Isolation of Tricholoma matsutake and T. bakamatsutake cultures from field-collected ectomycorrhizas

Tricholoma matsutake was isolated into pure cultures from field samples of ectomycorrhizas on Pinus densiflora. The mycorrhizal tips were collected at different times of the year from a colony of T. matsutake in a P. densiflora stand, The mycorrhizal tips were continuously washed with sterilized distilled water and diluted Tween 80 solution, surface-sterilized with calcium hypochlorite solution, and inoculated on several kinds of nutrient agar media, Most of the mycorrhizal tips collected in winter and spring produced colonies that were morphologically similar to cultures of T. matsutake isolated from basidiocarps, The identity of isolates obtained from mycorrhizas was further confirmed to be T. matsutake based on fungal morphology and RFLP patterns of PCR amplified rDNA, The feasibility of T. bakamatsutake isolation into pure culture from ectomycorrhizas on Quercus serrata was also confirmed. These results indicated that mycelium of matsutake mushrooms can be isolated into pure culture from ectomycorrhizas at different times of the year, Mycorrhizas of both T. matsutake and T. bakamatsutake were not observed to have any specific association with soil fungi such as Mortierella spp