Crop Improvement by Agriculture Biotechnology

Introduction to agriculture biotechnology

For about 10,000 years, farmers have been improving wild plants and animals through the
selection and breeding of desirable characteristics. This breeding has resulted in the domesticated
plants and animals that are commonly used in crop and livestock agriculture. In the twentieth
century, breeding became more sophisticated, as the traits that breeders select for include
increased yield, disease and pest resistance, drought resistance and enhanced flavor. Traits are
passed from one generation to the next through genes, which are made of DNA. All living things
including the fruits, vegetables and meat that we eat contain genes that tell cells how to function.
Recently, scientists have learned enough to begin to identify and work with the genes (DNA) that
are responsible for traits.

Agricultural biotechnology is a collection of scientific techniques used to improve plants,

animals and microorganisms. Based on an understanding of DNA, scientists have developed
solutions to increase agricultural productivity. Starting from the ability to identify genes that may
confer advantages on certain crops, and the ability to work with such characteristics very
precisely, biotechnology enhances breeders’ ability to make improvements in crops and
livestock. Biotechnology enables improvements that are not possible with traditional crossing of
related species alone. Crops can be improved by using following tools/ techniques
1. Plant tissue culture
Plant tissue culture broadly refers to the in vitro cultivation of plants, seeds, and plant parts
(tissues, organs, embryos, single cells, protoplasts) on nutrient media under closely controlled
and aseptic conditions. It includes several specialized areas, such as induction of callus and plant
regeneration, micropropagation, somatic embryogenesis, meristem culture, embryo culture,
protoplast culture, cryopreservation, and production of secondary metabolites. Tissue-culture
methods hold significant promise for creation, conservation, and utilization of genetic variability
for improvement of a wide variety of crop plants. Among these, micropropagation, embryo,
anther and protoplast culture have direct applications in crop improvement.
1.1. Micropropagation of Plants
Micropropagation of plants is one of the best and most successful examples of the commercial
application of tissue-culture technology. It entails in vitro propagation of plants from very small
plant parts (0.2-10 mm) in the laboratory, followed by their establishment in soil under
greenhouse conditions. An estimated more than 500 million plants belonging to different plant
species are now being annually produced through micropropagation in the world.
Micropropagation industry is environment-friendly and requires very little raw material in the
form of chemicals. This technology possesses the following definite advantages:
1. True-to-type plants produced, i.e., identical to donor (in vitro cloning).
2. Selected plant species can be multiplied anywhere in the world.
3. Rapid and mass multiplication (1 to 10/cycle of 2 weeks each) of elite clones/varieties
otherwise difficult to multiply through conventional methods.
4. Independent of seasonal and raw material constraints.
5. Production of disease-free plants.
6. Elite plant material for national/international exchange of germplasm, avoiding the risk of
pathogens and insects.
7. Micropropagated, field-grown plants give higher yield and exhibit better quality.
Now scores of multimillion-dollar industries around the world propagate a variety of plant
species through tissue culture, which allows environment-friendly industries to flourish. The
clean planting material can certainly improve the yield potentials of vegetatively propagated
crops like sugarcane, potato, banana, strawberry, sweet potato, cassava, and ornamental species.
It is likely that automation of multiplication systems will be commercially feasible within the
next few years. Micropropagation possesses special significance in the following areas of crop
improvement:
1. Production of high quality, disease-free, super elite planting material for further seed
production, especially in the vegetatively propagated plant species.
2. Rapid spread of new varieties of vegetatively propagated crops like sugarcane, potato, poplar,
medicinal/aromatic plants for crop diversification.
3. Mass production of ornamental plants otherwise difficult to multiply through conventional
methods for domestic and international markets.
4. Mass cloning of rootstocks in horticultural plants like citrus, peach, and apple.
5. Mass cloning of genetically superior (plus trees), cross-pollinated, and seed-propagated trees
like eucalyptus.
6. Interstate/international exchange of germplasm without the risk of pathogens and insects.
1.2. Cryopreservation and In Vitro Germplasm Storage
The aim of germplasm conservation is to ensure the ready availability of useful germplasm. In
seed-propagated crops, seed is extensively used for conservation of germplasm using
conventional methods. However, in vegetatively propagated species where conventional storage
techniques are used, it is very difficult to store germplasm for long durations. The conservation
of plant parts in vitro has a number of advantages over in vivo conservation, e.g., in vitro
techniques allow conservation of plant species in danger of being extinct. In vitro storage of
vegetativly propagated plants can result in great savings in storage space and time, and sterile
plants that cannot be reproduced generatively can be maintained in vitro. Complete plants have
been successfully regenerated from tissues cryopreserved at −196◦C in liquid nitrogen in several
crops for several months to years. This method is now being practically used at several national
and international germplasm banks.
Successful cryopreservation of plant shoot tips is dependent upon effective desiccation through
osmotic or physical processes. Cryoprotective treatments, which favor survival of small,
meristematic cells and young leaf cells, are most likely to produce high survival rates after
liquid-nitrogen exposure.
1.3. In Vitro Production of Secondary Metabolites
Several plant species are known to produce secondary metabolites in vivo or in vitro. These
metabolites do not perform vital physiological functions, but some act as potential predators and
attract pollinators. Further, these metabolites act as a valuable source of a vast array of chemical
compounds, including fragrances, flavors, natural sweeteners, and industrial feed stocks.
Cultured cells/organs produce a wide range of secondary products. Mainly three approaches have
been followed: (i) the rapid growth of cell suspension cultures in large volumes, (ii) elicitation of
plant cells cultures by various biotic and abiotic factors, and (iii) growing hairy root cultures in
vitro. There are several advantages of the cell-culture systems over the conventional cultivation
for production of secondary metabolites, e.g., 1) independence from various environmental
factors; 2) any cell of the plant could be multiplied to yield specific metabolite; and 3) a culture
of cells may prove suitable in cases where plants are difficult or expensive to grow in the field
because of their long life cycles. Since the 1970s when the possibility of producing useful
secondary products in plant cell cultures was first recognized, considerable progress has been
made, and a number of plant species have been found to produce secondary products, such as
shikonin, diosgenin, caffeine, glutathione, capsaicin, and anthraquinone. Large-scale production
of such compounds (molecular farming) is becoming increasingly popular with the industry
where some physical and chemical conditions for growth and product formation have been
optimized.
2. Molecular Pharming
An additional goal for the development of transgenic plants is the use of living systems for the
production of metabolites at the industrial scale, e.g., specialty chemicals, antibodies,
pharmaceuticals, edible vaccines, etc. The cell’s metabolic networks that evolved in nature are
not optimized for industrial production of these metabolites. So, the performance of metabolic
pathways is manipulated so that metabolites are over-produced. The introduction of heterologous
genes and regulatory elements in the living systems is commonly called ‘metabolic engineering,’
There are a number of reports dealing with the production of specialty chemicals,
biopharmaceuticals, and edible vaccines that can be stored and distributed as seeds, tubers, or
fruits. Solulin is a recombinant, soluble derivative of human thrombomodulin. Edible vaccines
against gastrointestinal tract infections have been developed in banana.
3. Gene cloning
With the availability of a variety of cloning vectors, useful genes can now be isolated, cloned,
and characterized. The discovery of the reverse transcriptase enzyme made it possible to isolate
genes for specific proteins or genes having tissue-specific expression. Nowadays, PCR cloning
has become routine especially for those genes whose base sequences are known. Several genes,
particularly for disease resistance, have been isolated and cloned from genomic or cDNA
libraries using heterologus or synthetric probes. Using such genes, suitable gene constructs
carrying marker gene(s), promoter, and terminator sequences are developed for plant
transformation.
4. Molecular markers
Development and utilization of molecular markers to detect differences in the DNA of individual
plants has many applications of value to crop improvement. These differences are known as
molecular markers because they are often associated with specific genes. Several types of
molecular markers that have been developed and are being used in plants include: restriction
fragment-length polymorphism (RFLP), amplified fragment-length polymorphism (AFLP),
randomly amplified polymorphic DNA (RAPD), sequence-tagged sites (STS), expressed
sequence tags (ESTs), simple sequence repeats (SSRs), and single nucleotide polymorphisms
(SNPs). Such markers, closely linked to genes of interest, can be used to select indirectly for the
desirable allele, which represents the simplest form of marker-assisted selection (MAS) and is
now being exploited to accelerate backcross breeding and pyramid several desirable alleles.
Using molecular markers, genotypes can be distinguished at plant, tissue, or even cellular level.
DNA markers are now extensively being used for gene mapping/tagging. Likewise, molecular
markers are also extensively used to probe the level of genetic diversity among different varieties
and related species. The applications of such evaluations are many, including DNA profiling,
fingerprinting, efficiently managing genetic resources, and facilitating introgression of
chromosomal segments from alien species. In addition, markers and comparative mapping of
various species have been very valuable in enhancing our understanding of genome structure and
function, and have allowed the isolation of genes of interest via map-based cloning.
5. Crop improvement by genetic engineering
5.1 herbicide-resistant transgenic plants.
5.2 Engineering for Insect Resistance
5.3. Engineering for Disease Resistance
a). Virus resistance
b). Fungal resistance
c). Bacterial resistance
5.4. Engineering for Abiotic Stress Tolerance
5.5. Engineering for Improved Nutritional Quality
There is now a growing interest in improving the nutritional quality of food crops. Genes cloned
from plants and microbes are being introduced into crop plants to enhance their nutrional value.
For instance, introduction of provitamin A and β carotene genes have resulted in the production
of ‘golden rice’. Likewise, high protein ‘phaseolin’ and AmA1 genes have been introduced to
hetereologous systems such as tobacco and potato. Intoduction of AmA1 gene into potato has
improved its yield, protein content, and quality. Vitamin producing transgenic plants have also
been developed and emphasis is being laid on multigene engineering. The main objective of
these crops is to add value to agri-foods.