1. Transgenic rice and biosafety procedures D.S. Brar PGPB, IRRI Rice Production Course

2. IRRI: Rice Production Course Learning objectives To describe general methods for rice improvement To describe genetic improvement of rice To define and describe transgenic rice To describe a brief history of transgenics in rice To explain production, characterization and evaluation of transgenic rice

3. IRRI: Rice Production Course Option 1Management practices – Agronomic practices (sowing, fertilizer, water, soil preparation, etc.) – Integrated Pest Management (IPM) – Integrated nutrient management – Post harvest losses (drying, storage, etc.) Option 2Genetic improvement Option 3Combination of options 1 and 2 Can anyone describe methods for rice improvement?

4. IRRI: Rice Production Course What are the methods for genetic improvement of rice? 1.Conventional breeding methods – Hybridization, backcrossing, selection and evaluation of progenies 2.Wide hybridization (rice x wild species) 3.Tissue culture (anther culture, somatic cell culture)

5. IRRI: Rice Production Course 4. Mutation (create new variation) 5. Molecular breeding (use of molecular markers) 6. Transformation (genetic engineering) – Introducing transgene/foreign genes through recombinant DNA technique but not through sexual hybridization

6. IRRI: Rice Production Course Can anyone define “transgenic rice”?  Broadly speaking refers to any rice plant that carries foreign gene (s) introduced through genetic engineering techniques  The transgene could be from a micro-organisms and different plants

7. IRRI: Rice Production Course  Gene = introduced directly into cultured cells (callus) and/or organs (immature embryo) using biolistic or Agrobacterium methods of transformation  Rice plants are regenerated from transformed cells and progenies and tested for the desirable agronomic trait (s) (eg. stem borer resistance) Transgenic rice

8. IRRI: Rice Production Course  Transgenic rice = commonly referred as GM (genetically modified) rice  Transgenics are released into environment after series of biosafety tests  Biosafety = safety of living organisms (human, microorganisms, plants, animals & insects,…)  IRRI follows biosafety guidelines of NCBP, Philippines. Transgenic rice

9. IRRI: Rice Production Course

12. A schematic protocol for production of fertile transgenic plants using: biolistic systems, protoplast systems Agrobacterium systems (Datta,et al.)

13. IRRI: Rice Production Course Brief history of transgenics in rice  Transgenic rice first produced in 1988  Two methods commonly used to produce transgenics  Biolistic  Agrobacterium mediated  Both indica and japonica varieties have been transformed

14. IRRI: Rice Production Course  Transgenic rice have been produced carrying genes for:  herbicide resistance, bacterial blight, virus,  sheath blight resistance  stem borer resistance  high iron, golden rice  No commercial release of transgenic rice

15. IRRI: Rice Production Course Transgenic rice – production, characterization and evaluation 1.Transgene availability – local or imported 2.Insertion into the elite line 3.Growing of transgenics in containment facility (CL4), low level containment facility (screen house) 4.Characterization and evaluation of transgenics (based on agronomic and molecular chracteristics) 5.Commercial release of transgenics

16. IRRI: Rice Production Course Transgenic rice with Xa21 gene showing resistance to bacterial blight

17. IRRI: Rice Production Course Transgenic rice (golden rice) with psy, crtl, lcy genes possessing B-carotene

18. IRRI: Rice Production Course Transgenic rice with Bt gene showing resistance to stem borer

19. IRRI: Rice Production Course Some examples of transgenic rice produced at IRRI TraitTransgene(s)Status Bacterial blight Xa21 Field tested in India, China and Philippines Stem borer resistance Bt (cry1A, 1B, 1C, etc) Field tested in China. Bt genes introduced into many cultivars Sheath blight resistance Chitinase Transgenics show increased tolerance to sheath blight Abiotic stress tolerance DREB Transgenics produced are under evaluation Golden rice (ß-carotene) psy, cry1, lcy Many varieties transformed. Backcrossing used to introduce into other varieties by NARES

20. IRRI: Rice Production Course Transgenic rice and biosafety Biosafety componentActivity (ties) Food and health safety Assessment of GM rice for toxicity, allergenicity and nutritional imbalance Environmental safety Effect of GM rice on: beneficial insects soil micro-organisms non-target species (plants, animals, microbes) Gene flow from transgenic to non-transgenic rice and wild species of Oryza Development of resistance to pests Capacity building on biosafety of GM rice Training courses/workshop on risk assessment, regulatory processes and public awareness Detection of transgene(s) using PCR kits

21. IRRI: Rice Production Course

23. Can anyone briefly summarize the importance of transgenic rice? Questions or comments?