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Plant Cell Reports

, Volume 20, Issue 2, pp 150–156 | Cite as

High-efficiency gene transfer to recalcitrant plants by Agrobacterium tumefaciens

  • J. Ke
  • R. Khan
  • T. Johnson
  • D. A. Somers
  • A. Das
GENETIC TRANSFORMATION AND HYBRIDIZATION

Abstract

Agrobacterium tumefaciens efficiently transforms most plants. A few dicotyledonous plants and most monocotyledonous plants are, however, recalcitrant to A. tumefaciens infection. We investigated whether the constitutive synthesis of a high level of the T-strand DNA intermediate can improve the transformation efficiency of plants. We previously described a mutation in the vir gene regulator virG, virGN54D, that allows constitutive expression of the vir genes. We also described the isolation of a mutant plasmid that is present at a significantly high level in A. tumefaciens. The two mutations were combined to produce an A. tumefaciens strain that synthesizes a high level of T-strand DNA in an inducer-independent manner. DNA transfer efficiency of the mutant was measured by monitoring β-glucuronidase (GUS) expression in a transient transfer assay. A significant increase in the efficiency of DNA transfer to both rice and soybean was observed with the double mutant. The presence of virGN54D had a major positive effect on transformation efficiency.

KeywordsAgrobacterium tumefaciens Plant transformation Rice Soybean GUS gene expression 

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Copyright information

© Springer-Verlag Berlin Heidelberg 2001

Authors and Affiliations

  • J. Ke
    • 1
  • R. Khan
    • 2
  • T. Johnson
    • 1
  • D. A. Somers
    • 3
  • A. Das
    • 1
  1. 1.Department of Biochemistry, Molecular Biology and Biophysics and Plant Molecular Genetics Inst., University of Minnesota, 1479 Gortner Ave, St. Paul, MN 55108, USA e-mail: anath@cbs.umn.edu Fax: +1-612-6255780US
  2. 2.Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108, USAUS
  3. 3.Department of Agronomy and Plant Genetics, and Plant Molecular Genetics Institute, University of Minnesota, St. Paul, MN 55108, USAUS

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