Vegetos- An International Journal of Plant Research
  • Year: 2018
  • Volume: 31
  • Issue: 4

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

  • Author:
  • Pankaj Borgohain1, Bhaben Chowardhara1, Bedabrata Saha1, Jay Prakash Awasthi1, Sibashish Kitaniya1, Lingaraj Sahoo2, Sanjib Kumar Panda1,
  • Total Page Count: 6
  • Page Number: 62 to 67

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.

Keywords

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