Pestic. Sci.

1998, 52, 184È188

Natural Products in Agriculture—A View from

the Industry*
Martin J. Rice,” Mike Legg & Keith A. Powell
Zeneca Agrochemicals, JealottÏs Hill Research Station, Bracknell, Berks, RG42 6ET, UK
(Received 13 May 1997 ; revised version received 9 July 1997 ; accepted 26 August 1997)

Abstract : The paper discusses the use of natural products and biological control
agents in crop protection from an industrial viewpoint. The criteria which must
be satisÐed are noted. Examples are given from the genetic engineering of baculo-
viruses and proteins. The Ðnal section considers the utility of natural products as
a source of leads for conventional agrochemicals, and the screens needed.
( 1998 SCI.

Pestic. Sci., 52, 184È188, 1998

Key words : screening ; natural products ; biological control agents

1 INTRODUCTION 2.1 Does it work ?

Natural products have historically been used in numer-
ous applications in the fungal, weed and insect control
sectors of agriculture. This paper attempts to provide a
realistic overview, from an industrial perspective, of
future opportunities and difficulties attendant on the
exploration of natural products as control agents per se
or as leads from which products will be developed. For
the purposes of this paper natural products are deÐned
to include isolated compounds, biological control
agents, baculoviruses and proteins.
The efficacy of a crop-protection agent is of prime
importance. Many treatments which give an outstand-
ing performance in the laboratory or glasshouse fail to
translate this activity to Ðeld situations. There is con-
siderable variation in conditions experienced in the Ðeld
from season to season in addition to those which are a
function of geography. For a product to be a com-
mercial success, the desired e†ect must be achieved in a
robust manner in a range of climatic and agronomic
situations.

2.2 Is it safe ?

2 WHAT MAKES AN INDUSTRIAL
CROP-PROTECTION PRODUCT ?
There are a multitude of di†erent answers to this ques-
tion which can be tested by six simple criteria.
* Based on a paper presented at the meeting “Natural Pro-
ducts as a Source of Crop Protection Agents IIIÏ organised by
L. G. Copping, B. P. S. Khambay and A. Mudd on behalf of
the SCI Pesticides Group and the Royal Society of Chemistry
and held at 14/15 Belgrave Square, London, on 9 & 10
December 1996.
” To whom correspondence should be addressed.
184
( 1998 SCI. Pestic. Sci. 0031-613X/98/$17.50. Printed in Great Britain
The demands made of new products by the regulatory
bodies around the world are becoming increasingly
stringent. These demands are being driven by a com-
bination of scientiÐc and political pressures. To earn the
title of “safeÏ, a treatment is required to clear a set of
increasing higher hurdles and this trend is likely to con-
tinue as shown by the requirements for re-registration.
2.3 Is it patentable ?
The investment required to bring a new product on to
the major crop-protection markets can run at over £100
Natural products in agricultureÈa view from industry
million. In order to protect this enormous investment, a
secure patent position is essential. As prior art con-
tinues to grow, it is becoming progressively more diffi-
cult to Ðnd novelty.
2.4 Can it be made ?
In order to gain signiÐcant market share, the question
of scale-up cannot be overlooked. The extraction of a
few milligrams of a compound from a litre of fermenta-
tion broth is a straightforward procedure in the labor-
atory. To provide sufficient material for sale, massive
improvements need to be made in order to make a pro-
duction process viable. Alternatively, production by
synthesis might be technically possible and the eco-
nomics of a synthetic route compared to fermentation
have to be considered. Because of the extent of land
areas involved and the inefficiency of spraying as an
application method, agrochemicals are typically pro-
duced in amounts of hundreds or thousands of tonnes
each year. By contrast, pharmaceuticals are produced in
relatively small amounts.
2.5 Can it be sold ?
In essence, the customer is not buying the crop-
protection agent but is, in fact, purchasing the e†ect. A
product will only be successful if it addresses some need
in the market place. This might include the control of
speciÐc pests not easily dealt with by existing products,
control of a spectrum of pests occurring in a particular
crop, enabling a single rather than multiple treatment to
be used, or a spray regime that is more convenient to
the farmer, such as a single-season treatment or wider
window of application. Pharmaceuticals are more preci-
sely targeted or focused on a speciÐc disease or condi-
tion. There is a need to continue selling over a number
of years in order to recoup the investment made in
research and development. Rapid appearance of resist-
ance would, for example, severely reduce the prospects
of success.
2.6 Can money be made out of it ?
A business in the crop-protection arena has to survive
in an increasingly competitive environment. In order to
be in a position to continue to provide existing and new
products to help farmers to feed the worldÏs expanding
population into the future, sufficient proÐt needs to be
made to maintain the corporate infrastructure and fund
further research into novel areas. The shareholders also
expect a reasonable return on their investment.
185
3 BIOLOGICAL CROP-PROTECTION AGENTS
3.1 Biological control agents
The growth in biological control agents has not fulÐlled
the predictions of the mid-1980s, which were that they
would achieve a penetration of around 10% of the agro-
chemical market by the mid-1990s. The growth that has
been observed has been driven largely by consumer
preference for the source of the agent as opposed to its
e†ectiveness. Further growth is likely to be a†ected by
developments in genetic enhancement of the activity
proÐle coupled with improvements in formulation to
enhance efficacy and long-term storage stability. The
maturity of the sales of Bacillus thuringiensis Berliner
(Bt) products is likely to slow the rate of increase in
market share. The near future will see biological control
agents being used in integrated strategies and not as a
direct replacement for conventional chemical treat-
ments.
3.2 Baculoviruses
Baculoviruses o†er the greatest market opportunity of
all biological control agents. However, wild-type bacu-
loviruses face competition from broad-spectrum chemi-
cal insecticides. In this respect a major disadvantage of
wild-type baculoviruses is their slow speed of kill rela-
tive to chemicals. Genetic manipulation to enhance the
spectrum by, for example, the expression of protein-
aceous insect toxins to increase the speed of e†ect o†ers
good prospects in this area and is looking promising.
While there is optimism that the increased speed of kill
will be achieved through genetic manipulation, we still
need to see a robust e†ect in the Ðeld. Alongside efficacy
the other major technical challenge is production by fer-
mentation. Productivity will need to be dramatically
improved over current technology. Furthermore, it is
clear that for baculoviruses to compete in major
markets, production will have to be carried out on a
large scale, employing large volume (D100 000 litres)
fermenters. Culturing insect host cells on this scale is
new territory and has no guarantee of success. There
must also be a question mark over whether a baculo-
virus product which has been genetically modiÐed will
obtain regulatory approval.
3.3 Proteins
Proteins having a desired biological activity have been
regarded as leads for the production of transgenic
plants exhibiting pest-resistance traits. They also o†er
the opportunity for the discovery of novel warheads for
modiÐed baculoviruses and of novel modes of action for
186
exploitation in screening or as rational design targets.
The impact they will have on the market is unclear but
is likely to be marked. The vast majority of work in this
area has been around the Bt toxin but there are other
proteins, such as the anti-fungal proteins1 and choles-
terol oxidase2 which are beginning to change this
picture. It is currently difficult to assess how much e†ort
has been expended so far in the search but there are
undoubtedly many novel candidate proteins waiting to
be discovered.
4 SOURCES OF NOVEL NATURAL
PRODUCTS
4.1 Plants
Plants have an excellent track record in providing novel
leads for crop protection, particularly in the Ðeld of
insecticides. This can be attributed to the evolution of
secondary metabolites which address the speciÐc needs
of the host plant in protecting it from insect attack. As
products in their own right, plant natural products
present a series of challenges in both the research and
production phases. Collection of plant species for
testing may require expeditions into remote areas and
the preparation of extracts at the point of collection. If
an original small sample demonstrates interesting activ-
ity but the remaining sample is insufficient for further
progression, re-collection can be very expensive, as the
same site might have to be visited at the same time of
year in order to have the best chance of Ðnding the
same activity.
Considerations of the ownership of biodiversity have
become more prominent in recent years. While legisla-
tion or regulations regarding such ownership serve to
protect the interests of the host country, there are
dangers that the bureaucracy involved in arranging for
the collection of initial small samples of plant material
can act as a disincentive to research, leading to a loss of
potential crop-protection agents and valuable income to
the country in future years. In production, the large-
scale farming of a particular plant with the necessary
downstream processing must be compared to synthetic
approaches to the same compound, particularly if the
activity is applicable to volume, low-margin market
sectors. Plant cell culture represents an alternative
option to the above and its applicability will depend
upon the efficiency with which the compound can be
produced on a commercial scale. It is likely to be
extremely difficult to adapt this approach to the pro-
duction of commercially viable crop-protection agents.
A contrasting pharmaceutical example is the extraction
of the anti-cancer drug taxol from slow-growing yew
(T axus) species.
Martin J. Rice, Mike L egg, Keith A. Powell
4.2 Microbes
Bacteria and fungi continue to provide a rich source of
novel bioactive metabolites. The diversity of species is
immense and the production of metabolites can be
further manipulated by the use of a variety of growth
media. In principle, the preparation of extracts by fer-
mentation is possible on a sufficient scale to allow
follow-up testing and to provide material for semi-
synthetic studies of structureÈactivity relationships. In
certain cases, the high titre of a metabolite and excep-
tional potency in the Ðeld could make commercial pro-
duction by fermentation a realistic possibility. Microbes
which have demonstrated potential as biological control
agents can act as a rich source of biologically active
compounds.3
4.3 Manipulation of biochemical pathways
The production of novel compounds by the manipula-
tion of biosynthetic routes o†ers a route to “non-
naturalÏ natural products. The provision of unusual
chemicals in the growth medium can give rise to incorp-
oration via enzyme-mediated reactions to give com-
pounds which are analogues of the end product of the
pathway or intermediates which are not substrates for
subsequent transformations. A more recent approach,
exempliÐed by polyketide synthases, involves the
manipulation of the genome to produce modiÐed
enzymes which a†ord novel products.4 Such com-
pounds might be difficult to produce by other means.
The evolution of biosynthetic pathways to generate bio-
logically active compounds of use to the producer
organism is a beneÐt that is often advanced in support
of the value of natural product research. “Non-naturalÏ
natural products diverge from this origin, so it remains
to be demonstrated whether they represent a particu-
larly valuable source of novel compounds.
4.4 Leads for synthesis
Natural products have provided many challenging
targets to synthetic chemists and can result in the devel-
opment of elegant routes and new synthetic method-
ology. It is not uncommon for PhD students to devote
several years of e†ort to the construction of the complex
frameworks and functionality, with the correct stereo-
chemical orientation, of a natural product. While such
research explores the limits of what might be possible, it
is inappropriate in an industrial research programme to
expend resource on such endeavours, especially when
large numbers of analogues need to be produced to Ðne-
tune the activity to a commercial proÐle. Unless major
simpliÐcations can be made to the structure, it is
unlikely that a product will be found with sufficient
potency to justify the costs involved.
Natural products in agricultureÈa view from industry
5 SCREENING ISSUES
5.1 Compound diversity and the advent of
combinatorial chemistry
The screening of natural product extracts used to be
considered as the only feasible way of generating large
numbers of distinct and novel compounds for testing.
The advent of combinatorial chemistry, made possible
by the use of solid-phase methods of organic synthesis
and the accessibility of laboratory automation, has
mounted a challenge for the pre-eminent position of
natural products. As with many new technologies, the
claims for their capabilities are the subject of consider-
able exaggeration and it is timely to consider how com-
binatorial chemistry can complement or replace aspects
of natural-product research. Natural-product synthesis,
by classical methods, is often a multi-step procedure
involving repeated puriÐcations to remove the by-
products which result from incomplete reactions. Solid-
phase synthesis has the potential to produce highly
efficient routes to complex compounds but the develop-
ment work involved to devise the methodology is cur-
rently very slow and is unlikely to be the method of
choice for natural-product synthesis for some time to
come.
There is very little overlap between the actual com-
pounds that biosynthesis and combinatorial chemistry
will produce. It has been calculated that the number of
organic compounds with molecular weights of less than
750 which could be made theoretically using the stan-
dard rules of valency approximates to 10200.5 The sheer
size of this number becomes apparent when it is com-
pared to a conservative estimate of the number of elec-
trons in the observable universe which number a mere
1087,6 so diversity of structure is therefore not limiting.
Combinatorial chemistry has the ability to design its
compounds at the outset, not only in terms of structure,
but also in synthetic accessibility, number and ratio of
components in mixtures, polarity, diversity, novelty and
elimination of undesirable structural motifs. This gives
it the option of adapting itself to the requirements of
speciÐc high-throughput screening programmes. When
combinatorial methods Ðrst appeared, publications con-
tained claims for the synthesis of mixtures of many
thousands or indeed millions of compounds. While suc-
cessful deconvolutions have been carried out on such
mixtures to a†ord single active components, it is very
common to observe additive or synergistic e†ects and
the loss of observed activity on puriÐcation. More
recently, mixture sizes quoted in the literature have
decreased dramatically with production of single com-
pounds becoming ever more popular. The ease with
which high-throughput screens are now operated can
make it easier to screen the compounds as singles ini-
tially. The mixture size of natural product extracts is
less easy to control but methods which involve the com-
187
plete or nearly complete fractionation of the extract
prior to screening are likely to assume increasing
importance.
The removal of compounds with undesirable physical
properties is a further process for the simpliÐcation of
mixtures. Examples of methods in use include solid-
phase extraction, size-exclusion chromatography and
countercurrent chromatography. The beneÐt of sample
puriÐcation is the availability of samples with a much
higher chance of compatibility with the screens, leading
to more reliable and reproducible information on their
activity.
Rediscovery of known compounds is a perennial
challenge in natural product research. Chemical pro-
Ðling, either as an early check for the presence of spe-
ciÐc compounds or as a means of eliminating known
producer organisms, is a valuable tool in reducing
unnecessary e†ort. There has to be an element of risk-
taking in using these techniques, as an active extract
might contain a novel highly bioactive principle which
is masked by a common known metabolite. Success is
determined by what you Ðnd, not by what you avoid
missing.
5.2 Bioassays
Evaluation of the potential of an agent to produce a
commercially successful crop-protection product
requires a set of tests of ever-increasing stringency,
which in turn demands ever larger amounts of test
sample and resources to conduct tests. A number of
simple laboratory assays are in widespread use giving
Ðrst indications of activity. While such bioassays are
convenient to run, they frequently do not provide suffi-
cient discriminating information to distinguish between
weak activity and a true lead area. Examples of the limi-
tations of two of these are listed below.
5.2.1 T he wheat coleoptile assay
This bioassay is based on e†ect on growth of the test
compound on etiolated coleoptile sections from wheat
(T riticum aestivum L.) growing on an artiÐcial agar
medium, and is widely used to predict herbicide and
plant growth regulator activity.7 It is very sensitive,
registering too many false positives, and does not dis-
tinguish between herbicide and plant growth regulator
activity. Because a single species is used, there is no
information on spectrum of activity. In addition, no
symptomology is detected. This precludes the early
application of rational approaches to development of
the lead as a diagnosis of the putative mode of action is
not possible.
5.2.2 Brine shrimp assay
This bioassay is claimed to be useful for indicating
insecticidal activity, and measures the mobility of brine
188
shrimp larvae (Artemia salina L.) in a small well.8 The
aqueous environment causes lipophilic compounds to
partition preferentially into the test organism and so the
assay is biased towards detecting such compounds. The
results are sensitive to how soon after hatch the chemi-
cals are applied and it is very difficult to get sensible
results with potential insect growth regulators.
Frequently, the information on biological activity
reported in the literature is insufficient for an industrial
concern to be able to distinguish e†ectively between
those agents which have the potential to be leads or
products in their own right and those which are simply
further examples of the multitude of compounds active
only on ultra-sensitive bioassays. Many compounds
described in the literature are therefore destined to
remain as academic curiosities because their activities
have not been determined in robust tests appropriate to
agrochemical discovery.
5.3 Industry–academia interfaces
Industry has highly optimised test systems which will
detect commercially signiÐcant activity, while academia
has access to a wide variety of novel compounds from
diverse sources. There is an opportunity for collabo-
ration which is in the interests of both. Although the
chances of any one particular compound being of com-
mercial interest must been seen as low, it is important
to come to an agreement which protects the interests of
both parties in the event of a major discovery. Such
agreements are legal documents and their construction
is frequently a lengthy process as the lawyers acting for
the two parties reach a contract. As most compounds
are likely to be declared of no further interest after
initial testing by the industrial partner, it is not sensible
to enter detailed negotiations on exploitation and
royalty levels until the compound has demonstrated
that it is worthy of such attention. A simple screening
agreement will therefore contain a clause “to agree to
agree laterÏ. Should a compound make it to this stage,
any agreement should properly reÑect the relative con-
tributions of the two parties in determining levels of
interim and royalty payments. Such payments should
not only reÑect the intellectual input and time spent on
the project but also the costs involved in Ðling and
Martin J. Rice, Mike L egg, Keith A. Powell
maintaining patent applications and the share of the
risks that each party bears. For example, a compound
which becomes a product in its original form would be
expected to attract a higher royalty than one which
required the synthesis of thousands of analogues in
order to Ðnd a compound with commercially acceptable
properties.
6 CONCLUSION
In summary, compounds from natural sources have the
potential to enrich the collection of candidates for bio-
logical assessment. They bring with them their own
diversity but also a set of challenges which, if suc-
cessfully resolved, will provide the crop-protection
industry with opportunities to safeguard the supply of
food well into the next century.
REFERENCES
1. Broekaert, W. F., Terras, F. R. G., Cammue, B. P. A. &
Osborn, R. W., Plant defensins : novel antimicrobial pep-
tides as components of the host defense system. Plant
Physiol., 108 (1995) 1353È8.
2. Purcell, J. P., Greenplate, J. T., Jennings, M. G., Ryerse,
J. S., Pershing, J. C., Sims, S. R., Prinsen, D. R., Corbin, D. R.,
Tran, M., Sammons, R. D. & Stonard, R. J., Cholesterol
oxidase : a potent insecticidal protein active against boll
weevil larvae. Biochem. Biophys. Res. Commun., 196 (1993)
1406È13.
3. Faull, J. L. & Powell, K. A., Biological control agents. In
Agrochemicals from Natural Products, ed. C. R. A. Godfrey.
Marcel Dekker, New York, 1995 pp. 369È93.
4. Oliynyk, M., Brown, M., Murray, J. B., Cortes, R., Staun-
ton, J. & Leadlay, P. F., A hybrid modular polyketide syn-
thase obtained by domain swapping. Chem. Biol., 3 (1996)
833È9.
5. Czarnik, A. W., Why combinatorial chemistry, why now
(and why you should care). ChemtractsÈOrg. Chem., 8
(1995) 13È18.
6. Russell, A. (ed.), Guinness Book of Records. Guinness Super-
latives Ltd, London, 1987, p. 71.
7. Jacyno, J. M. & Cutler, H. G., Detection of herbicidal
properties : scope and limitations of the etiolated wheat
coleoptile bioassay. Plant Growth Regulator Soc. Am., 21
(1993) 15È24.
8. Blizzard, T. A., Ruby, C. L., Mrozik, H., Preiser, F. A. &
Fisher, M. H., Brine shrimp (Artemia salina) as a conve-
nient bioassay for avermectin analogs. J. Antibiot., 42
(1989) 1304È7.