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Plant Molecular Biology Reporter

, Volume 23, Issue 1, pp 67–73 | Cite as

An improvedAgrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars

  • K. K. Kumar
  • S. Maruthasalam
  • M. Loganathan
  • D. Sudhakar
  • P. Balasubramanian
Protocols

Abstract

We report here a high-efficiency transformation protocol for recalcitrant indica rice cultivars IR64 and IR72 with the selectable marker genehph and thegusA reporter gene. Factors that favor high-efficiency transformation were found to be use of 2-month-old mature seed-derived embryogenic calli, maltose as a source of carbon, a higher concentration of 2,4-dichlorophenoxyacetic acid, and both phytagel and agar as gelling agents. The putative transgenic (T0) plants were analyzed for integration of the transgene through polymerase chain reaction and Southern blotting analyses. Various factors thought to be responsible for increased transformation efficiency are discussed.

Key words

Agrobacterium indica rice mature seed-derived calli rice transformation transgenic rice 

Abbreviations

2,4-D

2,4-dichlorophenoxyacetic acid

BAP

6-benzylaminopurine

MS

Murashige and Skoog

NAA

α-naphthaleneacetic acid

OD

optical density

PCR

polymerase chain reaction

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Copyright information

© International Society for Plant Molecular Biology 2005

Authors and Affiliations

  • K. K. Kumar
    • 1
  • S. Maruthasalam
    • 1
  • M. Loganathan
    • 1
  • D. Sudhakar
    • 1
  • P. Balasubramanian
    • 1
  1. 1.Rice Transformation Laboratory, Department of Plant Molecular Biology and Biotechnology, Center for Plant Molecular BiologyTamil Nadu Agricultural UniversityCoimbatoreIndia

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