Studies On The Isolation, Culture And DNA Identification Of Mycelia Of Saprophytic And Symbiotic Edible Fungi

Spawn is necessary for both cultivation of edible fungi and tameness of wild mushroom. The reliable method of spawn identification is to produce carpophores by the culture, which is not only time-consuming for domestic fungi but also unuseful for untamed wild mushroom. The DNA identification method was developed for mycelial isolates of saprophytic and symbiotic edible fungi in the studies by comparing the DNA fingerprintings of isolates with their original carpophores.l.DNA identification of isolates of Lentinula mushroom. The DNA fingerprinting patterns were made by RAPD-PCR using 12 random primers. The results showed that there are same RAPD fingerprintings among the mycelial isolates, pileus (including lamellae), lamellae, and stipe from one basidiocarps of Lentinula mushroom, also between the mycelial spawn and its culturing basidiocarps. The results also suggested that there are a little DNA variation among different origin basidiocarps, and their similarity coefficients vary from 0.886 to 0.986.2.DNA identification of isolates of Pleurotus mushroom. The RAPD-PCR analyses were conducted with different colour basidiocarps from different origin Pleurotus mushrooms. The results showed that there are same DNA fingerpringing patterns among the mycelial isolates, pileus (including lamellae)and stipe from one basidiocarps, but the similarity coefficients between different origin and colour basidiocarps vary from 0.779 to 0.976.3.Survey of vegetation of mountain growing Tricholoma matsutake. The vegetation survey were conducted in a mountain located hi Longjng , Jilin province. 74 species belonging to 38 family plants, and 15 species belonging to 8 family large fungi were identified and recorded. Those easily found plants and fungi are Pinus densiflora, Quercesmongolica, Lespebeza bicolor, Rhododendrum chrysanthum, Melampyrum roseum, Ramaria stricta, Cortinarius collinitus, etc, among which, 22 species of plants and 3 species of mushrooms were first reported occurring beside fairy rings of matsutake.4.Studies on DNA polymorphisms of symbiotic edible fungi, Tricholoma matsutake. The DNA fingerprinting of wild baidiocarps of matsutake, collected from the scenes of major production regions in China, were analyzed based on RAPD (Random Amplified Polymorphic DNA)-PCR patterns were optimized by example experiments of two DNA templates in this study. The RAPD-PCR conditions were optimized by example experiments of two DNA templates from different ecological regions, and 18 of 40 arbitrary decamer nucleotide primers were also screened with good amplication and repetition. The 17 sets of DNA fingerprinting patterns were sharply made for 20 basidiocarps tested. The similarly coeffecients of DNA fingerprintings among the materials tested ranged from 0.50 to 0.90. And 15 of 20 pine mushrooms tested were clustered together at 0.72 similarity level by UPGMA cluster analysis, and were demonstrated to be the same species Tricholoma matsutake, distrubuted in Yunnan, Sichuan and Jilin provinces of China. The DNA fingerprinting patterns in this study are made to help identification of wild matsutake resources and their tissue isolates under various ecological environments fit for the artificial taming of matsutake.5. Mycelia isolation and culture of Trichotoma matsutake. Tissue isolation of different positions of 9 matsutake were made with 8 media in 810 test tubes. The results showed that lamellae were fit tissue for mycelia isolation, and success percentages of mycelia isolation with media PDAS, PDAW, BM, PDA are 74.4%,35.5%,15.6% and 8.9% respectively. The results also showed that stipe, mycorrhizae, and soil with the fungi are not fit for Tricholoma matsutake isolation.6. DNA identification of Tricholoma matsutake isolates. 24 samples including matsutake basidiocarps and different origin isolates were analyzed with RAPD fingerprinting comparison using 17 arbitrary decamer nucleotide primers. The results showed that all slow-growing mycelia isolated from lamellae have the same DNA fingerprinting...