Pankaj Borgohain, Bhaben Chowardhara, Bedabrata Saha, Jay Prakash Awasthi, Sibashish Kitaniya, Lingaraj Sahoo, Sanjib Kumar Panda*
Keywords:
Agrobacterium tumefaciens, Plant transformation, tomato, hygromycin phosphotransferase (HPT), ZeBeta
An efficient Agrobacterium tumefaciens mediated transformation scheme was constituted using tomato cotyledonary leaf explants. 8 day old tomato seedling were used for excision of the explants. The two day pre-cultured explants were co-cultivated with Agrobacterium tumefaciens strain EHA105 harbouring the plant binary vector pCXSN, for 2 days in dark condition. pCXSN contains hygromycin phosphotransferase (HPT) as selectable marker and belongs to ZeBata series of binary vectors. Full strength MS media added on with 2µM zeatin, 0.1 µM IBA, 25 mg/L hygromycin and 250mg/L cefotaxime was optimized for multiple shoot regeneration and finally rooted in media containing IBA. The transgenic events were confirmed by genomic DNA PCR and transcript abundance was checked by semi-quantitative PCR.
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