An Allele-specific PCR Assay for Detecting Azoxystrobin-resistant Alternaria Isolates from Pistachio in California

Alternaria late blight caused by Alternaria spp. in the alternata, tenuissima and arborescens species‐groups is one of the most common fungal diseases of pistachio in California. A single point mutation resulting in the replacement of a glycine by an alanine at codon 143 (G143A) in the mitochondrial cytochrome b (cyt b) has been found in all azoxystrobin‐resistant isolates of these three species from California pistachio. In this study, a pair of allele‐specific polymerase chain reaction (PCR) primers was developed to detect this point mutation. The allele‐specific PCR assay coupled with a rapid DNA extraction method could detect azoxystrobin‐resistant Alternaria isolates in a few hours. The allele‐specific PCR method was also reliable for detecting azoxystrobin‐resistant Alternaria directly in both laboratory‐inoculated and naturally infected pistachio leaves.