Developing an efficient and reproducible plant transformation protocol relies on callus induction and plant regeneration, which is prerequisite for genetic enhancement of crops, especially rice. The present study has been carried out in order to establish a genotype-independent regeneration and biolistic transformation protocol for rice varieties. Putative transgenic rice lines were confirmed by PCR analysis, DNA sequencing, and Southern analysis. The transformation protocol reported here is relatively simple and consistent and can be exploited in future biotechnological investigations particularly for gene transformation studies.