NOVELTY - Separating Tricholoma matsutake comprises e.g. choosing Tricholoma matsutake fruiting body, scrubbing the fruiting body, placing in sterilized laminar flow hood, breaking the fruiting body, tearing the skin surface of the cap, sterilizing, grabbing the edge of the cap, gripping tissue blocks, dipping in small amount of non-solidified plate culture medium, sticking in a middle surface of the petri dish cover, culturing and spraying the mature spores on to the medium for germination into pure mycelium.    USE - The method is useful for Separating Tricholoma matsutake (claimed).    ADVANTAGE - The method: has success rate of 90% and high germination rate; is simple and environmentally friendly; and requires less raw materials.    DETAILED DESCRIPTION - Separating Tricholoma matsutake comprises choosing Tricholoma matsutake fruiting body, where fruiting body of typical appearance, medium size, fertile thick fungal meat, normal colored and not well opened fruiting body is choosed, scrubbing the fruiting body 2-3 times with 75% alcohol preps, placing in sterilized laminar flow hood, breaking the fruiting body at the junction of cap and stipe, tearing the skin surface of the cap, sterilizing the exposed portion of fungal meat and gills, grabbing the edge of the cap with sterile tweezers, gripping 1-2 cm3 size tissue blocks, where the gripping tissue block comprises both upper fungal meat part and lower gill part, dipping the tissue blocks into small amount of non-solidified plate culture medium at the temperature of 60 degrees C such that the gill portion is not in contact with the medium, sticking the tissue blocks in a middle surface of the petri dish cover where the gill part is down and the distance between the tissue blocks and medium is 0.6 cm, covering with sealing lid, culturing upright in dark at the temperature of 25 degrees C, where the meat part of the tissue block strain supply nutrients to the gills, gills mature and spray spores, and spraying the mature spores on to the medium for germination into pure mycelium.