This document provides an overview of transgenesis and the development of transgenic plants. It defines key terms like transgene and transgenic plants. It describes traditional plant breeding techniques and compares them to transgenic technology. Transgenic technology allows genes to be transferred between any organisms, while traditional breeding is limited to the same genus. Reasons for developing transgenic plants include crop improvement, disease resistance, and stress tolerance. The document outlines the process of developing transgenic plants, including vector-mediated gene transfer using Agrobacterium and biolistic methods. It provides examples of transgenic plants created for insect resistance, herbicide tolerance, drought tolerance, and more. Both advantages and disadvantages of transgenic plants are discussed.

1. TRANSGENESIS AND THE DEVELOPMENT OF
TRANSGENIC PLANTS
COURSE TUTOR: PROF. DR. SMARANDA VANTU
BY
AHMAD ALI
ERASMUS STUDENT
APPLICATION OF GENETIC ENGINEERING
INDUSTRIAL BIOTECHNOLOGY
Department of Molecular Genetics, Faculty of Biology
University of Alexandra Ioan Cuza
Romania

2. TERMS TO KNOW
 Transgene : It is a gene or genetic material that has been
transferred naturally or by any of a number of genetic engineering
techniques from one organism to another.
 Transgenesis : The process of introducing an exogenous gene
called a transgene into a living organisms so that the organism
will exhibit a new property and transmit that property to its
offspring.
 Transgenic Plants : The plants which expresses the characters
coded by the transgene are called Transgenic plants.

3. TRADITIONAL BREEDING
 Selective Breeding used in the History
genetics studies started with Mendel
 Cross pollination : Pollen from one
plant to stigma of another plant.
 Found dominate characteristics in
plants.
 Disadvantages of Traditional
breeding:
1. Long process
2. Lot of man power
3. Limited possibility of improved traits.

4. TRANGENIC TECHNOLOGY V/S TRADITIONAL
BREEDING:
 Trangenic Technology : Transform gene from
any source.
Eg: animals, bacteria, virus etc
 Traditional Breeding : Move genes only
between members of a particular genus of
plants.

5. WHY TRANSGENIC PLANTS?
Why gene transfer?
 Crop improvement
 Disease resistance
 Stress tolerance
 Improved performance
 Value-added traits
Basic studies
 Gene expression
 Reverse genetics - understanding functioning of
unknown genes
 Biochemistry and metabolism

6. TRANGENIC PLANTS
 The plants whose genome is altered by adding one or
more transgenes are known as transgenic plants .
 The Phenomenon of introduction of exogenous DNA into
the genome to create and maintain a stable and
heritable character.
History of GMO’s
 1982-1st transgenic plant produced which is an antibiotic
resistance tobacco plant.
 1984-1st successful plant genetic engineering
experiments was done by using Caulimovirus vector.
 1994-1st genetically modified crop approved for sale in
US was Flavr-Savr tomato.

7. DEVELOPMENT OF TRANSGENIC PLANTS
 The basic requirements of transformation are,
1. A target genome.
2. A vector to carry the gene.
3. Modification of the foreign DNA to increase the level of
gene expression.
4. Method to deliver the plasmid DNA into the cell.
5. Methodology to identify the transformed cell.
6. Tissue culture to recover the viable plants from the
transformed cells.

8. TRANSGENIC PLANTS HAVE BEEN PRODUCED BY
ADDITION OF ONE OR MORE FOLLOWING GENES
 Endo-toxin gene from Bt
 Protease inhibitors
 Alpha amylase inhibitors
 Lectins
 Enzymes
 Pyramiding genes
 Gene transfer strategies: Systems and vector
1.Vector mediated gene transfer
Agrobacterium mediated gene transfer
2.Direct (or) vectorless DNA transfer
Gene gun, Electroporation ,Microinjection

9. AGROBACTERIUM MEDIATED GENE TRANSFER
 Natural soil borne bacterium – Crown gall disease in plants .
 It is an efficient vector of DNA because it has the ability to transmit a
fragment of its large plasmid into the nuclear genome of an infected
cell .
 The fragment called T-DNA, transfer contained genes- Oncogenes,
that induce tumors in the plant tissues.
 Instead of Oncogenes, the transgenic Agrobacterium is then used to
transfer the delta endotoxin protein into the plant cell chromosomes .
 Plant cell is cultured and grown into a whole plant whose ells
contains the toxic protein .
 These transgenic resistant plants then produce seed expressing the
insect resistant trait, which can be commercialized.
 This procedure has been used to transfer Bt delta endotoxin gene to
many plants including (Cotton, tobacco, tomato and potato).

10. F.C STEWARD EXPERIMENT WITH CARROTS:

11. ESSENTIAL ELEMENTS FOR CARRYING A TRANSGENE ON
TI PLASMIDS

12. Agrobacterium carrying a Ti plasmid is added to plant tissue growing in
culture. The T-DNA carries an antibiotic resistance gene (neomycin in this
figure) to allow selection of successfully transformed plant cells. Both
callus cultures (A) and liquid cultures (B) may be used in this procedure

13. BIOLISTIC METHOD (PARTICLE GUN)
 Also called as “Gene gun or Micro-projectile bombardment”
method.
 DNA is bound to tiny particles of gold or tungsten which are
shot into plant tissue.
 The particles penetrate both the cell wall and membranes.
 DNA separates from the metal and is integrated into the
plant genome inside the nucleus.
 Successfully for monocots- Wheat, maize.
 Disadvantage-
 Serious damage to the cellular tissue.

15. APPLICATIONS OF TRANSGENESIS
Edible Vaccine:
 The genes encoding antigens of bacterial and viral
pathogens can be expressed in plants in a form in which
they retain native immunogenic properties.
 In the edible vaccine, transgenic plants are used as
vaccine production systems.
Vaccines can be:
 PROPHYLACTIC : A prophylactic is a medication or a treatment designed
and used to prevent a disease from occurring.
For example, prophylactic antibiotics may be used before a rheumatic
fever to prevent the subsequent development of Sydenham's chorea.
 THERAPEUTIC: A therapeutic vaccine is one in which the vaccine is used
after infection occurs, aiming to induce anti-viral immunity to alter the course
of disease.

16. DEVELOPING AN EDIBLE VACCINE
Two ways
1. In one case , the entire structural gene is inserted into
plant transformation vector between 5‟ and 3‟
regulatory element ; this will allow the transcription and
accumulation of encoding sequence in the plant.
2. In the second case , epitope within the antigen are
identified ,DNA fragment encoding these can be used
to construct gene by fusion with a coat protein gene
from plant virus e.g. TMV or CMV .

17. CONT’D

18. MECHANISM OF ACTION:
 The goal of oral vaccination is to stimulate the mucosal and systemic
immunity against pathogen.
 Edible vaccine when taken orally undergoes the mastication process and
the majority of plant cell degradation occur in the intestine as a result of
action of digestive or bacterial enzyme on edible vaccine.
 Payer's patches (PP) are an enriched source of IgA producing plasma cells
and have the potential to populate mucosal tissue and serves as mucosal
immune effector site.
 The breakdown of edible vaccine near PP , consisting of the 30-40 lymphoid
nodules on the outer surface of intestine and contain follicles.
 These follicles act as the site from which antigen penetrates the intestinal
epithelium ,thereby accumulating antigen within organized lymphoid
structure .
 The antigen then comes in contact with M-cell .
 M cell passes the antigen to macrophages and B cell.
 These B cell activates the T cell to provide immune response .
 In this way the immunity is activated by the edible vaccine.

19. WILLAIM ET.,AL 2000

20. CASE STUDY: DESIGN OF A TYPE-1 DIABETES VACCINE CANDIDATE USING
EDIBLE PLANTS EXPRESSING A MAJOR AUTOANTIGEN

21. CURRENT STATUS: CLINICAL TRAILS
Hepatitis B (Mishra et.,al 2008).
 First human trials of a potato based vaccine against hepatitis B have
reported encouraging results.
 The amount of HBsAg needed for one dose could be achieved in a
single potato.
 When cloned into CaMv , plasmid HBsAg subtype showed higher
expression in roots as compared to leaf tissue of the transgenic
potato.

22. 2.TRANSGENIC PLANT AS A BIOREACTOR:
 A bioreactor is basically a device in which the organism (cells) are
cultivated and motivated to from the desired products.
 Transgenic plants have significant potential in the bioproduction of
complex human therapeutic agents due to ease of genetic
manipulation, lack of potential contamination with human pathogen.
Types of Plant Bioreactor:
1).Plant suspension culture 2). Chloroplast bioreactor
3). Hairy root system 4). Seed based plant bioreactor.

23. BIOPLASTICS
 Biodegradable plastics or bio-plastics are chemically poly hydroxyl
alkanets (PHAs).
 They are currently being produced in large quantities by microbial
fermentation.
Bioplastics: 1). PHB Formation in Plastid 2). PHB formation in
Cytoplasm.

24. GENETICALLY MODIFIED STRAWBERRIES
 Antifreeze protein gene sequence actually taken from the Antarctica
Fish called arctic flounder fish, protein are actually glycoprotein in
nature and are used to lower the freezing point of strawberries', as they
have a problem of freezing at the freezing condition.

25. TRANSGENIC PLANTS AGAINST BIOTIC AND ABIOTIC STRESS
Transgenic wheat against drought resistance:
 Transgenic wheat carrying a GmDREB1 gene from soybean, under a
ubiquitin promoter, showed drought tolerance.
 main interest is to improve wheat plants could grow well under water
stress.
1. One way is inserting genes for compatible osmolytes such as
sugar and amino acids.
2. The other strategy is constitutive over-expression of the late
embryogenic abundant proteins which provide dehydration
tolerance.

26. ADVANTAGES AND DISADVANTAGES OF TRANSGENICS PLANTS
 Improvement in nutritional value of food.
 Increase in farmers income.
 Increase in food supply.
 More convenient and flexible to use.
 Safer environment through decreased use of pesticides.
 Improved the quality of ground and surface water with less pesticide residues.
 Safe to non-target organisms and human beings.
 Disadvantages:
 Secondary pest incidence.
 Disruption of pollinators and plant communities would occur if the toxin is
expressed in plant nectar and pollen.
 Antibiotic resistance due to antibiotic markers
 GM ingredients cause cancer- Histopathologist (Dr. Stanley Ewan) “food
and water contaminated with GE material could increase the growth of
Malignant tumor.
 GM food could raise new allergy outbreak in humans – GM soybean
containing “Brazilian protein” was allergic to humans and was withdrawn
from production.

27. CONCLUSION:
 Transgenic plants have the potential to solve many of the
worlds hunger and malnutrition problems
 Help to protect and preserve the environment by
increasing yield and reducing reliance upon chemical
pesticides and herbicides.

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30. Thank you!
We Mature with mistake, not
year’s.