Success Story of Transgenic crops Important Traits for Crop Improvement High crop yield  High nutritional quality  Abiotic stress tolerance  Pest resistance  Adaptation to.

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Transcript Success Story of Transgenic crops Important Traits for Crop Improvement High crop yield  High nutritional quality  Abiotic stress tolerance  Pest resistance  Adaptation to.

Success Story of
Transgenic crops
Important Traits for Crop
Improvement
High crop yield
 High nutritional quality
 Abiotic stress tolerance
 Pest resistance
 Adaptation to inter-cropping
 Nitrogen Fixation
 Insensitivity to photo-period
 Elimination of toxic compounds

HYBRID DEVELOPMENT
FOR HIGHER YIELD
NUTRITIONAL
QUALITY
BIOTIC STRESS
TOLERANCE
TRANSGENIC PLANTS
ENHANCED
SHELF LIFE
ABIOTIC STRESS
TOLERANCE
INDUSTRIAL
PRODUCTS
PHARMACEUTICALS
& EDIBLE VACCINE
1985
1988
1992
1st transgenic plants produced
Particle bombardment developed
1994
Flavr-Savr tomato is released
1996
Herbicide- and insect-resistant crops approved for cultivation
GM crops considered substantially equivalent to hybrid varieties
4.3 million acres of GM crops planted
1998
GM food is dangerous (UK TV)
Monarch butterfly paper causes uproar
1999
GM corn is excluded from its baby food
Greenpeace starts anti-GM campaign
75 million acres of GM crops planted
2000
Golden rice with ß-carotene developed
McDonald’s rejects GM potatoes
Tearless Onion
Colorful Cauliflowers
Purple tomatoes
Blue Roses
The big five successful traits
 Insect Resistance
 Delayed Fruit Ripening
 Nutritional Enhancing
 Herbicide Resistance
 Virus Resistance
Herbicide Resistance
 Glyphosate Resistance
i. Glyphosate = “Roundup”, “Tumbleweed” = Systemic herbicide
ii. Marketed under the name Roundup, glyphosate inhibits the enzyme
EPSPS (S-enolpyruvlshikimate-3 phosphate – involved in chloroplast
amino acid synthesis), makes aromatic amino acids.
iii. The gene encoding EPSPS has been transferred from glyphosateresistant E. coli into plants, allowing plants to be resistant.
 Glufosinate Resistance
i.
Glufosinate (the active ingredient being phosphinothricin) mimics the
structure of the amino acid glutamine, which blocks the enzyme
glutamate synthase.
ii. Plants receive a gene from the bacterium Streptomyces (bar gene) that
produce a protein that inactivates the herbicide.
Herbicide Resistance
 Bromoxynil Resistance
i. A gene encoding the enzyme bromoxynil nitrilase (BXN) is
transferred from Klebsiella pneumoniae bacteria to plants.
ii. Nitrilase inactivates the Bromoxynil before it kills the plant.
 Sulfonylurea.
i. Kills plants by blocking an enzyme needed for synthesis of
the amino acids valine, leucine, and isoleucine.
ii. Resistance generated by mutating a gene in tobacco plants
(acetolactate synthase), and transferring the mutated gene
into crop plants.
Roundup Ready™ Soybeans
A problem in agriculture is the reduced growth of crops imposed by
the presence of unwanted weeds. Herbicides such as RoundupTM and
Liberty LinkTM are able to kill a wide range of weeds and have the
advantage of breaking down easily. Development of herbicide
resistant crops allows the elimination of surrounding weeds without
harm to the crops.
ROUNDUP (Glyphosate) RESISTANCE
( HERBICIDE TOLERANCE IN CROPS)
Glucose
ROUNDUP
(Glyphosate )
3 phosphoglycerate
Tryptophan
Glycolysis
EPSP SYNTHASE
Phosphoenol
pyruvate
EPSP SYNTHASE
Tyrosine
Phenylalaline
ROUNDUP
(Glyphosate )
In transgenic plant, herbicide cannot bind the mutant of EPSPS
(Roundup resistant cotton and soybean)
Insect resistance
Anti-Insect Strategy - Insecticides
a) Toxic crystal protein from Bacillus thuringensis



Toxic crystals found during sporulation
Alkaline protein degrades gut wall of lepidopteran larvae
• Corn borer catepillars
• Cotton bollworm catepillars
• Tobacco hornworm catepillars
• Gypsy moth larvae
Sprayed onto plants – but will wash off
The Bt toxin isolated from Bacillus thuringiensis has been used in plants.
The gene has been placed in corn, cotton, and potato, and has been
marketed.
Insect Resistance
Various insect resistant crops have been produced. Most of
these make use of the Cry gene in the bacteria Bacillus
thuringiensis (Bt); this gene directs the production of a protein
that causes paralysis and death to many insects.
Corn hybrid with a Bt gene
Corn hybrid susceptible to European
corn borer
δ -endotoxin gene (Cry gene) of Bacillus thuriengenesis
GENE FOR Bt TOXIN WAS TRANSFERRED
TO OBTAIN BT TRANSGENIC PLANTS
PLANT SYNTHESIZES INACTIVE PROTOXIN
INSECT FEEDS ON
TRANSGENIC PLANT
PROTEINASE
DIGESTION IN
INSECT GUT
MAKES THE
ACTIVE TOXIN
Toxin binds a receptor on the gut epithelial cells, forms a channel
on the membrane. This causes electrolyte leakage and insect death
Virus resistance
a) Plants may be engineered with genes for resistance to viruses,
bacteria, and fungi.
b) Virus-resistant plants have a viral protein coat gene that is
overproduced, preventing the virus from reproducing in the host
cell, because the plant shuts off the virus’ protein coat gene in
response to the overproduction.
c) Coat protein genes are involved in resistance to diseases such as
cucumber mosaic virus, tobacco rattle virus, and potato virus X.
Delayed Fruit Ripening
a) Allow for crops, such as tomatoes, to have a higher shelf
life.
b) Tomatoes generally ripen and become soft during
shipment to a store.
c) Tomatoes are usually picked and sprayed with the plant
hormone ethylene to induce ripening, although this does
not improve taste
d) Tomatoes have been engineered to produce less
ethylene so they can develop more taste before ripening,
and shipment to markets
Delayed Fruit Ripening
What happened to the Flavr Savr tomato?
i. Produced by blocking the polygalacturonase (PG) gene, which is
involved in spoilage. PG is an enzyme that breaks down pectin, which
is found in plant cell walls.
ii. Plants were transformed with the anti-sense PG gene, which is mRNA
that base pair with mRNA that the plant produces, essentially blocking
the gene from translation.
iii.First genetically modified organism to be approved by the FDA, in
1994.
iv.Tomatoes were delicate, did not grow well in Florida, and cost much
more than regular tomatoes.
v. Calgene was sold to Monsanto after Monsanto filed a patentinfringement lawsuit against Calgene, and the Flavr Savr tomato left the
market.
First biotech plant product – Flav’r Sav’r
tomato
“Rot-Resistant Tomato”
Anti-sense gene  complementary to polygalacturonase (PG)
PG = pectinase  accelerates plant decay/rotting
Nutritionally Enhanced
Plants
1.More than one third of the world’s population relies on
rice as a food staple, so rice is an attractive target for
enhancement.
2.Golden Rice was genetically engineered to produce high
levels of beta-carotene, which is a precursor to vitamin
A. Vitamin A is needed for proper eyesight.
3.Other enhanced crops include iron-enriched rice and
tomatoes with three times the normal amount of betacarotene
Golden Rice
Normal rice
Transgenic technology produced a type of rice that
accumulates beta-carotene in rice grains. Once inside the body,
beta-carotene is converted to vitamin A.
“Normal” rice
“Golden” rice
Gernayl Gernayl diphosphate (GGPP)
Phytoene synthase
Phytoene
Phytoene desaturase
Lycopene
Lycopene cyclase
Beta carotene
Complete biochemical pathway in the rice for production of
beta-carotene, a precursor for vitamin A
The prototype of golden rice was developed in 2000 and is a
light yellow color (b). It contains 1.6 mg/g of carotenoid.
In 2005, new transgenic lines were developed that dramatically
increased the amount of carotenoid synthesized, making the
rice a deep golden color (c).
This latest form contains 37 mg/g of carotenoid, of which
84% is b-carotene – trial
Molecular Farming
1.A new field where plants and animals are genetically engineered to
produce important pharmaceuticals, vaccines, and other valuable
compounds.
2.Plants may possibly be used as bioreactors to mass-produce
chemicals that can accumulate within the cells until they are
harvested.
3.Soybeans have been used to produce monoclonal antibodies with
therapeutic value for the treatment of colon cancer. Drugs can
also be produced in rice, corn, and tobacco plants
4.Plants have been engineered to produce human antibodies against
HIV. Pharmaceuticals has begun clinical trials with herpes
antibodies produced in plants.
Why Plants?
Plants offer unique benefits for the production of
pharmaceutical proteins:

Ease of Scale-up



Low Cost
Reduced Capital Expenditures
No Animal Contaminants (virus)
Molecular Farming
(Vaccines)
a) Making plants that produce vaccines
b) Potatoes have been studied using a portion of the E.
coli enterotoxin in mice and humans.
c) Other candidates for edible vaccines include banana
and tomato, and alfalfa, corn, and wheat are possible
candidates for use in livestock.
d) Edible vaccines may lead to the eradication of
diseases such as hepatitis B and polio.
Pharmaceutical Production in Plants
Genetically modified plants have been used as “bioreactors” to produce
therapeutic proteins for more than a decade. A recent contribution by
transgenic plants is the generation of edible vaccines.
Edible vaccines are vaccines produced in plants that can be administered
directly through the ingestion of plant materials containing the vaccine. Eating
the plant would then confer immunity against diseases.
Edible vaccines produced by transgenic plants are
attractive for many reasons. The cost associated
with the production of the vaccine is low, especially
since the vaccine can be ingested directly, and
vaccine production can be rapidly up scaled should
the need arises. Edible vaccine is likely to reach
more individuals in developing countries.
The first human clinical trial took place in 1997.
Vaccine against the toxin from the bacteria E.coli
was produced in potato. Ingestion of this transgenic
potato resulted in satisfactory vaccinations and no
adverse effects.
Edible Vaccines
One focus of current vaccine effort is on hepatitis B, a virus responsible for causing
chromic liver disease. Transgenic tobacco and potatoes were engineered to express
hepatitis B virus vaccine. During the past two years, vaccines against a E.coli toxin, the
respiratory syncytial virus, measles virus, and the Norwalk virus have been
successfully expressed in plants and delivered orally. These studies have supported the
potential of edible vaccines as preventive agents of many diseases.
There is hope to produce edible vaccines in bananas, which are grown extensively
throughout the developing world.
Molecular Farming
(Biopolymers and Plants)
a) Plant seeds may be a potential source for plastics that
could be produced and easily extracted.
b) A type of PHA (polyhydroxylalkanoate) polymer called
“poly-beta-hydroxybutyrate”, or PHB, is produced in
Arabidopsis, or mustard plant.
c) PHB can be made in canola seeds by the transfer of three
genes from the bacterium Alicaligenes eutrophus, which
codes for enzymes in the PHB synthesis pathway.
d) A polymer called PHBV produced through Alicaligenes
fermentation, which is sold under the name Biopol.