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目的:探讨银杏内生真菌的分离方法、菌种鉴定及其代谢产物粗提物的体外抗真菌、抗氧化活性,为进一步深入研究银杏内生真菌及其代谢产物的活性提供依据。方法:首次对天目山地区野生银杏植株树皮、枝条、树叶进行了内生真菌的分离;采用微量稀释法测试银杏内生真菌粗提物抗病原真菌活性。采用DPPH法测试银杏内生真菌粗提物抗氧化活性。同时采用分子生物学方法对筛选出的活性内生真菌进行鉴定。结果:研究发现,共计9株(16.1%)内生真菌粗提物的抗白色念珠菌Candida albicans、新生隐球菌Cryptococcus neoformans、红色毛癣菌Trichophyton rubrum、薰烟曲霉菌Aspergillus fumigatus活性较好。其中以天目山野生银杏树皮中分离得到的青霉属T-1-2-1菌株粗提物抗真菌活性最佳,对4种供试真菌均具有一定的抗菌活性,其抗红色毛癣菌的MIC80为4 mg.L-1,与阳性对照氟康唑的抗菌活性相当;共计5株(8.9%)内生真菌的抗氧化活性较好,其中,以毛壳属T-3-2-2、炭角菌属T-6-5-7菌株抗氧化活性最好。结论:银杏内生真菌是一个潜在的、丰富的抗菌、抗氧化新药资源库,值得进一步研究和开发。
OBJECTIVE: To study the isolation method of endophytic fungi, the identification of strains and the antifungal and antioxidant activities of the crude extracts of their metabolites in order to provide basis for further study on the activity of endophytic fungi and their metabolites. Methods: Endophytic fungi were isolated from the bark, shoots and leaves of wild ginkgo trees in Tianmu Mountain area for the first time. The antifungal activity of the crude extracts of endophytic fungi from Ginkgo biloba was tested by micro-dilution method. Antioxidant Activity of Crude Extract from Endophytic Ginkgo Biloba by DPPH Method. At the same time, molecular biological methods were used to identify the active endophytic fungi. Results: A total of 9 (16.1%) crude extracts of endophytic fungi Candida albicans, Cryptococcus neoformans, Trichophyton rubrum and Aspergillus fumigatus were found. Among them, the crude extract of Penicillium T-1-2-1 isolated from the wild ginkgo bark of Tianmu Mountain had the best antifungal activity, and all of the tested fungi had antimicrobial activity. The anti-Trichophyton rubrum The MIC80 was 4 mg.L-1, which was comparable to that of the positive control fluconazole. A total of 5 strains (8.9%) of endophytic fungi had better anti-oxidative activities, of which the genus Tritillaria genus T-3-2- 2, carbon antagonism T-6-5-7 strain the best antioxidant activity. Conclusion: Ginkgo biloba endophytic fungus is a potential and abundant resource of anti-bacterial and anti-oxidant drugs, which deserves further research and development.