Establishment of a highly efficient Regeneration system in Tomato var. Pusa ruby amenable to Agrobacterium tumefaciens Mediated Plant Transformation system

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Research Article | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Website:www.vegetosindia.org
Pub Email: contact@vegetosindia.org
Doi: 10.5958/2229-4473.2018.00095.2
First Page: 62
Last Page: 67
Views: 1801


Keywords: Agrobacterium tumefaciens, Plant transformation, tomato, hygromycin phosphotransferase (HPT), ZeBeta


Abstract


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.

Agrobacterium tumefaciens, Plant transformation,  tomato,  hygromycin phosphotransferase (HPT), ZeBeta


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Acknowledgements



Author Information


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, India
drskpanda@gmail.com