The document discusses the concepts of transgenic plants and biotechnology, highlighting the creation and benefits of genetically modified organisms (GMOs) through various gene transfer methods. It provides a historical timeline of significant advancements in plant genetic engineering and outlines the advantages and disadvantages of transgenic crops and GM foods. Additionally, it explores biopharming, the use of plants for pharmaceutical production, and addresses the risks and regulatory concerns associated with genetic modification.

1. Plants Biotechnology

2. TRANSGENIC PLANTS:
Terms To Know
TRANSGENE:
It is foreign gene or genetic material that has been transferred naturally or by any of number of
genetic engineering techniques from one organism to another.
TRANSGENESIS:
The phenomenon of introduction of exogenous DNA into the genome to create and maintain a stable
and heritable character.
TRANSGENIC PLANTS:
The plant whose genome is altered by adding one or more transgenesare known as transgenic plants.

3. HISTORY:
• 1st transgenic plant produced which is an antibiotic
resistance tobacco plant.
1982
• 1st successful plant genetic engineering experiments
using caulimovirus vector.
1984
• 1st genetically modified crop approved for sale in U.S
was FlavrSavr tomato.
1994
• 1st pesticide producing crop,Bt Potato was approved by
U.S Environmental Protection Agency .
1995
• 1st genetically modified flower Moondust ,bluish
colored camation ,was introduced .
1996
• Golden rice with beta-carotene developed with
increased nutrient value.
2000

4. SOME EXAMPLES OF TRANSGENIC
PLANTS

5. WHY DO WE NEED TRANSGENIC
PLANTS?
• Improvement of agriculture value of plant (resistance to insect attack)
• Living bioreactor ->Produce specific protein
• Studying action of genes during development or other biological
process

6. GENE TRANSFER METHODS
BIOLOGICAL METHODS:
 Agrobacterium medicated gene transfer
 Plants virus vectors
PHYSICAL METHODS:
 Microinjection
 Micro projectile
 Liposome fusion
 Electroporation
Chemical Methods:
 Polyethylene glycol mediated
 Diethylaminoethyl dextran mediated

9. Advantages And Disadvantages Of
Transgenic Plants
Advantages
• Plants cells are totipotent-
>whole plant can be regenerated
from a single cell
• Plants have many off springs -
>rare combinations and mutation
can be founded
Disadvantages
• Plant regenerating from single
cells or not genetically
homogenous(genetically
instable)
• Large genomes (polypoid-
>presence of many genomes in
one cell)

10. Transgenic Crops:
• A transgenic crop is a genetically modified organism (GMO).
• Transgenic indicates that a transfer of genes has occurred using
recombinant DNA technology.
• Generally a transgenic crop contains one or more genes that have
been inserted artificially either from an unrelated plant or from
different species altogether.

11. Gene Guns:
• Gene guns (also known as biolistics) "shoot” target
genes into plant cells.
• DNA is bound to tiny particles of gold or tungsten
which are subsequently shot into plant tissue or single
plant cells under high pressure. The accelerated
particles penetrate both the cell wall and membranes.
• The DNA separates from the metal and is integrated
into plant DNA inside the nucleus. This method has
been applied successfully for many cultivated crops,
especially monocots like wheat or maize.
• The major disadvantage of this procedure is that
serious damage can be done to the cellular tissue.

12. Process of GeneGun:

13. Advantages of Transgenic Crops:
Transgenic crops are developed according to needs of human beings and to
protect from the harmful organisms. So, majorly it has benefits like:
• Transgenic crops are resistant to various diseases and reduce the investment
cost on chemicals and more labor force.
• High product yielding in lesser time and also reduces the food scarcity.
• Advanced farming techniques are developed.
• Some African countries are facing nutritious food scarcity, to avoid such
problems crops are modified to more nutrients by genetic transformation.
• These transgenic crops can be stored for long time due to its increased
resistance to spoilage.
• These crops can also grow under drought conditions.

14. What are GM Foods?
• GM Foods (genetically modified foods) also known as
genetically engineered foods or bioengineered foods.
• Produce from such organisms whom DNA is altered
through genetic engineering.
• Involves the insertion of DNA from one organism into
another.

15. History
• In 1946 scientist discover that DNA can transfer
between organisms.
• In 1994 first GM plant approved for food use was
Flavr Savr tomato.
• In 2015 first GM animal approved for food use was
Aqu Advantage Salmon

16. Process:
• At first scientist need an organism which contain the gene of interest.
• Then they extract the DNA from that organism.
• Then with the help of gene cloning, they isolate the gene of interest
from DNA.
• Then they modify that gene in the labs.
• Newly modified transgene is ready to insert in the targeted organism.
• This is done by two methods.

17. • Agrobacterium:
A type of bacterium which can naturally transfer
transgenes
to the nucleus of targeted organism.
• Genegun:
It shoots the golden particle coated with copies of transgene .
If the transgene is successfully integrated then a new organism with such
trait will grown.
• Here the genetic engineering is done .
• These transgenic organisms are handed over the breeders .So they breed it
with other such organisms in more traditional ways, to select for other
desirable traits.

19. Examples:
• Tomatoes
• Honey
• Soybean
• Canola
• Potatoes
• Papaya

20. Advantages and disadvantages of GM foods:
Advantages
• GM crops use less water.
• They take less land to grow more
food.
• They can save core crops from
extinction.
• They can conserve energy ,soil
and water resources.
Disadvantages
• There may be an increased risk
of allergies and
food intolerance.
• Animal protein could be affected
by them.
• They encourages use of
additional herbicides.
• They can contaminate other
fields.

21. BIOPHARMING

22. • What is biopharming?
• Why use plants?
• Current and evolving regulation
• What are the risks and concerns?

23. What is biopharming?
• The use of agricultural plants for the production of useful molecules
for non food, feed or fiber applications. (also called molecular
farming, pharming, or biopharming)
• Plants are already grown to produce valuable molecules, including
many drugs.
• Biopharming is different because the plants are genetically engineered
(GE) to produce the molecules we want them to.

24. Plant Products:
1. Plant derived pharmaceuticals (non-GE)
• Over 120 pharmaceutical products currently in use are derived from
plants. Mainly from tropical forest species (e.g. Taxol from Yew trees)
2. Plant-made pharmaceuticals (PMPs) and industrial products (PMIP)
(GE)

25. Plant Products:
Industrial products
• proteins
• enzymes
• modified starches
• fats
• oils
• waxes
• plastics
Pharmaceuticals
• recombinant human proteins
• therapeutic proteins and
pharmaceutical intermediates
• antibodies (plantibodies)
• vaccines

26. Strategies for Biopharming:
1.Plant gene expression strategies
• Transient transformation
• adv. – quick and easy production
• disadv. – small amount of product, processing pblms
• Stable transformation
• adv. – use for producing large quantities of protein, stability and storage
• disadv – gene flow - outcrossing w/native species
• Chloroplast transformation
• adv. – reduce gene flow through pollen
• disadv. – protein not stable for long periods of time therefore complications
w/extraction/processing times

27. Strategies for Biopharming:
2. Location of transgene expression
Protein quantity and preservation
• Whole plant
• adv. - an obtain large am ts of protein
• disadv. - problems w/preservation
• examples - tobacco, alfalfa, duckweed
• Target specific tissues (e.g. seed, root)
• adv. - high amts of protein in seed/root, long-term storage capability.
• examples: soy, corn, rice, barley

28. Strategies for Biopharming:
3. Selection of plant species and characteristics
• Mode of reproduction – self/outcrossing
• Yield, harvest, production, processing

29. Why use plants?
Advantages
Cost reduction
- scalability (e.g. Enbrel® )
- low/no inputs
- low capital cost
Stability
- storage
Safety
- eukaryotic production system
- free of animal viruses (e.g. BSE)
Disadvantages
Environment contamination
- gene flow
- wildlife exposure
Food supply contamination
-mistaken/intentional mixing
w/human food
Health safety concerns
-Variable, case-specific

30. Industrial products on the market:
Avidin by Sigma
• transgenic corn
• traditionally isolated from chicken egg whites
• used in medical diagnostics
GUS (b-glycuronidase) by Sigma
• transgenic corn
• traditionally isolated from bacterial
sources (E.Coli)
• used as visual marker in research labs
Trypsin by Sigma
• transgenic corn
• traditionally isolated from bovine pancreas
• variety of applications, including biopharmaceutical processing
• first large scale transgenic plant product
• Worldwide market = US$120 million in 2004

31. Plant-made Pharmaceuticals (PMPs):
• Plant- made vaccines (edible vaccines)
• Plant-made antibodies (plantibodies)
• Plant-made therapeutic proteins and intermediates
Unlike PMIPs, no PMPS are currently available on the market

32. Edible vaccines:
Advantages:
Administered directly
• no purification required
• no hazards assoc. w/injections
Production
• may be grown locally, where needed most
• no transportation costs
Naturally stored
• no need for refrigeration or special storage

33. Examples of edible vaccines under development:
• pig vaccine in corn
• HIV-suppressing protein in spinach
• human vaccine for hepatitis B in potato

34. Plantibodies:
• Plantibodies - monoclonal antibodies produced in plants
• Plants used include tobacco, corn, potatoes, soy, alfalfa, and rice
• Free from potential contamination of mammalian viruses
• Examples: cancer, dental caries, herpes simplex virus, respiratory
syncytial virus

35. Therapeutic proteins and intermediates:
• Blood substitutes – human hemoglobin
• Proteins to treat diseases such as CF, HIV, Hypertension, Hepatitis
B…..many others

36. Risks and Concerns:
Environment contamination
• Gene flow via pollen
• Non-target species near field sites e.g. butterflies, bees, etc
Food supply contamination
• Accident, intentional, gene flow
Health safety concerns
• Non-target organ responses
• Side-effects
• Allergenicity

37. Biopharm Opposition:
Main concern is containment.
Opponents want:
• a guarantee of 0% contamination of the food supply.
• full disclosure of field trials, crop, gene, location, etc.
• an extensive regulatory framework

38. Suggested Safeguards for biopharm
operations:
1. Physical differences
• e.g. “purple” maize, GFP
2. Sterility
• male sterile plants
• terminator technology
3. Easily detectable by addition of ‘reporter genes’
• e.g. PCR markers

39. Alternatives to biopharming?
Use only traditional drug production systems
• microbial, yeast and fungi
• mammalian cell culture
Use only fully contained production systems:
• plant cell cultures
• hydroponics (rhizosecretion)
• greenhouses
Use non-food crops
• tobacco
• hemp/cannabis

40. Future directions for agricultural
biotechnology:
Science has developed genetically enhanced crops and has/can develop
plant-made industrial and pharmaceuticals crops.
The extent to which these crops will be further developed for
commercial and/or humanitarian use will ultimately depend on…..
Public perception of
risk
Regulation

41. DNA Plasmid in Plant
Biotechnology

42. Importance of Plasmids:
• Plasmids are relatively small DNA sequences that can self replicate
• Exist independent
• Plasmids often carry antibiotic resistance genes that makes them
selectable
• They can be genetically modified - cut at specific locations using
restriction enzymes, and new DNA sequences included

43. Plasmids in plant biotechnology:
• In plant biotechnology recombinant plasmid vectors carrying genes
from other species with plant promoters have been used to
successfully transform plants so that they produce the foreign protein
• You may have heard of Genetically Modified Organisms (GMO)
• Everything from resistance to herbicides, insect pathogens, over
production of certain chemicals, modified fruit, the list goes on, have
been achieved.

44. Agrobacterium Tumefaciens:
• A single strand of the T-DNA complex is
transferred, along with the vir genes, to
the plant cell nucleus, where it is
inserted and expressed.
• The T-DNA of A. tumefaciens contains
genes encoding enzymes related to the
biosynthesis of cytokinin and auxins,
which cause the uncontrolled production
of transformed cells, leading to formation

45. Procedures of transferring plasmids:
Ti plasmid
Biolistic
Antisense technology
1
2
3

46. Ti plasmid:

47. Biolistic:
• Transformation by Gene Gun
Method.
• Blast tiny metal beads coated
with DNA into an embryonic
plant cell.

48. Antisense technology:
• Flavr SavrTM tomato
introduced in 1994
• Scientists isolated the PG gene,
produced a complementary
gene which produces a
complementary mRNA that
binds to the normal mRNA
inactivating the normal mRNA
for this enzyme
This is because ripe tomatoes normally produce the
enzyme polygalacturonase (PG), a chemical substance
that digests pectin in the wall of the plant

49. • Plant Biotechnology will save the
hunger people
• Soon it will be possible ti grow corn
in dry conditions without worrying
about insect and pests as well as
growing and maturing at an
astounding rate making it possible to
get more than one crop per season
Why we need it ????
The Future: From Pharmaceuticals
to Fuel
Herbicide resistance
Enhanced Nutrition

50. Scope of Plants Biotechnology In Pakistan:
Plants Biotechnology In Food and Agriculture.
• There are TWO types of food
• Fermented (pickles, yoghurt)
• Malted (powered milk, wheat)
• These food are produces by using bacteria and micro-organism
through Biotechnology.
• The development and release of the commercial transgenic crops is the
widely application of plant Biotechnology.
• It insert gene, make plant body strong that protect them from
herbicides attack and infection. As a result better yield is produced.

51. Plant biotechnology in Medicine.
• By using plant biotechnology, we make different medicines such as
interferons, antibodies and insulin etc.
Horticultural Technique.
• Plant biotechnology offers a vast application in horticulture.
• Horticultural scientists study crops, that are used for food, drugs or
aesthetics.
• In conservation of germplasm, virus-cleaning, biofertilizers and
biopesticides biotechnology, biotechnology covers a vast area in
Horticultural crops.

52. Tissue Culture.
• Tissue culture is the growth of tissues or cell separate from the
organisms.
• The application of cells, tissue and organ cultures central to many
modern crop-improvement programs.
Genetic Engineering of Plants.
• Many useful gene have been introduced into many plants from which
transgenic plants have been developed in which foreign DNA has been
integrated and resulted in the synthesis of appropriate gene product.