STUDIES ON THE ISOLATION, CULTURE AND DNA IDENTIFICATION OF MYCELIA OF TRICHOLOMA MATSUTAKE
松口蘑菌丝体的分离和RAPD-PCR分析

The tissue isolation for Tricholom matsutake(S Ito et Imai) Sing were made with 8 media in 810 test tubes from different positions of 9 basidiocarps of different source and from mycorrhizae, and soil with the fungi in the studies. The results showed that 94 test tubes of slow-growing mycelia were isolated from lamellae and their success percentages of isolation with media PDAS, PDAW, BM, PDA were 74.4%, 355%, 15.6% and 8.9% respectively. The fast-growing mycelia were easily got from the mycorrhizae and soil related to matsutake. The isolates with different culture characteristics were appraised through DNA fingerprinting comparison with matsutake basidiocarps collected from Jilin province, China and reference isolates presented by matsutake research workers of China and Japan, in which RAPD (Random Amplified Polymorphic DNA)-PCR patterns were sharply prepared using 17 arbitrary decamer nucleotide primers screened. The statistical data indicated that all slow-growing mycelia isolated from lamellae had the same DNA fingerprinting patterns as their origin basidiocarps tissues such as pileus (containing lamellae) and stipe, whose similarity coeffecients all were 1.000, and were therefore identified as true Tricholoma matsutake. However, the fast-growing mycelia or yeast colony were identified as not matsutake. The results suggested that matsutake and its own mycelia have DNA homogeneity, and there exists no any other microbe in the basidiocarps. The results also demonstrated that all matsutake from east China and reference isolates of matsutake from southwest China and Japan were one same species Tricholoma matsutake, whose DNA similarity coeffecients varied from 0.934 to 0.994.