Isolation,Identification And Their Bioactivity Of Endophytic Fungi From Five Plants

Endophytic fungi could not only produce active components which were same as those of plants?such as anti-cancer,anti-bacterial,etc.?,but also produce novel structural compounds,which became an important source of new bioactive natural products.In order to screen biologically active substances,we focused on the isolation of Endophytic Fungi from five plants and their bio-activities,including anti-MAO,anti-ACh E,insecticidal and anti-bacterial activities.The research contents and results are as follows:A total of 48 strains were isolated from five plants by tissue-culture method.Among them,8 strains were isolated from Berberidis poiretii,6 strains were isolated from Lamium amplexicaule,8 strains were isolated from Euphorbia micractina,12strainswere isolated from Hypericum perforatum,14 strains were isolated from Lonicera tangutica.By fermentation and extraction,96 crude extracts were obtained,and their biological activity were evaluated.The results showed that BGC-1-L,BGC-2-L,BGC-5-L,LSRD-12-L,LSRD-1-L and XYXB-1-L showed monoamine oxidase inhibitory activity with IC₅₀0 of128.30,72.08,109.59,112.43,146.07 and 63.55?g/m L,respectively.When the final concentration was 100?g/m L,GYLQ-3-L has weak acetylcholinesterase inhibitory activity with inhibition rate of 41.19%.When concentration of crude extracts were 2 mg/m L,7 extracts showed remarkable insecticidal activity of Tetranychus cinnabarinus with inhibitory rate of more than85%at 48h,which were GQDJ-4-L,GYLQ-1-L,GYLQ-2-L,XYXB-3-L,BGC-2-L,XYXB-8-L,and BGC-4-S with death rates of 86.67,90.00,93.33,93.33,93.33,96.67and 93.33%,respectiviely.9 extracts showed remarkable insecticidal activity against Brevicorynre brassicae with inhibitory rate of more than 90%at 48 h,which were LSRD-1-L,LSRD-6-L,LSRD-6-S,LSRD-12-L,GQDJ-3-L,GQDJ-5-S,GYLQ-3-S,XYXB-5-S and XYXB-8-L with death rates of96.67,90.00,90.00,96.67,90.00,90.00,96.67,90.00 and 93.33%.The inhibitory activities against Phytophthora infestans and Phytophthora capsici of all endophytic fungi was determined by the method of toxic culture.When the concentration was 2 mg/m L,3 extracts showed inhibitory activity against Phytophthora capsici with inhibitory rate of more than60.00%,which were GYLQ-11-L,LSRD-8-S and SKZ-5-L with inhibitory rates of60.13,69.93 and 63.66%.2 extracts showed inhibitory activity against Phytophthora infestans with inhibitory rate of more than 55%,which were GQDJ-5-L and BGC-5-L with inhibitory rates of 55.92 and 55.26%.Twelve compounds were isolated andpurified from strain BGC-2 by chromatographic methods,including macroporous resin,MCI,silica gel column chromatography,TLC,HPLC,etc.The structures were identified as Cyclotryprostatin A?1?,Demethoxyfumitremorgin C?2?,Fumiquinazoline J?3?,Spirotryprostatin A?4?,Cyclotryprostatin B?5?,?-trans-2,5,15-trihydroxybergamot-10-ene?6?,Pesurotin G?7?,Cephalimysin A?8?,CS-E?9?Fumitremorgin,C?10?,Lupeol?11?and 1,2-benzenedicarboxylic acid,dibutyester?12?.Compound 7,9 showed relatively high anti-MAO activity with IC₅₀0 of 49.72±0.44,62.48±1.67?g/m L.Compound 4,8,10showed relatively high anti-acetylcholinesterase activity with IC₅₀0 of 17.91±0.02,33.49±1.23,24.37±0.77?g/m L.By combination morpholog with molecular biology methods,strains BGC-1?BGC-2?LSRD-1?LSRD-12 and XYXB-1 were identified as Alternaria sp.,BGC-3was identified as Penicillium sp.,and LSRD-5 was identified as Aspergillus sp.