Detection and species discrimination using rDNA T-RFLP for identification of wood decay fungi

In the present work PCR technology was used as a tool to detect the early stages of wood decay and was compared with microscopic evaluation. The wood decay fungi Postia placenta and Coniophora puteana were detectable in interior wood samples by terminal restriction fragment length polymorphism (T-RFLP) after 2 weeks of incubation with monocultures, while microscopic detection of hyphae was not possible until after 7 weeks. A potential problem when fungal communities are studied with T-RFLPs of rDNA is that intra-specific variation complicates data analysis. In this work, we show that intra-specific sequence variation in the internal transcribed spacer of the rDNA in Coniophora puteana allows T-RFLP identification of this species. This is due to intra-specific variations in fragment length, in combination with the absence of point mutations in the selected restriction sites.