Cloning and sequence analysis of the eburicol 14alpha-demethylase encoding gene (CYP51 ) from the Japanese pear scab fungus Venturia nashicola

The slow growth of many lamentous fungi in vitro prevents rapid isolation of large quantities of genomic DNA, thereby hampering molecular studies. Using a polymerase chain reaction (PCR)-based adapter ligation cloning technique that requires only nanogram quantities of genomic DNA we have successfully cloned the complete CYP51 gene encoding the eburicol 14 a demethylase, the target for DMI (Demethylation-inhibiting) fungicides, from the Japanese pear scab fungus, Venturia nashicola. Analysis of predicted amino acid sequences of CYP51s from strains less sensitive to DMIs revealed no alterations when compared to the sensitive reference strain.