Transformation of Botrytis cinerea with the hygromycin B resistance gene,hph |
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Authors: | W Hamada Ph Reignault G Bompeix M Boccara |
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Institution: | (1) Laboratoire de biochimie et de pathologie végétales, UPMC, 4, place Jussieu, C155, F-75252 Paris cedex 05, France;(2) Laboratoire de pathologie végétale, INA Paris, 16, rue Claude Bernard, F-75231 Paris cedex 05, France |
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Abstract: | A transformation method has been developed for the phytopathogenic fungus Botrytis cinerea. Protoplasts were transformed with pAN7-1 plasmid carrying the Escherichia coli hygromycin phosphotransferase gene (hph), confering hygromycin B resistance, downstream from an Aspergillus nidulans promoter. Molecular analysis showed that transformation resulted in an integration of the plasmid into different regions of the B. cinerea genome and occurred through non-homologous recombination. The frequency was 2–10 transformants per g of DNA. Transformants expressed phosphotransferase activity confirming that the hph gene conferred the hygromycin-resistance phenotype. All transformants analysed so far proved to be stable after several subcultures without any selective pressure. |
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Keywords: | Botrytis cinerea Hygromycin resistance Non-homologous recombination Phytopathogenic fungus |
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