African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12369

Full Length Research Paper

Agrobacterium mediated genetic transformation of popular Indica rice Ratna (IET 1411)

Arpita Basu¥
  • Arpita BasuÂ¥
  • Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700019, India.
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Sarmishtha Ray¥
  • Sarmishtha RayÂ¥
  • Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700019, India.
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Swadesh Sarkar
  • Swadesh Sarkar
  • Department of Botany, Gurudas College, 1/1, Suren Sarkar Road, Phoolbagan, Beliaghata, Kolkata 700010, India
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Tathagata Ray Chaudhuri
  • Tathagata Ray Chaudhuri
  • Department of Botany, Ashutosh College, 92 S.P. Mukherjee Road, Kolkata 700026, India.
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Surekha Kundu*
  • Surekha Kundu*
  • Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700019, India.
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  •  Received: 27 January 2014
  •  Accepted: 16 June 2014
  •  Published: 30 July 2014

Abstract

This is the first report of genetic transformation of indica rice cultivar Ratna (IET 1411). Agrobacterium tumefaciens strain LBA 4404 carrying a binary vector pCAMBIA1301 harboring the CaMV35S promoter driven modified GUS gene was used. Various parameters critical to rice transformation were optimized including callus induction medium, bacterial concentration, co-cultivation conditions, concentration of the plant  growth regulator 2,4-D and the concentration of acetosyringone. The transformed lines were analyzed using PCR for marker sequence and gusA gene expression by GUS activity assays. The regeneration and transformation frequency were calculated to be 70 and 47% respectively. This is a reproducible high efficiency transformation technique for the Indica rice cultivar Ratna (IET 1411).

 

Key words: Indica rice, Agrobacterium tumefaciens, callus induction, transformation, regeneration, GUS.

Abbreviation

Abbreviations: MS, Murashige and Skoog, 2,4-D, 2,4-dichlorophenoxyacetic acid, CH, casein hydrolysate.