Developing a highly efficient and reproducible plant regeneration protocol holds paramount significance in advancing technology for the genetic transformation of rice genotypes, facilitating their further utilization in crop improvement and breeding programs. Nagina 22 (N22), an aus type rice genotype, has tolerance against multiple stresses, requires very efficient transformation and regeneration protocol, for developing improved lines. However, a standardized regeneration protocol for this elite genotype has not been reported so far, limiting the use of this genotype as a background for genetic transformation. In the present study, for the first time, we are reporting a highly efficient transformation and regeneration protocol for the climate resilient N22. Mature seeds were used as explants for callus induction with varying concentrations (1-4 mg/L) of 2,4-dichlorophenoxyacetic acid (2.4-D), among which 3 mg/L of 2,4-D resulted in an impressive ~94% callus induction efficiency. Following this, the regeneration and resuspension media optimization is also carried out. Subsequent steps involved culturing embryogenic calli on media with different concentrations (2-6 mg/L) of 6-benzylaminopurine (BAP), in combination with 0.02 mg/L naphthalene acetic acid (NAA), to optimize the regeneration protocol. Media consisting of 5 mg/L BAP and 0.02 mg/L NAA exhibited a regeneration frequency of ~92% for the plantlets derived through non-transformed callus. Furthermore, our study revealed that reducing the salt concentration in the resuspension media resulted a notable enhancement in transformation efficiency of 44%. This in-vitro regeneration protocol provides valuable insights for future research and practical applications in genetic transformation of this elite rice genotype for various purposes.