title: Genetic analysis of fenhexamid-resistant field isolates of the phytopathogenic fungus Botrytis cinerea.
abstract: The hydroxyanilide fenhexamid, one of the latest antibotrytis fungicides, active especially against leotiomycete plant-pathogenic fungi, inhibits 3-ketoreductase of the C-4-demethylation enzyme complex during ergosterol biosynthesis. We isolated Botrytis cinerea strains resistant to various levels of fenhexamid from French and German vineyards. The sequence of the gene encoding 3-ketoreductase, erg27, varied according to levels of resistance. Highly resistant isolates, termed HydR3(+), all presented a modification of the phenylalanine at the C terminus of the putative transmembrane domain at position 412, either to serine (85% of the isolates), to isoleucine (11.5% of the isolates), or to valine (3.5% of the isolates). The introduction of the erg27(HydR3(+)) allele into a fenhexamid-sensitive strain by means of a replicative plasmid conferred fenhexamid resistance on the resulting transformants, showing that the mutations at position 412 are responsible for fenhexamid resistance. Weakly to moderately resistant isolates, termed HydR3(-), showed different point mutations between the strains in the sequenced regions of the erg27 gene, corresponding to amino acid changes between positions 195 and 400 of the protein. The erg27(HydR3(-)) alleles on the replicative vector introduced into a sensitive strain did not confer resistance to fenhexamid. Genetic crosses between HydR3(-) and sensitive strains showed strict correlation between the sequenced mutation in the erg27 gene and the resistance phenotypes, suggesting that these mutations are linked to fenhexamid resistance. The HydR3 mutations possibly modify the affinity of the 3-ketoreductase enzyme for its specific inhibitor, fenhexamid.
authors.affiliation: INRA UMR1290 BIOGER CPP, F-78000 Versailles, France. sabine.fillinger@versailles.inra.fr
authors.forename: Sabine
authors.initials: S
authors.lastname: Fillinger
authors.forename: Pierre
authors.initials: P
authors.lastname: Leroux
authors.forename: Christiane
authors.initials: C
authors.lastname: Auclair
authors.forename: Christian
authors.initials: C
authors.lastname: Barreau
authors.forename: Charbel
authors.initials: C
authors.lastname: Al Hajj
authors.forename: Danièle
authors.initials: D
authors.lastname: Debieu
mesh_terms: D000577:Amides / Q000494:pharmacology*; D019943:Amino Acid Substitution; D001483:Base Sequence; D020171:Botrytis / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D004271:DNA, Fungal / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics*; D004875:Ergosterol / Q000096:biosynthesis; D005602:France; D005659:Fungicides, Industrial / Q000494:pharmacology*; D005800:Genes, Fungal; D005858:Germany; D009154:Mutation; D010935:Plant Diseases / Q000382:microbiology; D027843:Vitis / Q000382:microbiology
chemical_list: D000577:Amides; D004271:DNA, Fungal; D005659:Fungicides, Industrial; C451426:N-(2,3-dichloro-4-hydroxyphenyl)-1-methylcyclohexanecarboxamide; D004875:Ergosterol
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