessarily
synonymous with the descriptive method of the customers' grades.
The buyer does not base a choice on such a grade allocation, but
rather his judgments are related to subsequent green leaf processing
and manufacturing demands. Each customer, leaf dealer or
manufacturer has his own grading system which differ in degree
(Campbell, 1991). Thus, it is customary to regrade all purchases
received from the auction floors or from contract growers into buyer
grades desired by customers.
Marketing Regulations
In most countries growing tobacco, there are Government Tobacco
Boards, who legislate, control and enforce rules and regulations that
relate to the total industry. Unless they are a part of a complete
government monopoly, such Boards work in close consultation and
cooperation with growers' organizations and cooperatives, with
private tobacco manufacturers and with leaf dealers/merchants.
Strict and detailed regulations have been laid down for the
implementation of leaf sales by auction. These relate to such items as
grower registration, grower crop estimates, delivery quotas and
bookings (date and time often a number of occasions, as the market
progresses), method of leaf presentation and type of packaging,
weighing procedures and ticketing, grower use of crop protection
agents, grower refusal of sale and resubmission for resale, system of
payment after sale and procedures for appeal and arbitration.
Regulations governing building codes, notably natural lighting
through the roofs of warehouses in the USA or artificial lighting in
Canada, are important for accurate leaf classification and for buyers at
sale time.
Marketing regulations are promulgated by the USDA Agricultural
Marketing Service in the USA (USDA, AMS, 1994), by the
Government of Ontario, Canada (Government of Ontario Canada,
1989), the Tobacco Marketing Board, Zimbabwe (1994b), The
Tobacco Control Commission, Malawi (1994) and the Tobacco Board,
India. Selling procedures have been adopted in Australia (Australian
Tobacco Marketing Advisory Committee, 1995).
Each tobacco pile or bale is officially classified before sale, and the
buyers have time to inspect such tobacco prior to opening of sales. As
a cost reduction procedure, Zimbabwe only classifies every fifth bale.
Purchases are made very fast indeed with an auctioneer, a starter,
ticket writers and a set of buyers. The sale price and buyer's name are
written on the ticket, and the grower has the right of rejection. The
only exception is the Dutch Clock system in Canada (Menzie &
Marshall, 1981). Leaf bales are displayed for usability assessment by
the buyer on the exchange (market) floor. Purchases are made by a
counterpart in a separate room. On the clock, prices start high and
move to lower levels. Buyers bid by stopping the clock for their
appropriate lot. Again, the grower has the right of rejection within 30
minutes of sale.
The marketing regulations for all contract systems are much
simplified and are governed by the requirements of the contractor. In
some instances simple regulations made by the government, e.g.
Republic of South Africa Marketing Act, 1968 (Jordaan, 1995) and
the Kenya Legal Notice of 1994 (Government of Kenya, 1994), will
define similar items, as under auc-

 Page 326
tion above. While it is not always desirable for government
intervention, this may be necessary to ensure the grower sells to one
company and to reduce unethical competition.
Leaf Pricing and Payment
It must be noted that there is a very wide range of sale prices of
tobacco products, notably cigarettes, on the international scene, from
the hand-made, green cut and dried leaf (kerf) in cigarettes in East
Java and Vietnam, to the bidi cigarette so widely smoked in India, to
the cheap dark air/Oriental blends of Eastern Europe and CIS, to hand
rolled cigarettes and finally to the more sophisticated cigarettes of
Western Europe, Japan, Australia, New Zealand and North America.
Even where there is a higher standard of living and greater cash
availability, the 'discount' or generic cigarette is much in vogue. Each
of all these 'brands' of cigarettes requires leaf and the price and
usability of that leaf determines where it is blended and sold. Thus,
the grower is producing leaf for a specific market, either for domestic
use or for export.
Internationally, growers are very much affected by the world supply
and demand situation. Buyers are well informed of the daily
availability of tobacco from all sources, so that they can switch
purchases to cheaper leaf sources if they are considered suitable to
include in their brands. It is, therefore, highly desirable to build up a
reputation for producing a particular usability at the right price.
Growers and buyers, in association, have to identify their markets and
develop sales.
a
Pricing
The pricing structure under all marketing systems can help to increase
or decrease production. It can help to manipulate the quality of leaf
desired by the company, organization or monopoly buying or
contracting the leaf. Under a free auction system, e.g. Zimbabwe, the
grower can note very quickly what the buyer wants and change his
agronomic practices the following year. This has happened in Brazil
(Campbell, 1993) and has occurred in China (Ridge-way, 1992) for
leaf exports. Extra bonuses can be paid at the end of the season to
those growers producing a large percentage of good quality leaf.
Everywhere, prices paid to growers must reflect their cost of
production, plus a management profit, to ensure continuity in tobacco
growing.
Auction
In the USA there is a support price for each grade. If that price is not
exceeded by the buyer, the tobacco goes under loan into a cooporative
stocks pool (owned by the growers) until such time as it can be sold.
As a condition for eligibility for price support, growers must comply
with certain regulations, defined by the USDA, AMS (1994). The
high level of these support prices has not encouraged the production
and sale of leaf for export, therefore leaf exports have declined over
the years. On the other hand, minimum support prices, annually
agreed between growers and domestic buyers, coupled with grower
emphasis on plant position, color and maturity, incorporated into 'A'
grades have encouraged the production of leaf for export in Ontario,
Canada. Basically any support price should be a safety net just to
cover cost of production.
Allocation
Australian manufacturers negotiate separate contracts for leaf
quantities and prices with grower cooperatives or State Boards. Each
manufacturer has agreed to purchase leaf on a run-of-crop basis, as
from 1995, for a period of around 5 years at levels that are broadly
equivalent to a 57% domestic usage level in cigarettes. Prices are
expected to marginally decline over the next 3 years. Even so, such
leaf prices are significantly above those of the world market.
Contract
In many countries, prices for each grade are set annually by
Government Boards after consultation and agreement with growers
and buyers, e.g. the Philippines, South Africa, often before the
growing season opens and contracts are agreed and signed. Where
private companies are involved directly with growers, e.g. Guatemala
and many countries in Africa, or government monopolies, e.g.
Taiwan, China and Japan, they will set the prices.
In Brazil, prices are established in advance by the growers'
organization (AFUBRA) and the tobacco companies' organization
(SINDIFUMO) without government intervention. Buying companies
may change these prices up and down depending on the rate of
inflation between the period of agreement and actual purchase.
In Argentina, prices are negotiated in conjunction with growers,
cooperatives, leaf dealers, government

 Page 327
and manufacturers. A price is set on grade B1F and all the other grade
prices are calculated as a percentage of this price. Government
collects a tax on the sale of cigarettes and part of this money is given
to the growers as a subsidy (FONDO) depending on their grade
turnout, rewarding the better grades more and the poorer grades less.
There is a similar arrangement in Mexico, as in Argentina, whereby
the growers' organization (ARIC) and the two tobacco manufacturers
and two leaf dealers agree to an average price, based on what buyers
anticipate in their grade make-up of their contracts. Privatization is
underway in many countries.
A complicated system of pricing is contained in the European Union
(EU), Common Agricultural Policy (CAP) for tobacco. The premium
system is the mainstay of the tobacco CAP (Burr, 1992). A single
premium is set each year by the EU Council for each of the five
groups and three varieties. Premiums may be paid to registered
growers or grower groups. Their purpose is to make EU grown
tobacco more competitive on the internal market and to maintain
overall EU agricultural earnings. It is hoped that reducing premiums
will also slow the production of many unwanted types and varieties to
reduce over-production. However, it is believed in some countries,
e.g. Greece, that the EU premiums are not tied to quality and leave
little incentive to growers to put in the intensive labor that high quality
demands (Doolittle, 1995). Overall, prices for EU cured leaf are high
by world standards.
b
Payment
The need to pay the grower an acceptable, fair price governs his
continuance in tobacco growing. Paying the grower as soon as
possible after purchase of his leaf is equally important to sustain a
satisfactory relationship between him and the buyer. Based on this
review of marketing systems, it is considered that the industry largely
achieves both requirements. Payment is almost immediate in auction
systems and within 2 to 3 days in contracts with private companies,
leaf dealers and cooperatives. Some monopolies may take slightly
longer, notably government monopolies. In some instances, where
there is a very large number of small growers, part payment will be
paid within 15 days with the remainder within 45 days e.g. India
(Chandramouli, 1988).
Improvement of Marketing Systems
The marketing of cured leaf constitutes a significant cost, to both the
grower and the buyer. For the grower, it includes sorting, grading,
packaging and delivery to a place of pick-up or sale. In auction
systems, there is an additional fee for handling, classifying and selling
his tobacco. For the buyers, their cost includes personnel entailed in
the purchase of the tobacco, pick-up handling and transport from the
place of purchase to a central primary processing facility, data
handling and checking the integrity and weight of the leaf in the
package.
Where there are a very large number of small growers, e.g. India and
China, with large distances between farms coupled with poor roads,
collection and delivery costs are markedly increased, mainly for the
buyer. Again, there are significantly greater costs for the buyer under
marketing systems where there are a large number of auction markets
(and sale floors per market), e.g. USA (flue-cured, 53) compared with
Zimbabwe (flue-cured, 1), and Ontario, Canada (flue-cured, 2).
Zimbabwe's four auction floors, under one roof, provide an extremely
efficient, fully mechanized sales system. Growers' bales are checked
for the agreed delivery date and weight and the information is
computerized. The bales are off-loaded by mechanical conveyors
from the truck/lorry to minimize manhandling and pass across a
digital electronic scale, the data are entered on a sales ticket and
attached to the bales. The bales are then placed on 'railway' trolleys,
which can be mechanically moved in and out of the floors extremely
quickly to facilitate the total sale of 16 000 bales per day at a selling
rate of 6 seconds per bale. The grower finally receives a transcript of
every detail of each bale's sale. This is a service facility that provides
pleasant, well designed and restful amenities for both buyer and
grower, plus a modern cafeteria. While improvements can and will be
made, this is a system to emulate.
The marketing system in the USA has three components: the growers,
the warehouse or sales floor owners and the buyers. It has been clearly
stated that such a system, with a multitude of markets, inflates the cost
of marketing and hence the overall price of tobacco. With overall
production going down, considerable study and effort over many
years has, therefore, been expended to find means of improving the
situation and increasing the competitiveness of US tobacco in the
world market.

 Page 328
It is hoped that the main recommendations outlined by Marshall and
Donahue (1994) will be instituted over a period of time. These include
building well designed tobacco marketing centers with good lighting
and adequate space for tobacco display; to replace and reduce the
number of current markets; guaranteeing the entity or integrity of the
tobacco pile; complete reevaluating and revamping of the
classification system to meet buyer requirements; changing packaging
form, size and weight to improve efficiency in mechanized handling
for unloading and loading, pre-and post-sale; adopting a complete
electronic data management system, and so on.
Babcock and Johnson (1989) have stated that standards are used in
marketing systems to transmit information about products being sold.
Tobacco leaf buyers are saying that the current system for flue-cured
tobacco in the USA is not transmitting enough information about the
lot in today's technological world. High cost, variable tobacco leaf has
to meet virtually no objective specifications prior to purchase,
whereas the grower price of most crops in the world is related to
quantitative analysis of samples (Campbell, 1991). It is highly
desirable that such qualitative and quantitative standards closely relate
to those required in the final manufactured product.
In this regard, it has been shown that near infrared reflectance (NIR)
equipment can be used to determine moisture and leaf chemistry in
tobacco piles (McClure & Weeks, 1992). The possibility of its
practical application to measure these on the sale floor was envisioned
by Marshall and Donahue (1994). Such measurements could be tied in
with a subjective classification. This could be an aid to buyers
anywhere.
Improvements in marketing under contract systems with small
growers lie mainly with the producer. In most cases, their production
methods are specified by the buyer, e.g. varieties, fertilizers, crop
protection agents, the method of packaging and the materials used.
Where emphasis needs to be placed is on good sorting and grading by
the grower to meet buyers' demands and to reduce losses of cured leaf
in storage before sale.
It is quite clear that the whole tobacco industry throughout the world
is in a state of major change. As Cattelain (1990) has pointed out with
regard to the EU, producers must adapt to industry needs. Leaf
production and manufacturing techniques continually change to meet
consumers' demands or even government demands (limitation of
average 'tar' levels of cigarettes in the EU is 12 mg, effective January
1998). Leaf marketing is part of that change.
Price and quality/usability are intertwined. There is a very strong need
for full communication between growers and buyers everywhere to
ensure that both fully understand each others' problems and
requirements to move forward and achieve success.
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Anonymous (1991) Profile of the tobacco industry in Malaysia.
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Asociacion Rural de Interes Colectivo de Productores de Tabaco
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Australian Tobacco Marketing Advisory Committee (1995) Selling
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Babcock, B.A. & Johnson, P.R. (1989) Standards applicable to
Fluecured Leaf from Start of On-farm Preparation for Sale until Time
to 'Break the Sale.' NSCO Raleigh, NC.
Barton, H.C. (1995) Rebuilding leaf supply and demand in Central
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Folha Curado. Porto Alegra, Brazil, 1994. pp. 118.
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Campbell, J.S. (1993) Trends in tobacco leaf usability. CORESTA
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Campbell, J.S. (1994) Tobacco and the environment: the continuous
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Cattelain, P. (1990) E.C. Leaf producers must adapt to industry needs.
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Chandramouli, K. (1988) Auction system alters this Virginia market.
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Doolittle, D.E. (1995) Crisis looms: currency rates, quality questions
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Kenya Gazette, Supplement 45, Legal Notice No 235.
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John, G.A. (1994) Eastern Europe: Bulgaria. Tob. Int., 15 February,
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Malawi Tobacco Control Commission (1994) Grading and Marketing
of Tobacco in Malawi (all types). Malawi Tobacco Control
Commission, Lilongwe, Malawi.
Menzie, E.L. & Marshall, J.P. (1981) The Canadian and US tobacco
production, pricing, and marketing systems. MB 292. VPI and State
University, Virginia, USA.
Nobleza-Piccardo, S.A. (1995) Distribucion de Grados Tabaco.
Virginia Flue-cured (VFC) in Argentina. Buenos Aires.
Philippines National Tobacco Administration (1994) Standard Grades
of Philippine Flue-cured Virginia Tobacco. Also Memorandum
Circular (1995) Floor Prices. Republic of Philippines, (NTA), Quezon
City, Philippines.
Ridgeway, L. (1992) Rothmans: promoting quality flue-cured in new
environments. World Tobacco 130, 7780.
Tobacco Marketing Board (1994a) Classification of Flue-cured
Tobacco / Burley Tobacco. Harare, Zimbabwe.
Tobacco Marketing Board (1994b) Rules and Regulations governing
the Marketing of Flue-cured Tobacco/Burley Tobacco. Harare,
Zimbabwe.
USDA, AMS, TD (1986) Official Standard Grades for Virginia Fire-
Cured Tobacco (Type 21). Washington, DC.
USDA, AMS, TD (1989) Official Standard Grades for Flue-Cured
Tobacco (US Types 11, 12, 13, 14 and Foreign Type 92). Washington,
DC.
USDA, AMS, TD (1990) Official Standard Grades, Burley Tobacco
(US Type 31 and Foreign Type 93). Approved for reprinting in 1996.
Washington, DC.
USDA, AMS (1994) Operation Policies and Procedures relative to
the Marketing of Flue-cured Tobacco/Burley Tobacco. Washington,
DC.
Weeks, W.W. (1992) The importance of stalk position in harvesting,
curing, grading, and marketing of flue-cured, and the use of such
tobacco in cigarette manufacturing. Department of Crop Science,
NCSU, Raleigh, NC.

 Page 330

10B
Green Leaf Threshing and Redrying Tobacco
Bill Ward
Export Leaf Tobacco Company
Wilson, North Carolina, USA
Introduction
The first process step performed on tobacco after being purchased to
prepare for cigarette manufacturing is that of removing the stem. A
tobacco leaf contains a central stem known as the mid-rib which
supports and supplies nutrients to the leaf and is attached to the
primary stalk. Small stems, referred to as veins, radiate from the mid-
rib, providing structural support for the leaf tissue called lamina. Once
these stems are removed from the leaf, the resulting leaf product is
called strip. Stems, due to their size structure and chemical makeup,
have to be removed from the leaf and handled separately in the
manufacturing of cigarettes.
In the 1910's and 1920's, the stem was removed from the lamina by
hand. Production of lamina was 4.5 to 5.0 kg/hour per hand stripper. It
was not unusual to have more than 1000 people performing this task
day after day. In those days, leaf was marketed in bundles. Prior to
stripping, these were placed in a large conditioning room with
temperatures of 26 to 32°C and relative humidities of 75 to 80%. In
some locations, this conditioning was accomplished by using an
'ordering box'. This box contained a slow moving mesh apron and
steam nozzles. This process conditioned the leaf to a point where it
could easily be hand stripped from the stem without producing
considerable amounts of scrap. The stem and scrap were discarded as
waste byproducts and were not used in the production of cigarettes
until the late 1940s.
R.W. Coffee invented the first stemming machine in operation which
replaced hand stripping of tobacco in the 1930s and was the first step
in the mechanization of the stemmery (Richmond Dispatch, 1897). In
order to properly clean the stems and the scrap from the stemming
machines, the introduction of additional machinery was required such
as threshers, air legs, pneumatic separators and dust collectors. This
auxiliary equipment must be recognized as the forerunner of our
current threshing equipment. In the early 1940s, experiments were
conducted using this equipment to thresh whole leaf. The results of
these trials were so successful, green leaf threshing lines were
installed in drying plants throughout the leaf growing areas in the
United States. At this stage, a typical processing line consisted of an
apron type ordering machine, tie leaf breaker, ordering cylinder,
specially designed threshers for each position, air legs, pneumatic
separators, dust bags, convertible bundle drying machines and water
operated presses for packing tobacco into hogsheads. The capacity of
each threshing line was approximately 1000 kg per hour (Farmer,
1984).
In the early 1950s, the multi-thresher, 76.2 cm (30 inches) wide, and
the vertical classifier were introduced to the industry. The multi-
thresher was flexible in that it could be used in any position simply by
changing the teeth spacing and basket configuration. The classifier
was designed to incorporate closed circuit pneumatic conveying of the
threshed/partially threshed leaf from one separating unit to the other,
as well as recirculation of the lamina separating air. These features
limited the moisture loss from lamina and stem as they proceeded
down the processing line. While there were significant advantages to
this process, production rates were still only 1400 kg/hour.
During the past four decades, equipment has become larger and
somewhat more sophisticated while still employing the same process
principles introduced in the 1940s. Threshing lines today range in size
from 1.22 m to 3.66 m wide with multiple threshers in each position.
Production rates of the 1990s are now in excess of 14 000 kg/hour, a
10-fold increase in productivity since the early 1950s.
The basic concept of threshing has not changed considerably since the
1950s. The green leaf threshing (GLT) process during the 1970s to
mid-1980s was designed more for production than particle size.
Beginning in the mid-1980s, 'soft threshing' became the buzz word.
This concept is to apply multiple single rotor threshers (Fig. 10.1) in
the first and second positions, significantly reduce load rates (kg/cm
per hour) to each thresher, reduce rotor speeds and larger sized
thresher baskets. Threshing efficiencies are usually lower and this
results in lower degradation of

 Page 331

Fig. 10.1
Single rotor threshers.
the tobacco. This method of threshing, using more threshers of single
rotor design, has been given the name 'soft threshing' (Hotchkiss &
Ward, 1987). A typical processing line of the 1990s is illustrated in
Fig. 10.2.
Threshing Stages
A typical threshing line consists of four to five threshing stages. Prior
to the tobacco leaves entering the threshing process, they are manually
placed on a blending belt or into a bulk feeder (Fig. 10.3). The speed
at which this bulk feeder supplies tobacco to the process is controlled
by weigh belts to the desired production rate. Tobacco, once entering
the bulk feeder, is transported by conveyor to an ordering cylinder
(Fig. 10.4) where heat and moisture are applied. This is the initial step
in preparing tobacco to be threshed. A combination of heat, steam and
water is necessary to assist in opening the tobacco so all leaves have
an opportunity to be properly conditioned to the same temperature and
moisture content. Various types of tobaccos require somewhat
different temperatures and moisture regimes. Typically, temperatures
range from 45°C to 60°C, and moisture content from 18 to 21%.
Once the conditioned tobacco leaves the ordering cylinder it is then
transported by conveyor to either a screening cylinder or a vibrating
shaker (Fig. 10.5), primarily for the removal of sand; however other
materials such as stem and lamina fines are also removed. The
tobacco, still in whole leaf form, is then transported to proportioning
units which uniformly deliver equal amounts of leaf to searching
conveyors. These slow moving conveyors present the leaves Of
tobacco to individuals or, in some cases, automated searchers for the
purpose of removing off-color leaves, so as to meet customer quality
grades, and foreign material left in the product by the sand reel or
shaker. The tobacco continues to the final ordering cylinder where
combinations of heat, water and steam are once again applied to
optimize the threshing characteristics of the leaf being processed. The
tobacco continues to flow to a feed regulator and, in most instances,
where there are multiple threshers to a proportioning feeder and then
to the first position threshers. This is the beginning of the actual
process of removing the stems from the tobacco leaves and generally
is referred to as 'C & C' (cleaning and classifying), i.e. cleaning the
leaf from the stems and classifying (separating) the two products.
The tobacco is pneumatically transported beginning at the first
position thresher through the entire four, and sometimes five, stage
threshing process. The rotating teeth in the thresher strip the lamina
away from the stem as it passes through. Threshing efficiency in the
first position is about 65 to 70%, which means 30 to 35% of the
product is being introduced to the first position separators (Fig. 10.6)
requiring additional threshing in later stages. The product is conveyed
through the separators in series until it is either removed as lamina or
it enters the next stage threshers. Separation is achieved by vertically
directed air passing through the threshed product. A separator must
have sufficient separating horizontal area in order


 Page 332

Fig. 10.2
A typical processing line.

Fig. 10.3
Bulk feeder and dumper.
for the winnower to introduce the material in such a manner that the
stem free lamina does not become entangled with unthreshed or
partially threshed leaf and stem. The separating air must be vertical
and distributed uniformly across the entire area of the separator and
have sufficient velocity to lift and remove all stem free lamina from
the separator. The velocity (cubic feet per minute) of air to the
separator is dictated by the horizontal separating area and the air
velocity necessary to lift and remove accepted product.
Lamina, being lighter than stem, is lifted and extracted from the
process. The stem and lamina with stem still attached remain in the
process for additional threshing and separation. This same process is
repeated in each of the remaining three or four positions. Since the
GLT process is volumetric, approximate throughput rates for optimum
particle size results at a given stem content can be calculated by
comparing bulk densities of various tobacco types and to some extent
within type (flue-cured, burley, etc.). The leaves from the middle of
the tobacco plant usually provide excellent threshing quality. It should
be remembered the factors which affect the amount of stem left in
strip and the size of strip are thresher loading (throughput rate),
moisture content of the leaf being processed, number of positions in
the process, air velocity in the separators and the selection of basket
types in the threshers and the rpm of the thresher rotor.
The lamina removed from the process passes over a series of sieves of
various sizes to remove fine particles of tobacco and sand. These
'fines' usually pass another final screening operation to be properly
sized and to remove any additional sand (Fig. 10.5). A typical sieve
size would be 0.32 cm. The fines product remaining above the 0.32
cm sieve is usually reintroduced to the lamina product. The material
below the 0.32 cm sieve remaining above a secondary screen of
specified size is packed separately as a fines product and is used in
various ways. The material passing through the spe-

 Page 333

Fig. 10.4
Screening or vibrating shaker.
cified secondary screen is usually classified as dust/ sand and is
discarded.
Quality Control
During the threshing process, quality control is continuously sampling
the finished product for stem content remaining in lamina, as well as
its particle size distribution over various size sieves. Both stem
content and the particle size of the threshed lamina are important.
Data generated by quality control regarding both are continuously fed
back to the process operations manager via computer which trends
variable data for optimum process control. The results can change
depending on customer requirements. Particle size distribution (PSD)
for the lamina ranges from 65 to 85% for the total lamina above the
12.5 mm (1/2 inch) sieve. Coefficient of variation for the above 12.5
mm sieves is usually less than 5%. Also, the percent of lamina below
the 6.3 mm (1/4 inch) sieve is viewed as being an important variable
since about 70% of the material with a size of less than 6.2 mm
becomes noncontributory if cut in cigarette manufacturing with
regards to fill value (White, 1990). Some explanation at this point is
required to better understand the definition of stem and PSD.
 Fig. 10.5
Conditioning cylinder.
Prior to June 1994, almost every processor defined both stem and PSD
somewhat differently. Even though the basic analytical equipment was
the same, operating methods of this equipment were different. In May
1988, the Quality Control Committee of the Tobacco Association of
the United States began efforts to standardize, for the first time,
methods/definitions used in GLT operations regarding these two
variables of tobacco. In October 1990, both methods were given
CORESTA approval. Subsequent to CORESTA approval, application
was made to have these approved by ISO. Approval by ISO was
obtained in June 1994. While few companies continue to use other
internal methods, if one wants to compare quality control results from
different suppliers regarding particle size and stem content of strip,
methods such as ISO 12194 (PSD) and ISO 12195 (stem) should be
requested. There has been general acceptance of these methods by the
international tobacco community.
The CORESTA/ISO method for PSD requires a specified lamina
sample weight to be processed over a vibrating four deck shaker (Fig.
10.5) which has four critically sized sieves (2.54 cm, 1.27 cm, 0.62
cm and 8 mesh sieve to ASTM E-11 specifications). As the sample
passes over these sieves the various particle sizes are separated. The
weight separated by each sieve is expressed as a percentage of the
total sample weight,


 Page 334

Fig. 10.6
Vertical lifting separator.
including the calculation for the material passing through the 8 mesh
sieve.
Stem content is determined by processing a specific size sample of
lamina, usually the same sample used for PSD analysis, through a
stem tester which consists of a sample supply conveyor, thresher,
pneumatic transport, and a separation tower. The lamina is
continuously recirculated through the thresher and separator where all
lamina is eventually removed from any stem by the thresher; then the
separating tower efficiently separates the two products. At the
conclusion of the test period, the stems removed by this process are
accumulated and processed separately through a RO-TAP fitted with a
2.38 mm × 31.85 mm slotted sieve, 2.80 mm sieve and a 1.70 mm
sieve for sizing. As a result of this sieve analysis, the stems remaining
on the 2.38 mm × 31.75 mm slotted sieve are weighed and expressed
as a percentage of the original lamina sample weight and are referred
to as 'objectionable stem'; the stems remaining on the 2.8 mm and
1.70 mm are combined with the 2.38 mm x 31.75mm and this total
weight is expressed as a percent of the total lamina sample weight and
is referred to as percent total stem. The vein which passes through the
1.70 mm sieve is referred to as fiber and generally is not considered in
any results as it lacks definition. As previously mentioned, the details
of the methods can be found in ISO documents ISO 12194 and ISO
12195. Both methods are also referred to as CORESTA methods as
they were first recognized and approved as a standard by this
international tobacco research group.
Threshing and Redrying
Lamina from the threshing process is, in some operations, transported
to blending bins. Two bins are required although some prefer three
bins to accommodate short grade runs. These are operated usually on
an hourly schedule for filling and supplying the redryer. Blending bins
provide processing advantages, being located between the threshing
operation and redrying. Since the redrying process is extremely
sensitive to process interruption, bins act as a buffer to greatly
minimize the impact such interruption would have on the variation of
packed moisture content as the feed to the dryer is not altered. Bins,
by design, blend tobaccos, so reducing moisture variation to the dryer.
Now that the removal of stem from the threshing operation is
complete and the lamina product has been blended, it is necessary to
redry the products to a moisture content high enough to enhance the
aging process, but low enough to prevent oxidation to the point where
the product would not be acceptable for manufacture. Generally,
redried moisture contents of less than 15% are acceptable; however,
most tobacco strip is packed between 12 and 13% with an SD of <0.5.
There are three distinct periods in the development of redrying
methods. The period between 1865 and 1890 depended on the slow
and natural process and the use of steam coils. Usually the dealer
would hang tobacco on sticks until it was void of moisture.


 Page 335
This process of drying and packing required 8 to 10 months for
completion. During the 1890s, some progress was made where
various arrangements of steam coils were used for drying and a
steaming system was used for reordering the leaf when ready for
packing. It was not until 1880 that J.H. Prichard of Petersburg, VA and
C.A. and J.Q:. Jackson patented the 'Prichard dryer', complete with
steam outlets, ventilators and air intake. Several redryers were
patented after this; however, it was not until 1894 that J.K. Proctor,
inventor and manufacturer of textile machines, was commissioned by
John Wright, a tobacconist, to apply his recirculatory system of drying
cloth to the problem of redrying tobacco, By 1895, Proctor's invention
not only used the principal of a recirculatory system, it also included a
method for getting tobacco in and out of the dryer. It consisted of a
steel rack on rollers with room for hanging five tiers of tobacco on
sticks. It took about 45 minutes to dry and from 12 to 24 hours in the
ordering chamber to become pliable enough to pack in hogsheads in
whole leaf form. These redryers had a capacity of 68000 kgs per day
(Tilley, 1948). This capacity seems quite small by today's standards,
with rates well in excess of 10000 kg per hour and a moisture content
coefficient of variation of <3.0% for packed strip.
Blending Bins
Today, strip, directly from the threshing line or from the blending
bins, is conveyed to a sweep feed which distributes the tobacco across
the width of a pin feeder
 Fig. 10.7
Redryer.
which in turn conveys the strip onto the redrying apron. A typical
redryer (Fig. 10.7) is approximately 40 m long and 4 m across. A 3 m
wide metal conveyor called an apron travels full length, conveying the
tobacco through three zones, i.e. drying, cooling and reordering.
Several variables influence the drying objectives of packing uniform
moisture content and minimizing physical degrading (curling of
lamina by excessive heat). The moisture and temperature profile of
the product being redried, production rate at which the product passes
through the dryer (bed depth), tobacco density and type are all
important in producing a uniform product by this process.
A recommended bed depth maintained uniformly distributed across
the apron is approximately 10.1 to 12.7 cm. It is important to
remember ebbs and flows of tobacco being supplied to the dryer will
have a negative impact on drying performance as the bed depth will
vary down the length of the dryer.
The cooling system sections of the dryer are critical. Usually the
moisture content of the tobacco in these sections is 2.5 to 3.0% less
than the desired packing moisture. The product is cooled to 32 to
43°C to promote condensation of the steam on the much cooler
tobacco to be reabsorbed.
Now that the lamina product has been redried to customer
specifications, usually between 12.0 and 13.0% moisture content, it
leaves the redryer and is conveyed to the packing station. The packing
station is comprised of a hydraulic press, charger, scales, autostrapper
for cardboard cases and production scales. These are designed so as to
be able to press lamina into virtually any size or shape container (i.e.,
hogsheads, bales or cases).
Packing
There are basically types presses two of (one typical type is shown in
Fig. 10.8). The container is placed directly under the press and
charger. The charger, approximately 7.5 m tall, sits directly over the
package being filled. The package and charger are filled to the desired
weight. The tobacco flow is switched to a second charger so the
tobacco may be compressed into the package by lowering the
presshead. The second type of press works in a similar fashion, except
the package is located at a right angle to the charger. In this instance,
the tobacco is pressed from above as before, but is extruded into the
package by a horizontal hydraulic press. After the tobacco has been
pressed into the package, quality control takes a sample by

 Page 336

Fig. 0.8
Tobacco press.
inserting an auger, 2.54 cm by 55.89 cm long, assisted by an air
cylinder to retrieve a sample for various laboratory analyses such as
moisture content, nicotine, total and or reducing sugars, nitrates,
chlorides or any other analysis required by the customer. The size of
the auger is dictated by the number of analyses required. The product
temperature is checked at this point to ensure the product has cooled
enough to be stored without fear of oxidation. Once the sampling is
completed (15 seconds), the package is closed and proceeds to the
production scales where it is weighed and labeled, identifying the
content as to grade, account, date produced and weight. A typical
packing density for tobacco is 304.8 kg/m3. Even at this density with
a moisture content of 13%, 78% of the package is void of tobacco.
This is of some importance since tobacco, as other stored agricultural
commodities, on occasion requires some type of pest control measures
such as fumigation with an approved material such as aluminum and
magnesium phosphide (phosphine (PH3) or hydrogen phosphide is
evolved following exposure to atmospheric moisture) or some type of
modified atmosphere.
Stems from the threshing line go directly to a feeder which evenly
distributes them in a uniform bed depth across the redryer apron.
Drying time is approximately 9 to 12 minutes. Current apron stem
dryers, considerably smaller than lamina dryers, only have drying and
cooling sections as it is not deemed necessary to reorder this product
prior to packing since little or no breakage occurs at packed moisture
contents in excess of 10.0%. Temperatures in the drying and cooling
sections are automatically controlled with temperature sensitive
elements usually positioned on the air delivery side of the bed of stem
(Farmer, 1984).
Once the stems have been dried to the desired moisture content, they
are picked up pneumatically and transported .through a corrugated S-
shaped section of duct. Any small pieces of attached lamina which
were not removed in the threshing process are easily removed as the
small pieces of lamina are now quite brittle from the stem drying
process. They are separated from the stem with an air leg and
transported to shakers for proper sizing before being packed as fines.
The stem passes over a vibrating conveyor which can be fitted with
various size perforated plates for grading stem by size. Small pieces of
stem sieved out of the process are used as feedstock for reconstituted
tobacco products. Once stems have been sorted by size, they are
packed in any number of package types such as hogsheads, tersa bales
or corrugated cases and are ready for shipment to the customers,
storage or a reconstitution facility.
The fines material is continuously being pneumatically conveyed from
the threshing operation and can be dried on the way to the sizing
sieves by the introduction of heat in the transporting duct work after
sizing. In some instances, fines are sized and reintroduced to the strip
prior to the strip dryer. Fines material, being the thin and friable
portions of the leaf being threshed, are easily dried in this manner.
This product can vary considerably in size depending on the size of
the sieves being used for screening the strip product. The material
passing through the second screen is routinely used as feedstock for
reconstituted tobacco products.
Summary
The process of threshing/separation, blending, redrying and packing is
now complete. Products are sent to storages where they are aged for
several months to enhance the smoke characteristics and are
ultimately shipped to manufacturing.

 Page 337
References
Farmer, W.B. (1984) Leaf processing from conditioning to prizing.
Tob. Int., February, 3.
Hotchkiss, C.A. & Ward, W. (1987) Soft threshing, the evolution
continued. Tob. Reptr., 114(5), 246.
Richmond Dispatch, 1 January 1897.
Tilley, N.M. (1948) Bright Tobacco Industry 18601929. University of
North Carolina Press, Chapel Hill, NC.
White, M. (1990) Translation of uncut to cut particle size. Tob. Reptr.,
17(7), 1820.

 Page 338

10C
Tobacco Storage
L. Ryan
Philip Morris Europe
Neuchâtel, Switzerland
Introduction
The 8 million tons of tobacco grown globally are stored in warehouses
around the world for varying duration. Tobacco will be stored in
several locations as it passes from the grower to the finished product
manufacturer. Unfortunately, as will be seen by the few references,
this topic is usually given only cursory treatment in the limited
number of books devoted to tobacco. This is surprising since the value
of tobacco in storage, globally, at any given time is in the order of
$US 12 billion (109).
Tobacco types are dealt with elsewhere in this monograph (Chapter
5), but for simplicity we will consider fermented tobacco and aged
tobacco.
Cigar tobacco and dark air-cured tobacco are fermented, as is the
semi-Oriental tobacco. This is true fermentation as opposed to the
natural 'fermentation' of Oriental tobacco, a process which is more
akin to aging (Akehurst, 1968). Aging is a slow change once the leaf
is packed and in storage, thus all tobacco may be considered to age,
but the term is generally used only for cigarette tobacco such as flue-
cured, Oriental, Maryland and burley. Aging may be considered a
mild state of fermentation (Tso, 1990). Processed tobacco is aged,
usually for 1 to 3 years, to improve taste, aroma and quality
characteristics. The duration of aging is governed by logistic
constraints (see section, this chapter, Handling and Control). Thus,
aging occurs during storage where tobacco with a moisture content of
10 to 13% will lose 1 to 2 percent of dry matter for flue-cured and 3 to
4 percent for dark air-cured.
Aging results in changes of aroma and flavor characteristics. These
changes are thought to be associated with starch conversion to sugars,
sugar and amino acid reactions, reduction in amino acids, increase in
water soluble acids, increase in pH and loss of nicotine. With more
uniform packed moisture content, aging today is different to the pre-
redryer days. If tobacco is packed too wet or with wet spots the aging
may become a true exothermic fermentation, with the resultant heat
causing either caramelization of the tobacco (blackening) or even fires
from spontaneous combustion. Therefore, warehouse managers are
reliant on their suppliers to have the redrying process under control
and supply tobacco at consistent and uniform packed moisture.
The term 'sweats' is better forgotten; it describes the fermentation
process and was used to describe aging, thus, only serving to confuse.
Tobacco is stored in packages such as hogsheads, cases, or bales (Figs
10.9, 10.10 and 10.11). Hogsheads are wooden barrel-type containers
with approximate dimensions of diameter 120 cm and height 140 cm
and they contain between 385 and 455 kg tobacco. Cardboard cases
will contain about 200 kg; burlap bales, usually for Oriental, are
packed to hold between 30 and 50 kg. Flue-cured and burley bales
used by some manufacturers in lieu of hogsheads contain between 400
and 460 kg. Thus, warehouses must be equipped to handle all types of
packaging in an economic manner.
 Fig. 10.9
Cases or cartons of tobacco in storage.
The following are some guidelines for storing tobacco to fill a void in
the literature and assist those who wish to improve storage conditions
or build a new warehouse to optimal standards. The sections deal with

 Page 339

Fig. 10.10
Hogsheads of tobacco in storage.

Fig. 10.11
Bales of tobacco in storage.
types of warehouses, warehouse construction, warehouse
environment, tobacco handling and control and preventive quality
assurance. The more commonly found tobacco is discussed in this
section and not storage of tobacco such as perique, which is pressure
packed for 2 years.
Types of Warehousing
Originally, the main problem confronting warehouse managers was to
control the tobacco 'sweats'. When tobacco is packed at too high a
moisture content and then subjected to a high ambient temperature,
the result is mold. Thus, early warehouse designs concentrated on
maintaining even temperatures by ventilating 'open air' warehouses.
These open air warehouses often had basements, flat roofs and
louvered sides and became known as 'Dutch Barns', an adaptation of
the Snow curing barn design (Tilley, 1948).
With improved uniformity of moisture content at packing due to
improved redrying technology, warehouses became less open. Indeed,
the ability to close and seal warehouses became an important design
feature with the advent of phosphine fumigation in the early 1970s
(Ryan, 1995). Warehouses are now simpler in design and may be
divided into on-farm, auction, processor, dealer, manufacturer and in-
transit.
On-farm storage is ideally used only during the curing phase,
fermentation normally being handled by processors. In reality, the
tobacco may stay on-farm for days or weeks until it enters the
marketing system. Unfortunately, there may be considerable quantities
of a farmer's crop that remain on-farm from one season to the next or
longer. This 'carryover' tobacco may appear in the marketing system at
the beginning of the next season.
Tobacco stored on-farm should be retained in the newly sterilized
curing barn for flue-cured (Akehurst, 1981; Collins & Hawks, 1993)
and otherwise, in a clean storage shed where tobacco has been isolated
from such materials as agrochemicals and fuel. Sanitation is
important: this shed must be thoroughly cleaned and screened to
prevent insect, bird and rodent entry. Wherever possible, avoid
problems with carryover of tobacco by at least processing the tobacco
to usual storage moisture. Lengthy on-farm storage may lead to off
odors, unsound condition and mold. Studies with burley stored as
farmer bales or bundles at 22 to 30% moisture for the summer months
showed off odor was common (Bunn & Henson, 1978; Walton, et al.,
1985).
Auction warehouses may store the tobacco for days before the day of
sales. These warehouses must have a strict sanitation standard in place
and must follow the basic storage instructions given below. The
occasional practice of storing farmer tobacco in these warehouses
should be avoided.
Processors will normally store the nonredried or farmer or 'green'
tobacco for weeks prior to manipu-

 Page 340
lation or redrying. The rule here will be for standard warehouse
practice and that this green tobacco is physically separate from the
processed tobacco. This is mainly to avoid cross-infestation by
insects: the risk of receiving infested tobacco from farmers' carry-over
is ever present. A few processors keep green tobacco in cold storage
to allow evening of the supply to the stemmery and, thus, year-round
use of the factory. Green tobacco, otherwise, is stored under the same
conditions as redried tobacco (see the section on Warehouse
environment). Because the moisture content is less uniform, a
monitoring program will ensure damage and loss are avoided.
Dealer warehousing is used here to refer to all tobacco stocks that
have been processed but not yet sold or distributed to the
manufacturer. This includes processors who hold the tobacco in
storage, government stabilization or 'pool' stocks, government reserve
stocks and the stocks held by leaf merchant companies. In all cases,
the warehouse requirements are as for any long-term storage.
Manufacturers of finished tobacco products require warehouses that
meet all requirements for long-term storage.
In-transit storage of tobacco is usually avoided as it is unnecessary in
most cases. Usually, the tobacco is packed into containers, trucks or
the like, for shipment direct to the customer. In-transit storage may
occur as tobacco transits a custom bonded warehouse. This transit
may be swift or require days if problems appear in shipping
documentation. Such warehouses are often at a port and usually are
not designed for tobacco storage. Ports are often infested With insects
and rodents (Ryan, 1995) and, as such, storage here should be
avoided. Even the in-transit storage of containerized tobacco at ports
should be avoided.
Tobacco may be 'stored' in land-sea containers as it moves from
supplier to customer for several days to even several weeks. Thus,
such containers must be leak proof, odor-free and screened against
insect and rodent entry. Care is needed when screening containers
where special ventilation is required. The same is true for tobacco
'storage' in-transit in trucks or ship holds.
Warehouse Construction
The following gives guidelines for ideal construction. Situate tobacco
warehouses away from industrial and residential areas with a
minimum of 20 m to the fence line. Ensure there is no risk from:
floods away from flood zones with good drainage
explosion away from fuel or gas storage
odors away from chemical factories, abattoirs, etc.
General construction requirements such as the type of electrical
supply, water and sewage system, fire prevention walls and devices,
floor loading for forklifts, roofing, etc., must conform to the local (or
national) authority requirements and often those of the insurers. Such
considerations are outside the scope of this document. However,
below are some common-sense and general requirements that the
personnel building the warehouse must consider. Warehouses may be
of several floors but a single storey is preferred to simplify tobacco
movement without elevators or conveyors. Equally, basements are not
required and where they already exist are left empty; tobacco is not
stored in basements due to the difficulty with moisture control.
a
Landscaping and Grounds
Gravel or concrete a perimeter around the warehouse greater than 1 m
to reduce insect risk and facilitate sanitation inspections and security
patrols.
Plant shrubs and trees, if desired, greater then 5 m from the
warehouse; preferably, plant only lawn and flower beds to reduce the
risk of birds and insects.
Place trash collection area greater than 10 m from the warehouse in a
walled area with a drained concrete base.
Aim sodium vapor type exterior lights at work areas from several
meters away. Sodium lights attract less insects and placement means
insects are not drawn to work areas.
Position mercury vapor type perimeter lights greater than 20 m from
the warehouse. Use sodium if possible.
Use low watt, yellow lights for other exterior lighting such as
doorways.
Ensure good drainage to avoid any standing water and ensure drains
are accessible for cleaning or rodding.
Maintain a greater than 2 m border outside the perimeter fence cleared
of vegetation or as a lawn to facilitate inspections.
b
Specifications for Construction
Finish all construction and installation work to avoid unfinished holes
which may provide rodent entry points.

 Page 341
Provide adequate lighting: use the standard used in factories to
facilitate sanitation activities and inspections.
Install no false ceilings or false walls: such voids are difficult to clean
and provide harborage for pests.
Avoid underground ducting; ensure drains are accessible, flushable,
and roddable.
Provide physically separate areas for green and redried tobacco to
prevent cross-infestation.
Screen all openings such as windows with 8-strand/cm mesh screen;
install air curtains or flexible strips over doors to prevent insect, bird
and other animals' entry.
Install removable mesh covers on all drains, inside and outside, to
facilitate cleaning and prevent possible pest exit from the drains.
Install exit doors of good fit, self closing, with detecting sensors if
propped open to prevent pest entry and for security.
Install roll doors with a flexible bottom 'seal' and T extensions to fit
rail tracks to exclude rodents.
Design out ledges of any kind (this , not, ¬, or ^) inside or outside
the warehouse to avoid additional surfaces to clean and limit potential
roosting sites for birds.
Ensure all walls are smooth inside and outside, no rough brickwork,
etc., to reduce dust accumulation.
Require adequate damp proofing under the main slab and between the
slab and walls to avoid moisture ingress.
Construct to obtain maximum water tightness, to avoid leaks from
walls or roof and avoid condensation.
Construct to obtain maximum gas tightness to facilitate fumigation (if
necessary), paying close attention to wall/floor and wall/roof
junctions.
Seal expansion joints flush with the floor to reduce dust
accumulations and facilitate fumigation.
Use concrete for the floor which does not generate cement dust.
Paint exterior white or light colored to facilitate temperature control
(Joost & Beard, 1985).
Allow no food on-site during construction and place the construction
crew canteen at the perimeter to reduce peat attraction.
Drain lift shafts to allow wet cleaning.
Use metalwork shaped like Ç, O, L or , not È V, ^ or , to avoid
dust build-up.
Minimize the amount of direct sunlight from windows or avoid the
use of windows entirely to facilitate temperature control.
Appropriate safety procedures and equipment such as fire
extinguishers must be provided.
c
Tobacco Inspection Area
This area allows visual inspection of the incoming tobacco and should
be a separate, well defined area.
Separate at least 100 m2 of dry, clean, covered (roof) space.
Paint walls a light matt color to avoid reflections.
Avoid direct sunlight but situate to utilize natural northern sunlight.
Provide an office area, washroom and toilet facilities in or close to the
area for inspection personnel.
d
Service Areas
Separate an aerated area for forklift storage and forklift battery
recharge.
Separate the kitchen and restrooms from the warehousing area by two
sets of doors for hygiene and to limit pest movements.
Completely tile restrooms and eating areas to facilitate cleaning.
Provide a separate area to store cleaning equipment.
e
Fumigation Areas
Fumigation is discussed below; however, when constructing
warehouses a number of points should be considered. Fumigations are
conducted in available containers such as land/sea or railcars, under
tarpaulin sheets, in specially built rooms or chambers, or indeed, the
whole warehouse may be fumigated. Depending on the local logistics
one or more or none of these options will require the following
actions.
Containers
Provide a secured area for such fumigations. The area is sized to
accommodate the number of containers foreseen. Security will require
a cordon at least 20 m around the site. Obey local authority
requirements.
Concrete or pave the area slab and provide good drainage.
Cover the area with a roof to allow aeration during inclement weather.
Tarpaulins or Sheets
Stack tobacco in standard stack volumes to allow standard sized
sheets and fumigant dosage.


 Page 342
Allow at least 1 m between stacks and walls to allow fumigators to
place the sheets properly and seal to the floor with sand or sand
snakes.
Plan for stacks fumigated under sheets to be cordoned to exclude
workers not involved in the fumigation. This usually means the
warehouse is closed during the sheet fumigation.
Ensure an even concrete floor, without drain covers, expansion joints
or other openings to prevent gas leaks.
Chambers or Fumitoriums
Such constructions require specialist knowledge, and an experienced
contractor should be involved. The following are only brief guidelines
for using phosphine (PH3) or hydrogen phosphide. Chambers should
only be considered if fumigations are to be conducted regularly.
Obey local or national rules governing the construction, specification
and pressure testing of such chambers.
Aim for maximum gas tightness with gas-tight walls and a well
sealing door, therefore, without windows.
Install a fan to aerate the chamber; the exit gas may be passed via a
filter to 'scrub' the air. Such scrubbers are usually activated, metal-
coated charcoals.
Install all electrical devices external to the chamber to avoid the
corrosive effects of phosphine.
Allow an airspace around the entire chamber.
Whole Warehouse
Stack fumigations are preferred since they are simpler, use less
fumigant and are, therefore, less expensive. The subject of whole
warehouse fumigation is dealt with in detail elsewhere (Ryan, 1995).
The basic requirements are:
Construct to allow simple closure and sealing of the warehouse to
minimize the amount of temporary sealing.
Ensure at least a 20 m security perimeter around the warehouse.
Respect all national and local laws regarding fumigation along with
specific instructions from the purchaser.
Train personnel to liaise with the fumigating contractor.
Warehouse Environment
a
Temperature and Humidity
During storage, environmental conditions are stabilized to avoid mold.
Mold will result in off odor and unusable tobacco which will have to
be destroyed. Mold is easily avoided if the temperature and relative
humidity (RH) are stabilized and low. Of course, it is essential that the
tobacco is packed at a uniform moisture level of 12 to 14%. Wet spots
or pockets of high moisture in a tobacco package will result in
molding the entire package. Tobacco that is 20 to 25% moisture will
invariably result in mold. Equally, the storage must have no leaks,
damp areas nor condensation (Samfield, 1980). A realistic standard is
ideally, average daily conditions of:
Temperature: 15 ± 5°C with a maximum of 23°C
RH: 65 ± 5%
Records are kept to assist with necessary corrective actions based on
data. Such records may be from electronic data loggers or the chart
recording thermo-hygrographs.
Maintaining these conditions is easy if the warehouse was constructed
with this in mind. Depending on the local weather conditions,
additional considerations may be necessary such as recirculation fans
or even cooled air fans. Based on the records, an engineer will be able
to adjust the building to meet these conditions.
In a number of countries tobacco is stored outside under sheets where
it is exposed to high summer and freezing winter temperatures. In
these cases, it is advisable to manufacture these tobaccos quickly.
Tobacco seeds must be stored under controlled temperature and RH
but, as seeds are more robust than leaf, they may even be freeze-dried
(Main & Gwynn, 1967).
b
Odors
Store only tobacco in a given warehouse and investigate what
commodities were stored previously. Persons with 'a good sense of
smell' should spend time in an empty warehouse before storing
tobacco to detect off odors. Personnel such as those with experience
as flavorists are ideal for this job. Off odors may well be detected if
odorous materials such as animal skins, rubber, cotton or chemicals
have previously been stored.
Forklift trucks or other vehicles that enter the

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warehouse must be powered by battery or an odorless fuel. Store other
materials such as pallets or used packaging materials away from the
tobacco.
Handling and Control
Inventory control is essential to ensure proper long-range planning
and optimal tobacco movement in relation to blend requirements in
production (Samfield, 1980). Blend components are usually 'split' as a
precaution in case of fire: larger lots are separated into different
sections (Tilley, 1948).
If one imagines company A produces 100 billion (109) cigarettes each
year with some 800 mg tobacco per cigarette, then 80 million kilos of
tobacco are needed each year. Assuming only a 2-year blend stock and
an average price of $US 2.00 per kilo, this represents $US320 million
in storage. This value in store provokes more than a passing interest
from the insurance company, who may themselves have certain
requirements regarding the disposition and protection of this asset.
a
Warehouse Routine
The storage and control of the tobacco stocks and the flow to
production sites are complex tasks made simpler with the advent of
computer programs (Camp, 1990). The following is a reasonable
scenario.
Tobacco arrives at the warehouse in containers, trucks, railcars or by
other means. If sealed, the seals will be checked as intact and recorded
as such. If monitored in-transit with pheromone-baited traps, the
information is recorded. Each tobacco package will have the
company's specific markings that identify the tobacco, total and type
of packages in the lot, type of tobacco, grade, crop year, origin
company and location, weight as gross and net, destination section of
the warehouse and other details which will be checked on arrival and
the date added. This information is sometimes as a barcode read
directly into the computer database. Inspection and quality assurance
(QA) data will be added to the database later (see following).
The container is off-loaded, by forklift or manually, and the packages
stacked in designated areas in uniform stacks. Forklifts with large side
clamps operated via adjustable pressure are preferred. It is important
that personnel involved in this unloading have basic tobacco training
on tobacco types, importance of odors, recognition of mold or damp
and such things as recording pheromone trap data and procedures to
follow if fumigant is found.
Tobacco is stacked on pallets and, depending on the package type may
be eight high for cartons (Fig. 10.9), with the upper three cartons
staggered, six high for hogsheads (Fig. 10.10), and eight high for flue-
cured and burley bales. Oriental bales are often stacked on racks. If
racks are not used the pallet configuration and stack height will
depend on the bale weight and whether placed flat, on the side
(plakka), or on edge during aeration (kilitch). A guideline for plakka
bales is: six high per pallet for bales less than 30 kg and four high per
pallet for bales more than 30 kg (Fig. 10.11). Stack height for Oriental
bales is usually up to three pallets high with supporting racks. 'On the
joggle' storage of hogsheads, that is, hogsheads on their side as
opposed to the 'normal' storage as a vertical cylinder, is with a stack
height up to four. This practice is decreasing due to the logistical
difficulties of extracting test hogsheads from joggle stacks. Green
tobacco may be stored as 'farmer bales' on a racking system or as
'extruded' bales: four high. Stacks are of uniform volume with an 80
cm wall perimeter, for inspections plus safety measures and an
aeration spacing of at least 10 cm every 6 m. The spacing will be 1 m
everywhere one suspects fumigation may be necessary (see above).
A portion of the tobacco is set aside for inspection by leaf specialists
to confirm the shipment meets contractual requirements. Such
inspection is in the specially prepared area (see above) and takes place
within days of arrival. Inspection is of a certain percentage of each
shipment. Inspection usually includes checks of visual quality,
soundness, aroma, weight, package integrity and whether recycled
packing or not, presence of odor, stains, or water damage and signs of
infestation by insects. Corrective actions will be company dependent,
but if a lot does not conform to the customer's requirements it may be
rejected. It may be possible to reject only certain packages, in the case
of water damage, for example. Usually, an investigation will proceed
to identify the problem cause and, therefore, a solution. In the case of
infestation, lot rejection or refumigation are options.
A portion is set aside for QA monitoring. The amount depends on
company requirements, but may be 1%. The type of testing may
include strip

 Page 344
size distribution, foreign matter quantification, stem content, moisture
content, tar and nicotine content and a battery of chemical analyses,
including pesticide residue testing.
Tobacco will remain in storage until required for manufacturing,
which may be only months or up to several years. Most companies
have an optimal crop mix where a 2-or 3-year overlap is planned.
However, given the vagaries of climate and, therefore, agricultural
crops, length of storage may vary.
Preventive Quality Assurance
a
Sanitation
Guidelines for constructing warehouses are given above; in many
cases the requirements are to obviate or at least alleviate the need for
cleaning. However, as tobacco packages are moved in and out of the
warehouses there is, unfortunately, some spillage and tobacco dust
production. If left unattended this offers a ready source of food for the
two main tobacco pests, the tobacco moth Ephestia elutella and the
cigarette beetle Lasioderma serricorne.
These insects have been discussed in Chapter 7B and the whole
subject of infestation, prevention and control is covered in a review
(Ryan, 1995). Given the importance of sanitation in tobacco storage a
little repetition follows.
The work of the cleaning staff must be facilitated. In particular, access
should be improved because areas difficult to clean will probably not
be cleaned properly, if at all. Examples of such areas are gaps between
storage pallets and walls, and the spaces underneath and behind
machinery and fixtures, such as below weigh scales. Store only
tobacco in a tobacco warehouse.
Where applicable, all 'auxiliary' machinery (e.g. fork-lift trucks,
chargers, lowerators, elevators, conveyors, air-conditioning units, etc.)
should be given extra attention during cleaning and removed when not
required.
Regular attention should be paid to drains, gullies and sinks for they
can be an important source of mold, flies, cockroaches, other insect
pests and rodents. They should be cleaned and inspected frequently,
and any defects found should be remedied without delay. Waste
materials and tobacco which accumulate in bins, etc., should be
removed daily. The bins should be cleaned regularly and no tobacco
stored on the production floors.
All cleaning personnel should be briefed as to the nature of tobacco
pests and encouraged to report their finding of anything suspicious.
The objective is to underline the importance of their work in
contributing to hygiene and pest control and to gain their help in the
early detection of infestations. Industrial vacuum cleaners, with
numerous suitable attachments, will be necessary to do an efficient job
of cleaning. Air blowing has no place in a sanitation program: this
exacerbates the situation by causing dust movement to inaccessible
areas. Suitable vacuums have a 3000 W motor capable of moving 7
m3 air per minute. Cleaning personnel will quickly identify
attachments needed to enable them to meet the cleaning specification.
Each warehouse must prepare a cleaning schedule which clearly
defines the procedures and roles of all personnel. A regular inspection
and auditing system should be carried out, documented and action
taken by identified personnel. How to develop a sanitation program is
described in detail elsewhere (Ryan, 1995).
Cramped storage should be avoided. There should be ample space for
the product stored with adequate access, light and ventilation. This is
normally required by law and fire insurance companies.
Stacks should not be built into walls or into corners. They should be
kept clear of doors, windows and ventilators. Stacking plans should be
marked out with painted lines on the floor, leaving at least 80 cm
around each tier for cleaning.
Corners and wall/floor junctions should be painted white to encourage
effective cleaning and facilitate inspections.
Good stock rotation is important, the rule here is first in, first out
(FIFO).
Cases and bales are stored on pallets or shelves. Since pallets
themselves are a source of infestation and cross-infestation, they
should be subject to regular inspection and cleaning: adhere to
industry standards for pallets.
Immediate repair of broken windows, damaged roofs and doors and
overall weatherproofing are necessary for good warehousing practice.
Doors, windows, fan vents, air bricks and hoist apertures may provide
pests with easy access to buildings unless they are soundly
constructed and properly maintained and cleaned. Pipes, drains, cables
and ducts passing through walls and floors must be completely sealed
to deny access of all pests.
If a window must be open, an 8 strand per cm screen will prevent
insect entry. If a door must be open, an air curtain will prevent insect
entry. An air curtain is a fan blowing air down the length of the
opening, directed outward at a 45° angle, at 40 km/h.

 Page 345
b
Monitoring
Infestation may begin post-curing. Carry-over or tobacco stored on-
farm poses a great risk of infestation. Rapid transfer from the farm to
processing is preferred. The farmer should store tobacco in clean
storage barns with screened windows and doors. A pheromone trap
should monitor for any potential infestation. Industry groups and
extension agents should stress the need for sanitation on farms. On-
farm storage, before marketing and during one or more years of carry-
over in infested barns, results in massive infestations entering the leaf
supply channels. Improved sanitation during on-farm storage and pre-
market storage is a long overdue must and may not be over-
emphasized.
Because of the possibility of bringing infested tobacco into the
warehouses and the risk of feral insects gaining entrance, a
comprehensive pheromone-baited trapping program should be in
place. Such monitoring is necessary when ambient temperatures are
above 15°C, when the insects are able to fly. More details are given in
Chapter 7B and elsewhere (Ryan, 1995).
c
Fumigation
Guidelines are given above regarding construction and the subject is
discussed in Chapter 7B. Fumigation is a specialized task and requires
qualified, licensed, experienced contractors or personnel. Companies
who do fumigate should appoint personnel to be trained to interact
knowledgeably with contractors. Tobacco fumigation is potentially
dangerous and requires extensive safety precautions (Ryan, 1995). A
few key points to remember are:
Use only packaged fumigants such as 'bags', 'ropes' or 'plates' to avoid
fumigant dust.
Opt for magnesium phosphide formulations in preference to
aluminum phosphide to allow a standardized 4-day fumigation period
at temperatures above 7°C. When using aluminum adjust the duration
according to temperature.
Monitor the concentration of phosphine during the fumigation and
ensure 200 ppm was achieved for more than 48 hours and that the
final concentration is above 100 ppm. The longer the fumigation
period the better.
Obey all safety rules and laws: if in doubt, don't fumigate.
Summary
This section has included a general outline of the facilities that are
needed to properly store tobaccos. The exact plans that may be
developed are left to the particular individuals and their situation. At
times, it may be necessary to convert an existing facility that was not
designed for this purpose or renovate one that has been used to store
tobaccos. Frequently, due to the age and condition of the warehouse
(storage sheds), they are in need of repair. This is a special problem
when the facility is to be fumigated. The stored tobacco is worth large
sums of money and it is logical that the facility in which such a
valuable commodity may be stored for years be a suitable one.
References
Akehurst, B.C. (1981) Tobacco 2nd edn. Longman, London.
Bunn, J.M. & Henson, W.H., Jr (1978) Environmental requirements
for storage of baled Burley Tobacco. Trans ASAE, 21, 96771.
Camp, R.E. (1990) Simulation determines storage requirements at
Reynolds Tobacco Company. Ind. Eng., 22, 2831.
Collins, W.K. & Hawks, S.N., Jr (1993) Principles of Flue-cured
Tobacco Production. North Carolina State University, Raleigh.
Joost, T.E. & Beard, T.J. (1985) Thermal stratification modeling and
performance in two agricultural warehouses. In: Solar Engineering.
pp. 97103. ASME, New York.
Main, C.E. & Gwynn, G.R. (1967) Seed storage in tobacco. Tob. Sci.,
11, 567.
Ryan, L. (1995) Post-harvest Tobacco Infestation Control. Chapman
and Hall, London.
Samfield, M. (1980) Research and Manufacturing in the U.S.
Cigarette Industry. Lockwood, New York.
Tilley, N.M. (1948) Bright Tobacco Industry 18601929. University of
North Carolina Press, Chapel Hill.
Tso, T.C. (1990) Production, Physiology, and Biochemistry of
Tobacco Plant. Ideals Inc., Beltsville.
Walton, L.R., Casada, M.E., Taraba, J.H., et al. (1985) Storage of
burley tobacco in bales and bundles. Trans ASAE, 28, 13011304.

 Page 346

Chapter 11
Cigarette Manufacture
11A
Tobacco Blending
Phil Fisher
Tobacco Consultant
Louisville, Kentucky, USA
Introduction
A successful tobacco blend is a product that provides consistent
consumer satisfaction at a fair competitive price. Tobacco blending as
it was practised from the 1950s to the late 1970s was discussed by
Akehurst (1981). Brooks (1953) described products made by the
North American Indians in which the sharpness and taste of Nicotiana
rustica leaves were reduced by adding other products. Salzsman
(1963) described tobacco blending as being like a symphony orchestra
where the total sound overshadows the sounds of individual
instruments. Cigarettes that provide the consumer with what they
want, vary considerably around the world. Straight Virginia blend
cigarettes are preferred by some people, while others prefer straight
Oriental, fermented air-cured products containing various additives or
many combinations of the above. The most preferred product,
universally, is the American blended product. This section will deal
with the blending of the American style product.
The content of a typical USA blended product is shown in Table 11.1.
Blending actually begins with the geneticist who selects tobacco from
various germplasm.
Table 11.1 The constituents of
a typical American blended
product.
Tobacco Content (%)
Flue-cured 2535
Burley 2535
Oriental 315
Cut-rolled stem 310
Reconstituted 1025
Total 100%

This important process insures uniformity of the product, grown with

the desired characteristics. In the US alone, there are approximately
40 to 50 accepted flue-cured varieties and 35 to 45 accepted burley
varieties. These varieties have expanded around the world and,
combined with germplasm from other local countries, give the blender
a variety of tobaccos from which to choose. Those selections,
combined with other tobacco types such as Oriental, fermented air-
cured and air-cured, are the basic ingredients for a tobacco blend.
All tobaccos can be classified into three basic groups:
(1) Flavor grades these impart a specific flavor contribution to the
smoke with the desired degree of irritation.
(2) Modifier grades these, on their own, are not positive flavor
contributions, but, when combined with other types, e.g. Oriental,
impart a pleasant taste and aroma with the desired degree of irritation.
(3) Filler grades these help fill up the cigarette tube without having
an adverse effect on taste or chemistry, thus they increase the filling
power (it takes less tobacco to fill the cigarette tube).
All tobacco types will fit into these three categories. Each individual
tobacco company has a range of grades that fit into the above three
categories. The criteria are the degree of flavor contribution, stalk
position, ripeness, price and country of origin.
Flue-Cured
Flavor contribution has a direct relationship to stalk position. The
flue-cured plant can be divided into four categories, beginning at the
bottom:

 Page 347
(1) sand lugs, primings
(2) cutters
(3) leaf
(4) tips
The sand lugs/primings offer the least flavor contribution. They are
lowest in nicotine content, but, generally, have the highest filling
value. Filling value is defined as the amount of tobacco occupying a
given space. The most important characteristic of primings is that they
do not contribute any off taste characteristics. Because of their lower
nicotine content, the blender can use good filler to maintain final
blend nicotine content, yet not contribute to off taste. The price of
good filler tobacco is generally low, so the blender can use this
material to lower the final blend cost. Good filler can be produced in a
wide range of countries and under various climatic conditions.
Filler tobacco also expands much better than other stalk positions,
thereby further lowering blend cost. There are several methods for
tobacco expansion. Once again, expanding tobacco is a method of
increasing filling power.
The cutters are generally used as modifiers or in a combination of
modifier/filling grades. Again, off taste is a major criterion to avoid.
Cutters generally absorb casing and flavor material effectively. They
are usually in the nicotine range of the finished product, but
sometimes can be higher. Cutters have the unique characteristics of
blending very well with other tobacco components. Each type of
cutter has its own special characteristic. They are generally not as
shatterable as primings/sandlugs, thereby giving a higher yield of strip
product. A good cutter can even impart some flavor contribution,
depending on the cultural practice and degree of ripeness.
The leaf stalk position is the biggest flavor contributor. Tobacco from
this stalk position is the basis for the 'American blended cigarette
taste'. The nicotine contribution is approximately 0.50 to 0.65%
higher than the cutters, 1.00% higher than the lugs and 0.20 to 0.50%
lower than the tips in flue-cured types. The degree of ripeness
regulates the flavor contribution in relationship to irritation. Irritation
degree and type can be either a positive or a negative attribute when
designing a product. The leaf stalk position has the best strip yield
when processing. All of these attributes makes this stalk position the
most expensive to purchase.
The tips make a positive contribution to flavor. Once again, the degree
of ripeness in this stalk position regulates the ratio of the
irritation:flavor contribution.
The flue-cured stems are important to a blender for two basic reasons:
lower blend cost and maintenance of blend nicotine content. Flue-
cured stems can be put into the product at levels of 2 to 10% without
any adverse taste characteristics. They can be puffed or expanded to
increase filling value. This, combined with total utilization of raw
material, lowers the blend cost.
The nicotine of good flue-cured stems ranges from 0.35 to 0.70%.
Sugar content is lower than in the comparable stalk position from
which they were derived. Nitrate content is in an acceptable range.
The major negative attribute to be aware of is the chloride content. If
it is greater than 1.0%, the stems can taste similar to how a wet dog
smells.
Burley
Burley tobacco is an essential ingredient in a blend, because it adds
strength and character to the smoke. Like flue-cured, burley tobacco
can be broken down into four basic stalk positions, each of which
makes its own individual contribution to smoke.
Flyings this lower stalk position is only good for use as a filler. Its
contribution to increased filling value, either expanded or
nonexpanded, must be regulated carefully, due to the irritation
potential. The burning character of this stalk position is good, due to
the higher nitrate content.
Mid-stalk this position can be used as filler or as modifier in a blend,
depending on its degree of ripeness, amount of essential oils
(subjective) and shatterability. A dull leaf surface finish, characterized
as 'boney', can have a negative effect in the smoke. The ideal character
of this stalk position should be an oily sheen and a feel of silk. Burley
from this stalk position can vary in taste characteristics by its country
of origin. This is due to cultural practices and climate more than it is
to genetics. Use in a blend can depend on the taste character the
blender is trying to achieve.
Upper middle stalk this position of burley is the basis for its strength
and flavor in a blended product. The upper middle stalk nicotine
content is the highest of all the burley stalk positions that have been
topped during the growing cycle. The color should be a rich
mahogany with oily characteristics, with no dull or 'boney' finish.
More so than with any other type of tobacco, curing is the key to a
good character from this stalk position. Stalk cut is preferred over
primed burley. The curing must be a slow decaying process to achieve
the proper chemistry and smoke character. Again, the stalk position
can have different taste

 Page 348
characteristics from one growing region to another, even within the
USA and even more so from country to country. Climate and curing
regulate this stalk position's smoke characteristics.
Tips these are another strength and flavor contributor. The nicotine
content is usually slightly lower than that of the upper mid-stalk
positions of topped burley. This selected tobacco, especially the 'red
type' with an oily sheen, is extremely flavorful. Also, this type of
tobacco comes from the upper mid-stalk position.
Unlike flue-cured, burley stems should not be used in the blend, due
to their taste character. To be of use, they must be extracted and/or
used in reconstituted tobacco.
The USA is considered a benchmark for good quality burley tobacco.
The main reason for this is the natural environmental conditions
needed for proper air curing. Other areas produce a very desirable air-
cured tobacco, but it is different from USA burley.
Oriental
Oriental tobacco possesses certain specific characteristics, such as oils
and resins in the leaf, that provide the distinct taste and aroma for
which Oriental tobacco is famous. Oriental tobacco is used in the
American blended cigarette at levels of 3 to 15%, depending on the
taste character desired.
The majority of the world supply of Oriental tobaccos are produced in
a very limited area of the Near East and Europe. Turkey and Greece
are considered the standard for good quality and former Yugoslavia
and Bulgaria produce an acceptable Oriental style. Very few other
countries produce an international quality of Oriental, due to climate
and soil conditions. Most Oriental and semi-Oriental types of
tobaccos, produced outside of their growing areas, are used for local
consumption in straight or blended products.
The major types of Oriental produced in Turkey are Ismir and
Samsun. Those produced in Greece Basma, Elassona, Katerini and
Kaba-Kulak Bulgaria and former Yugoslavia have their own specific
type of Oriental which has unique taste and aroma. It must be
emphasized that each type of Oriental tobacco has its own taste and
aroma characteristics. Depending on the cigarette marketing brief, the
blender must choose between these different Oriental or semi-Oriental
tobaccos for the specific taste required.
Cultural Practices
Agronomic/cultural practices play an important role in producing a
good quality tobacco from each type. Table 11.2 summarizes the
major cultural practices for each type. The purpose of highlighting
these agronomic/cultural practices for each tobacco type is to illustrate
that these practices produce different products. The combining of the
various types of tobacco is the art of blending to get the desired taste
character.
Marketing
Examples of basic types of tobacco marketing systems are given
below. There are variations of these three, but these represent the
basic system.
Table 11.2 Desirable agronomic practices for flue-cured, burley and
Oriental tobacco production.
Flue-cured Burley Oriental
Soil Low/Medium fertility Medium/high fertility Low
fertility
Fertilizer Low nitrogen Medium/high nitrogen None
Medium phosphorus Medium phosphorus
and potassium and potassium
Plant 13 585 to 14 820 per ha18 525 to 20 995 per ha>185 250
population per ha
Topped and Yes Yes No
suckered
Harvesting Primed Stalk cut Prime
Curing Flue (with heat) Air Sun
Time of 6 to 9 days 5 to 7 weeks 12 to 17
curing days

 Page 349
Auction system: tobaccos in the USA and several other countries are
sold to tobacco companies and dealers via an auction system.
Generally, the farmer delivers the tobacco to a 'warehouse' and all the
companies openly bid during the auction sale on each individual lot
(generally 70 to 115 kg).
Contract system: this system is utilized when companies have a
contract with a farmer to purchase his entire crop. This contract is
sometimes a 'gentleman's agreement' and not a written contract.
Bid system: the farmer will present his tobacco on a given day and the
companies will have the opportunity to place a bid on the farmer's
entire crop.
Processing and Aging
Once flue-cured and burley are purchased from the farmers, the
tobaccos are delivered to a processing plant (sometimes referred to as
a 'green leaf threshing plant'). At this facility the tobacco is sent
through a series of threshers where the lamina is removed from the
stem and packed in containers suitable for storage. Table 11.3 shows
the average yields for flue-cured and burley when processed.
Table 11.3 Flue-cured and burley
processing yields.
Parameter Flue- Burley
cured
Lamina (%) 67 63
Stem (%)
large 17 16
small 4 5
Fines (%) (<0.32 2 2
cm)
Shrinkage (%) 10 14
Common storage containers include the hogshead, a round wooden
container or the tersa bale, a rectangular bale, both of which hold 454
kg lamina or the T6B, a rectangular bale that holds 193 kg lamina.
After processing, the tobacco is aged prior to cigarette manufacturing
to enhance the smoking quality of the tobacco. The aging requirement
depends on the type and grade of the tobacco. Most flyings and sand
lugs of both burley and flue-cured do not improve with aging. From a
blending standpoint, crop year interfacing is more important than the
aging of this stalk position. To produce uniform and consistent
cigarettes, it is essential to have a minimum of 2 crop years in a
product. Sometimes 3 crop years are beneficial to blend off adverse
effects of crop year differences.
Mid-stalk and upper stalk grades greatly benefit from aging; 18
months are essential for modifier and flavor grades. Beyond 24
months, it is questionable whether the degree of improvement offsets
the capital tied up in the inventory. Once again, crop year interfacing
is important to achieve a consistent, high quality blended product. The
more adverse the individual crop year, the longer the blender will
stretch out the usage of a product.
Some countries will ferment (heat treat) flue-cured and/or burley.
There is no question that this process increases the smoke taste. This
practice is used when sufficient inventories are not available for
normal aging. Good quality flue-cured or burley tobaccos, properly
produced and cured, do not need fermentation, if sufficient inventories
can be maintained for normal crop year interfacing.
Oriental tobacco is not threshed, but goes through 'manipulation',
which is a process of separating and grading the tobacco and packing
it in a small rectangular hessian burlap bale. After manipulation, the
tobacco remains in the country of origin, going through a 'natural' (3
to 4 months) or 'artificial' (2 to 3 weeks) fermentation which is simply
a rapid aging process. Once fermented, the tobacco is shipped and is
ready for use.
Reconstituted Tobacco
Another contribution to the blend is reconstituted tobacco. There are
several types of reconstituted tobaccos, but it basically can be
categorized into two types: band cast and paper cast. Each company
has their own process for making this material, but the feed stocks are
the same: stems include both small and large burley stems and small
flue-cured stems generated from threshing. Examples of materials
used for making reconstituted sheets include:
Stem meal small particles screened out during stem expansion,
cutting and the drying process.
Factory winnowers slivers of stem recovered at the cigarette maker.
Lamina fines fines generated during green leaf threshing.
Manufacturing fines fines produced during the cigarette
manufacturing process.

 Page 350
Reclaim shorts returned trade goods or factory rejects.
Oriental scraps small particles.
As pointed out earlier, reconstituted tobacco is included in a blend at a
level of 0 to 25%. Its effect on smoke depends on the type of
reconstituted tobacco used and the level of usage.
Basic Chemical Components
The chemical difference between the three types of tobacco is best
exemplified by two major components nicotine and sugar.
Sugar is a familiar substance and most of us are aware that there are
different types of sugar, such as glucose, sucrose, etc. The types of
sugar are classified as either reducing or nonreducing. Reducing
sugars are capable of producing chemical reactions known simply as
'reduction' (i.e. glucose) and those not capable of such a reaction are
known as nonreducing (i.e. sucrose). The natural sugar in tobacco is
predominately reducing sugar.
Table 11.4 summarizes nicotine and sugar content in the three tobacco
types. Data for US flue-cured and burley are exhibited as they are
considered the standard. This table shows flue-cured contains
relatively high amounts of nicotine and sugar, burley contains a high
amount of nicotine and no sugar, and Oriental contains low levels of
nicotine and relatively high amounts of sugar.
Table 11.4 Ranges of nicotine and sugar by
tobacco type.
Reducing Total
Nicotine sugars (%) sugars
(%) (%)
Flue-cured 1.54.5 8.025.0 8.030.0
Flue-cured 24.5 8.020.0 8.025.0
(USA)
Burley 1.55.0 0 0
Burley 2.55.0 0 0
(USA)
Oriental 0.52.0 10.017.0 10.020.0

For nicotine, the relatively high amounts found in flue-cured and

burley are due to several factors such as variety selection, fertilization,
plant population, topping and sucker control. The sugar content in
flue-cured and Oriental is related to the high initial starch content and
the subsequent conversion of starch to sugar during the curing
process. Burley is relatively low in starch when harvested, yet is
eventually oxidized during the air-cured process. The relationships of
nicotine, sugar and other chemical constituents are the basis of the
unique properties of the tobacco types and their grades. For example,
the higher the nicotine, the greater the impact and strength of the
tobacco.
Cigarette Manufacturing
All types of tobacco are aged at 12.5 to 13.5% moisture content. At
this moisture content, all tobaccos have a degree of shatterability.
Therefore, the first step in cigarette manufacture is to increase the
moisture content. Flue-cured and burley are generally conditioned in a
steam vacuum chamber. Oriental, being more delicate, can be
conditioned in a high humidity room over a longer period of time.
Schematically, the flow through cigarette manufacture is as follows
(Fig. 11.1):
Burley conditioning burley blending bin
Flue-cured conditioning Flue-cured blending bin
Oriental conditioning Oriental blending bin
Individual grades within each type are blended together for
uniformity.
The blend is the single most important factor in determining the
smoke quality of a product. Blending can be defined as the combining
of raw materials in grades identifiable through visual, chemical and
smoking characteristics. Through blending, taste character,
irritation/harshness and strength can be altered to the desired product
brief. The blender must be familiar with the smoke characteristics of
each individual grade of each tobacco type. He must be able to look at
the quality of each individual grade and know how that appearance of
the tobacco will marry with other blend components and smoke in the
final product.
Extensive chemical and physical properties are essential tools to the
blender, but his taste perception is the final tool which determines
whether his blend meets the product specifications.
Casings and Flavorings
The main character of tobacco smoke is determined by the tobacco
blend used. However, this character

 Page 351

Fig. 11.1
Typical tobacco flow during cigarette manufacturing.
can be fine tuned by the use of additives to the tobacco. These
additives are generally classified as casings and flavorings. Casings
are usually applied to the tobacco strip or leaf early in the primary
processing scheme. Their function is to improve smoke quality and
the processability of the tobacco. Being used early in the process,
different casings can be applied to the various constituents of the
tobacco blend (e.g., burley, flue-cured, Oriental, etc.). Flavorings, on
the other hand, are generally applied to the total tobacco blend as one
of the last steps in processing. Their function is to improve smoke
quality and, to a lesser degree, impart a pleasant pack aroma. They are
usually more volatile than casings and are applied at minute levels
(e.g. parts per million rather than parts per hundred).
Casings are applied to tobacco for a number of reasons. First, they
tone down or mute the strength and harshness of smoke. This
ameliorating effect is particularly important on the burley portion of
the blend which otherwise imparts a great amount of strength and
irritation (i.e. the sensation of irritation). Secondly, they improve the
'processability' of the tobacco. By making the strip or leaf more
pliable, less particle size degradation occurs in subsequent processing.
This results in improved cigarette physical properties. A third, very
important effect of casing is to 'blend' the smoke irritations with the
other sensations. To a large degree, this effect is to remove the
sharpness from a given level of an irritation, making it more tolerable.
The fourth important effect of casings is to add deep flavor notes to
the smoke. These flavor notes impart a fullness or body to smoke
which further masks a given level of irritating sensations. Casing
ingredients include water, humectants, sugars, cocoa, licorice and fruit
extracts which are all available through commercial flavor houses.
Burley receives the heaviest casing load to tone down the harshness to
smoke, but it enhances the flavor. After casing, the burley must
undergo a hard redrying.
Flue-cured delivers a somewhat lighter smoke than burley, so
amelioration is not needed to the same degree. The use of humectants
is desirable.
Flavor materials are applied to the cut tobacco and are usually carried
in an alcohol base. The alcohol evaporates, leaving the flavor adhered
to the tobacco. The intent of flavoring is to enhance the tobacco taste,
yet not mask it. There are literally thousands of flavor compounds and
components. The important point of flavoring tobacco is that the
flavor material may be desirable in a bottle or flask, but it can change
when pyrolized in a cigarette. It must also enhance the pack aroma, as
well as the tobacco taste and side-stream aroma.
Subjectivity
Subjectivity is the key to a successful product, because the consumer
pays money according to his subjective taste. But subjectivity is
inherent in the entire cigarette manufacturing process. Although
chemical and physical tools are utilized, subjectivity criteria are used
by geneticists, farmers, tobacco buyers, tobacco graders, processing
personnel, blenders, cigarette manufacturers and, ultimately, the
consumer.

 Page 352
References
Akehurst. B.C. (1981) Tobacco, 2nd edn. Longman, London and New
York.
Brooks, J.E. (1953) The Mighty Leaf. Alvin Redman, London and
Sydney.
Salzman, E.I. (1963) Organoleptic investigations of the characteristics
of the tobacco leaf and cigarette smoke and scientific confirmations to
day. In: Proc 3rd World Tob. Sci. Congr., Salisbury, Rhodesia, pp.
50921.

 Page 353

11B
Cigarette Design and Materials
Alan Norman
R.J. Reynolds Tobacco Company
Winston-Salem, North Carolina, USA
Introduction
The science of cigarette design probably started with the earliest
manufactured products in the 1890s. Certainly, fledgling cigarette
manufacturers were striving to differentiate their products from those
of the competition. Most efforts centered on promoting taste
signatures from regional tobacco blends. Others focused on physical
characteristics of the products. Patents from the 1890s and early 1900s
describe perforated cigarette paper, oval shaped products and various
filters that might be attached.
By the 1930s manufactured cigarettes were the norm. Scientists
connected with the industry began to research ways to analyze
tobacco constituents and cigarette smoke. The following quote (Tilley,
1985) is a charge to a newly-hired tobacco researcher from executive
management representing direction that remains as timely today as it
was in 1934:
'In my opinion we have reached the point in the tobacco business, when
much attention should be paid to research work. With the growth of the
complexity of certain processes, and the more exacting requirements
demanded of the finished product, it is becoming increasingly true that
decisions relating to changes in materials or processes require reference to
the men in the laboratory. These men should become more dependent upon
the knowledge of the properties of materials, the discoveries of new
methods of treatment, new methods of test, and the elaboration of new
processes discovered and perfected in the laboratory. We have great hopes
that each succeeding year will find the laboratory in the lead for reducing
 manufacturing costs without sacrificing quality.'
The foundations for modern cigarette design include early attempts at
understanding how smoke is formed and analysis of the differences
between the chemical make up of tobacco and that of smoke. As
manufacturers continued their efforts to differentiate their products,
industry scientists and technologists directed their efforts to
developing product variants with appeal to broader ranges of smokers.
The cigarette design field encompasses two broad areas. One is the
continuing effort to develop detailed understanding of smoke
formation and composition through the application of the latest
analytical technology to unravel aerosol dynamics, combustion and
pyrolytic processes in burning cigarettes, and transport of the smoke
along the cigarette rod. Advances in chemical analysis techniques
allow quantitation of thousands of smoke and tobacco constituents.
Reviews of smoke formation (Jenkins & McRae, 1996) and chemical
analysis techniques applied to tobacco and smoke (Green &
Rodgman, 1996) describe these fundamental aspects of the cigarette
design discipline.
The second broad area of the field encompasses physical aspects of
cigarette design. A wide range of filters, ventilation systems, cigarette
papers and product configurations can be used for product design.
Avoiding trial and error combinations of the various choices available
requires understanding of effects of component materials on smoke
yields, resistance to draw, burn rate and other characteristics important
to product acceptability among smokers. Material choices also affect
product cost, consistency and manufacturability on high speed
cigarette makers. A review on the physical design of cigarettes
(Durocher, 1996) details the range of material and manufacturing
considerations required for developing new or modified products.
Considering the complexities involved, it is not surprising that the
cigarette design field employs a variety of specialists. Fundamental
researchers focus on smoke formation and smoke analysis. Materials
scientists concentrate on improving machinability and consistency of
papers and filters. Applied researchers work toward understanding
and modeling system performance. Tobacco blend specialists artfully
combine tobaccos for optimal taste. Product development experts
integrate available information and materials to produce new
commercial cigarette designs.

 Page 354
Three elements have provided impetus for continued interest in
cigarette design since the 1950s:
(1) Reduced smoke yields in response to demand from smokers and to
government mandates.
(2) Proliferation of product configurations to appeal to market niches
demanding length, diameter, flavoring and packaging variants.
(3) Cost and quality control.
Each of these initiatives continues to challenge the cigarette design
discipline to develop more detailed understanding of the product and
deliver innovations to meet expectations of the market place.
Reduced Smoke Yields
A few filtered cigarettes were available in the USA and in Europe as
early as the 1930s (Tilley, 1985; Deutsch, 1996). Cigarette holders
were popular enough to prompt a 1938 Reader's Digest article rating
their effectiveness (Littell, 1938). Thus, smokers' tastes had begun to
change long before the large scale manufacture of filtered cigarettes
that started in the 1950s. Popularity of filtered cigarettes skyrocketed
during the 1950s, and demand for products with lower yields of tar
and nicotine escalated. The industry responded With several cigarette
design innovations that facilitated reduced smoke yields. Figure 11.2
shows sales-weighted tar and nicotine yields from 1954 through 1994
and indicates approximate timing of USA introduction of filters,
reconstituted tobacco, high porosity paper, filter ventilation, charcoal
filters and expanded tobacco. None of these design elements could
have been used alone to achieve the reduction in tar yield
 Fig. 11.2
US sales weighted average (SWA) Federal Trade Commission (FTC) tar and
nicotine yields and selected product innovations.
shown. Rather, these design elements have become part of the product
system required to obtain lower smoke yields. Because of the
systematic approach taken by the industry to lower smoke yields,
modern filtered cigarette designs are considerably different from those
of early products (Norman, 1982). Table 11.5 shows a comparison of
a popular filtered cigarette design from 1955 with that of the same
brand today.
Table 11.5 Comparison of early and current filter
cigarette designs.
Characteristic 1955 1996
Cigarette length (mm) 85 84
Filter length (mm) 15 21
Tobacco rod length (mm) 70 63
Circumference (mm) 25.4 24.8
Tobacco weight (g) 1.0 0.8
Expanded tobacco content none 15
(%)
Paper permeability (CU) 15 24
Filter ventilation (%) none 17
Filter efficiency (%) (WTPM) 18 49
Tow item (dpf/TD) 8.0/700003.3/30000
&!;Tar' yield (mg/cig) 34 16
Nicotine yield (mg/cig) 2.7 1.1
Puff count 15 8

The current design has reduced tobacco weight through expanded

tobacco inclusion and smaller rod dimensions, a longer and more
efficient filter, filter ventilation and higher permeability paper. The
combination of these design changes resulted in tar and nicotine yields
that are less than half of those of the 1955 version. The trend to lower
sales weighted smoke yields continues today, driven by the popularity
of the low tar segment and by reduced yields in the full flavor
segment. For example, cigarettes with 15 mg tar yield or less
represented 58.2% of the USA market in 1992 and increased in share
to 62.6% in 1996 (Maxwell, 1996). Knowledge of the effects of each
of the cigarette components listed above on smoke yields, physical
characteristics, manufacturing characteristics and smoke quality is
essential for developing modern cigarette designs. Cigarette designers
must balance the use of these combined elements to produce cigarettes
with the desired smoke yields, smoking quality and economy of
manufacture. Efforts to improve effectiveness of the existing design
elements must continue. More importantly, development of new
methods for reducing overall yields and even yields of specific


 Page 355
smoke constituents must continue in order to meet smokers' demands.
Brand Proliferation
A visit to any store that retails cigarettes will confirm the wide array
of product configurations available. Almost all popular brands are
sold in various lengths, tar levels and packaging styles. Mentholated
products are available as stand-alone brands or as line extensions of
popular regular brands. The wide range of available product
configurations poses special challenges in cigarette design. Each
configuration can have different specifications for filters, papers,
ventilation levels, tobacco blend, processed tobacco content and tar
yield. Because of the logistics of dealing with such a range of
materials and product specifications, cigarette design models have
been developed to reduce trial-and-error experimentation to develop
new configurations. More recently, demand for lower-priced products
created a new wave of brand proliferation in the marketplace. In the
USA, lower-priced products include 'branded generics' (i.e. national
brands with reduced prices), private label cigarettes (i.e. retailers'
brand name and packaging) and generic products. Reduced price
cigarettes represent a cigarette design challenge in that acceptable
smoking quality must be maintained with the use of lower cost
tobaccos and materials.
Cost and Quality
As with other packaged goods, consumers demand high quality from
the cigarettes they purchase. To smokers, quality means consistent
taste from every cigarette and no functional defects such as loose
tobacco, stale product or holes in the paper. Given the speed at which
cigarettes are made (8000 to 9000 per minute), even approaching this
expected level of quality is remarkable. High quality manufacture at
speeds necessary to meet volume and cost requirements has resulted
from years of technological refinement in tobacco handling, making
and packing equipment, and advances in cigarette materials. For
example, early vented-filter designs typically used mechanically
perforated tipping and permeable plugwraps, both with relatively high
permeability variabilities which resulted in significant ventilation
variability. Since filter ventilation level strongly influences smoke
yields, variability in ventilation level will cause smoke yield (and
taste) inconsistency. Ventilation variability has been substantially
reduced by using lasers to perforate the tipping either off-line or on-
line during cigarette manufacture.
Manufacturers continually seek ways to reduce materials and tobacco
costs to keep retail prices at reasonable levels without sacrificing
product quality. Since tobacco is the most expensive component of
cigarettes, most efforts are aimed at reducing waste and increasing the
number of cigarettes made per pound of tobacco. As new tobacco
handling processes for cost control become available, cigarette design
changes may be needed to adjust smoke yields or other characteristics.
Development of more efficient, less expensive filter and paper
materials is another continuing goal. Again, as materials are modified,
cigarette design changes are often necessary to maintain product
performance. As discussed above, lower-priced cigarettes represent a
relatively new class of products that intensified cost reduction efforts
and required cigarette designs that combined the most cost effective
tobacco, paper and filter materials available.
Cigarette design is now a wide ranging discipline that allows
manufacturers the flexibility to develop reduced yield products as well
as variants for a highly segmented market and to maintain reasonable
costs while continuously improving quality. At first glance cigarettes
appear very simple in construction and design. Typical low tar
products are made with a tobacco blend that may contain a few
flavorings, cigarette paper, filter material, plug wrap, tipping paper
and adhesives. Broadly speaking, only six or seven components are
used to make any cigarette. The concept of simplicity begins to vanish
when one considers that filter type and ventilation level alone can be
'traded' over a wide range to make dozens of variants with the same
smoke yield. Realizing that many other characteristics can differ
among these constant yield designs reveals the complexity of cigarette
performance. Formation of smoke is an incredibly multifaceted
process. Considering that many design changes influence smoke
formation, one must conclude that cigarette functional complexities
belie the simplicity of their construction.
Cigarette Smoke and Smoke Formation
The physicochemical nature and mechanisms of cigarette smoke
formation have implications for many cigarette performance
characteristics including filtration processes, puff-wise smoke yield
variation and

 Page 356
smoke composition. The following brief summary of smoke
characteristics and formation is offered here as a foundation for
discussions of cigarette design elements later in this chapter. Smoke
formation is a complex and intriguing topic well covered in the
literature. Detailed information on smoke formation is available in
published reviews and references therein (Johnson, 1977; Baker 1980;
Baker; 1984; Baker, 1987; Baker & Robinson, 1990; McRae, 1990;
Jenkins & McRae, 1996).
Cigarette smoke is an aerosol. That is, smoke is a two-phase system
comprised of condensed phase particulate matter suspended in a gas.
The gaseous portion of cigarette smoke contains hundreds of chemical
constituents including fixed gases (N2, CO2, CO) and traces of low
molecular weight vapors. Smoke particles are more chemically
complex and contain thousands of both volatile and nonvolatile
compounds.
Figure 11.3 shows the approximate mass composition of mainstream
smoke gas and particulate phases categorized by classes of constituents
(Green & Rodgman, 1996). In this example, the particulate phase
represents about 4.5% of the whole aerosol while the total gas phase
constitutes the remainder. Most of the gas phase is nitrogen and
oxygen, but about 23% is represented by gases produced from burning
the tobacco.
 Fig. 11.3
Whole cigarette smoke composition by chemical class (Green & Rodgman, 1996).
With a particulate mass concentration of around 4 to 5%, cigarette
smoke is a very high particle concentration aerosol. Cigarette smoke
particles are in the submicron size range, which is expected for
condensation type aerosols. Table 11.6 shows properties of mainstream
and sidestream smoke from several cigarette types (Ingebrethsen,
1986). Because of its high particulate concentration, cigarette smoke
particles coagulate rapidly and increase in average size with aging.
These dynamics are important in considerations of mainstream smoke
formation.
During a puff, air enters the lit end of a cigarette just in front of the
paper char line between the paper and the coal. Near the point of air
intake, temperatures at the periphery and at the center of the coal can
exceed 900°C. As the air enters the cigarette, it is heated and thermally
degrades the tobacco. Both combustion and pyrolysis processes occur
during the degradation. Gas in and behind the coal region is
significantly depleted in oxygen and enriched in hydrogen, indicating
pyrolysis products are formed in the coal and the region very near it in
a chemically reducing atmosphere. The thermal degradation produces a
concentrated vapor just behind the coal. The vapor also contains
nonvolatile organic and inorganic compounds evolved from destruction
of the tobacco's physical structure. As this 'cloud' travels down the
tobacco rod, it rapidly cools and the volatile materials condense on the
unburned tobacco and form aerosol particles. The aerosol particles are
deposited to some degree by filtration as the smoke travels through the
rod. The portion of aerosol that is not removed via condensation or
filtration along the tobacco rod ultimately exits the cigarette mouthend
as mainstream smoke.
Cigarette Testing
A few standardized tests are integral to cigarette design. These include
mainstream smoke yields; pressure drops of the filter, tobacco rod and
finished cigarette; permeability of the papers used in construction;
ventilation level; and firmness of the filter and tobacco rod. These tests
relate to factors influencing smokers' acceptance (smoke yields, draw,
firmness) and provide indications of the design approach used (filter
pressure drop, ventilation type and level). A complete cigarette
specification is much more extensive than the tests described here.
Typically, cigarette paper tipping and plugwrap specifications, filter
denier, plasticizer type and level, various dimensions, adhesive types,
tobacco lamina and processed tobacco levels and flavorings would be
included. Each of these items is important to consistent manufacture of
quality products and can influence final product performance.
(a)
'Tar' and Nicotine Yields
The procedure for measuring mainstream smoke yields of `tar' and
nicotine is standardized so that results from

 Page 357
Table 11.6 Cigarette smoke properties (Ingebrethsen, 1986).
( )a Mainstreamb ( )a Sidestreamc
CigaretteGeometric Logarithmic Number Geometric Logarithmic
mean Mode standard concentration mean Mode standard
diameter diameter deviation N (1010 cm- diameter diameter deviation
(µm) (µm) 3) (µm) (µm)
A 0.167 0.140 0.42 2.3 0.108 0.090 0.41
(0.003) (0.01) (0.2) (0.010) (0.04)
B 0.167 0.141 0.41 3.1 0.104 0.088 0.41
(0.003) (0.01) (0.2) (0.009) (0.04)
C 0.163 0.135 0.46 3.5 0.095 0.077 0.46
(0.002) (0.01) (0.3) (0.006) (0.03)
D 0.179 0.153 0.40 3.0 0.118 0.105 0.34
(0.003) (0.02) (0.5) (0.005) (0.02)
E 0.177 0.152 0.39 2.8 0.116 0.103 0.35
(0.004) (0.02) (0.4) (0.008) (0.04)
a The numbers in parentheses show standard deviation.
b Obtained from the fourth puff.
c Obtained during the interval between the second and third puff.
A Flue-cured
B Japanese Native, Bicyu
C Burley
D Blended
E Blended

various testing laboratories are comparable. Two slightly different

standards exist: that of the US Federal Trade Commission (FTC) (Ogg,
al., 1962; Pillsbury, et al., 1969) and that of the International Standards
Organization (ISO) (ISO, 1991). Most of the differences between the FTC
and ISO methods are in the details of cigarette handling, conditioning and
smoke collection. The puff volume, duration and interval are common to
both standards: 35 ml puffs of 2 seconds duration taken once per minute.
The flow profile during the puff is 'bell-shaped', with an average flow rate
of 17.5 ml/s (i.e., 35 ml/2 s) and peak flow rate at 1 second into the puff of
about 27 ml/s.
about 27 ml/s.
Smoking machines usually include a switch that stop the flow during
puffing when the predetermined butt length is reached. Smoking
procedures specify the butt length remaining at the end of smoking. For
filter cigarettes, the butt length is usually 3 mm past the tipping overwrap.
Butt length for unfiltered cigarettes is normally 23 mm. With the
predetermined butt length and periodic puff/smolder cycle described,
about 20 to 25% of the tobacco is consumed during puffing for typical 85
mm low tar cigarettes.
The particulate fraction of smoke collected on the Cambridge filter pad is
called wet total particulate matter (WTPM); WTPM is taken as the weight
gain of the pad during the smoking process. Materials on the Cambridge
pads are extracted in methanol and analysed by gas chromatography for
nicotine and water content. 'Tar' is the term applied to the smoke
particulate fraction minus the water and nicotine content. It is also called
'dry particulate matter' or DPM.
As might be expected, any changes in the details of standard smoking
procedures will influence measured smoke yields. For example, increasing
the puff volume, decreasing the interval between puffs or decreasing the
butt length will each result in increased tobacco consumption during the
puffing cycles, thereby increasing smoke yields.
b
Pressure Drop
Pressure drop measurements of cigarettes and their components are very
useful for estimating various product performance aspects. Filter pressure
drop is directly related to smoke removal efficiency. Tobacco rod pressure
drop indicates the packing density (or weight per unit rod volume).
Pressure drop of finished cigarettes is related to smokers' acceptance of the
product and can be used to estimate ventilation level. Component and
finished product pressure drops are routinely monitored for quality
determinations.
Pressure drop measurement entails drawing a steady flow of 17.5 ml/s
through the article of interest (filter,

 Page 358
tobacco rod or cigarette) with a manometer connected at the exit point
of the drawing flow. The manometer registers the difference between
atmospheric pressure and the reduced pressure inside the article being
measured. Essentially, the measurement relates to the power required
to move air through the article. Thus, pressure drop is sometimes
called 'resistance to draw' (RTD). CORESTA has published
recommendations (CORESTA Recommended Method No 41, 1995)
for measuring pressure drop. Among other items, the standards
include direction of flow (i.e. drawing through the open end),
preferred flow rate (17.5 ml/s; the average flow during the standard 35
ml puff of 2 seconds duration) and units of measurements (cm or mm
of water).
c
Paper Permeability
Air permeability is an essential characteristic of cigarette paper.
Permeability greatly influences static burn rate and the degree of
ventilation through the tobacco rod. For vented-filter designs using
pre-perforated tipping, plugwrap and tipping permeability determine
the degree of filter ventilation. Air permeability for each of these
materials is critical to smoke yield and cigarette pressure drop.
The permeability test involves measuring the air flow rate resulting
from applying a known pressure difference across a specified area of
paper sample. The CORESTA recommended method No 40 (1994)
defines a 2.000 cm2 rectangular test area with one side of the
rectangle of 2 cm length. This paper sample area accommodates
cigarette and plugwrap papers, which can be inherently porous or
randomly perforated, and tipping papers, which are usually perforated
in rows or narrow zones. The recommended pressure drop applied to
the test piece is 1.0 kPa (10 cm H2O). The resulting flow rate (units of
cm3/min) is divided by the measurement area to yield air permeability
in 'CORESTA units' (CU; cm/ min).
For some papers, air permeability is not linearly related to the pressure
drop across the paper. Nonlinear behavior is typical for perforated
papers and can be found in high permeability plugwraps and cigarette
papers that are not perforated. Nonlinear flow/ pressure drop
relationships influence certain aspects of finished product
performance. Therefore, characterization of the flow/pressure drop
relationship is necessary to adequately specify the paper material. In
the CORESTA recommended test method, flow rate measurements are
made at each of two pressure differentials across the test material.
These data are used to calculate the ratio:
Y = (flow at pressure A)/(flow at pressure B) × (pressure B/pressure
A)
If the resulting ratio is 1.00 (± 2%), the flow/pressure drop
relationship is assumed to be linear. Note that this ratio is sometimes
included in paper material specifications along with the air
permeability.
d
Ventilation
Ventilation is an important determinant of smoke yield and finished
cigarette pressure drop. In the CORESTA recommended method No 6
(1983), ventilation is defined as air entering any point of an unlit
cigarette other than through its lighting end. Filter ventilation is
defined as the amount of air entering the cigarette through the portion
of tipping paper that is not covered by the test holder and not
overlapping the tobacco rod. Paper ventilation is the air entering the
whole length of the tobacco rod. Total ventilation is the sum of the
filter and paper ventilation levels. Normally, ventilation is expressed
as the degree of ventilation, i.e. the ratio of ventilation air flow to the
total flow exiting the cigarette mouthend, expressed as a percentage.
Ventilation measurement apparatus includes a device that grips the
mouthend of the cigarette, providing a leak-free seal and a constant
drawing flow of 17.5 ml/s. To measure filter ventilation, a chamber
enclosing the exposed part of the filter is connected to a flow meter
that indicates the flow rate into the tipping. Total ventilation is
measured in the same way, but with the entire cigarette enclosed in the
chamber except for the portion of tobacco rod covered by the seal at
the lighting end. Any of several types of flow measuring equipment
can be used as long as the pressure drop across the apparatus is
sufficiently low (< 0.1 mbar at 17.5 ml/s) to avoid influencing the
ventilation flow. Paper ventilation is the total ventilation minus filter
ventilation. It can be measured directly as described above by
enclosing only the tobacco rod in the ventilation flow chamber.
e
Firmness
Firmness measurements are made on filters and tobacco rods to
indicate their resilience. Subjectively, cigarettes must have sufficient
firmness to impart a feeling of substance and bulk. Usually, filters and
tobacco rods are manufactured to a firmness specification. Firmness
increases with weight of filter material

 Page 359
or tobacco used. Therefore, making cigarettes of adequate firmness
with low weights of materials is economically important.
Currently, there are no published standards for firmness measurement.
Usually, firmness is measured as the displacement of a weighted
presser foot placed on the rod. The displacement is expressed either
directly or as a percentage of the rod diameter. Firmness expressed as
percentage of the rod diameter is also called 'hardness'. Measurement
results depend on the size of the presser foot, the weight applied to the
presser and the amount of time the presser is applied to the rod. In
addition, the moisture content of the test pieces will influence
firmness readings. Each of these items must be controlled to obtain
reproducible results.
Cigarette Paper
Cigarette paper probably influences overall product performance more
than any other nontobacco component. In a typical 85 mm low tar
product design, the cigarette paper represents about 5% of the total
tobacco rod weight. Despite its small contribution to the rod weight,
the paper influences the static smolder rate, ventilation, puff count,
tobacco rod (and thus the cigarette) pressure drop and yields of both
sidestream and mainstream smoke. The paper substantially determines
finished product appearance, ideally providing an opaque, wrinkle-
free tobacco rod exterior with markings or coloration specified by the
designer. High speed cigarette manufacture demands paper strength
adequate to withstand pulling the paper through the maker without
breaking or excessive stretching. Although typically porous, the paper
must not wick the seam adhesive during rod making. Because of its
importance to product appearance, manufacturing, economics and
performance, cigarette paper is a highly engineered product that
requires careful specification to achieve the desired results.
a
Paper Composition and Specification
Cigarette paper is composed of an inorganic filler and cellulosic fiber.
The most common inorganic filler is calcium carbonate (CaCO3).
Special fillers may be used for reducing sidestream smoke yields.
Common fibers used for cigarette papers include flax and wood pulp.
Papers are made by casting a dilute slurry of filler and bleached fiber
on a moving, porous belt that allows water to drain from the slurry to
form the finished sheet. Most cigarette papers are white. Since no
whiteners are added during manufacture, the filler and fiber bleaching
determine the whiteness of the finished paper. The filler holds the
fibers apart, creating pores within the paper structure. Air
permeability is a key paper specification governing smoke yield that
results from the amount and size of the filler particles, the size of the
fiber particles and the density (basis weight) of the sheet. Paper can be
perforated to increase permeability beyond that created by 'inherent'
pores formed during manufacture. Methods of perforation include
electrostatic, mechanical or laser. Perforation increases measured air
permeability and ventilation through the paper on finished cigarettes.
However, unlike permeability provided by pores, perforation does not
influence static burn rate. Ranges for common paper specifications
such as permeability, filler content, basis weight and thickness
(caliper) are shown in Table 11.7.
Table 11.7 Commercial cigarette paper
specifications and ranges.
Parameter Range
Permeability, 10120 from inherent
CU pores, up to 200 perforated
(cm/min at 1
kPa)
Filler content 2040 (30 typical)
(w/w, %)
Basis weight 2230 (25 typical)
(g/m2)
Thickness 0.030.05 (0.039 typical)
(caliper) (mm)
Opacity (%) 7082 (77 typical)
Tensile 2.54.5 (3.5 typical)
strength (kg/3
cm)
Whiteness 8688 (flax fiber)
(brightness)
(%)
9294 (wood pulp)

Burn modifiers or ash conditioners, typically alkali metal salts of

organic acids, can be applied to the sheet after formation. Levels of
these additives are typically between 0.5% and 3% of the paper
weight. Because burn modifiers impart characteristic tastes and
aromas to the smoke, regional preferences have developed for certain
types of ash conditioners. For example, sodium/potassium citrate is
primarily used in American blend cigarettes, acetate in Germany and
tartrate in Asia (Baskevitch, 1986). The alkali metal salts provide a
whiter ash and increase static burn rates (i.e. linear burn rate of the
smoldering tobacco rod). Other additives (e.g. monoammonium
phosphate) condition the ash but do not alter static burn rates (Owens,
1978; Townsend, 1984). Table 11.8 shows typical cigarette paper
additives, levels and their functions.

 Page 360
Table 11.8 Cigarette paper additives and
typical levels (Owens, 1978).
Functional category
Increase Ash conditioners Decrease
static burn static burn
rate rate
Alkali metal Monoammonium Aluminum
citrates phosphate (MAP) chloride
0.51.5% up to 3%
Alkali metal Urea NaVO3
tartrates
0.51.5%
Alkali metal Na2B4O7
acetates
0.51.5%
Alkali metal Na2MoO4
malates
Alkali metal
formates
Alkali metal
phosphates
Alkali metal
nitrates
Alkali metal
sulfates
Alkali metal
carbonates
Alkali metal
halides

b
Paper Effects on Smoke Yields
Cigarette paper air permeability provided by inherent pores within the
paper influences static burn rate, ventilation through the paper and
outward diffusion of low molecular weight gases formed during puffing.
Yields of all smoke constituents are reduced with increased paper
permeability due to the combination of these effects. However, the degree
of reduction varies with smoke constituent class, as shown in Fig. 11.4
(Owens, 1978).
For inherently porous papers, ventilation through the paper is directly
proportional to the permeability (Mattina & Selke, 1977; Selke &
Matthews, 1978). The following relationship exists between paper
ventilation and permeability (Baker & Robinson, 1987):

where
Qp = flow through paper (ml/min) given flow Qd at the mouthend
P = tobacco rod pressure drop measured at flow Qd (cm H2O)
 Fig. 11.4
Reduction of smoke yields with inherently permeable cigarette paper (Owens, 1978).
r = tobacco rod radius (cm)
w = paper permeability/10 cm H2O (cm/min/cm H2O)
L = tobacco rod length (cm)
Vp = ventilation through the paper
It also includes a dependence of paper ventilation on the tobacco rod
pressure drop which is analogous to the 'upstream' pressure drop effect on
filter ventilation shown elsewhere in this chapter. That is, flow through
the paper increases with increased tobacco rod pressure drop. Equations
11.1 and 11.2 also demonstrate that ventilation through the paper will
decrease substantially as the tobacco rod length becomes shorter during
smoking (Baker, 1984).
As with filter ventilation, the puff volume taken at the coal is reduced
when ventilation via the cigarette paper is increased. Since less tobacco is
consumed due to the coal puff volume reduction, smoke yields are
reduced. Yield reduction of low molecular weight gases (e.g. CO and
CO2) is nearly linearly related to the paper permeability. This indicates
that 'fixed' gas yields are primarily influenced by the ventilation level. A
secondary effect on fixed gas yield is outward diffusion through the paper
(Owen & Reynolds, 1967; Morie, 1976; Muramatsu, et al., 1977;
Durocher, et al., 1978). The extent of CO and CO2 diffusion is
proportional to the permeability and the smoke velocity (Baker, 1982). In
cigarettes with vented filters, diffusion is an

 Page 361
important determinant of CO yield. At high filter ventilation levels, as
much as 90% of the CO produced at the coal is lost through the
wrapper (Townsend & Norman, 1982). Diffusion of CO in this case is
facilitated by the lower smoke velocity in the tobacco rod resulting
from high filter ventilation.
Another secondary effect is produced from increased burn rate due to
higher permeability. Higher burn rates during the inter-puff smolder
intervals result in relatively less tobacco available for consumption
during puffing. The net effect of increased burn rate is a reduction in
puff count which, in turn, further reduces yields of all mainstream
smoke constituents.
Ventilation through the paper reduces particulate yields via coal puff
volume reduction. However, increased burn rates for higher
permeability papers result in shorter lengths of tobacco rod remaining
at a given puff number, creating a compensatory effect on particulate
yield reduction. Significant particulate matter deposits on the
unburned tobacco behind the burning zone during each puff. The yield
for a given puff can be described by (Baker, 1984) by:

where
Ci = yield of constituent for puff i
Gi = amount of constituent formed in the burning zone
µ = condensation/filtration coefficient
fi = length of tobacco column remaining after puff i.
As permeability increases, the tobacco rod is shorter at a given puff
number because of the increased static burn rate. Due to the shorter
rod length, there is less removal of particulates via filtration by the
tobacco rod. Condensation is essentially independent of the length of
unburned tobacco rod, apparently taking place within about 10 mm
behind the char line (Townsend, 1983). As a consequence, particulate
yield reduction levels off at very high paper permeability. Particulate
smoke removal by the unburned portion of the tobacco rod contributes
in a second way to the increase in per puff smoke yields with puff
number (Townsend, 1983). During a puff, heated air entering behind
the coal produces thermal degradation products of the tobacco which
condense on the unburned tobacco behind the coal. In subsequent
puffs, some of the previously deposited material volatilizes and is
entrained in the mainstream smoke. Semi-volatile smoke constituents
(e.g. nicotine) are enriched in the smoke owing to this
condensation/re-uptake phenomenon. During later puffs, there is less
unburned tobacco for condensation. As a result, smoke yields increase
even further on the final puffs.
Perforated cigarette papers exhibit the same ventilation dependent
smoke reduction mechanisms as described above. However,
perforation does not influence burn rate. Burn rate is governed by
oxygen transport to the coal region via diffusion and is dependent on
the number of pores present in the paper (Hampl, et al., 1987).
Perforating the paper increases the air flow capability of the paper (i.e.
its air permeability), but perforations do not increase the oxygen
diffusion capability. Thus, a perforated paper has higher permeability,
resulting in a higher ventilation level than afforded by the base paper,
but static burn rate remains the same as for the base paper.
Figure 11.5 compares the effects of inherent and perforated
permeability on static burn rates and puff counts (Baskevitch, 1986).
The horizontal line in Fig. 11.5(a) indicates static burn rates resulting
from perforating a base paper with inherent permeability indicated by
the intersection point of the straight line with the curved line showing
the effect of inherent permeability. Figure 11.5(b) shows effects of
inherent and perforated permeability on puff count. As discussed
above, increased inherent permeability increases static burn rate,
resulting in fewer puffs. Perforating the paper does not change the
static burn rate. Higher paper ventilation levels resulting from the
perforations reduce the amount of tobacco burned during puffing,
causing increased puff counts.
For a given permeability, ventilation through perforated paper is
slightly higher than that afforded by a paper with the same inherent
permeability. This is a consequence of different pressure/flow
relationships for perforated and inherently porous papers. In
perforated papers, the perforations are relatively large in relation to
the paper thickness. Because of the perforation size, part of the flow
through perforated papers is due to inertial forces, and the
pressure/flow relationship for these papers is non-linear. Flow through
cigarette paper can be described by the following equation (Baker,
1989):

where:
Q = air flow through the paper (cm3/min)
A = area of the paper through which air flows (cm2)
P = pressure drop across the paper (cm H2O)

 Page 362

Fig. 11.5
Burn rate and puff count variation with inherently permeable and perforated
cigarette papers (Baskevitch, 1986).
Z = permeability due to viscous flow (cm/min/10 cm H2O)
Z¢ = permeability due to inertial flow
n = constant for a given paper ranging from 0.5 to 1.0.
For inherently permeable papers, the inertial term (Z¢APn) of equation
11.4 is zero. This term is appreciable for papers with large
perforations, and such papers show a nonlinear pressure/flow
relationship.
Since perforation increases the paper ventilation and not the static
burn rate, perforated paper permeability shows different effects on
smoke yields than those of inherent permeability. Figure 11.6 shows
the effects of perforated permeability on puff count and on reduc-

Fig. 11.6
Reduction of smoke yields with perforated cigarette
paper (Owens, 1978).
tions of various smoke constituents (Owens, 1978). Perforated
permeability causes a slight increase in puff count over the range
shown in Fig. 11.6 due to higher paper ventilation resulting in less
tobacco burned during puffing and the absence of perforation effects
on static burn rate. This is the opposite of the effect of inherent
permeability (Fig. 11.4). Particulate yields are reduced to a lesser
extent than the fixed gases with perforated papers. Again, this
behavior is opposite from the effect shown for inherent permeability
because perforations influence ventilation through the paper and not
the static burn rate. In fact, smoke yield reduction via perforated paper
closely resembles trends found with filter ventilation.
Any of several chemical treatments are added to cigarette papers to
alter the static burn rate of the tobacco rod or to enhance the ash
appearance. Alkali metal salts of organic acids are typical treatments
to increase static burn rate, with sodium or potassium citrate, tartrate
and acetate the most commonly used. Levels of these treatments
typically range from 0.5% to about 3% by weight of the finished
paper. Monoammonium phosphate (MAP) is applied as an ash
conditioner at around 0.5% to 1% by weight. This additive does not
alter static burn rate. Table 11.9 shows that smoke yields are
influenced by both additive types (Owens, 1978). For MAP, yields of
tar and CO increased although no change in burn rate was

 Page 363
Table 11.9 Effects of type and amount of
cigarette paper additive (Owens, 1978).
Paper Static burn Puff
additivea rate (mg/min) count ¢Tar¢ CO
(%, type) (mg/cig) (mg/cig)
None 54 9.0 26.3 12.5
0.5 52 9.1 27.2 15.2
Phosphate
1.0 51 9.1 27.7 16.0
Phosphate
0.5 62 8.8 25.5 13.4
Citrate
1.0 65 8.6 25.0 13.0
Citrate
2.0 72 8.2 24.3 12.6
Citrate
3.0 79 7.8 23.8 12.4
Citrate
a 47 CORESTA permeability paper.

observed. 'Tar' yields decreased with citrate application, as expected

from increased burn rates. However, CO yields increased at low
citrate application levels despite the increased burn rates. These
results illustrate effects of chemical treatments that are independent of
those expected for burn rate increases resulting from increased paper
permeability.
Chemical treatments lower the temperature at which thermal
degradation of the paper begins (Owens, 1978; Townsend, 1984).
Since permeability increases rapidly near the char line during thermal
degradation, increased oxygen transport to the burning zone is
expected as a result of chemical treatment. This explains faster static
burn rates and increased carbon oxide evolution for alkali metal salt
treatments. For other additives, such as MAP, the increased oxygen
availability may be offset by decomposition products of the additive,
leading to a smaller effect on static burn rates (Townsend, 1984).
Since sidestream smoke (i.e. any smoke not exiting the cigarette
mouthend) primarily results from the smolder between puffs, most of
the paper formulation adjustments discussed above influence
sidestream smoke yields. In general, paper properties that retard the
permeability increase during paper degradation will reduce sidestream
yield. Such variables include low inherent permeability (less than 15
CU), high levels of chemical additives (>3%), high basis weight or
special filler material. For example, magnesium hydroxide filler used
with other paper design components can provide reduced sidestream
tar yields (Dixit, et al., 1991).
The sidestream plume is formed from vapors exiting the cigarette just
behind the char line where paper permeability is increasing as the
paper thermally degrades (Baker & Robinson, 1990). Very low per
meability papers (i.e. less than 10 CU) show substantial reductions to
sidestream smoke yields because the permeability is maintained at a
low level as the papers begin charring. Such papers only marginally
support static smolder. Because burn rates are so low, vapors from the
burning zone condense near the char line. Higher levels of condensed
materials around the char line may also impede the sidestream plume
formation. The filler material amount and type can reduce sidestream
yields. Magnesium hydroxide is an alternate paper filler that results in
substantial sidestream smoke reductions (Hampl, 1989; Dixit, et al.,
1991). Increasing the basis weight also results in sidestream
reductions. Filler surface area has been shown to be related to
sidestream smoke yield (Hampl, 1989). Vapors may deposit on the
filler surface before diffusing out of the rod, reducing the amount of
material available for sidestream plume formation. This explanation
would account for reductions observed for both filler type and filler
amount. High levels of chemical treatment are believed to fuse the
ash, preventing formation of fissures near the char line through which
vapors can escape (Hampl, et al., 1989).
Cigarette Filters
Large-scale manufacture of filtered cigarettes began in the early 1950s
in response to demand for lower smoke yields. Filtered cigarettes
gained popularity rapidly, and today the majority of cigarettes sold are
filtered. Most cigarette filters are made from cellulose acetate and can
reduce 'tar' and nicotine yields by 40 to 50% relative to unfiltered
cigarettes. Filters within a similar smoke removal efficiency range are
also manufactured from paper. Special filter designs that include
charcoal provide selective removal of a range of vapor phase smoke
constituents. Because filter materials influence yields of some smoke
constituents to varying degrees, cigarette taste changes with the filter
material used. For this reason, choice of filter material has become
somewhat regionalized. In the USA, cellulose acetate is used almost
exclusively, while charcoal filters are very popular in Japan.
The primary purpose of cigarette filters is reduction of particulate
smoke yields, but filters also provide some cosmetic functions. Filters
serve as mouthpieces that keep tobacco particles out of smokers'
mouths and diminish cigarettes' tendency to 'lip-stick'. In this capacity,
filters are designed to have a pleasing degree of resilience and
substance (i.e. firmness) when handled by smokers. Acceptable filter
designs show

 Page 364
adequate firmness even during smoking, with minimal loss of
firmness in the last puffs from the cigarette. The filter tipping paper is
typically printed with some kind of brand identification or other visual
enhancement. These include a printed, simulated cork pattern, a white
background with colored logo or any of several possible colors.
a
Filter Materials and Specification
The primary filter design considerations are smoke removal efficiency
and the pressure drop required for the desired finished product
performance. Filter efficiency (E) for any smoke constituent is defined
as:

where Wf is the weight of smoke constituent retained on the filter and

Wr is the yield from the tobacco rod.
Typically, design targets are expressed in terms of WTPM or 'tar'
removal efficiency. Removal of specific constituents of the particulate
phase is generally in proportion to the 'tar' removal efficiency, but can
vary somewhat depending on the filter material. Adding charcoal
influences retention of some vapor phase constituents without
significantly increasing particulate removal efficiency. Cellulose
acetate and paper filter materials have only small influences on vapor
phase retention.
Filter pressure drop is a significant design consideration, as smoke
removal efficiency is directly related to it. Also, filter pressure drop is
a component of finished cigarette 'draw resistance', which is a
determinant of smokers' acceptance. That is, if the pressure drop is too
high, smokers find the excessive resistance to draw unacceptable.
Because of this, there are practical limits to 'tar' and nicotine removal
efficiency that can be achieved in marketable filters. Vented-filter
designs are used to reduce 'tar' yields below about 15 mg/cigarette,
partly in order to maintain draw resistance within acceptable limits.
Cellulose Acetate Filters
Cigarette filters are formed from a band of mono-filament (single
fiber) cellulose acetate tow. Filament diameter is about 20 microns
and a band can contain around 15 000 individual filaments. Tow is
specified by its denier per filament (dpf) and by its total denier (TD).
Denier per filament is the weight (in grams) of a single filament 9000
m in length. Total denier is the weight of a 9000 m length of the tow
band. The range of dpf used in commercial filters is from about 1.8 to
5 while TD ranges from about 20 000 to 50 000. Total denier divided
by denier per filament gives the number of filaments in the tow band.
Early filters made with round filaments of high total denier showed
only modest smoke removal efficiency. Filaments can now be
extruded in several cross-section shapes, including round, Y, I and X.
The other cross-sections were developed to increase the total fiber
surface area within a filter, yielding higher smoke removal efficiency
and pressure drop for a given weight of filter tow.
Filter tow is supplied as a continuous band of filaments of specified
dpf and TD packed in large bales. Filter making equipment pulls the
tow band from the bale and 'blooms' or opens the tow to reduce
packing density as required for target firmness and pressure drop.
Opening is accomplished by alternately stretching and relaxing
tension on the tow band as it is pulled through the filter maker. A
small amount of plasticizer (usually triacetin) is added during the
opening process to bond some of the fibers together and increase
firmness of the finished filters. Plasticizer application typically ranges
up to about 6% by weight. After opening, the tow is pulled through a
garniture of appropriate diameter and wrapped with the specified
plugwrap paper. The wrapped tow is cut into filter rods either four or
six times the tip length to be used on finished cigarettes.
Any filter tow (dpf/TD combination) can be used to manufacture
filters within a range of pressure drop values (Curran, 1976). This
range is called the tow capability. Adjusting the degree of blooming
and weight of tow packed into the filter rod provides the pressure drop
range. Filter pressure drop can be adjusted by about ± 10% of the
capability midpoint. Thus, several tow items can provide the same
pressure drop and similar smoke removal efficiency through weight
adjustment during manufacture. Often, several items also provide
adequate filter firmness. In these cases, economics drive filter
specification. Intermediate dpf and TD items used at lower weight are
usually less expensive.
Smoke removal efficiency and filter pressure drop are functions of the
total surface area of the filter fibers (Keith, 1975; Keith 1978). Total
surface area is determined by the weight (packing density) of the
fibers in the filter, the number of fibers, fiber diameter and cross-
section. To a reasonable approximation, equal length filters with the
same pressure drop show similar smoke removal efficiency. However,
reducing dpf at constant pressure drop will increase smoke removal
efficiency due to the increased surface area presented by the smaller
diameter fibers. Changing

 Page 365
from round to 'X' cross-section fibers will increase efficiency for the
same reason. Table 11.10 summarizes the effects of filter design
parameters on filtration efficiency and pressure drop.
Table 11.10 Filter parameter effects
on filtration efficiency and pressure
drop.
Design parameter Effect
Increased tow weight Increase
Increased length Increase
Increased total denier (TD) Increase
Increased denier per filamentDecrease
(dpf)
Increased filter diameter Decrease

Dual and triple filters are made by combining filter segments of

different characteristics. For example, cellulose acetate and paper
elements can be used in the same filter, or charcoal can be included in
one of the segments. Dual cellulose acetate filters can be used to
adjust the finished cigarette pressure drop in vented-filter designs
(Townsend, 1982). Manufacture of segmented filters requires
specialized equipment to combine the filter sections. Combiner
equipment accepts filter rods manufactured by standard filter making
machinery, cuts the rods into specified segment lengths and joins the
segments. A second plug-wrap is applied by the combiner to hold the
segments together. Finished segmented filter rod lengths are multiples
of four or six times the required cigarette tip lengths to ensure
compatibility with cigarette making equipment.
Charcoal Filters
Charcoal filter designs are typically multiple filter segments with one
of the segments containing charcoal granules. Two common designs,
shown in Fig. 11.7, are the dual and triple segment configurations. In
both designs, the mouthend segment is usually cellulose acetate. Dual
element filters include a second segment in which charcoal granules
are dispersed within the filter material, usually cellulose acetate. Paper
elements for dual filters are also manufactured. In this case, the
charcoal is usually bonded to the paper surface, but it can be
incorporated in the manufacture of the paper material (Blakley, et al.,
1994). Most triple segment filters include a cavity filled with charcoal
granules with cellulose acetate plugs at either end of the cavity.
Bonded elements can be used instead of the cavity.

Fig. 11.7
Dual and triple filter configurations.
Other filter materials, such as paper, can be placed at either end of the
charcoal element.
Segmented filter technology gives product designers very broad
options. Charcoal is available with a range of activity characterized by
its surface area and pore volume. Higher activity charcoal results in
increased retention of vapor phase smoke constituents. The amount of
charcoal used (up to about 60 mg per filter tip) influences retention. In
addition to a wide range of additive options, the filter element length,
pressure drop and particulate removal efficiency can be adjusted for
each segment.
b
Filter Effects on Cigarette Performance
The primary effect of fibrous cigarette filters (cellulose acetate or
paper) is removal of particulates (i.e. WTPM) from the smoke.
Particulate removal occurs largely through mechanical filtration of the
aerosol particles. Fiber filters have very little effect on the gas phase
of the smoke. Some of the vapor phase and condensable or 'semi-
volatile' smoke constituents that are primarily present in the
particulate phase are selectively retained by fiber filters. That is,
because of their affinity for the filter medium, these constituents are
retained to a greater extent than the particulate matter. Other semi-
volatile constituents elute from the filter. That is, these constituents
are retained to a lesser degree than the particulate matter removal
efficiency. Charcoal filters show much stronger selectivity than fiber
filters, especially for volatile smoke constituents.
The major mechanism of smoke retention in fibrous cigarette filters is
mechanical capture of aerosol particles. Mechanisms of particle
capture depend on the

 Page 366
size and mass of the particles and the characteristics of the fibers used
in filters. Because the concentration of particles in the smoke aerosol
is very high (Table 11.6), a high probability of collisions between
particles and filter fibers might be expected. Lower retention by the
filter might be expected from considering that particle residence time
inside cigarette filters is only around 0.04 s and particles are very
small (0.2 µ ) relative to the size of the filter fibers (20 µ) (Morie,
1977). Neither of these expectations fully describe filter performance
(i.e. cigarette filters show moderate removal efficiencies), but physical
processes that account for these characteristics have been used to
model filter performance (Keith & Derrick, 1965; Keith, 1975; Keith,
1978; Dwyer, 1986; Eaker, 1990). One example (Keith, 1978) that
describes filter efficiency and pressure drop is shown in Figs 11.8 and
11.9.

where ef =single fiber filtration efficiency

(unitless)

(µ2/s)

(unitless)

rp= particle radius (µ)

a = effective fiber radius (µ)
a = fiber volume fraction (unitless)
V0 =linear flow rate (cm/s)
Cf = Cunningham slip flow correction factor
(unitless)
µ = gas viscosity (poise)
k = Boltzman constant (dyne cm/deg)
T = absolute temp (°K)
r = polymer density (g/cm3)

Fig. 11.8
Single-fiber filter efficiency model (Keith, 1978).
Smoke particles can be captured via inertial impaction, diffusion or
direct interception. These processes can be summarized by the
following relationship (Overton, 1973):

where
C = concentration of smoke particles exiting the filter
C° = concentration of smoke particles entering the filter
v = linear velocity of the smoke
G, D and I = empirical constants for contributions from inertial
impaction, diffusion and direct interception, respectively.

where Dr = encapsulated pressure drop (mm H2O)

Q = volumetric flow rate (cm3/s)
L = filter length (mm)
S = fiber specific surface area (m2/g)

(unitless)
W = fiber weight (g)
C = filter circumference (mm)
r = polymer density (g/cm3)
d = fiber denier per filament (g/9 × 105 cm)
b = agglomeration factor (unitless)
F(a, q) = (2a - 1 - In a¢) + (4a - 3a2 - 2 - In a) Cos q

Cos q = TL/9 × 106W, where q is the average crimp

angle (°)
T = fiber total denier (g/9 × 105 cm)

Fig. 11.9
Single-fiber filter pressure drop model (Keith, 1978).
Figure 11.10 illustrates these particle capture processes (Keith, 1975).
Inertial impaction results when particles passing through a filter have
sufficient momentum to break from the flow path around a filament
and collide with it. Since inertial impaction is related to momentum,
filtration owing to this mechanism increases with particle velocity or
with particle mass. The relative contribution of inertial

Fig. 11.10
Smoke particle removal mechanisms (Keith, 1975).

 Page 367
impaction to cigarette filter efficiency is small (about 2% of total
removal) because particle momentum is low. Diffusional deposition
results when random particle motion causes collision with a filament.
Lower mass particles exhibit relatively more motion than larger
particles. Filtration resulting from diffusion is more prevalent at low
velocity because particle residence time inside the filter is increased,
allowing more chance for a particle to diffuse into a filament. About
65% of the total particulate removal is attributed to diffusional
deposition (Eaker, 1990). Direct interception results when particles
pass close enough to a filament to collide with it. This mechanism is
related to particle size and is independent of particle velocity. About
30 to 35% of the particulate removed by cigarette filters is attributed
to direct interception (Eaker, 1990).
From the considerations described above, removal efficiency is
expected to depend on both particle size (mass and diameter)
distribution and particle velocity. Figure 11.11 shows the theoretical
dependence of
 Fig. 11.11
Filter efficiency dependence on particle size (Keith & Derrick, 1965).

Filter Alpha Average pressure drop

type cellulose (cm H2O at 17.5 ml/s)
(% wt)
1 0 2.0
2 3.0 2.9
3 6.1 3.9
4 8.8 5.0
5 10.9 6.8
6 14.6 9.0

particle filtration on particle size (Keith & Derrick, 1965). The theory
predicts that removal efficiency is minimal at about the number
average particle size found in cigarette smoke. This means that
particles at the small and the large extremes of the size distribution
should be preferentially removed and that the aerosol exiting cigarette
filters should show a relatively higher concentration of particles in the
0.2 µ diameter range. Several researchers have attempted to
experimentally demonstrate particle size selectivity in cigarette smoke
filtration. However, some experimental results show no selectivity
based on particle size. The conflict between experimental results and
filtration theory may be due to method dependence of aerosol
measurements (Eaker, 1990).
The effect of smoke velocity on particle filtration is an important
consideration in vented-filter cigarette design. One consequence of
filter ventilation is reduced flow rate in the filter segment upstream
(adjacent to the tobacco rod) from the vent zone. As a result, the
smoke removal efficiency of that segment increases with ventilation
level due to increased retention from diffusional deposition.
Regardless of the ventilation level, the flow rate in the segment
downstream from the vents is constant. Thus, the efficiency of the
downstream segment does not vary with ventilation level. Since the
flow rate is higher in the downstream segment, its efficiency is lower
than that of the upstream segment. The total filter efficiency increases
with ventilation, but to a lesser extent than the increase found for the
upstream segment. Table 11.11 shows experimental results illustrating
the effect of ventilation on the efficiencies of each segment and on
that of the total filter (Lewis & Norman, 1986).
A secondary filter effect on smoke yields is selective filtration of some
of the semi-volatile constituents of
Table 11.11 Filter efficiency variation with
ventilation level (Lewis & Norman, 1986).
Filter Downstream Upstream
ventilation Total segment segment
(%) efficiency efficiency efficiency
(%) (%) (%)
0 55 30 36
10 56 31 37
20 56 28 39
30 56 28 38
40 57 28 40
50 60 29 43
60 62 28 47

 Page 368
smoke. Fiber filters, such as cellulose acetate, show preferential
retention of certain volatiles and semi-volatiles. For example, removal
efficiency for phenol is higher than that of the WTPM. Adding
triacetin to the filter increases phenol selectivity. Removal efficiency
for nicotine is lower than that for the WTPM. Volatile particulate
phase compounds retained to a lower extent than the WTPM are said
to 'elute' from the filter (Curran & Miller, 1969; Curran & Keifer,
1973). Selective filtration and elution are each aspects of preferential
retention. That is, constituents that have a high affinity for the filter
are retained while those having low affinity elute. Figure 11.12 shows
a depiction of this behavior (Curran & Keifer, 1973). Vapors from
semi-volatile compounds in smoke particles deposited on the filter
may be attracted either to the fiber or to smoke particles passing
through the filter. Compounds with higher affinity for the fiber will be
selectively filtered while compounds with higher affinity for the
aerosol particles will be eluted. Factors that influence preferential
retention include several properties of both the compound of interest
(evaporation rate, vapor pressure, solubility) and of the fiber. These
factors have been used to estimate selectivity (denoted

Fig. 11.12
Model for the elution process in filters (Curran & Keifer, 1973).
A semi-volatile (SV) compound which has been filtered can vaporize to
some extent from the fiber (A) or from an aerosol particle (B) and
condense either on another fiber (C) or on another aerosol particle (D).
 The SV compound that condenses on an aerosol particle vaporizes from
the particle (E), is removed by filtration as part of a filtered particle (F), or
escapes filtration as part of a nonfiltered particle (G). These processes may
occur a number of times before the SV compound escapes from the filter.
This overall process of a filtered SV compound escaping from a fiber by
entrapment in nonfiltered aerosol particles as they pass through the filter
has been designated 'elution'.
by Sx; selectivity is expressed as the filter retention of the compound
of interest relative to the particulate retention) for a range of semi-
volatiles shown in Table 11.12 (Morie, et al., 1975). Within a filter
fiber type (i.e. cellulose acetate or paper), preferential retention is not
greatly influenced by filter design adjustments. Affinities for some of
the semi-volatiles will vary with fiber type. As a result, filters of
similar WTPM efficiency made from different fiber types will yield
smokes of different composition and taste.
Because of elution, cellulose acetate filters have a slightly lower
efficiency for nicotine than for 'tar'. This difference is illustrated in
Table 11.13 for nonvented filters made with a range of tow items
(Parker & Montgomery, 1979). Measurements of the amounts of
WTPM and nicotine captured on filters and subsequently eluted have
been made (Curran & Keifer, 1973). These results allow calculation of
filtration efficiencies as if no elution occurred as well as the actual
efficiencies. Table 11.14 shows these calculations from the data of
Curran and Keifer (1973). After correcting for the measured elution,
efficiencies for WTPM and nicotine are the same. That is, elution
fully explains the differences in filtration efficiencies for nicotine and
'tar'. Table 11.14 also shows very little elution of WTPM (3.1% of the
amount captured on the filter). The bulk of the WTPM is not volatile
enough to have sufficient vapor for entrainment in smoke from
subsequent puffs. Thus, the net effect of elution on WTPM removal
efficiency is small.
Numerous additives and chemical treatments have been proposed for
selective filtration of cigarette smoke (Morie, 1977). However, dual
and triple element filters containing charcoal are the most common
examples in commercial use. Charcoal-containing filters are much
more efficient than cellulose acetate for removing compounds with
low (<152°C) boiling points (Morie & Baggett, 1975). Table 11.15
(Morie & Baggett, 1975) shows a comparison of cellulose acetate and
charcoal filter retention for several semi-volatiles. In addition, low
molecular weight aldehydes (formaldehyde, acetaldehyde, acrolein)
that are not affected by cellulose acetate can be reduced by 60 to 95%
(Keith, 1975). The type and amount of charcoal used influence
retention by the filter. Charcoal differs by its specific surface area and
pore volume. Retention increases with higher surface area and pore
volumes. Small amounts of high surface area charcoal (20 mg/filter)
can perform better than larger amounts (60 mg/filter) of low surface
area charcoal (Tokida, et al., 1985a). The method of inclusion within
the filter can influence retention. Charcoal can adsorb plasticizer used
in manufacture of

 Page 369
Table 11.12 Filter selectivity for smoke semi-volatiles (Morie, et al.,
1975).
Selectivity Vapor Solubility Vaporization rate
(Sx) pressure parameter (d) (Rv × 103)
(P)
1. Limonene 0.9 1.0 8.78 36
2. 0.9 36.0 8.34 946
Ethylcyclopentene
3. Eugenol 1.0 0.02 10.28 1.0
4. Methylindane 1.1 1.1 9.23 37
5. Cumene 1.2 4.6 8.60 146
6. 3-Heptyne 1.2 40 7.76 1052
7. Methylindene 1.3 0.5 9.48 16
8. Indene 1.3 1.7 10.07 68
9. 2- 1.5 11 9.72 350
Cyclopentanone
10. Ethylphenol 1.6 0.1 10.01 2.8
11. Pyridine 2.0 20 10.62 490
12. 2-Picoline 2.0 10 10.30 360
13. o-Cresol 2.2 0.4 12.50 12
14. Acetylfuran 2.2 1.1 11.64 56
15. 5- 2.5 0.5 11.52 24
Methylfurfural
16. Phenol 2.7 0.3 13.50 7.5
17. Furfural 2.8 1.6 12.19 47
18. Furfural 2.8 0.8 13.64 31
alcohol
19. Pyrrole 3.1 8.2 13.61 180

Table 11.13 Efficiency of non-vented

filters (Parker & Montgomery, 1979).
Filter tow ¢Tar¢ Nicotine
(dpf/TD × 10- efficiency efficiency
3) (%) (%)
1.6/50 53.0 49.3
1.8/42 49.6 43.9
2.5/48 49.6 44.4
5.0/40 34.7 27.8

Table 11.14 Calculated efficiency with and

without the elution effect (Curran & Keifer,
1973).
Filtration Elution as fraction
efficiency of
If no Capture on
Filter
elution (%) Actual filter Yield
tow
(%) (%) (%)
Nicotine:
5 dpf 30.2 25.8 14.6 5.9
1.6 dpf 55.6 43.1 18.0 17.6
WTPM:
1.6 dpf 54.8 53.2 3.1 3.6

dual cellulose acetate filters, thereby decreasing the effectiveness of

the charcoal. Filter ventilation enhances charcoal effectiveness
(Tokida, et al., 1985b). The enhancement is due to decreased smoke
flow and increased residence time in the upstream filter segment
(which contains the charcoal) and to decreased concentration of
materials being presented to the charcoal for adsorption.
c
Filter Ventilation
Filter (or 'tip') ventilation is provided by an air permeable zone around
the circumference of the filter. During puffing, part of the volume
enters through the filter vent region, effectively reducing the size of
the puff at the coal. Less tobacco is consumed with the reduced
volume puff. Accordingly, both gas and particulate yields are reduced
roughly in proportion to the degree of ventilation. Filter ventilation is
widely used in the industry as a means for reducing smoke yields to
levels that could not be reached with filters, permeable papers and
processed tobaccos alone. In the US, cigarettes with 'tar' yields of 15
mg or less now represent over 62% of the market (Maxwell, 1996).
Most of these cigarettes use vented filters.

 Page 370
Table 11.15 Selectivity of cellulose acetate and charcoal for smoke semi-
volatiles (Morie & Baggett, 1975).
Cellulose acetate + triacetin, Cellulose acetate +
PEG 600, glycerin Paper activated carbon
R (%) S R S R (%) S
(%)
Benzene 18 0.70 29 0.54 67 1.85
2,5-Dimethylfuran 39 0.93 50 0.76 75 2.44
Vinylcyclopentene 33 0.85 25 0.51 76 2.54
3-Heptyne 42 0.98 49 0.75 78 2.77
Ethylcyclopentene 33 0.85 38 0.61 75 2.44
Toluene 20 0.71 45 0.69 64 1.69
Paraldehyde 69 1.84 51 0.78 90 6.10
m- and p-Xylenes 11 0.64 43 0.67 52 1.27
Styrene + o- 15 0.67 35 0.58 49 1.20
xylene
Cumene 66 1.68 75 1.52 82 3.39
5-Methyl-2- 64 1.58 71 1.31 64 1.69
furaldehyde
Benzaldehyde 10 0.63 23 0.49 43 1.07
Benzonitrile 46 1.06 43 0.67 61 1.56
Limonene 19 0.70 53 0.81 35 0.94
Indene 87 4.38 63 1.03 88 5.06
p-Cresol 77 2.48 80 1.90 59 1.49
Methylindene 57 1.33 47 0.72 33 0.91
Naphthalene 59 1.39 45 1.11
R (%) is retention or filter efficiency for each compound.
S is selectivity.

Vented Filter Design

Air flow in vented filter cigarettes can be represented by the simple
electric circuit analogy shown in Fig. 11.13. The resistors in the
electrical analogy represent flow resistances of the filter, tobacco rod
and vent region. The vent flow is in parallel with the tobacco rod and
the filter segment 'upstream' (adjacent to the tobacco rod) from the
vents. From inspection of the circuit analogy, it is clear that the flow
(i.e. current) through the parallel paths must sum to the flow in the
filter segment 'downstream' (at the mouthend) from

Fig. 11.13
Electric circuit analogy for filter ventilation.
the vents. Increasing the resistance of the vents will cause decreased
flow in the vent path of the circuit and increased flow in the upstream
path comprising the tobacco rod and upstream filter segment
resistances. Since filter ventilation is defined as the fraction of the
total flow (i.e. the flow drawn at the mouthend) drawn through the
vents, increasing the resistance of the vent path decreases the degree
of filter ventilation. Conversely, increasing the resistance of the
upstream path increases the degree of ventilation. Designing a vented
filter entails modifying the vent path resistance by selecting an
appropriate means and component materials necessary for providing
the desired airpermeable region in the filter. Normally, the resistance
of the upstream path is not modified to adjust the ventilation level.
The upstream resistance must be considered in vented filter design
because it influences tipping and plugwrap specifications and because
upstream resistance variability affects ventilation variability.
The circuit analogy in Fig. 11.13 can be summarized as follows. The
total flow (Ft) at the cigarette mouthend is the sum of the flow
through the vents (Fv) and the upstream section (Fu):

 Page 371
Ventilation (D) is the percentage of the total flow that enters through
the vents:

The resistance of the vents (Rv) is a function of the pressure drop

across the vents (Pv). Vent path pressure drop is the sum of the
pressure drops across the individual components of the vented region
(i.e. the tipping perforations, plugwrap, axial filter component):

The resistance of the upstream section of the cigarette (Ru) is a

function of the upstream pressure drop (Pu). The upstream pressure
drop is the sum of the tobacco rod (Pr) and upstream filter segment
(Puf) pressure drops:

Using an empirical equation for the relationship between flow and

pressure drop

and the fact that the vent path pressure drop (Pv) at the vent flow (Fv)
equals the upstream pressure drop (Pu) at the upstream flow (Fu),
equations 11.7, 11.8 and 11.11 can be combined to show that dilution
is related to the upstream and vent path pressure drops measured at a
standard flow rate:

Note that equation 11.12 holds only for vent and upstream pressure
drops that vary linearly with flow. Despite this inaccuracy, equation
11.12 is useful as it easily demonstrates that ventilation level increases
when the upstream pressure drop increases or the vent path pressure
drop decreases.
In reality, nearly all vent pressure drops show a nonlinear response to
flow, The tobacco rod pressure drop, a component of the upstream
path, also varies with flow in a nonlinear fashion. Because of these
nonlinearities, accurate prediction of ventilation level is
mathematically somewhat more complicated than summarized above.
Further, a much more useful model for calculating ventilation level
results from considering the components of the vent and upstream
path pressure drops. Models that account for contributions of the
tipping, plugwrap and filter to the vent path pressure drop have been
reported (Durocher, 1984; Schneider, et al., 1984; Norman, 1985;
Dywer, et al., 1987). These can yield quite accurate estimates for
ventilation level, and since they account for effects of the various
component materials, such models can be used to develop materials
specifications.
Figure 11.14 shows a vented filter constructed with pre-perforated
tipping and porous plugwrap. In this example, the vent path pressure
drop is the sum of the pressure drops across the tipping, plugwrap and
axial section of filter beneath the vents. Tipping permeability is
directly proportional to the total area of the perforations. The pressure
drop across the tipping is related to the tipping permeability after
adjusting for the number of open perforations actually on the filter.
The plugwrap pressure drop is related to its permeability and the area
of the plugwrap beneath the vents through which the ventilating air
flows. This area is larger than the total area of the tipping perforations
but smaller than the area defined by the dry line (Durocher, 1984;
Norman, 1985; Dwyer, et al., 1987) (region where adhesive is not
applied to the tipping). Tipping with small diameter perforations will
give higher ventilation than a tipping of the same permeability made
with large diameter perforations. This is due to the larger effective
plugwrap area (and lower plugwrap pressure drop) that results from
flow through the smaller diameter tipping perforations. The axial filter
pressure drop can be calculated from normal filter pressure drop
measurements by adjusting them for the dimensions of the axial
section of filter beneath the plugwrap through which ventilating air
flows. This

Fig. 11.14
Vented filter components.

 Page 372
analysis enables calculation of the vent path pressure drop

from common component materials specifications, i.e

and empirically determined factors to account for the effective

plugwrap and axial filter areas and exponents (nt and np) for nonlinear
flow/pressure drop relationships of the tipping and plugwrap.
Calculation of flow rate as a function of vent path pressure drop
requires empirical determination of the flow/pressure drop
relationship for each component. Each can be expressed in the form

The upstream pressure drop is simply the tobacco rod pressure drop
plus the pressure drop of the filter segment upstream from the vents.
The filter component can be calculated from the vent location as
follows:

where L is the filter length and Lv is the distance from the filter
mouthend to the tipping perforations. The tobacco rod pressure drop
does not vary linearly with flow. However, the upstream filter pressure
drop/flow relationship is linear. Since in many cases the filter
component represents a much greater proportion of the upstream
pressure drop than the tobacco rod, nonlinearity associated with the
tobacco rod can be ignored. In this case, the upstream flow can be
expressed with equation 11.11.
To estimate ventilation level from this model formulation, equations
11.7, 11.8, 11.11 and 11.13 to 11.18 are used to find a numerical
solution (Norman, 1985). The model described is also useful for
analysis of finished cigarettes to assign causes for measured
performance that is out of specification or for determining component
adjustments needed to change ventilation level. Similar formulations
have been used for analysis of ventilation variability (Mathis, 1984;
Rasmussen, 1996).
It is apparent from Figure 11.13 that filter ventilation affects cigarette
pressure drop. For a vented cigarette, the pressure drop at the filter
mouthend is simply the pressure drop of the mouthend filter segment
(Pfm) plus the upstream path pressure drop (Pu). Note that the
upstream pressure drop arises from the upstream flow (Fu). If Pu is
measured at the standard flow rate, the cigarette pressure (Pc) drop
can be calculated if the ventilation level is known:

or

It is evident from equation 11.20 that the cigarette pressure drop

decreases with ventilation. At a constant ventilation level, cigarette
pressure drop can be increased by increasing pressure drop of the
tobacco rod or filter, by moving the vents toward the filter mouthend
or by using a dual filter design wherein the pressure drops of the
upstream and downstream segments can be independently adjusted.
Reduction of equation 11.20 yields:

where Phc is the cigarette pressure drop with the vents closed
(encapsulated). Rearranging equation 11.21 results in a form useful
for estimating ventilation level from cigarette, filter and tobacco rod
pressure drop measurements (Keith, 1979; Keith, 1980):

Although equation 11.22 was developed for estimating ventilation

level from pressure drop measurements, direct measurement
(Reynolds & Wheeler, 1977; Weatherly & Keifer, 1978) of vent flow
is more convenient and is now the common method for determining
ventilation. In the standard method (CORESTA Recommended
Method No 6, 1983), a constant flow (17.5 ml/s) is drawn at the
mouthend and a flow meter is used to measure the vent flow.
Since the flow dependence of vent path pressure drop is nonlinear,
ventilation level decreases when the flow rate drawn at the mouthend
increases (Lewis & Norman, 1986; Mathis, 1987). The degree of flow
dependence of ventilation is governed by the combination of tipping
and plugwrap used in the vent design. In general, on-line perforation,
where both the tipping and plugwrap are perforated, is the most flow
dependent design. Pre-perforated tippings with very small holes result
in the least flow dependence. Because many smoking machines
generate puff profiles in which flow varies, ventilation level measured
during a machine puff will show a minimum at the peak flow rate.
Average ventilation levels measured for unlit cigarettes with atypical
bell-shaped puff profile will be lower than those determined with a
steady flow. It is possible to design vent systems that Yield the same

 Page 373
ventilation level at constant flow, but different levels measured during
puffing (Lewis & Norman, 1986). Cigarette designers must account for
this behavior when reconfiguring products to make use of on-line
perforations and when changing method of off-line perforation.
Materials for Vent System Design
Processes for manufacturing vented filters can be classified into two
broad areas. The latest method is online laser perforation. In this case, a
laser system mounted on the cigarette maker tipper section perforates a
specified region of the filter in the finished product. Since the laser
perforates both the tipping and the plugwrap, nonporous plugwrap can
be used. The laser system can be adjusted for number of holes, and for
beam power and pulse width, which determine the hole size (Helms &
Lorenzen, 1984; Alliotti, 1985). Some systems can be used with closed-
loop control where the ventilation level is automatically adjusted to
maintain the design target. The other widely used method is pre-
perforated tipping combined with porous plugwrap. Pre-perforated
tipping and porous plugwrap of specific porosities must be selected to
yield the desired ventilation level in the final product. Applying pre-
perforated tipping requires specially machined glue rollers that do not
apply adhesive to the perforated area of the tipping patch. Plugwrap
papers with porosity ranging from about 1000 to 26 000 CORESTA
units (cm/min at 10 cm H2O) are available. Tipping can be perforated
mechanically, electrostatically and with lasers to yield a wide range of
porosities. Practically speaking, perforation methods that yield very
small holes (electrostatic, micromechanical, micro-laser) can be used
for ventilation levels up to about 25%. Higher ventilation levels require
larger holes achieved with mechanical or laser perforation. Features of
on-line and pre-perforated vent design are compared in Table 11.16.
Paper used for plugwrap is made from wood pulp and a small amount
of filler, usually calcium carbonate. Basis weight typically ranges from
about 20 to 30 g/m2, with lower porosity papers having higher basis
weight. Filler content is about 12%. Plugwrap bobbins carry 4000 to
7000 m of paper, yielding around 30 000 to 60 000 filter rods
(depending on rod length). High tensile strength is necessary to avoid
paper breaks during high speed filter manufacture. Tipping paper is also
made from wood pulp and calcium carbonate filler with basis weights
from about 30 to 40 g/m2 and filler content of about 15%. Normally,
tipping is printed with either a simulated cork pattern or white coating
and with a brand logo. It can be coated with nitrocellulose to minimize
the tipping sticking to smokers' lips.
Ventilation Effects on Cigarette Performance
With vented filter cigarettes, the puff volume drawn at the coal (Vc) is
reduced relative to the draw at the mouthend (Vm) in proportion to the
ventilation level (D) as follows:

Because the reduced puff volume at the coal consumes less tobacco,
smoke yields are reduced roughly in proportion to the degree of
ventilation. In general, yields of light gases (e.g. CO, CO2) are reduced
to a greater extent than predicted from the ventilation level. Particulate
yield (WTPM) reductions are very close to the degree of ventilation.
Yields of some semi-volatile constituents are reduced to a lesser extent
than indicated by the ventilation level (Norman, 1974).
Table 11.16 Comparison of vent systems with on-line perforated and pre-
perforated tipping.
Feature On-line perforation Pre-perforated
Ventilation Continuously adjustable from ca Appropriate combinations for
range 25% levels from ca 10%
Ventilation On-line Requires specification of new
adjustment tipping/plugwrap
Vent location Machine changeover required New tipping, glue roller
adjustment required
Ventilation Low, 1% absolute Medium 3-5% absolute
variability
Cost Capital expense for equipment Perforation upcharge plus cost
offset by lower material costs of permeable plugwrap

 Page 374
Since ventilation reduces yields of particulates and gaseous
constituents, it is an important addition to filtration in the cigarette
industry's efforts to reduce smoke yields. Although filters could be
designed to remove more particulates than the current commercial
range, pressure drops from higher efficiency filters would be
unacceptably high. Because filters do not reduce gas phase yields,
very high efficiency filters used without ventilation create particulate
phase yields out of proportion with gas phase yields. Gas phase yield
must be in 'balance' with the particulate yield for acceptable taste.
Using filtration in combination with ventilation, low tar cigarettes can
be designed with an acceptable pressure drop and proportional gas
phase/ particulate phase yields.
A range of cigarette designs with the same tar yield can be made by
adjusting the filter and ventilation level as shown in Table 11.17
(Arzonico & Norman, unpublished). In this example, tobacco rod
specifications (i.e. cigarette paper, tobacco blend and dimensions)
were held constant. Filters with six particulate removal efficiencies
were perforated on-line to ventilation levels needed to obtain the same
tar yield for each filter type. As evident from Table 11.17, lower
efficiency filters required higher ventilation levels. Cigarette pressure
drop decreased due to increasing ventilation levels used in
combination with lower efficiency (and concomitantly lower pressure
drop) filters (equation 11.21). The wide pressure drop range illustrates
one of the problems with designing acceptable vented filter cigarettes.
If pressure drop is too high, the effort required to draw smoke from
the cigarette might be unacceptable. On the other hand, if the pressure
drop is too low, smokers are frustrated by the sensation of too little
effort. Finding the optimum 'resistance' to draw is important in the
design of good tasting products (Leonard, 1976; Kassman, 1984).
Numerous patented vented filter designs have been developed to
optimize draw resistance or yield extra flavor at high ventilation. A
few such designs are summarized in Table 11.18.
Another effect of higher filter ventilation is increasing puff count, as
shown in Table 11.17. Because ventilation causes reduced puff
volume at the coal, the 'dynamic' burn rate (i.e. the length of tobacco
rod consumed during a puff) is reduced. The 'static' burn rate (i.e.
length burned during smolder) remains constant with ventilation level.
Thus, ventilation influences the amount of tobacco burned during
puffing relative to the amount burned during smolder. For cigarettes
designed to yield the same 'tar' at different ventilation levels, the
higher ventilation design will yield less 'tar' per puff. Reduced yield
per puff may compound the lower acceptability of high ventilation,
low pressure drop cigarettes noted above. To counteract the puff count
increase, very low yield products often include higher levels of
expanded tobacco. The expanded tobacco reduces yields and allows
use of lower ventilation levels.
Because ventilation results in reduced tobacco consumption during
puffing, it also reduces yields of gas phase smoke components. Since
filters do not affect the fixed gases, the ratio of gas phase yield to 'tar'
yield is reduced with increasing ventilation level. Table 11.17 shows
this effect for carbon monoxide (CO) yields. In fact, CO yield is
reduced in excess of the amount indicated by the ventilation level.
This is due to CO diffusing out of the tobacco rod as the smoke travels
through it. Increasing ventilation level decreases the smoke flow rate
through the rod. The decreased flow rates increase smoke residence
time within the rod and allow greater opportunity for gaseous
diffusion to occur
Table 11.17 Performance comparison of cigarettes made to equivalent ¢tar
¢ yield by filter and ventilation adjustment (Arzonico & Norman,
unpublished results).
Tow item Filter Cigarette ¢Tar¢ filter Puff
(dpf/TD × ventilation pressure drop efficiencya ¢Tar¢ CO count
103) (%) (mm H2O) (%) (mg/cig)(mg/cig)
2.1/48 0 213 71.2 7.6 12.8 7.0
2.7/48 24 166 65.5 7.3 10.1 7.2
2.7/46 31 139 61.7 7.1 8.9 7.3
2.9/41 38 113 58.7 7.6 8.1 7.4
3.3/39 42 93 53.4 8.1 7.9 7.6
3.9/35 53 64 47.3 8.5 6.7 7.9
NONE 75 13 7.2 2.1 8.4
a Filter efficiency at zero filter ventilation.

 Page 375
Table 11.18 Patented filters designed to optimize draw or flavor at high
ventilation.
Filter performance Design summary Patent
Higher pressure drop Sealed inner cylinder directs Berger, USP
filter for use with smoke through filter periphery 4508525 2 April
ventilation 1985
Higher pressure drop Smoke flow directed through Silberstein, USP
filter for use with small diameter orifices 4393885 19 July
ventilation 1983
Flavor adjustable by Rotating filter element with Kallianos, USP
the smoker smoke flow channels containing 4677995 7 July
different flavors 1987
More flavor from Unfiltered smoke flows through Norman, USP
vented filters central tube in filter 3860011 14
January 1975
Enhanced taste from Impervious disc at filter Hale, USP
low yield cigarettes mouthend diverts smoke flow 4331166 25 May
away from filter center 1982

(Owen & Reynolds, 1967; Morie, 1976; Baker & Crellin, 1977;
Muramatsu, et al., 1977; Durocher, et al., 1978; Townsend & Norman,
1982). Gaseous diffusion appears to be limited by rate of diffusion
through the tobacco bed and to a lesser extent by the paper
(Muramatsu, et al., 1977). Presumably, other low molecular weight
gases (CO2, HCN, etc.) that show yield reductions in excess of
ventilation levels also diffuse through the rod.
Filtration efficiency increases with ventilation level (Mathis, 1982;
Norman, et al., 1984; Lewis & Norman, 1986). For example, the data
in Table 11.11 show that tar removal efficiency increases from 55% at
zero ventilation to 62% at 60% ventilation. The increase is caused by
reduced flow rate in the filter segment upstream from the vents.
Particulate. removal in cellulose acetate filters is mainly due to
diffusional deposition (flow dependent) and direct interception
(independent of flow). In the upstream segment, diffusional deposition
increases because of the reduced flow rates. Since flow remains
constant in the segment downstream from the vents, filter efficiency
of that segment does not change with ventilation. Thus, the total filter
efficiency

increases with ventilation level.

Filter ventilation reduces yields of semi-volatile smoke constituents,
but to a lesser degree than tar is reduced. Semi-volatiles (including
nicotine) are compounds that are vapors at temperatures found inside
burning cigarettes but liquid or solid at ambient conditions. Lower
yield reductions for the semi-volatiles arise from effects of reduced
coal puff volumes on condensation and redistillation processes within
the tobacco rod. During a puff, air enters at the paper char line behind
the coal and is heated. When the heated air contacts the tobacco,
nicotine and other materials vaporize and rapidly condense
downstream from the coal to form an aerosol. Some of the aerosol
condenses on the tobacco. In ventilated cigarettes, relatively more of
the material condensed on the tobacco during previous puffs is
entrained in the smoke due to increased residence time in the tobacco
rod. Thus relatively more of the semi-volatile constituents are
included in the smoke. Because of this, yield reductions of
constituents such as nicotine are lower than expected from the
ventilation level. Although most of the under-proportional reduction
of semi-volatile yields is due to smoke formation differences, some of
this effect may be explained by increased elution from the filter.
Elution from filters is expected to be flow dependent. Thus, elution in
the filter segment upstream from the vents should increase with
ventilation level. Recent studies indicate that elution increases from
about 18% without ventilation to about 23% at 55% ventilation
(Muñoz, unpublished).
Tobacco Influences on Cigarette Performance
Blended cigarettes in the USA typically contain two or three types of
strip tobacco, processed tobacco (i.e. volume expanded and
reconstituted), humectants and flavorings. In other countries, blend
formulations include only one tobacco type according to regional taste
preferences. For example, in the UK and in Canada, cigarettes made
with only flue-cured tobacco are preferred. In these instances, flue-
cured tobaccos

 Page 376
from different quality grades and harvests are combined. Although the blend
formulation influences both the qualitative and quantitative smoke
chemistry, tobaccos are usually selected for taste and smoking quality.
Physical characteristics of the tobacco rod, including rod dimensions,
weight, tobacco shred size and amount of processed tobacco inclusion,
influence smoke yield and other aspects of cigarette performance. The
weight of tobacco used in the rod is an important consideration both
economically and with respect to finished product performance. Since
tobacco is the most expensive cigarette component, reducing weight while
maintaining consistent product quality is an important consideration.
Because smoke yields increase in proportion to the tobacco weight,
reducing tobacco weight is an important tool for overall smoke yield
reductions, particularly in the manufacture of low-tar cigarettes. Volume
expanded tobacco was introduced to reduce tobacco weight while
maintaining acceptable rod firmness and smoking quality.
Effects of Tobacco Type on Smoke Yield
Different types of tobaccos (e.g. burley, flue-cured or bright, Oriental,
Maryland, etc.) characteristically vary in smoke yield. In addition to
differences in smoke yield, the quantity of certain smoke constituents varies
with tobacco type. For example, Table 11.19 shows a comparison of smoke
constituent yields of cigarettes made with bright and with burley tobaccos
(Griest & Guerin, 1977). As can be seen from this comparison, per cigarette
smoke yield differences are typically larger than those found after
normalizing for tobacco weight. Because tobacco density characteristically
varies by type, cigarettes made at a constant firmness will vary in weight
depending on the tobacco type. Such cigarettes will show different burn
rates and puff counts due to weight differences and effects of tobacco
composition on burn rate. For these reasons, smoke yield data from
cigarettes made of a single tobacco type are difficult to translate to effects
on blended cigarette yields. Smoke yield also varies widely with tobacco
variety, stalk position (or quality grade) and region or other conditions
associated with cultivation of the tobacco. In fact, these variations can be
greater than those found for tobacco type (Griest & Guerin, 1977).
Although the tobacco blend is certainly part of the overall cigarette design,
blends are not typically formulated based on smoke yield of the different
tobacco grades or types. Rather, blend design is dictated by each
manufacturer's proprietary knowledge, chemical and physical
characterization of the tobacco quality, availability of tobaccos from various
regions and cost. Given the degree of inherent variability of tobacco by
grade, belt and crop year it is a challenge to maintain a high level of
consistency in finished cigarette quality and remarkable that reasonably
good consistency is achieved.
Table 11.19 Constituents of cigarette smoke from bright and burley tobaccos (Griest &
Guerin, 1977).
Quantity
Per cigarette Per gram of tobacco burned Per liter of smoke
Constituent Bright Burley Bright Burley Bright Burley
TPM (mg) 36.22 24.92 46.13 36.42 111.2 102.8
Water (mg) 3.41 3.27 4.35 4.78 10.53 13.48
Nicotine (mg) 1.66 1.56 2.11 2.27 5.13
¢Tar¢ (mg) 31.16 19.65 39.71 28.70 95.85 81.12
CO (mg) 18.67 17.88 23.81 26.22 59.27 78.59
CO2 (mg) 54.27 45.17 69.21 65.23 172.3 198.5
NOx (µg) 78.5 870.4 99.5 1269 252.2 3937
HCN (µg) 393.7 238.4 503.1 350.7 1186 1038
Acrolein (µg) 82.7 80.5 105.8 119.0 253.7 345.4
Acetaldehyde (µg) 797.3 730.7 1020 1079 2453 3166
Formaldehyde (µg) 26.8 25.1 34.1 36.8 79.2 111.3
Isoprene (µg) 750 335 960 495 2325 1450
TPM phenols (µg) 163.3 84.25 208.8 123.3 522.8 351.0


 Page 377
Two types of processed tobacco are widely used in the industry.
Reconstituted tobacco is made either by a slurry process or by a paper
making process from scraps of a size range unsuitable for cut filler.
Expanded tobacco is made by impregnating the tobacco cellular
structure with a high vapor pressure liquid, followed by heating to
drive off the liquid. Expansion of the internal tobacco structure results
from vaporization of the liquid. Reconstituted and expanded tobaccos
influence smoke yields and physical characteristics of finished
cigarettes. As might be expected, smoke yields vary to a degree within
each of these processes depending on the types of tobacco used as
starting material and the processing methods. In addition, both of
these processes are used to reduce manufacturing costs.
Reconstitution processes typically result in some loss of natural
constituents of the tobacco. These losses are greater with the paper
making process than with the slurry process because of differences in
how the materials are handled during sheet manufacture. However, the
chemical composition of sheet made by either process is different
from the starting tobacco. Cigarettes made exclusively from
reconstituted sheet show smoke yield differences relative to cigarettes
made from cut filler of the same tobacco type. Table 11.20 (Halter &
Ito, 1978) shows a comparison of smoke yields per gram of tobacco
for cigarettes made from each type of reconstituted sheet process and
with cut filler of the same tobacco used to make the sheets. The
reconstituted sheets showed higher static burn rates than cut filler
made from strip tobacco. The density of the paper process sheet was
lower and that of the slurry process sheet was higher than that of the
strip tobacco, resulting in the cigarette weight and pressure drop
differences shown in Table 11.20. In commercial cigarette designs,
reconstituted tobacco is included up to around 20% of the blend,
which results in slight reductions to per cigarette smoke yields.
The density of expanded tobacco is substantially reduced relative to
cut filler made from strip tobacco. As a result, cigarettes made
exclusively from expanded tobacco show considerably lower weight
and higher pressure drop. For example, Table 11.21 (Halter & Ito,
1978) shows a comparison of cigarettes
Table 11.20 Cigarette characteristics and smoke yields for cigarettes made
from strip tobacco and two types of reconstituted sheet (Halter & Ito, 1978).
Standard experimental Paper Slurry process
blend (SEB) process sheet sheet
Cigarette weight (avg) 1.226 1.150 1.399
(g)
Pressure drop (mm 96 79 54
H20)
Static burn rate 60.6 86.6 75.7
(mg/min)
Puffs/GTC 12.6 9.1 10.7
Peak combustion temp 831 834 829
(avg) (°C)
Particulate phase
Dry 'tar' (mg/GTC) 29.7 18.5 30.5
Alkaloids as nicotine 1.9 1.1 2.0
(mg/GTC)
Gas phase
Acetaldehyde 1.22 1.15 1.17
(mg/GTC)
Acrolein (µg/GTC) 124 125 87
Formaldehyde 40.9 56.8 40.5
(µg/GTC)
Hydrogen cyanide 229 121 190
(µg/GTC)
Nitrogen oxides 424 487 594
(µg/GTC)
Carbon monoxide 18.6 17.3 22.5
(ml/GTC)
Carbon dioxide 36.0 29.5 38.0
(ml/GTC)
 CO2/CO ratio 1.94 1.71 1.69
Condensate
Colorimetric phenols 6.55 5.44 6.60
(mg/g)
Benzo(a) pyrene 0.72 0.52 0.85
(µg/g)
Benz(a)anthracene 1.20 1.02 1.05
(µg/g)
GTC = gram tobacco consumed.

 Page 378
Table 11.21 Cigarette characteristics and smoke yields for cigarettes
made from strip tobacco and three types of expanded tobacco (Halter
& lto, 1978).
Standard RJR PM Freeze
experimental blend puffed expanded dried
(SEB ll)
Cigarette weight 1.160 0.804 0.768 0.874
(avg)
Pressure drop (mm 77.6 183.9 127.1 78.5
H20)
Static burning rate 0.068 0.063 0.057 0.081
(mg/min)*
Puffs/GTC 11.5 11.4 12.1 10.4
Peak combustion 821 806 822 816
temp (avg) (°C)
Particulate phase
Dry 'tar' (mg/GTC) 32.0 26.7 32.7 25.8
Nicotine 2.1 1.3 1.3 1.3
(mg/GTC)
Phenols (µg/GTC) 157 135 128 74
Gas phase
Acetaldehyde 1.16 1.39 1.28 1.50
(mg/GTC)
Acrolein 120 179 156 144
(µg/GTC)
Formaldehyde 37.0 35.3 38.7 52.5
(µg/GTC)
Hydrogen cyanide 418 339 523 371
(µg/GTC)
Nitrogen oxides 455 417 521 368
(µg/GTC)
Carbon monoxide 18.7 15.8 21.0 19.3
(ml/GTC)
Carbon dioxide 35.9 31.1 37.8 33.8
(ml/GTC)
 CO2/CO ratio 1.92 1.97 1.80 1.75
Isoprene (relative 1.19 1.26 1.28 1.25
values)
Condensate
Benzo(a)pyrene 0.71 0.76 0.45 0.57
(µg/g)
Benz(a)anthracene 0.93 0.88 0.65 0.94
(µg/g)
pH 5.15 5.09 6.26 4.87
* Values shown appear to be in units of g/min rather than mg/min as
indicated. Multiplication by 1000 will correct the typographical error
from the original source.
GTC = gram tobacco consumed.

made with tobacco from three expansion processes with those made
from cut filler of the same tobacco. Normalized for tobacco weight,
'tar' and nicotine yields from the expanded tobacco cigarettes are
somewhat lower than those for cigarettes made without expanded
tobacco for most smoke constituents. On a per cigarette basis, yields
from the expanded tobacco cigarettes are much lower. For example,
per cigarette 'tar' yields for the expanded tobacco cigarettes shown in
Table 11.21 are reduced by 32 to 42% and nicotine yields are reduced
by 53 to 59% relative to the control cigarette made from nonexpanded
cut filler. In commercial use, expanded tobacco inclusion generally
ranges up to about 50%. Because of the density reduction in expanded
tobacco, cigarette weights and per cigarette yields decrease in rough
proportion to the level of expanded tobacco inclusion.
Reconstituted Sheet Technology
Reconstituted sheet was originally applied in cigarette manufacture as
a means to reclaim tobacco scraps for use as cut filler. Subsequently, it
was developed as a blending tool to lower 'tar' yields and to optimize
taste. Reconstituted tobacco can also be treated to alter burn rates and
lower nitrate content. As a result of these features, the estimated
demand for reconstituted sheet is expected to grow at least 4 to 6%
per year to the year 2005 (Blackard, 1997). Browne (1981)
summarized the two processes used to make reconstituted tobacco
sheet. In the slurry or cast sheet process, ground tobacco is mixed with
a solution containing adhesive, combustion modifiers, mineral ash
modifiers, humectants and reinforcing fibers. The resulting slurry is
applied as a film to a moving stainless steel belt, dried

 Page 379
and either rolled onto bobbins or diced into strips suitable for making
cut filler. In the paper making process, water-soluble materials are
extracted from tobacco scraps in a closed vessel. The water extract is
concentrated, and humectants and combustion modifiers are added.
The extracted tobacco pulp is macerated and applied to a moving wire
screen to allow drainage of excess water. The water extract is applied
to the pulp and the sheet is dried, then either rolled onto bobbins or
diced for making cut filler.
The patent literature describes extensive development for both types
of reconstituted sheet process. As early as 1902 a process to make
paper primarily from stems and waste tobacco material was proposed
for manufacturing cigar wrapper (Marsden, 1902). This process
involved using steam in a closed vessel to separate the extractable and
cellular materials from the tobacco fiber, Pulp was made from the
extracted tobacco and glycerin was added to improve the consistency.
The pulp was made into paper and treated to make it resemble natural
leaf tobacco. Later, distilled water was proposed for use in the
extraction process to improve the extract (Moonelis, 1912).
A slurry or cast sheet method for making reconstituted tobacco was
proposed in the early 1940s (Sowa, et al., 1942). Samfield, et al.
(1954) proposed using mucilaginous plant gum to prevent tobacco
loss through fragmentation of the tobacco in the casting process. A
process identified as 'G-7' for commercial manufacture of paper-type
reconstituted sheet was developed in the early 1950s and later
improved (Thomasson, et al., 1988). Hungerford and Jurgensen
(1955) described detailed material preparation and methods for
forming cast sheets.
In 1956, Schweitzer proposed methodology for making reconstituted
sheets using technology similar to that used for paper making today
(Blackard, 1997), while another paper making process was proposed
by Wagner (1958). Parmele, et al. (1958) described a low moisture
process for reclaiming tobacco fragments or fines by grinding the
material and adding a solution containing binder and humectant
before passing the slurry through forming rollers. Improvements
proposed for a cast sheet system included addition of lithium chloride
or lithium bromide to the binder to increase flexibility and moistness
of the final tobacco sheet (Merritt, 1961). A technique for releasing
pectins from the tobacco was proposed to improve the binder
composition (Hind & Seligman, 1966). A summary of several
proposed developments from the late 1960s can be found in a patent
by Hind (1967a). Later, Gellatly and Uhl (1976) described a technique
for removing the potassium nitrate in a relatively pure state from
burley tobacco stems and proposed that the removed potassium nitrate
was suitable for use as a fertilizer, thus eliminating costly disposal
problems. Selke (1977) proposed a paper making process for
manufacturing reconstituted sheet from the whole tobacco plant. The
stalk and leaves were separated, refined by different processes,
combined and formed into a sheet by a conventional paper making
technique. A method was proposed for using tobacco dust in the
paper-type reconstituted sheet (Keritsis & Lowitz, 1981). Use of
ammonium salts of organic acids under high shear conditions was
proposed to destroy the alkaline earth metal cross-links with tobacco
pectin, making the pectin available to act as a binder in cast sheets
(Keritsis, et al., 1986). In an improvement proposed for the paper
making process, the aqueous extract was contacted with ammonia to
enhance the flavor of the reconstituted product (Thomasson, et al.,
1988). Prior to this process, reconstituted sheet had been treated with
ammonia after manufacture. Later, it was proposed that the aqueous
extract be contacted with diammonium hydrogen orthophosphate to
make a reconstituted sheet with improved flavor attributes (Young &
Bernasek, 1989). A similar improvement process for making a cast
sheet has also been proposed (Young & Fearrington, 1990).
In addition to the paper making and cast sheet processes, a method
and apparatus was described for making an extruded material that
could be incorporated into smoking articles (Tamol, et al., 1987). A
commercial process was developed for making an extruded smoking
material from a moist mixture of divided tobacco and a binding agent,
including locust bean and xanthan gums (Graves, et al., 1987). Two
other extrusion processes were later described (Luke 1988; Furuya, et
al., 1991).
Expanded Tobacco Technology
During the tobacco curing process, the leaf cellular structure loses
some of its original integrity and collapses primarily due to water loss.
For flue-cured tobacco, shrinkage is about 20% at the end of
yellowing, followed by another 52% during drying (Mohapatra
&Johnson 1972). Expanding tobacco increases its filling capacity by
partially restoring the original cellular structure. Filling capacity refers
to the tobacco's ability to occupy a finished cigarette rod with
adequate firmness, providing a sensation of thickness and substance to
the smoker (Durocher, 1996). The most

 Page 380
widely used process for expanding tobacco involves the use of carbon
dioxide to process blend components rather than the whole blend.
Over 150 patents have been issued related to the expansion process.
Expansion agents have included water, steam and various organic and
inorganic fluids. An early patent proposed impregnating tobacco with
a gas such as steam, air or carbon dioxide under pressure, followed by
release of the pressure to expand the tobacco volume by 15%
(Hawkins, 1929). Later it was suggested that tobacco stems could be
expanded by similar treatment of the whole leaf (Reed, 1939). Various
organic fluids have been suggested as suitable expansion agents,
including subjecting tobacco to organic solvent vapor followed by
drying (De Souza & Rice, 1962). Nitrogen has been proposed for use
as an expansion agent, where tobacco is pressurized with nitrogen gas,
then decompressed and heated with a gas to about 400°C (Ziehn,
1984). Other patents proposed impregnating moist tobacco with an
organic liquid such as a halogenated hydrocarbon, followed by steam
to expand the tobacco (Moore & Newton, 1971; Ashburn, 1972).
Because of environmental concerns, Freon is no longer used in the
United States as a tobacco expansion agent. As a consequence,
numerous agents have been investigated as potential replacements for
halogenated hydrocarbons, including propane, fluorinated
hydrocarbons, sulfur hexafluoride, pentane and mixtures of carbon
dioxide and propane (White & Conrad, 1982; Fagg, 1990; Kramer,
1991).
Numerous methods have been proposed for expanding tobacco
through moisture control and heat or steam treatment (Ergi, 1982;
Denier & Marshall 1986; Denier, et al., 1986; Hirsh, et al., 1988).
Many of the proposed methods involve rapidly heating the tobacco in
a hot gas stream. For example, one patent described treating tobacco
lamina with a high velocity, high temperature mixture of air and steam
to expand and stiffen the lamina (Keritsis & Sun, 1982). Moist
tobacco has been heated (Ashworth, et al., 1975), heated under
controlled conditions (Jewell, et al., 1977), dried under high humidity
(Mills, 1980), reordered (Hedge, 1984), conditioned (Wochnowski, et
al., 1982), moistened under storage (Ogawa, et al., 1991) and heated
in a gas stream so that the tobacco rises and falls in the gas stream
(Wochnowski & Thiele, 1978). Several patents have been issued
related to using steam to expand stems (de la Burde, 1965; de la Burde
& Harris, 1969; Buchanan & Madures, 1971; Psaras & Sachleben,
1978). Various methods for expanding tobacco using microwave
radiation have been proposed (Laszlo, 1973), and radiation of
moistened tobacco has also been proposed as an expansion process
(Neumann & Best, 1971; Stungis, et al., 1971).
Several patents described impregnating tobacco with agents that
chemically react to expand the tobacco or maintain the expanded
state, such as aqueous solutions of sugars, organic acids and inorganic
salts (Hind, 1967b). Impregnation of tobacco with cross-linking
agents followed by heating to expand and stiffen the tobacco has been
proposed (Keritsis, 1983). Attempts have been made to expand
tobacco using enzymes (Teng & Semp, 1979) and to improve the
degree of expansion using pectin releasing salts (Lendvay, 1978).
Treating tobacco with antioxidants prior to expansion to improve taste
characteristics has been investigated as well (Rudolph & Rodemeyer,
1982).
One of the first patents proposing carbon dioxide suggested
impregnating tobacco with ammonia and carbon dioxide followed by
heating the impregnated tobacco to expand it (Armstrong, et al.,
1970). Freeze drying processes for expanding tobacco were proposed
in the early 1970s (Smith, 1973). Later, the DIET (dry ice expanded
tobacco) process was developed where tobacco is impregnated with
liquid carbon dioxide (de la Burde & Aument, 1974; Sykes, et al.,
1979). The liquid carbon dioxide is converted to solid carbon dioxide
and the tobacco is then heated to expand it. Subsequent to the DIET
process, several patents were granted for various technologies related
to the use of carbon dioxide. These included proposals for dispersing
flavors in the expansion agent (de la Burde & Aument, 1979),
different technologies to introduce the agent (Rothchild, 1981) and
treating the impregnated tobaccos (Hibbitts, et al., 1978; Uematsu, et
al., 1991).
Tobacco Rod Physical Characteristics
Tobacco rod dimensions, weight, tobacco density and the shred size
influence rod firmness, pressure drop and smoke yields
(DeBardeleben, et al., 1978). Tobacco rod firmness is directly related
to the tobacco density. Increasing filling capacity, a measure of
tobacco specific volume under a pressure similar to that found in the
tobacco rod, will increase rod firmness at constant tobacco packing
density (i.e. tobacco weight/rod volume). Larger shreds, higher weight
and lower moisture will also increase firmness. However, lower
tobacco weights for reducing smoke yields require lower density
tobacco (higher filling capacity) to maintain acceptable firmness. The
primary means for reducing tobacco density is by volume expansion

 Page 381
through a process that increases the internal cellular volume of the
tobacco. Expanded tobacco density is reduced to about half that of
unprocessed tobacco. Blends of expanded and strip tobaccos achieve
significant filling capacity increases due to the decreased density of
the expanded material.
Tobacco rod pressure drop is a component of the finished cigarette
pressure drop (equation 11.21). As such, intermediate pressure drop
values are needed for consumer acceptance. That is, pressure drop
values that are too low or too high are unacceptable. For a given set of
dimensions, the tobacco rod pressure drop depends on the packing
density and the density of the tobacco (Schneider & Schluter, 1987):

where F is the flow rate of air drawn through the rod, A is the rod
cross-section area, K is an empirical factor determined by
manufacturing processes, rp is the packing density and rt is the
tobacco density. Thus, at constant packing density, reducing tobacco
density will increase the tobacco rod pressure drop. Maintaining
constant tobacco rod pressure drop with decreased tobacco rod density
requires reduced packing density so that the ratio rp/rt (i.e. the solid
fraction) remains constant. Changes to the tobacco rod pressure drop
influence levels of ventilation through the filter and through the
cigarette paper. Specifically, increased tobacco rod pressure drop will
increase ventilation through both routes. In addition, tobacco rod
variability is a major source of filter ventilation variance (Rasmussen,
1996). Thus, adequate control of the tobacco rod pressure drop is
important to consistent product quality.
Physical aspects of tobacco rod construction substantially influence
smoke yields. The most important determinant is the weight of
tobacco burned during puffing. Generally, increasing tobacco rod
weight will increase smoke yields. Thus, longer rods, larger diameter
rods or rods with higher tobacco packing density will yield more 'tar'
and nicotine. However, smoke yields are influenced by several factors
in addition to the amount of tobacco available. Static burn rates
decrease with increased tobacco packing density or with increased rod
diameter. Slower burn rates exaggerate the smoke yield increases
observed with increased tobacco weight because relatively more
tobacco is burned during the puffing cycle. The unburned portion of
the tobacco rod acts as a filter primarily through condensation of the
smoke as it is formed (Townsend, 1983). Relatively more filtration is
expected with longer rods or with higher packing density. Since
smoke velocity is reduced in larger diameter rods, more filtration is
expected as a result of increased diameter. As noted above, if rod
pressure drop increases with tobacco weight, ventilation levels will
increase. This factor would moderate smoke yield increases due to
tobacco weight increases. Thus, the effect on smoke yield of a
seemingly simple increase in tobacco rod weight is difficult to predict
because of the competing factors involved. That is, a weight increase
will cause more tobacco to be burned during puffing, but will also
cause more filtration by the tobacco and more ventilation, both of
which predict smoke yield decreases. Increased weight usually results
in increased smoke yields but the competing factors discussed above
certainly moderate and, in extreme instances, reverse the smoke yield
change observed.
Cigarette Design Challenges
During the past few years, issues other than those driven by consumer
demands have created cigarette design challenges. Such issues include
cigarette fire safety, environmentally degradable butts and
'secondhand smoke' or environmental tobacco smoke. The industry
has responded to each of these issues with research into feasibility of
products with the desired characteristics. Products that directly
address some of these issues have appeared on the market. For
example, low sidestream cigarettes such as RJR's Pianissimo and
Eclipse are currently on sale and the Rothman's brand, Passport, was
available in Canada in the early 1980s. The Philip Morris product,
Next, with very low nicotine yield was test marketed in the late 1980s
and early 1990s. The fact that some of these brands met with limited
success in the marketplace indicates that more work is required to
gain consumer acceptance and build in the performance characteristics
that address each issue. Since cigarette design components interact, it
is possible that improvements in one area of performance will reduce
effectiveness in another or will reduce some aspect of consumer
acceptance. A lesson learned over the years has been that reliance on a
single method of yield reduction (i.e. filtration or ventilation) is not
consistent with producing acceptable products. In fact, a systems
approach employing filtration, ventilation and processed tobaccos has
been used to reduce yields while maintaining smokers' acceptance.
Effective response to current issues will likely require new materials
and product designs and perhaps new technologies systematically
integrated to provide good smoking quality.

 Page 382
Reduced Ignition Propensity
In 1984 the USA Congress passed legislation that established a
technical study group (TSG) charged with determining the feasibility
of producing cigarettes that are less likely to cause fires when
carelessly dropped on upholstered furniture (Congress of the United
States, 1984). The TSG, composed of experts from various
government agencies, the furniture industry and the cigarette industry,
conducted studies at the National Bureau of Standards (now called
NIST the National Institute for Science and Technology). The TSG
found that small diameter cigarettes made with low tobacco packing
density (100% expanded tobacco) and low permeability paper were
less likely to ignite a range of laboratory fabric/padding mock-ups
(Gann, et al., 1988). Although the cigarettes described were
manufactured on a pilot scale, no assessment of commercial feasibility
was made, and issues regarding toxicological changes in the cigarettes
remained unresolved. Further, the test method used in the study was
not a standard test. That is, the test method was designed to show
differences in ignition propensity among the experimental cigarettes
but it did not indicate a realistic range of fabric or padding
performance. Under the 1990 Fire-Safe Cigarette Act (Congress of the
United States, 1990), NIST developed a method it claims will predict
the propensity of cigarettes to ignite upholstered furniture (Ohlemiller,
et al., 1993). However, it has been demonstrated that cigarettes with
low ignition propensity as measured by the NIST test can ignite some
fabric types. Also, cigarettes with 'high' ignition propensity by the
NIST test will not ignite other fabrics (Lewis, et al., 1995). In view of
these discrepancies, additional work is needed to adequately
understand characteristics of both fabrics and cigarettes so that a truly
predictive test method can be developed.
Environmental Degradability
Discarded cigarette butts can be viewed as a litter problem. The
materials used in manufacturing filter tips (paper, cellulose acetate)
degrade with lengthy environmental exposure. Recent efforts within
the cigarette industry have been aimed at developing a test method to
measure the degradation of filter materials (Brodof, 1996; Collazo, et
al., 1996). Developing faster degrading filter materials poses several
challenges. Ideally, new filter materials would have taste, smoke
removal and manufacturing properties similar to those of cellulose
acetate. It would be difficult to maintain taste characteristics with
radically different fiber materials because of smoke chemistry
differences caused by affinities of a new material for various smoke
constituents. It is almost mandatory that new materials be compatible
with existing manufacturing technology because of its level of
refinement and the industry's high investment. Research indicates that
design changes to cellulose acetate may improve degradability
(Wilson, 1996).
Environmental Tobacco Smoke
In 1994 the US Occupational Safety and Health Administration
(OSHA) reported highly controversial findings (US Federal Register,
1994) alleging health risks associated with exposure to environmental
tobacco smoke (ETS). Although a number of studies have shown that
potential exposure to ETS is very low in reasonably well ventilated
areas (for example, Jenkins, et al., 1996), publicity surrounding
OSHA's report created renewed interest in cigarettes with reduced
sidestream and 'second-hand' smoke. Sidestream smoke results
primarily from the tobacco burned during smolder. Cigarette designs
that yield appreciably less sidestream tar include special paper
formulations that interfere with particulate formation (as discussed in
a previous section) and can be made with lower tobacco weight for
further sidestream reduction. An example of such a design is the
Pianissimo brand currently sold in Japan. It shows a 50% reduction in
sidestream 'tar' relative to conventional products. Another low
sidestream product is the Eclipse brand currently in the test market in
the USA. Similarly designed products under various brand names are
available in other countries. The Eclipse design is shown in Fig.
11.15. In this design, mainstream smoke primarily results from
heating a bed of tobacco in thermal contact with a combustible heat
source. The

Fig. 11.15
Eclipse cigarette construction.

 Page 383
major constituent of the mainstream smoke particulate phase is
glycerin, which dissipates readily after exhaling. Since the burning
heat source has low tobacco content, the product generates almost no
sidestream smoke.
The cigarette design field has developed as new issues surface for the
industry. New materials will be needed to enable performance
characteristics that address these issues. At the same time, continuing
research of how cigarettes function and how the various components
interact will be required to understand the best way to integrate these
performance characteristics into acceptable products.
Acknowledgments
The author gratefully acknowledges Leslie S. Lewis for her insightful
discussions, skilful editing and assistance with locating reference
materials during preparation of this manuscript.
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Cigarette Filter Ventilation. FTR No 61. Tennessee Eastman
Company.
White, J.L. & Conrad, L.J. (1982) Process for increasing filling
capacity of tobacco. R.J. Reynolds Tobacco Company. Patent 4 531
529.
Wilson, S.A. (1996) Environmentally disintegratable cigarette filters.
Paper presented at 50th Tob. Chem. Res. Conf., Richmond, VA, 2023
October (abstract 20).
Wochnowski, W., Hohm, R., Liebe, R. & Muss, M. (1982) Apparatus
for expelling moisture from tobacco or the like. Hauni-Werke Korber
and Company. Patent 4 513 759.
Wochnowski, W. & Thiele, W. (1978) Method and apparatus for
increasing the volume of tobacco or the like. Hauni-Werke Korber and
Company. Patent 4 195 647.
Young, H.J. & Bernasek, E.B. (1989) Tobacco reconstitution process.
R.J. Reynolds Tobacco Company. Patent 4 987 906.

 Page 387
Young, H.J. & Fearrington, G.W., Jr (1990) Tobacco reconstitution
process. R.J. Reynolds Tobacco Company. Patent 5 099 864.
Ziehn, K-D. (1984) Process for improving the fillability of tobacco.
Reemtsma. Patent 4 577 646.

 Page 388

11C
Cigarette Production and Quality Assurance
J.L. McKenzie
McKenzie and Rains
Winston-Salem, North Carolina, USA

Chris Crawley
Fidus Instrument Corporation
Richmond, Virginia, USA
Introduction
This section of the monograph deals with the role of quality assurance
in cigarette production. The scope of the section is to describe the
purpose of quality assurance measurements, the importance and
source of specifications, measurement locations and frequency,
measurement equipment, and feedback and action programs. Also,
within this section, the term 'quality assurance' refers both to programs
and actions of the manufacturing operations personnel and to the
separate quality assurance or quality control department personnel. In
the terms used by some, these programs are respectively referred to as
process control and quality control.
Specifications
The basic function of quality assurance in manufacturing is to assure
that products are being made according to specifications. There are
two sources of specifications:
(1) design characteristics required to make the cigarette product
function properly and appear aesthetically pleasing, and
(2) production characteristics required to make the machining
properties of the product suitable.
Examples of the first group are tobacco weight, tobacco moisture, lack
of construction flaws such as filters which fall off, and package print
uniformity. Examples of the second group of characteristics include
such things as moisture of paper and board and the width of tear tape.
Of course, it is possible that a characteristic fits in both categories;
tobacco moisture and moisture uniformity are such characteristics.
Because specifications are the source for the determination of quality,
the beginning point for quality assurance in cigarette production is a
program for establishment of specifications that has the involvement
of production, engineering, marketing, marketing research, research
and development and purchasing department representatives. The
process of providing specifications should begin with marketing,
marketing research and research and development. These functions
provide the information required to design the product to meet the
requirements of the customer and consumer. Next comes the input of
the production and engineering functions who answer the questions
regarding the capabilities and requirement of the machinery. The
purchasing function translates the specifications of the manufacturer
into the specifications of vendors and provides critical feedback
regarding what are the capabilities of the vendor. Each company must
set up its own system based upon its particular situation. Describing
the exact procedures is not a proper objective of this monograph.
Rather, providing a checklist of some of the major specification
responsibilities of the functional areas will assist the reader in
establishing a quality assurance program or audit for an existing
program.
Specification responsibility
Function
Cigarette physical
Marketing
dimensions: length,
and
circumference or diameter,
marketing
tipping length, etc.
research
Cigarette performance Marketing
characteristics: particulate and
level category, menthol level, marketing
etc. research

 Page 389

Specification responsibility Function

Packaging physical Marketing
characteristics: packaging and
size, foil type, overwrap marketing
type. research
Graphic appearance: Marketing
cigarette print design and and
color, packaging design and marketing
color, foil appearance (bright research
or mat), tear tape design and
color, etc.
Cigarette physical Research
characteristics: blend and
composition, moisture level, development
draft resistance (tobacco rod
and filter rod), ventilation
level and ventilation design,
etc.
Materials' physical
Research
requirements: cigarette paper
and
porosity, filter wrap porosity,
development
packaging weight, size and
moisture, similar carton and
box dimensions and
requirements, etc.

Once these specification inputs have been provided by the marketing

and research functions, the manufacturing, engineering, and
purchasing functions must provide input regarding the feasibility of
the design. A checklist of responsibilities should include:

Specification responsibilityFunction
Machine capability and Manufacturing
capacity to produce and
product at the specified engineering
physical dimensions and
combinations.
Tolerance and control Manufacturing
capabilities of machinery. and
For example, what engineering
variances should be
expected in moisture level
or filter draft resistance.
Capability of vendors to Purchasing
supply the required (including leaf
tobacco and non-tobacco purchasing)
materials at the required
specifications.

For the particular details that should be covered within these checklist
areas, the reader is referred to the sections of this monograph covering
leaf physical properties (Chapter 9), tobacco blending (Chapter 11A)
and cigarette design and materials (Chapter 11B). Once all the inputs
to specifications are gathered, the specification document must be
written and published. Specifications must be numerically based and
as unambigous as possible. For example, a good cigarette weight
specification might be set on weight in grams of a 20 cigarette sample
taken at the mass flow and expressed as a weight average target and
acceptable range. Where the specification cannot be so easily
expressed numerically, as for example in the case of darkness of the
die print on a cigarette barrel, a good means of expressing
specifications would be to have light and dark color standards
available to the person making the quality checks and having a
tolerance limit of no cigarettes per sample of 20 outside the limit for
light or dark. More will be said on this subject in the next section
which covers measurement.
Measurement
To have a good quality assurance system, it is vital that the
measurement locations and sampling plans be appropriately chosen
and measurement reporting be understandable and action-oriented. It
is beyond the scope of this document to provide a thorough
explanation of sampling procedures and sampling plans. For that, the
reader is referred to sections 23, 24 and 25 of a handbook (Juran,
1988). Rather, a better use of this section is to point out to the reader
some of the particular technical matters to keep in mind when
sampling cigarettes. One commonly repeated error that quality
professionals must constantly be alert for is that specification limits
and sample estimates of a population mean are a function of sample
size. Often, those dealing with cigarette specifications and sample
means forget to change the limits appropriately when the sample sizes
are changed. The limits change proportionally to the inverse of the
square root of the sample size. The smaller the sample size, the poorer
the estimate of the mean and the wider must be the limits. Of course,
some reasonable sample size must be established based on the
economic impact of the sample size.
The first principle to keep in mind is that the sampling regarding a
particular characteristic should be accomplished as soon as possible in
the product life. For example, the first sampling for moisture control
should be taken at the maker and packer complex. Sampling in the
finished goods inventory leads to considerable delay in feedback to
the manufacturing personnel and is more difficult to trace and isolate
in terms of the production cut filler. Even worse, sampling moisture
from distribution locations introduces all sorts of complicating factors.
An opposite example is

 Page 390
measuring cigarette hardness. Measuring a cigarette fr esh off the
maker for hardness can result in inaccurate low hardness measures
because of the effect of temperature. Cigarettes should be held for 1
hour to allow for thorough temperature equilibration before measuring
for hardness. Another extreme is menthol content. If the menthol is
determined in the tobacco, the cigarette must be stored for up to 2
weeks to allow equilibration within the cigarette and the packaging
material. This measurement is further complicated in that menthol is
commonly applied to the tobacco, the filter or the foil. Equilibration
time and the important variable, menthol in the smoke, which is what
the consumer detects, are affected by the menthol application method.
The important physical parameters which must be measured on the
finished cigarette are:
Cigarette weight
Net tobacco weight
Moisture
Cigarette draft (ventilation open and ventilation closed)
Filter draft
Ventilation
Hardness or firmness
Tobacco rod circumference or diameter
Filter rod circumference or diameter
Sieve size
Filling capacity
Chemical and smoke measurements of importance are:
Menthol in cigarettes and smoke
Total particulate matter (required by regulation to be reported in some
countries)
Nicotine (required by regulation to be reported in some countries)
CO (required by regulation to be reported in some countries)
Cigarette puff count
Measurement of these characteristics, keeping these characteristics on
target and within control limits, will lead to a high probability that the
consumer will obtain the smoking characteristics the cigarette was
designed to provide. However, the principle remains that the final
product is a sum of its parts. The quality measurement program for the
cigarette must include some measurement of incoming tobacco and
non-tobacco materials. For the tobacco, it is important that the blend
be properly put together. The quality program must include process
control procedures to insure that the right tobacco blend ingredients
are used and that the scales and feed systems are properly maintained
to insure proper weight ratios and proper blending. Also, degradation
of strip and cut filler size must be held to a minimum. The non-
tobacco materials of particular importance, beyond the filter
characteristics mentioned in the checklist above, are cigarette and
filter plug wrap porosity and filter plasticizer content.
For a good explanation of the importance and interrelationships of
these parameters, the reader is referred to other sections of this
monograph and to the book The Design of Cigarettes (Browne, 1981).
A critical first line of defense in making good quality cigarettes is to
assure that the proper construction materials are present at the making
and packing complex. The quality program must include assignment
of responsibility that the proper wrapping and packaging materials are
at the complex and no others are on hand. In the bustle of activity
during production, one cannot take the chance that each bobbin, roll
or pallet of materials will be checked before it is put into production.
Also critical is the checking of product during change of brand
production. The program must include a thorough inspection of
materials during change over and inspection of product after the
change is complete. Nothing is more likely to make a customer
dissatisfied and suspicious of a company's quality program than to
buy a pack of cigarettes based on the label and to find a different
cigarette in the pack.
No quality program for cigarette production would be complete
without covering the visual and functional quality characteristics of
the cigarette.
The most important functional and visual characteristics of the
cigarette are those that make the product unusable by the consumer.
These characteristics must be measured in the cigarette making and
packing operation, and the most important ones are:
Things that make it impossible to light the cigarette or to keep it lit,
such as air pockets in the filter wrap, stem holes, broken and mashed
cigarettes, missing filters and paper splices.
Things that make the complete consumption of the cigarette
impossible or impractical such as filter falling off, fire cone fall out.
Things that make consumption of the cigarette unappealing, such as
stains or spots on the cigarette paper, foreign matter in the cigarette,
stains or dust on the filter, glue on the cigarette.
Things that make the packaging non-functional, such as missing tear
tape or pack glued shut.

 Page 391
Besides these characteristics of the cigarette, the quality assurance and
control program must strive for reasonable perfection. Characteristics
which make the final product more visually pleasing must be
included. These characteristics are always a matter of degree and are
rather subjective. How much can the color of the packaging vary, how
light can the print become and how out of line can the box become
before the product is unacceptable to the consumers? These are the
most difficult questions for the quality team to establish. The answers
are a function of the market and the price and quality category of the
product. Answering these questions properly are some of the most
serious items for a company to undertake. Placing the quality criteria
too high leads to unnecessary costs, and placing the criteria too low
leads to a high price in lower sales. Rather than be so presumptuous as
to give universal answers, the following checklist provides a
beginning point for internal company discussions:
Misalignment in cigarette and packaging materials
Wrinkles and rough cut in the cigarette and in the packaging
Missing or light print on the cigarette
Color variations in the packaging
Imperfections in printed material
Feedback and Action Programs
Feedback and action programs must be developed to insure that the
measurements taken by the operational personnel (control measures)
and those taken by quality assurance staff (audit measures) are defined
in terms that allow the source of non-conformance to be identified and
corrective actions implemented. Each company must have a clearly
written manual describing the inspection plan, reporting procedures,
action step responsibility and follow-up measurement procedures. The
establishment of responsibility for setting specifications and the well-
defined specification standards mentioned earlier are critical before
one can have a good action plan.
In particular, the reader is referred to standard quality assurance texts
such as mentioned before for reporting methods. Each operation
should have a quality council, or some other discussion forum, to
review quality issues and decide action plans. This council or forum
should include all departments responsible for establishing and
measuring product conformance to specifications.
Quality Assurance Instrumentation
This section is intended to give the reader information on the
development of the instrumentation business within the tobacco
industry. This is followed with information, by measurement
parameter, for the most common instruments used in the
manufacturing and quality assurance of cigarettes and filter rods. Most
consumers have little or no appreciation of the science and technology
within the tobacco industry. Most outsiders are astonished to learn the
level of applied technology in routine use within the industry, assuring
product quality, improving productivity and reducing costs. The
instrumentation area is no exception. Manufacturers are interested in
measurement of physical parameters using specialized instruments to
quantify the physical construction, consistency and product quality.
Today . . .
Since the early manufacture of cigarettes, there has been a need to
measure physical parameters (like circumference) as a direct and
secondary indication of product quality and consistency. Originally
the measurements required were minimal. Today this is not the case.
Cigarette making speeds in excess of 10 000 per minute and filter rod
production of 6000 per minute are routine in many of the world's
manufacturing plants. The physical measurements are of increasing
importance, especially as manufacturing sophistication and production
speeds increase. The trend today is to deploy instrumentation directly
in the manufacturing area and collect this data automatically for
immediate display to the machine operator, often in conjunction with
statistical process control (SPC) techniques. Sample sizes are
dropping to 5 to 10, from typically 20 to 30, but are being taken more
frequently. The time interval between samples is of increasing
importance in the implementation of SPC.
. . .and the future
The ideal way to ensure product quality would be to control all of the
constituent aspects of the manufacturing process on-line and measure
every unit produced. In many (most) cases this technology does not
exist, and some are impossible. Nevertheless, some on-line control
systems do exist. For example, basic weight control has been
successfully used for several years to control tobacco density and,
thereby, control tobacco rod weight. All high-speed cigarette making
machines are routinely supplied with on-line weight

 Page 392
control systems. Several systems have been made to produce on-line
circumference control. Originally these were pneumatic, which
worked rather well when the product was nonporous. As with all
pneumatic systems, a measurement error is introduced relating to the
porosity of the product to be measured. Efforts have been made to
produce on-line circumference systems using other technology. These
include infrared, CD cameras and lasers. Ventilation is routinely
measured on-line, and the trend is to employ on-line systems where
possible and use off-line instrumentation as a back-up. The demands
for instrumentation performance are becoming increasingly stringent.
Manufacturers are requiring laboratory standard measurements, not
only for the research and development and quality assurance
laboratories, but also for direct deployment in the manufacturing area.
This increased requirement for instrumentation in manufacturing
implicitly demands higher standards of instrument performance.
These are greater flexibility, higher reliability, minimal routine
maintenance and more automated operation. Special attention should
also be paid to automating calibration and easing work overhead for
routine tasks such as calibration verification.
The Market
If you visit tobacco companies throughout the world it is easy to see
the great diversity within the tobacco industry. Manufacturing is
incredibly diverse in size, scope and sophistication, making machine
production speeds run from 200 cigarettes a minute up to 14 000.
Factories vary from one maker per factory up to 100 high-speed
production machines. It logically stands to reason that the quality
assurance and quality control support functions also vary. This places
an understandably diverse requirement for different instrument
products for the same diverse market. While the trend is definitely
towards instruments of greater sophistication and automation, it is also
fair to say that there is, and will continue to be, a market for less
sophisticated manual instrumentation. A logical question is, then, why
automate? A good question, and the reasons are surprisingly diverse.
Certainly, reduced manning requires greater instrument automation in
conjunction with other manufacturing technology. It stands to reason
that as production speeds increase (and manning levels decrease),
there is a greater demand for instrumentation and information tools
directly in manufacturing. These can include automatic instruments
(feeding of samples and measurements) and automatic product
sampling (for cigarettes and filter rods). Information systems are also
increasingly deployed in manufacturing to acquire instrument data and
other production data. Often this information is now made available
on computer screen directly and in real-time to the production
machine operator. Often the operator is required to interpret this
information to control and improve product quality and consistency.
Successful implementation of SPC within production generally
requires a high degree of sophistication and integration of technology.
It is not enough to take samples and apply SPC rules for production
control. Production samples should be taken at regular time intervals
too. This ensures a more realistic picture of instantaneous samples
being statistically representative of the production population.
Increasingly, data integrity (and by extension information) for correct
analysis and action is critical today.
A Brief History
Early measurement tools, more gauges than instruments, were used to
measure diameter (circumference) and length. Later, liquid column
manometers for the measurement of circumference and pressure drop
were developed, based upon the pneumatic bridge principle. These are
low technology, simple instruments which are inherently reliable.
Many of these early instruments, built in the 1960s, are still to be seen
in facilities today. In the 1970s, the availability of affordable electrical
transducers and digital displays produced a plethora of manual digital
instruments to measure circumference, pressure drop and later
ventilation. Later in the decade, the application of microprocessors
produced the first truly electronic instruments. It is interesting to note
that almost all the early instrument development came from and was
driven from within the industry. Nearly all of the multi-national
cigarette companies (and some independent ones too) originally
supported R & D facilities well capable of designing and producing
instruments for their internal use.
Since the early 1980s, the development and application of
microprocessor technology has transformed instrumentation from a
workshop science into a specialist business led by a handful of
companies. Most of the cigarette companies have withdrawn from the
distraction of instrument development to concentrate on cigarette
manufacture. The applied technology is increasingly specialized,
requiring investment and overhead outside the more focused and
slimmer mission of today's tobacco industry. Today cigarette

 Page 393
companies prefer to purchase the instruments, systems and services
required and rely on the manufacturers for product support.
The microprocessor makes available instrument innovation and
development previously unrealizable. Instruments can be automated
(one of the most visible aspects), and in addition we see automatic
measurement, complex multiple calibrations, calculation and printing
of results (and statistics), hopper feeds and computer interfacing. One
of the greatest advantages is the ability to automatically measure and
adjust for shifts in tare, improving the instrument's calibration. During
this time low-'tar' cigarettes emerged in developed markets, prompting
the need to measure ventilation (or dilution). Several instruments
appeared around this time, often combining pressure drop and
ventilation measurements for cigarettes. By now cigarette
manufacturers are interested in routinely measuring several physical
parameters: weight, circumference, pressure drop and ventilation. For
further analytical purposes, hardness, length, moisture and paper
porosity are often added. In filter rod manufacture the key parameters
are circumference, pressure-drop and weight. These can be followed
in the laboratory by hardness, length, paper porosity and plasticizer
content. Ever increasing machine speeds, emphasis on product quality
and large scale manufacture stressed traditional quality departments.
The demand for data analysis, management information and reporting
pushed quality assurance departments to their limit and alternative
working practices were sought.
These demands passed rather quickly into the instrument market, and
it became clear tobacco companies wanted physical tests automated
and consolidated. Hence the emergence of test station like products.
These represented a large advancement in technology, combining
several tests within the same instrument on the same sample within a
few minutes. Individual samples were tested automatically. Results
and statistics were calculated and sent to a printer or computerized
data acquisition system. As with PCs, software emerged as the
controlling factor in instrument development. Today, much of an
instrument's personality (not to mention product development time)
and performance are determined more by software than hardware.
Today, quality control departments are invariably smaller as the
industry favors placing quality emphasis at the point of manufacture.
In practice this means the machine operator is invariably (or at least
increasingly) responsible for product quality. This has led to the
wholesale deployment of instrumentation and quality systems right in
the manufacturing department for the machine operators to use.
Specialized instruments with the flexibility to provide different
combinations of measurements with the capability to easily meet
changing requirements are becoming de rigeur in conjunction with
ever more sophisticated information systems.
Parameters Measured and Instrumentation
a
Weight
Cigarette weight is probably the most important parameter to measure
and generally the easiest. Tobacco is still the largest single cost
component of a cigarette and careful control of cigarette weight will
control tobacco consumption costs and allow analysis of tobacco
yields between processed batches. Manufacturers are also often
interested in measuring the cigarette's component parts for quality
audit and brand comparisons. During production both filter rod and
tobacco rod weight are routinely and continuously measured. Today
the electronic balance, with the capability for high accuracy and
precision, is the norm. Samples can be measured individually or more
usually in groups, frequently of 100 cigarettes, although these days the
sample size is trending downwards and so sub-samples of 50, 25 and
20 are seen. In cigarette production, group samples are frequently
taken, often by the machine operator, as a check of the cigarette
making machine performance. Today, most cigarette making machines
are fitted with an on-line basic weight controller which controls the
tobacco rod density. The off-line balance is used as back-up for the
on-line weight controller to monitor performance and to manually
adjust the on-line controller's set-point. In quality assurance
laboratories cigarettes and filter rods are often measured individually.
Sometimes they are classified into weight categories by weight sorting
instruments.
b
Circumference or Diameter Measurement (Also Called Size)
This is another crucial parameter for the cigarette industry
predominantly driven by material cost, but product consistency and
machine productivity are also important factors. Increasingly, related
parameters to laser circumference, like ovality and roundness, are
being requested. These relate more to peripheral cir-

 Page 394
cumference issues like cigarette end appearance and improving the
assembly process of the tobacco rod to the filter. Control of these
parameters permits improved roundness in the manufacturing process,
aiding machine runability. Prior to instrumentation the industry used
size gauges, but these are not used these days. Pneumatic
circumference gauges followed and these can still be seen, although
they are becoming less common. In years past they were the
predominant circumference instrument. Their decline results from the
increasing use of porous materials (for cigarettes and especially filter
rods) and a requirement for greater accuracy and automation. A
material's natural porosity causes measurement errors which can vary
even within the same material, making correlations to compensate for
measurement errors uncertain. Today measurement resolutions of 0.01
mm are routine and permit process control within ± 0.1 mm, although
wider tolerances are also used. Filtrona's tape gauge, which uses a
linear voltage differential transformer (LVDT) transducer, performs a
true circumference measurement by wrapping a mylar tape around the
product. Since 1994 laser gauges have outsold any other
circumference instrument. More and more customers are migrating to
laser. Besides unquestioned precision, they have the additional
advantage of improved calibration stability. In routine operation laser
gauges rarely need recalibrating. Laser gauges are the only
instruments capable of providing roundness and ovality
measurements.
c
Pressure Drop (Also Called Resistance-to-Draw (RTD) or Draft)
Pressure drop is a routine measurement parameter for cigarettes and
filter rods. For cigarettes, it is important to quantify the actual
cigarette pressure drop as perceived by the consumer, often referred to
as holes-open pressure drop, as the actual cigarette pressure drop is
reduced by the amount of ventilation, normally at the tip section.
Many manufacturers are also interested in holes-closed pressure drop,
as this is an indirect measurement of the filling power of the tobacco
rod and the filtration capability of the filter tip (if fitted). With filter
rods, pressure drop is measured as a manufacturing control parameter.
The filter rods are produced and measured as pencil-like lengths and
are sent in this form to the cigarette maker combiner where they are
cut into either fours or sixes as they are assembled into finished
cigarettes. Filter rod pressure drop is a primary parameter and part of
any cigarette design. Filter rods are, by their very nature, porous and,
if wrapped with a porous paper plugwrap or no wrap at all (as in the
case of nonwrapped acetate, NWA), then it is critical to measure this
parameter fully encapsulated. This is normally accomplished by
inserting the filter rod into a latex sleeve. In this way all of the air
entering the sample must exit at the other end without escaping. So
the latex sleeve fit is critical. While theoretically a relatively simple
measurement, pressure drop is often complicated by several factors:
The environment, as changes in temperature, atmospheric pressure
and relative humidity can have a significant impact on measurement
results.
The products to be measured are quite diverse and include a variety of
lengths, circumferences and porosities.
There have been many different instrument designs and developments
associated with the methods developed.
Varieties of sleeving materials are used to encapsulate the
measurement.
Early pressure drop testers were pressure-based pneumatic
manometers because, at that time, all regulators were designed to be
pressure based and vacuum regulators of sufficient stability did not
exist. Modifications to the basic design have been implemented over
the years to accommodate porous samples and a wider range of
circumferences. The CFO, or critical flow orifice, dominates as the
preferred method of pressure drop measurement. This is achieved by
creating a vacuum and accelerating the air flow through a calibrated
CFO to produce an air flow of 17.50 ml/s (or 1050 cc per minute) at
the instrument's measuring head where the product is measured.
Standardization of method and normalization of secondary calibration
standard methods are resulting in less variation between instruments
from diverse locations. This is particularly helpful to multi-national
companies producing the same products in different locations and
countries. Instrument manufacturers are now producing intelligent
instruments with auto-calibration features to automatically
compensate for variations caused by the operating environment. These
technological breakthroughs reduce instrument support overhead
requirements, particularly when instrumentation is deployed in
manufacturing.
d
Ventilation (Also Called Dilution)
This is of increasing relevance resulting from the introduction of
lower 'tar' or light cigarettes. In mature cigarette markets the
percentage of low 'tar' cigarettes

 Page 395
is often quite large and a growing segment of the market. Ventilation
is achieved by diluting the cigarette's total air flow. Usually this is
accomplished by perforating the tipping paper section of the cigarette.
Occasionally the tobacco rod (or envelope section as it is sometimes
known) also has some porosity, producing a measurable dilution of
the total air flow. The tip dilution causes a proportional reduction in
the holes-open pressure drop measurement, and this interdependence
between tip ventilation and holes-open pressure drop often means they
are both measured and studied together. Ventilation is normally
expressed as a percentage, no ventilation being a full flavor cigarette
and low 'tar' cigarettes having generally a high ventilation (although
this is not the only option for a cigarette designer). There are several
different ways to perforate tipping paper. The original mechanical
method has been superseded by electrostatic perforation and now laser
perforation, which can be done off-line, or on-line at the cigarette
maker. Ventilation is a relatively simple measurement, but one of the
least understood. One advantage of the use of a ratiometric
measurement technique is to make it largely independent of the
specified 17.5 ml airflow. Good ventilation measurements are
particularly sensitive to good sealing of the cigarette and small air
turbulence. Reliable results can only be obtained with off-line
instrumentation. An emerging trend is the introduction of automatic
calibration with obvious advantages.
e
Firmness (Also Called Hardness) Including Filling Power
Considered a controversial measurement within the industry, its
relevance is much debated as a primary control parameter of the
manufacturing process. Over the years there have been almost as
many hardness instruments as tobacco companies. Fortunately, only a
fraction of these are commercially available. Considered a destructive
test, comparison of results can only be reliably achieved statistically.
Units of measure also vary and include percentages, absolute
measured distance and comparative relative units. The product can be
measured in groups or individually. Individual measurement is
considered the most reliable method with the greatest integrity. Filter
rod firmness is an indirect measurement of its final suitability for use.
Normally a target range is set as part of the product's specification.
Reliable firmness measurements can only be achieved after the rod
has cured (meaning after the plasticizer has 'hardened') in the rod.
This is generally considered to be 24 hours after the rod was
manufactured. In some instances, filter rod resilience, which is a
product's ability to recover its shape, is also measured. Cigarette
hardness is an indirect measurement of the amount of tobacco in the
rod and how well packed or dense it is. This is often related to the
tobacco blend and the moisture content.
f
Paper Porosity (or Permeability)
The tobacco industry routinely measures the porosity of cigarette
paper, tipping paper principally at the zones of perforation and filter
rod plugwraps. These papers vary from very low (to almost
nonporous) values, as in the case of cigarette paper, to highly porous,
as in the case of ultra-porous plugwraps. Tipping papers lay in
between. There are many reasons to measure and evaluate the
porosities of papers used in the construction of cigarettes. They are a
critical design component which influence burn rates, subjective
quality indexes (like ash and coal appearance) and ventilation. The
emergence of light cigarettes has increased the attention and
importance applied to this parameter. There has been an increased
interest in measuring low porosity cigarette papers. The natural
variability of paper is well known, making it difficult to characterize
without several (often many) measurements. The measurement
technique is an evolution of the technology developed for ventilation
measurements. Great attention is applied to the engineering of the
measurement head and insuring incredible resolutions over a wide
range of product porosities. Advances in automatic pressure regulators
have enabled the introduction of faster and more automated
instrumentation.
g
Moisture (Including Water Content and Total Oven Volatiles)
The determination of moisture content in tobaccos, tobacco products,
cigarettes and filter rods is very important. Difficulties arise from the
exact definition of moisture; is it water only, or water plus other
liquids or volatiles? The traditional method for determining moisture
is the oven. Samples are routinely prepared and weighed before and
after placing the samples in a specific oven for a defined temperature
and time. Again this is not as simple as it may initially appear. There
are almost as many different ovens in use as there are tobacco
companies. Comparison of results becomes quite difficult. As with
many measurements, the predominant concern is precision in light of
the fact accuracy is questionable. Generally, as long as oven

 Page 396
methods are reproducible and sample preparation consistent, then
moisture (or more accurately total oven volatiles) can be determined
and compared with other samples using the same method (oven).
The advent of micro-processors has enabled the use of near infra red
(NIR) technology for moisture determination. The application of this
technology is now routinely used on-line in most tobacco processing
plants and cigarette primaries to measure moisture and provide a
means to control the process. The technology and its limitations are
generally well understood. Other methods include heating methods
(electric heating elements and microwave energy) in conjunction with
an electronic balance. This has the advantage of being much quicker
and less expensive, but invariably less accurate. One of the least
expensive methods is a capacitance method. Again, this is a low cost
method which is often less accurate.
In recent years, low energy microwave instrument technology has
been developed. The primary advantages are speed and accuracy with
results within 5 seconds to the same accuracy as the reference method.
The correlation between moisture and microwave energy absorption is
well understood and can be matched through calibration to any pre-
existing moisture determining method. At this time, the acceptance of
microwave technology has been limited by the relatively slow take up
of this new technology to unseat NIR technology and the lack of an
on-line system, although this is likely to change.
h
Length
Total cigarette and filter rod length is not routinely measured, but is
required to quantify brands and is part of a brand's specification. This
measurement is invariably done manually, although there are one or
two instruments which automatically measure length, even though the
product feed is invariably manual.
i
Measurement of Plasticizer
Plasticizer is the bonding agent used in the manufacture of cigarette
filters and is an important component to monitor during manufacture.
Too little, or no plasticizer, causes the filter rods to be too soft,
changing the product quality. Too much plasticizer causes the
cellulose acetate to partially dissolve, again irreversibly changing the
product's quality and rendering the filter useless. Other issues are
predominantly monetary. Plasticizer is relatively costly, particularly if
imported, and its correct application results in the cost saving and
improved product consistency. The ideal way is to measure plasticizer
on-line which is applied to the tow band, and although products have
been produced they have either been discontinued or are restricted to
metering (via a pump) the plasticizer entering the applicator booth.
Efforts to measure plasticizer on-line and off-line using NIR
techniques have resulted in mixed success. The most common method
in use today is the wet and dry method. This is accomplished by
turning off the plasticizer application, producing a small sample of
filter rods without plasticizer and comparing their weight with filter
rods from a normal production sample, using the same electronic
balance used for group weights. With this method, waste can be high,
and great care must be taken in preparing the dry sample.
j
Smoking Machines and Gas Phase Analyzers
Smoking machines, used by both industry and governments to
determine cigarette 'league-tables', are the important first step in
analyzing the smoking process and smoke constituents. While not
strictly speaking measuring instruments, they do adhere to
specifications of performance for the collection of total particulate
matter and mainstream gas phase and later laboratory analysis by gas
chromatography. The main gas measured within the smoking machine
is carbon monoxide (or CO) content. Other gas phase components can
be measured, although this is not routinely provided by the instrument
manufacturers. Smoking machines are available in single, 8 and 20
channel formats using both linear and rotary principles of operation.
Typically CO analyzers use the non-dispersive infra red principle of
measurement (NDIR) which has been well established over the years.
The principle is based upon the Luft cell, which uses an unweighted
bridge to produce an electrical output proportional to the amount of
CO. There are several gas analyzer companies commercially
producing such instrumentation. Care must be taken with calibration
to insure certified CO gases of known concentration are used.
k
Air Flow Measuring Instruments
The revised ISO 3308 specifications for smoking harmonization
require the air flows to be set up at the cigarette level. This air flow is
set to be 200 ml/minute ± 20 ml and therefore places a greater demand
for accuracy in air flow measuring instruments. There are

 Page 397
at least two instruments available capable of measuring such air flows.
Their range permits measurement of air flows in the exit ducting of
the fume hood as well. The measurement probes are fragile and the
instrument must be calibrated annually to comply with the revised
ISO 3308 procedure.
l
Cigarette Pack Seal Testers
How well a cigarette pack is sealed is an important consideration in
bringing a fresh product to the consumer. Measuring pack (and carton)
seal integrity insures an improved pack life from the time of
manufacture until sold to the consumer. Pack seal testing is used by
individual manufacturers to establish packing machine performance
and set up. It is also used in an audit capacity for brand comparisons.
Only in the last few years has effective instrumentation capable of
measuring pack seal integrity been commercially available. There are
manual and automatic instruments capable of manually measuring
individual packs or automatically measuring 10-pack samples and
reporting the data. There are at least two different instrument methods.
One method sets a known pre-defined pressure within the pack and
measures the flow leakage. An alternative method establishes a pre-
defined flow and measures the pack pressure. The measurements for
both methods are provided in engineering units and are evaluated
against 'good' and 'bad' specification limits.
Summary
It may be argued that instruments are not essential to cigarette
production, and this is true and adequately demonstrated by those
factories happily producing cigarettes with little or no
instrumentation.
As industry sophistication and maturity increases, instrumentation
certainly becomes considerably more of an essential requirement.
There are many contributing factors to this. Market requirements and
today's legislation force product evolution and brand proliferation.
There are competitive influences and the push to achieve superior
product quality. Product consistency and quantification, including the
competition, are of critical importance. Factory production volumes
are often high and beyond the scope of one individual to monitor.
There are market pushes to develop export business and pressures to
distribute product over enormous geographical regions. And all is
expected to arrive and be consumed with the same quality and
consistency as the day it was produced. Without instrumentation this
is impossible to verify and control.
High speed manufacturing operations, many of which implement SPC
systems, require instruments to maintain product consistency. Quality
these days is not measured in such clear cut terms as good and bad,
but more often as degrees of excellence.
References
Browne, C.L. (1981) The Design of Cigarettes. Hoechst Celanese
Corp., Charlotte, NC.
Juran, J.M. (1988) Juran's Quality Control Handbook; Section 23
Basic statistical methods, by Dudewicz, E.J.; Section 24 Statistical
process control, by Shainin, D. & Shainin, P.D. and Section 25
Acceptance sampling, by Schilling, E.G. & Sommers, D.J. McGraw-
Hill, New York.

 Page 398

Chapter 12
Smoke Chemistry
Richard R. Baker
British American Tobacco
Southampton, United Kingdom
Introduction
Tobacco smoke is an ever changing and extremely complex mixture
of chemicals. It is formed when tobacco, itself a complex mixture of
over 2000 chemical constituents, is burnt incompletely during the
smoking of cigarettes, cigars or pipes. Inside the burning cigarette or
other product during smoking, the tobacco is exposed to temperatures
ranging from ambient up to approximately 950°C in the presence of
varying concentrations of oxygen. Thousands of chemical substances
are generated, many of which arise from several distinct routes. The
components are distributed between the gas phase and particles which
constitute the smoke aerosol. The smoke itself is produced from the
smoking article in two distinct streams: mainstream and sidestream.
Mainstream smoke is drawn from the mouth or 'butt' end of the
cigarette when a smoker puffs on the cigarette. The smoke that rises
from the lit end of the cigarette, especially during the smolder period
between puffs, is known as sidestream smoke. Also included in
sidestream smoke are the constituents which diffuse through the
cigarette paper during the puff. Another form of tobacco smoke is
environmental tobacco smoke (ETS), which is smoke present in
ambient air as a result of smoking tobacco. It results from a
combination of sidestream smoke and exhaled mainstream smoke,
both greatly diluted by the ambient air.
In this chapter the formation of these three types of tobacco smoke
will be reviewed, together with their chemical and physical
characteristics. An enormous amount of work has been done on this
subject over about the last 40 years. Many experimental approaches
have been used in studying the various aspects of the subject. These
have included many detailed chemical analyses of smoke and smoke
from possible precursorloaded cigarettes, aerosol studies, pyrolysis
studies, investigations involving isotopically labeled chemicals, mass
transfer studies inside the cigarette and computer modeling of the
formation processes and their interaction. Since cigarettes rather than
cigars or pipes were used to generate the smoke in the majority of the
studies, the data in this chapter generally refer to cigarette smoke.
The subject matter in this chapter is divided into the following eleven
sections:
(1) The smoking of cigarettes
(2) Combustion processes
(3) Formation of mainstream smoke
(4) Formation of sidestream smoke
(5) Physical properties of tobacco smoke mainstream, sidestream and
ETS
(6) Fate of volatile tobacco constituents during smoking
(7) Chemical characteristics of tobacco smoke analytical
considerations
(8) 'Smoke pH'
(9) Smoke composition
(10) Generation of specific smoke constituents
(11) Environmental tobacco smoke
Section 1
The Smoking of Cigarettes
Cigarettes are, of course, smoked by humans and humans smoke
cigarettes in a variety of ways. It is well known that no two smokers
smoke in identical ways; no single smoker smokes in the same way on
all occasions (Thornton, 1978). The yield of components in cigarette
mainstream smoke depends upon the volume of puffs taken, the shape
of the puff 'profile' (the pressure/flow relationship whereby the
smoker may involuntarily control the volume of puff taken), the
number and frequency of puffs and butt length left at the end of
smoking (Creighton & Lewis, 1978; Darrall,

 Page 399
1988). The variations of smoking parameters observed with individual
human smokers are in the ranges of 20 to 80 cm3 for puff volume, 0.8
to 3.0 s for puff duration, 20 to 100 s for puff interval and 19 to 28
mm for butt length of unfiltered cigarettes (e.g. Creighton & Lewis,
1978; Schultz & Seehofer, 1978; Darrall, 1988).
In order to compare results between laboratories, smoke yields are
determined when the cigarette is smoked on a smoking machine with
a set of internationally agreed standard smoking conditions. The
general operational principle of all modern smoking machines is the
production of a puff of fixed volume using a motor-driven piston or
syringe. The agreed smoking conditions are a puff volume of 35 cm3
and 2.0 s duration, taken once per minute to a butt length of 23 mm
(nonfilter cigarette) or filter tipping paper plus 3 mm (for filter tipped
cigarette). This standard machine smoking procedure is adopted by,
inter alia, CORESTA, International Organization for Standardization
(ISO), Federal Trade Commission of the United States (FTC), and the
Laboratory of the Government Chemist in the United Kingdom. For
the purposes of this discussion, all of these procedures are identical.
The ISO and CORESTA standard methods are a very comprehensive
set of procedures which describe exactly how the cigarettes should be
sampled from the market place, environmentally conditioned prior to
machine smoking, the environmental conditions (temperature and air)
during the smoking, exact setting up and operation procedure for the
smoking machine as well as details of the chemical analyses
themselves (International Organization for Standardization, 1991af).
The development of the CORESTA methods, which were
subsequently adopted by ISO, has been reviewed by Thomsen (1992).
In the standard smoking method, the mainstream smoke exiting the
mouthend of the cigarette is immediately drawn through a Cambridge
filter pad (a glass fiber filter stabilized by an organic binder,
manufactured by the Cambridge Filter Corporation, Syracuse, New
York). This filter traps the aerosol particles present in the cigarette
smoke and is 99.9% efficient for particles larger than 0.1 µm in
diameter. The total particulate matter (TPM) in tobacco smoke,
sometimes called wet total particulate matter or cigarette smoke
condensate, is defined as that portion of the smoke collected on a
conventional Cambridge filter pad. 'Tar' is defined as the weight of
total particulate matter less the weight of nicotine and water (nicotine-
free dry particulate matter). The total nicotine alkaloids collected on
the pad are sometimes incorrectly referred to as nicotine yield,
although nicotine itself typically accounts for 85 to 90% of the total
alkaloids. Material passing through the Cambridge filter is
traditionally defined as the gas phase or vapor phase of cigarette
smoke. It should be noted that there is nothing absolute about the
separation of the vapor and particulate phases of the smoke aerosol by
the Cambridge pad. The materials collected on the pad are affected by
a host of factors including moisture content, temperature, flow
velocity and specific chemical interaction between aerosol
constituents and the fiber glass. The pad can collect some vapor phase
constituents.
Throughout this chapter, quoted smoke yields will have been
determined under these standard conditions. In addition, physical and
general measurements made on burning cigarettes will have invariably
used the standard smoking conditions.
The standard smoking methods are reliable, validated analytical
chemistry procedures for accurately determining cigarette mainstream
yields of 'tar' and nicotine. The first use of standardized machine
smoking procedures was made in the 1930s (Bradford, et al., 1936),
when cigarette manufacturers wanted to evaluate the mainstream
smoke yields of their products for quality assurance purposes, i.e. to
ensure product consistency. The development of smoking machines
and the associated analytical methodology since the 1930s have been
reviewed by DeBardeleben, et al. (1991). During the 1950s and 1960s
numerous groups advocated the measurement and publication of
cigarette mainstream smoke yields. This resulted in either voluntary or
legally required publication in many countries of the 'tar' and nicotine
yields of the cigarette brands available, obtained using the standard
smoking method. Machine smoking the same cigarette under
alternative smoking parameters may give, in absolute terms, larger or
smaller mainstream smoke yields. Furthermore, these different yields
may not vary in a linear fashion according to the parameter changed,
e.g. doubling the puff volume may not double the mainstream yield.
However, all currently available data indicate that product rankings
obtained under one set of smoking parameters are affected little by
choosing alternative smoking conditions provided that all the products
to be ranked have been smoked under the same conditions (Rickert, et
al., 1983; Darrall, 1988; Borgerding & Winkler, 1995).
Thus mainstream yields determined by the standard smoking methods
are appropriate for the ranking of cigarettes with respect to their yield.
The specified smoking parameters, i.e. puff volume, duration, interval,
etc., are well within established ranges of human smoking, as
indicated above. However, there has

 Page 400
sometimes been a misconceived public expectation that smoke yields
determined by the standard smoking methods should correspond to
the yields obtained from the cigarette by individual human smokers.
This is clearly unrealistic and, indeed, impossible. The machine
results are not necessarily predictive of the yields created by
individual consumers. Smokers not only smoke different brands
differently, but they also smoke the same brand differently depending
upon a host of human, social and environmental variables. In addition,
when the cigarette is smoked by machine with a constant puff volume
and duration, the yields of mainstream smoke constituents increase on
a puff-by-puff basis. This is due, inter alia, to the decreasing length of
tobacco rod as the cigarette is consumed and a corresponding decrease
in filtration by the tobacco rod. However, several studies have shown
that when humans smoke cigarettes they reduce their puff volume and
duration with each successive puff (Nemeth-Coslett & Griffiths, 1985;
Guyatt, et al., 1989; Kolonen, et al., 1992; Bentrovato, et al., 1995;
Reeves & Dixon, 1995). As Reeves and Dixon (1995) have pointed
out, human smokers are driven by a number of sensory cues, such as
smoke taste, irritation and impact, which are related to the yield of a
variety of smoke constituents in the puff. Smokers consequently
reduce their puff volume and duration as they consume the cigarette in
order to reduce the sensory effects of yields which would otherwise
increase with puff number.
A standard smoking method representing all smokers and conditions
is impossible. The situation is analogous to the methodology used to
measure fuel consumption for cars. These fuel consumption data are
determined under standard conditions of car velocity. No driver
expects to match these on every occasion when a particular car model
is driven; nor would two drivers expect to obtain identical figures
when driving the same car. What is achieved by fuel consumption
figures is a meaningful comparison of different models of car under
standard conditions.
In practice, with cigarette smoking, some smokers achieve 'tar' and
nicotine yields greater than those from the standard smoking methods,
while others achieve lower ones. Rankings based upon standard
smoking method results give the consumer good qualitative guidance
on relative cigarette mainstream smoke yields of dry particulate
matter and nicotine, but they are not necessarily predictive of absolute
'tar' and nicotine yields, under all circumstances, or for each
individual smoker. The rankings are, however, relevant in a general
sense to average human smoking. However, it should be remembered
that the smoking machine that generates the mainstream smoke does
not exhale, whereas the smoker who generates the mainstream smoke
exhales between 10 and 50% of the inhaled total particulate matter
(Dalhamn, et al., 1968).
There is a growing body of literature, based upon measurement of
nicotine and its metabolites in smokers' body fluids, that confirms on
average the broad ranking developed by machine smoking methods.
In other words, cigarettes with the highest nicotine yield from the
standard smoking method also produce the highest level of nicotine
and its metabolites received by smokers. Conversely, cigarettes with
the lowest nicotine yield from the standard smoking method produce
the lowest level of nicotine and its metabolites in smokers (Höfer, et
al., 1991; Rosa, et al., 1992; and comprehensively reviewed in Byrd,
et al., 1994).
Section 2
Combustion Processes
When the cigarette is lit, a burning zone is formed at the end of the
tobacco rod. Two distinct types of burning take place with the
cigarette, as well as with a cigar and pipe: puffing and natural smolder
between the puffs. During puffing, air is drawn into the cigarette
through the burning zone and mainstream smoke is formed. In the
interval between puffs a natural convection flow of air around the
burning zone in an upwards direction sustains burning and forms the
sidestream smoke. The approximate relationships of the major
combustion processes involved in smoke generation have been
summarized by Muramatsu (1981) and are illustrated in Fig. 12.1.
This simplified diagram applies to both puffing and smoldering
conditions. Tobacco is heated, causing moisture and volatile materials
to distil out of it, and its components to thermally decompose
(pyrolysis), both resulting in the generation of volatile gases and
leaving a residual, carbonized char. The char reacts readily with
oxygen in the air, via natural convection during smolder or forced air
flow during a puff. This exothermic combustion produces the simple
combustion gases carbon dioxide, carbon monoxide and water,
leaving the inorganic content of the tobacco as ash. The gases and
volatiles go on to form either mainstream smoke (during the puff) or
sidestream smoke (during smolder). Some of the energy generated
during the exothermic combustion feeds back to heat the unreacted
tobacco, resulting in a classic, self-sustaining combustion cycle.
Since temperature and heating rate of tobacco are so important when
considering smoke generation, some

 Fig. 12.1
Schematic diagram of major processes occurring in cigarette combustion (after Muramatsu, 19
temperature distributions inside the burning cigarette at various times during the
smoking cycle are illustrated in Fig. 12.2. These were obtained from a series of
experiments in which an infra red transmitting fiber optic probe or thermocouple w
inserted into different positions inside the burning zone to measure the temperatur
the solid and gas phases respectively (Baker, 1975a). The data in Fig. 12.2 refer to
the third puff in the smoking cycle with the cigarette smoked under the standard
conditions of a 35 cm3, 2 seconds puff taken once per minute. In the figure, the
position of the cigarette paper burn line is given the axial position of zero. Axial
distances in the unburnt tobacco rod are given as negative distances from the pape
burn line, while positions in the burning zone and ash are given as positive distanc
The general direction of gas flow through the cigarette is from right to left in the
diagrams.
The gas and solid phases are in near thermal equilibrium during the interpuff natu
smolder period, with highest temperatures of almost 800°C occurring in the center
the burning zone. During the puff the two phases have very different temperature
distributions near the surface, although they are similar in the central regions (cf F
12.2(b) and (c)). The highest solid-phase temperature (910°C) occurs at the burnin
zone periphery, about 0.2 to 1.0 mm in front of the paper burn line and 1 to 2 seco
after the start of the puff. This is where the air influx into the burning zone is grea
and, consequently, where the maximum amount of exothermic combustion is
occurring. The gas temperatures in the same region are relatively cool, varying
between 600°C and 770°C as the puff progresses. After the puff has ended, the so
phase
 Fig. 12.2
Variation of temperature (°C) distribution inside the
burning zone after the start of a 2-second puff
(Baker, 1975a). In (b), estimated air flow pattern
is shown. Thickness of arrow is proportional
to magnitude of air flow.
temperatures at the periphery of the burning zone cool from over 900°C to 600°C
1 second (cf Figs 12.2(b) and (d)). Within about 4 seconds the two phases have
attained quasi-equilibrium throughout the burning zone.
Based upon these temperature measurements and equivalent measurements in whi
the distribution of gases inside the burning zone was measured (Baker & Kilburn,
1973; Baker, 1981b), the general processes

 Page 402
illustrated in Fig. 12.1 are shown in the context of a burning cigarette
in Fig. 12.3. The processes occurring during cigarette combustion can
be rationalized as follows (Baker, 1981a; Baker, 1984; Baker, 1987a;
Baker & Robinson, 1990). In a cigarette, the rate of combustion of the
tobacco is controlled by the rate of mass transfer of oxygen to the
tobacco surface (Gugan, 1966; Baker, 1976a; Muramatsu, et al.,
1978). The viscosity and velocity of the gases in the burning zone
increase with temperature, so that the burning zone has a relatively
high draw resistance to gas flow (Baker, 1975b). Consequently, during
a puff, air tends to enter the cigarette at the base of the burning zone
near the paper burn line where the draw resistance is lowest. During
the puff, heat release at the base of the burning zone from exothermic
combustion of tobacco is greater than heat losses, and the peripheral
temperatures of the solid phase increase to well over 900°C.
Consequently, it is mainly the periphery of the burning zone that
advances during the puff (Egerton, et al., 1963; Jenkins, et al., 1977).
During about the first half of the puff, the pressure drop across the
burning zone increases rapidly, as the gas velocity, temperature and,
hence, viscosity rise (Baker, 1975b). Thus, as
 Fig. 12.3
The burning cigarette.
the puff progresses the volume of the cigarette consumed tends
towards a constant value, and an increasing proportion of the
mainstream smoke consists of air which has bypassed the burning
zone and entered via the paper (Egerton, et al., 1963). Since the
permeability of paper to air increases sharply as it starts to degrade at
about 300°C (Baker, 1976b), there is a large influx of air just behind
the paper burn line (Baker, 1976c). Therefore, since much of the
incoming air during the puff bypasses the central regions of the
burning zone, the gas and solid phase temperatures in the central
regions increase by a smaller amount than the temperatures at the
periphery (from just under 800°C to about 850°C (Baker, 1975a)).
When the puff ends, there is a greatly reduced transport of oxygen to
the surface of the burning zone and, therefore, a greatly reduced
amount of exothermic surface oxidation. The periphery of the burning
zone cools rapidly since it radiates heat to the surroundings; its main
source of heat is now the inner core of the coal. Oxygen diffuses into
the back of the burning zone via the cigarette paper (Mattina & Selke,
1977). The central region of the burning zone advances to re-establish
a relatively flat region at the back of the burning zone with relatively
constant temperatures across the diameter of the cigarette. Thus,
immediately after a puff there is often a time delay of up to 15
seconds before the paper burn line visibly moves.
During a puff, the volume of gases drawn out of the burning zone of
the cigarette is greater than the volume of air entering the burning
zone. This is due to the net formation of gaseous products. Various
measurements of this volume increase have been made, and a mean
value of 20% is a reasonable average estimate (Baker, 1981a).
Section 3
Formation of Mainstream Smoke
Within this general picture of the burning cigarette, the sequence of
processes that lead to product generation can be described. The
interior of the burning zone is oxygen-deficient and hydrogen-rich and
can be effectively divided into two regions (e.g. Hobbs, 1972; Baker,
1977; Baker 1981a) an exothermic combustion zone and an
endothermic pyrolysis/distillation zone. As air is drawn into the
cigarette during the puff, oxygen is consumed by combustion with
carbonized tobacco, and the simple combustion products carbon
monoxide, carbon dioxide and water are formed

 Page 403
together with the release of heat which sustains the whole burning
process. In this region, temperatures of between approximately 700°C
and 950°C are generated, and heating rates as high as 500 Ks-1 are
achieved (Baker, 1975a).
Immediately downstream of the combustion region is the
pyrolysis/distillation zone where the temperatures are between
approximately 200°C and 600°C and which is still low in oxygen
levels. Most smoke products are generated in this region by a variety
of mechanisms which are essentially endothermic (Baker, 1977).
Direct transfer from tobacco accounts for about one third of the
known smoke constituents (Green, 1977a). Examples of substances
that distil out of the tobacco include various saturated and unsaturated
aliphatic and terpenoid hydrocarbons, lactones, esters, carbonyl
compounds, alcohols, sterols, alkaloids, amino acids and aliphatic
amines and these have been reviewed extensively elsewhere
(Stedman, 1968; Green, 1977a; Hecht, et al., 1977; Schmeltz &
Hoffmann, 1977). Direct transfer depends on the volatility, functional
groups present and thermal stability of the compound involved
(Wakeham, 1972). Thus, menthol transfers efficiently to mainstream
smoke because of its volatility, and n-dotriacontane, a representative
tobacco aliphatic hydrocarbon, transfers efficiently because of its
thermal stability.
It has also been suggested that about a third of the transfer of i-amyl
benzoate is due to a nonthermal elution mechanism (Bingham, et al.,
1979). This is a mass transfer process where the concentration
gradient, and not heat, is the main driving force. Elution mechanisms
have also been proposed (Curran & Miller, 1969) to explain the
transfer of semi-volatile components from a filter. The component
vaporizes from a fiber and instead of recondensing on the fiber it is
trapped by an aerosol particle which subsequently escapes.
Surprisingly, some very nonvolatile and high molecular weight
substances also transfer to smoke, albeit very inefficiently, e.g.
inorganic salts, metals, sterols, carbohydrates and leaf pigments, as
well as some low molecular weight thermally unstable substances, e.g.
amino acids (Stedman, 1968). The presence of all these substances in
smoke implies that under thermal stress cellular eruption occurs and
rapidly ejects nonvolatile solids into the smoke stream before they can
thermally decompose. It is probable that such solid particles form the
nuclei onto which more volatile substances condense to form the
aerosol particles in smoke (Morie, 1977; Stöber, 1982).
The majority of the nonvolatile material is, however, unable to distil
out of the tobacco. This material, which includes most of the leaf
carbohydrates, sugars, polysaccharides (celluloses, starch, pectins),
polyphenols (lignin) and protein, is particularly prone to pyrolyric
degradation. These substances decompose to a variety of products,
including high-boiling compounds. Other classes of compound
formed from complex pyrolysis reactions include pyridines, indoles,
nitriles, aromatic amines, furans, phenols and carbonyl compounds
(Chortyk & Scholtzhauer, 1973; Jenkins, et al., 1975; Green, 1977a;
Hecht, et al., 1977; Brunnemann & Hoffmann, 1982; Baker, 1987b;
Schlotzhauer & Chortyk, 1987; Fenner, 1988; Jenkins, 1990). Of
course, many components in smoke can be formed by both direct
transfer and pyrolysis. Furthermore, carbohydrates and protein or their
degradation products can interact to produce many smoke components
(Green, 1977a). Since the whole of the pyrolysis/distillation region is
low in oxygen, only a relatively small amount of secondary oxidation
of products formed in this region can occur (Johnson, et al., 1975).
A highly concentrated, supersaturated vapor is generated in the
endothermic pyrolysis/distillation region of the burning zone, and
during a puff is drawn down the tobacco rod to form the mainstream
smoke. As the vapor is drawn out of the region it cools from about
600°C to almost ambient in a few milliseconds in the presence of
diluting air entering at the paper burn line. This brings the vapor of the
less volatile compounds quickly to their saturation point and
condensation follows immediately (Hobbs, 1972; Stöber, 1982).
Although some condensation directly onto the surface of the cooler
tobacco strands is likely to occur (Townsend, 1983), the condensation
will also occur instantly in the airborne state because of the presence
of a great number of condensation nuclei formed in the burning zone
(Morie, 1977; Stöber, 1982), as discussed above. Vapors of different
compounds may preferentially condense onto specific types of nuclei.
A dense aerosol consisting of growing droplet particles, which have
different composition and growth rates, is formed. Okada, et al.
(1968) have deduced that smoke particles are formed as the vapor
cools below about 350°C.
Townsend (1983) has presented a detailed model of the general
processes occurring during condensation of this smoke precursor
vapor leaving the burning zone. If nicotine is taken as an example, the
cooling vapor phase nicotine may either condense onto the newly
forming aerosol particles or condense onto the tobacco surface:

 Page 404

where N represents nicotine, v is the vapor phase, a is the aerosol

phase and d is that deposited on the tobacco rod. The aerosol particles
will then be filtered by the tobacco strands as the newly formed smoke
is drawn through the tobacco column. The nicotine deposited on the
tobacco strands can, in principle, revaporize and be trapped by an
aerosol particle (Curran & Miller, 1969). Townsend has described the
above scheme mathematically. The coefficients in the scheme (for
aerosol formation, condensation, filtration and elution) were found
iteratively which gave the best fit of the kinetic scheme to the
observed puff-by-puff data. It was found that the coefficient for
elution was zero, i.e. that an elution process is not necessary to fit the
observed experimental data. Furthermore, the coefficients for aerosol
formation and condensation of nicotine vapor are almost equal.
Figure 12.4 is a plot of the ratios of deposited vapor phase and
particulate phase nicotine to total nicotine (No) in the tobacco rod
behind the burning zone, obtained using the best fit coefficients. With
the exception of nicotine deposited on the tobacco, the vapor and
aerosol phase nicotine are theoretical plots. Nevertheless, the model
does illustrate very clearly vapor phase condensation and aerosol
formation in the few mm behind the cigarette burning zone.
Once the aerosol particles are formed, they will tend to follow the gas
flow through the tobacco rod. As they are drawn through the
decreasing temperature gradient at the back of the burning zone they
will grow due to coagulation and the continued condensation on their
surface (Creamer, 1979). However, owing to their physical size and
high concentration in the gas flow, the particles are simultaneously
subjected to thermal and mechanical effects which significantly
reduce their number. In the lower size range, droplets of sizes less
than about 0.1 µm have thermal diffusion rates which give a high
probability of collision with other particles or the tobacco surface. On
the other hand, particles with sizes near 1 µm have sufficient inertia
for them not to follow the tortuous gas flow through the tobacco rod
but to be removed by impaction onto the tobacco

Fig. 12.4
Ratios of deposited (Nd), aerosol
phase (Na) and vapor phase
(Nv) to total nicotine (No)
behind the burning zone
(Townsend, 1983).
surface (Morie, 1977; Keith, 1978).
In addition to these physical effects on the aerosol particles as the
smoke travels through the tobacco rod, a proportion of substances that
are permanently in the vapor phase will diffuse through the cigarette
paper and escape into the atmosphere as sidestream smoke (Baker,
1984).
Section 4
Formation of Sidestream Smoke
Some physical and chemical aspects of the sidestream plume in the
vicinity of the burning zone of a cigarette at the end of the interpuff
smolder period are illustrated in Fig. 12.5. The position of the carbon
monoxide plume was obtained in a series of experiments in which
small sampling probes, coupled directly to a mass spectrometer, were
placed around and inside the smoking cigarette (Baker, 1981b; Baker,
1982). The gas phase temperature distribution was obtained in
equivalent experiments using small thermocouples. The position of
the sidestream smoke plume was obtained by carefully photographing
the smoldering cigarette under controlled air flow conditions around
the cigarette. The gas phase temperature distribution outside the
cigarette in Fig. 12.5 is very similar to that obtained earlier by
Neurath, et al. (1966). The positions of the smoke and gas phase
plumes have also been confirmed with a Schlieren optical method
described by McRae and Jenkins (1992).
In the smolder period between the puffs, a natural convection flow of
air around the burning zone in an upwards direction (because of
buoyancy) sustains

Page 405
 Fig. 12.5
Concentration, temperature and velocities in
sidestream plume during smolder
(Baker, 1982; Robinson, 1987).
burning, but at much lower intensity than during the puff. Little
change occurs to the external temperature and oxygen distributions
when the puff is taken (Baker, 1982), indicating that the natural
convection stream around the burning zone and into the sidestream
plume is only slightly affected by the influx of air during the puff. The
combustion processes occurring on the surface of the burning zone in
the convection stream proceed independently of those inside.
The main products in the gas plume are carbon monoxide, carbon
dioxide, hydrogen and water the concentration distribution of carbon
monoxide outside the burning zone during smolder is shown in Fig.
12.5, and the forms of the profiles of the other gases and oxygen
depletion are similar. The carbon monoxide plume originates some 3
to 4 mm in front of the paper burn line, as do the external temperature
contours. The carbon monoxide concentration immediately above the
burning zone is higher than that just inside, and a similar situation
exists for carbon dioxide. Thus, sidestream carbon monoxide and
carbon dioxide are not formed only by the combustion products
diffusing out of the burning zone some must be formed on the
external surface from reactions with the oxygen convected around the
burning zone.
Figure 12.6 illustrates the variations during the smoking cycle of the
gas concentrations of oxygen, carbon dioxide, carbon monoxide and
hydrogen at a specific point in the sidestream gas plume. This point is
situated 1 mm above the surface of the burning zone and 3 mm in
front of the paper burn line. The plume hydrogen and carbon
monoxide concentrations at this point increase during the puff while
the carbon dioxide concentrations fall. At the end of the puff, the level
of all three products increases for about 1 second. This variation is
very similar to that found inside the center of the burning zone (Baker,
1981b), and is due to the outward diffusion of those products formed
inside the burning zone. The small rise in carbon monoxide and fall in
carbon dioxide during the puff are due, at least partly, to the
carbonaceous reduction of carbon dioxide

Fig. 12.6
Variation with time of sidestream gas concentration 1 mm vertically
above cigarette and 3 mm from paper burn line
(data from Baker, 1982).

 Page 406
to the monoxide which occurs as the temperature in the interior of the
burning zone increases as the puff progresses. When the puff ends, the
product formationtransmission balance inside the burning zone is
interrupted, resulting in a local build-up of gases in their formation
regions. These diffuse into the sidestream to deplete the local build-
up.
In contrast to the gas phase plume, the sidestream smoke plume
originates 0 to 4 mm behind the paper burn line, becoming visible at
temperatures below about 150°C (Fig. 12.5). This is the approximate
position of the tobacco pyrolysis and distillation region inside the
cigarette. Inside the cigarette in this region, a concentrated organic
vapor is formed. During the smolder period much of it will diffuse
radially out of the cigarette through the partially degraded cigarette
paper. As the vapor diffuses through the paper to the outside, it is
subjected to a sudden temperature decrease and dilution. These
conditions favor the formation of relatively small aerosol particles
compared to mainstream particles. As with gas phase sidestream,
there is a distinct increase in the emission of sidestream smoke
particles in the few seconds immediately following the end of the puff
(Dittman, et al., 1992).
Thus the conditions which generate sidestream smoke differ from
those producing mainstream in at least two important ways. Firstly,
the flow of air that generates sidestream is driven by natural
convection as illustrated in Fig. 12.3: both around the outside of the
burning zone (during smoldering and puffing) and into the back of the
burning zone through the cigarette paper (during smolder). This latter
air flow is in the opposite direction to the forced convective flow of
air during a puff. Secondly, the burning zone temperatures during
smolder, when the majority of sidestream smoke is formed, are lower
(reaching less than 800°C) than during the puff (reaching 900950°C).
Section 5
Physical Properties of Tobacco Smoke Mainstream, Sidestream and
ETS
The mainstream smoke emerging from the mouthend of the cigarette
during a puff is a highly concentrated, dynamic aerosol system. The
smoke particles are liquid, with water making up approximately 20%
of the droplet volume and, consequently, they have spherical shapes.
There are some 109 to 1010 particles per cm3 in fresh mainstream
smoke, making it an extremely dense aerosol. Initially, the particles
vary from less than 0.1 to 1.0 µm in diameter. Many studies have been
undertaken to determine the size distribution and number
concentration of the smoke aerosol using a variety of techniques, and
these studies have been comprehensively reviewed (e.g. Stöber, 1982;
Ingebrethsen, 1986a; McRae, 1990). Few techniques for measuring
size distributions will handle such a dense aerosol and many (but not
all) studies have greatly diluted the smoke prior to measurement. The
various techniques used have given a variety of results, with particle
number median diameters ranging from 0.16 to 0.60 µm and mass
median diameters ranging from 0.20 to 0.96 µm. While some of these
differences can be attributed to differences in the cigarette
characteristics and the measurement methodology used, a large part of
the discrepancy stems from the variation in time lapsed during
handling of the smoke prior to the size measurement (Keith, 1982).
The very high concentration of aerosol particles and their small size
are such that they will rapidly coagulate, resulting in sizeable
decreases in number concentration and increases in average particle
diameter within fractions of a second. In addition, in moist
environments the smoke particles can grow by absorption of water
vapor into the water-soluble materials in the particle. The particles can
also decrease in size by evaporation of volatile components, and this
is particularly the case when the smoke is diluted in air.
Two examples of the initial size distribution of particles in mainstream
smoke in the fourth puff of a plain cigarette are shown in Fig. 12.7.
Keith's data (1982) were obtained after applying a cyanoacrylate
fixation technique to the smoke emerging from the cigarette which
prevented subsequent coagulation of the particles. The 'fixed' smoke
particles were collected on a membrane filter and their sizes
determined by scanning electron microscopy. The data of Okada, et
al. (1977) were obtained using a light scattering technique after rapid
dilution. The distributions from these very different techniques are
similar and indicate a range of particle diameters largely between 0.05
and 0.5 µm with number mean diameters of 0.20 and 0.18 µm in the
two studies respectively. These distributions show a higher proportion
of smaller diameters than in many studies which have used aged
smoke and possibly inappropriate equipment. The distributions in Fig.
12.7 are near to the situation in fresh smoke where coagulation outside
the cigarette has not occurred to any significant extent.
The rate of coagulation of aerosols is second order in number
concentration (Fuchs, 1964) and is given by the following equation:

 Page 407

Fig. 12.7
Size distribution of aerosol particles
in fresh mainstream smoke.

where N is number of particles per cm3, t is time (s) and K is the

coagulation constant (cm3 s-l).
Integration of equation 12.1 gives the number of particles, N, as a
function of time and initial number per cm3 (N0):

The coagulation constant depends on the average particle diameter

and the spread of the size distribution for cigarette smoke an average
value of 6 x 10-10 cm3 s-1 is applicable (Stöber, 1982). Thus an initial
number concentration of 1 x 1010 particles cm-3 will decrease to
under half its initial value in 0.2 seconds and to less than 10% of its
initial value in under 2 seconds (see Fig. 12.8). This will be
accompanied by a corresponding increase in mean diameter of the
particles, and Ingebrethsen (1986a) has calculated the increase with
time using a theoretical treatment of Lee and Chen (1984) and an
initial particle size distribution typical of cigarette smoke. This
calculated size increase is illustrated in Fig. 12.9, along with some
data determined in three independent studies: the fixation/electron
microscopy technique of Keith (1982), Ingebrethsen's own data
obtained during a 2 second puff using a light scattering technique
(Ingebrethsen, 1986b), and a light

Fig. 12.8
Change in number of particles
in mainstream smoke
(N, cm-3) with time.

Fig. 12.9
Change in mean diameter of
mainstream smoke
 particles with time
(fourth puff).
extinction technique of Schneider, et al. (1988b). The theoretical and
the three experimentally determined curves are not identical and
would not be expected to be, since processes other than coagulation
will also be affecting the experimental results. The experimental curve
of Keith is similar in shape to the calculated curve. Keith varied his
times by passing the smoke immediately after the puff through various
lengths of tubing prior to analysis, and so should be measuring
coagulation effects only. Ingebrethsen (1986b) and Schneider, et al.
(1988a) sampled during the 2 second puff. Their experimental curves
are complex, initially showing size increases followed by size
decreases, possibly due to the complex combustion dynamics,
changing chemical composition and hence changing smoke refractive
index occurring in the early stages of the puff. However, all three
experimental curves do show the same general trends of a steady
increase in

 Page 408
particle diameter with time over at least the last second of the
experiment. This reflects the rapid effects of aerosol coagulation.
The size of smoke particles will also increase in moist environments
due to absorption of water vapor. This will be particularly important in
the respiratory tract where the relative humidity is estimated to be
99.5% (Ferron, 1977). The relative humidity of mainstream smoke is
60 to 70%. Although the experimental results obtained in different
studies to date are somewhat conflicting (Ingebrethsen, 1986a;
McRae, 1990), it appears that little particle growth occurs below 90%
relative humidity. Particle growth increases sharply with humidity
above 90% relative humidity, and calculations have indicated that
smoke particles double in size at 99.5% relative humidity (Ishizu, et
al., 1980). Generally, smoke particle growth is a complex process
dependent on the chemical composition of both the particles and the
surrounding gas phase. Schneider, et al. (1988b) have calculated that
substantial enrichment and depletion of particles by volatile
constituents, especially water, can occur in milliseconds. Both affect
particle growth.
The aerosol properties of the sidestream smoke plume have also been
extensively studied (e.g. Okada, et al., 1977; Ishizu, et al., 1980).
Where sidestream and mainstream characteristics have been measured
in the same study, the initial sidestream particles are found to be
smaller than the equivalent mainstream particles. For example, Okada,
et al. (1977), using a light scattering technique, reported sidestream
particles to have a geometric mean diameter of 0.11 µm with
logarithmic standard deviation 0.39, while the equivalent figures for
mainstream particles are 0.17 µm and 0.42. The generation rates of
the sidestream particles are reported to be in the range 1 to 6 × 109
particles s-1 (Ueno & Peters, 1986).
Both fresh mainstream and sidestream particles are lightly charged
electrically with about 30 to 50% of the particles being neutral and the
remainder being approximately equally divided between being
positively or negatively charged (Holmes, et al., 1959; Norman &
Keith, 1965; McRae, 1990). The proportion of charged particles
increases as the smoke ages over several minutes. It is surmized
(Holmes, et al., 1959) that while some of the electronic charge may be
produced in the burning process, the majority is acquired by collision
of the aerosol particles with ions. Of the charged particles, most carry
a single charge with only a very few having two or more charges.
Overall, the smoke is electrically neutral.
The sidestream plume accelerates as it rises above the cigarette. A
detailed determination of the velocity distribution of the sidestream
plume has been made using a laser Doppler velocimeter described by
Robinson (1987). In this technique, two laser beams are focused on a
given point in the plume and produce an interference pattern at their
point of intersection. Aerosol particles carried in the plume scatter
light from the interference pattern and characteristics of this light can
be used to calculate the velocity of the particles in the plume. The
results indicate that at a given distance above the cigarette there is a
distribution of velocity that obtained at 10 mm above the cigarette is
shown in Fig. 12.5(e), together with an estimate of the particle
concentrations in the gas and smoke plumes. The peak velocity of 410
mm s-1 occurs in the gaseous plume at 2 mm in front of the paper
burn line. The velocities in the smoke plume are generally less than
half this peak velocity.
Measurements at different distances above the cigarette show that the
plume is accelerating as it rises above the cigarette: the peak plume
velocity increases from 60 mm s-1 at 1 mm above the burning zone to
600 mm s-1 at 70 mm. This acceleration is accompanied by a falling
temperature of the plume with distance above the cigarette, which is
in turn accompanied by increasing density. This, along with the
observed acceleration, means that there is substantial increase in mass
flow rate in the plume with distance above the plume. This is brought
about by air being drawn radially into the buoyancy driven, natural
convection sidestream plume as it moves upwards. Detailed
calculations and mathematical modeling have confirmed that this
occurs (Robinson, 1987; Robinson, 1988).
As sidestream and exhaled mainstream smokes diffuse into the
atmosphere and away from the cigarette and smoker, they become
ETS. The originally concentrated sidestream and exhaled mainstream
smoke streams become greatly diluted; the sidestream smoke cools
and accelerates (Fig. 12.5), and various physical and chemical
changes occur in the smoke (Baker & Proctor, 1990; Eatough, et al.,
1990; Guerin, 1991; Guerin & Jenkins, 1992; Guerin, et al., 1992).
A variety of studies (Eatough, et al., 1986; Eudy, et al., 1986;
Hammond, et al., 1987; Benner, et al., 1989; Eatough, et al., 1989;
Ogden, et al., 1993) have shown that at least 95% of the nicotine in
ETS is in the vapor phase. Since nicotine in fresh sidestream smoke is
believed to be largely in the particulate phase (e.g. Proctor, et al.,
1988), it must evaporate rapidly out of the particles as the smoke ages
during initial dilution. Other semi-volatile organic substances such as
myosmine and pyridine also evaporate out of the smoke particles
during this dilution.

 Page 409
Studies by Black, Pritchard and co-workers (Black, et al., 1987;
Pritchard, et al., 1988) have shown directly that matter is evaporated
from fresh sidestream particles as they are diluted to form ETS. They
loaded 1iodohexadecane radiolabeled with 123I onto cigarettes. This
material has a boiling point of 380°C, typical of that of components
found in smoke particles. When fresh sidestream smoke was collected
from the smoldering, loaded cigarette, it was found that 5% of the
sidestream radioactivity was found in the vapor phase and 95% in the
particulate phase. On the other hand, when the cigarette smoldered in
a steel chamber of 14 m3 internal volume, 70% of the airborne
radioactivity was found to be in the vapor phase, and subsequent
radiochemical analysis indicated that there had been no chemical
degradation of the 1iodohexadecane in the environmental chamber.
Thus, the material had evaporated out of the sidestream particles
during dilution to form ETS.
Ingebrethsen, et al. (1986) and Ingebrethsen and Sears (1989) have
independently estimated that 20 to 30% of the original matter in
sidestream particles is lost by evaporation during the aging of ETS.
This estimate was calculated from measurements of the number and
sizes of sidestream particles diluted in a stirred 0.5 m3 stainless steel
chamber.
Benner, et al. (1989) have reported the evolution of changes in ETS
particle size distribution as the ETS ages over 4 hours. They generated
the ETS from the sidestream smoke of four smoldering cigarettes in a
sealed 30 m3 Teflon chamber. Some of their typical results are shown
in Figure 12.10. The particle size distributions at various times after
smoldering the cigarettes are all log normal. Immediately after the
cigarettes had smoldered, the ETS particles in the chamber had a
median number diameter of 0.11 µm (equivalent to a median mass
diameter of 0.26 µm). As the ETS aged over the next 4 hours this
median number diameter increased from 0.11 to 0.22 µm, while the
mass median diameter increased from 0.26 to 0.34 µm. These
increases are due to a combination of coagulation of particles and
removal of smaller particles by deposition onto surfaces of the
chamber. Surface deposition of ETS is the main route of removal in a
static environment and is a function of particle size, mixing rate, room
size and shape.
For their experimental conditions using a 30 m3 sealed chamber and
four smoldering cigarettes, Benner, et al. (1989) observed a total
decrease in ETS particulate mass of 0.5 mg m-3 over 3 hours. This
decrease was accompanied by a steady increase in total ETS gas phase
hydrocarbons (or, more precisely, total

Fig. 12.10
Changes in smoke properties as ETS ages (a)
 particulate number concentration;
(b) particulate and gas concentration
(max concentration indicated).
(Eatough, et al., 1987; Benner, et al., 1989).
gas phase substances measured using their flame ionization detector)
(Fig. 12.10(b)) equivalent to about 0.2 mg m-3 over 3 hours. This
increase must be due to the slow evaporation of hydrocarbons and
other volatiles out of the ETS particles, presumably from both the
airborne particles and those deposited on the chamber's surfaces. This
slow evaporation of hydrocarbons is in sharp contrast to the
evaporation of nicotine from the airborne particles, which occurs
almost instantaneously.
Section 6
Fate of Volatile Tobacco Constituents During Smoking
Before going on to discuss the chemical constituents in tobacco
smoke, this is a convenient stage to consider what proportion of
substances present in tobacco transfers to smoke. When tobacco is
heated during smoking, volatile tobacco components will distil out of
the tobacco and may also decompose thermally. These processes can
be readily studied by adding radiolabeled tobacco components
exogenously to the tobacco and following the fate of the radioactivity
as the cigarette is

 Page 410
subsequently smoked. Inherent in this technique is the assumption that
the radiolabeled material added to the tobacco surface (exogenous)
reacts to form the same products as does the naturally occurring
(endogenous) material. Jenkins (1990) has reviewed the available
evidence and concluded that, at least for the case of nicotine, the
assumption is justified.
Nicotine is probably the most thoroughly investigated tobacco
component in terms of its transfer to smoke. This is because nicotine
has such an important role in the smoker's sensory assessment of
mainstream smoke, in particular its 'impact' and 'irritation'. ('Impact' is
the localized sensation felt momentarily at the back of the throat;
'irritation' is a lingering, harsh sensory property of smoke). Both of
these parameters are involved in the smoker's perception of 'strength'
of the cigarette. The most comprehensive study on nicotine transfer to
smoke was published by Houseman (1973) in which radioactively
(carbon-14) labeled nicotine was added to a cigarette, the cigarette
smoked and the distribution of nicotine and radioactivity determined.
A summary of Houseman's main findings is given in Table 12.1,
indicating:
(1) 70% of the nicotine originally present in the tobacco burnt was
recovered intact after smoking (in mainstream and sidestream smoke
and on the butt).
(2) All the mainstream and sidestream nicotine was collected on a
Cambridge pad, which was taken as indicating that the nicotine was
wholly present in the particulate phase of the smoke.
(3) Twenty-seven percent of the original nicotine was converted to
other substances (by pyrolysis or oxidation reactions) which ended up
in the mainstream (0.5% in particulate phase, 4% in vapor phase),
sidestream (4% in particulate phase, 16% in vapor phase) and on the
butt (1.7%).
Other less comprehensive studies using radioactively labeled nicotine
(Larsen & Harlow, 1958; Jenkins, et al., 1971; Thornton & Massey,
1975; Jenkins, et al., 1977) have reported essentially similar findings
using cigarettes with different tobaccos or cigarette construction
characteristics (Table 12.2). The nicotine results from the five studies
quoted are reasonably similar, in spite of the fact that there is no
agreed means of collecting sidestream smoke and each group of
workers used a slightly different technique. In addition, Schmeltz, et
al. (1979) have shown that up to 11% of the nicotine in a burning
cigarette is converted by simple degradation to pyridines, about 3% to
other
Table 12.1 Fate of nicotine following smoking
of cigarette* (based on Houseman's
radioactively-labeled nicotine study
(Houseman, 1973).
Fate of nicotine Recovery
(%)**
Direct transfer to MS particulate 14.9
phase
Pyrolysis/oxidation products in MS 0.5
particulate phase
Direct transfer to MS vapor phase 0.0
Pyrolysis/oxidation products in MS 4.1
vapor phase
Direct transfer to SS particulate 37.0
phase
Pyrolysis/oxidation products in SS 4.1
particulate phase
Direct transfer to SS vapor phase 0.0
Pyrolysis/oxidation products in SS 16.3
vapor phase
Direct transfer to cigarette butt 18.5***
Pyrolysis/oxidation products on 1.7
cigarette butt
Nicotine and pyrolysis/oxidation 0.0
products in ash
Total 97.1
* 54 mm flue-cured tobacco column and 16
mm filter, smoked with 35 cm3, 2 s puff once
per minute to 21 mm butt length.
** Expressed as % of nicotine alkaloids
originally present in the tobacco burnt.
*** There will also, of course, be nicotine that
was originally present on the tobacco portion
of the butt.
MS = mainstream.
SS = sidestream.

organic volatiles and 12.5% is oxidized to carbon dioxide, all

generally being distributed predominantly to the sidestream. Schmeltz
and co-workers also compared these products obtained in a burning
cigarette situation to the products obtained from the thermal
degradation of radioactively labeled nicotine in nitrogen in a pyrolysis
tube at temperatures between 600 and 900°C. In contrast to the
burning cigarette situation, in addition to formation of pyridines,
under pyrolytic conditions the nicotine underwent much more
extensive degradation and re-arrangement to produce a variety of
quinolines, arylnitriles and aromatic hydrocarbons. Clearly, this
example illustrates that unless pyrolysis experiments are performed
under the dynamic conditions that are relevant to those that occur
during the smoking of a cigarette then their results can be highly
deceptive. In particular, the heating rate, sample size, atmosphere
(proportion of oxygen) and gas flow conditions, and residence times
of reactant and products in the pyrolysis furnace must be carefully
selected. It is not sufficient to use isothermal


 Page 411
Table 12.2 Distribution of radioactivity in smoke from carbon-14 labeled
tobacco distillable substances (based on tobacco consumed).
Mainstream Sidestream
Compound Particulate Vapor Particulate Vapor References
phase phase phase phase
Nicotine 20.1 5.3 53.4 21.2 Houseman (1973)
19.8 3.4 52.6 24.3 Larson & Harlow
(1958)
21.2 3.0 54.6 21.2 Jenkins, et al.
(1971)
23.5 2.2 52.6 21.7 Jenkins, et al.
(1977)
19.6 * 61.9 18.5 Thornton & Massey
(1975)
n- 27.0 0.4 60.6 11.9 Davis, et al. (1973)
Dotriacontane
24.7 13.5 48.3 13.5 Jenkins, et al.
(1970a)
30.5 0.9 55.3 13.3 Jenkins, et al. (1971
)
42.9 1.3 46.0 9.8 Jenkins, et al.
(1977)
Menthol 38.1 1.4 31.6 29.0 Jenkins, et al.
(1970b)
55.2 1.1 19.1 24.6 Eble (1987)
Glycerol 21.5 3.7 21.9 52.9 Larson & Harlow
(1958)
12.7 2.6 25.0 59.7 Best (1987)**
* Not reported.
** First three puffs only.

or low heating rate pyrolyses and assume that the results are
applicable directly to the conditions inside the cigarette.
Consequently, caution must be applied in interpreting the relevance of
pyrolysis experiments to processes that occur in a burning cigarette
situation.
Curran and Miller (1969) have studied the elution of nicotine
deposited on cellulose acetate filters in subsequent puffs. Cigarettes
loaded with radioactively labeled nicotine were smoked and the
nicotine filtered onto the cellulose acetate filters. The used filters
containing some smoke deposited labeled nicotine were then attached
to new nonradioactive tobacco rods, and upon smoking the fresh
smoke was found to elute the previously deposited nicotine.
Approximately 15 to 19% of the nicotine that had been deposited in
the filter was subsequently eluted by succeeding puffs. Elution is a
mechanism whereby the deposited nicotine vaporizes from the filter
fiber because of a concentration gradient, and it is trapped by an
aerosol particle and subsequently escapes from the filter.
The results of a number of studies in which other radio-labelled
components are added exogenously to tobacco are given in Table
12.2. Again, as in the study of Houseman (1973), the assumption is
made that the Cambridge filters used separate the vapor and
particulate phases of smoke, n-Dotriacontane, a C32 straight chain
alkane found in tobacco and other plants, has also been widely
studied. The work of Davis, et al. (1973) indicates that 97 to 98% of
the mainstream and sidestream particulate radioactivity is due to
direct transfer of the n-dotriacontane, the remaining 2% being due to
pyrolysis and oxidation products. The small amount of radioactivity in
the mainstream vapor phase and larger amount in the sidestream vapor
phase are due to pyrolysis and oxidation products. Jenkins, et al.
(1970a) have shown that these pyrolysis/oxidation products consist of
C1 to C10 hydrocarbons and oxides of carbon.
Results for menthol and glycerol (common flavor and humectant
additives to cigarettes) are shown in Table 12.2. The two quoted sets
of results for menthol differ in detail, but both indicate that the
mainstream radioactivity is found predominantly in the particulate
phase. Jenkins, et al. (1970b) have demonstrated that 99% of the
mainstream menthol is intact and the remaining 1% is pyrolysed to
menthane, menthene or oxidized to carbon oxides.
For all the substances quoted in Table 12.2, the mainstream
vapor:particulate phase radioactivity ratio is relatively small,
indicating that the substances transfer to mainstream predominantly
unchanged. In contrast, in the sidestream smoke the proportion of
vapor phase radioactivity is much higher, reflecting the higher
proportion of pyrolysis oxidation products in sidestream smoke.
(However, these results may partly reflect differences in collecting on
a Cambridge pad under mainstream and sidestream collection
conditions.)

 Page 412
Section 7
Chemical Characteristics of Tobacco Smoke Analytical
Considerations
Mainstream, sidestream and environmental tobacco smoke
components are distributed between the particulate phase and the
vapor (or gas) phase of the smoke aerosol. The ideal way of analyzing
tobacco smoke would be to unambiguously separate these two phases
and to analyze them immediately. Unfortunately, there is no
completely unambiguous way of separating the particulate and vapor
phases. Furthermore, some components are partitioned between the
two phases, and this partition changes with time, temperature and
dilution of the smoke. For the purposes of a generally accepted
definition, the portion of smoke which passes through a Cambridge
filter pad at room temperature is called the vapor phase and that which
is retained on the filter is called the particulate phase. However, the
trapping efficiency of a Cambridge filter is a function of a number of
factors, including the nature and amount of material being collected,
the flow through the filter and the temperature and moisture level of
the filter (Grob, 1965) as well as the condition of the cigarette at the
time of smoking (moisture content, etc.). For those substances which
are distributed between the particulate and vapor phase of smoke, only
partial trapping occurs on the Cambridge filter a few examples, taken
mostly from Sakuma, et al. (1978), are given in Table 12.3 for
mainstream smoke. In general, substances with a molecular weight
below about 60 tend to be predominantly in the vapor phase and
substances with a molecular weight above 200 tend to be wholly in
the particulate phase. The proportion of substance in the vapor phase
increases as the vapor pressure of the pure
Table 12.3 Distribution of some
mainstream smoke components
between vapor and particulate phases.
Component % in Particulate
phase
Acetaldehyde 2
Acrolein 7
Propionaldehyde 15
Benzene 22
Formaldehyde 32
m- 41
Dimethylbenzene
HCN 50
Acetone 66
Limonene 69
Nicotine 100

compound increases (Vekris & Hook, 1966). This distribution does

also depend on the polarity of the substance. For example, 41% of m-
dimethylbenzene (molecular weight 106) is in the vapor phase
whereas 100% of the more polar cresols (of similar molecular weight,
108) is in the particulate phase.
Other techniques for collecting the particulate phase of tobacco smoke
have been used and include electrostatic precipitation and jet
impaction, and the use of solid adsorbents such as activated charcoal,
silica gel, molecular sieves and Tenax. Cold traps and solvent traps
have been used for collecting the vapor phase of smoke (Dube &
Green, 1982). Each of these methods gives slightly different levels of
substances in the particulate and vapor phases of the smoke. Thus the
specific quoted division of substances between the two phases is
somewhat arbitrary and is dependent on the exact collection
methodology used.
In addition to these two principal phases of smoke, reference is often
made to the so-called semi-volatile fraction. This term was first used
by Williamson, et al. (1965) to denote materials which are retained by
the Cambridge filter at room temperature but can be volatilized from
the filter at a defined temperature without appreciable decomposition.
This temperature has tended to differ from study to study, but is
typically between 100 and 200°C, and roughly 5% of the total
material on the Cambridge filter is volatilized (e.g. Grob, 1965;
Williamson & Allman, 1965; Grob & Völmin, 1969; Grob & Völlmin,
1970; Neurath, et al., 1971; Neurath, 1972; Grob, 1973; Baggett, et
al., 1974; Mauldin, 1976; Norman, 1977). The semi-volatile fraction
is, therefore, not precisely defined and differs slightly in different
studies, but in general it consists of about 300 smoke components
having boiling points in the range 70 to 300°C, such as hydrocarbons,
alkylbenzenes, naphthalenes, ketones, pyridines, phenols, furans,
pyrazines, indenes, indoles and indanes. Because of its volatility, the
semi-volatile fraction includes a large proportion of components
which contribute to the flavor and aroma of smoke.
The chemical nature of mainstream smoke exiting the mouth end of
the cigarette changes as the smoke ages. This was demonstrated by
Vilcins and Lephardt (1975) for a number of gas phase components,
using Fourier transform infra red spectrometry to scan the undiluted
smoke immediately exiting the cigarette every 3.5 seconds (Fig.
12.11). In this example, the initial and rapid decrease in nitric oxide in
smoke is accompanied by an initial rapid build-up of nitrogen dioxide,
due to the atmospheric oxidation of nitric oxide. The nitrogen dioxide
concentration reaches a

 Page 413

Fig. 12.11
Aging of mainstream smoke,
concentration (c) vs time
(t, min) (Vilcins & Lephardt, 1975).
maximum just after a minute and then decreases. (These trends have
been confirmed by Cueto and Pryor (1994) using a similar technique).
The methanol concentration is constant for about 10 seconds and then
decreases slowly during the first minute, and its maximum rate of
decrease corresponds to the maximum level of nitrogen dioxide.
Similarly, no methyl nitrite is observed during the first 10 seconds, its
level in smoke is then observed to build up slowly before accelerating
after about one minute.
These observations suggest the following reaction sequence in smoke
as it ages during 2 to 3 minutes:
2NO + O2 ® 2NO2
CH3OH + 2NO2 ® CH3NO2 + HNO3
Thus, even in these conditions of undiluted smoke, no methyl nitrite
was observed in the smoke for about 10 seconds. Under human
smoking conditions, the cigarette smoke is diluted 50 to 100 fold by
the smoker during inhalation. For the trimolecular reactions involved,
an x-fold dilution of the smoke will result in an x3 fold reduction in
the rate of methyl nitrite formation. Thus, in the few seconds that the
diluted smoke is present inside the respiratory tract of the smoker, it is
highly unlikely that methyl nitrite will form.
The rate of oxidation of nitric oxide to nitrogen dioxide in mainstream
smoke has been determined in other studies (Sloan & Kiefer, 1969;
Williams, 1980; Borland, et al., 1985; Cueto & Pryor, 1994). The rate
constant measured in the gas phase of smoke is almost a magnitude
higher than for the pure gas phase conversion. The results of Borland,
et al. (1985) also show clearly that the oxidation of nitric oxide in the
smoke gas phase is faster than in whole smoke and have postulated
that free radicals present in the gas phase are catalyzing the oxidation.
They postulate that such free radicals must be quenched in whole
smoke.
As well as the changing chemical nature of smoke itself during
ageing, another problem that can occur is the formation of artifacts in
the collected smoke. Since there are many reactive species in smoke
there is potential for reaction in the smoke collection equipment to
produce chemicals that are not actually present in either fresh or aged
smoke. There are many examples of artifact changes occurring in
collected smoke (Dube & Green, 1982). For example, Schönherr, et
al. (1973) have shown that reactions, including polymerizations, occur
as cigarette smoke condensate and its solutions are stored over several
hours and days, even when stored at low temperatures which lead to
changes in composition. It is probable that such reactions would not
occur in actual smoke, casting doubt on the relevance of using old
smoke condensate in, for example, biological testing of tobacco
smoke. In a further example, cyanohydrins have been observed when
the mainstream smoke is collected in water but not when collected in
a cold trap at -79°C (Dube & Green, 1982). The cyanohydrins are
believed to be formed as artifacts by the reaction between hydrogen
cyanide and volatile carbonyl compounds:
RCHO + HCN ® RCH(OH)CN
where R = CH3·, C2H5·, n-C3H7· or i-C3H7·
As discussed in Section 10, Caldwell and Conner (1990) have
reported that artifactual formation of N-nitrosamines can occur on the
Cambridge pad part of the smoking trapping system of the analytical
method described by Hoffmann, et al. (1983) and Hecht, et al. (1983).
This is due to the nitrosation of amines trapped on the pad as tobacco
smoke is passed over them.
An effect of the analytical technique of a different nature is
encountered in an example of sidestream smoke analysis. Smoke
ammonia is found predominantly in the sidestream smoke. It is now
known that in the concentrated sidestream plume, ammonia and
formaldehyde react almost instantaneously to produce
hexamethylenetetramine (Borgerding, et al., 1984; Gonzá1ez &
Sarabia, 1992), which resides in the particulate phase of sidestream
smoke:

 Page 414

Because of the excess of ammonia, there is very little formaldehyde

actually present in sidestream smoke. However, during the collection
and work-up procedures for the analysis of the sidestream smoke, the
hexamethylenetetramine decomposes into ammonia and
formaldehyde, resulting in incorrectly high levels of formaldehyde
being quoted for sidestream. In contrast, in mainstream smoke
ammonia is present in low levels and so very little
hexamethylenetetramine is formed. In the presence of water vapor,
formaldehyde will exist as a hydrated form in mainstream smoke,
H2C(OH)2. In ETS, either the hexamethylenetetramine is never
formed or it decomposes to release formaldehyde and ammonia. In
ETS, formaldehyde is present in the free form or possibly the hydrated
form.
Thus, we see that there are some problems inherent in the chemical
analysis of smoke which in some circumstances can cause
uncertainties in their relevance to the smoker. Firstly, the chemical
nature of tobacco smoke changes as it ages so that the smoke that is
collected for analysis may no longer be representative of what the
smoker actually receives. Secondly, artifact formation can produce
substances during the analysis that are not present in fresh or aged
smoke or can produce artificially high levels of the analyte. These
effects should be borne in mind when considering data pertaining to
the chemical nature of smoke. However, it should also be borne in
mind that many, if not most, smoke components have been quantified
through tediously validated analytical procedures designed to
minimize artefact formation and ageing effects. Most of the
compounds reported in smoke are actually present and found in the
measured quantities.
Section 8
'Smoke pH'
Nicotine is dibasic and its pyridine ring is considerably less basic than
its pyrrolidine ring nitrogen:

Using acidbase equilibrium data in aqueous solution at 25°C, Morie

(1972) has calculated the fractions of free base or unprotonated (I),
monoprotonated (II) and diprotonated (III) nicotine at pH values from
one to 12. At pH 5.4, 100% of the nicotine is monoprotonated (II), as
the pH falls below 5.4 an increasing proportion of the diprotonated
(III) form is produced, and as the pH rises above 5.4 an increasing
proportion of the unprotonated form (I) is produced.
The monoprotonated (II) and diprotonated (III) forms of nicotine in
smoke will be bound ionically to the acids in smoke. The resulting
nicotine salts are involatile and will be contained exclusively within
the smoke aerosol particles. The free base or unprotonated form of
nicotine (I) has some volatility, although in mainstream smoke it also
resides substantially within the particulate phase, with only a trace in
the vapor phase.
The association with acids is a function of the alkalinity or acidity of
the smoke. Reference is often made in smoke chemistry papers to the
'pH of smoke' (e.g. Grob, 1961; Sensabaugh & Cundiff, 1967; Morie,
1972; Brunneman & Hoffmann, 1974) and this has sometimes been
interpreted in quantitative terms as to its effect on the form of nicotine
in smoke. Although there is an undoubted relationship between smoke
acidity and form of nicotine in smoke, this relationship is imprecise
and caution should be applied when attributing quantitative
significance to 'smoke pH'.
'Smoke pH' is measured in different ways in different studies, either as
the pH of an aqueous extract of the total particulate matter collected
from smoke or as the pH of an aqueous solution from whole smoke, or
when whole smoke is passed over an electrode covered with a thin
film of a buffer solution and connected to a pH meter. Thus quoted
ranges of 'smoke pH' depend on the exact measuring technique and
differ somewhat from study to study. However, typically quoted
ranges of 'smoke pH' together with the percentage of the unprotonated
(free base) form of nicotine in the smoke, calculated using acidbase
equilibrium data in aqueous solution (Morie, 1972), are:

Smoke 'Smoke Calculated %

pH' unprotonated
nicotine
Mainstream: flue- 5.06.0 01
cured cigarette
Mainstream: US- 5.56.5 0.33
blended cigarette
Mainstream: dark- 7.07.5 925
air cured cigarette
Mainstream: cigar 8.08.5 5080
Sidestream: 6.58.0 350
cigarette

 Page 415
However, in using the concept of pH, care has to be exercised in
interpreting its exact relevance to smoke at best 'smoke pH' has only
an indirect relationship, not quantifiable, to the form of nicotine in
smoke, pH refers to the hydrogen ion concentration in an aqueous
solution. Because nicotine is dibasic, it will be protonated by acids in
any medium and the extent of the protonation will depend on the
strength of the acids and their concentration. The pH of an aqueous
suspension of smoke is a measure of the acidity of the smoke in terms
of its ability to protonate a particular base (i.e. water) in a particular
medium (i.e. an aqueous solution). The acidity measured in this way
can only give an approximate indication of its ability to protonate a
different base (i.e. nicotine) in a different medium (i.e. smoke
particulate phase). The calculated '% unprotonated nicotine' figures
quoted above are, therefore, likely to be, at best, highly inaccurate.
The measured 'smoke pH' values are related to the sensory effects
experienced by the smoker, in particular the 'impact' of the smoke.
Impact is the localized sensation felt momentarily at the back of the
throat and is highly specific to nicotine. Tobacco blends high in sugar
content produce a more acidic smoke and generate smoke with
reduced impact, presumably due to the higher proportion of nicotine
in the salt form.
It has been speculated (International Agency for Research on Cancer
(IARC), 1985) that as the 'smoke pH' increases then the proportion of
nicotine in the vapor phase will increase because of the volatility of
the unprotonated nicotine. This is true in a general sense, but nicotine
in mainstream smoke is predominantly in the particulate phase. For
example, direct measurements (Bevan, 1995, unpublished results)
show that only 0.0007% of the mainstream nicotine from a fluecured
cigarette ('smoke pH' 6) is in the vapor phase while the equivalent
figure is 0.01% for mainstream nicotine from a fermented air-cured
cigarette ('smoke pH' 7.5). The equilibrium between vapor and
particulate phases will be established very rapidly. Therefore, when
smoke is in contact with a surface that has an affinity for nicotine, a
concentration gradient becomes established in which the nicotine is
transferred by isothermal distillation. The extent to which this occurs
depends on the equilibrium concentration in the vapor phase, hence on
the concentration of free base nicotine in the particulate phase, hence
on the acidity of the particulate phase and hence on the pH of an
aqueous suspension of the smoke ('smoke pH'). Thus nicotine in basic
solution (from cigar smoke, for example) is absorbed more rapidly
through the mucous membrane of the mouth than in more acidic
solution (from cigarette smoke) because of the higher concentrations
of unprotonated nicotine (Armitage & Turner, 1970; Gori, et al.,
1986). The question of whether sensory changes in smoke are
ultimately related to changes in the nicotine vapor-particulate
equilibrium or pH-driven changes in the kinetics of transport out of
deposited smoke particles is open to speculation.
Once smoke is dissolved in the aqueous fluids within the human body,
the form of nicotine (protonated versus unprotonated) will be
determined by the buffering capacity of the body fluid itself.
Section 9 Smoke Composition
The approximate chemical composition of the whole mainstream
smoke from an American-blended cigarette, smoked under the
standard smoking machine regime of one 35 cm3 puff of 2 seconds
duration, taken once per minute to a butt length of 23 mm, is depicted
in Table 12.4 (taken largely from Dube & Green, 1982, with similar
data also quoted in Keith & Tesh, 1965 & Norman, 1977). The
cigarette yielded 500 mg whole smoke, of which the particulate phase
weighed 22.5 mg, i.e. 4.5% by weight of the total smoke. For this
cigarette the nicotine yield is 1.5 mg (i.e. 0.3% of 500 mg) and the 'tar'
yield (weight of total particulate matter less the weight of nicotine and
particulate phase water) is 17.5 mg, i.e. it is a high 'tar' cigarette. Even
so, approximately 76% by weight of the mainstream smoke is ambient
air, almost 17% consists of the permanent gases CO2, CO, H2 and
CH4, and 2% is H2O. Many substances with molecular weights
between about 60 and 200, as well as water, are to some degree
partitioned between the vapor and particulate phases.
There are literally hundreds of papers on the composition of
mainstream smoke and the subject has been reviewed in detail (e.g.
Johnstone & Plimmer, 1959; Keith & Tesh, 1965; Wynder &
Hoffmann, 1967; Stedman, 1968; Green, 1977a; Green, 1977b; Hecht,
et al., 1977; Norman, 1977; IARC, 1985). Tobacco smoke consists of
many different types of chemicals, many of them present in extremely
low concentrations. Dube and Green (1982) estimated that there were
3875 organic chemical constituents in mainstream smoke, and the
numbers distributed among 15 classes of organic compounds are
listed in Table 12.5. This table lists a total of 4720 substances, which
is greater than the 3875 quoted because some molecules contain
multiple functional groups. Furthermore, the table does not include
the 100 or so metallic components, inorganic compounds and
inorganic ions which are also known to

 Page 416

Table 12.4
Approximate chemical composition of
whole mainstream smoke
(Dube & Green, 1982).
be present in smoke. Dube and Green (1982) estimated that there are
2550 substances in tobacco itself and 2470 substances that are unique
to smoke, i.e. formed by pyrolysis and pyrosynthesis reactions of
tobacco constituents in the burning cigarette. They estimated that
there are 1135 substances that are transferred unchanged from tobacco
to smoke and 1414 substances that are unique to tobacco, i.e. they are
not volatile enough to be transferred to smoke and/or decompose
when tobacco burns. About 500 of the smoke components are found
in the vapor phase and about 300 can be classed as semi-volatiles
(Norman, 1977).
It is sometimes implied from this very large number of compounds
present in mainstream smoke that tobacco smoke is a uniquely
complex substance. It is
Table 12.5 Approximate number of
compounds identified in some major
compound classes (Dube & Green,
1982).
Class Number
Amides, imides, lactams 237
Carboxylic acids 227
Lactones 150
Esters 474
Aldehydes 108
Ketones 521
Alcohols 379
Phenols 282
Amines 196
N-Heterocyclics 921
Hydrocarbons 755
Nitriles 106
Anhydrides 11
Carbohydrates 42
Ethers 311
Total 4720

complex. However, the large number of compounds reflect the intense

scrutiny of chemical investigators since the 1950s more than anything
else. Most of the thousands of substances found in smoke are present
in trace quantities. Other combustion mixtures and natural products
would be found to be equally complex if they were studied so
intensively.
In general, the same chemicals present in mainstream smoke are
present in sidestream smoke, though their relative yield per cigarette
is highly dependent on the compound considered. Some typical
mainstream yields and sidestream/mainstream ratios are shown in
Table 12.6, using data from a variety of sources (Neurath, et al., 1966;
Johnson, et al., 1973a; Browne, et al., 1980; Baker, 1981a; Kamstrup,
et al., 1981; Klus & Kuhn, 1982; Norman, et al., 1983; Sakuma, et al.,
1983; Sakuma, et al., 1984a; Sakuma, et al., 1984b; IARC, 1985;
NRC, 1986; Umemura, et al., 1986; Guerin, 1987; Eatough, et al.,
1990; Klus, 1990; Guerin, 1991; Guerin, et al., 1992). The data
selected for Table 12.6 refer to plain, unfiltered cigarettes of various
types, smoked under standard machine smoking conditions. Changing
the design parameters of the cigarettes, such as length, tobacco type
and paper characteristics, will alter the mainstream and sidestream
yields; hence the ranges quoted in Table 12.6. Cigarettes incorporating
filters and ventilated filters will generally have reduced mainstream
yields relative to their non filter cigarette counterparts, but similar

 Page 417
sidestream yields (e.g. Browne, et al., 1980; Chortyk & Schlotzhauer,
1989). Sidestream yields are primarily related to the weight of tobacco
burnt during smolder (Baker, 1984). Hence the sidestream/mainstream
ratios of cigarettes incorporating filters will be generally higher than
those quoted in Table 12.6.
When viewing the data in Table 12.6 it should be borne in mind that a
variety of different methods were used in the various studies to collect
the sidestream smoke for analysis (reviewed, for example, in Dube &
Green, 1982; McRae, 1990; Guerin, 1991). The different methods
involve using different shaped enclosures around the cigarette. These
various enclosures will affect the air flow conditions around the
cigarette to different extents, influencing burn rate during smolder and
yield of sidestream components. In addition, the different methods
have produced conflicting data on the distribution of sidestream
components between the two phases of smoke. Sidestream nicotine,
for example, has been reported to be almost entirely in the particulate
phase in some studies (Higgins, et al., 1984; Proctor, et al., 1988), and
50 to 80% in the vapor phase in other studies (Johnson, et al., 1973a;
Browne, et al., 1980; Sakuma, et al., 1984b). Similarly, sidestream
hydrogen cyanide has been reported to be exclusively in the
particulate phase in one study (Johnson, et al., 1973a) and 83 to 94%
in the vapor phase in another study (Norman, et al., 1983). Such
conflicting results are due to differences in the sidestream collection
procedures, the difficulty in unambiguously separating the phases in a
dense aerosol with a very high water content, and release of collected
substances from the Cambridge filter if it is heated by the sidestream
plume (resulting in a low collection efficiency and apparently low
particulate phase contribution). For these reasons, no attempt is made
in Table 12.6 to attribute the sidestream constituents to either phase.
In any case, unambiguously knowing the phase distribution of
constituents in fresh, undiluted sidestream smoke is not particularly
important because it changes as the smoke ages and dilutes to become
environmental tobacco smoke.
The substances included in Table 12.6 were selected from the
thousands of products in smoke as examples to illustrate the types of
product or product classes found and the range of
sidestream/mainstream ratios observed. The data in Table 12.6 do
show that there is a very wide range of sidestream/mainstream ratios,
varying from 0.03 to 0.1 for carbonyl sulfide to over 270 for
molecular nitrogen formed chemically from the tobacco. Of course,
one to four times more tobacco is burnt in smolder than during
puffing, depending on the cigarette parameters. Thus, some of the
ratios in Table 12.6 (e.g. phenol, n-butylamine and nicotine) simply
reflect the proportions of tobacco burnt to generate the two smoke
streams. However, there are many substances which distribute
themselves so predominantly to one stream or the other that the reason
cannot be due to differences in tobacco consumption. The reasons lie
in the different conditions of temperature and mass transfer rates
existing in the cigarette burning zone during smolder and puffing and
the exact mechanism by which different components are formed or
released from the tobacco. In general, alkaline and neutral constituents
are released to a greater degree in sidestream than mainstream. Acids,
phenols and phytosterols are released to a lesser or equal degree in
sidestream than mainstream.
Section 10 Generation of Specific Smoke Constituents
The following pages summarize the various routes that have been
postulated for formation of the smoke components listed in Table 12.6
in terms of the tobacco constituents from which they are generated.
Many of these suggested precursor-product routes have been obtained
from individual pyrolysis studies. As pointed out in Section 6, it
should be borne in mind that many of these pyrolysis studies have
used conditions of heating rate and residence time that bear little
resemblance to those inside the burning cigarette. The suggested
chemical routes below are the best indicators available at present, but
could be inaccurate. Furthermore, they are merely routes and, in
virtually all cases, the exact mechanisms occurring are obscure.
Carbon monoxide and dioxide are the major products in cigarette
smoke (Table 12.4) and are formed by both thermal decomposition
and combustion of many of the components of tobacco: starch,
celluloses, sugars, carboxylic acids, esters, amino acids etc. (Baker,
1981a). Just over half of each gas is formed by combustion (Baxter &
Hobbs, 1967; Johnson, et al., 1975; Baker, 1981a; Baker, 1984; Baker,
1987b). Within the burning cigarette about 30% of the carbon
monoxide is formed by thermal decomposition of tobacco
components, about 36% by combustion of tobacco and at least 23%
by carbonaceous reduction of carbon dioxide (Baker, 1987b):

Ventilation of the cigarette at the filter reduces the amount of air

drawn in through the burning zone,

 Page 418
Table 12.6 Some typical sidestream/mainstream (SS/MS)
yield ratios for plain cigarettes.
Substance MS yield SS/MS
Small molecules
Carbonyl sulphide 1842 µg 0.030.1
HCN 160500 µg 0.060.5
CO 1023 mg 2.54.7
Hydrazine 2040 µg 3
Methane 6001000 µg 3.14.8
Acetylene 2040 µg 0.82.5
Nitrogen oxides 100600 µg 413
CO2 2050 mg 811
H2O (gas phase) 314 mg 2430
NH3 50130 µg 40170
N2 (generated) <10 µg >270
Neutral heteroatom organics
Acetonitrile 160210 µg 35
Benzonitrile 56 µg 710
Acetamide 70100 µg 0.81.7
Methyl chloride 150600 µg 1.73.3
Aldehydes, ketones, alcohols
Acetaldehyde 0.51.2 mg 1.4
Propionaldehyde 175250 µg 2.42.8
Acetone 100250 µg 25
Acrolein 60100 µg 815
2-Butanone ~30 µg 2.94.3
2-Furaldehyde 1543 µg 4.97.4
Furfuryl alcohol 1865 µg 3.04.8
Cyclotene* 35 µg 610
Pyranone** 13150 µg 0.11.2
Phytosterols
b-Sitosterol 59 µg 0.5
Campesterol 43 µg 0.6
Cholesterol 22 µg 0.9
Phenols
Phenol 60140 µg 1.63.0
Cresols (o-, m-, p-) 1137 µg 1.01.4
Catecho l 100360 µg 0.60.9
Hydroquinone 110300 µg 0.71.0
Acids
Formic acid 210490 µg 1.41.6
Acetic acid 270810 µg 1.93.9
3-Methylvaleric acid 2060 µg 0.81.5
Lactic acid 60170 µg 0.50.7
Benzoic acid 1428 µg 0.71.0
Phenylacetic acid 1138 µg 0.60.8
Succinic acid 70140 µg 0.40.6
Glycolic acid 40130 µg 0.61.0

(table continued on next page)

 Page 419
Contd.
Table 12.6
Substance MS yield SS/MS
Amines, pyridines, alkaloids
Methylamine 1229 µg 4.26.4
n-Propylamine 1.63.4 µg 2.83.8
n-Butylamine 0.51.5 µg 2.24.0
Aniline 360 ng 30
Pyridine 1646 µg 6.520
3-Ethenylpyridine 1130 µg 2040
Methylpyrazine 25 µg 34
Pyrrole 1623 µg 914
Nicotine 0.82.3 mg 2.63.3
Myosmine 1333 µg 4.07.5
Nicotyrine 440 µg 514
Anatabine 220 µg 0.10.5
2,3'-Bipyridyl 1622 µg 23
Aza-arenes
Quinoline 0.52.0 µg 811
Isoquinoline 1.62.0 µg 2.55
Benzo[h]quinoline 10 ng 10
Indole 1638 µg 2.13.4
Hydrocarbons
Isoprene 3301100 µg 1319
Benzene 3668 µg 510
Toluene 100200 µg 68
Limonene 1550 µg 412
Neophytadiene 66230 µg 12
Polynuclear aromatic hydrocarbons
Naphthalene 2.6 µg 17
Pyrene 45140 ng 211
Benzo[a]pyrene 940 ng 220
Anthracene 24 ng 30
Phenanthrene 77 ng 230
Fluoranthene 60150 ng 11
Nitrosamines***
N-nitrosodimethylamine 1040 ng 1050
N-nitrosodiethylamine nd25 ng 335
N-nitrosopyrrolidine 630 ng 630
N-nitrosodiethanolamine 070 ng 1.2
N'-nitrosonornicotine 0.23 µg 0.53
NNK**** 0.11 µg 14
N'-nitrosoanatabine 0.35 µg 0.31
Inorganic constituents
Cadmium 100 ng 47
Nickel 2080 ng 0.230
Zinc 60 ng 0.27
* 2-Hydroxy-3-methyl-2-cyclopentanone.
** 5,6-Dihydro-3,5-dihydroxy-2-methyl-4H-pryan-4-one.
*** Much of the data in the literature on the smoke levels of
volatile and tobacco specific nitrosamines may be in error,
due to artifact formation on the Cambridge pad part of the
smoke collection procedure (Caldwell & Conner, 1990).
**** 'Nitrosonornicotine ketone' or 4-(methylnitrosamino)-l-
(3-pyridyl)-1-butanone.
nd = not detected.

 Page 420
reduces the temperature in a puff and reduces the amount of carbon
dioxide converted to carbon monoxide (Baker, 1981a). Consequently,
the mainstream CO2/CO ratio increases as the ventilation of the
cigarette filter is increased (Browne, et al., 1980). However, the
sidestream CO2/CO ratio is independent of filter ventilation, since
ventilation has only a minor effect on burning zone temperature
during the smolder period when most of the carbon oxides are
generated. There is relatively more carbon dioxide than carbon
monoxide in sidestream smoke (CO2/CO is 8 to 13 in sidestream and
2 to 6 in mainstream smoke). This is due predominantly to the lesser
occurrence of the carbonaceous reduction of carbon dioxide since the
burning zone surface temperatures of less than 600°C during smolder
are too low for the reduction to occur.
Gas-phase water is a major combustion product that rivals carbon
monoxide and carbon dioxide on a mass basis. However, unlike the
carbon oxides, gas-phase water is delivered predominantly to the
sidestream, and the incorporation of atmospheric oxygen in
sidestream water is much higher than in mainstream water (Johnson,
et al., 1975; Kornfeld, et al., 1979). This suggests that gas-phase
water is largely a secondary product formed via pyrolytically
generated hydrogen which diffuses into the sidestream plume where it
is oxidized to water by oxygen near the surface of the burning zone at
temperatures of 300500°C (Baker, 1982).
The gas-phase hydrocarbons such as methane, ethane, propane,
ethylene, propylene and acetylene are formed from the thermal
decomposition of a whole range of leaf constituents, paraffins,
triglycerides, aliphatic acids, amino acids, aldehydes and esters
(Chortyk & Schlotzhauer, 1973; Schlotzhauer & Chortyk, 1987)
generally at temperatures between about 300 and 550°C (Burton &
Childs, 1975; Baker, 1976a; Burton & Childs, 1977).
A portion of the hydrazine (NH2.NH2) present in smoke is transferred
from tobacco during smoking where it may have been formed by
reduction of nitrate or as an impurity from the plant sucker growth
inhibitor maleic hydrazide (Liu, et al., 1974). The results from
pyrolysis experiments have also suggested that hydrazine in smoke
can be formed from the thermal decomposition of amino acids,
protein and maleic hydrazide (Liu, et al., 1974).
Pyrolysis studies by Patterson, et al. (1976) show that carbonyl
sulphide (COS) is formed from the thermal decomposition of sulfur
containing amino acids such as cysteine, cystine, homocystine,
methionine, methionine sulfone and methionine sulfoxide at 850°C.
Other pyrolysis studies of the amino acids at 150 to 400°C (Fujimaki,
et al., 1969; Kato, et al., 1973) did not report carbonyl sulfide among
the reaction products. Thus, since temperatures in the order of 850°C
are apparently required to generate the carbonyl sulfide from the
amino acids, and since these temperatures are only reached during
puffing and not the interpuff smolder period, carbonyl sulfide is
delivered almost exclusively to the mainstream smoke (Kamstrup, et
al., 1981).
Fresh mainstream smoke exiting the butt end of the cigarette contains
100 to 600 µg nitric oxide (NO), and, as discussed in Section 7 above,
this is oxidized to nitrogen dioxide (NO2) if the smoke is allowed to
age for a few seconds or more. A principal source of nitric oxide is
thermal decomposition of nitrates in the tobacco (Johnson, et al.,
1973b; Hardy & Hobbs, 1977; Hecht, et al., 1977; Williams, 1980;
Brunnemann & Hoffmann, 1982; Norman, et al., 1983; Umemura, et
al., 1986). A linear relationship exists between mainstream nitric
oxide yield and tobacco nitrate level, with an extrapolated residual
level of 3 to 10 µg mainstream nitric oxide per puff at zero nitrate
(Williams, 1980; Norman, et al., 1983; Umemura, et al., 1986).
Similarly, there is a distinct relationship between sidestream nitric
oxide yield and tobacco nitrate, with an extrapolated residual level of
60 to 100 µg sidestream nitric oxide per minute at zero nitrate
(Norman, et al., 1983; Umemura, et al., 1986). Clearly, there must be
other sources of nitric oxide. By measuring nitric oxide yields from
nitrogen-free cellulose cigarettes, two studies have shown
independently that the residual mainstream nitric oxide level must be
due to formation by oxidation of atmospheric nitrogen (Norman, et
al., 1983; Umemura, et al., 1986). However, both studies have also
shown that nitrogen oxidation can only account for about 15% of the
residual sidestream nitric oxide yield. Umemura, et al. (1986)
concluded that a considerable amount of sidestream nitric oxide is
formed by the oxidation of organic nitrogenous compounds such as
amino acids and protein.
Ammonia is delivered largely to sidestream smoke and is formed from
the reduction of nitrates and pyrolysis of glycine (johnson, et al.,
1973b). Hydrogen cyanide is also formed from nitrates, glycine,
proline and aminodicarboxylic acids (Johnson & Kang, 1971;
Johnson, et al., 1973b; Norman, et al., 1983), but unlike ammonia, it
is delivered predominantly to mainstream smoke. The reactions that
form hydrogen cyanide must only occur at the relatively high
temperatures generated during the puff, whereas the reactions of the
nitrates and glycine that form ammonia must occur at

 Page 421
the lower temperatures that are experienced in the interpuff smolder
period. Johnson, et al. (1973b) have shown that the conversion of
nitrate nitrogen to ammonia during smoking is highly efficient. For
example, 25% of the nitrogen added as calcium nitrate was converted
to sidestream ammonia. They postulated that much of the ammonia
initially formed from the nitrate would be converted to other smoke
components.
In another paper, Johnson, et al. (1973c) have shown that molecular
nitrogen generated in the burning cigarette is delivered almost
exclusively to the sidestream. Their results using 15N-labeled nitrate
salts and glycine added to cigarettes indicate that molecular nitrogen
is formed from both nitrates and glycine, and they suggest it is formed
via ammonia. This conversion could occur as the ammonia diffuses
out of the oxygen deficient burning zone to the sidestream, where it
reacts with oxygen:

In contrast, Hardy and Hobbs (1977) have also added 15N-labeled

nitrate to tobacco and found that the generated nitrogen was not
derived from the nitrate nitrogen. This is another illustration of the
complexities of a burning cigarette and how unresolved questions still
remain on the chemistry involved.
By analyzing the mainstream products of cigarettes with added 15N-
labeled nitrates (potassium, sodium or calcium) and glycine, Johnson,
et al. (1973b) have shown that 15N was recovered in formamide,
acetamide and propionamide in all cases. They postulated that the
amides were formed via the reaction of intermediary ammonia with
other smoke components, the ammonia being generated from the
reduction of nitrates and pyrolysis of glycine.
In the same study (Johnson, et al., 1973b), 15N was also recovered in
the acetonitrile from cigarettes with added nitrate, but not with added
glycine. The isolation of acetonitrile-14C in the gas phase of
mainstream smoke of cigarettes treated with D-glucose-14C or D-
sucrose-14C indicates a reaction during smoking between sugars or
their degradation products and N-containing components of tobacco or
smoke (Gager, et al., 1971). The results from direct pyrolysis studies
at temperatures above 700°C suggest that acetonitrile is also formed
from pyrosynthetic reactions involving amino acids, peptides and
alkaloids (Johnson & Kang, 1971; Johnson, et al., 1973d; Schmeltz &
Hoffmann, 1977).
Several studies have shown that benzonitriles and naphthonitriles are
produced during the pyrolysis of lysine, leucine, tryptophan,
phenylalanine, maleic hydrazide and N,N-dimethyldodecylamine
(Brunnemann & Hoffmann, 1982).
A proportion of the aldehydes and ketones in smoke is transferred
from tobacco by distillation (Stedman, 1968) where they are formed
by nonenzymatic browning reactions (Leffingwell, 1976). However,
the largest proportion is formed by the pyrolysis of a number of
tobacco constituents, in particular celluloses, sugars, pectins, tobacco
lipids and waxes, protein, amino acids and triglycerides (Brunnemann
& Hoffmann, 1982; Chortyk & Schlotzhauer, 1973; Miyake &
Shibamoto, 1995; Schlotzhauer & Chortyk, 1987).
Tobacco smoke contains a variety of alcohols. In addition to allyl
alcohol, benzyl alcohol, and 2phenylethyl alcohol (IARC, 1985),
tobacco smoke and tobacco contain the straight-chained primary
alcohols ranging from methanol to 1-tetracosanol [CH3.
(CH2)22.CH2OH] plus 1-octacosanol [CH3.(CH2)26.CH2OH].
Primary alcohols ranging from 1-hexadecanol to 1-dotriacontanol
have been identified as esters with long-chained aliphatic acids in
tobacco and tobacco smoke (Arrendale, et al., 1988). The primary
alcohols arise in smoke by direct transfer of the unbound alcohol from
tobacco or by hydrolysis of long-chained esters to the alcohol and
acid. Other alcohols in tobacco and smoke include various terpenoid
alcohols such as phytol (C20) and solanesol (C45), the latter now
considered a good marker for ETS (Ogden & Maiolo, 1989).
Like all higher plants, tobacco contains several phytosterols,
particularly representative examples being b-sitosterol, campesterol
and cholesterol (IARC, 1985). Total phytosterols constitute about 10%
of the weight of the 'terpenoid fraction' of tobacco and about 0.2% of
the tobacco itself (Schmeltz, et al., 1975). They are present in tobacco
in the unbound form and as glycosides and fatty acid esters. About 20
to 25% of the phytosterols in tobacco are transferred intact to smoke
(Schmeltz, et al., 1975) with sidestream/mainstream ratios varying
between 0.5 and 0.9 (Table 12.6). The transfer is either by distillation
of the free sterol or by distillation of sterols freed by hydrolytic
degradation of the esters and glycosides. The remainder of the tobacco
phytosterols, both free and bound, undergo extensive degradation to
yield a wide variety of products, including gaseous hydrocarbons and
polynuclear aromatic hydrocarbons (e.g. Schmeltz, et al., 1975;
Schlotzhauer & Chortyk, 1987).
A total of over 300 phenols has been reported in smoke ranging from
simple volatile monophenols to

 Page 422
polyphenols. The polyphenols transfer directly from tobacco by
distillation. Leaf biopolymers such as lignin and the celluloses are the
principal precursors of monophenols in tobacco smoke, although
pyrolysis of other polysaccharides, sugars, protein, amino acids as
well as leaf polyphenols such as chlorogenic acid generate a wide
variety of phenols (Stedman, 1968; Wakeham, 1972; Chortyk &
Schlotzhauer, 1973; Schlotzhauer & Chortyk, 1987). The volatile
phenols are selectively removed from mainstream smoke by passage
through the plasticized cellulose acetate filter. This was the first
reported example of selective filtration by charcoal-free filters
(Williamson, et al., 1965). A similar phenomenon was subsequently
observed with the volatile N-nitrosamines (Brunnemann, et al., 1977;
Hoffmann, et al., 1980).
Tobacco smoke contains a variety of volatile carboxylic acids (C1 to
C5), longchain fatty acids (C6 to C22), hydroxycarboxylic acids,
dicarboxylic acids and benzoic acids (IARC, 1985). The volatile acids
are transferred to smoke from tobacco (Stedman, 1968) as well as
being formed by pyrolysis of substances in tobacco, including esters,
triglycerides, lactic acid salts and the biopolymers starch, pectins and
celluloses (Chortyk & Schlotzhauer, 1973). The characteristic flavor
of the smoke of Oriental tobacco is associated with concentrations of
3-methylvaleric acid (Stedman, 1968; Elmenhorst, 1972; Roberts,
1988). The longchain saturated and unsaturated fatty acids originate in
tobacco and transfer to smoke (Hoffmann & Woziwodzki, 1968;
Severson, et al., 1978).
Nearly 200 aliphatic and aromatic amines have been detected in both
processed tobacco and smoke, indicating that a proportion of those
present in smoke is due to transfer during smoking (Schmeltz &
Hoffmann, 1977; Heckman & Best, 1931). They are formed by
thermal degradation of alkaloids, protein and amino acids in tobacco
(Chortyk & Schlotzhauer, 1973).
Some 50 pyrazines, 55 pyrroles and over 350 pyridines have been
identified in tobacco smoke (Heckman & Best, 1981; Heckman, et al.,
1981). There are, in fact, more pyridines in smoke than any other class
of heterocyclic compound. These semi-volatile five and six membered
ring N-heterocyclic compounds contribute significantly to the flavor
of tobacco smoke and especially to that of cigars and pipes
(Leffingwell, 1976; Heckman, et al., 1981; Roberts, 1988). A
proportion of these compounds transfer to smoke by direct transfer
from tobacco, where they were formed by nonenzymatic browning
reactions involving amino acids and saccharides (Leffingwell, 1976).
The N-heterocyclic compounds are also formed by pyrolytic reactions
involving nicotine and other alkaloids, amino acids, protein, nitrates,
saccharides and Amadori compounds in tobacco which are formed by
condensation reactions of sugars and amino acids in the tobacco
(Chortyk & Schlotzhauer, 1973; Leffingwell, 1976; Hecht, et al.,
1977; Schmeltz & Hoffmann, 1977; Schmeltz, et al., 1979). Osdene
(1976) has suggested that in the cigarette burning zone, pyrosynthetic
reactions of ammonia with hydroxymethyl-tetrahydrofuran and
acrolein, for example, form N-heterocyclic compounds:

The structures of some of the common tobacco alkaloids found in

smoke are given in Fig. 12.12 together with typical mainstream yields
for a plain, nonfilter cigarette (IARC, 1985; Guerin, 1991). Nicotine
accounts for 85 to 90% of the total alkaloids in smoke. The fate of
nicotine when the cigarette is smoked has been discussed previously
in Section 6. Most of it transfers intact to smoke with a relatively
small amount of decomposition. Much less information is available
for the behavior of the other tobacco alkaloids in a burning cigarette.
It is known that nornicotine transfers less readily to mainstream
smoke than nicotine (Hecht, et al., 1977) and nicotine and nornicotine
produce broadly similar substances when pyrolyzed (Kuhn, 1965).
Aza-arenes are fused ring N-heterocyclic compounds. More than 230
have been reported in tobacco and smoke and contain two, three, four
and five fused rings (Klimisch & Beiss, 1976; Schmeltz & Hoffmann,
1977; Snook, 1978; Heckman & Best, 1981; Snook, et al., 1981;
Grimmer, et al., 1987; Kamata, et al., 1992; Motohashi, et al.,
1993) a few are shown in Figure 12.12 with typical mainstream
yields from a plain cigarette. Most of these compounds are found in
smoke with, at most, only very small levels present in tobacco,
indicating that they are formed predominantly by pyrolysis and
pyrosynthetic reactions. They are formed from amino acids, protein,
possibly nicotine and pyr-

 Page 423

Fig. 12.12
Structures of some smoke
 components (and typical
mainstream yields from plain cigarettes).
role, and reactions of amino acids (especially tryptophan) with
aldehydes (Chortyk & Schlotzhauer, 1973; Schmeltz & Hoffmann,
1977).
The analysis of the small quantities of azaarenes in smoke is complex,
and there are contradictory findings in the literature on the presence of
certain azaarenes in tobacco smoke condensate and the condensate
from tobacco pyrolysis experiments. Starting with 250 g of cigarette
smoke condensate, Van Durren, et al. (1960) claimed to have
identified the three pentacyclic azaarenes depicted in Table 12.7 and
quoted mainstream deliveries of 0.1 to 3 ng per cigarette. One of the
azaarenes was also reported in unpublished data (Candeli, et al., 1963)
cited in Wynder and Hoffmann (1967). However, these identifications
have not been confirmed in seven other studies conducted in Japan,
Germany and the USA; all used analytical techniques that would have
been reasonably expected to detect the substances had they been
present (Table 12.7, Rodgman, 1998). In spite of this lack of
confirmation of the presence of these three azaarenes in tobacco
smoke, they were still cited as ''significant tobacco smoke tumorigens'
by Hoffmann and Hecht (1990).
Tobacco smoke contains a large variety of hydrocarbon compounds.
More than 100 alkanes, 150 alkenes and 55 alicyclic hydrocarbons
have been identified in tobacco smoke (Newell, et al., 1978; Heckman
& Best, 1981). The yields decrease drastically as the number of
carbon atoms in the molecule increases, and this is generally true for
homologous series of many tobacco smoke components unless there is
a unique precursor for a specific member of the series (Norman,
1977). The average transfer rate of C25 to C33 hydrocarbons from
tobacco to mainstream smoke is 25%, for a plain, nonfilter cigarette
under standard smoking conditions (Severson, et al., 1978). They are
transferred from the tobacco waxes which contain alkanes, alkenes,
alcohols, carboxylic acids, esters, aldehydes, ketones and alkaloids.
The hydrocarbons transfer either structurally intact or fragmented into
alkanes and alkenes of chain lengths shorter than C25. Hundreds of
isoprenoids, including tobacco-specific compounds, have been
identified in tobacco and smoke (e.g. Newell, et al., 1978; Enzell &
Wahlberg, 1980; Wahlberg & Enzell, 1984). The typical aroma of
tobacco leaves generated during post-harvest is due chiefly to
isoprenoids. The most prevalent acyclic isoprenoids include solanesol,
phytone and neophytadiene, and the predominant cyclic isoprenoids
include cembranoids and carotenes. Solanesol, phytol and
neophytadiene are transferred unchanged from tobacco into
mainstream smoke, as are several cembranoids and carotenes. One of
the pyrolytic decomposition products of most isoprenoids is isoprene
(2-methyl-l,3-butadiene) formed between about 300 and 520°C in the
burning zone (Burton & Childs, 1975; Burton & Childs, 1977), and its
level in mainstream smoke is relatively high, typically 330 to 1100 µg
for a plain, nonfilter cigarette (Brunnemann, et al., 1990). Its pyrolytic
formation temperature is such that it will be generated during smolder,
hence its relatively high SS/MS ratio (Table 12.6).
More than 75 monocyclic aromatic hydrocarbons such as benzene and
toluene are formed from the pyrolysis of amino acids, fatty acids,
cinnamic acid,

 Page 424
Table 12.7 Identification of three pentacyclic aza-arenes in cigarette
smoke condensate (CSC) or tobacco pyrolysate (Pyr).
Aza-arene A Aza-arene B Aza-arene C
Investigators Pyr CSC Pyr CSC Pry CSC
Van Duuren, et al. (1960) Yes Yes Yes Yes No Yes
Candeli, et al. (1963) NE No NE Yes NE NE
Kaburaki, et al. (1970) No NE No NE NE NE
Schmeltz, et al. (1972) No NE No NE No NE
Schmeltz, et al. (1979) No No No No No No
Snook (1978) NE No NE No NE No
Snook, et al. (1981) NE No NE No NE No
Grimmer, et al. (1987) NE No NE No NE No
Kamata, et al. (1992) NE No NE No NE NE
Yes = compound identified.
No = compound not found.
NE = substrate not studied or not examined for compound.

Aza-arene Structure Code

Dibenz[a,h]acridine A
Dibenz[a,j]acridine B
7H-Dibenzo[c,g]carbazole C

sugars and paraffins (Chortyk & Schlotzhauer, 1973; Schlotzhauer &

Chortyk, 1987), precursors with an aromatic or cyclohexane ring and
pyrosynthesis from primary hydrocarbon radicals (Badger, 1962).
Polynuclear aromatic hydrocarbons have been determined in tobacco
smoke in many studies. In one large series of studies, Snook and co-
workers (Severson, et al., 1976; Snook, et al., 1976; Snook, et al.,
1977; Arrendale, et al., 1980) smoked approximately 40 000
cigarettes to obtain 1 kg smoke condensate. Various extraction and
separation techniques were then employed, and over 300 individual
polynuclear aromatic hydrocarbons were identified. The structures of
some of the parent polynuclear aromatic hydrocarbons found in the
smoke condensate are shown in Fig. 12.12 together with typical
mainstream yields from a plain cigarette (Norman, 1977; Arrendale, et
al., 1980; Guerin, 1991). In terms of numbers of compounds, the

 Page 425
list of polynuclear aromatic hydrocarbons is dominated by compounds
with methyl side chains since there are many permutations where the
side chains can be attached. Thus, for pyrene alone there are three
possible monomethylpyrenes, 15 dimethylpyrenes, 32
trimethylpyrenes, and so on, for a total of 287 possible methylpyrenes,
although not all of them have so far been detected in smoke (Norman,
1977). There are at least 80 naphthalenes present in smoke (Schmehz,
et al., 1976; Arrendale, et al., 1980). The polynuclear aromatic
hydrocarbons are formed by pyrolysis and pyrosynthetic reactions of
terpenes, phytosterols such as stigmasterol, paraffins, sugars, amino
acids, celluloses and many other tobacco components (Chortyk &
Schlotzhauer, 1973; Schmeltz, et al., 1976; Scholtzhauer & Chortyk,
1987), and reactions involving primary hydrocarbon radicals (Badger,
1962).
Several independent studies have determined the formation of
hydrocarbons as a function of temperature during the pyrolysis of
tobacco (Burton & Childs, 1975; Baker, 1976a; Burton & Childs,
1977) and substances found in tobacco leaf such as n-C25H52, phytol,
neophytadiene, n-dotriacontane and stigmasterol (Badger, et al., 1965;
Scholtzhauer, et al., 1970; Schmeltz, et al., 1976; Lam, et al., 1985).
The results all generally show that at temperatures of about 400° to
600°C series of normal alkanes and alkenes are generated. At
temperatures above 500°C, benzene and alkylbenzenes are formed. At
temperatures of about 700°C, naphthalenes are formed, and above
800°C, polynuclear aromatic hydrocarbons are generated. Although
the relevance of pyrolysis experiments to smoke generation needs to
be treated with caution, these independent results are generally
consistent and indicative of the temperature zones involved in
hydrocarbon formation.
Tobacco smoke contains two general types of nitrosamines:
(1) tobacco-specific nitrosamines (e.g. N´-nitrosonornicotine (NNN))
that are derived from tobacco alkaloids and are found only in tobacco
and the particulate phase of tobacco smoke; they are non-volatile; and
(2) nontobacco-specific nitrosamines that are also found in other
systems (e.g. N-nitrosomethylamine and N-nitrosopyrrolidine); they
are often called 'volatile nitrosamines' and are present in the vapor or
semi-volatile phase of mainstream smoke.
It is generally accepted that tobacco-specific nitrosamines are not
present in freshly harvested green tobacco (Brunnemann & Hoffmann,
1991; Wiernik, et al., 1995). They are formed during tobacco curing
by nitrosation of tobacco alkaloids (Fig. 12.13). NNN, N-
nitrosoanabasine (NAB) and N´-nitrosoanatabine (NAT) are formed
predominantly by N-nitrosation of the corresponding secondary amine
(minor alkaloid). Some NNN can also be formed by nitrosation of the
tertiary amine nicotine, following loss of the methyl group (IARC,
1985; Brunnemann & Hoffmann, 1991; Wiernik, et al., 1995). 4-
(Methylnitrosemine)-l-(3-pyridyl)-l-butanone (NNK) can only be
formed from nicotine by oxidative N-nitrosation following ring
opening of the pyrrolidine ring. The nitrosating agent is nitrite,
derived from tobacco nitrate by the action of bacteria and tobacco
enzymes during curing (Parsons, et al., 1986; Wiernik, et al., 1995).
Thus, the amount of nitrates in tobacco has an important effect on the
level of tobacco-specific nitrosamines. The final level of tobacco-
specific nitrosamines in tobacco is dependent on these factors:
tobacco variety and agronomic conditions which affect the initial
alkaloid and nitrate concentrations; curing conditions during which
nitrosation of the alkaloids occurs; and storing and processing
conditions where further nitrosation can occur.
 Fig. 12.13
Formation of tobacco-specific nitrosamines.
When tobacco burns during cigarette smoking, the tobacco-specific
nitrosamines can transfer to smoke as well as decompose thermally,
and additional amounts of the nitrosamines can be formed
pyrosynthetically. Based on the use of radiotracer techniques,
Hoffmann and co-workers (Hoffmann, et al., 1977; Adams, et al.,
1983) concluded that 40 to 46% of the N´-nitrosonornicotine in
mainstream smoke is transferred directly from the tobacco, with the
remainder being formed by pyrosynthesis during smoking. With
NNK, they estimated that only 26 to 37% is transferred from tobacco
while the rest is formed by pyrosynthesis. On the other hand, Fischer
and co-workers (1990) have concluded from studies of cigarettes
spiked with nitrosamine precursors (nitrate and nicotine) that the

 Page 426
pyrosynthesis of N´-nitrosonornicotine does not occur during
smoking. They concluded that the pyrosynthesis of NNK during
smoking is very unlikely, especially if the tobacco nitrate levels are
low. These conflicting conclusions illustrate the general complexity of
the chemical processes involved and the difficulty in designing
experiments that give definitive results without artifact effects.
The only nonvolatile nitrosamine identified in tobacco and smoke that
is not tobacco specific is N-nitrosodiethanolamine. Its presence has
been related to treating the growing tobacco with the sucker growth
inhibitor maleic hydrazide used as the diethanolamine salt. Tobacco
grown without the use of this agrochemical and smoke generated from
it do not contain N-nitrosodiethanolamine (IARC, 1985; Brunnemann
& Hoffmann, 1991). Maleic hydrazide used for agrochemical
application is now formulated as the potassium salt, so this particular
nitrosamine is no longer found in tobacco or smoke.
The levels of the nontobacco-specific, or volatile, nitrosamines in
mainstream smoke are much lower than those of the tobacco-specific
nitrosamines (Table 12.6), and less work has been done on
determining routes that lead to their formation. Although minute
amounts are found in processed tobacco and Could transfer to smoke,
it is generally believed that their main source in smoke is
pyrosynthetic formation (IARC, 1985; Brunnemann & Hoffmann,
1991). By spiking a cigarette in successive model experiments with
nitrate, secondary amines, nitrosamines, nonvolatile N-nitrosoamino
acids and amino acids and analyzing the resultant mainstream smoke,
Tricker and Preussmann (1992) have calculated the relative
importance of the following pathways for the formation of N-
nitrosopyrrolidine:
(a) 1 1/2% produced by direct transfer from tobacco
(b) 37% produced by pyrosynthetic nitrosation of the analogous
secondary amine (pyrrolidine):

(c) 10% produced by thermal decarboxylation of the analogous

nonvolatile N-nitrosoamino acid (N-nitrosoproline) which is present in
tobacco as a result of nitrosation of the corresponding amino acid:

(d) 52% produced by nitrosation and decarboxylaion of the

corresponding amino acid (proline) and/or tobacco protein:

Steps (b), (c) and (d) are all pyrosynthetic routes. The quoted
proportions of these routes probably do not have general applicability
to other volatile nitrosamines because of the different proportions of
the specific precursors. The nitrosating agent in (b) is believed to be
N2O3, formed by reaction of NO with NO2 (Neurath, et al., 1976;
Brunnemann & Hoffmann, 1991). Thus amino acids and protein in
tobacco which can generate amines on burning and nitrate which can
generate nitrogen oxides are important precursors of the volatile
nitrosamines (Hecht, et al., 1977; Brunnemann & Hoffmann, 1982;
Ishiguro, et al., 1984; IARC, 1985; Brunnemann & Hoffmann, 1991).
The sidestream/mainstream ratios of volatile nitrosamines are
relatively high (Table 12.6), presumably due to their pyrosynthetic
formation occurring at the relatively low temperatures encountered
during the interpuff smolder periods arid the relatively high levels of
NO and NO2 in the sidestream. The volatile nitrosamines are also
selectively removed (i.e. removed relatively more than total
particulate matter) from mainstream smoke by passage through a
plasticized cellulose acetate filter tip (Brunnemann, et al., 1977;
Hoffmann, et al., 1980). Since the sidestream yields of cigarettes will
be relatively unaffected by the presence of a filter, some filtered
cigarettes have sidestream/mainstream ratios for N-
nitrosodimethylamine, for example, approaching 100 (IARC, 1985)
largely because of the very low levels in mainstream smoke.
For many of the secondary amines present in tobacco and smoke there
are corresponding N-nitrosamines. However, in some instances N-
nitrosamines have been identified but not their corresponding amines
which, by necessity, must be present. Examples are
Nnitrosoethylisopropylamine and N-nitrosoethylisobutylamine
(Rodgman, 1995, personal communication). Also, there are at least 30
secondary amines in tobacco and smoke for which no corresponding
N-nitrosamines have been identified. For example, various
alkylpiperidines have been identified in tobacco smoke but not their
corresponding N-nitrosamines. However, they have been prepared
synthetically in nontobacco-related studies (Rodgman, 1995, personal
communication).
It should be noted that Caldwell and Conner (1990) published a paper
which indicated that artifactual

 Page 427
formation of both volatile and tobacco-specific N-nitrosamines could
occur on the Cambridge pad part of the smoke trapping system of the
analytical system described by Hoffmann and co-workers (Hecht, et
al., 1983; Hoffmann, et al., 1983) with both mainstream and
sidestream smoke collection. This was due to nitrosation of amines
trapped on the pad as tobacco smoke is passed over them. Caldwell
and Conner (1990) reported artifact formation producing
overestimates of mainstream tobacco specific nitrosamines of between
19% (for NNN), 83% (for NAB) and 380% for N-nitrosopyrrolidine.
For sidestream smoke they reported overestimates of between 56 and
94%. They found that when the Cambridge filter was treated with
ascorbic acid before smoke collection artifact formation was inhibited,
as has been observed by other workers (Brunnemann & Hoffmann,
1991). Thus, the quoted mainstream yields in Table 12.6 may be high
and the sidestream/mainstream ratios may be in error.
Like all plant materials, tobacco contains minerals and other inorganic
constituents. Upon combustion, most metals remain in the ash,
although a small proportion may form volatile compounds or be
carried in microfragments of ash and thus appear in smoke. Some 30
metals and other elements have been identified in tobacco smoke and
quantitative data are available for some of them (Allen & Vickroy,
1976; Jenkins, et al., 1985;Jenkins, 1990). The sidestream/mainstream
ratios quoted in Table 12.6 for cadmium, nickel and zinc are very
variable. However, Klus (1990) has pointed out that nearly all the
sidestream data on cadmium and other metals in smoke have been
calculated from the amount of the elements in the tobacco burnt, in
the mainstream smoke and in the remaining butt. No direct sidestream
determinations have been undertaken.
In addition to the relatively stable molecular products in tobacco
smoke discussed above, cigarette smoke also contains free radicals.
Pryor and co-workers have reported that cigarette smoke radicals are
of two distinct types: long-lived radicals present in the particulate
phase and short-lived radicals present in the vapor phase (Pryor, et al.,
1983a; Church & Pryor, 1985). The long-lived radicals are observed
directly in both the mainstream and sidestream smoke condensate
deposited on a Cambridge filter by means of electron spin resonance
(ESR) spectroscopy. There were approximately 1 × 1017 radicals per
gram of tar or 4 × 1014 radicals per puff. The ESR signals from
condensate from both smoke streams remained unchanged over
several days. Pryor and co-workers postulated that the principal long-
lived free radicals in smoke condensate are polymers that contain a
semi-quinone radical associated with quinone and hydroquinone
groups (Church & Pryor, 1985; Pryor, et al., 1983a; Pryor, et al.,
1983b).

The quinone and hydroquinone species can readily interconvert by

hydrogen atom exchange to produce the charge-transfer complexes.
The short-lived radicals in the vapor phase of smoke cannot be
observed by direct ESR spectroscopy and can be studied only by an
ESR spin trapping technique. This involves trapping the short-lived
radical with a compound that is more stable than the radical that was
trapped and can consequently be detected by ESR. Using this
technique, Pryor and co-workers (Pryor, et al., 1983a; Church &
Pryor, 1985) have shown that alkyl and alkoxyl radicals are present in
the smoke vapor phase (i.e. smoke passed through a Cambridge filter
and subsequently bubbled through the spin trapping reagent).
Approximately 1 × 1016 radicals per cigarette are present in both the
mainstream and sidestream smoke vapor phases, or 5 × 1014 per puff.
These types of radicals are known to have lifetimes of fractions of a
second. However, Pryor and co-workers (Pryor, et al., 1983a; Church
& Pryor, 1985) observed that in cigarette smoke the radicals have
apparent lifetimes in excess of 5 minutes, and their levels build up in
smoke during the first minute of ageing (Fig. 12.14). They have
postulated a steady-state mechanism for the continuous formation and
destruction of the radicals in smoke, involving nitric oxide oxidation
to nitrogen dioxide and subsequent reaction with smoke constituents
such as isoprene, butadiene and acrolein (Pryor, et al., 1983a; Pryor, et
al., 1984; Cueto & Pryor, 1994; Church & Pryor, 1985):

 Page 428
In this scheme, it is postulated that most of the radicals are generated
from nitrogen dioxide in reaction (2). Reactions (3) and (4) are radical
propagation reactions and reactions (5), (6) and (7) are radical
termination reactions. Using model nitric oxide/air/olefin mixtures
and both ESR spin trapping (Pryor, et al., 1984) and Fourier transform
infra-red spectroscopy techniques (Cueto & Pryor, 1994), Pryor and
coworkers have demonstrated that this steady-state radical system is
produced.
The build-up of radicals as the smoke ages, illustrated in Figure 12.14,
is paralleled by the changes in nitric oxide and nitrogen dioxide levels
depicted in Fig. 12.11. It again illustrates the point that smoke
properties change rapidly and that from the smoker's perspective the
relevant smoke properties are those when the smoke is only 1 or 2
seconds old. Furthermore, it must be borne in mind that, as discussed
in Section 7 above, the oxidation of nitric oxide in the isolated gas
phase of smoke (as used by Pryor, et al.) is faster than in whole smoke
(Borland, et al., 1985). Thus the build-up of radicals as whole smoke
ages would be significantly slower than that depicted in Fig. 12.14.

Fig. 12.14
Effect of aging the vapor-phase
of smoke on short-lived
radicals
 (Church & Pryor, 1985.
Section 11 Environmental Tobacco Smoke
Environmental tobacco smoke (ETS) is the material released into the
ambient environment by smoking tobacco products. It is formed as
sidestream and exhaled mainstream smoke diffuse into the
environment. Exhaled mainstream contributes between 15 and 43% of
the particulate matter of ETS and between 1 and 13% of the vapor
phase constituents (Baker & Proctor, 1990). Additional minor sources
of ETS are gases which diffuse out of the cigarette through the paper
during a puff, material released through the butt between puffs and
smoke material released into the environment after being deposited on
surfaces. As the originally concentrated sidestream and exhaled
mainstream smoke diffuse away from the cigarette and smoker, they
become greatly diluted. Various physical and chemical changes occur;
in particular, substances such as nicotine evaporate out of the smoke
particles into the vapor phase, discussed in Section 5, and ETS is
formed. Because of evaporation of material out of ETS particles, the
resultant ETS particles contain a higher percentage of high molecular
weight components than is found in mainstream or undiluted
sidestream particles.
As cigarettes are smoked in a room, the levels of ETS components in
the room rise and then fall due to air circulation, room ventilation and,
to a lesser extent, interactions of the ETS constituents such as
deposition of smoke particles onto surfaces in the room. Some results
of Benner, et al. (1989) in a Teflon bag simulating a sealed room are
illustrated in Fig. 12.10(b). Under these sealed conditions the
decreases in ETS levels after smoking are very slow. The decreases
are much more rapid when there is some ventilation and air
circulation in the room. This is illustrated for two room air exchange
rates for ETS carbon monoxide profiles in Fig. 12.15. Clearly, room
ventilation has a large effect on the decay. For a given set of room
environmental conditions, individual ETS constituents decay at
different rates, as illustrated in Fig. 12.16. (Data in Figs 12.15 and
12.16 were obtained by smoldering two cigarettes in a 30 m3 chamber
(Baker & Proctor, 1990).) The relative rates of decay of some ETS
components are in the order (Baker & Proctor, 1990; Eatough, et al.,
1990):
nicotine > total particles > total volatile organic compounds » NO >
CO2 = CO > NO2
The decay rates of the carbon oxides depend solely on the effects of
air exchange rates, and their levels in ETS remain constant when there
is no air movement

Fig. 12.15
ETS CO profiles for two room conditions (Baker & Proctor, 1990).

 Page 429

Fig. 12.16
ETS build-up and decay profiles
(data from Baker & Proctor, 1990):
two cigarettes smoldered in a 30 m3
chamber, one air change per hour.
(e.g. data in Fig. 12.10 (b)). When an ETS component decays faster
than carbon monoxide there must be some mechanism, chemical or
physical, other than just air movement that depletes its level. ETS
nitric oxide decays faster than carbon monoxide, and nitrogen dioxide
decays at a slower rate due to the conversion of nitric oxide to
nitrogen dioxide by atmospheric oxidation. The faster decay of
particulate mass than carbon monoxide is due to loss of material from
the particles by evaporation to the gas phase and deposition onto room
surfaces. Particulate mass decays faster than total volatile organic
compounds because of some generation of gas-phase volatiles by slow
evaporation out of the ETS particles, as discussed in Section 5.
Finally, the much faster decay of nicotine than ETS particulate mass is
due to it being readily adsorbed by room surfaces and its more rapid
diffusion to those surfaces than aerosol particle diffusion since ETS
nicotine is largely in the gas phase.
In real-life situations, the actual and relative removal rates of ETS
constituents will depend on local environmental factors such as wall
and floor coverings, furnishings, presence or absence of people, air
flow, temperature and relative humidity. Thus, ETS is a constantly
changing mixture due to loss of material as a result of adsorption or
chemical change, changes in gas/ particulate phase equilibrium for
volatile species and desorption of adsorbed species from surfaces.
As sidestream smoke and exhaled mainstream smoke combine to form
ETS, their properties are drastically altered by the great dilution and
other changes occurring. Thus, their properties in ETS are
considerably different from their individual properties at the moment
of their generation. Consequently, the sidestream/mainstream ratios
quoted in Table 12.6 can be misleading if used out of context. The
important question in ETS considerations is not the ratio of
sidestream/mainstream, but rather what is the concentration of the
constituent in the indoor environment and how does it compare to
levels from sources other than ETS. Studies based solely on
observations of fresh sidestream or highly and unrealistically
concentrated ETS should take into account the possible differences
between these smokes and ETS found in real-life situations.
No indoor air environment in real life contains ETS in isolation but
rather in combination with chemicals from a variety of different
sources such as building and cleaning materials, cooking and heating
fuels, consumer product and vehicle exhausts drawn in from outside.
Many of the chemicals associated with ETS will also be present as a
result of such sources. Hence, in order to determine potential
exposures to ETS, it is essential to employ methods that allow a
distinction between substances present as a result of tobacco smoking
and substances present as a result of the other various sources.
Since the late 1980s, investigators have sought a chemical marker for
ETS, i.e. a substance unique to ETS that is representative of the
behavior of ETS as a whole (Baker & Proctor, 1990; Eatough, et al.,
1990; Guerin, 1991; Guerin, et al., 1992). It is probably not possible
to identify one marker because, as discussed above, different
components in ETS behave so differently. Nicotine has commonly
been used as a marker and its presence in air does indicate that
smoking has taken place. However, its behavior in ETS is unique,
both in its rapid surface adsorption and desorption back into the
atmosphere over several hours and days following smoking (Guerin &
Jenkins, 1992; Guerin, et al., 1992). Solanesol is generally considered
to be a good marker for the particulate phase of ETS (e.g. Ogden &
Maiolo, 1989; Tang, et al., 1990). Solanesol is a trisesquiterpenoid
primary alcohol found in tobacco that transfers intact to tobacco
smoke, makes up 2 to 4% by weight of ETS particulate matter and is
essentially unique to ETS in indoor air. Because of its high molecular
weight of 631, it has low volatility and remains in the ETS particulate
phase even at high dilutions (Guerin, et al., 1992). Its concentration in
ETS has been reported to decay rapidly under very intense ultraviolet
radiation (Tang, et al., 1990) but not under realistic, normal
conditions. It is probably the most promising ETS particulate-phase
marker available. Another proposed marker for ETS vapor-phase is

 Page 430
3-ethenylpyridine (Eatough, et al., 1990; Guerin & Jenkins, 1992).
This base is unique to tobacco smoke, is present in the vapor phase of
ETS at measurable concentrations and decays in indoor environments
at rates which are near to those of nonreactive constituents such as
carbon monoxide and dioxide (Eatough, et al., 1987; Eatough, et al.,
1989).
There are many papers available in which measurements are reported
of ETS levels in a whole range of indoor environments, including
offices, buses, trains, aeroplanes, homes, restaurants and bars.
Summaries of these measurements have been tabulated in several
reviews (Repace, 1987; Guerin, 1991; Guerin, et al., 1992; Holcomb,
1993). The most comprehensive summary of data up to 1992 is given
in the review by Guerin, et al. (1992), to which the reader is referred.
Table 12.8 gives a summary of the summarized data, which is based
partly on the weighted arithmetic mean values from studies published
between 1980 and 1993, calculated by Holcomb (1993), together with
additional mean ranges taken from Adlkofer, et al. (1995); Guerin, et
al. (1992), or summarized by Guerin and Jenkins (1992). The average
ranges in Table 12.8 are based on many studies which used different
sampling and analytical methodology which are not always
comparable. The results from one large study conducted in the UK by
Kirk, et al. (1988a; 1988b) have not been included in the averages
because of questions about their analytical methodology (Guerin, et
al., 1992).
These summarized results agree with those of individual studies in
that they show that ETS is a measurable contributor to indoor air
constituents but that the contribution generally overlaps background
concentrations (nonsmoking environments) for those constituents
which have other significant sources. Thus, for example, Proctor and
co-workers (Proctor, 1989; Proctor, et al., 1989a; Proctor, et al.,
1989b) have shown that in smoking-allowed offices, train com-
Table 12.8 Ambient concentrations of ETS constituents in commonly
encountered environments (Guerin & Jenkins, 1992; Guerin, et al., 1992;
Holcomb, 1993; Adlkofer, et al., 1995).
Mean (range) indoors
Outdoors
(typical
Constituent Environment SmokingNonsmoking range)
RSP*(µg/m3) Homes 50 22 (777)
(17212)
Offices/public 68 46 (nd-240)
buildings (121088)
Restaurants 132 90 (nd)
(10685)
Bars, etc. 104 (nd) (nd) 2050
Work areas 36 (nd)
(6519)
Trains 216 185 (64450)
(71325)
Aircraft 57 25 (398)
(3185)
Nicotine (µg/m3) Homes 4 (0.112)0.3 (01)
Offices/Public 6 (nd- 0.3 (0.12)
buildings 70)
Restaurants 6 (037) nd
Bars, etc. 19 (364) 1 (0.42)
Trains 15 4 (0.521)
(0.649)
Aircraft 22 (098) 10 (040)
Carbon monoxide Offices/public 3 (0.19) 3 (0.74)
(ppm) buildings
Restaurants 4 (0.49) nd 14
Bars, etc. 6 (nd) nd
Trains 2 (15) 1 (0.53)
Nitrogen oxides (ppb) Combined 10200 10200 1050
Formaldehyde (µg/m3) Combined 1050 1040
Benzene (µg/m3) Combined 530 525 250
Benzo[a]pyrene Combined 0.32.0 0.21.0 0.10.6
(ng/m3)
N- Combined 10-100
Nitrosodimethylamine
(ng/m3)
* Respirable suspended particulates are those with diameters <5 µm that can
be deposited in the respiratory tract.
ND = not determined or data not available.

 Page 431
partments and betting shops, ETS contributes, on average, 26%, 28%
and 38%, respectively, to the total respirable suspended particulate
levels in the environment.
Respirable suspended particulate (RSP) levels have been reported
from below detection limits to above 1000 µg/m3, but the most
common levels in smoking environments are below about 200 µg/m3.
RSP levels in smoking environments are typically 20 to 40 µg/m3
higher than in the equivalent nonsmoking environment. The data from
the bulk of the studies indicate that the most common ETS nicotine
levels in smoking environments are in the range 2 to 10 µg/m3,
although higher levels are experienced in bars and Smoking sections
of public transport. True background levels of nicotine in
environments where smoking has never occurred would be expected
to be zero. This is often the case, but in many nonsmoking
environments small ambient nicotine concentrations are measured,
due to convection/diffusion from smoking areas and to desorption of
nicotine deposited on surfaces from earlier smoking in the area.
Carbon monoxide levels in smoking environments are generally
similar to those in nonsmoking environments, and the levels in most
environments are dominated by carbon monoxide from sources other
than ETS, including vehicle exhaust, cooking and heating sources.
Similarly for nitrogen oxides, formaldehyde, which is emitted from a
number of consumer products and building materials, and benzene.
Individual studies do, however, differ in their conclusions. Thus, for
example, Wallace, et al. (1987) and Wallace (1990) found excess
concentrations of benzene in smokers' homes compared to
nonsmokers' homes of 3.5 ± 1.8 µg/m3. However, other more recent
studies have found the excess concentrations to be no more than 1.6
µg/m3 in smokers' homes in American and German cities. On the
other hand, Bayer and Black (1987a; 1987b), Proctor, et al. (1989b)
and Proctor (1989) found no difference in benzene levels between
smoking and non smoking offices and train compartments or in
personal exposure of nonsmoking women in different smoking
situations (Proctor, et al., 1991). Furthermore, a personal exposure
study on a nonsmoking cyclist commuting in an urban area (Bevan, et
al., 1991) has reported mean outdoor benzene levels of 17 to 82
µg/m3, dependent on traffic and weather conditions. Levels of 2
µg/m3 were detected in city parkland and 364 µg/m3 at a distance of 2
m behind a stationary car.
These results on benzene demonstrate the uncertainty in individual
studies due to the difficulty of identifying completely ETS-free
environments and of avoiding other sources of the target chemical.
This uncertainty has been addressed when two independent personal
exposure monitoring studies (Adlkofer, et al., 1995; Heavner, et al.,
1995) have been undertaken to determine the percentage of various
volatile organic compounds present in smokers' homes that could be
attributed to ETS. For benzene, 13 to 15% present in the atmosphere
inside the smokers' homes in cities in Germany and USA is
attributable to ETS (Adlkofer, et al., 1995; Heavner, et al., 1995). For
27 other volatile organic compounds investigated in the indoor air of
American homes, there was no statistical difference in concentrations
of 21 of them between smoking and nonsmoking homes, and one
substance (trichloroethylene) was elevated in nonsmoking homes
(Heavner, et al., 1995). For those volatile organic compounds elevated
in smoking homes, the percentages that could be attributed to ETS
are: total volatile organic compounds (5.5%), benzene (13%), styrene
(13%), pyridine (41%), 2-picoline (67%), 3-picoline (90%), 4-picoline
(37%) and 3-ethylpyridine (62%). In these American smokers' homes,
indoor air sources other than ETS were also identified for limonene,
tetrachloroethylene, 1,4-dichlorobenzene and alkylbenzenes (Heavner,
et al., 1995). This type of information is essential in order to put ETS
in real-life environments into proper perspective.
Summary
Tobacco smoke is a continually changing mixture of 4000 or more
components which are present in mainstream and sidestream smoke
and distributed between a particulate and a vapor phase of the smoke
aerosol. This chapter describes the combustion processes occurring
inside the cigarette burning zone together with the mechanisms which
lead to smoke generation, including formation of environmental
tobacco smoke (ETS).
Physically, mainstream smoke is a highly concentrated aerosol whose
particles rapidly undergo coagulation and can absorb or lose volatile
material to the surrounding gas phase by condensation or evaporation.
This results in the physical and chemical nature of smoke changing
within fractions of a second. Sidestream smoke particles are initially
smaller than mainstream particles, and the sidestream plume
accelerates as it rises above the cigarette. As sidestream smoke and
exhaled mainstream smoke enter ambient air to form ETS, they are
greatly diluted and an increasingly larger fraction of the more volatile
con-

 Page 432
stituents, including nicotine, evaporate from the aerosol particles into
the vapor phase.
The concentration of the chemical constituents in smoke changes
within seconds as fresh smoke ages. This together with some artifact
formation of substances in some of the analytical procedures
employed can lead to uncertainties in the relevance to the smoker of
some smoke analytical data. In addition, the concept of 'smoke pH'
has often been over-interpreted in the past, leading to unjustified
conclusions being made on its relevance.
The chemical formation of a variety of representative constituents that
make up smoke is summarized, together with their distribution
between the mainstream and sidestream. Experiments involving the
pyrolysis of possible smoke precursors can give insights into the
reaction pathways occurring in the cigarette. However, unless these
experiments are performed under the dynamic conditions that occur
during tobacco burning, their results can be misleading. This and other
conflicting chemical data are rationalized as much as possible
throughout the chapter. The buildup and decay of ETS constituents in
indoor air are also reviewed, and levels in a variety of environments
summarized. In these environmental studies, care needs to be taken in
attributing the contribution which ETS makes to the total air burden
since many substances in ambient air arise from several sources.
Studies which address this aspect have been emphasized.
The vast majority of information in this chapter refers to cigarette
smoke because cigarettes have been studied in far greater detail than
other smoking articles. Many of these data have been obtained when
the cigarettes were smoked using smoking machines under
standardized conditions. The machine-standard smoking methodology
is discussed, including its relevance to human smoking, and the use is
placed into its proper perspective.
Acknowledgements
I would like to thank the following colleagues for making many
helpful comments on various aspects of this review during its
preparation: Ian Anderson, Peter Bevan, Robin Crellin, Mike Dixon,
George Few, Richard Gammons, Hugh Honeycutt, Geoff Hook, Derek
Irwin and Werner Schneider. I am particularly indebted to Charles
Green and Alan Rodgman for making innumerable suggestions for
improving the accuracy and, hopefully, clarity of the text.
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Chapter 13
Cigars and Cigarillos
Adeler Frederik Wehlburg
ASP Enterprises, Inc
Guayaquil, Ecuador
Introduction
One might wonder why a tobacco monograph would dedicate a
chapter exclusively to cigars and cigarillos. There are quite a few
good reasons for that, as 'black tobacco' in many ways differs from
'blond tobacco'. There are differences in tobacco varieties, tobacco
farming practices as well as in manufacturing, marketing and
consumption.
Cigar smoking in the USA has been experiencing a spectacular boom
since late 1992, causing a serious shortage of black tobaccos
worldwide, and the end of this boom is yet not in sight. Cigar
Aficionado, the magazine for cigar lovers that was launched in the
USA in the autumn of 1992 and which had a major contribution to the
renewal of interest in cigar smoking, explains in its autumn edition of
1996:
'Only four years ago, making or selling cigars may have been one of the
least attractive businesses in the United States. Not only were cigar sales
flat, but the future promised more of the same. The core customers base
consisted primarily of older men. With few new customers venturing into
cigar shops, the industry was literally fading away. Throughout the 1980s
and early 1990s, premium cigar imports the best indicator of cigar sales,
as most handmade cigars are made outside of the United States were
stagnant, hovering around the 100 million mark. Then, in autumn of 1992,
the industry began to change.
 Imports into the USA topped 240 million cigars in 1996, and cigar sales
have continued their remarkable performance. The premium cigar market
more than doubled in the span of 3 years. Not bad for a business that was
ready to be written off only 4 years previously. Today, there aren't nearly
enough cigars to meet demand. Cigar Aficionado estimates that total
industry back orders of premium cigars reached 55 million units in 1996,
approximately double their estimated level at the end of 1995. In part, the
back-order problem exists because the tobacco in cigars takes at least 18 to
24 months to go from seed to a finished cigar. Even manufacturers with the
foresight to increase production in 1994 found that it required more than 2
years to bring in significantly larger supplies of tobacco; no one predicted
the level of demand in 1996, so it will take at least several more years to
reach an equilibrium.
The back-order situation is even more impressive when you consider the
rising prices that have emerged in the cigar market since the dawn of the
boom. In early issues of Cigar Aficionado it was common to see cigars
retailing for under US$2. Today, such an inexpensive cigar is a rare find.
The meat of the market today is up to US$5 per cigar, and it is not
uncommon for a new brand to sell for USS10 per cigar or more.'
What is a Cigar and What Makes it so Different from a Cigarette?
A cigar is a 100% natural product and consists of three parts: the filler,
binder and wrapper. The filler is rolled into a binder leaf, and this so-
called bunch, on its turn, is covered with a wrapper leaf. It is the
combination of a certain filler blend with a specific binder and a
specific wrapper which makes the taste of a cigar unique. Cigars are
made by hand or by machine. The cigar smoke is not to be inhaled. It
is a pure pleasure for mouth and nose and the aftertaste and smoke
satisfaction last considerably longer than in the case of cigarette
smoking. The premium cigar is a luxury product like a good wine, a
cognac or a whiskey is. As a gift, a box of cigars is highly appreciated
by a cigar smoker, whereas you hardly bring a pack of cigarettes as a
birthday present for a cigarette


 Page 441
smoking friend. There are, however, changes to the traditional cigar
concept as the industry continues its search for new products and cost
reductions. In some cigars, mainly the smaller European style
cigarillos, reconstituted tobacco sheet and expanded tobacco are used,
while some cigars are flavored or have a filter or a mouthpiece.
The Filler
Cigar filler tobacco must have taste, a good burn and a certain filling
power. White ashes are always appreciated but not strictly necessary
for a good taste. The leaf does not have to be perfect nor beautiful as it
is torn apart or even shredded to be put inside the cigar.
a
Long Filler
Most of the bigger, high quality cigars (the premium cigars) are made
by hand out of long filler. Long filler consists of a blend of parts of the
tobacco leaf, putting together tobacco of usually not more than three
different filler varieties. The cigar maker has to be very consistent in
his selection of the parts of the several filler leaves he uses. Anyone
who has seen a cigar maker roll a long filler cigar by hand will admit
that cigar making is an art and that these tabaqueros are real artists.
The difference between a long filler cigar and a short filler cigar is a
matter of personal appraisal and therefore somewhat subjective. Long
filler cigars tend to burn more slowly and to give a cooler air flow
than short filler cigars. Some consider these characteristics as an
additional pleasure in cigar smoking.
b
Short Filler
Most short filler cigars are machine made. The tobacco leaf is
shredded like cigarette filler and wrapped in a binder leaf, and this
bunch is subsequently rolled into a wrapper leaf by machine. Here,
one can blend as many different tobaccos as one wishes, thus
spreading the risk of having to change a specific blend because of a
sudden shortage of a certain variety of filler tobacco, as one variety of
filler tobacco contributes less to the final taste of the short filler cigar.
Besides, the different filler tobaccos are mixed in large drums and
often regulated by computers for the consistency of the specific blend.
In other words, this means fewer problems for the manufacturer about
maintaining the filler blend consistency.
The Binder
Any leaf which is too nice to be used as filler and not suitable to be
used as wrapper is used as a binder leaf. Of course, the cigar binder
must have taste and a good burn, because manufacturers will have
certain preferences for their different brands as the binder does
contribute to the taste of the cigar. A binder leaf may have spots and
should be thicker, and thus stronger, than a wrapper leaf, but should
not have holes as it has to seal the filler blend. On the Other hand, the
binder cannot be too thick and should not have pronounced veins as
this would have an adverse effect on the appearance of the final
product because the wrapper is a very thin leaf which has its limits in
hiding these characteristics. Experienced cigar makers can use two
smaller cuts as binder to seal the filler.
The Wrapper
The wrapper is the most beautiful leaf of the cigar as it is the only leaf
exposed to the smoker's eye and, therefore, an important factor for the
manufacturer; it's his business card. This beauty has its price; wrapper
is the most expensive tobacco leaf on the market. Some specific
wrapper varieties are sold for more than US$100 per kg as compared
with US$15 to 20 per kg for exclusive filler tobaccos. The wrapper
leaf has to be perfect because only a perfect leaf is fully usable. It has
to be a thin leaf with a nice stretch and shine, uniform in color,
without spots or holes and with as little (read small) veins as possible.
Obviously, this requires a tremendous effort in production practices
and control from the seed to the packing of the final bale, which
explains its high cost. Cigar wrapper production is a very labor
intensive process; from seedbed to packed bale some 12 000 man-
hours per ha are needed.
However, it is not only the appearance of the wrapper leaf that counts.
The taste is also important. As taste is subjective, a variety of opinions
exist on how much a wrapper leaf contributes to the final taste of a
cigar. Estimations go all the way from some 30 to 80%. Of course, the
impact of the wrapper leaf on the taste of the cigar depends much on
the variety of the cigar wrapper tobacco used. Manufacturers use
dominant wrapper varieties for certain brands, like Cuban, Cameroon
and Sumatra leaf, whereas for other brands they prefer mild or more
neutral varieties like Connecticut, JavaIndonesia, Mexican and
Ecuadorian wrappers. Finally, the smaller the cigar, the stronger the
influence of the wrapper leaf on the taste of the cigar.

 Page 442
Farming Practices of Cigar Tobacco
Cigar tobacco is air-cured, but dark air-cured tobacco does not
necessarily have to be cigar tobacco. Dark air-cured tobaccos are used
in cigars, cigarettes, pipe tobacco blends, chewing tobacco and snuff.
When discussing cigar tobacco varieties and their farming practices,
one has to take into account whether the dark air-cured tobacco is
used for cigar filler or for cigar wrapper, for reasons explained earlier.
The most sophisticated approach is seen in the cigar wrapper
production. Focusing on a discussion of cigar tobacco, it is necessary
to distinguish between cigar filler and cigar wrapper tobaccos when
covering such topics as agronomic practices and varieties (cultivars).
Tobacco is tobacco, however, and in many ways the production
practices for cigar tobaccos are the same as or very similar to other
tobacco types. Therefore, this chapter emphasizes the differences in
farming practices between cigar tobacco and other types of tobacco,
rather than attempts to create a complete manual for the production of
cigar tobacco. Akehurst (1981) discussed the agronomic aspects of
cigar tobacco production.
General Requirements
a
Agronomic
In general, the environmental conditions required for cigar tobacco are
very similar to any other variety of tobacco. Sandy loam soils with
good aeration and drainage and a relatively high organic matter
content are ideal. High organic matter contents of 3 to 4% are no
obstacle for cigar tobacco; on the contrary, it gives the leaf its
chemical structure required for a nice luster and a good stretch.
Filler tobaccos require sufficient direct sunlight for the development
of taste and body. Wrapper tobaccos should not receive too much
direct sunlight as the wrapper leaf has to be fine structured and thin. In
several places in the world, wrapper tobacco is grown under shade
(cheese cloth) precisely for this reason. The cloth results in a
reduction of up to 40% in the sunlight (depending on the density of
the woven cloth) and the entering light is diffused so that it hits the
leaf more uniformly. Furthermore, the micro-climate created inside
the tent is more humid and more uniform (less wind). Lastly, to a
certain extent, the tent keeps (the larger) insects outside. All these
factors help to keep the tobacco plant from stress and, therefore, help
it to grow fast and develop large, thin and uniform leaves without
pronounced veins. However, putting up the tent with poles, wires and
the cloth is a time-and labor-consuming, and therefore cost-increasing,
activity, which should only be considered in areas where sunshine is
too abundant for the growing of good wrapper tobacco.
Rainfall should be moderate and distributed evenly over the growing
season. Again, filler tobacco needs some stress to develop its required
qualities, whereas wrapper should be protected as much as possible
from any kind of stress, which makes additional irrigation often
desirable.
b
Varieties (Cultivars)
There are hundreds of black tobacco varieties that could qualify for
cigar tobacco. Much depends upon where they are grown and under
what circumstances. A tobacco variety could provide leaves suitable
only for filler purposes when grown in a dry environment with a lot of
sun or when grown up in the mountains, but could well provide
excellent wrapper quality leaf when grown in areas with less direct
sunlight, higher temperatures and higher humidity.
Many regions have their own unique black tobaccos. A characteristic
of these tobaccos is that the vast majority of production takes place on
very small farms. It is only in a few areas that, for various reasons,
production has been encouraged to expand and be manufactured into
smoking articles which find markets in other countries. Local
tobaccos (criollos) with their specific characteristics may be used as
such or may be crossed with other varieties with the aim of enhancing
or diminishing certain desired characteristics in a commercial variety
such as taste, leaf form, leaf size, leaf body, leaves per plant, stalk
position of the leaves, stalk strength, blossom development, disease
resistance, burn, etc.
The main characteristics of cigar tobacco are the dark green color in
the field and the fact that this leaf, harvested prematurely compared to
blond tobaccos, gives the desired chemical consistency for air-curing
and fermenting, two processes which are imminent for cigar tobacco.
Leaves harvested mature result in a paper-like, dull and light-
yellowish leaf without any stretch which is unsuitable for cigar
tobacco.
Well known cigar filler tobacco varieties include several Cuban
varieties, such as Vuelta Abajo and Remedio, grown in Cuba or in
other countries; Criollo, Olor and Piloto Cubano from the Dominican
Republic; Parish of Clarendon from Jamaica; Mexican varieties from
the Vera Cruz and the Platon San-

 Page 443
chez area; Jalapa and Estelí varieties from Nicaragua; filler tobaccos
from some areas of Honduras; Cubita from Colombia; Correntino and
Misionero Criollo from Argentina; Amarelinho, Bahia (Mata Fina,
Mata Sul and Mata Norte) and Arapiraca from Brazil; Paraguayan
filler tobaccos grown north-east of Asunción (Flojo and Pasado);
Pennsylvania seed leaf, Wisconsin (binder) and Connecticut broadleaf
(binder) from the USA; Paraguay Bel (PB) from France; Badischer
Geudertheimer from Germany; Beneventano from Italy; Rio Grande
from the Central African Republic; cigar and cheroot tobaccos from
India; Isabela from the Cagayan Valley in the Philippines; and the
Indonesian Vorstenlanden and Besuki varieties, from the Island of
Java.
For wrapper tobacco, on the other hand, there are actually only five
widely used wrapper varieties: the Connecticut varieties, either shade
grown in Connecticut or from other countries like Cuba (shade grown)
and Ecuador (without shade), including some Connecticut broadleaf
varieties; Partido grown in Cuba; Cameroon and the Central African
Republic type wrappers; the several Sumatra varieties grown in
Indonesia itself and the ones grown in Brazil and Mexico, and the
TBN variety developed in Indonesia East Java (shade grown).
Production Practices
Production practices of black tobaccos are in general much less
sophisticated and less mechanized than is the case in production of
blond tobaccos such as Virginia flue-cured and burley. There are
several reasons for this situation. First, tobacco farmers who grow
black tobaccos are relatively poor Third World farmers who have at
the most a few hectares at their disposal which they inter-crop with
other household crops. They have no money for nor access to modern
technology. Inputs are most often pre-financed by the future buyers of
the tobacco crop. Second, labor in these Third World countries is
relatively cheap and often outsmarts a complicated machine for which
no knowledge of maintenance or spare parts is available. Third, in the
case of wrapper growing, it is often impossible to mechanize certain
handling procedures as the leaf damage would be unacceptably high.
Fourth, most new technologies, pesticide and fertilizer product
development, research on disease resistance and science of genetics in
general are based upon and focused on blond tobaccos such as
Virginia flue-cured and burley since these tobacco varieties are, by far,
the most frequently grown worldwide; whereas little research has
been done on dark air-curing or the fermenting process. Many
improvements such as harvesting machines and bulk curing, which
work perfectly well for cigarette filler tobaccos, are of little value for
black tobacco production.
Cigar wrapper production practices are different from the ones for
cigar filler as the wrapper leaf has to be perfect, thus investment in
research and development and new technologies are worthwhile
against the background of the higher prices that a good wrapper leaf
demands. Obviously, investment per hectare is much higher in the
case of wrapper growing because of the attention paid to the crop in
the field and during curing, fermenting and sorting. The voluminous
wrapper production worldwide is done on an estate basis where
management is in the hands of the owners or managers. Small farmers
usually do not have the means for, nor the understanding of, the
detailed care taking of these fine leaves and will obtain not more than
some 10 to 15% of good wrapper quality out of their crop, which is by
a long way insufficient to cover the cost.
In the following sections, the differences between farming practices
for black and blond tobacco will he highlighted, distinguishing cigar
wrapper from cigar filler varieties where necessary.
a
Seedbeds
The majority of the seedbeds for black tobaccos are still natural soil
seedbeds, often covered with rice husk or any other natural material
for a better germination (maintaining humidity) and weed control. For
wrapper growing, trays are used and even some float systems have
been put in use (Mexico). Soil disinfecting, fertilization and pesticide
applications are the same as for other tobaccos. As the soil medium
for the trays, ready-for-use peat moss may be used, or local materials
such as fine humus mixed with rice husk or coconut dust mixed with
some sand. Only recently has clipping (or pruning) become a custom
in cigar tobacco seedbeds. The clipping of the seedlings is done to
obtain stronger seedlings and more uniformity in the seedbeds. It
strengthens the stem and prevents so called spindlings (tall seedlings
with thin stems which easily fall over once planted in the field). Also,
clipping slows down the growth of the seedlings and, therefore, is
practised when weather circumstances prohibit transplanting. Under
tropical conditions, seedlings should be ready for transplanting in
some 30 to 40 days.

 Page 444
b
Cultural Practices
Fertilization
Obviously, the fertilization of a tobacco crop depends entirely on the
specific variety, its intended use and local circumstances including
soil type, nutrients available for the plant in the soil and the rainfall.
Approximate quantities of nitrogen, phosphorus and potassium used
in a black tobacco crop would be: 180 kg/ha N, 120 kg/ha P2O5 and
210 kg/ha K2O, related to a harvest of some 1500 kg/ha. It should not
be assumed; however, that nutrients added in fertilizer should be in
the same proportion as they are thought to be removed by the plant,
because the type and amount of fertilizer required will vary with local
soil conditions and will be affected by what is in the soil already and
other factors.
Most of the fertilizers used in cigar tobacco production are the same
as, or very similar to, the ones used in blond tobacco production, but
the rates are different as we are dealing with tobaccos in which uptake
of nitrogen in the field is much higher. Perhaps the greatest difference
between cigar tobacco and blond tobacco is that nitrogen uptake
should continue up to the time of harvesting. As wrapper tobacco is
considered a high value crop, modern high quality fertilizers with a
relatively high cost are used in order to obtain the specific
characteristics which a good wrapper leaf should have: good body
without being too thick, stretch, a nice luster or shine and a good
combustibility. Typical fertilizers used for cigar wrapper production
are organic meals such as soybean oil meal, cottonseed meal and
castor pomace that are the slow release nitrogen fertilizers to be
incorporated before transplanting. Frequently, triple superphosphate is
also incorporated into the soil before transplanting. Diammonium
phosphate (DAP), sulfate of potash, nitrate of potash and carbonate of
potash are the most popular for side dressing. Filler tobaccos can and
have to do without these rather expensive fertilizers, therefore, a wide
variety of locally made sources of N, P and K are commonly applied
in this type of crop. In general, cigar wrapper tobacco will require
more of every nutrient than cigar binder varieties, and binder will
need more than cigar filler varieties, except perhaps for nitrogen. No
foliar fertilizer applications are used in cigar tobacco production.
Transplanting
Transplanting of the seedlings is done manually or with transplanters
and normally set in a flat field without pre-ridging. In wrapper
production, the space between the rows is often alternated, one narrow
row followed by a wider row. This is to facilitate the harvesting which
is done only in the wider rows (less leaf break-age), reaping two rows
at the same time. Spacing of the plants in the field varies widely in
black tobacco production and depends much on the specific variety
used and the intended use of the final product. Typical wrapper plant
population would be some 22 000 to 25 000 plants per ha, whereas
filler varieties would range from some 16 000 to 30 000 plants per ha.
Pesticides
In cigar wrapper tobacco, it is extremely important to prevent insect
damage and disease, rather than cure them. Wrapper leaf is bought
after the careful examination of every leaf from the samples (hands of
tobacco leaf) taken out of the bale at random. The sales value of a
wrapper leaf declines rapidly when a tiny hole or spot is observed. On
the other hand, pesticide residues play an important role. The grower
has to be very careful in his choice of products and the concentration
and quantities he applies in the field. He has to make sure that the
coverage of the applied product(s) is optimum. With modern
technology at hand, only the highest quality pesticides registered for
tobacco cultivation are used in wrapper production.
It is, therefore, essential to find the balance between a well protected
leaf and low pesticide residues in the final product, together with as
little damage as possible to the environment. This balance has
everything to do with the local environment and the pressure of
infection encountered in the fields. In wrapper production some
pesticides are applied every 7 to 10 days from transplanting until
harvesting is completed. In the case of a sudden insect or fungus
outbreak, additional and/or post-emergence pesticides are used. In
wrapper production, additional, more expensive pesticides are used
compared to filler growing where diseases should be controlled to a
certain extent, but some spots, holes or somewhat deformed leaves do
not influence the usability of the filler leaf and, therefore, its sale
price.
Diseases encountered in black tobacco crops which can cause serious
economic losses are damping off (Pythium spp.) seedbeds; sore shin
(Rhizoctonia solani) during the first 2 weeks after transplanting; frog-
eye (Cercospora nicotianae) a serious problem in wrapper production;
blue mold (Peronospora tabacina); black shank (Phytophthora
parasitica); fusarium wilt (Fusarium oxysporum); hollow stalk
(Erwina caratovora)

 Page 445
mainly on topped tobacco; dead blossom leaf spot (Sclerotinia
sclerotiorum and Botrytis cinerea) mainly when tobacco is irrigated
overhead; root-knot nematode (Meloidogyne spp.) especially in sandy
soils with high pH and when insufficient crop rotation is carried out;
tobacco mosaic virus (TMV) resistant varieties are frequently used;
leaf curl caused by the whitefly (Bemisia tabaci); broomrape
(Orobanche spp.) a serious problem in Cuba; and finally weather
fleck.
Main insect pests in cigar tobacco are cut and bud worms, aphids
(Myzus persicae) and whiteflies (Bemisia tabaci) the latter causing
the leaf curl virus. Finally, snails can, overnight, destroy a day's work
of transplanting.
Field Maintenance and Topping
To avoid introducing diseases into the tobacco via the weeds, cleaning
up is a common practice, carried out simultaneously with ridging up
(two or three times). In wrapper production weeding is especially
important as many fungi are present in the weeds. Fields are cultivated
by hand or machine or by a combination of both. Also, animal traction
is still widely used in dark tobacco cultivation. While ridging up,
some two to four leaves are taken and thrown away in two or three
rounds (hygienic clean up), as these leaves so close to the ground are
often affected by and pass on all kinds of diseases, as well as often
being damaged by cultivation practices. Therefore, the first priming
actually starts with leaf number 5 and up.
Filler tobacco is topped when the bud reaches a few centimeters above
the small young leaves, which under tropical conditions is the case
around 2 to 3 weeks before harvest, thus leaving from 12 to 18 leaves
on the plant. The number of leaves remaining depends on the fertility
of the soil, variety grown and vigor of the individual plant. A topped
tobacco plant will transport all its energy towards the remaining
leaves, as the plant's first priority, to flower and to produce seed in
order to multiply, has been taken away. Thus, topping will make the
remaining leaves larger and heavier and it will give the filler leaves a
fuller taste and a better burn. In wrapper production one does not top
the plant, as this would make the leaf too thick.
Wrapper tobacco plants grown in the shade must be supported by a
string tied around the stalk and fixed onto iron wires above the
tobacco, as the plants under shade grow so fast that they cannot
support themselves at a later stage. This is very labor-intensive as the
strings must be twisted every week around the ever growing stalks.
c
Harvesting
Cigar leaf is harvested green for reasons explained earlier in this
chapter. As an Example, cigar tobacco which is ready for the second
priming is shown in Fig. 13.1. There are many indications which
people take into account to define the moment when cigar tobacco
should be harvested. One is counting the days; in general, and under
tropical circumstances, the first leaves should be ready for harvest
around 45 to 50 days after transplanting (wrapper production) or some
70 days after transplanting for stalk-cut filler varieties. A different
indication is the leaf color: when the color on the edges of the leaf
changes from deep green to lighter green. Another indication is the
'snapping and cleancut' theory: when the leaf is primed it should snap
and give a clean cut without threads. Then the development of the leaf
is important; coming closer to its
 Fig. 13.1
Harvesting cigar wrapper tobacco: second priming.

 Page 446
harvesting point, the leaf smoothens out and becomes less wrinkled.
Obviously, all indications mentioned are often taken into account to
decide when harvesting should start. Cigar filler tobacco is harvested
by leaf, by stalk and often in a combination of both. Stalk-cut filler is
harvested when the middle leaves are ripe, as indicated by a lighter
shade of green. At this stage, the lower leaves of the plant will tend to
be more or less over-ripe and the top leaves will show varying stages
of maturity. Cigar wrapper is harvested leaf by leaf (primed).
Harvesting Cigar Filler Tobacco
The stalks of the filler tobacco are cut near the ground and the plants
are laid on the ground to wilt before they are handled, so the leaves
are not broken off the plants and lost. Five or six plants are speared on
a 4 foot lath by means of a removable metal spear which is placed on
one end of the lath, and the plants are then placed on a specially built
rack for hauling the tobacco to the curing barn. There are many
different ways, depending strongly upon local traditions, on how the
stalks are harvested (one cut or different cuts), how they are attached
to a lath and how they are transported to a barn. Sometimes, the stalks
are cut in several smaller pieces with some three leaves per piece of
stalk. Often, the first few leaves of a filler crop are primed and the rest
of the plant is stalk cut. Primed filler leaf is sewn, twisted or strung
leaf by leaf onto a pole. It is a common practice to leave the filler
tobacco attached to the sticks, to wilt in the field before taking it into a
shed for the curing process. Regardless of the harvesting method in
filler tobacco, one normally distinguishes only three to four leaf
positions: lower leaves, the lower and higher center leaves and the top
leaves. Stalk-cut tobacco, attached to a lath in the field, is brought to
the barn as such and placed on the tiers.
Harvesting Cigar Wrapper Tobacco
Wrapper is harvested leaf by leaf, two or three leaves at a time, with
some 5 to 7 days in between the primings. Ideally, cigar wrapper
tobacco would be harvested one leaf at the time, but this is logistically
and economically hardly feasible. Considering a plant with an average
of 25 useful wrapper leaves, some 10 rounds (primings) are necessary
to complete the harvest. If four leaves harvested successively are
considered as one priming, one obtains on average five to six primings
of good wrapper leaf. Obviously, tobacco leaves from higher primings
are bigger and thicker than the lower leaves. Top leaves are the
thickest but become smaller than the middle primings. The first
priming is normally somewhat greener than the higher primings as it
has received less sunshine. In wrapper harvesting, it is of vital
importance to have sufficient personnel to harvest continuously,
coming back to the same lot after some 5 to 7 days for the next
priming. This requires quite a bit of logistic thinking.
Some wrapper producers in the state of Connecticut, USA, use plastic
sheets laid out in the rows on which the harvested leaves are placed.
After the entire row has been harvested on both sides, the sheet is
taken in by one man sitting at one end of the row, simply by pulling
and rolling it up, and the leaves are placed in plastic baskets by two
other men. The baskets are covered with cloth and transported to the
curing barn. Others use only canastas (plastic or woven bamboo
baskets) and others place the harvested leaves directly into a trailer,
either laid flat or in a vertical position. Wrapper tobacco should be
taken out of the sun and brought to the curing barn as quickly as
possible in order to avoid damage to the delicate leaf. Harvesting and
transporting wrapper leaf will cause a lot of damage if not done with
proper care.
Depending on the variety of cigar tobacco, its plant population and
whether the tobacco is topped or not, approximate amounts of kilos
per hectare vary from some 700 to 1200 for cigar filler varieties and
from 900 to 1500 for cigar wrapper varieties.
d
Curing
Natural Curing
As natural circumstances are rarely ideal during the entire curing
process, heat is used to prevent rotting and to help complete the
chemical processes required to obtain the desired characteristics of the
cigar leaf. Charcoal, wood and gas, and often a combination of them,
are used as heat sources. The important point is to keep the leaf alive
without letting it rot and to protect it from rain and sunlight (sunlight
discolors the wrapper leaf). Therefore a barn or a shed is a necessity.
Under these circumstances, the leaf respiration will continue and
carbohydrates and proteins will be degraded, leaving the leaf almost
without sugars, resulting in an alkaline reaction on smoking it.
Filler tobacco is often cured in simple open sheds, whereas wrapper
tobacco is cured in large and rather expensive closed barns, normally
made out of wood with a zinc roof, with ventilation provided by
means of windows and an open roof construction. A wrapper

 Page 447
barn of 20 m wide by 100 m long is an expensive asset and is
occupied during some 4 to 6 weeks by tobacco leaf of one priming
only from some 10 ha at a time. One can imagine the importance of
having more than one crop per year to bring down overhead costs by
filling up these expensive barns as often as possible during a growing
season. Cigar leaf needs to be in the barn for at least 25 to 45 days,
depending upon weather circumstances, tobacco variety and on the
specific priming; lower primings need less time to complete the curing
process than the higher primings.
When the harvested leaves arrive at the barn from the fields, they are
sewn face to face and back to back by hand or by machine onto
wooden sticks or bamboo poles (Fig. 13.2). Depending upon the width
of the sections (bents) of the barn, the poles vary from 1 m to as much
as 4 m, carrying 44 to 160 leaves per pole. Smaller leaves and higher
primings can support more leaves per pole than the larger leaves and
lower primings. Consequently, the poles are placed onto the
 Fig. 13.2
Sewing harvested cigar wrapping
leaves on a pole by hand.
tiers at certain distances (Fig. 13.3), depending on the weather
conditions, on the specific priming and on the size of the leaves; poles
containing smaller leaves can be placed closer to each other, and
higher primings tend to have fewer problems with rotting than lower
primings and can therefore be placed closer to each other as well.

Fig. 13.3
Placing the poles on tiers inside a curing barn.
The curing process can differ significantly depending on the variety of
tobacco, its intended use, outside air humidity (rainfall and sunshine
hours), the design of the curing shed or barn and the types of heat
source. It is impossible to explain anything about air-curing cigar
tobacco other than in very broad terms. The same tobacco with the
same intended use but grown only some 50 km away could require
very different curing methods because of different climatic conditions.
The main goal of curing cigar tobacco leaf is to let the leaf die slowly
so that it obtains the desired color, taste and burn. The leaf color is
mainly defined during the curing process, whereas taste and burn
potential are developed more during the fermenting and aging
processes. Curing and fermenting are two closely related and
indispensable processes which cigar tobacco has to undergo. Cured
dark air-cured tobacco has a raw and somewhat irritating taste and
therefore needs to be fermented afterwards.
If the leaf dies too quickly, it will remain green and have poor burning
capacity with a nasty harsh taste. If it dies too slowly, it will either rot
or become too yellow and it will lose the desired characteristics of
luster and stretch.
The dying process of the tobacco leaf starts as soon as the leaf is
harvested in the field. It is, therefore, important to have the tobacco
hung inside the barn as soon as possible to be able to control the dying
process

 Page 448
of the leaf. This is somewhat more important for wrapper than for the
less vulnerable filler tobacco. Once the barn or shed is filled with
tobacco leaf, the curing process can be controlled by the decision
making of the grower. Some allow the leaves to wilt a few days,
others decide to heat up almost immediately. The first firing is
continuous and could last from 3 to 6 days as temperatures will
gradually go up, pushing the relative air humidity down to the desired
level. Once the lamina are dried, the heat is shut off and humidity is
allowed into the barn to moisten the dried leaves. Some growers will
have a second firing of another few days, others will only fire up
again in the case of high humidity or when any rotting of leaves is
observed.
The curing methods really boil down to playing around with the air
humidity inside the barn. The leaf dries out when the relative humidity
becomes 80% and less, and the leaf will be kept alive with a relative
air humidity of more than 80%. With regard to the entire curing
process, it is a rule of thumb to keep the inside relative air humidity
some 75% of the time below 80% and some 25% of the time above
80%. In this way, the leaf will cure slowly. Generally, one seeks to
have the inside temperature maintained at some 2° to 4°C (5° to 10°F)
higher than the outside temperature and the percent relative humidity
some 10 points lower than the value for the inside temperature in
degrees Fahrenheit. For example, if the inside temperature is 90°F
(32°C), the relative humidity should be approximately 80%.
Curing is characterized by two major interrelated biological events,
regardless of the specific procedures employed. One is the
dehydration which removes 80 to 90% of the original green weight as
water. The other is a series of complex biochemical changes which, in
conjunction with controlled water loss, result in characteristic leaf
colors indicative of desirable chemical and physical leaf properties
which make a particular tobacco suitable for use in a specific
consumer product. The complex physical and chemical changes
involved in air curing begin when the tobacco is harvested and end
when the mid-ribs of the leaves are dried out (Fig. 13.4).
When the curing process is completed according to the subjective
opinion of the grower, the poles are taken down, leaves of one pole
are bundled and tied into several hands and the cured tobacco is put in
boxes for transport to the fermenting barn. The taking down has to be
done in the early morning (often people start at 2 o'clock in the
morning) as humidity is required to soften the delicate cured leaves
for handling and in order for the appropriate reactions to proceed in
the

Fig. 13.4
Curing cigar wrapping tobacco.
Left: first firing of fresh tobacco
by means of gas burners. Right:
almost fully cured leaves; some
charcoal is lit for the prevention
of molds and rot.
fermenting piles: dry tobacco will not enhance the desired enzyme
changes during fermenting. Small filler tobacco growers often create
simple and small fermenting piles inside the same open style curing
barn as they have no means for special fermenting barns.
Serious losses can be caused during the curing process, when weather
conditions are unfavorable or when the tobacco is too wet and no
immediate action is taken. Pole sweat or house burn are the most
commonly used terms for the rotting of tobacco in the curing barns.
Botrytis cinerea and Rhizopus arrhizus are casual agents of web rot,
which destroys all tissue between the main veins. Stalk-cured tobacco
can suffer from a rot of the stalk which may extend to the leaves. The
principal agent is Sclerotinia sclerotiorum, but the bacterium Erwina
caratovora can also be involved. These stalk-rot infections can be
acquired in the field and continue to flourish under suitable conditions
in the curing shed. All these rots betray their presence by smell.
Candela Wrapper
Cigars with green wrappers were quite popular during the 1960s and
the 1970s, and still today attract a certain category of smokers.
Candela wrapper is harvested green, even somewhat more unripe than
natural wrapper. Although perhaps many cigar wrapper varieties
would qualify for the making of candela wrapper, Connecticut
varieties are mostly used. Apart from the general requirements of a
wrapper leaf mentioned earlier, a good candela wrapper leaf should be
uniformly light green in color and should have no pro-

 Page 449
nounced nor white veins and certainly no stains or spots. The barn has
to be filled as quickly as possible and firing should start immediately.
The idea is to kill the harvested leaf as quickly as possible in order to
maintain the green color, and therefore the barn is heated up quickly,
strongly and continuously, reaching temperatures as high as 71°C at
the end of the drying period.
It takes some 50 to 60 hours for the green leaf to dry out completely;
mid-ribs should be brittle and break when bent. Inside temperatures
should gradually and always go up until the end of the drying process
in order to avoid condensation on the leaves causing stains and spots.
Drying candela wrapper is a very meticulous process, and not without
danger, as very high temperatures are obtained by means of gas
burners inside wooden barns. Not least, it is also exhausting for the
workers who will have to be present inside the barn at all times to
carefully watch and control the process. Candela wrapper is not
fermented and is sorted in just a few grades.
Obviously, when sale prices are the same, candela wrapper production
has several advantages to the tobacco grower compared to other
wrapper. First, return on investment is more attractive as candela
leaves can be packed ready for use only a week after harvest. Second,
overhead costs are lower as fewer curing barns per ha are needed and
no fermenting space is required. Third, no fermenting and no
extensive sorting means less handling of the leaf and thus less
breakage.
e
Fermentation
The fermentation of cigar tobacco is a natural continuation of the
ripening and curing processes where carbohydrates and proteins are
further degraded and where aromatic characteristics are developed, or
anyway become more pronounced. The nicotine content will diminish
somewhat during the fermenting process, although not by much. As
long as the moisture content of the tobacco is sufficient (18 to 24%),
the fermenting process is mainly the work of enzymes. At much
higher moisture levels in the tobacco, and in the absence of sufficient
oxygen, certain anaerobic bacteria (Clostridium spp.) come into action
which could promote not only damage to the cellular content but also
to the cell structure, with the inevitable result of weakening the
strength of the tobacco leaf or, worse, decayed leaf. Black rot
(Aspergillus niger) becomes a threat as well, when tobacco is
fermented at too high temperatures, darkening the leaves and causing
varying degrees of disintegration.
Bulk Fermenting
Moisture content of the leaf, outside air temperature and humidity,
together with a certain pressure are the main requirements for a well
developed fermenting process, apart from a nice bodied, well-cured
tobacco leaf. Depending upon the tobacco variety, the priming and
whether the tobacco leaf is still fresh, not yet fermented or already
somewhat fermented, the rule of thumb is that leaf moisture content
should be around 20%, minimum outside temperatures around 21°C
with an air humidity of some 75 to 80% and a pressure of some 200 to
600 kg/m2.
The moisture content of the tobacco leaf obviously has a direct
relationship with the outside air humidity: too low an air humidity
level will dry out the tobacco which will stop the fermenting process,
whereas too high an air humidity level will cause wet tobacco which
could promote mold and rot. Hence, strong fluctuations between
outside air temperatures and the moisture content are unfavorable for
the development of a good, steady and complete fermenting process.
One tends to put less tobacco in a pile for thin leaves as cutters than
for tips; the same for tobaccos on the wet side. Therefore an average
bulk consists of some 2000 kg of wrapper leaf and up to 6000 kg for
filler varieties. Although the fermenting process is a self-generating
heat process, a certain minimum initial temperature is needed for a
good start to fermentation. Enzymes will only be moderately active
below around 16°C. Furthermore, the fermenting process is an aerobic
process and therefore oxygen, and thus air, is needed. During all the
transformations within the leaf, oxygen is taken up and carbon dioxide
released, meaning that for a good course of events, a frequent
refreshing of air is needed.
The traditional and most simple method of fermenting is putting
tobacco in piles or bulks within a closed environment (no rain or
sunshine). Bulks have the advantage of an easy heat accumulation and
a better conservation of moisture. Bulks for wrapper tobacco are
normally established by putting the hands of tobacco manually one by
one in straight rows and in several layers (e.g. 2 m wide by 5 m long
and some 1 1\2m high). Filler tobacco bought by exporters is often
preconditioned mechanically in steamdrums and bulked as loose
leaves on pallets With the aid of removable containers walls.
Under the right conditions, the temperature inside

 Page 450
the bulk, when the fermenting begins, will rise quite quickly, and
when the allowed maximum temperature has been reached, the
fermenting pile will be opened up and turned. Inside temperatures are
measured by means of a thermometer on a string inside a plastic tube
placed halfway in the bulk. Specific maximum inside temperatures
vary a lot according to the variety of the tobacco and the specific
priming: heavy fillers can go up to 60 to 65°C, whereas delicate
wrapper leaves cannot support more than some 42°C in order not to
start 'burning' (convert into black tobacco as a consequence of over-
fermenting). Once the specific maximum temperature is reached, piles
are opened up and reconstructed: 'hot inside tobacco' will be placed
outside, 'cold outside tobacco' will be placed inside and tobacco hands
which lay on top of the pile will now be placed first (on the bottom of
the new pile), and vice versa. It is important to shake the hands of
tobacco leaf gently so that flat and pressed leaves within the hands
will loosen up and fresh air (oxygen) will come in between the
tobacco, before the hands are piled up into the next bulk. When the
bulk does not reach the desired maximum temperature within some 9
days, it should be opened up and turned anyway, because the
fermenting process apparently did not begin because the tobacco was
bulked too dry, or the tobacco dried out because of the outside air
temperature and humidity or the bulk pressure was not sufficient. The
process of turning piles can be repeated some four to seven times as
long as inside temperatures in the new bulk keep on rising: hence, the
entire fermenting process can take up to some 70 days.
The fermenting process is complete when the inside temperature of
the bulk rises much more slowly, reaches much lower levels within
the 9 days and ultimately remains almost constant. At this point, the
fermenting matter has been depleted. Apart from this rather objective
way of controlling and judging the fermenting process, there is, of
course, the important subjective judgment of the tobacco man, looking
at the fermenting leaf at least every time a pile is turned (Fig. 13.5)
and very often even in between two turnings, taking out samples of
leaf from each pile, touching, comparing and smoking them. When
the desired color is set and the smoke characteristics of the leaf are
satisfactory, one often decides to pack the tobacco for sorting without
taking great account of the number of days of fermenting.
It is essential to pack only well-fermented tobacco leaf, otherwise, by
taking up some moisture from the air, the tobacco leaf could provoke
anaerobic fermentation (without air and thus oxygen) within the bale,
with the result of loss of leaf strength.

Fig. 13.5
Turning a fermenting pile of cigar
wrapper tobacco by hand.
Carton Box Sweating
Obviously, the bulk method requires quite some space, time and labor.
Also, it is not a very uniform process as the conditions of each pile
can differ. Therefore, people have always searched for better and more
effective methods for the fermenting process and obviously thought of
smaller units in conditioned rooms for better control and more
uniformity. Carton box sweating was developed for wrapper tobacco
on this basis, and it consists of putting the cured hands of tobacco in
smaller entities (some 60 kg) into carton boxes or wooden cases,
placing them in a completely conditional and preheated room. The
rooms have a starting temperature of some 40°C and are gradually
heated up, maintaining an air humidity level of some 80%. Under
these conditions, the tobacco heats up quickly and rapidly reaches the
same temperature as the forced outside temperature and even more:
some 2 to 5°C above the air temperature. Fermenting within the box is
more uniform as there are less marked temperature and humidity
differences between the 'cold outside tobacco' and 'warm inside
tobacco'. Tobacco temperature is measured by thermometers put into
one box of every lot which goes into the hot room. The boxes are not
opened or moved as long as the tobacco temperatures develop
according to the established schemes. If the temperatures do rise too
much (or too little), that particular lot is taken out, reconditioned,
repacked and placed back into the hot room.
With this forced fermenting process in conditioned rooms, one can
obtain well fermented tobacco within some 2 to 3 weeks instead of 5
to 8 weeks as in the bulk method. Whether the two described
fermenting systems will give fully comparable results is a subjective
matter. Some people swear the results of forced fer-

 Page 451
menting are the same as or even better than the traditional way of
bulking the tobacco. Others claim that tobacco fermented in hot rooms
is not fully fermented and will keep on 'working' and thus changing
color. It is obvious, however, that economics prohibits the timeand
labor-consuming method of bulk fermenting outside developing
countries. In Connecticut, USA, forced fermenting has been put
successfully into practice for more than half a century.
Harvesting, curing, fermenting and aging of the tobacco leaf is a
natural and continuous process of the dark type tobacco, which must
be accomplished and understood in this sequence of events. Some use
the term drying instead of curing and curing instead of fermenting,
which, obviously, is somewhat confusing. Anyway, aging is the
slower continuation of the fermenting process within a pile, bale or
even the cigar itself, which is often called the finishing touch of the
tobacco as it enhances taste and improves burn. Under favorable
conditions, one could keep well fermented tobacco aging for decades
without having to be afraid of loss of quality. On the contrary, well
kept, fully aged tobacco is a highly appreciated merchandise but
unfortunately a rare find nowadays, as also in the dark air-cured
tobacco production, economics dictates the game.
Finally, a poor crop cannot improve during curing and fermenting,
whereas a good crop can suffer from poor curing and/or poor
fermenting.
Sorting
After fermenting, dark air cured tobacco normally has to be sorted in
one way or the other to be accepted by the ever demanding client.
Filler tobacco is mostly sorted on conveyer belts in just a few grades
based on the former stalk position of the leaves which determine their
characteristics. Depending very much on local traditions, filler
tobacco is packed in bales of some 50 to 70 kg and the packing
material used is often local natural material like jute, bamboo, palm
leaves, etc.
Wrapper tobacco, on the other hand, is sorted manually, leaf by leaf,
and is classified sometimes into as many as 1500 final grades.
Important characteristics are the color, quality and length of the leaf.
Quality depends on the uniformity of the color within the leaf and
whether the leaf has dark tips, the pronunciation and color of the veins
of the leaf, the body of the leaf and its stretch and whether the leaf has
spots or holes. A wrapper tobacco leaf easily goes through more than
a hundred pairs of hands from harvesting to the finished product.
Before the tobacco leaf is handled it has to be conditioned with water
by means of vaporizers, depending again on the variety of tobacco. A
general leaf moisture content of some 18 to 22% is required to avoid
breakage. On the other hand, over-conditioned tobacco leaf will easily
tear apart. After the bundles have been moistened, the leaves are
untied and opened up for the selection on color and grade. Once
sorted on these bases, the leaves are measured (sized) and bundled
again in 'commercial' hands of around 30 to 40 leaves each. These
hands are to be dried again in a hot room with electrical or gas heating
systems and packed into bales of some 50 to 60 kg. Wrapper leaf is
normally packed at a moisture content of some 15 to 16%. Tobacco
packed too wet will promote the growth of fungi and rot. Leaf packed
too dry will cause a lot of breakage as the hands of wrapper leaf are
pressed before being packed into a bale. The wrapper leaf is put neatly
into a more or less square shape of about 1 square meter; some protect
the expensive leaves by placing carton pads at the edges of a bale
before it is wrapped and sewn up with fine, natural fiber cloth.
Bales of tobacco will remain for several months in the warehouse of
the producer/exporter to age the tobacco leaf within the bale to some
extent; the bales are turned from top to bottom twice a month. Most
manufacturers will continue this aging process for their account once
they have the tobacco in their possession. Some even open up the bale
and re-ferment and re-sort the purchased tobacco to their liking. In
general, aging of the tobacco is more vital for the larger, long filler
premium cigars than it is for the smaller, short filler cigarillos.
Cigar manufacturers will come to the country of origin for the careful
and time-consuming revision of an offer of a few hundred bales of
cigar tobacco. In the case of wrapper tobacco, machine made cigar
manufacturers need to be certain that one bale of tobacco leaf (some
350 hands of 35 leaves each) consists of exactly the same grade
(color, quality and size). They cannot afford to have people select the
manufactured cigars on basic colors before they go to their final
destiny: the cigar box.
References
Akehurst, B.C. (1981) Tobacco, 2nd edn. Longman, London and New
York.

 Page 452

Chapter 14
Smokeless Tobacco
Inger Wahlberg
Swedish Match Sverige AB
Stockholm, Sweden
Tommy Ringberger
Swedish Match Sverige AB
Stockholm, Sweden
Introduction
Seen in a historic perspective, smokeless tobacco is a modern
expression for a traditional product category. This category includes
tobacco products that are sniffed in the nose, sucked or chewed in the
mouth. Smokeless tobacco is hence not burned to produce smoke for
inhalation (therefore smokeless). It is enjoyed in direct contact with
oral or nasal membranes, where it is allowed to deliver its flavors.
The scientific literature on smokeless tobacco products is sparse
compared to that on smoking products. In-depth chemical studies have
not been published, and manufacturing methods and ingredients are
generally well protected secrets of the manufacturers. It has therefore
not been possible to give a comprehensive survey of the field in the
present chapter. Our intention has been rather to give a brief
description of some of the many types of smokeless tobacco products
encountered around the world today. The tobaccos most commonly
used in such products are presented and so are some of the
manufacturing methods described. Chemical characteristics of some
smokeless tobacco products are provided.
Product Types
Smokeless tobacco products are traditionally classified as snuff or
chewing tobacco. This classification was originally based on how the
products are used. Snuff, which was generally sniffed in the nose, was
by necessity a more disintegrated product, i.e. ground or fine cut, than
were products for oral use. The latter were in general classified as
chewing tobacco. This division is still valid in countries under
previous British, French or German linguistic influence. In the United
States and in the Scandinavian countries, however, the term snuff (or
snus) has, for centuries, also been used for oral smokeless tobacco
products which have a finer texture.
For legislative purposes, some countries have a legal or fiscal
definition of smokeless tobacco products which does not always
coincide with the traditional terminology.
Because of a lack of universal definitions, statistics on smokeless
tobacco from different countries are often confusing and
misinterpreted. The consumption statistics rarely tell the difference
between oral and nasal snuff. The word snuff itself is interpreted as a
nasal product in most languages.
Hence, the traditional classification into chewing tobacco or snuff
does not express how the products are used today. Nor does it reveal
the visual appearance of such products. Chewing tobacco may or may
not be chewed (in fact most chewing tobacco products are kept in the
mouth and not chewed). The appearance of chewing tobacco can vary
from finely grated tobacco particles to blocks of compressed tobacco
or thick strands of spun tobacco. Snuff may be used as a fine tobacco
powder in the nose, but is today more commonly used as an oral
tobacco product. Snuff is, however, always a product consisting of
comminuted tobacco, either ground or fine-cut.


 Page 453

Fig. 14.1
Smokeless tobacco products, organized
according to mechanical processing.
One attempt to classify the smokeless tobacco category based upon
the manner in which the tobacco is disintegrated is shown in Fig. 14.1.
Products And Prevalence
People in all parts of the world have used tobacco in different forms
ever since Columbus and his crew introduced this mysterious, 'health-
bringing' plant to the old world in the late fifteenth century. The
American Indians at that time used tobacco in all ways that we know
of today, inhaling smoke from burned tobacco, sniffing small particles
into the nose or introducing a fair amount of tobacco into the mouth.
The early product types were, of course, made by hand and the
tobacco was processed to provide the shape required for the intended
end use. Development of manufactured products came later with the
industrial revolution during the nineteenth century. Even today, large
quantities of smokeless tobacco are produced with simple manual
methods in rural areas of the world.
Smokeless tobacco has its strongholds in all large continents. The
description below is not in any way complete because of the simple
fact that statistics do not exist for most countries in the Third World.
a
Europe
In northern Europe, especially in Sweden, the per capita consumption
of oral snuff is among the highest in the world. Approximately 18% of
adult Swedish males use oral snuff. Swedish snus is a ground tobacco
product, moisture content 50 to 60%, which is normally taken as a
pinch with the fingers and placed in the vestibular area of the upper
jaw. An alternative packaging form is the portion-packed snus, i.e. a
portion (0.5 or 1 g) packed in a porous paper-like material. Both snus
types exist in a variety of flavors.
Products similar to the ones in Sweden are used in Norway, Denmark,
Finland and Germany, although the habit is not as widespread as in
Sweden. The Scandinavian type of snuff is more or less unknown in
other parts of Europe. An oral snuff named Makla is sold in France.
This is used, however, by North Africans, primarily immigrated
Algerians.
There is a small but steady use of nasal snuff in many European
countries. The largest usage is found in Germany (especially in
Bavaria) and in the British Isles. The nasal snuff is generally a finely
ground tobacco, dry or semi-moist, offered in a lot of aroma varieties.
A very special type is the Bavarian Schmalzler, which is moistened
with paraffin oil.
Various types of chewing tobacco (twist, plug or pressed tobacco
portions) are found in many European countries. The use of these
types is, however, steadily declining.
b
North America
On a volume basis, the USA has the highest use of smokeless tobacco
in the world. Several product types are found on the market, the
largest being loose leaf and oral moist snuff. Loose leaf, which is
often referred to as just chewing tobacco in the consumer's
vocabulary, is a special product made for the USA. It is made from
flakes of threshed tobacco leaves which are generally heavily
flavored. Other types of smokeless tobacco found in the USA are dry
snuff (also called Scotch type snuff, used orally or nasally), plug
chewing tobacco (compressed blocks of tobacco) and twist (spun
strips of tobacco).
The most common way of using moist snuff in the USA is to place the
snuff portion in the lower part of the mouth between the lower lip and
the teeth.
A limited quantity of American types of smokeless tobacco is sold in
Canada.
c
Africa
Many African countries have a very high use of smokeless tobacco.
The products are often handmade and exist in various forms. The
usage is not well explored and consumption statistics are nonexistent.
Algeria has an industrial production of both nasal and oral snuff made
from tobacco grown in North Africa. Similar products are found in
Tunisia, Libya and Egypt. Toombak, which is a moist snuff product
for oral use, has a very high use rate in Sudan; it has been

 Page 454
estimated that as many as 23% of the Sudanese population use this
type of product (Idris, et al., 1991). Nigeria has a long traditional use
of both oral and nasal snuff products, a large part of which is
produced from imported black fat, a specially treated air-cured
tobacco imported from the USA. Various types of snuff and chewing
tobacco are used in South Africa.
d
Asia
India and Pakistan are the largest consumers of smokeless tobacco
products in Asia. A broad selection of industrially manufactured
products is found, designed for either nasal or oral use. A special
chewing tobacco type is the zafrani patti, which is a dry particulated
tobacco, highly scented and containing fine flakes of metallic silver.
In many parts of Asia the combined chewing of tobacco and betel leaf
or areca nut is common.
Various forms of smokeless tobacco are found in other Asian
countries, e.g. Afghanistan, Saudi Arabia and the former Soviet
Union. For example, nass is a smokeless tobacco product widely used
in some of the former Soviet Republics. It is described as a mixture of
tobacco, lime, ash and cotton seed oil (Zaridze, et al., 1991).
Tobaccos Used
Moist snuff is primarily made from fire-cured and air/sun-cured dark
tobaccos. Large producers of these types of tobacco are the USA
(about 26 000 tons/year in 1994), Poland (about 12 000 tons/year),
Italy (about 25000 tons/year), Malawi (about 8000 tons/year),
Tanzania (about 4000 tons/year) and South Africa (about 4000
tons/year). India has the largest production of sun-and air-cured dark
tobaccos (about 350 000 tons/year) in the world. This tobacco is used
in the domestic manufacture of snuff, chewing tobacco and smoking
products. Some is exported to other countries for use in smokeless
tobacco products.
Tobacco grown in the USA is the predominant raw material for moist
snuff in the Western hemisphere. Domestic tobaccos and tobaccos of
other provenances are, however, also of importance. Thus, German,
Brazilian and Hungarian tobaccos have found use in certain products.
The North African manufacturers use only domestic and Pakistani
tobaccos, essentially of Nicotiana rustica type. Tunisia, for instance,
has an annual production of sun-cured N. rustica of about 800 tons,
some of which is exported.
Pennsylvania and Wisconsin tobaccos are the raw materials of choice
in American loose leaf products, the production in 1994 of these
tobaccos being some 4000 and 3000 tons, respectively.
Low grade tobacco, tobacco dust and stems may be used in a low
percentage in snuff blends. Flue-cured tobaccos are generally not
suited for use in oral smokeless tobacco products because of their
bitter taste. If traditionally flue-cured varieties are air-cured, the bitter
taste is avoided and the resultant tobacco may find use as ingredients
in various products.
Since the bulk of the dark tobacco produced, at least in the USA, is
nowadays used in smokeless tobacco products and not in smoking
products, it is worthwhile to include a discussion on the characteristics
of such tobacco.
Dark Tobacco Growth, Curing and Classification
Dark tobaccos owe the dark green color of their leaves to the high
chlorophyll content, the level being twice as high as that of burley
tobacco measured on an area basis (Griffith, et al., 1944). These
tobaccos are generally grown on heavy soils with wide spacings
between the plants, a heavy nitrogen fertilization regime being
applied. The plants are topped early, 30 to 40 days before maturity, in
order to insure growth of large leaves with a gummy texture and a
heavy body. Harvesting takes place most commonly with stalk cutting
and the tobacco is either fire-cured or air-cured (Tso, 1990).
Additional details on dark fire/air-cured tobacco production are
provided in Chapter 5D of this monograph.
Smoke from hardwood fires, e.g. hickory smoke, is used in the fire-
curing process. The tobacco, which is hung in closed barns, undergoes
a relatively slow yellowing phase. If adverse weather conditions
prevail, small fires may be started to aid this phase. When yellowing
is complete and brown spots begin to appear on the leaves, the
temperature is increased by using heat from small fires. The humidity
is slowly reduced. When the leaves have developed a brown color,
they are exposed to large amounts of smoke for 2 to 3 weeks. The
smoke is produced by adding wet sawdust to the fires, which are kept
low in order not to provide undue heat. The resultant cured leaves are
dark and coated with wood smoke condensate (Tso, 1990).
In the air-curing process, the tobacco is not exposed to wood smoke.
The stalk-cut tobacco plants are hung in a barn and slowly cured over
a period of several weeks. The cured leaf is heavy and thick and has a
dark brown color (Tso, 1990).

 Page 455
There are two basic types of dark tobacco grown in the USA: broad
leaf types and onesucker types. The broad leaf types, exemplified by
KY 171 and Madole, may be either air-cured or firecured, while most
onesucker types, e.g. KY 160, are air-cured. Dark firecured tobacco is
designated as class 2 and subdivided into types 21 to 23, whereas dark
air/sun-cured tobacco belongs to class 3B and types 35 to 37 (Tso,
1990) (see Table 14.1).
Table 14.1 USA production (1994) in tons of
firecured and air-cured and air-cured
tobaccos.
Type 21 Virginia firecured 1 000
Types 22 to Kentucky Tennessee 20
23 firecured 000
Type 35 Onesucker 3 300
Type 36 Green River 1 500
Type 37 Virginia sun-cured 50

Dark Tobacco Chemical Composition

In contrast to flue-cured, burley and Oriental tobaccos, which have
been subjected to thorough chemical studies, published information
on the chemical composition of dark tobacco is sparse. The
presentation below on some chemical characteristics of dark tobacco
is based on data collected from the literature and on results of
unpublished studies.
Although the levels are dependent upon factors such as weather
conditions, fertilization rates, fertility and topping heights, dark
tobaccos are generally fairly rich in nicotine. Average values may
range between 3.5 and 4.0% for American varieties. Chamberlain, et
al. (1988) found in a 1-year study that the concentration of nicotine
was higher in air-cured KY 171 (4.5%) than in firecured KY 171
(3.4%) and air-cured KY 160 (2.9%). They also observed that the
levels of certain other constituents such as malic acid (5.0 to 8.9%)
and the polyisoprenoid solanesol (1.8 to 2.8%) were considerably
higher in these dark tobaccos than in a reference flue-cured variety
(NC 2326; 1.9 and 1.2%, respectively). As expected, the concentration
of the polyphenol chlorogenic acid is low in the dark air-and firecured
tobaccos (0.05 to 0.1%).
Dark tobacco synthesizes starch in considerable amounts. The level is
high at harvest (some 30%), but decreases rapidly during curing due
to hydrolysis (unpublished data). Virtually no starch and very low
levels of sugar are found at the end of cure.
The content of nitrate, nitrite and tobacco-specific nitrosamines
(TSNA) in tobacco and particularly in dark varieties has received
special attention. The nitrate levels show considerable variation
between crops. In general, lamina from the bottom position contain
higher concentrations of nitrate than lamina from upper stalk positions
(Davis, et al., 1981; Burton, et al., 1994). As is the case with other
types of tobacco, the midvein is high in nitrate; typically 5 to 10 times
higher nitrate levels are found in the midvein compared to the lamina
(Burton, et al. 1994). Burton, et al. (1992) studied the distribution of
certain tobacco constituents within the tobacco leaf (KY 171) and
found that the nitrate levels are highest next to the midvein and lowest
at the outer edges of the lamina. The tip of the leaf contains the very
lowest concentration.
Nitrite, which may accumulate during curing as a result of bacteria-
mediated reduction of nitrate (Burton, et al., 1989), is commonly
present in trace amounts (µg/g) in sound, cured tobacco. The base of
the leaf (lamina and midvein) was found to contain the highest
concentration of nitrite, whereas the lowest amounts were present at
the tip of the leaf. Lamina close to the mid-rib was lower in nitrite
than the surrounding lamina segments (Burton, et al., 1992). It is
noteworthy that TSNA has a distribution similar to that of nitrite. By
contrast, alkaloids are most plentiful on the periphery of the leaf, with
the midvein having the lowest concentrations.
The TSNA levels in commercially available dark firecured and dark
air-cured tobacco show extensive variations. The levels are most
certainly dependent upon environmental factors, the curing method
applied and the tobacco genotype used. In a 1-year experiment under
controlled conditions, Chamberlain, et al., 1988 observed that the
concentration of nitrosonornicotine was higher in a firecured sample
of KY 171 than in an air-cured sample of this genotype. Burton, et al.
(1994) found that an air-cured flue variety (Speight G 28) produced
consistently lower quantities of TSNA than a burley (KY 14) and a
dark variety (KY 171) when grown and air-cured under identical
conditions. Our studies on air-cured, flue-cured and dark varieties
support these results.
The volatile fraction obtained from an American dark firecured
tobacco is an extremely complex mixture consisting of 'genuine'
tobacco constituents as well as components derived from the
hardwood smoke deposited onto the tobacco leaf during the firecuring
process (Nordfors & Wahlberg 1989, unpublished data). Thus, of the
300 neutral compounds identified by gas chromatographymass
spectrometry, more than 150 are derived from the tobacco itself and
some 140 are derived from the hardwood smoke. The former

 Page 456
group includes regular mono-, sesqui-and diterpenoids, compounds
formed by biodegradation of carotenoids, acyclic isoprenoids and
cembranoids as well as fatty acid-and phenylalanine-derived
compounds. Compounds representing each of these biogenetic groups
are listed in Table 14.2. Most, if not all, of these compounds have
previously been reported as constituents of other tobaccos and several
are well known aroma constituents (Demole & Berthet, 1972a; Lloyd,
et al., 1976; Fujimori & Kaneko, 1979; Wahlberg & Enzell, 1987). It
is noteworthy that diterpenoids of the labdane class are not produced
by the American dark tobacco. This may not, however, be true for
dark tobaccos of other provinces.
The neutral constituents originating from the hardwood smoke and
identified in this study comprise alkyl-substituted naphthalenes,
substituted di-and tetrahydronaphthalenes, methyl-and
dimethylsubstituted biphenyls, alkyl-substituted 2,3dihydroinden-1-
ones, methoxybenzenes and benzofurans. Several pyrroles and furans
were also encountered. Compounds of the latter two groups are
commonly classified as Maillard products. Some of them are likely to
originate from the wood smoke, while others may be tobacco
constituents.
Some 80 compounds were identified in the fraction containing weak
acids, alkylphenols, guaiacols and syringols being the major
components. Cyclotenes and
Table 14.2 Volatile components (examples)
identified in an extract of an American
dark fire-cured tobacco and listed
according to plausible biogenetic origin.
Monoterpenoids:
Linalool
Tetrahydrolinalool
2,6-Dimethyl-2,7-octadiene-l,6-diol (2
isomers)
Carvenone
Piperitone

Sesquiterpenoids:
Nerolidol
Solavetivone
Occidol

Diterpenoids:
Neophytadiene
Geranylgeranadiene
Phytol
Dihydrophytol
Cembrene

Degraded carotenoids:
1,1,3-Trimethyl-3-cyclohexene-2,4-dione
1,1,3-Trimethyl-2,4-cyclohexanedione
1,1,3-Trimethyl-2-cyclohexene-4,5-diol
2-Formyl-1,1,3-trimethyl-2-cyclohexen-4-
one
Dihydroactinidoilide
3-Oxodihydroactinidiolide
ß-lonone
ß-lonol
4,6,8-Megastigmatrien-3-one (4 isomers)
ß-Damascone
Damascenone
5,6-Epoxy-ß-ionone
3-Hydroxy-ß-ionone
4-Hydroxy-ß-ionone
3-Hydroxy-5,6-epoxy-ß-ionone
4,5-Epoxy-a-ionone
6-Hydroxy-a-ionone
3-Oxo-a-ionol
(table continued on next page)

 Page 457
Contd
Table 14.2
3-Oxo-7,8-dihydro-a-ionol
3-Oxo-6-hydroxy-a-ionol
3-Hydroxy-ß-damascone
1,3,7,7-Tetramethyl-2-
oxabicyclo[4.4.0]dec-5-en-9-one (2
isomers)
1,3,7,7-Tetramethyl-2-
oxabicyclo[4.4.0]decan-9-one (2 isomers)

Degraded acyclic isoprenoids:

6-Methyl-3,5-heptadien-2-one
Geranylacetone
Pseudo-ionone (2-isomers)
2,6-Dimethyl-2-hydroxy-3,6-undecadien-
10-one
Farnesylacetone
Hexahydrofarnesylacetone

Degraded cembranoids:
Norsolanadione
5-lsopropyl-2,8-nonanedione
5-lsopropyl-3-nonene-2,8-diol
5-lsopropyl-2,8-nonanediol
2-Acetonyl-3-isopropyl-6-
methyltetrahydropyran (3 isomers)
Solanone
Solanol
1,4-Epoxy-7-isopropyl-4-methyl-5-
undecen-10-one
Prenylsolanone

Fatty acid-derived compounds:

2-Tridecanone
2-Pentadecanone
3Z-Hexen-1-ol
3-Hexanol
Octanol
Nonanol
Decanol
Undecanol
Dodecanol
Tridecanol
Tetradecanol
Pentadecanol
Hexadecanol
1 -Octen-3-ol
3-Nonen-1-ol
1,9-Nonanediol
4-Butanolide
4-Octanolide
4-Nonanolide
5-Octanolide
Bovolide
Dihydrobovolide
Spiroxabovalide
Methyl nonanoate
Methyl hexadecanoate

Phenylalanine and lignin-derived

compounds:
Benzaldehyde
Benzyl alcohol
2-Phenylethanol

 Page 458
maltols are minor components. These compounds, which are largely
derived from the hardwood smoke, give important contributions to the
smoky, spicy, woody and sweet flavor of the fire-cured tobacco.
Syringols, which are noted for their desirable aroma properties, are
abundant in smoke from hardwood, whereas smoke from needle-
leaved trees contains a larger proportion of guaiacols (Gilbert &
Knowles, 1975). The type of wood used in the curing process will
therefore have a substantial impact on the aroma of fire-cured
tobacco.
The acidic fraction was found to contain normal (C2 to C16) and
branched saturated and unsaturated fatty acids of the types commonly
present in tobacco (Demole & Berthet, 1972b; Lloyd, et al., 1976;
Chuman & Noguchi, 1977; Wahlberg, et al., 1977a). This fraction is
also fairly rich in methyl-and dimethyl-substituted benzoic acids and
in furoic acids. These compounds are most likely derived from the
hardwood smoke.
In addition to nicotine, the basic fraction examined contains nicotine
metabolites such as nicotine N-oxide, b-nicotyrine, 2,3'-bipyridyl and
cotinine. Other alkaloids found were N-methylanabasine, N-
methylanatabine and myosmine. All these compounds are genuine
tobacco constituents (Schmeltz & Hoffmann, 1977; Wahlberg, et al.,
1977b), no smoke-derived compounds being detected among the
bases in this fraction.
Several of the compounds mentioned above have been reported by
Davis, et al. (1981), who examined the essential oils obtained from a
dark tobacco that was either fire-cured or aircured. As expected,
certain phenols and aromatic hydrocarbons were only detected in the
fire-cured tobacco.
Manufacturing Methods
Several vastly different manufacturing methods are or have most
certainly been applied to produce smokeless tobacco products. The
discussion below comprises only manufacture of a few of the products
that exist today. The descriptions cannot be detailed, since
manufacturers do not generally reveal their manufacturing methods.
Nor can the production methods for products from the Third World be
described due to a general lack of documentation.
Manufacture of smokeless tobacco includes some general production
steps. These may, however, vary in the order of performance:
selection of cured leaf tobaccos suitable for the end product;
post curing processing of leaf tobacco;
leaf tobacco disintegration, i.e. stripping, threshing, cutting or
grinding;
blending of tobaccos;
processing of blended tobacco;
finishing of product, addition of functional additives like flavors and
stabilizers.
a
Loose Leaf Chewing Tobacco, USA
As mentioned above, Pennsylvania and Wisconsin aircured tobacco
leaves are generally used as raw material for loose leaf products. The
cured tobacco is subjected to 'sweating' at a slightly elevated
temperature for a certain period of time. The tobacco leaves are then
threshed into flakes and the stems (mid-ribs) are removed. The
tobacco fragments thus obtained are usually treated with a sweet
casing solution, dried and packed in the consumer package.
b
Plug Chewing Tobacco, USA
Plug tobacco is usually a compressed form of loose leaf, where a
certain amount of finished loose leaf tobacco is charged into a mold
and pressed to acquire a flat bar shape. Usually, the tobacco bar is
wrapped with either natural tobacco or reconstituted sheet tobacco.
Two forms of plug tobacco exist, dry and moist plug.
c
Moist Snuff, USA
After loose leaf, moist snuff is the largest smokeless tobacco product
on the USA market. Fire-cured and aircured dark tobaccos are mainly
used. The cured tobacco is allowed to age for at least a year before
being taken into production. The USA type of moist snuff is
commonly made from fine-cut tobacco. Different cutting sizes
produce different types of products, e.g. fine cut, coarse cut or long
cut. After cutting, the tobacco is mixed with water and various
ingredients and allowed to ferment in closed vessels for a period of
several weeks. The fermentation is usually controlled by monitoring
pH and temperature.
In the finishing stage after fermentation, further additives are admixed
to the snuff in order to make it stable and to impart a desired flavor.
c
Moist Snuff, Sweden
Since no tobacco is grown in Sweden, imported aircured and fire-
cured tobaccos are used. Swedish snus is usually made from ground
tobacco as opposed to the

 Page 459
USA type of snuff which is generally cut. A heat treatment process is
used. This process combines the effects of sterilizing the snuff blend
and creating the typical snuff aroma and taste. The development of the
snuff aroma has been documented by chemical studies. These have
shown that the concentration of a considerable number of desirable
aroma compounds arising by degradation of carotenoids and
cembranoids increases significantly during the snus process (Nordfors
& Wahlberg 1990, unpublished data). Since many of these compounds
are also generated during aging of tobacco, it may be inferred that the
snus process can replace the aging process. Syringols, cyclotenes and
furoic acid are largely lost during the process. In the final stage of
production, flavors are added. Snus products, additives and hygienic
requirements of production are regulated under the Swedish food
laws.
e
Nasal Snuff, Germany, England
The traditional Bavarian Schmalzler snuff is made primarily from
Brazilian tobacco which is mixed with a casing solution and
fermented for several months at elevated temperature. After being
dried, the tobacco is ground to a fine powder and remoistened with
paraffin oil.
Other types of snuff are produced using a quick method, in which a
blend of tobacco is pulverized, mixed with a casing of flavors and
allowed to ferment for 6 to 8 weeks. The snuff is then sieved and
some further casing is added. This is the predominant snuff making
method today in both Germany and England. Smokeless tobaccos are
regulated by the German Tobacco Ordinance in Germany.
f
Moist Snuff, Sudan
Sudanese snuff (saffa or toombak) is made from suncured tobacco
leaves, mainly locally grown Nicotiana rustica. The leaves are mixed
with an aqueous solution of sodium carbonate and kept for
approximately 24 hours in a closed container before they are
converted to snuff. The snuff production is mainly done by hand
(Idris, et al., 1991).
Chemical Composition of Smokeless Tobacco Products
Published information on the chemical composition of smokeless
tobacco products is largely focused upon the content of controversial
compounds such as TSNA and polycyclic aromatic hydrocarbons
(PAH). It should be noted, however, that such data are available only
for very few of the numerous types of smokeless tobacco products
existing around the world today. It should also be kept in mind that the
chemical composition of a smokeless product is dependent upon the
ingoing tobaccos and additives, manufacturing methods and storage
conditions. Therefore, wide variations with respect to the
concentration of many chemical constituents are expected in various
product types.
Hoffmann, et al. (1991, 1994) have compared the TSNA levels in a
few selected smokeless tobacco products. They found that the levels
are lower in USA loose leaf products than in USA and Swedish moist
snuff. Also, they noted that the concentration of TSNA in the major
commercial brands in the USA and in Sweden, 20.8 to 22.5 µg/g and
9.3 to 11.4 µg/g, respectively, have declined significantly during the
past 10 years and they relate this decrease to improvements in
manufacturing methods. The markedly low amounts of TSNA, 0.19 to
0.95 µg/g, detected in Soviet nass have also been attributed to the
manufacturing method which involves a short aging process (Zaridze,
et al., 1991). Sudanese toombak, which has exceptionally high levels
of TSNA, represents another extreme. Both the manufacturing process
applied and the tobacco used have been considered as contributing to
the TSNA accumulation (Idris, et al., 1991).
Trace amounts of PAH may be encountered in smokeless tobacco
products. Thus, five USA snuff brands examined in 1985 displayed
levels of benz(a)pyrene ranging from 0.1 to 63 ng/g on a dry weight
basis (Hoffmann, et al., 1987). The levels of these and other wood
smoke derived compounds are most certainly associated with the
proportion and origin of the ingoing tirecured tobacco, hence
explaining the dramatic variation among smokeless tobacco products.
Of the polyphenols, chlorogenic acid was not detected or was present
in a small amount in five USA moist snuffs studied in 19856
(Hoffmann, et al., 1987). Rutin and kaempferol are present in amounts
ranging from 0.2 to 2.6%. Higher concentrations of these polyphenols
are found in the three dry snuff products also examined. It is
noteworthy that these compounds, which represent a major portion of
the water extractables, are antioxidants and antimutagens. a-
Tocopherol (vitamin E), a fat-soluble antioxidant, is another tobacco
phenol that is present in a concentration of about 15 µg/g in Swedish
snus.
The concentration of nicotine varies among smokeless products. Since
N. rustica is rich in nicotine, it is not surprising to find products
containing 4 to 9% of

 Page 460
nicotine (dry weight) among Sudanese snuff samples (Idris, et al.,
1991). By contrast, nasal snuff products low in nicotine are found for
instance in Europe, and low nicotine moist snuff brands have been
introduced in both the USA and Swedish markets in recent years.
The additives frequently used in smokeless products may influence
the chemical composition to a large extent. For example, sodium
chloride is often added to moist snuff and to some dry snuffs for nasal
consumption. Finished products may contain up to 7 to 10% by
weight of sodium chloride, which gives a desired salty taste to the
snuff. Ammonium chloride is sometimes used instead of, or in
combination with, sodium chloride to produce the special salmiac
taste.
Sugar and sweeteners such as saccharin and sorbitol are frequently
found in smokeless tobacco products. Chamberlain, et al. (1988), who
have screened a series of USA commercial smokeless products,
reported that as much as 20% of sucrose is present in certain chewing
tobaccos (loose leaf products).
Benzyl benzoate, a flavor fixative, is sometimes used, and levels in
commercial snuff brands have been estimated to range between 30
and 110 µg/g of tobacco (LaVoie, et al., 1989). Glycerol, 1,2-
propylene glycol and other conditioners are found in certain products.
Flavors are commonly added to smokeless tobacco products. While
wintergreen oil and other essential oils are traditional additives, new
types of alkali-stable flavoring materials have found use in snuff
products.
References
Burton, H.R., Childs, G.H., Andersen, R.A. & Fleming, P.D. (1989)
Changes in chemical composition of burley tobacco during
senescence and curing. 3. Tobacco-specific nitrosamines. J. Agric.
Food Chem., 37, 42630.
Burton, H.R., Dye, N.K. & Bush, L.P. (1992) Distribution of tobacco
constituents in tobacco leaf tissue. 1. Tobacco-specific nitrosamines,
nitrate, nitrite, and alkaloids. J. Agric. Food Chem., 40, 105055.
Burton, H.R., Dye, N.K. Bush, L.P., (1994) Relationship between
tobacco-specific nitrosamines and nitrite from different air-cured
tobacco varieties. J. Agric. Food Chem., 42, 200711.
Chamberlain, W.J., Schlotzhauer, W.S. & Chortyk, O.T. (1988)
Chemical composition of nonsmoking tobacco products. J. Agric.
Food Chem., 36, 4850.
Chuman, T. & Noguchi, M. (1977) Acidic aroma constituents of
Turkish tobacco. Agric. Biol. Chem., 41, 102130.
Davis, D.L., Song, B.H. & Weeks, W.W. (1981) Chemical
composition of dark air-cured and fire-cured tobacco. Paper presented
at 35th Tob. Chem. Res. Conf., Winston-Salem, NC (abstract 30).
Demole, E. & Berthet, D. (1972a) A chemical study of burley tobacco
flavor (Nicotiana tabacum L.). I. Volatile to mediumvolatile
constituents (b.p. <84°/0.001 Torr). Helv. Chim. Acta, 55, 186682.
Demole, E. & Berthet, D. (1972b) A chemical study of burley tobacco
flavor (Nicotiana tabacum L.) II. Mediumvolatile, free acidic
constituents (b.p. 84-114°/0.001 Torr). Heir. Chim. Acta, 55,
18981901.
Fujimori, T. & Kaneko, H. (1979) Studies on tobacco aroma. Nippon
Nogeikagaku Kaishi, 53, R95R121.
Gilbert, J. & Knowles, M.E. (1975) The chemistry of smoked foods: a
review. J. Food Technol., 10, 24561.
Griffith, R.B., Vaileau, W.D. & Jeffrey, R.N. (1944) Chlorophyll and
carotene content of eighteen tobacco varieties. Plant Physiol., 19,
68993.
Hoffmann, D., Adams, J.D., Lisk, D., et al. (1987) Toxic and
carcinogenic agents in dry and moist snuff. JNCI., 79, 12816.
Hoffmann, D., Brunnemann, K.D., Prokopczyk, B. & Djordjevic,
M.V. (1994) Tobacco-specific N-nitrosamines and Areca-derived N-
nitrosamines: chemistry, biochemistry, carcinogenicity, and relevance
to humans. J. Toxic. Environ. Health, 41, 152.
Hoffmann, D., Djordjevic, M.V. & Brunnemann, K.D. (1991) On the
control of toxic substances in smokeless tobacco. J. Smok.Rel. Dis., 2,
16572.
Idris, A.M., Nair, J., Ohshima, H., et al. (1991) Unusually high levels
of carcinogenic tobacco-specific nitrosamines in Sudan snuff
(toombak). Carcinogenesis, 12, 111518.
LaVoie, E.J., Tucciarone, P., Kagan, M., et al. (1989) Analyses of
steam distillates and aqueous extracts of smokeless tobacco. J. Agric.
Food Chern., 37, 1547.
Lloyd, R.A., Miller, C.W., Roberts, D.L., et al. (1976) Fluecured
tobacco flavor. I. essence and essential oil components. Tob. Sci., 20,
4048.
Schmeltz, I. & Hoffmann, D. (1977) Nitrogen-containing compounds
in tobacco and tobacco smoke. Chem. Rev., 77, 295311.
Tso, T.C. (1990) Production, Physiology, and Biochemistry of
Tobacco Plant. Ideals, Inc, Beltsville, Maryland.
Wahlberg, I. & Enzell, C.R. (1987) Tobacco isoprenoids. Nat. Prod.
Rep., 4, 237276.
Wahlberg, I., Karlsson, K., Austin, D.J., et al. (1977a) Effects of flue-
curing and aging on the volatile neutral and acidic constituents of
Virginia tobacco. Phytochem. 16, 121731.
Wahlberg, I., Karlsson, K., Austin, D.J., et al. (1977b) Effects of flue-
curing and aging on the volatile basic constituents of Virginia tobacco.
Phytochem., 16, 12335.
Zaridze, D.G., Safaev, R.D., Belitsky, G.A., et al. (1991)
Carcinogenic substances in Soviet tobacco products. In: Relevance to
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Publication 105, Lyons.

 Page 461

Index
A
Acrosternum hilare, 237
aging, 19, 273
Agrobacterium tumefaciens, 5, 53
Agrotis ipsilon, 231
alkaloid accumulation, 286
leaf distribution, 287
alkaloid biosynthesis, 285
genetic control, 286
nicotine synthesis, 287, 288, 289
nitrogen fertility, 285
secondary alkaloid synthesis, 289, 290
Alternaria alternata, 38, 185
amadori compounds, 274, 307
American blended cigarettes, 105
amino acids, 273, 307
ammonia, 270, 273
ammonia/smoke, 420
anabasine, 285
anatabine, 285
Aphelenchoides spp., 220
aromatic acids, 309
ash, 282
B
Bacillus thuringiensis, 5, 56
bacterial diseases, 187
bacterial wilt (Granville wilt), 195
description, epidemiology, management, 196
hollow stalk, 188
description, epidemiology, management, 188, 189
wildfire and angular leaf spot, 187
description, epidemiology, management, 187, 188
bacterial wilt, 38, 195
Bemisia tabaci, 239
biotechnology, 49
AFLP, 52
Agrobacterium mediated transformation, 53
genetic linkage map, 50
herbicide resistance, 54
insect resistance, 56
molecular marker, 50
 nematode resistance, 57
RAPD, 51
RFLP, 50
stress tolerance, 58
transformation, 52
virus resistance, 58
black root rot, 37, 173, 191
black shank, 37, 173, 189
blending, 21, 346
blue mold, 37, 157, 174, 183
boron, 85, 114
breeding and genetics, 32
advanced line testing, 45
chemical constituents, 36
chlorophyll level, 35
disease resistance, 36
disease screening, 44
evaluation of genotypes, 44
field testing, 45
germplasm, 33
inheritance of characters, 33
insect resistance, 39
 methods, 41
backcross method, 42
doubled haploid, 43
genetic engineering, 44
interspecific hybridization, 43
morphological traits, 33
pedigree method, 42
pure-line, mass selection, 42
recurrent selection, 43
yield and quality, 40
burley tobacco, 143, 266, 276, 347, 376
burn modifiers, 359
burn rate, 317
factors, 317
measurements, 317
C
calcium, 84, 114
deficiency, 84
Cambridge filter pad, 399
candela wrapper, 448
carbohydrates, 266, 270, 306
carbon dioxide, 46, 417
carbon monoxide, 417
carotenoids, 275, 276, 277, 309, 310
casings and flavorings, 350, 351
cellulose, 268, 269, 306

 Page 462
cellulose acetate, 363, 364
cembranoids, 278, 294, 295, 296
charcoal filters, 365
chemical composition of mainstream smoke, 416
chemical ETS marker, 429
chloride, 86, 118, 171
tobacco quality, 86
chlorophyll, 275
cigar binder, 441
cigar filler, 441
cigar wrapper, 441
cigar/definition, 440
cigarette beetles, 241, 243
action threshold, 244
freezing, 245
fumigation, 247, 341, 342, 345
heat, 245
housekeeping/sanitation, 244, 344
insect growth regulators, 246, 247
insecticides, 246
pheromone trap monitoring, 243, 345
cigarette burn rates, 317, 318
cigarette circumference/QA, 393
cigarette design and materials, 353
brand proliferation, 355challenges, 381
cigarette environmental degradability, 382
cigarette filters, 363, 373
cigarette paper, 359, 360, 361, 362, 363
cigarette paper composition, 359
cigarette smoke and smoke formation, 355, 356
cigarette testing, 356, 357, 358, 359
environmental tobacco smoke, 382, 383
expanded tobacco, 377, 379, 380
filters/cigarette performance, 365, 366, 367, 368, 369
reconstituted sheet, 377, 378, 379
reduced smoke yields, 354, 355
tobacco rod, physical characteristics, 380, 381
tobacco types and blends, 375, 376, 377, 378
cigarette filter plasticizer/QA, 396
cigarette firmness/QA, 395
cigarette length/QA, 396
cigarette moisture/QA, 395
cigarette pack seal testers, 397
cigarette paper porosity/QA, 395
cigarette pressure drop/(RTD)/QA, 394
cigarette ventilation/QA, 394
cigarette weight/QA, 393
cigarette/air flow measuring instruments, 396
cigars and cigarillos, 440
agronomics, 442
binder, 441
farming practices, 442
fermentation, 449
filler, 441
production practices, 443
sorting, 451
varieties, 442
wrapper, 441
classification of tobacco, 1
botanical characterization, 2
chemical composition, 3
climatic factors, 10, 92
carbon dioxide, 96
humidity, 92
light, 95
 temperature, 93
codex alimentarius commission, 256
combustion, 400
contact, local systemic suckercides, 122
contact suckercides, 121
contact systemics, 149, 177
crop rotation, 144, 170
curing, 17, 129, 151, 159, 178, 446
curing-energy sources, 130
cutin, 298
cyst nematodes, 40
D
dark fire-cured tobacco, 164, 454
chemical composition, 455
classification, 454
disease control, 173
harvesting and curing, 178, 454
insect control, 175
market preparations, 180
site selection and fertilization, 170
topping and sucker control, 176
transplant production, 166
 transplanting and plant spacing, 172
variety selection, 165
weed control, 171
dark tobacco curing stages, 179, 454
dinitroanalines, 122
direct seeding, 167
diterpenes, 294, 295, 296, 308
Ditylenchus dipsaci, 220
E
environmental influences/tobacco production, 7
environmental tobacco smoke constituents, 406, 428, 430
Ephestia elutella, 242
Epitrix spp., 234

 Page 463
equilibrium moisture, 314
adsorption isotherm equation, 315
content, 315
flavoring effects, 316
measurement, 315
pore size/specific surface area, 316
Erwinia carotovora, 188, 448
Erysiphe cichoracearum, 186
Euschistus servus, 237
expanded tobacco, 20, 318, 377, 378, 379, 380
F
fatty acids, 309
fatty alcohols, 121, 149, 177
fermentation, 19, 161, 162, 338, 449
fertilizers, 13, 120, 145, 444
field practices, 76
nutrition, 79
plant populations, 77
seedling establishment, 76
topping and suckering, 86
filler grades, 143, 346
filling values, 313
factors, 314
measuring methods, 313
filter materials and specifications, 364
cellulose acetate filters, 364
cigarette performance, 365
filter pressure drop, 364
filter ventilation, 369, 370, 371
fines, 336
firmness, 358
flavor grades, 346
float system, 167, 169, 443
flower development, 66
flue-cured, 104, 266, 274, 346, 376
curing and marketing, 129
fertilizer application, 120
future, 136
harvesting, 126
marketing, 136
seedling production, 107
soil properties, 113
topping and sucker control, 120
 weed control, 109
flue-curing process, 131
chlorophyll degradation, 131
protein hydrolysis, 131
starch hydrolysis, 131
foliar insect pests, 233
aphids, 233
budworms, 234
flea beetles, 234
grasshoppers, 235
hornworms, 235
laceworms, 236
loopers, 236
stink bugs, 237
thrips, 237
tobacco leaf-miner (spitworm), 238
tobacco slug, 238
tobacco stem borer, 239
whiteflies, 239
Frankliniella spp., 237
free radicals/smoke, 427
Frenching, 7
fumigation, 247, 341, 342, 345
fungal diseases, 183
black root rot, 191
description, epidemiology, management, 191, 192
blank shank, 189
description, epidemiology, management, 189, 190
blue mold, 183
description, epidemiology, management, 184, 185
brown spot, 185
description, epidemiology, management, 185, 186
collar rot, 195
description, epidemiology, management, 195
fusarium wilt, 194
description, epidemiology, management, 194, 195
powdery mildew, 186
description, epidemiology, management, 186, 187
sore shin and damping off, 192
description, epidemiology, management, 192, 193
target spot, 193
description, epidemiology, management, 193, 194
Fusarium oxysporum, 194
fusarium wilt, 37, 194
G
gas phase/smoke aerosol, 412
genetic mapping, 49
genome analysis, 49
Globodera tabacum, 219
Gonocephalum, 231
good agricultural practices, 250
H
harvesting, 126, 150, 158, 178, 445
Heliothis, 234
herbicide resistance, 54
hogshead, 338
homogenized leaf curing, 18
hydrazine, 420
I
ignition propensity, 382
inorganics, 282

 Page 464
insect control, 175
insect pests, 228
chemical insecticides, 229
life cycles, 228
insect resistance, 56
integrated pest management, 230, 250, 262
irrigation, 90
isoprenoids, 276
J
Joint FAO/WHO Meeting on Pesticide Residues (JMPR), 256
K
Kabat (methoprene), 246
L
labdanoids, 278, 279, 293, 294, 295, 296
Lasioderma serricone, 241
leaf and smoke components, 305
leaf chemistry, 3, 265
carbohydrates and smoke chemistry, 270
cellulose and hemicelluose, 268, 269
glucosides, 268
nitrogenous constituents, 272
 pectin, 269
phenolics, 281
starches and sugars, 266, 267
sugar esters, 267
tobacco types, 266
leaf chemistry and sensory role, 270, 304
leaf chemistry/organoleptic properties, 304
ammonia, 270
carbohydrates, 270, 306
lipids, 308
nicotine, 271, 310
proteins and amino acids, 307
smoke pH, 271
tobacco alkaloids, 309
leaf classification, 323, 324, 325
leaf physical properties, 313
equilibrium moisture, 314
filling values, 313
leaf pricing and payment, 326, 327
leaf surface chemistry, 292
diterpenes, 294
function, 299, 300, 301
 leaf surface chemicals, 293
nomenclature, 292
sugar esters, 296
surface waxes, 297, 298
volatile compounds, 299
light air-cured tobacco, 143
curing, 151
curing facilities, 151
fertilization practices, 145
harvesting, 150
market preparation, 153
plant populations, 146
site selection and tillage, 144
soil factors, 143
topping and sucker control, 148
transplant production, 146
weed control, 147
lignin, 281, 307
lipids, 308
loose leaf chewing tobacco, 458
M
magnesium, 85, 113
 deficiency, 85
mainstream smoke, 402, 406
maleic hydrazide, 123, 149, 177
maleic hydrazide residues, 125
Manduca quinquemaculata, 235
Manduca sexta, 235
manganese, 114
manganese toxicity, 171
market presentation, 323
market systems, 320
allocation, 321
auction, 320
contract, 321
improvements, 327, 328
regulations, 325
marketing, 129, 153, 180, 348
Maryland tobacco, 143, 266
Meloidogyne arenaria, 217
Meloidogyne hapla, 217
Meloidogyne incognita, 216
Meloidogyne javanica, 216
methoprene, 246
methyl bromide, 166
modifier grades, 346
moist snuff, 458, 459
moisture content, 322
molybdenum, 85, 114
Myzus nicotianae, 233
Myzus persicae, 233
N
nasal snuff, 459
nematode resistance, 57
nematodes, 216
Aphelenchoides spp., 220
control, 221, 222, 223, 224, 225
Ditylenchus dipsaci, 220
Globodera tabacum (tobacco cyst), 219, 220
Meloidogyne spp. (root knot), 216

 Page 465
life cycle, description, distribution, 216, 217, 218,219
Pratylenchus spp., 219
Rotylenchulus spp., 220
Tylenchorhynchus spp., 220
neophytadiene, 277, 278, 308
Nezara viridula, 237
Nicotiana alata, 39
Nicotiana debneyi, 37
Nicotiana excelsior, 37
Nicotiana goodspeedii, 37
nicotiana rustica, 6
Nicotiana sylvestris, 32
nicotiana tabacum, 1, 32
Nicotiana tomentosiformis, 32
Nicotiana velutina, 37
nicotine, 79, 356, 410, 459
nitric oxide/smoke, 420
nitrification, 117
nitrogen, 79, 115
carbohydrate metabolism, 80
growth, yield, quality and chemistry, 80
nitrogen sources, 82, 117
ammonium nitrate, 82
sodium nitrate, 82
urea, 82
nitrosamines, 419, 425, 455, 459
nitrosonornicotine, 18, 425
nornicotine, 285
no-tillage, 144
nutritional disorders, 8
mineral deficiency symptoms, 9
O
oral snuff, 453
organic acids, 280
Oriental tobacco, 154, 266, 348
baling and marketing, 160
cultivation, 157
field preparation, 157
growing regions and types, 154
Basma type, 155
Izmir type, 154
Macedonia Basma type, 155
Samsun type, 156
 harvesting and curing, 158
manipulation and fermentation, 161
seed beds, 157
storage and fermentation, 162
Oulema bilineata, 238
P
paper permeability, 358
particle size distribution-lamina, 333
particulate phase/smoke aerosol, 412
pectins, 269, 307
perforation, 359, 373
Peronospora tabacina Adam, 184
pesticide classification, 253
pesticide registrations, 250, 251,252
pesticide regulations, 250
changing environment, 260
existing maximum residue levels (MRL), 257, 258, 259, 260
international scientific organizations, 255
pesticide residues-analytical methods, 260
pesticide resistance, 261
pesticides, 444
pesticides-industry stewardship, 254
phenolics, 281, 282, 310
phosphine fumigation, 247
phosphorus, 82
deficiency, 83
photoinhibition, 93
photoperiodism, 11
photorespiration, 94
Phthorimaea operculella, 238
physical properties/methods, 313
Phytophthora parasitica, 189
plant beds, 72
clipping, 73
methyl bromide, 73
nutrition, 73
seeding, 73
undercutting, 74
plant populations, 12, 77
chemical composition, 78
leaf characteristics, 78
yield and quality, 78
plant protection agents ADI, 254
plant regeneration systems, 59
plant-water relations, 89
irrigation, 89
soil water deficit, 89
water logging, 90
plasticizers, 364
plug chewing tobacco, 458
polynuclear aromatic hydrocarbons, 423, 459
polyphenols, 281, 422, 459
positional cloning, 60
potassium, 83
deficiency, 83
potyviruses, 39
powdery mildew, 38, 186
Pratylenchus spp., 219
premature flowering, 70, 77

 Page 466
pressure drop, 357
primed seed, 168
primings, 126
processing and aging, 349
production practices, 105, 443
protein/amino acids, 272, 307
Pseudomonas syringae pv. tabaci, 187
Q
quality assurance, 344, 388
quality assurance/cigarette production, 388
feed back and action programs, 391
instrumentation, 391, 392, 393
measurements, 389, 390
parameters measured, 393, 394, 395, 396, 397
specifications, 388, 389
quotas, 147, 164
R
radiolabeled tobacco components, 409, 411
Ralstonia solanacearum, 196
reconstituted tobacco, 20, 349, 377, 378, 379
reducing sugars, 79
respirable suspended particulates, 431
respiration, 93
Rhizoctonia solani, 192
ripeness, 126, 128
roasting, 22
root knot nematodes, 39, 216
Rotylenchulus spp., 220
S
Sclerotinia sclerotiorum, 195
Scrobipalpa heliopa, 239
seed, 11, 66
capsule, 67
light, 67
moisture requirements, 67
quality, 67
temperature, 68
seed beds, 157, 443
seed germination, 67, 167
seed storage, 69
seed types, 68
pelleted seed, 68
primed and pelleted, 68
 raw seed, 68
seedling float system, 71
clipping, 72
growing media, 71
nutrition, 71
water quality, 71
seedling production, 70
float system, 70
methyl bromide, 70
premature flowering, 70
senescence, 88, 128
sensory perception, 304
sidestream smoke, 363, 404, 406
sidestream/mainstream yield ratios, 418
smoke and tobacco constituents, 22
smoke chemistry, 398
chemical characteristics/analytical, 412
combustion process, 400
environmental tobacco smoke, 428
fate of volatile tobacco constituents, 409
formation of mainstream smoke, 402
formation of sidestream smoke, 404
 generation of specific constituents, 417
physical properties of tobacco smoke, 406
smoke composition, 415
'smoke pH', 414
smoking of cigarettes, 398
summary, 431
smoke composition, 415
smoke constituents, 23
smoke formation and composition, 353, 355, 356
smoke/fate of nicotine, 410
smokeless tobacco, 452
chemical composition, 459
manufacturing methods, 458
product types, 452
products and prevalence, 453
tobacco used, 454
smoking machines, 357, 399
smoking machines/gas phase analyzers, 396
soil insect pests, 230
cutworms, 230
false wireworms, 231
white grubs, 232
 wireworms, 232
soil pH, 171
solanesol, 277, 308, 429
spiral roots, 146
Spodoptera littoralis, 236
standard smoking method, 399
stemming, 330
stems, 336
sterols, 282, 421
stored tobacco insects, 241
cigarette beetle, 241
control, 245
tobacco moth, 242
sucker control, 148, 176
sulfur, 85
deficiency, 85
systemic suckercides, 123