Effect of Acetosyringone and Callus Age on Transformation for Scutellum –derived Callus of Rice

An efficient and reproducible protocol for Agrobacteriummediated transformation of an agronomically useful Oryza sativa variety, Pusa Basmati1 (IET10364) has been standardized. Although several reporters have been claimed the Agrobacteriummediated transformation but here we studied the effect of acetosyringone and age of callus on transformation frequency for scutellumderived callus of Pusa basmati1. By optimizing these two parameters for Agrobateriummediated transformation, we can produce an efficient and reproducible protocol. The callus derived from mature rice seeds were used as explants for transformation studies. Callus were co-cultured with Agrobacterium tumefaciens EHA 105 harboring a binary vector p35SGUSINT that carried Betaglucuronidase (uid A) and It contains gene npt II, determining resistance to kanamycin, under the control of the constitutive (nos) promoter. Maximum Agrobacteriummediated transformation frequency was observed at 350μM/l acetosyringone with 35 days old callus.