Abstract
Ocimum gratissimum is renowned for its immense role in traditional, ayurvedic and unani system of medicines. The essential oil has a plethora of active ingredient like triterpenes, polyphenols and phenylpropanoids. The active ingredients claim most of the properties like anticancerous, anti HIV, antipyretic, antinociceptive, antiurolithiatic etc. Screening of biologically active ingredients from this valuable plant may provide us the basis to produce novel pharmaceutically important drugs and medicines. Considering the importance of this plant, it is the need to have a well developed regeneration and transformation protocol.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+1.5mg/L+0.5mg/L+0.5mg/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, 15min, 300μM, 0.01%) were standardized. The putative transformants were screened on SMM having 50mg/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.
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