High-efficiency gene transfer to recalcitrant plants by Agrobacterium tumefaciens |
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| Authors: | J Ke R Khan T Johnson D A Somers A Das |
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| Institution: | (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: +1-612-6255780, US;(2) Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108, USA, US;(3) Department of Agronomy and Plant Genetics, and Plant Molecular Genetics Institute, University of Minnesota, St. Paul, MN 55108, USA, US |
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| 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.
Received: 4 August 2000 / Revision received: 9 October 2000 / Accepted: 12 October 2000 |
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| Keywords: | Agrobacterium tumefaciens Plant transformation Rice Soybean GUS gene expression |
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