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Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii

Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii R. G. Upchurch 1 * , M. Ehrenshaft 1 , D. C. Walker 2 † and L. A. Sanders 2 ‡ 1 Agricultural Research Service, U.S. Department of Agriculture, and Department of Plant Pathology, 2 North Carolina State University, Raleigh, North Carolina 27695-7616 ABSTRACT An altered β-tubulin gene that confers resistance to the fungicide benomyl was isolated from a genomic library of a UV-induced mutant of Cercospora kikuchii and used as a selectable marker for transformation. The level of benomyl resistance conferred to the transformants was at least 150-fold greater than the intrinsic resistance of the C. kikuchii recipient protoplasts. In the majority of cases, the tubulin fragment was integrated at the native β-tubulin locus, apparently by gene replacement or gene conversion. The frequency of transformation ranged from 0.2 to 6 transformants per μg of DNA, depending on the recipient strain. Transformation with linearized plasmid resulted in a higher frequency, without changing the type of integration event. Transformants were phenotypically stable after eight consecutive transfers on medium without benomyl. This is the first report of a genetic transformation system for a Cercospora species. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied and Environmental Microbiology American Society For Microbiology

Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii

Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii

Applied and Environmental Microbiology , Volume 57 (10): 2935 – Oct 1, 1991

Abstract

Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii R. G. Upchurch 1 * , M. Ehrenshaft 1 , D. C. Walker 2 † and L. A. Sanders 2 ‡ 1 Agricultural Research Service, U.S. Department of Agriculture, and Department of Plant Pathology, 2 North Carolina State University, Raleigh, North Carolina 27695-7616 ABSTRACT An altered β-tubulin gene that confers resistance to the fungicide benomyl was isolated from a genomic library of a UV-induced mutant of Cercospora kikuchii and used as a selectable marker for transformation. The level of benomyl resistance conferred to the transformants was at least 150-fold greater than the intrinsic resistance of the C. kikuchii recipient protoplasts. In the majority of cases, the tubulin fragment was integrated at the native β-tubulin locus, apparently by gene replacement or gene conversion. The frequency of transformation ranged from 0.2 to 6 transformants per μg of DNA, depending on the recipient strain. Transformation with linearized plasmid resulted in a higher frequency, without changing the type of integration event. Transformants were phenotypically stable after eight consecutive transfers on medium without benomyl. This is the first report of a genetic transformation system for a Cercospora species.

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Publisher
American Society For Microbiology
Copyright
Copyright © 1991 by the American society for Microbiology.
ISSN
0099-2240
eISSN
1098-5336
Publisher site
See Article on Publisher Site

Abstract

Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii R. G. Upchurch 1 * , M. Ehrenshaft 1 , D. C. Walker 2 † and L. A. Sanders 2 ‡ 1 Agricultural Research Service, U.S. Department of Agriculture, and Department of Plant Pathology, 2 North Carolina State University, Raleigh, North Carolina 27695-7616 ABSTRACT An altered β-tubulin gene that confers resistance to the fungicide benomyl was isolated from a genomic library of a UV-induced mutant of Cercospora kikuchii and used as a selectable marker for transformation. The level of benomyl resistance conferred to the transformants was at least 150-fold greater than the intrinsic resistance of the C. kikuchii recipient protoplasts. In the majority of cases, the tubulin fragment was integrated at the native β-tubulin locus, apparently by gene replacement or gene conversion. The frequency of transformation ranged from 0.2 to 6 transformants per μg of DNA, depending on the recipient strain. Transformation with linearized plasmid resulted in a higher frequency, without changing the type of integration event. Transformants were phenotypically stable after eight consecutive transfers on medium without benomyl. This is the first report of a genetic transformation system for a Cercospora species.

Journal

Applied and Environmental MicrobiologyAmerican Society For Microbiology

Published: Oct 1, 1991

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