Heterologous expression of the P450 sterol 14α-demethylase gene from Monilinia fructicola reduces sensitivity to some but not all DMI fungicides

The cytochrome P450 sterol 14α-demethylase gene (MfCYP51) from Monilinia fructicola (G. Wint.) Honey was cloned and sequenced. The gene was 1680 bp in length (including introns) and was predicted to have two introns of 54 and 57 bp. The nucleotide sequence was 82.1, 53.4, 47.1, 45.1, and 33.6% and the amino acid sequence was 89.7, 76.1, 76.1, 71.8, and 66.9% identical to the CYP51 genes from Botrytis cinerea, Tapesia yallundae, T. acuformis, Erysiphe graminis, and Uncinula necator, respectively. Expression of MfCYP51 in PDR5::TN5 deficient Saccharomyces cerevisiae resulted in reduced sensitivity of the yeast transformants to myclobutanil but not to propiconazole, fenbuconazole or tebuconazole. A wildtype population of 33 M. fructicola isolates was significantly less sensitive to myclobutanil than to propiconazole, fenbuconazole, and tebuconazole. The sensitivity of the isolates to myclobutanil and the three other DMI fungicides included in this study was correlated positively, suggesting a similar or identical mode of action. The low sensitivity in M. fructicola wildtype isolates to myclobutanil could result from a less effective binding potential of the fungicide to the 14α-demethylase.