Transformation of Botrytis cinerea with the hygromycin B resistance gene, hph.
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), conferring 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 micrograms 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.
Laboratoire de biochimie et de pathologie végétales, UPMC, Paris, France.
W
W
Hamada
P
P
Reignault
G
G
Bompeix
M
M
Boccara
D004352:Drug Resistance, Microbial / Q000235:genetics; D005800:Genes, Fungal; D005819:Genetic Markers; D006921:Hygromycin B / Q000494:pharmacology*; D003904:Mitosporic Fungi / Q000235:genetics*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011523:Protoplasts; D014170:Transformation, Genetic
D005819:Genetic Markers; D006921:Hygromycin B; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase