PCR cloning and detection of point mutations in the eburicol 14α-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 14 alpha-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.