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Application of Recombinant
DNA Technology
Agricultural Biotechnology
An overview
“Our greatest weakness lies in
giving up. The most certain way
to succeed is always to try just
one more time.
- Thomas A. Edison
Content
◍ Agriculture
◍ Agricultural Biotechnology
○ History
◍ Applications
○ Genetic Engeenering
○ Pharmaceutics
○ Nutrition factory
○ Tissue culturing
○ Bioreactors
◍ Benefits
◍ Limitations
◍ Future Aspects
◍ What about Pakistan?
AGRICULTURE
◍ Cultivation of Plants
◍ Agriculture is the
cultivation
of plants and fungi for foo
d, fiber, biofuel, medicinal
plants and other products
used to sustain and
enhance human life.”
• Rich in nutrition
• More Immune to viruses
• Better in quality
• Aid easy and healthy
`cultivation
Agricultural
Biotechnologhy
Agricultural
biotechnology is the term
used in crop and
livestock improvement
through biotechnology
tools. This will focus only
on agricultural crop
biotechnology
Agricultural
biotechnology is the term
used in crop and
livestock improvement
through biotechnology
tools.we will focus only
on agricultural crop
biotechnology
HISTORY
In 1990 -The first
food product of
biotechnology (an
enzyme used in
cheese production
and a yeast used
for baking)
appeared on the
market
In1995, farmers
have been
growing GE crops.
Plant
biotechnology is
founded on the
principles of
cellular totipotency
and genetic
transformation,
which can be
traced back to the
Cell Theory of
Schleiden and
Schwann
and the discovery
of genetic
transformation in
bacteria by
Frederick Griffith,
In 2003,
7 million farmers in
18 countries—more
than 85 percent of
them resource-
poor farmers in the
developing world—
were planting
biotech crops.
Almost one third of
the global biotech
crop area was
grown in
developing
countries..
Applications
Genetic
Engineering
and GM
Crops Pharmaceutics
TISSUE
CULTURING
BIOREACTORS
NUTRITION
FACTORY
Many More
Gene cloning provides a new dimension to crop
breeding by enabling directed changes to be made to
the genotype of a plant, circumventing the random
processes inherent in conventional breeding.
Two general strategies have been used:
Gene addition in which cloning is used to alter the
characteristics of a plant by providing it with one or
more new genes.
Gene subtraction, in which genetic engineering
techniques are used to inactivate one or more of the
plant’s existing genes.
GENE ADDITION
1-Plants that make their own
insecticides
In agricultural settings the greatest problems are
caused by insects.
To reduce losses,crops are regularly sprayed with
insecticides (pyrethroids and
organophosphates)insecticides also have
potentially harmful side effects for members of the
local biosphere chemicals in the ecosystem
sometimes insects escape the toxic effects.
Hight selective and
effective
Biologically degradable
No danger to biospherd
IDEAL INSECTICIDES
BT CROPS
Maize is an example of a crop plant
that is not served well by
conventional insecticides.
A major pest is the European corn
borer (Ostrinia nubilialis), which
tunnels into the plant from eggs laid
on the undersurfaces of leaves,
thereby evading the effects of
insecticides applied by spraying.  Gram +ve bacteria
 cry gene
 protein called delta-endotoxin
 active against a specific
species of target pest.
 highly poisonous to insects,
 80,000 times more toxic than
organophosphate insecticides,
 relatively selective,
 effective against the larvae of
different groups of insects.
Cloning a 4-endotoxin gene in maize
The first attempt at countering this pest by
engineering maize plants to synthesize c-
endotoxin was made by plant
biotechnologists in 1993, working with the
• CryIA(b) version of the toxin.
• Cry protein is 1155 amino acid in length.
• Toxic activity residing in 29-607 amino
acids.
• Rather than isolating the natural gene, a
shortened version containing the first 648
codons was made by artificial gene
synthesis.
• Introduction into the maize embryos.
• Use of tissue culturing
• Introduction of BT crops to Farmers
• Crops were insecticides.
GLYOPHOSATE
In commercial terms the
most important
transgenic plants are
those that have been
engineered to withstand
the herbicide
glyphosate..
Non toxic to insects and
animals
Glyphosate kills all
plants, both weeds and
crop species, and so
has to be applied to
fields very carefully in
order to prevent the
growth of weeds
without harming the
crop itself
PLANT
contain an enzyme
(EPSPS), which converts
shikimate and
phosphoenol pyruvate
(PEP) into
enolpyruvylshikimate-3-
phosphate(EPSP), an
essential precursor for
synthesis of the aromatic
amino acids tryptophan,
tyrosine, and
phenylalanine.
.
Glyphosate competes
with PEP for binding
to the enzyme
surface, thereby
inhibiting synthesis of
EPSP and preventing
the plant from
making the three
amino acids.
As a result
Without these
amino acids, the
plant quickly dies.
.
2-Herbicides and crops
ROUND UP READY CROPS
Monsanto Co. and
called “Roundup
Ready”, reflecting the
trade name of the
herbicide.
Search was carried out for
an organism whose EPSPS
enzyme is resistant to
glyphosate inhibition and
whose EPSPS gene might
therefore be used to confer
resistance on a crop plant.
Place your screenshot here
CROPS
Place your screenshot here
 Glyphosate resistance,
the EPSPS gene from
Agrobacterium strain
CP4 was chosen
 Agrobacterium EPSPS
gene was cloned in a Ti
vector
 Biolistics was used to
introduce the
recombinant vector into
Patunia Alpa callus
culture. After
regeneration, the GM
plants were found to
have a threefold increase
in herbicide resistance.
GENE SUBSTRACTION
mRNA Silencing and the
engineering of fruit
ripening in tomato
Fruit ripening
Effect the colour , taste and texture.
Tissue softening of fruits during ripening is
the result of solubilization of the cell wall by
a group of enzymes.
One of the key enzymes is
polygalacturonase (PG), functioning in
the breakdown of pectin, a polymer of
galacturonic acids that forms part of
the structural support in cell wall.
Flavour
Saver
Antisense RNA and FLAVR SAVR
In normal gene function, the gene is transcribed
into mRNA, which is translated into the enzyme
PG If one can introduce a piece of RNA with a
sequence complementary to that of the PG
mRNA,
RNA would be able to bind to the mRNA
preventing the translation of the mRNA and
consequently the production of the enzyme.
The RNA molecule that is complementary to the
mRNA is called antisense RNA, and the mRNA is
the sense RNA
Nutrition Factory
◍124 million children worldwide are deficient in vitamin A, which leads to
death and blindness
◍b-carotene is precursor to vitamin A & consuming milled rice leads to
vitaminA deficiency.
◍ Mammals make vitamin A from b-carotene, a common carotenoid pigment
normally found in plant photosynthetic membranes.
◍The prototype of golden rice was developed in 2000 and is a light yellow
color
◍ 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’
◍ latest form contains 37 mg/g of carotenoid, of which 84% is b-carotene –
trial.
Golden Rice
◍The Orange cauliflower has
higher than normal levels of b-
carotene that encourages healthy
skin
◍Purple colour comes from
Anthocyanin which may prevent
Heart disease by slowing blood
clotting
◍Tests of the orange cauliflowers
in America found that they
contained 25 times the
concentrations of beta carotene in
normal cauliflowers
Rainbow cauliflower
◍Hirudin is a protein which is
anti coagulant.
◍Found in the sliva of leach
◍Very important
◍Cloning of the hirudin gene to
Brassica napus.
◍Seeds containing Hirudin
◍Isolation of the seeds
◍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 first human clinical trial took
place in 1997. Vaccine against the
toxin from the bacteria E.coli was
produced in potato.
◍Polio vaccine and bannanas
◍Rabies-Tomato plants expressing
rabies antigens could induce
antibodies in mice
◍Cholera-Transgenic potato with
CT-B gene of Vibrio Cholerae
◍Hepatitis B-First human trials of a
potato-based vaccine against
hepatitis B have reported
encouraging results
HIRUDIN
PHARMACEUTICS EDIBLE VACCINES
Micropropagation
Virus free plants
Polen cultures
Somaclonal variations
Protoplast cultures
Conservation of germ lines
Synthetic seeds
TISSUE CULTURING
A device in which a substrate of low value is
utilized by living cells to generate products of
higher value.
BIOREACTORS
 Seed-based plant bioreactors
 Plant Suspension Cultures
 Hairy Root System Bioreactor
• Simple and Cost effective.
• Plant pathogens do not infect humans or animals.
• Easy scale up & rapid harvesting.
• Chimeric plant viruses can be used in production
of vaccines.Produce large biomass.
BENEFITS ◍GM crops are more productive and have a larger yield.
◍Offer more nutritional value and better flavor.
◍ A possibility that they could eliminate allergy-causing properties in some
foods.
◍Inbuilt resistance to pests, weeds and disease. More capable of thriving in
regions with poor soil or adverse climates.
◍ More environment friendly as they require less herbicides and pesticides.
Foods are more resistant and stay ripe for longer so they can be shipped
long distances or kept on shop shelves for longer periods.
◍As more GM crops can be grown on relatively small parcels of land, GM
crops are an answer to feeding growing world populations.Increasing food
production.
◍ improvement and the production of new products with new traits like
◍ improvement in health & safety standard
◍ less environment pollution.
◍ Potential productivity
◍ Medicines and pharmaceutics
◍ Safe
◍ Easy
◍ Liable
LIMITATIONS
Health-related issues:
Allergens and toxins
○People with food allergies usually react only to one or a few
allergens in one or two specific foods.
○A major safety concern raised with regard to genetic engineering
technology is the risk of introducing allergens and toxins into
otherwise safe foods.
Social issues
“Terminator” technology Most farmers buy fresh seeds each
season, particularly of such crops as corn, green peppers, and
tomatoes.
Anyone growing hybrid varieties must buy new seeds annually,
because seeds from last year’s hybrids grown on the farm will
not produce plants identical to the parent.
There is a belief among some opponents of genetic engineering
technology that transgenic crops might cross pollinate with
related weeds, possibly resulting in “superweeds” that become
more difficult to control.
Environmental issues:
Future Aspects
◍Genetically-modifiedfoods have the potential to solve many of the
world'shunger and malnutritionproblems, and to help protect and
preserve the environment by increasing yield and reducing reliance
upon chemical pesticides and herbicides.
◍ Yet there are many challenges ahead for governments, especially in
the areas of safety testing, regulation, international policy and food
labeling.
◍Many people feel that genetic engineering is the inevitable wave of
the future and that we cannot afford to ignore a technology that has
such enormous potential benefits.
◍However, we must proceed with caution to avoid causing
unintended harm to human health and the environment as a result of
our enthusiasm for this powerful technology.
◍ At the present time,genetically modified foodsare dangerous for the
world that we live in, not just for its people, but the delicate balance
between organisms that inhabit it..
Pakistan and Biotechnology
Pakistan is an agricultural country yet the
applications of biotechnology are less then other
countries.
In 2005 BT cotton was introduced but farmers
couldn’t get enough benefit , it was the terminator
technology which became the reason of loss, that’s
why we need to provide ethical education to the
farmers.
Moreover GM crops can play a better role..
Through GM crops we can avoid Food Adultration
that has more bad effects on our health.
Pakistan has more threats from food Adultratiion.
For now as an indicidual we can start Kitchen
gardening
Organic food is more important.

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Agricultural biotechnology, overview

  • 1. Application of Recombinant DNA Technology Agricultural Biotechnology An overview
  • 2. “Our greatest weakness lies in giving up. The most certain way to succeed is always to try just one more time. - Thomas A. Edison
  • 3. Content ◍ Agriculture ◍ Agricultural Biotechnology ○ History ◍ Applications ○ Genetic Engeenering ○ Pharmaceutics ○ Nutrition factory ○ Tissue culturing ○ Bioreactors ◍ Benefits ◍ Limitations ◍ Future Aspects ◍ What about Pakistan?
  • 4. AGRICULTURE ◍ Cultivation of Plants ◍ Agriculture is the cultivation of plants and fungi for foo d, fiber, biofuel, medicinal plants and other products used to sustain and enhance human life.” • Rich in nutrition • More Immune to viruses • Better in quality • Aid easy and healthy `cultivation
  • 5. Agricultural Biotechnologhy Agricultural biotechnology is the term used in crop and livestock improvement through biotechnology tools. This will focus only on agricultural crop biotechnology Agricultural biotechnology is the term used in crop and livestock improvement through biotechnology tools.we will focus only on agricultural crop biotechnology
  • 6. HISTORY In 1990 -The first food product of biotechnology (an enzyme used in cheese production and a yeast used for baking) appeared on the market In1995, farmers have been growing GE crops. Plant biotechnology is founded on the principles of cellular totipotency and genetic transformation, which can be traced back to the Cell Theory of Schleiden and Schwann and the discovery of genetic transformation in bacteria by Frederick Griffith, In 2003, 7 million farmers in 18 countries—more than 85 percent of them resource- poor farmers in the developing world— were planting biotech crops. Almost one third of the global biotech crop area was grown in developing countries..
  • 8. Gene cloning provides a new dimension to crop breeding by enabling directed changes to be made to the genotype of a plant, circumventing the random processes inherent in conventional breeding. Two general strategies have been used: Gene addition in which cloning is used to alter the characteristics of a plant by providing it with one or more new genes. Gene subtraction, in which genetic engineering techniques are used to inactivate one or more of the plant’s existing genes.
  • 9. GENE ADDITION 1-Plants that make their own insecticides In agricultural settings the greatest problems are caused by insects. To reduce losses,crops are regularly sprayed with insecticides (pyrethroids and organophosphates)insecticides also have potentially harmful side effects for members of the local biosphere chemicals in the ecosystem sometimes insects escape the toxic effects. Hight selective and effective Biologically degradable No danger to biospherd IDEAL INSECTICIDES
  • 10. BT CROPS Maize is an example of a crop plant that is not served well by conventional insecticides. A major pest is the European corn borer (Ostrinia nubilialis), which tunnels into the plant from eggs laid on the undersurfaces of leaves, thereby evading the effects of insecticides applied by spraying.  Gram +ve bacteria  cry gene  protein called delta-endotoxin  active against a specific species of target pest.  highly poisonous to insects,  80,000 times more toxic than organophosphate insecticides,  relatively selective,  effective against the larvae of different groups of insects.
  • 11. Cloning a 4-endotoxin gene in maize The first attempt at countering this pest by engineering maize plants to synthesize c- endotoxin was made by plant biotechnologists in 1993, working with the • CryIA(b) version of the toxin. • Cry protein is 1155 amino acid in length. • Toxic activity residing in 29-607 amino acids. • Rather than isolating the natural gene, a shortened version containing the first 648 codons was made by artificial gene synthesis. • Introduction into the maize embryos. • Use of tissue culturing • Introduction of BT crops to Farmers • Crops were insecticides.
  • 12. GLYOPHOSATE In commercial terms the most important transgenic plants are those that have been engineered to withstand the herbicide glyphosate.. Non toxic to insects and animals Glyphosate kills all plants, both weeds and crop species, and so has to be applied to fields very carefully in order to prevent the growth of weeds without harming the crop itself PLANT contain an enzyme (EPSPS), which converts shikimate and phosphoenol pyruvate (PEP) into enolpyruvylshikimate-3- phosphate(EPSP), an essential precursor for synthesis of the aromatic amino acids tryptophan, tyrosine, and phenylalanine. . Glyphosate competes with PEP for binding to the enzyme surface, thereby inhibiting synthesis of EPSP and preventing the plant from making the three amino acids. As a result Without these amino acids, the plant quickly dies. . 2-Herbicides and crops
  • 13. ROUND UP READY CROPS Monsanto Co. and called “Roundup Ready”, reflecting the trade name of the herbicide. Search was carried out for an organism whose EPSPS enzyme is resistant to glyphosate inhibition and whose EPSPS gene might therefore be used to confer resistance on a crop plant. Place your screenshot here CROPS
  • 14. Place your screenshot here  Glyphosate resistance, the EPSPS gene from Agrobacterium strain CP4 was chosen  Agrobacterium EPSPS gene was cloned in a Ti vector  Biolistics was used to introduce the recombinant vector into Patunia Alpa callus culture. After regeneration, the GM plants were found to have a threefold increase in herbicide resistance.
  • 15. GENE SUBSTRACTION mRNA Silencing and the engineering of fruit ripening in tomato Fruit ripening Effect the colour , taste and texture. Tissue softening of fruits during ripening is the result of solubilization of the cell wall by a group of enzymes. One of the key enzymes is polygalacturonase (PG), functioning in the breakdown of pectin, a polymer of galacturonic acids that forms part of the structural support in cell wall. Flavour Saver
  • 16. Antisense RNA and FLAVR SAVR In normal gene function, the gene is transcribed into mRNA, which is translated into the enzyme PG If one can introduce a piece of RNA with a sequence complementary to that of the PG mRNA, RNA would be able to bind to the mRNA preventing the translation of the mRNA and consequently the production of the enzyme. The RNA molecule that is complementary to the mRNA is called antisense RNA, and the mRNA is the sense RNA
  • 17.
  • 18. Nutrition Factory ◍124 million children worldwide are deficient in vitamin A, which leads to death and blindness ◍b-carotene is precursor to vitamin A & consuming milled rice leads to vitaminA deficiency. ◍ Mammals make vitamin A from b-carotene, a common carotenoid pigment normally found in plant photosynthetic membranes. ◍The prototype of golden rice was developed in 2000 and is a light yellow color ◍ 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’ ◍ latest form contains 37 mg/g of carotenoid, of which 84% is b-carotene – trial. Golden Rice
  • 19. ◍The Orange cauliflower has higher than normal levels of b- carotene that encourages healthy skin ◍Purple colour comes from Anthocyanin which may prevent Heart disease by slowing blood clotting ◍Tests of the orange cauliflowers in America found that they contained 25 times the concentrations of beta carotene in normal cauliflowers Rainbow cauliflower
  • 20. ◍Hirudin is a protein which is anti coagulant. ◍Found in the sliva of leach ◍Very important ◍Cloning of the hirudin gene to Brassica napus. ◍Seeds containing Hirudin ◍Isolation of the seeds ◍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 first human clinical trial took place in 1997. Vaccine against the toxin from the bacteria E.coli was produced in potato. ◍Polio vaccine and bannanas ◍Rabies-Tomato plants expressing rabies antigens could induce antibodies in mice ◍Cholera-Transgenic potato with CT-B gene of Vibrio Cholerae ◍Hepatitis B-First human trials of a potato-based vaccine against hepatitis B have reported encouraging results HIRUDIN PHARMACEUTICS EDIBLE VACCINES
  • 21. Micropropagation Virus free plants Polen cultures Somaclonal variations Protoplast cultures Conservation of germ lines Synthetic seeds TISSUE CULTURING
  • 22. A device in which a substrate of low value is utilized by living cells to generate products of higher value. BIOREACTORS  Seed-based plant bioreactors  Plant Suspension Cultures  Hairy Root System Bioreactor • Simple and Cost effective. • Plant pathogens do not infect humans or animals. • Easy scale up & rapid harvesting. • Chimeric plant viruses can be used in production of vaccines.Produce large biomass.
  • 23. BENEFITS ◍GM crops are more productive and have a larger yield. ◍Offer more nutritional value and better flavor. ◍ A possibility that they could eliminate allergy-causing properties in some foods. ◍Inbuilt resistance to pests, weeds and disease. More capable of thriving in regions with poor soil or adverse climates. ◍ More environment friendly as they require less herbicides and pesticides. Foods are more resistant and stay ripe for longer so they can be shipped long distances or kept on shop shelves for longer periods. ◍As more GM crops can be grown on relatively small parcels of land, GM crops are an answer to feeding growing world populations.Increasing food production. ◍ improvement and the production of new products with new traits like ◍ improvement in health & safety standard ◍ less environment pollution. ◍ Potential productivity ◍ Medicines and pharmaceutics ◍ Safe ◍ Easy ◍ Liable
  • 24.
  • 25. LIMITATIONS Health-related issues: Allergens and toxins ○People with food allergies usually react only to one or a few allergens in one or two specific foods. ○A major safety concern raised with regard to genetic engineering technology is the risk of introducing allergens and toxins into otherwise safe foods. Social issues “Terminator” technology Most farmers buy fresh seeds each season, particularly of such crops as corn, green peppers, and tomatoes. Anyone growing hybrid varieties must buy new seeds annually, because seeds from last year’s hybrids grown on the farm will not produce plants identical to the parent. There is a belief among some opponents of genetic engineering technology that transgenic crops might cross pollinate with related weeds, possibly resulting in “superweeds” that become more difficult to control. Environmental issues:
  • 26. Future Aspects ◍Genetically-modifiedfoods have the potential to solve many of the world'shunger and malnutritionproblems, and to help protect and preserve the environment by increasing yield and reducing reliance upon chemical pesticides and herbicides. ◍ Yet there are many challenges ahead for governments, especially in the areas of safety testing, regulation, international policy and food labeling. ◍Many people feel that genetic engineering is the inevitable wave of the future and that we cannot afford to ignore a technology that has such enormous potential benefits. ◍However, we must proceed with caution to avoid causing unintended harm to human health and the environment as a result of our enthusiasm for this powerful technology. ◍ At the present time,genetically modified foodsare dangerous for the world that we live in, not just for its people, but the delicate balance between organisms that inhabit it..
  • 27. Pakistan and Biotechnology Pakistan is an agricultural country yet the applications of biotechnology are less then other countries. In 2005 BT cotton was introduced but farmers couldn’t get enough benefit , it was the terminator technology which became the reason of loss, that’s why we need to provide ethical education to the farmers. Moreover GM crops can play a better role.. Through GM crops we can avoid Food Adultration that has more bad effects on our health. Pakistan has more threats from food Adultratiion. For now as an indicidual we can start Kitchen gardening Organic food is more important.

Editor's Notes

  1. Later on Herbicide soybean,cotton,corn and many more crops were developed.