INSTA LEARN
PLUS TWO
BOTANY
BIOTECHNOLOGY
AND ITS APPLICATIONS
Totipotency
BIOTECHNOLOGY PRINCIPLES
Capacity to generate a whole plant from any
cell/explant is called totipotency.
Green revolution
The Green Revolution succeeded in tripling
the food supply but yet it was not enough
to feed the growing human population. In-
creased yields have partly been due to the
use of improved crop varieties, but mainly
due to the use of better management prac-
tices and use of agrochemicals (fertilisers
and pesticides).
Micro propagation
By application of tissue culture methods it is
possible to achieve propagation of a large
number of plants in very short durations. This
method of producing thousands of plants
through tissue culture is called micro-prop-
agation.
Tissue Culture
A method of biological research in which
fragments a plant are transferred to an artifi-
cial environment can continue to survive and
function.
Somaclones
Each plants produced through tissue culture
will be genetically identical to the original
plant from which they were grown, i.e., they
are somaclones
Explants
Any part of a plant taken out and grown in a
test tube, under sterile conditions in special
nutrient media.
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Meristem culture What is GMO?
Even if the plant is infected with a virus, the Plants, bacteria, fungi and animals whose
meristem (apical and axillary) is free of virus. genes have been altered by manipulation
Hence, one can remove the meristem and are called Genetically Modified Organisms
grow it in vitro to obtain virus-free plants. (GMO).
GM plants have been useful in many ways.
Somatic hybridisation Genetic modification has:
(i) made crops more tolerant to abiotic
Isolated protoplasts from two different va- stresses (cold, drought, salt, heat).
rieties of plants - each having a desirable
character - can be fused to get hybrid proto- (ii) reduced reliance on chemical pesticides
plasts, which can be further grown to form a (pest-resistant crops).
new plant. These hybrids are called somatic (iii) helped to reduce post harvest losses.
hybrids while the process is called somatic
(iv) increased efficiency of mineral usage by
hybridisation.
plants (this prevents early exhaustion of
fertility of soil).
(v) enhanced nutritional value of food, e.g.,
golden rice, i.e., Vitamin ‘ A ‘ enriched rice.
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Pest Resistant Plants - Bt Cotton ⇒ Some
strains of Bacillus thuringiensis have proteins
that kill insects like lepidopterans (tobac-
co budworm, armyworm). ⇒ B. thuringien-
sis forms a toxic insecticidal protein (Bt tox-
in) crystal during a particular phase of their
growth. It does not kill the Bacillus as it exists
as inactive protoxins.
Nematode resistance in tobacco
plants
A nematode Meloidegyne incognita infects
the roots of tobacco plants and causes a
great reduction in yield. RNA interference
(RNAi) strategy is used to prevent this infes-
tation.
RNAi is a method of cellular defense in all eu-
karyotic organisms. It prevents translation of
a specific mRNA (silencing) due to a comple-
mentary dsRNA molecule.
When an insect ingest the inactive toxin, it is
converted into active toxin due to the alkaline
pH of the gut which solubilise the crystals. The
toxin binds to the surface of midgut epithelial
cells and creates pores. It causes cell swell-
ing and lysis and death of the insect.
Bt toxin genes were isolated from B. thuring-
iensis and incorporated into crop plants such
as cotton.
⇒ Most Bt toxins are insect-group specific.
The toxin is coded by a gene named cry . E.g.
the proteins encoded by the genes crylAc
and cryllAb control the cotton bollworms that
of crylAb controls corn borer
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⇒ Source from an infection by RNA viruses
or mobile genetic elements (transposons)
that replicate via an RNA intermediate. Using
Agrobacterium vectors, nematode-specific
genes (DNA) were introduced into the host
plant. It produced both sense and anti sense
RNA as these RNA are complementary to
each other, they bind to form double strand-
ed RNA that initiate RNAi.
Genetically Engineered Insulin
It is possible to produce human insulin using
bacteria Insulin consists of two short polypep- Gene Therapy
tide chains (chain A & chain B) that are linked
together by disulphide bridges. In mammals, Method to correct a gene defect diagnosed
insulin is synthesized as a pro-hormone. The in a child/embryo
pro-hormone needs processing before it
becomes a fully mature and functional hor- First Clinical gene therapy was given to a
mone. The prohormone contains an extra 4-year old girl with adenosine deaminase
stretch called the C peptide. This is removed (ADA) deficiency. The disorder is caused due
during maturation into insulin, to the deletion of the gene for adenosine de-
aminase.
This can be cured by bone marrow trans-
plantation or by enzyme replacement ther-
apy (injection of functional ADA). But these
approaches are not completely curative.
In gene therapy, lymphocytes from the pa-
tient’s blood are grown in a culture. Then, a
functional ADA cDNA (using a retroviral vec-
tor) is introduced into these lymphocytes.
Then, they are returned to the patient. How-
ever, if the ADA gene (from marrow cells)
is introduced into cells at early embryonic
stages, it could be a permanent cure
MOLECULAR DIAGNOSIS
In 1983, Eli Lilly an American company pre- Recombinant DNA technology, PCR and En-
pared two DNA sequences corresponding to zyme Linked Immunosorbent Assay (ELISA)
A & B chains of human insulin and introduced are some techniques for early diagnosis
them in plasmids of E.coli to produce insulin However, very low concentration of a bacte-
chains. Chains A & B were produced sepa- ria or virus can be detected by amplification
rately, extracted and combined by creating of their nucleic acid by PCR PCR is used to
disulfide bonds to form human insulin. detect HIV in suspected AIDS patients, muta-
tions in genes in suspected cancer patients.
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TRANSGENIC ANIMALS PROBLEMS OF PATENT:
E.g. Basmati rice, herbal medicines like tur-
These are the animals whose genome has meric, neem etc. India has 27 varieties of
been altered by introduction of an extra (for- Basmati. In 1997, an American company got
eign) gene by manipulation E.g. Transgenic patent rights on Basmati rice through the US
rats, rabbits, pigs, sheep cows and fish. Patent and Trademark Office. This allowed
the company to sell a ‘new’ variety of Bas-
BIOLOGICAL PRODUCTS:
mati. This had actually been derived from
Human protein ( α-l-antitrypsin) used to Indian farmer’s varieties. Indian Basmati
treat emphysema, products for treatment of was crossed with semi-dwarf varieties and
phenylketonuria claimed as a novelty. Other people selling
Basmati rice could be restricted by the pat-
(PKU) and cystic fibrosis etc. in 1997, Rosie ent
(first transgenic cow)
produced human protein-enriched milk (2.4
gram per litre).
ETHICAL ISSUES
Problem Indian Government has set up or-
ganizations like GEAC (Genetic Engineering
Approval committee), which make decisions
about the validity of GM research and the
safety of GM-organisms for public services.
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Biopiracy is the term used to refer to the use
of bio-resources by multinational compa-
nies and other organisations without proper Which of the following is a curative method of
authorisation from the countries and people ADA deficiency?
concerned without compensatory payment.
(a) ELISA
(b) Autoradiography
(c) PCR
(d) Bonemarrowtransplantation
The Indian Parliament has recently cleared
the second amendment of the Indian Patents
Bill, that takes such issues into consideration,
including patent terms emergency provisions
and research and development initiative.
Name the ‘cry’ gene that control corn borer in-
sects
Space for Keynotes
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Fill in the blank: Explain the terms:
Animals that have had their DNA manipulat- (a) Micropropagation (b) Totipotency
ed to possess and express an extra gene are
known as .........
Observe the diagram and answer A and B . Bt cotton is a genetically modified plant.
(a) What does ‘Bt’ stand for?
(b) What is the specific feature of ‘Bt’ cotton
plant?
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Match the following: (a) Define transgenic animals.
(b) Write any two uses of transgenic animals
A B
Bt Cotton Antigen-antibody
reaction
Meloidegyne incog- Cry gene
nitia
Gene Therapy RNA interference
ELISA ADA deficiency
Explain how Eli Lilly company produced genet- Gene therapy is a corrective therapy for a her-
ically engineered insulin iditary disease.
(a) Name the disease which was successful-
ly corrected by gene therapy for the first
time.
(b) How gene therapy is practiced for a per-
manent cure of the disease ?
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Match the Column A with Column B:
Column A Column B
(a) Human Alpha (1) ELISA
lactalbumin
(b) Antigen Antibody (2) ELI LILY
interaction
(c) Genetically engi- (3) CORN BORER
neered Insulin
(d) Cry IAb (4) ROSIE
(5) BOLL WORM
Space for Keynotes
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