Plant Biotechnology

Plant biotechnology is a set of techniques used to adapt plants for specific needs or
opportunities. Situations that combine multiple needs and opportunities are common. For
example, a single crop may be required to provide sustainable food and healthful nutrition,
protection of the environment, and opportunities for jobs and income. Finding or developing
suitable plants is typically a highly complex challenge. Plant biotechnologies that assist in
developing new varieties and traits include genetics and genomics, marker-assisted selection
(MAS), and transgenic (genetic engineered) crops.

Plant Biotechnology is being projected as a contributor to augment food supply both

quantitatively and qualitatively. The advancements in understanding of plant physiological
processes, phytochemistry, biosynthetic pathways, molecular biology, genomics, proteomics
and metabolic regulation have provided impetus to the progress. Plants are being modified for
high photosynthetic efficiency; adoption to extreme environments such as frost; drought, salinity,
alkalinity; to fight disease causing microbes and damages from insect pests; and for increased
responsiveness to agrochemicals for increasing crop yields.

Attainment of Food Security

Meeting the Food Security of the world population has been a big social concern and
responsibility. It is projected that the world population will reach 9 billion by 2050 (UN Report
data, 2004). The adoption of high yielding wheat and rice in the 1970s, coupled to fertiliser
usage, ushered in the green revolution. However, despite the best efforts of agricultural and
plant scientists, the increase in yield for most of the food crops has nearly reached a plateau.
Increase in food productivity is possible with a broadening of agricultural base and adoption of
biotechnology for plant improvement. There may be a need to shift total dependence on
popular crops such as rice, wheat, corn and soybean to several locally cultivated traditional
varieties. For example, supplementation with crops like cassava in Africa; minor millets such as
ragi, jowar, pearl millet etc in India, will be of relevance to utilise biodiversity for enhanced food
production in a region specific manner. The biotechnological interventions such as selection of
superior lines through marker assisted selection (MAS) for resistance to pests and diseases and
adaptability to various stress conditions for crop yield enhancements is warranted (Utomo et al.,
2009).
DNA markers are useful in improving plant characteristics by conventional breeding methods
adopting MAS. Development of Quantitative Trait Loci (QTLs) mapping for crop plants provides
DNA-trait associations (Collard and Mackill, 2008). Adoption of these technologies for
improvement of traditional crops has great relevance to improve food and nutritional security.
The advancements in genomics and proteomics, combining the tools of molecular biology and
bioinformatics, have already enhanced our capabilities to understand the molecular basis of
plant characteristics. These are high impact areas of relevance to agriculture for enhancing plant
productivity through the gene revolution.
Erosion of the gene pool is a matter of concern. Ex situ and in situ gene banks supported
through germplasm conservation by cryopreservation, seed bank and tissue culture
methodologies are promising. Such efforts would facilitate international exchange of germplasm
for plant improvement. Utilisation of wasted lands for enhancement of agricultural base requires
development of crops, which are suitable for extreme environment conditions such as drought,
floods, frost, salinity, alkalinity, etc.
Valuable additions to agrimaterials, global marketing strategies, efficient distribution systems
and development of food products which are nutritionally superior, affordable and convenient,
needs attention. Extensions of shelf life of staple grains, fruits and vegetables have been
attempted through biotechnological interventions. Prevention of post harvest losses will
promote induced farming since it will enhance returns to growers and farmers.

Nutritional Security
Though the aspect of nutritional security also has an angle of affordability, use of local varieties
and germplasm combining regional practices and modern biotechnological interventions will
help mitigate the problem to a great extent by expanding the agricultural base. The excessive
dependence on cereals as staple foods is often faced with the problem of lysine deficiency.
Already, plant biotechnological methods for increasing lysine in cereals and methionine in
legumes have been achieved. The discovery of high-lysine opaque 2 mutants in maize (Mertz,
1997) led to the search for similar mutants in other cereals. However, such mutants were
susceptible to diseases and exhibited low yields. Genetic separation of a high-lysine trait from
undesirable traits which impair productivity was successfully done to develop quality protein
maize (Gallili et al., 2002) which is an exemplary biotechnological effort worth emulating.

Essential amino acids viz., lysine, methionine, isoleucine and threonine are biosynthesised from a
common precursor, aspartate. Generally, amino acid production is limited by feed back
inhibition of their own pathway. The regulatory gene for dihydropicolinate synthase from a
bacterium which is insensitive to feedback inhibition was adopted in maize for over-production
of lysine (Mazur et al., 1999). It was later found that understanding lysine catabolism is
important for overproduction and accumulation. Only when the plants have all desirable
characters in addition to high lysine can the same be used.
The prevalence of Vitamin A deficiency, protein malnutrition and micronutrient deficiencies is
continuing unabated, affecting nearly 2 billion people world wide. Such nutritional deficiencies
can be met through the use of traditional foods which are an abundant source of quality
protein, vitamins and other micronutrient levels etc. Efforts in this direction to provide newer
sources of these nutritionally important foods and food ingredients are gaining attention. The
use of micro algae (Chlorella, Scenedesmus, Spirulina) for food applications as a source of
protein and micronutrients has been well recognised (Ravishankar et al., 2006).
Biotechnological production of these algal forms in large scale open bioreactors in race-way
ponds has been a successful industrial endeavour. Algae based products for food applications
are unique, and novel processes are being developed for valuable food ingredients (table 2).

Genetic engineering of plants was initially aimed at enhancement of productivity, mainly

focusing on tolerance to agrochemicals and herbicides (Gressel, 2002). These modifications,
which may be termed as the first generation of Genetically Modified (GM) plants, have been
questioned from an environmental safety angle. However, genetic modifications conferring
resistance to insect pests, by cloning genes for Bacillus thuringiensis toxin (Nayak et al., 1997 ) or
resistance to fungal pathogens by cloning Chitinase, glucanase genes (Chye and Zho, 2005),
have a bearing on plant productivity with less usage of pesticides and fungicides. This
technology is likely to gain support due to reduced agrochemical usage benefiting the farmers
and providing eco-friendly alternatives. General lack of popularity of GM technology, from the
consumer point of view, could be due to inadequate efforts towards a qualitative improvement
of foods. There is increasing need to address the utility concerns of all stake holders e.g.
consumer, farmers and food processors.
Food Additives through Biotechnology
The turn of the last century witnessed changes in public preference from synthetic to natural
ingredients such as natural colours, sweeteners, flavourants and taste enhancers. Several
candidate molecules have been identified for application as food additives (table 7).
The aspects of their production through biotechnological processes and the establishment of
safety, efficacy, absorption levels and metabolism will continue to be the subject of interest.
Improvement in Processing Characteristics

Improvement in processing characteristics of agri-materials with a focus on product

development is of interest to the food industry. Biotechnological intervention for value addition to
processing is being pursued in several crops (table 3).
Food quality improvement will be of benefit to consumers (table 4).
However, there is a need to educate all the stake holders to realise the benefits of the modern
plant biotechnologies for the societal advantage and to enhance acceptability.

Improvement in Nutraceutical Value

Nutraceuticals, functional foods or health foods are gaining popularity. The food based
approach to health maintenance has long been realised and is evident from the Chinese system
of herbal medicine and also the Indian system of herbal remedies, namely Ayurveda.
Prospecting for the functional attributes of foods and validating traditional knowledge with
modern analytical methods have lead to newer understanding of nutritional and health benefits
of food ingredients.
Quality improvement in foods is facilitated by enhancement of basic nutrients and
nutraceuticals. The present day functional foods aim to cater to life style and life cycle based
approaches. Thus, foods containing specific attributes such as immuno-enhancement, cardiac
protection, diabetes foods and disease prevention are becoming popular (table 5).
This has culminated in the production of designer foods (Siber, 1997) with functional ingredients
(table 6).
Personalised foods - based on physiological status and genetic make up are likely to become
popular with the advancement of nutrigenomics (Muntoni and Muntoni, 2008).

Metabolic Engineering

Scope of expansion of the functional food sector is dependent on the research on the
identification of potential biological sources of biomolecules; sustainable production
methodologies; metabolic engineering of pathways by way of suppression of undesirable steps,
or over expression of regulatory steps; and establishment of safety, generating appropriate
toxicological data for regulatory considerations. The engineering of rice (golden rice) by cloning
beta carotene genes (Beyer et al., 2002 ), enhancement of prebiotic polysaccharides like
fructans in sugar beet (Weyens et al., 2004) and the manipulation of anthocyanin pathway
genes are a few other examples of pathway engineering for value addition to agri-foods. The
powerful tools such as gene pyramiding, antisense gene technology, RNAi, microRNA and
tissue specific expression systems will facilitate the achievement of the desired traits in a
targeted manner.

Improvement in Plant Oil

Over 100 million tonnes of vegetable oil are produced for food purposes annually of which 70
million tonnes are derived only from a few crops such as soybean, canola, palm and sunflower.
The increase in their productivity has been achieved through traditional breeding methods.
However, value addition aimed at the improvement of the quality of the edible oil will possibly be
achieved by pathway engineering. Such an endeavour has led to the development of γ linolenic
acid (GLA) rich oil seed crop by designing the fatty acid pathway. Isolation of the Δ6 desaturase
gene from borage and its introduction to soya has enriched GLA production which may provide
nutritionally superior oil. GLA enhances the immune response, useful for heart ailments and for
brain development (Fan and Chapkin, 1998). The enhancement of oxidative stabilisation of
edible oil, as measured by either the active oxygen methods (AOM) or Oxidative Stability Index
(OSI), is an important parameter determining stability to oxidation and polymerisation.
Generally, bleached and deodorised vegetable oil is useful for frying once with AOM of 15-25
hours, whereas for food industry usage it needs to be stabilised much more. High AOM of 150
hours can be reached with high oleic soybean oil (Kinney, 1988). High oleic acid production in
soybean of up to 85% of the seed oil (Mazur et al., 1999) has been achieved by inserting an
antisense gene which inhibits the enzymatic step that includes a second double bond at the Δ12
position into oleic acid (18:1Δa). These transgenics are reported to be highly superior without
alteration of other characteristics. Furthermore, soybean is richer in γ-tocopherol than other
plant oils. Since the gene for tocopherol formation has been cloned it is a matter of time before
tocopherol-rich stable oil is produced by combining modifications in tocopherol metabolism and
fatty acid desaturation.
Moreover, superior oleic acid-rich oil is also likely to be available through the technology of
pathway engineering.

Pesticide-Resistant Crops

In the past, farmers have incurred significant losses due to the use of pesticides that affect both
crops and weeds. Biotechnology has led to the engineering of plants that are resistant to
pesticides. This allows farmers to selectively kill weeds without harming their crop. A famous
example is the Roundup-Ready tech introduced by Monsato.
The tech was first introduced in genetically modified soy beans, making them resistant to the
herbicide glyphosate. The herbicide can be applied in copious amounts to eliminate other plants
on a field other than the Roundup-Ready plants. Selective elimination of weeds saves farmers’
valuable time as compared to traditional methods of weeding.
Pest-Resistant Crops

For many years, a microbe known as Bacillus thuringiensis (Bt) has been used to dust crops by
producing toxic proteins against pests. One of such toxic proteins used for dusting crops is the
European corn borer. Scientists have come up with a way to eliminate the use of Bt by
introducing pest resistant crops. These are known as Bt crops as the gene that’s introduced in
the crop was originally identified in Bacillus thuringiensis. Examples if pest resistant crops today
are Bt maize, potato, and corn. This toxic protein is only harmful to pests, but is safe for
humans. It has saved farmers from dealing with expensive pest infestations in crops.

Role of Food Biotechnology in Food Processing

Fermentation

Breweries are synthesized through the process of fermentation. Different yeast strains are used
to make breweries at commercial level. Genetic engineering has enabled us to make light wine.
Yeast is genetically modified through foreign gene encoding glucoamylase. During process of
fermentation yeast expresses glucoamylase that convert starch into glucose [3].

Yeast strains used for wine synthesis are capable of malolactic fermentation. Wine synthesis
consists of two steps: 1) Primary fermentation results in conversion of glucose
into alcohol using yeast. 2) Secondary fermentation uses bacteria and its product is lactic acid
and this causes the rise in level of acidity. To overcome this problem different strategies are
used which are costly. This problem was solved through insertion of malolactic gene
(Lactobacillus delbrueckii) in industrial yeast strain. This gene lowers the malate conversion
hence lowering acidity level of wine [4,5].

Enzymes

Enzymes are used in production and processing of food items specifically produced at industrial
level. From second last decade of twentieth century, food processing companies are
using enzymes that are produced through genetically modified organisms (European food
information council 2015). These enzymes comprises of proteases and carbohydrases. Genes
for these enzymes have been cloned so as to get higher production in less time period. These
enzymes are used for making cheese, curd and flavoring food items. Major percentage of these
enzymes is used in food industry as in US more than 50% of proteases and carbohydrases are
used in food industry. These enzymes include rennin and α-amylase [3].

Following are some genetically modified enzymes used in food industry:

• Catalase used in mayonnaise production and it removes hydrogen peroxide.

• Chymosin useful in cheese production as it coagulates milk.

• Glucose oxidase is used in baking as it stabilizes the dough.

• α-amylase converts starch into maltose and used in baking for sweetness.

• Protease used for meat tenderization process, baking and dairy products [6].

α-amylase

This enzyme is used in the production of high fructose corn syrup (nutritive sweetener). This
enzyme provides continuous process of three steps providing higher yield. Through purification
this yield can be increased up to 90%. In 1986, Grant devised a system to produce α- amylase
through genetic engineering using Bacillus subtilus as a host. Plasmid named pCPC720 was
use as vector. In same year FDA approved this system of genetic engineering to synthesize α-
amylase to be used at industrial level [7].

Rennin (Chymosin)

Rennin enzyme is an active component of substance rennet used in dairy industry. It is a

protease enzyme used for the production of curd and cheese. This hydrolysis the peptide bond
of casein proteins of milk, hence denaturing these proteins results in curd formation [8].

Previously this enzyme was extracted from stomach of calves and used to curdle milk. But
through this conventional method, lower quantity was obtained. But now bacteria (Escherichia
coli ) and fungi (Aspergillus niger ) are genetically engineered to produce rennin at commercial
level [3].
Shell Life

Many juicy fruits possess short shell life. For example tomato is used all over the world. In order
to be shipped, tomatoes should be picked at mature green stage. After picking, these are
subjected to ethylene for ripening. Higher temperatures cause early ripening while lower
temperature destroys its taste.

A Californian company named Calgene genetically engineered tomato to sort out this problem.
They developed a tomato named Flavr Savr tomato. An enzyme named polygalacturonase
breaks down pectin causing ripening and softening. Scientists genetically modified tomatoes to
reduce amount of this enzyme. They used antisense RNA for this purpose [2]. Low amount of
this enzyme results in lower breakdown of pectin and cell wall resulting in firmer tomatoes.
These firmer Flavr Savr tomatoes possess longer shell life and hence support shipping [9].

Biotechnology: Improving Food Nutrition

Every food item does not contain all essential components. That’s why every food article is not
possessing perfect nutrition. For example rice is used as staple food in many countries of world.
But being devoid of vitamin A, it’s not a perfect staple food. Use of Biotechnological techniques
has solved these problems through introduction of foreign vitamin A gene [10].

Proteins and essential amino acids: More than half of worldwide protein production is
attained from plants but plant proteins lack some essential amino acids like lysine and sulphur
containing amino acids [10]. Corn is genetically modified and it expresses proteins produced by
soil bacteria Bacillus thuringiensis [11]. To overcome the deficiency of essential amino acids,
different biotechnological molecular processes are used and given below (Table 1).

  Name of transgenic Molecular pathway for Enhanced Essential amino Foreign genes
plant modification acids incorporated

1 Tobacco Synthetic gene approach Overall amino acids Asp1

Manipulation of gene Sulphur containing amino acid Gene encoding 2S

2 sunflower seed
expression (MET) albumin

Manipulation of homologous
3 Potato Mostly amino acids AMA1
protein

Table 1: Plants and food modified with amino acid genes, source of gene of interest and
pathway of modification.

Vitamins and minerals: These are a compulsory food component that’s why to avoid their
deficiency, transgenic technology is used. Rice is one of the foods used as staple food in many
countries of world. But being deficient in Vitamin A, rice is not a perfect staple food. The first
provitamin rich transgenic rice was produced by incorporating crtI gene and psy gene from
bacteria and daffodils [10]. Variety of provitamin rich rice can eliminate malnutrition and
blindness from developing countries and third world. [11]. Scientists are working on introduction
of other vitamins and macronutrients (iron, zinc etc.) genes in vitamin deficient food articles
Iron: Iron is one the most important minerals required for a healthy body. The countries which
use rice as a staple food are more vulnerable to iron deficiency because rice is deficient of iron
[12]. To resolve this problem, rice is transformed with a foreign gene encoding iron containing
gene named ferritin. Transformed rice contains double content of rice as compared to non-
transformed rice

Carbohydrates and lipids: Carbohydrates, lipids can be modified in transgenic plants. In late

20th century, amylopectin rich potatoes and lauric acid rich canola oil was produced through
agricultural biotechnology [10]. Potatoes have been genetically modified by inserting a gene
from bacteria that encode enzyme involved in starch biosynthesis pathway. These GM potatoes
contain 30-60 % more starch

Use of Biotechnology to Improve Yield

Milk is of the food item used all over the world due to its nutritional value. Bovine Somatotropin
is a hormone released by pituitary gland. It raises the milk production. Previously this hormone
was extracted from brain of slaughtered calves. But that results in low quantity. Scientists
inserted gene encoding bovine Somatotropin in Escherichia coli . Now this hormone is obtained
in higher quantity. This hormone results in 10-12% rise in milk production [12,14].

By the year 2050, population of world will become nine billion. So more yield will be requiring on
same land. Biotechnology is potentially best technology to fight against problem of food yield

Africa has highest level of poverty and hunger. This hunger and malnutrition results in diseases
like kwashiorkor and rickets which in turn causes a lot of deaths. Biotechnology possesses
highest potential to make Africa get rid of hunger, starvation, malnutrition and diseases. It can
raise health standard and lower mortality rate. Three African countries: Burkina Faso, South
Africa and Egypt have already been benefitted through adaptation of biotechnological cultivation
methods. For example 0.1 million farmers of Burkina Faso raised yield of cotton by 126%
through use of GM food technology.

Adoption of GM food technology required system for commercial release of GMO products,
tests for allergenicity, digestivity and toxicity of GM food. In this area USA and European Union
should help Africa. Many African countries lack biosafety system. African should make biosafety
law making and approval their priority so as to make this system adopted easily.

Biotechnology: Enhancing Taste

Biotechnology has allowed scientists to produce fruits with better taste. GM foods with better
taste include seedless watermelon, tomato, eggplant, pepper and cherries etc. Elimination of
seed from these food articles resulted in more soluble sugar content enhancing sweetness
[11]. Fermentation pathways are modified using biotechnology to add aroma in win [3].