GENETIC ENGENEERING IN AGRICULTURE

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1. WELCOME

2. PRESENTED BY GANESH H S DEPARTMENT OF PLANT PATHOLOGY

3. PRESENTATION ON “GENETIC ENGINEERING FOR DISEASE RESISTANT”

4. AREAS OF PRESENTATION INTRODUCTION GENETIC ENGEENERING HISTORY METHODS OF GENE TRANSFER MECHANISAM CHALLENGES CASE STUDY

5. Methods for plant genome manipulation Classical breeding Transgenic approach Targeted genome editing An alternative to both classical plant breeding and transgenic approach 5

6. WHAT IS GENETIC ENGEENERING? “ Genetic engineering refers to the direct manipulation of DNA to alter an organism’s characteristics (phenotype) in a particular way .”   Genetic engineering, sometimes called genetic modification, is the process of altering the  DNA   in an organism’s  genome It may also mean extracting DNA from another organism’s genome and combining it with the DNA of that individual.

7. HISTORY 1991- Dewit et al., reported first fungal Avirulence gene Avr-9 from Cladosporium fulvum 1992- Johal and Briggs reported first R gene HM-1 in maize, resistant to Cochliobolus carbonum race-1 1994 – Flavr Savr , transgenic tomato in USA (Anil and deepak , 2001 )

8. Country GM planted area (million hectares) Biotech crops United states 73.1 Maize, Soybean, Cotton, Canola, Sugarbeet, Alfalfa, Papaya and Squash Brazil 42.2 Soybean, Maize and Cotton Argentina 24.3 Soybean, Maize and Cotton India 11.6 Cotton Canada 10.8 Canola, Maize, Soybean and Sugar beet (ISAAA,2014) AREA COVERED BY GM CROPS IN WORLD

9. APPLICATION OF GENETIC ENGENEERING Insect resistance Virus resistance Fungal/bacterial disease resistance Herbicide resistance Seed storage proteins Biochemical production and Edible vaccines. Nutrition enhancement Abiotic stress tolerance

10. TRANSGENEIC CROP “The plants in which foreign genes have been introduced through genetic engineering are called transgenic plants”

11. METHODS FOR GENE TRANSFER Isolate and clone gene of interest Add DNA segments to initiate gene expression Add selectable markers Transformation Select transformed cells or tissues Regenerate whole plants

12. 1. Identify and clone the gene of interest The most limiting step in the transgenic process. Public and private labs are directing more efforts to locate, identify, characterize, and clone genes of agricultural importance. ( Bhat and Chopra,2005)

13. 2. Add DNA segment to initiate gene expression Promoter : CaMV 35S promoter Terminator sequence: Nopaline synthase ( nos ) transcription terminator sequence from Agrobacterium tumefaciens . Promoter Termination sequence Gene of interest ( Bhat and Chopra, 2005)

14. Gene transfer marker is needed to identify cells with the new genes. Typically antibiotic resistance genes are used as markers. Antibiotic resistance gene Promoter Gene of interest Termination sequence 3. Add selectable marker ( Bhat and Chopra, 2005)

15. 4. Gene transfer Vector mediated Micro injection Electroporation Gene gun

16. VECTOR MEDIATED GENE TRANSFER Agrobacterium –mediated gene transfer

17. Ti plasmid (Wani,2010)

18. VECTOR LESS GENE TRANSFER Direct uptake of DNA. Electroporation . Microinjection. Gene gun.

19. PROPERTIES OF GOOD HOST Easy to transfer. Support the replication of recombinant DNA. Free from elements which interfere in the replication of recombinant DNA. Lacks active restriction enzymes . Does not have methylases . 21 ( Bhat and Chopra,2005)

20. MECHANISM OF TRANSGENEIC PLANTS IN DISEASE RESISTANCE Directly toxic to or which reduce growth of the pathogen. Destroy or neutralize of the pathogen components. Potentially enhance the structural defenses in the plant. Release signals that can regulate plant defenses . Resistance gene(R) products involved in R/ Avr interactions and Hypersensitive response ( Punja , 2008)

21. ‘ R’ genes Gene Host pathogen Hm1 corn Cochliobolus carbonum Pto Tomato Pseudomonas syringae pv . tomato RPS2 Arabidopsis P. syringae pv . Tomato, P. syringae pv . maculicola N Tobacco Tobacco Mosaic Virus Cf9 Tomato Cladosporium fulvum L 6 Linseed Melampsora lini

22. TRANSGENIC PLANT THAT TOLERATE ABIOTIC STRESSES CROP GENE FOR AGANIST Brinjal Mannitol phsophodehydrogenase Osmatic stress Paddy Glutamine S transferase Lower temperature Tobacco Trehalose phosphate Drought

23. CASE STUDY 1

24. Percentage of susceptibility to infection for chitinase tomato R1 transgenic and control plants after transplanting into soil infested with F. oxysporum

25. CASE STUDY 2

26. Analysis of the tobacco transgenic plants with the challenge from Fusarium moniliforme

27. Analysis of lesion number and lesion diameter in control and transgenic plants after conidial spray in detached leaf assays.

28. CASE STUDY 3

29. Positive impacts Wider selection of traits for improvement It is faster and lower in cost Desired change can be achieved in very few generations Preater precision in selecting characteristics Reduces risk of random occurrence of undesirable traits ( Arunraj and Gajendra , 2001)

30. Challenges Technical Limitations It is unpredictable Transgene inactivation and silencing by the host system . Co-suppression Cytotoxic and inhibitory to host growth and metabolism. Lack of technique for transferring multiple genes by transformation ( Arunraj and Gajendra , 2001)

31. Social Limitations Toxicity New pathogenic bacteria. Contaminate native genetic information Resistance Allergies ( Arunraj and Gajendra , 2001)

32. conclusion

33. ANY QURIES??

34. THANK YOU Thank you