Research Article | Published: 11 January, 2019
First Page: 62
Last Page: 67
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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Pankaj Borgohain Plant Molecular Biotechnology Laboratory, Department of Life Science and Bioinformatics, Assam University, Silchar 788011, INDIA Bhaben Chowardhara Plant Molecular Biotechnology Laboratory, Department of Life Science and Bioinformatics, Assam University, Silchar 788011, INDIA Bedabrata Saha Plant Molecular Biotechnology Laboratory, Department of Life Science and Bioinformatics, Assam University, Silchar 788011, INDIA Jay Prakash Awasthi Plant Molecular Biotechnology Laboratory, Department of Life Science and Bioinformatics, Assam University, Silchar 788011, INDIA Sibashish Kitaniya Plant Molecular Biotechnology Laboratory, Department of Life Science and Bioinformatics, Assam University, Silchar 788011, INDIA Lingaraj Sahoo Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati 781039, INDIA Sanjib Kumar Panda* Plant Molecular Biotechnology Laboratory, Department of Life Science and Bioinformatics, Assam University, Silchar 788011, Indiadrskpanda@gmail.com