Establishment of a highly efficient Regeneration system in Tomato var. Pusa ruby amenable to Agrobacterium tumefaciens Mediated Plant Transformation system Borgohain Pankaj1, Chowardhara Bhaben1, Saha Bedabrata1, Awasthi Jay Prakash1, Kitaniya Sibashish1, Sahoo Lingaraj2, Panda Sanjib Kumar1,* 1Plant Molecular Biotechnology Laboratory Department of Life Science and Bioinformatics, Assam University, Silchar, 788011, India 2Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, 781039, India *Corresponding author: Plant Molecular Biotechnology Laboratory, Department of Life Science and Bioinformatics, Assam University, Silchar, 788011, India. Email: drskpanda@gmail.com
Online published on 11 January, 2019. Abstract An efficient A grobacterium 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 A grobacterium 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 semiquantitative PCR. Top Keywords Agrobacterium tumefaciens, Plant transformation, tomato, hygromycin phosphotransferase (HPT), ZeBeta. Top |