CHAPTER 13

BIOTECHNOLOGY AND ITS APPLICATIONS

APPLICATIONS OF BIOTECHNOLOGY
❖ Therapeutics
❖ Diagnostics
❖ Genetically modified crops for agriculture
❖ Processed food
❖ Bioremediation
❖ Waste treatment
❖ Energy production
THREE CRITICAL RESEARCH AREAS OF BIOTECHNOLOGY
❖ Providing the best catalyst in the form of improved organism usually a microbe or
pure enzyme.
❖ Creating optimal conditions through engineering for a catalyst to act.
❖ Downstream processing technologies to purify the protein/ organic compound.
BIOTECHNOLOGICAL APPLICATIONS IN AGRICULTURE
THREE OPTIONS FOR INCREASING FOOD PRODUCTION ARE:-
❖ Agro- chemical based agriculture
❖ Organic agriculture
❖ Genetically engineered crop based agriculture
The green revolution succeeded in increasing food supply because of:
• Use of improved crop varieties
• Use of agro- chemicals (fertilisers and pesticides)
• Use of better management practices.
Agrochemicals are expensive for the farmers in developing countries and also have
harmful effects on environment. Also it was not enough to feed the human population.
One of the alternative to this is the use of genetically modified crops in agriculture.
TISSUE CULTURE
➢ It is an in vitro technique of regeneration of whole plant from any part of a plant by
growing it on culture medium under sterile conditions.

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 ➢ The part of the plant taken out for tissue culture is called explant.
➢ The basic principle behind tissue culture is totipotency.
➢ Totipotency is the capacity to generate whole plant from any cell or explant.
COMPONENTS OF NUTRIENT MEDIUM

• A carbon source – Sucrose

• Inorganic salt
• Vitamins
• Amino acids
• Growth hormones
ADVANTAGES OF TISSUE CULTURE
• A large number of plants can be grown in a short time. This method of producing
thousands of plants through tissue culture is called micro- propagation.
• Plants produced through tissue culture are genetically identical to the original plant
from which they are grown. So they are called somaclones.
• Seed less plants can be multiplied through tissue culture.
• Disease – free plants can be obtained by tissue culture by using meristem as explant.
• Tissue culture is used to develop somatic hybrids by somatic hybridisation.
MERISTEM CULTURE
❖ Healthy plants can be recovered from diseased plants by this method.
❖ Meristem (apical and axillary) is the only virus- free part of a virus infected plant.
❖ By removing the meristem and growing it in vitro, virus – free plants can be obtained.
SOMATIC HYBRIDISATION
The process of fusion of somatic cells obtained from different varieties or species of plant on
a suitable nutrient medium in vitro to develop a somatic hybrid is called somatic
hybridisation.
STEPS OF SOMATIC HYBRIDISATION
➢ Single cells from selected plants are isolated.
➢ The cell walls of cells are digested by enzymes like pectinase and cellulase to expose
the naked protoplasts.
➢ Naked protoplats surrounded only by plasma membranes are isolated.
➢ The isolated protoplasts are fused to obtain hybrid protoplasts under sterile
condition, in special nutrient media.

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 ➢ The hybrid protoplasts regenerate cell wall and divides to form a new plant.
Eg:-Pomato is a somatic hybrid between potato and tomato that belong to two different
genera
GENETICALLY MODIFIED ORGANISMS (GMO)
Plants, bacteria, fungi and animals whose genes have been altered by manipulation are
called Genetically Modified Organisms (GMO).
ADVANTAGES OF GM PLANTS
➢ Made crops more tolerant to abiotic stresses like cold, drought, salt, heat,etc.
➢ Reduced reliance on chemical pesticides.
➢ Helped to reduce post- harvest losses.
➢ Increased efficiency of mineral usage by plants.
➢ Enhanced nutritional value of food. Eg:- Vitamin ‘A’ enriched rice (golden rice)
(i) PRODUCTION OF PEST- RESISTANT PLANTS
• One of the important application of biotechnology in agriculture is the production of
pest – resistant plants.
• It decreases the amount of pesitcides used.
Eg:- Bt cotton, Bt corn, rice, tomato, potato, soyabean,etc.
Bt Cotton
Cotton plant having Bt toxin gene (cry gene) is called Bt cotton.
• Bacillus thuringiensis is a soil bacterium which has a gene called cry gene (Bt toxin
gene)
• This bacterium produces a crystal protein (cry protein) or Bt toxin protein.
• This protein can kill certain insects like lepidopterans (tobacco budworm, army
worm), coleopterans (beetles) and dipterans (flies, mosquitoes).
• During the particular phase of its growth, B. thuringiensis produce protein crystals
known as cry proteins.
• Cry proteins are produced in an inactive form as protoxins and do not kill the
bacterium.
• Cry proteins on ingestion by the insects are transformed into soluble active form in
the alkaline pH of the gut.
• The activated proteins bind to the epithelial cells of the mid gut creating pores.
• This causes cell swelling and lysis which results in the death of the insects.

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TYPES OF Bt TOXIN GENES
Cry I Ac - Controls the cotton boll worms
Cry II Ab - Controls the cotton boll worms
Cry I Ab - Controls corn borer
This genes were isolated from B. thuringiensis and incorporated in many plants like
cotton, corn, rice, tomato, etc.
(ii) PEST RESISTANT PLANTS
A nematode Meloidegyne incognitia infects the roots of tobacco plants which reduce
the production of tobacco. Resistance has been introduced in crop plants by using
RNA interference (RNAi)
RNA INTERFERENCE
❖ This is a method which involves silencing of a specific mRNA due to a
compementory double stranded RNA (dsRNA) molecule that binds to and prevents
translation of the mRNA.
❖ The source of this complementary RNA could be an infection by viruses having
RNA genome or mobile genetic element known as transposons.
The steps for making tobacco plants resistant to nematodes:
➢ Nematode specific genes were introduced into the host cells by using Agrobacterium
as a vector.
➢ The DNA was introduced into the host cell in such a way that , it produce both sense
and anti sense RNA in the host cells.
➢ Since both RNA’s were complementory, they form a double stranded RNA which
initiated the RNAi process.
➢ The initiation of the RNAi process does not allow the specific mRNA of the
nematode to express itself, thus silencing it.
Thus the transgenic plant got itself protected from the parasite.
BIOTECHNOLOGICAL APPLICATION IN MEDICINE
GENETICALLY ENGINEERED INSULIN
Earlier insulin used for diabetes was extracted from pancreas of slaughtered cattle
and pigs. Insulin from an animal source caused some patients to develop allergy or
other type of reactions to the foreign protein.
Structure of insulin
• Insulin contains two short polypeptide chains – chain A and chainB linked by
disulphide bridges.

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 • In mammals, insulin is synthesised as a pro- hormone. It contains an extra stretch
called C peptide.
• C peptide is absent in mature insulin and is removed during maturation into insulin.
• In 1983, Eli Lilly, an American company prepared two DNA sequences
corresponding to A and B chains of human insulin and introduce them in plasmids
of E.coli to produce insulin chains.
• Chains A and B were produced separately, extracted and combined by creating
disulfide bonds to form human insulin.
GENE THERAPY
Gene therapy is a collection of different methods which involves correction of a
defect, diagnosed in a child or in an embryo.The normal gene is inserted into the cell
or tissue of the child or embryo to treat hereditary diseases.
The first gene therapy was given in 1990 to a four year old girl with Adenosine de
aminase (ADA) deficiency caused due to the deletion of a normal gene.
• ADA is the crucial enzyme for the immune system to function.
• The patient has non- functioning T- lymphocytes.
• So they cannot produce immune responses against invading pathogens.
• SCID (Severe combined immunodeficiency) is caused due to deletion of the gene for
the ADA synthesis.
STEPS OF GENE THERAPY FOR ADA DEFICIENCY
• Lymphocytes from the blood of the patient are grown in a culture outside the body.
• The functional ADA, cDNA is introduced into these lymphocytes through retroviral
vector.
• These lymphocytes are then introduced into the body of the patient.
Since lymphocytes have a short life span, these have to be replaced periodically.
• The therapy becomes completely curative when the genes producing ADA are isolated from the
bone marrow cells and are introduced into the cells of the early embryo.
• In some children, ADA deficiency can be cured by bone marrow transplantation
and enzyme replacement therapy. But these methods are not fully curative.
MOLECULAR DIAGNOSIS
An early diagnosis of a disease is very important for the effective treatment of the
disease. Through conventional methods of diagnosis, early detection of a disease is
not possible as the pathogen is present in low concentration in these body fluids.
SOME TECHNIQUES USED FOR EARLY DIAGNOSIS ARE:

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 ❖ PCR
❖ Recombinant DNA technology
❖ ELISA (Enzyme Linked Immuno- sorbent Assay)
APPLICATIONS OF PCR
➢ HIV infection in suspected AIDS patient.
➢ Mutations in genes of cancer patient.
➢ To identify many genetic disorders.
➢ To amplify DNA for DNA finger printing.
RECOMBINANT DNA TECHNOLOGY
• A single stranded DNA or RNA tagged with a radioactive molecule is called a
probe.
• In this method, a probe is allowed to hybridise to its complementary DNA in the
clone of cells.
• The cells are then detected by autoradiography.
• The cell with mutated gene will not be observed on the photographic film because
the probe will not have complementary DNA strand to the mutated DNA strand of
the gene.
ELISA
• ELISA test is an immunochemical clinical test commonly used to detect antibodies
and antigens in a sample by antigen- antibody interaction.
• Infection by pathogen can be detected by the presence of antigens or by detecting the
antibodies synthesised against the pathogen.
TRANSGENIC ANIMALS
Animals whose DNA is manipulated to possess and express an extra gene are known as
transgenic animals.
REASONS FOR DEVELOPING TRANSGENIC ANIMALS
❖ Study of normal physiology and development.
❖ Study of disease.
❖ Biological products.
❖ Vaccine safety.
❖ Chemical safety testing.
1. Normal physiology and development

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 • Useful to study gene regulation, their effect on the normal functions of the
body and its development.
For eg:- study of complex factors involved in growth such as insulin- like growth
factor.
• This is done by introducing genes from other species that alter the formation
of this factor and studying the biological effect that result.
2. Study of diseases
• Transgenic animals are designed to increase our understanding of how genes
contribute to the development of diseases.
• These animals are made to serve as models for human diseases so that
investigation of new treatments for diseases is made possible.
• Transgenic models have been developed for many human diseases like
cancer, cystic fibrosis, rheumatoid arthritis and Alzhemer’s disease.
3. Biological products
Transgenic animals that produce useful biological products can be created. This is
done by introduction of the portion of DNA which codes for a particular product.
Examples:-
(i) Human protein (alpha- antitrypsin), used to treat emphysema.
(ii) Human protein enriched milk from transgenic cow, Rosie.
In 1997, the first transgenic cow, Rosie, produced human protein enriched milk.
The milk contained the human alpha- lactalbumin. This milk was nutritionally a
more balanced product for human babies than natural cow milk.
4. Vaccine safety
Transgenic mice are used for the testing of safety of vaccines before they are used on
humans.
Eg:- Polio vaccine is being tested on transgenic mice.
5. Chemical safety testing
• Transgenic animals are made to carry genes which make them more sensitive
to the toxic substances than non- transgenic animals.
• On exposing to the toxic substances, their effects are studied in less time.
BIOETHICS
• It includes a set of standards that may be used to regulate our activities in relation to
the biological world.
• Government of India formed the organization like GEAC ( Genetic Engineering
Approval Committee) to decide the validity and safety of GM organisms.
BIOPIRACY
It is defined as the use of bio resources by multinational companies and other
organizations without proper authorization of the countries and people concerned
without compensatory payment.
Eg:- Basmati rice grown in India.

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