Development of PCR method for fast detection of Ophiostoma floccosum in wood chips

Ophiostoma floccosum is one of blue stain fungi that cause cosmetic damage in softwood. To develop a molecular maker for the detection of O. floccosum, we designed Oflo1 and Oflo2 primers based on the β-tubulin gene. PCR assay with eleven isolates ofO. floccosum and nine other blue stain and discoloring fungal species (moulds) proved that the designed primers specifically amplified a 333 bp sized DNA band only from genomic DNA of O. floccosum isolates. Together with microwave heating extraction of spore DNA, we could detect O. floccosum within 2 h on artificially inoculated wood chip. We believed that the developed PCR method will be useful where detection of target species such as O. floccosum is required.   Key words: Ophiostoma floccosum, Species-specific primer, β-tubulin gene.