A molecular diagnostic assay for the detection and identification of wood decay fungi of conifers

Summary Ten taxon‐specific primers were designed to amplify the Internal Transcribed Spacer of the r RNA operon of several important decay fungi of coniferous wood, including A rmillaria spp., E chinodontium spp., F omitopsis pinicola , F uscoporia torulosa , H eterobasidion annosum sensu lato ( s.l .), O nnia spp., P haeolus schweinitzii , P hellinus weirii s.l ., P holiota spp. and P orodaedalea spp. Primers designed in this study and in a previous one for the identification of L aetiporus sulphureus and S tereum spp. were combined in two multiplex PCR s, which were tested for efficiency and specificity, and detected at least 1 pg of fungal target DNA . Target DNA at concentrations of 10 −1  pg or lower can be detected with this assay using SYBR ® G reen R eal‐ T ime PCR . Validation assays performed on 129 naturally infected wood samples or fruiting bodies confirmed the reliability of the multiplex PCR ‐based diagnostic method. This method represents a simple and rapid diagnostic tool for the detection of a number of destructive wood decay fungi of conifer wood.