Agrobacterium tumefaciens mediated genetic transformation of Ocimum gratissimum: A medicinally important crop

The present report focuses on the development of an efficient protocol for direct regeneration and Agrobacterium mediated genetic transformation of O. gratissimum. CN; when inoculated on MS + 1.5 mg/L + 0.5 mg/L + 0.5 mg/L KN medium has shown the best response with a regeneration frequency of 98% ± 0.4. A. tumefaciens LBA4404 containing binary vector pBI121, harboring gus-A reporter gene with intron under the transcriptional control of Cauliflower Mosaic Virus (CaMV) 35S promoter and neomycin phosphotransferase (npt-II) as selection marker genes, to achieve maximum transformation frequency (20% ± 0.7). To carry out transformation, parameters like O.D., infection duration, concentration of AS, surfactant, (O.D600-0.6, 15 min, 300 μM, 0.01%) were standardized. The putative transformants were screened on SMM having 50 mg/L kanamycin and subsequently rooted on ½ MS. The confirmation was done through PCR. The established regeneration and transformation protocol could be used as a tool to elucidate the functional genomics of important/novel compound which can be used in the preparation of therapeutic formulations for future.