Steps for Agrobacterium mediated transformation and regeneration of rice calli. Four indica rice cultivars i.e. IR64, PB1, CSR10 and Swarna were used here. Steps a-f have to be performed under dark conditions. (a) Callus initiation on callus induction medium. (b) Agro-infection and co-cultivation of calli with Agrobacterium tumefaciens. (c, d & e) First, second and third selection cycles of transformed calli/microcalli in presence of hygromycin (50 mg/l). (f) Morphology of microcalli after first phase of regeneration. Inset shows somatic embryo for each of the rice cultivar. (g) Regeneration of large number of shoots from microcalli during second phase of regeneration under light condition.