Function of the genetic element "Mona' associated with fungicide resistance in Monilinia fructicola

The genetic element Mona' has been shown previously to be associated with resistance to demethylation inhibitors (DMIs) in Monilinia fructicola. In this study, the promoter activity of the Mona' element was demonstrated genetically and the activity was narrowed down to a 20-bp active region through a series of deletions. Mona' knockout transformants (Mona) were generated from DMI-resistant isolate Bmpc7, and EC50 values and expression of the MfCYP51 gene were found to be reduced in transformants compared with the parental isolate. When the Mona' element was inserted into the upstream region of the MfCYP51 gene of the DMI-sensitive isolate HG3, the EC50 values and expression of the MfCYP51 gene increased in the transformants compared with the parental sensitive isolate. These results indicate that the Mona' element determines the DMI fungicide resistance through the up-regulation of the expression of the downstream MfCYP51 gene. No fitness penalty was observed in knockout and insertion transformants, i.e. transformants showed similar mycelial growth rate, sporulation and ability to cause lesions on fruit compared with their parental isolates, suggesting that the Mona' element does not affect basal life activities.