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.
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Received: 4 August 2000 / Revision received: 9 October 2000 / Accepted: 12 October 2000
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Ke, J., Khan, R., Johnson, T. et al. High-efficiency gene transfer to recalcitrant plants by Agrobacterium tumefaciens. Plant Cell Reports 20, 150–156 (2001). https://doi.org/10.1007/s002990000271
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DOI: https://doi.org/10.1007/s002990000271