Development of a multiplex allele-specific primer PCR assay for simultaneous detection of Qol and CAA fungicide resistance alleles in Plasmopara viticola populations

摘要

BACKGROUND: DNA-based diagnosis has become a common tool for the evaluation of fungicide resistance in obligate phytopathogenic fungus Plasmopara viticola. RESULTS: A multiplex allele-specific primer PCR assay has been developed for the rapid detection of fungicide resistance in P. viticola populations. With this assay, a glycine-to-alanine substitution at codon 143 of the P. viticola cytochrome b gene, which conferred Qol fungicide resistance, and a giycine-to-serine substitution at codon 1105 of the P. viticola cellulose synthase gene PvCesA3, which conferred CAA fungicide resistance, were detected simultaneously. CONCLUSION: it is suggested that the present assay is a reliable tool for the rapid and simultaneous detection of Qol and CAA fungicide resistance alleles in P. viticola populations. The assay required only 2 h from the sampling of symptoms to the detection of resistance alleles to both fungicides.