Detection of the ectomycorrhizal fungus Tricholoma matsutake and some related species with specific ITS primers

Abstract

Oligonucleotide primers (Tm 1 and Tm4) were designed to amplify a 447–448 base pair fragment, comprising sections of the rDNA internal transcribed spacers (ITS) and the entire 5.8S rDNA, of Tricholoma matsutake. PCR products of predicted size were produced for six of eight isolates of T. matsutake from across its natural range in Asia, and for isolates of some closely related fungi including T. bakamatsutake, T. magnivelare, and T. caligatum. The closely related T. robustum could be discriminated from L. matsutake by PCR fragment size. No PCR products were produced where the primers were tested against 16 species of ectomycorrhizal fungi associated with Pinus spp. in the Southern Hemisphere. The specific primers were also used successfully to produce PCR products from matsutake infected roots collected in natural forests in China and Japan, and from pure culture synthesised Pinus radiata-T. matsutake material. These primers will be useful in research directed at establishing matsutake in the Southern Hemisphere, and also have the potential to be applied to the study of matsutake within its natural range.