Abstract: Four strains of endophytic fungi were isolated and purified from wild bodies of Leccinum crocipodium, Boletus speciosus, and Boletus bicolor by tissue separation method. Microscopic observation of its colonies, hyphae, and conidia morphology, and molecular biological identification of rDNA ITS sequence results showed that: Strain LCTG-12 was Trichoderma gamsii, LCCK-6 was Candida kruisii, BSHC-18 was Hypomyces chrysospermus, BBFO-10 was Fusarium oxysprum. The biological characteristics of four endophytic fungi studied, among them, the mycelium of Trichoderma gamsii growed faster, cultured on CPDA medium at 25 ℃ for 5 days, mycelium could cover the entire medium. In liquid culture, the strain could obtain 4.13 g/L of dry biomass of mycelium at 25 °C, shaker speed 160 r/min, pH5.5, and 8 days. The results of the flat panel stand-off experiment: LCTG-12 Trichoderma gamsii had an antagonistic effect on the growth of BSHC-18 Hypomyces chrysospermus and BBFO-10 Fusarium oxysprum, at the same time, BSHC-18 Hypomyces chrysospermus and BBFO-10 Fusarium oxysporum had an antagonistic effect. Therefore, the endophytic fungi Trichoderma gamsii of boletus had an antagonistic effect on spoilage fungi and may be an excellent strain for biological control.