Development of a binary vector system for plant transformation based on the supervirulent Agrobacterium tumefaciens strain Chry5 |
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| Authors: | R. S. Torisky L. Kovacs S. Avdiushko J. D. Newman A. G. Hunt G. B. Collins |
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| Affiliation: | (1) Department of Horticulture, North Carolina State University, Box 7609, Kilgore Hall, Raleigh, NC 27695-7609, USA, US;(2) State Fruit Experiment Station, Southwest Missouri State University, Mountain Grove, MO 65711, USA, US;(3) Department of Agronomy, University of Kentucky, N212 Agricultural Science Center North, Lexington, KY 40546-0091, USA Fax no.: +1-606-323-1952 E-mail: AGR132@ukcc.uky.edu, US;(4) Department of Biology, Lycoming College, 700 College Place, Williamsport, PA 17701-5192, USA, US |
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| Abstract: | This report describes the disarming of Agrobacterium tumefaciens Chry5, a strain highly tumorigenic on soybean. Disarming was achieved by removing an approximately 16.5-kb segment of the 285-kb Ti plasmid pTiChry5, including approximately 4 kb of the oncogenic T-DNA and an extended region right of the T-DNA, and replacing it with a gene for carbenicillin resistance, through homologous recombination. The deletion was confirmed with Southern analysis, and the loss of tumorigenicity was verified in tobacco and tomato plant stem inoculation assays. The deletion mutant, named KYRT1, successfully transferred the β-glucuronidase (GUS) gene into tobacco leaf tissue, producing GUS-expressing callus which could be regenerated into viable plants. In a comparative study, the transformation efficiency of A. tumefaciens KYRT1, GV3850, and EHA105 was assayed by inoculating cotyledonary node explants. The results of this study revealed that, in a binary vector system, KYRT1 is equally or more effective than EHA105 or GV3850 at delivering DNA into soybean. Received: 30 January 1997 / Revision received: 10 June 1997 / Accepted: 5 July 1997 |
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| Keywords: | Glycine max Agrobacterium tumefaciens Transformation Virulence Ti plasmid |
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