PCR cloning and detection of point mutations in the eburicol 14alpha-demethylase (CYP51) gene from Erysiphe graminis f. sp. hordei, a "recalcitrant" fungus.
Molecular studies of some micro-organisms are hampered by the difficulty of obtaining sufficient amounts of nucleic acids. A cloning strategy based on PCR has therefore been used to clone the eburicol 14alpha-demethylase (CYP51) gene of the obligate fungus Erysiphe graminis f. sp. hordei (Egh) using minute amounts of genomic DNA. The CYP51 gene encodes the enzymatic target of a major group of fungicides. Sequencing CYP51 from different Egh isolates revealed the occurrence of two alleles for this gene. An allele-specific PCR assay was developed to detect each CYP51 allele.
Délye
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Unité de Recherches Intégrées sur la Vigne, Institut National de la Recherche Agronomique, Domaine de la Grande Ferrade, B. P. 81, F-33883 Villenave d'Ornon cédex, France. delye@bordeaux.inra.fr
Bousset
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Corio-Costet
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D000483:Alleles; D000595:Amino Acid Sequence; D001203:Ascomycota / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005784:Gene Amplification; D005800:Genes, Fungal / Q000235:genetics*; D014644:Genetic Variation; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D058886:Sterol 14-Demethylase
D018076:DNA, Complementary; D004271:DNA, Fungal; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D058886:Sterol 14-Demethylase