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Agricultural Biotechnology: current status and future prospects

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

Agricultural Biotechnology: current status and future prospects

*Vikas
Kumar and 1Lishamol Poulose
*Dept. of Silviculture & Agroforestry, Kerala Agricultural University, Thrissur- 680656
1
Muslim Educational Society. College Marampally, Aluva, kerala-683107
*Corresponding author Email: vksumar49@gmail.com

Abstract: Agricultural biotechnology represent a broad range of technologies used in food and agriculture for
the genetic improvement of plant varieties and animal populations, characterisation and conservation of genetic
resources, diagnosis of plant or animal diseases and other purposes. Discussions about agricultural
biotechnology have been dominated by the continuing controversy surrounding genetic modification and its
resulting products, genetically modified organisms (GMOs). The polarised debate has led to non-GMO
biotechnologies being overshadowed, often hindering their development and application. Modern biotechnology
represents unique applications of science that can be used for the betterment of society through development of
crops with improved nutritional quality, resistance to pests and diseases, and reduced cost of production.

Key words: Agriculture, Biotechnology, GMOs, development of crops, nutritional quality, plant parasitic
nematode control, livestock, aquaculture, forestry and ABDC-10.

Kumar et al., 2015a). But most frequently

Introduction:
today the term is used to refer to
knowledge about the natural processes of
The world population now is 6.7 billion
DNA replication, breakage, ligation, and
and is predicted to reach 9 billion by 2050
repair that has made possible a deeper
(Von Braun, 2010). Such a rapid growing
understanding of the mechanics of cell
population has tremendously increased the
biology and the hereditary process
challenge for food security. Obviously, it
(McCouch, 2001). Biotechnology provides
is impossible for traditional agriculture to
important tools for the sustainable
ensure the food security, while plant
development of agriculture, fisheries and
biotechnology offers considerable
forestry and can be of significant help in
potential to realize this goal (Fedoroff,
meeting an indispensable part in the rise of
2010). The term “biotechnology” has been
human civilization. It indeed often
used to refer to many biological processes
considered as one of the fields of scientific
that produce useful products, including
research in which the most rapid advances
some quite ancient ones such as
have been made in recent years (Vikas
fermentation in beer, wine and cheese
Kumar et al., 2015a). Along with the
(Coombs, 1992; Zaid et al., 1999, Vikas
increasing commercial success of
Vol. II, New Academic Publishers, New Delhi (ISBN: 978-8186772751)
“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

application of biotechnology, a widespread development of climate resilient farming

debate focusing on the ecological, human and livelihood systems, although it is
health and socio-economic effects of rapidly being lost.
biotechnology is taking place at national
The strong controversy about the subject
and international levels (Rimal et al.,
of agricultural biotechnology relates to one
2003).
single biotechnology, genetic
Agricultural biotechnology is a collection modification, and its resulting products,
of several scientific techniques such as genetically modified organisms (GMOs).
genetic engineering, molecular breeding, Indeed, the term biotechnology is often
molecular diagnostics, tissue culture, and used synonymously for genetic
vaccines in order to improve crop and modification (hence the need for a
livestock productivity and the definition earlier). The debate about the
marketability of derived products. On the advantages and disadvantages, real or
other hand, it a collection of scientific perceived, of GMOs began over a decade
techniques used to improve plants, animals ago and it still continues today without
and microorganisms. Based on an showing significant signs of abating. The
understanding of DNA, scientists have positions of different parties regarding
developed solutions to increase GMOs have often become firmly
agricultural productivity. Starting from the entrenched in what has been called a
ability to identify genes that may confer ‘global war of rhetoric’ (Stone, 2002).
advantages on certain crops, and the ability
Applications of agricultural
to work with such characteristics very
biotechnology
precisely, biotechnology enhances
breeders’ ability to make improvements in
The primary tools used in agricultural
crops and livestock. They are also used to
biotechnology are defined below
characterise and conserve agricultural
biodiversity and Professor M.S. i. Genetic engineering inserts
Swaminathan, in his keynote address to fragments of DNA into
ABDC-10 (FAO, 2011a), noted that chromosomes of cells and then
biodiversity has been the feedstock for uses tissue culture to regenerate
sustainable food and health security and the cells into a whole organism
that it can play a similar role in the with a different genetic

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

composition from the original crossing two plants that would not
cells. This is also known as rDNA normally produce offspring.
technology; it produces transgenic iv. Somatic hybridization removes the
organisms. cell walls of cells from different
ii. Tissue culture manipulates cells, organisms and induces the direct
anthers, pollen grains, or other mixing of DNA from the treated
tissues; so they live for extended cells, which are then regenerated
periods under laboratory into whole organisms through
conditions or become whole, tissue culture.
living, growing organisms; v. Marker-aided genetic analysis
genetically engineered cells may studies DNA sequences to
be converted into genetically identify genes, QTLs (quantitative
engineered organisms through trait loci), and other molecular
tissue culture. Embryo rescue markers and to associate them
places embryos containing with organismal functions, i.e.,
transferred genes into tissue gene identification.
culture to complete their vi. Marker-aided selection is the
development into whole identification and inheritance
organisms (Fig 1). tracing of previously identified
DNA fragments through a series
of generations.
vii. Molecular diagnostics are methods
to detect genes or gene products
that are very precise and specific.
Molecular diagnostics are used in
agriculture to more accurately
Fig 1: Plant improvement through diagnose crop/livestock diseases
tissue culture technology (James, 2003; (http://www.geo-
iii. Embryo rescue is often used to pie.cornell.edu).
facilitate “wide crossing” by viii. Genomics analyzes whole genomes
producing whole plants from of species together with other
embryos that are the result of biological data about the species
to understand what DNA confers

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)
what traits in the organisms.
Similarly, proteomics analyses the
proteins in a tissue to identify the
gene expression in that tissue to
understand the specific function
of proteins encoded by particular
genes. Both, along with
metabolomics (metabolites) and
phenomics (phenotypes), are
subcategories of bioinformatics.
ix. Vaccines are used in livestock and
humans. They may be cheaper,
better and/or safer than traditional
vaccines. They are also stable at
room temperature, and do not
need refrigerated storage; this is
an important advantage for
smallholders in tropical countries.
Some are new vaccines, which
offer protection for the first time
against some infectious illnesses.
For example, in the Philippines,
biotechnology has been used to
develop an improved vaccine to
protect cattle and water buffalo
against hemorrhagic septicemia, a
leading cause of death for both
species (Ives et al., 2001;
http://www.nysaes.cornell.edu).
Artificial insemination (AI) and
multiple ovulation embryo
transfer (MOET): have already
had a major impact on livestock
Vol. II, New Academic Publishers, New Delhi
improvement programmes in
developed countries and many
developing countries because they
speed up the process of genetic
improvement, reduce the risk of
disease transmission and expand
the number of animals that can be
bred from a superior parent - the
male in the case of AI and the
female in the case of MOET.
They also increase the incentives
for private research in animal
breeding and significantly expand
the market for improved parent
stock.
x. Biofortication involves the process
of development of nutritionally
enhanced crop plants using
biotechnology. Using techniques
of genetic linkage mapping, genes
responsible for enhanced
nutritional levels are transferred
to crops through conventional
breeding or genetic engineering.
Example, to increase the protein
content in maize and iron in rice.
xi. Cryopreservation refers to storage
of living cells, tissues or organs at
ultra-low temperatures, usually
that of liquid nitrogen (-196°C)
(Xu et al., 2011). When stored at
such a low temperature, cellular
divisions and metabolic processes
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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

of the living cells arrest and 1. Crops:

therefore, plant materials can be
Crop biotechnologies have developed
conserved for an indefinite period
incrementally over the past century, but
of time, while maximally
progress has accelerated greatly over the
maintaining their genetic stability
last two decades, leading to many
(Benson, 2008). Furthermore,
important scientific achievements and
cryopreservation occupies little
impressive technological advances.
space, eliminates contamination
Genetic modification has had limited but
and demands only maintenance.
real success in modifying a few simple
Therefore, cryopreservation has
input traits in a small number of
been considered an ideal means
commercial commodity crops, adopted
for safe and long-term storage of
also in some developing countries.
plant germplasm (Benson, 2008;
However, the major breeding and crop
Reed, 2008). Successful
management applications to date have
cryopreservations have been
come from non-GMO (genetically
reported for transgenic cell
modified organisms) biotechnologies
suspensions that expressed human
encompassing the full range of agronomic
serum albumin (HAS) in
traits and practices relevant to developing
Nicotiana tabacum (Schmale et
countries farmers. In 2005, genetically
al., 2011), Escherichia coli heat
engineered varieties of maize (corn),
labile enterotoxin (LT) protein in
cotton, canola, and soybean were widely
N. tabacum (Van Eck and Keen,
planted in North America and Asia while
2009) and hCTlA4Ig in O. sativa
there was minimal use of such varieties in
(Cho et al., 2007).
Europe and Africa. Most transgenics have

Here, a brief overview is provided of the been engineered to confer a single plant

ways that agricultural biotechnologies are trait (Dunwell, 2000; Chilton, 2005; Bajaj

being used to increase productivity and and Mohanty, 2005; Halpin, 2005; Sharma

conserve natural resources in the crop, et al., 2005), but multiple trait varieties

plant parasitic nematode control, livestock, comprised 20% of the total transgenic

aquaculture, forestry, agro-industry and crops in 2005 (James, 2004). The global

ABDC-10 prepared by FAO (Chapters 1–5 area of transgenic crops first exceeded 1

respectively in FAO, 2011a).. million hectares in 1996; over the next

four years the area increased to over 40
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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)
million hectares, reaching 90 million
hectares by 2005 (James,
Transgenic rice was grown commercially
for the first time in 2005, in Iran, and is
being widely field tested in China.
Although not directly contributing to food
consumption, transgenic cotton adds to
income and hence food security and has
rapidly spread in the United States, China,
India, and elsewhere, covering over 5
million hectares. Only a handful of other
transgenics, including papaya and squash,
are commercially grown (Sankula et al.,
2005). Transgenic soybean, cotton, and
maize adoption rates have been extremely
rapid by historic standards, similar to those
of the green revolution in
(Dalrymple, 1975) and of hybrid corn in
the United States (Griliches, 1957).
Biotechnology also offers important tools
for the diagnosis of plant diseases of both
viral and bacterial origin, and immuno-
diagnostic techniques as well as DNA-
based methods are commercially applied
for this purpose in some developing
countries (Miller et al.,
Additionally, biotechnologies such
molecular markers, cryopreservation and
in vitro slow growth storage
extensively used for the characterisation
and conservation of plant genetic resources
in developing countries (FAO, 2011b).
Vol. II, New Academic Publishers, New Delhi
Although still tiny compared with
2004). synthetic pesticides, the market for
biopesticides is increasing worldwide, with
developing countries responsible for under
20% of global biopesticide production
(Thakore, 2006). For example,
biopesticides formulated with the spores of
the fungus Metarhizium anisopliae var.
acridum have been used successfully to
control migratory locusts in countries such
as Timor-Leste and Tanzania (FAO,
2011a).
2. Plant parasitic nematode control:
Research on biotechnological approaches
to nematode control aims either to exploit
Asia natural resistance present in gene pools of
crop species and their relatives or to
employ synthetic forms of resistance, such
as those based on disruption of feeding
cells, expression of specific proteins or
peptides, on gene silencing (RNAi) or on
delivery of toxic compounds to the
invading nematode (Table 1). To exploit
natural variation for resistance, large-scale
screening of germplasm is often employed,
2009).
together with molecular markers and/or
as
positional cloning to identify resistance
(R) genes or metabolites that confer
are
resistance to particular nematodes in a
wide range of germplasm of crop plants
and their wild relatives. Identified sources
of resistance are then introgressed into the
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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

desired germplasm. In contrast, transgenic finding host roots, reducing the

approaches to nematode control exploit effectiveness of migration through host
knowledge of nematode–host interactions tissues, reducing successful establishment
and can be directed to targeting the in host cells or reducing feeding ability
nematode, including disorientating the and fecundity of nematodes on a
infective stages to prevent them from susceptible or tolerant host (Table 1).

Table 1: Biotechnology-Based Strategies for Nematode Control (Fosu-Nyarko and Jones,

2015).

Sl. Target for Considerations Status/Example

No. Control
1. Major or minor The introgression and Marker-assisted breeding for
natural resistance combination of natural nematode resistance has become
genes. resistance genes, for example routine in many breeding
from related or wild species, programs, although effective
has been the mainstay of resistance genes are not available
resistance breeding strategies. for all crops.
2. Nematode Disruption of sensory Peptide(s) that inhibit reception of
migration in the functions. gradients by amphids RNAi
rhizosphere and disruption of amphid
root entry. proteins/function.
3. Migration in the Wall-degrading enzymes may RNAi downregulation of
root. be required for migration, e.g. nematode expression of cell wall-
Endoparasites Positional degrading enzymes.
gradients in roots detected for
Inhibition of sensing gradients in
migration to the required site
roots..
in the root.
4. Avoiding host Effectors that enable RNAi down regulation of
defences nematodes to evade or expression of effectors involved in
neutralize host defences avoiding host defences.
5. Disruption of Effectors enable sedentary RNAi downregulation of
feeding site endoparasites to induce giant expression of key effector(s)
formation or cells and syncytia. Disrupt

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

function. feeding site formation, required for feeding site formation.

triggered by nematode-
Nematode responsive promoter(s)
responsive promoter(s).
linked to ‘cell death’ gene, e.g.
barnase.
6. Disrupting vital Disrupt expression of genes RNAi down regulation of
genes vital for the nematode life expression of vital nematode genes
cycle.
7. Overexpression Many genes in nematode Over expression of some host
of host genes feeding cells are up-or genes with altered expression in
with modified downregulated. nematode feeding sites reduces
expression in nematode parasitism.
feeding cells.
8. Modify genes for New approaches for genome New technologies not necessarily
host plant editing now available. regarded as genetic modification,
susceptibility to more acceptable in some
nematodes. jurisdictions.
9. Delivery of toxic Make use of basic work on Use bioinformatics filters to
compounds to the nematode effectors and genes identify new targets for chemical
nematodes. vital for their survival: these control. Design new nematicides
can define new targets for to these targets.
control.
10. Develop new There is a need to develop A series of new nematicides are
nematicides and new more environmentally now available commercially, based
modes of friendly forms of chemical on biological and chemical
delivery; new control and delivery, and new control, separately or in
biological control forms of biological control. combination, e.g. using delivery
agents. by drip irrigation or seed coating.
financial security. They contribute 40% of
3. Livestock: the global value of agricultural output

Livestock contribute directly to livelihoods (FAO, 2009), and this proportion is

worldwide, providing not only food, but expected to increase. Animal

also non-food products, draught power and biotechnology has proceeded in two

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

directions: the production of animals for increasingly used to allow early diagnosis
meat or milk and the creation of animals of diseases, although their use is mainly
that produce biomedically useful proteins restricted to the laboratories of research
in their blood or milk. The latter is a institutions and larger governmental
highly specialized application directed at diagnostic laboratories. Vaccination has
human health and is limited to a few been widely used as a cost-effective
companies (Faber et al., 2003). measure to control infectious diseases, as
exemplified by the case of rinderpest, an
The lack of a system for the identification
infectious viral disease of cattle, buffalo,
of superior animals limits, along with lack
yak and numerous wildlife species that has
of technical capacity, the use of more
caused devastating effects throughout
advanced technologies, such as embryo
history (John Ruane and Sonnino, 2010).
transfer or marker assisted selection
(Nimbkar and Kandasamy, 2011). 4. Aquaculture:
Transgenic research on farm animals is not
Compared with livestock and crop
widely conducted, and progress is
production, aquaculture is a novel
relatively slow (Faber et al., 2003; Hansel,
production system in many developing and
2003) for several reasons. Molecular
developed countries, and it has made less
biotechnologies in the area of animal
use of conventional technologies, such as
reproduction and breeding in developing
traditional genetic selection, and
countries have generally been limited to
biotechnologies to increase production
genetic characterisation studies, usually
than these other sectors (John Ruane and
through international cooperation. Farm
Sonnino, 2010). These include the
animals have much longer reproductive
manipulation of sex in fish using hormonal
cycles than plants; so seeing the result of
treatment to generate single sex
any breeding innovation takes much
populations, for example in tilapia (Cnaani
longer. Techniques for superovulation,
and Levavi-Sivan, 2009). Hormonal
ovum recovery, in vitro fertilization,
treatment has also been used successfully
nuclear transfer, cloning, and embryo
to control the timing of reproduction in
transfer have low rates of success;
fish and shellfish, for example in salmon
therefore, applying genetic engineering to
and trout farming in Chile.
animals is inefficient (Hernandez et al.,
2005). PCR-based diagnostics are

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

For better understanding of the population monodon and Penaeus vannameii is

structure of aquaculture is therefore of practised in all countries producing
paramount importance. The use of commercial shrimp (FAO, 2011a).
molecular markers and the principles of Biotechnology tools have been used
population genetics have proved very widely in their development, where they
effective for assessing the actual levels of have facilitated antigen discovery;
genetic variability within single construction of new candidate vaccines;
populations and for measuring the extent and the assessment of vaccine efficacy,
of differentiation between populations. mode of action and host response (Kurath,
Some of the main ways in which 2008). Some biotechnologies are being
molecular marker data have been applied used to address these areas, including
for conservation decision-making in fish bioremediation for the degradation of
populations include characterising the hazardous wastes (Chávez-Crooker and
genetic structure of the populations being Obreque-Contreras, 2010) and use of DNA
harvested; detecting changes/falls in based methodologies for the early
population size; and estimating the detection of toxin-producing algae.
effective population size (Primmer, 2006).
5. Forestry:
Disease outbreaks are a serious constraint
The world has slightly less than 4 billion
to aquaculture development. Better
hectares of forests, covering 31% of the
management of intensive systems is
world’s land area (FAO, 2010b). 30% of
needed, and biotechnologies are assisting
the world’s forests are primarily used for
in this task. Immunoassay and DNA based
production of wood and non-wood
diagnostic methods are currently applied
products. Only 7% of forests in the world
for pathogen diagnosis in developing
are in plantations, with the balance found
countries. For example, twenty years ago
in natural or semi-natural, largely
in the shrimp sector, which is the most
unmanaged and undomesticated forest
valuable aquaculture commodity sector in
stands. Forest trees have unquestionably
the world; there was hardly any accurate
entered the genomic era. Biotechnological
molecular-based pathogen detection
tools viz., Transgenic technology, RNA
system available in any part of the world.
interference, Functional genomics, Marker
Now, PCR detection of viruses of
assisted selection, QTL and tissue culture
broodstock and postlarvae in both Penaeus
etc. has paved road for successful

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)
exploitation and integration of scientific
fields with an increased sense of urgency
for delivery of cutting edge research in tree
biotechnology, both in academia and
industry (Vikas Kumar et al., 2015a).
Today, findings are available to guide
operational forest management plans,
including in developing countries, but only
for a very limited number of the hundreds
of tree species that are managed in
naturally regenerated tropical forests. This
area of forest biotechnology continues to
expand (Muchugi et al., 2008; Kindt et al.,
2009).
Forestry has enormously benefited from
development and implementation of
improved silvicultural, forest management
practices and breeding techniques have
contributed significantly to the
improvement of forest tree species in the
past, and will continue to have a
substantial impact on the genetic gain and
productivity of economically important
tree species by providing better germplasm
and improved management practices for
plantation forests. Although good progress
has been made in breeding trees for altered
xylem-fibre lengths and lignin content,
which is valuable to the paper and pulp
industries (Turnbull, 1999; Heilman,
1999), much less progress has been made
in improving timber quality, precisely
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because wood formation is so poorly
understood (Lev-Yadun and Sederoff,
2000; Plomion et al., 2001). It is probably
one of the most complex phenomena
facing plant biologists today, with perhaps
40000 genes being involved (Lorenz and
Dean, 2002), so without biotechnological
tools to gain a better understanding of the
process, markers for wood quality traits
will remain a distant prospect. Traditional
breeding methods are often constrained by
the long reproductive cycles of most tree
species and the difficulty in achieving
significant improvements to complex traits
such as wood properties, disease and pest
control, and tolerance to abiotic stresses.
The state of food and agriculture reported
that “biotechnology is more than genetic
engineering” (FAO, 2004). In fact, 81% of
all biotechnology activities in forestry over
the past ten years were not related to
genetic modification (Wheeler, 2004).
Genetic engineering has the potential to
boost global wood production in many
ways (Sutton, 1999; Sedjo, 2001).
Applications currently under consideration
for plantation forests include resistance to
biodegradable herbicides, altered lignin
properties for reduced downstream
processing costs or improved burning,
resistance to selected pests, altered
reproductive mechanisms for faster
breeding or genetic containment,
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phytoremediation of polluted sites, and the widely planted as cuttings in family

production of novel chemicals or or clonal plantations (Ritchie, 1991;
pharmaceuticals (Strauss, 1999; Strauss et Ahuja and Libby, 1993; Assis et al.,
al. 2001; Yanchuck, 2001). Only limited 2004; Menzies and Aimers-Halliday,
studies are available on the role of 2004).
biotechnology in plantation forests ii. Micropropagation: The vegetative
(Fenning and Gershanzon, 2002), propagation of plants has been
economic benefits resulting from the practiced for centuries and many
introduction of forest biotechnology improvements in conventional
(Sedjo, 2001), and ecological issues methods have been made over the
associated with the deployment of years. Recently, the tissue culture
genetically modified forest tree species technique i.e micropropagation has
(Van Frankenhuyzen and Beardmore, expanded their scope and potential
2004). on commercial scale.
i. Propagation: Chinese fir Micropropagation is suitable for the
(Cunninghamia lanceolata) has been rapid and large-scale clonal
propagated by cuttings for clonal multiplication of elite germplasm.
forestry in China for more than 800 The technique has been referred as
years (Li and Ritchie, 1999) and micropropagation because the size of
Japanese cedar (Cryptomeria the tissue in culture is very minute as
japonica) has been propagated compared to conventional vegetative
clonally by cuttings in Japan for cutting or any other plant part. The
plantations since the beginning of meristem explant used for
the fifteenth century (Toda, 1974). micropropagation is about 0.1-0.5
Some tree species are easier than mm size having only one or two leaf
others to propagate by cuttings. Easy primordial (Vikas Kumar et al.,
to root hardwood species, such as 2015b). Morel and Martin (1952) for
poplars (Populus spp.), willows the first time demonstrated that virus
(Salix spp.) and some eucalypt free plants can be obtained by
(Eucalyptus) species, and conifer culturing shoot meristems. Later on
species, such as spruces (Larix spp.), with the discovery of the hormonal
redwood (Sequoia sempervirens), control of organogenesis by Shoog
and some pines (Pinus spp.), are and Miller (1957) and finding of

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

most commonly used T/C media by subcultured after 2-3 weeks, on

Murashigue and Skoog (1962), the shoot multiplication (Fig 2)
scope of micropropagation was medium. The medium is designed
further extended to vast range of in such a way to avoid the
plant species, including fruit and formation of callus, which is
plantation crops. With the undesirable for true to type
advancement in science and multiplication of plants. Thus the
technology, micro propagation careful use of auxins like NAA,
technique has also been standardized 2,4-D and cytokinins like BAP,
for many plants, and it, is now Kinetin is done in culture medium.
widely used for multiplication of It is well-established fact that
many horticultural plants. cytokinins enhance shoot
Stages involved in Micropropagation multiplication.

There are four main stages involved in III. Rooting of shoots: The in vitro

micropropagation of plants, such as regenerated shoots are rooted in the

explants establishment, shoot proliferation, medium containing auxins like

rooting of shoots, hardening and transfer to NAA, IBA. The rooting can also be

soil/field. induced (Fig 2) when in vitro

shoots are exposed to stress
I. Explant establishment: The conditions. The rooting should
establishment of explant depends also be preferably without
on several factors such as the formation of callus, thus avoiding
source of explant/ genotype, type somaclonal variants.
of explant such as leaf, root, stem IV. Hardening and transfer to soil/
from mature or immature plants/ field: The in vitro plantlets thus
seedlings, explant sterilization, the obtained are hardened/ aclimatized
in vitro culture conditions such as before transfer to the field. The
culture media, composition, hardening is necessary as the
temperature, humidity, light etc. Tissue culture derived plants grow
The explants showing growth are under high humidity conditions,
considered established (Fig 2). have open stomata, lower
II. Shoot multiplication: The epicuticular wax, thus leading to
established explants are increased transpiration losses and

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resulting in mortality of plants (Fig 2).

Figure 2: Micropropagation techniques A. Selection of explant plant of Banana, B.

Inoculated explants in culture, C. Callus induction, D. & E. Shoot induction, F. & G. Root
induction, H. Shoot proliferation, I. Transplanting of plant, J. Hardening process under shade
net.

Limitations of micropropagation IV. Pathogens once appeared in the

system, they also multiply at a very
I. The facilities required are very
faster rate in a short time.
costly.
V. Plants having high levels of
II. Technical skill is required to carry
phenols (mango. date palm,
out different micro propagation
coconut etc.), usually do not
procedures.
respond to micropropagation
III. High cost of nutrient media and
techniques
sterilizing agents

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)
VI. Lack of systems for marketing/
delivering tissue culture products
VII. Establishment of laboratory-raised
plants in the field is a very difficult
task
The disadvantages of micropropagation
method
a. The need to set up a small
laboratory, which can be
expensive for a small nursery
b. The need for a well trained
technician and other skilled staff
c. The need for more manual labour
to transfer the seedling to
different size pots during its
growing period.
d. It takes longer to get the plantlets
ready for planting
e. More susceptible to pest and
disease on site and adverse
conditions.
The following advantages of
micropropagation techniques
 Year around production of plants
irrespective of seasonal constraints.
 Small space is required to maintain
and multiply large number of
plants.
 Small tissue is required as an
explant, hence saves the scion
wood to a great extent.
Vol. II, New Academic Publishers, New Delhi
 Speedy international exchange of
germplasm, requiring minimum
quarantine checks is possible.
 Micropropagated plants exhibit
vigorous growth, and higher yields.
 Micropropagated plants are usually
free from viruses
 Micro propagation is highly
beneficial in dioecious fruit plant
species (date palm and papaya),
where large-scale production of
female plants is possible.
 It helps in reducing the breeding
cycle.
 Production of homozygous plants
is possible under in vitro conditions
 It is highly beneficial in plants in
which vegetative propagation is not
possible or the propagation rate is
very slow (papaya and date palm)
 In vitro systems have the potential
for long-term transportation or
shipment of propagation material.
Problems encountered during
micropropagation
The success of micropropagation in
several instances is hampered by the
following problems (Vikas Kumar et al.,
2015b).
a. Microbial contamination:
Bacterial/fungal contaminations in
the cultures do not allow the
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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

propagules to grow. This problem from callus instead of direct shoot

can be overcome by growing donor induction and proliferation.
plants in growth chambers, Moreover, the plants regenerated
systemic fungicide spray prior to through adventitious meristems as
explant removal, effective compared to axillary meristem are
sterilization of explants, susceptible to mutations, as it is
performing inoculations in laminar derived from either a single cell or
air flow cabinets fitted with HEPA a small group of cells. Thus leads
filters (0.2m) and using sterilized to variation in regenerated plants.
surgical instruments. Fumigation The variation due to callusing, can
of inoculation room using dilute be overcome by addition of growth
formaldehyde solution also helps to substances which inhibit callusing
minimize this problem. such as triiodobenzoic acid
b. Browning of cultures: The (TIBA), phloroglucinol and
cultured explants of certain plant phloridzin and also by reduction of
species secrete phenolic substances inorganic salt concentration in the
into the medium, which cause culture medium.
browning due to oxidation of d. Loss of plants due to
phenols and formation of quinones, transplantation shock: Tissue
the toxins which effect the growth culture regenerated plants have
of cultured explants. The use of anormal leaf morphology, poor
antioxidants such as activated photosynthetic efficiency,
charcoal (1-2%), citric acid or malfunctioning of stomata (open),
ascorbic acid (50-100 mg/l) and reduced epicuticular waxes and
polyvinylpyrrolidone (PVP), thus are amenable to
polyvinylpolypyrrolidone (PVPP) transplantation shock. Hardening
in the culture medium helps to of such plants is thus must before
check the browning. transplantation under field
c. Variability in T/C regenerated conditions. Conservation of
plants: Variability is highly moisture by creating high humidity
undesirable in the micropropagated around the plants, partial
plants. It may occur due to defoliation and application of anti-
callusing and regeneration of plants

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)
transpirants are useful
hardening of in vitro raised plants.
iii. Genetic Transformation: The first
successful transformation in trees
was achieved by Fillatti et al., 1987.
Since then progress has been slow
but steady and has witnessed many
new inventions and techniques over
past decade, which have
reviewed extensively (Merkle and
Dean, 2000; Pena and Seguin, 2001;
Herschbach and Kopriva,
Diouf, 2003; Gallardo et al., 2003;
Gartland et al., 2003). The main
theme to attempt genetic
transformation in trees is
improvement of productivity and
quality.
iv. Transgenesis: A wide diversity of
sources of transgenes and regulatory
elements, and intended traits, have
been tested, including expression of
reporter genes; insect, disease, and
herbicide resistance; modified wood
properties; modified flowering and
fertility; and modified growth rate
and stature (Viswanath et al., 2011).
The major obstacles to efficient
production of transgenic trees are: (i)
difficulties in plant regeneration
from Agrobacterium-infected
particle-bombarded explants;
incomplete development beyond the
Vol. II, New Academic Publishers, New Delhi
for in vitro stage of rooted plants for
establishing field trials; and (iii)
transgene instability during the long
lifespan of forest trees, including
transgene silencing and somaclonal
variation (Harfouche et al., 2011).
Once transgenesis is performed at
the cell level, in vitro culture
been techniques can be used to regenerate
the entire tree (Vikas Kumar et al.,
2015b).
2002; v. Abiotic stress resistance: The use
of known abiotic-stress-associated
genes from other species to enhance
tolerance in forest trees has been
the limited. However, recent studies in
genomics, transcriptomics and
proteomics in several forest tree
species, as well as release of the
draft Eucalyptus grandis genomic
sequence (www.eucagen.org), have
provided new tools for improving
abiotic stress tolerance in trees
(Harfouche et al., 2011). Over
expression of a pepper ERF/AP2
transcription factor, CaPF1, in
eastern white pine resulted in a
significant increase in tolerance to
drought; freezing and salt stress
(Tang et al., 2007). The increased
or tolerance was associated with
(ii) polyamine biosynthesis. Moreover,
over expression of the choline
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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)
oxidase (codA) gene from
Arthrobacter globiformis resulted in
increased tolerance to NaCl in
several lines of Eucalyptus globules
(Yu et al., 2009).
vi. Biotic stress resistance: The use of
genetic engineering to improve tree
resistance to insects and microbial
pests has been the subject of
investigation in several laboratories.
Genetically engineered insect
resistance can be environmentally
beneficial because of the reduced
need for synthetic insecticides.
Hybrid triploid poplars [(Populus
Tomentosa X P. bolleana) X P.
tomentosa] transformed with
cowpea trypsin inhibitor gene (CpTI)
exhibited resistance to three
defoliating insects: forest tent
caterpillar (Malacosoma disstria),
gypsy moth (Lymantria dispar) and
willow moth (Stilpnotia candida)
(Zhang et al., 2005). Forest trees
play host to a wide range of fungal,
bacterial and viral pathogens. Trees
engineered for disease resistance can
provide both environmental and
commercial benefits. Enhanced
disease resistance has been achieved
using a variety of genes derived from
plants and microorganisms, with
varying degrees of success. For
Vol. II, New Academic Publishers, New Delhi
example, Chinese white poplar (P.
tomentosa) expressing a chitinase
gene from Beauveria bassiana
(Bbchit1) exhibited increased
resistance to a pathogenic fungus
(Cytospora chrysosperma) (Jia et al.,
2010). Testing for disease resistance
in a natural setting is imperative and
multi-year field trials will be needed
to verify the durability of resistance
against ever-evolving pathogen
populations (Vikas Kumar et al.,
2015a).
vii. Tissue culture: Tissue culture or
genetically modified trees will be
substantial managers with time to
a evaluate many of the issues being
faced in agriculture; the economic
realities of relatively long
generations will continue to be a
major challenge to investors in
biotechnology in forest trees. It
appears that genetic modification
will therefore become a reality only
for particularly novel and valuable
traits in short-rotation species in
intensively managed plantations.
Based on the Indian government’s
aim to enlarge the total area covered
by forests to 23% by 2010 and to
33% by 2020, biotechnology is
going to play a central role to tackle
specific challenges. Trees are long
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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)
living organisms and often
surrounded by their wild relatives,
which causes a risk of gene flow
from transgenic stands into natural
populations. Research is still needed
to investigate the problems related to
transgenic trees, e.g. concerning all
aspects of biosafety including efforts
to prevent the escape of transgenes
into natural populations. Approval
for the commercial use of transgenic
trees and their easy vegetative
propagation by cuttings (e.g. in
poplar, eucalyptus, casuarinas and
salix) can cause a rapid distribution
of transgenic plant material in the
near future (Vikas Kumar et al.,
2015b).
6. Agro-industry:
Agro-industries provide a means of
converting raw agricultural materials into
value added products while generating
income and employment and contributing
to overall economic development.
Biotechnology as applied to food
processing uses fermentation and
microbial inoculants to enhance properties
such as the taste, aroma, shelf-life, texture
and nutritional value of foods (e.g. FAO,
2011a; Liu et al., 2011). Hybridisation is
also used for the improvement of yeast
strains. Genetic modification is widely
employed in R&D for strain improvement
Vol. II, New Academic Publishers, New Delhi
(Olempska-Beer et al., 2006).
Biotechnological developments have led to
the widespread availability of methods of
identification that are more rapid and less
costly than those based on conventional
techniques. PCR-based and enzyme-linked
immune absorbent assay methods are now
applied for the detection of major food-
borne pathogens (Velusamya et al., 2010).
7. ABDC-10:
FAO organised the international technical
conference on Agricultural
Biotechnologies in Developing Countries
(ABDC-10) that took place in Guadalajara
from 1 to 4 March 2010, hosted by the
Government of Mexico. A major objective
of the conference was to take stock of the
application of biotechnologies across the
different food and agricultural sectors in
developing countries, in order to learn
from the past and to identify options for
the future to face the challenges of food
insecurity, climate change and natural
resource degradation.
Current status
Biotechnology has been proven better for
the improvement of agricultural
biotechnology. Genetically biotechnogy
plants have also reduced the need of
fertilizers thus minimized the pesticide
pollution to rivers and costal water
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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

resources. The techniques like sound understanding of the science and

development of DNA vaccines and technologies involved, the public is wary
genetically altered bacteria and other of genetically altered foods and concerned
transformations of life through about the inability to control biological
biotechnology have been pursued for the agents once they are released into the
sake of the social benefits which environment. However, the harms that may
biotechnologist’s promises. Cheaper and result from such interventions typically do
more effective medicines are possible not remain confined to those interests or
when produced through biological rather the individuals responsible for introducing
than chemical means. But appropriate them, but instead may propagate
balance of environmental and health throughout the environment and affect the
concerns against economic benefits is general public. A gene that protects a food
essential for the use of biotechnological crop from certain pests benefits the farmer
applications. and the seed company directly, but should
One of the first modifications through that gene cross into a noxious species, it
genetic engineering in microorganisms may well create problems for the general
was done in bacteria that have the ability public.
to digest oil spilled in the oceans.
Future prospects
Bioremediation and in general, the
improvement of the environment have In recent decades, a significant number of
been the primary aims of a great deal of investigations were performed on
biotechnological research. In the marine agricultural biotechnology. Most of those
context, much of the scientific work being known were discovered, increase the
done is aimed at ameliorating the effects productivity of crops, primarily by
on food species and marine ecosystems of reducing the costs of production by
overdevelopment, pollution, and loss of decreasing the needs for inputs of
breeding habitats. While biotechnological pesticides, give better rotations to conserve
methods promise a variety of important natural resources, improved the
social and environmental benefits, public agricultural production, by developing
response, especially to the release of varieties having resistance against biotech
genetically modified species into the and abiotech stresses. Production of
environment, has been mixed (Ahmad et diseases free plants with improved
al., 2015). Though not always based on a nutritional value has proven the role of

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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)
biotechnology in agriculture sector.
Nevertheless, the affordability and
accessibility of next generation sequencing
technology is bound to transform the field
of applied genomics, there by accelerating
the ability to decipher the genetic factors
behind agricultural important traits. Many
tree species engineed for expression of a
variety of traits are already under extensive
cultivation in many parts of the word. The
status of biotechnology in India is very
encouraging and the future back on with
many opportunities. All modern
biotechnologies require large research and
development investments. The allocation
of funds, through either private or public
agencies, needs to achieve a balance
between building scientific capabilities and
knowledge and supporting more applied,
well proven forestry technologies. From a
genetic perspective, concerns that
biotechnology is "unnatural" ignore the
dynamic changes in the genetic code that
occur within and across species genomes
through modification of transposable genes
or elements by virus vectors and through
mutation. Tissue culture or genetically
modified trees will be substantial
managers with time to evaluate many of
the issues being faced in agriculture, the
economic realities of relatively long
generations will continue to be a major
challenge to investors in biotechnology in
Vol. II, New Academic Publishers, New Delhi
forest trees. It appears that genetic
modification will therefore become a
reality only for particularly novel and
valuable traits in short-rotation species in
intensively managed plantations. However,
while those in the forefront of any
technology will promote its potential
benefits, in the end it will be the economic
and regulatory systems of governing
bodies at the national and global levels that
must evaluate the technology's relevance
and appropriateness.
Conclusion
The aim of this review was to present the
state of the art of the applications of
agricultural biotechnology are being used
to increase the productivity of crops,
primarily by reducing the costs of
production by decreasing the needs for
inputs of pesticides, mostly in crops grown
in temperate zones. The application of
agricultural biotechnology can improve the
quality of life by developing new strains of
plants that give higher yields with fewer
inputs, can be grown in a wider range of
environments, give better rotations to
conserve natural resources, provide more
nutritious harvested products that keep
much longer in storage and transport, and
continue low cost food supplies to
consumers. Biotechnologies could provide
us with useful tools in sectors such as
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“Biodiversity, Conservation and Sustainable Development” (Issues & Approaches)

agriculture, horticulture, forestry, fisheries, S.A. and Amjad, M. 2015. Ethical

food production and industry. Crop Issues of Biotechnology, Possible
production will have to cope with rapidly Risks and Their Management.
increasing demand while ensuring Journal of Biology, Agriculture and
environmental sustainability. Preservation Healthcare. 5(11): 49-54.
of natural resources and the need to
Ahuja, M.R. and Libby, W.J. (editors).
support the livelihoods of farmers and
1993. Clonal forestry II:
rural populations around the world are
conservation and application.
major concerns. In order to achieve the
Springer-Verlag, Berlin and
best solutions, we must consider all the
Heidelberg, Germany.
alternatives for addressing these challenges
using independent and scientifically sound Assis,T.F., Fett-Neto, A.G. and Alfenas,
methods. These alternatives include A.C. 2004. Current techniques and
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the world is serving as a source of Plantation forest biotechnology for
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agricultural biotechnology.
Bajaj, S. and Mohanty, A. 2005. Recent
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