The Research On The Active Ingredients From Four Endophytic Fungi Based On Culture Strategy

Endophytes distributed in various tissue of plants.Endophytes colonized more highly in the stems of plant than in seeds,roots,leaves and flowers.The relationship between the endophytes and their host plant is favourable.Some active comounds from host plant could be found in its endophytes,which showed antibacterial,anti-inflammatory,anti-tumor and anti-insect activities.In this paper,the secondary metabolites from the endophytes of Tricholoma matsutake,Boletus edulis and Dendrobium were studied in order to find novel and active compounds,which could provide a new direction and theoretical basis for the development of new drugs.In the first chapter the progress on biological active metabolites from the endophytes of plant was summarized in recent years.In the second chapter the target strains were screened.40 strains were isolated from Tricholoma matsutake and 30 strains were isolated from Boletus edulis from Zhongdian Yunnan.Trichoderma harzianum SM-6 from Tricholoma matsutake and Phoma herbarum NM-46×1 from Boletus edulis were selected in 70 strains by chemical diversities.Nigrospora oryzae TPZ 10-1 from Dendrobium officinale and Irpex lacteus ZPY 45-2 from Dendrobium devoninum were screened for the conditions of culture.The coculture Nigrospora oryzae TPZ 10-1 with Irpex lacteus ZPY 45-2 was selected.In the third chapter the active metabolites from Trichoderma harzianum SM-6fermented by solid garlic were studied.Five known compounds were isolated and identified as:cis-octadec-9-enoic acid（1）,pyrrole-2-carboxylic acid（2）,2-（4-hydroxyphenyl）ethanol（3）,thymidine（4）,dibu-tylphthalate（5）.The antifungal and antifeedant activities were tested.Compounds 2 and 4 showed antibacterial activity against Aspergillus fumigatus with the MIC_S values 4μg/m L and 64μg/m L.Compounds2 and 3 showed antifeedant activity against silkworm larvae with antifeedant rates of 68%and 37%.The weight of silkworm larvae decreased by weight loss rate 5%,4%,and 2%after they were fed by compounds 2,3,and 4.（The weight loss rate of the blank control was 8%）In the fourth chapter the metabolites from the cocultures of Irpex lacteus ZPY 45-2and Nigrospora oryzae TPZ 10-1 were studied.3 compounds were isolated and identified as:nigrolactin（1*）,stigmast-4-ene-3-one（2）,（+）-（3S,6R,7R）-tremulene-6,11,12-triol（3）.Compounds 1*was a new compound.Compounds 1*and 2 have good antibacterial activity against Aspergillus fumigatus with MIC_Svalues 1μg/m L both.Compounds 1*and2 have weak antifeedant activity against silkworm larvae with antifeedant rates 9%and4%.The weight loss rate of silkworm larvae fed with compound 1*reached 5%,and the weight loss rate of the blank control was 8%.8 known compounds（4-11）from the cocultures of Irpex lacteus ZPY 45-2 and Nigrospora oryzae TPZ 10-1 were tested for the antifungal and antifeedant activities.compound 4,6,9-11 showed good antibacterial activity against Aspergillus fumigatus with the MIC_S value is 1-32μg/m L.Compounds 5and 10 showed good antifeedant activity against silkworm larvae,with antifeedant rates of 77%and 49%.The weight loss rate of the silkworm larvae fed with compounds 5,7,10 and 11 was between 4-9%.The crude extract from Irpex lacteus ZPY 45-2 and Nigrospora oryzae TPZ 10-1 cultured under different conditions were tested for the antifungal and antifeedant activities.The crude extract from TPZ 10-1/ZPY 45-2（potato:Dendrobium officinalis=2:1）has good antibacterial activity against Aspergillus fumigatus,with the MIC_S2μg/m L.The crude extract from TPZ 10-1（PDB）shows good antifeedant activity against silkworm larvae with the antifeedant rate 80%.The agravity rate of the crude extract from ZPY 45-2（Dendrobium officinalis）and TPZ 10-1（potato:Dendrobium officinali 2:1）were 8%,10%and 14%.In the fifth chapter the metabolites of Phoma herbarum NM-46×1 were studied.3compounds including 1 new compound:phombolin（1*）and 2 known compounds:6-methylsalicylic acid（2）,3-hydroxybenzyl alcohol（3）were isolated and identified.Compounds 1*-3 show good antibacterial activity against Aspergillus fumigatus with the MIC_S 8μg/m L,1μg/m L and 64μg/m L.Compound 1*and 3 have good antifeedant activity against silkworm larvae with the antifeedant rate 88%and 57%.