Optimization of Callus Induction in Rice Genotype For Novel Gene transformation

Abstract

Rice, one of the most important cereal crop directly consumed by human and second largest crop in panting acreage after wheat in the world. To meet the growing demand for rice, biotechnological intercession for its improvement become increasingly important. Such intercession hinge on the development of efficient transformation protocol. However, production of callus and its subsequent regeneration are the limiting step in crop plant to be manipulated by biotechnological mean. In present studies, callus induction and regeneration protocol was optimized for the varieties Pusa Basmati-1, Pusa Sundha-4, Pant Dhan- 10 and saket-4. The media containing MS and N6 salt, carbohydrate source, amino acid and growth regulator were evaluated. Medium N6 gave higher callus induction frequency than MS and for regeneration MS medium proved better. While the variety, Pusa Basmati-1 showed higher callus induction response than the other varieties studied. Calli from Pusa Basmati-1, Pusa Sundha-4 were transformed through Agrobacterium tumefaciens strain EHA 105 harboring plasmid pCAMBIA 1305 that carries gene for B-glucuronidase and hygromycin phosphotransferase (hpt) which gave better transformation efficiency.