Multiplex PCR assay for simultaneous detection of MBI-D and QoI resistance in rice blast fungus

For sustainable control of rice blast with fungicides, efficient monitoring of the emergence and spread of fungicide-resistant isolates is needed. We developed simple and reliable PCR-based DNA markers to detect isolates resistant to melanin biosynthesis inhibitor targeting scytalone dehydratase (MBI-D) and quinone outside inhibitor (Q_oI) fungicides. Through the use of DNA templates prepared from mycelia on filter paper or from infected leaves, these markers enable rapid (a few hours) genotyping of point mutations that confer resistance. The developed multiplex marker detected resistance to both MBI-D and Q_oI in a single PCR and further reduced the time needed for diagnosis.