Studies On Isolution,purification And Its Secondary Metabolites Bioactivity Of Endophytic Fungi From D.Tangutica,S.hexandrum And E.brevicornu

Endophytic fungi can produce the same or similar compounds with their host during long-term cooperative coevolution and widely distributed in plants,whose secondary metabolites of endophytic fungi can be thought of as a broad application prospect and potential of the compound library in medicine,agriculture and other related fields.In this case,Daphne tangutica Maxi.,Sinopodophyllum hexandrum(Royle)Ying and Epimedium brevicornum Maxim.were chosed to be research object.The resources of endophytic fungi from Daphne tangutica Maxi.,Sinopodophyllum hexandrum(Royle)Ying and Epimedium brevicornum Maxim.and their bioactives were investigated.The results were as follows:1.Isolation medium was PDA and cultured at 28? in the incubator,endophytes obtained were 30 strains from fresh Daphne tangutica Maxi.,Sinopodophyllum hexandrum(Royle)Ying and Epimedium brevicornum Maxim.These endophytic fungi have various colony morphology.Through observing their morphology,9 strains produced spores were preliminary identified as Fungi Imperfecti,and 2 strains are to Alternaria,the other 7 strains are to Fusoma,Didymopsis,Piricularia,Cephalosporium,Moniliara,Aspergillus and Fusarium.2.A total of 90 samples of ethyl acetate,normal butanol and methanol from three plant species were obtained and all the 90 samples of antibacterial,antioxidant,and?-glycosidic enzyme inhibitory activity were determined.The activities of antibiosis were tested by filter-paper method(All the samples' concentration are 10 mg/m L).The result shows that the ethyl acetate extraction from strain RX-S-102 has a very strong antibiosis to Staphyloccocus aureus Rosenbach,Bacillus licheniformis,Escherichia coli,Pseudomonas aeruginosa and Klebsiella pneumoniae,whose diameter of the zone of inhibition towards these 5 kinds of indicator bacteria are at the range of 22.3~28.0mm and the MIC towards these 5 kinds of indicator bacteria are at the range of 0.025~0.100 mg/m L.Meanwhile,the ethyl acetate extraction from strain RX-S-105 has a strong antibiosis to Escherichia coli and Klebsiella pneumoniae,the n-butyl alcohol extraction from strain GJ-S-208 has a strong antibiosis to Staphyloccocus aureus Rosenbach and the methyl alcohol extraction from strain GJ-S-203 has a strong antibiosis to Bacillus licheniformis,the n-butyl alcohol extraction from strain YYH-S-301 has a strong antibiosis to Escherichia coli In addition,in total 90 samples,in addition to the n-butyl alcoholextraction from strain RX-S-106 did not show obvious inhibitory activity to all 5 kinds of indicator bacteria,the rest of the 89 samples at least one of the five kinds of indicator bacteria showed antimicrobial activity.Antioxidation experiments were tested by the potassium ferricyanide reduction method,the salicylic acid method and so on(All samples' concentration are 0.1 mg/m L).The result shows that there are 6 extracted samples' VCEAC is over 300 ?g/mg.The highest data is the ethyl acetate extraction from strain RX-S-104,GJ-S-203 and YYH-S-304 and their VCEAC are 973.39 ?g/mg,735.08 ?g/mg and 710.39 ?g/mg.The EC50 of Vc and all these 3 simples are 0.1164 mg/m L,0.1266 mg/m L,0.1732mg/m L,0.1796 mg/m L.12 extractions DPPH clearance rates reach 80%.The DPPH clearance rates of the methyl alcohol extraction from strain GJ-S-208 is 92.79% and the DPPH clearance rates of the ethyl acetate extraction from strain GJ-S-208 is90.26%.The IC50 of Vc and all these 2 simples are 0.0054 mg/m L,0.0541 mg/m L and0.0563 mg/m L.All samples show low hydroxyl free radical activities.The ?-Glycosidase activity was assayed by the improved PNPG method(All the samples' concentration are 1 mg/m L).The result shows that all the 90 samples have?-glucosidase inhibitions.There are 35 samples which have much than 80% inhibitions and 17 samples which have much than 80% inhibitions towards ?-Glycosidase.3.The fermentation products chemical constituents of RX-S-102 were isolated.In the end,2 compounds were isolated from the ethyl acetate fraction,which was identified to ?-sitosterol and umbelliferone.The results of this study would offer the foundation to the further development and utilization of endophytic fungi Daphne tangutica Maxi.,Sinopodophyllum hexandrum(Royle)Ying and Epimedium brevicornum Maxim.three kinds of medicinal plants.