| Objective: To study the species and distribution of endophytic fungi of Bozhou Paeonia lactiflora Pall in different tissues and different seasons,and to analyze the dominant bacteria and select the anti-pathogenic microorganism dominant bacteria.Further analysis the metabolites,in order to separate lead compounds own novel structure and excellent biological activity.Our findings are beneficial for laying a foundation for the further research and quality control of the Dao-di herbs resources of Bozhou P.lactiflora Pall.Methods: The endophytic fungi of Bozhou P.lactiflora Pall were isolated and identified from different tissue parts in different seasons by using tissue block separation and molecular biology.The species diversity and community structure of endophytic fungi were analyzed by biological statistics.The crossing method had been used to determine the inhibitory effects of dominant strains against plant pathogenic fungi.The solid fermentation technology was used to ferment dominant strains,separation and identification of solid fermentation products were used by chromatographic methods and modern spectroscopic techniques.Results: A total of 509 endophytic fungi strains were isolated from Bozhou P.lactiflora Pall,which belonged to 18 genus and 31 species.The species of endophytic fungi were more abundant in summer and autumn than other seasons,indicated from isolation frequency,richness index and diversity index.Meanwhile,the most abundant endogenous fungi were isolated from root tissues.Among them,Penicillium,Alternaria and Fusarium strains had high isolation frequency,A l t e r n a r i a a l t e r n a t a,C e r a t o b a s i d i u m r a m i c o l a,A s p e r g i l l u s s p.,Talaromyces minioluteus,Acrocalymma aquatica,Cladasporium uredinicola,F u s a r i u m e q u i s e t i,Colletotrichum spaethianum,Fungi endophyte and P e n i c i l l i u m m a r i a e-c r u c i s,these ten fungi were the dominant genus with more than 2% separation frequency.The antibacterial activity test showed that the metabolites from all these endophytic fungi possess widespread anti-phytopathogenic fungi activity.Among them,the ethyl acetate extract of the metabolites of P.mariae-crucis had obvious antibacterial effect on six pathogenic fungi,and the MIC to Staphylococcus aureus is 64 μg/m L.The antibacterial rate of A.alternata metabolites against Pepidoides spp.reached 75%.The metabolites of A.sp.,T.minioluteus,A.aquatica and F.equiseti had temperate antibacterial effects against 6 pathogenic fungi,which basically reached 25%.The MIC of ethyl acetate extract of Ascochyta phacae metabolites strain on Candida albicans was128 μg/m L.A total of 45 compounds were isolated and identified from the two dominant strains,as: alternate A(1),alternate B(2),alternate C(3),alternate D(4),alternate E(5),alternate F(6),TCA ma1(7),TCA ma2(8),TCA ma3(9),TCA ma4(10),TCA ma5(11),TCA3 b(12),TCA3 a(13),TCA A(14),methy 4-acetamido-3-hydroxybenzoate(15),(E)-methyl-5-hydroxy-3-methylpent-2-eno(16),isobenzofuranone A(17),4-hydroxyacetophenone(18),6-hydroxy-isosclerone(19),talarojlavone(20),7-hydroxy-2-hydroxymethyl-5-methyl-4H-chromen-4-one(21),alternarienonic acid(22),2,5-dimethyl-7-hydroxychromone(23),3-hydroxy-5-methyoxybenzoic acid(24),5-hydroxyepialtenuene(25),alternariol(26),methyl m-hydroxybenzoate(27),stemphyperylenol(28),methyl-2-(4-hydroxyphenyl)propanoate(29),altenusin(30),altenuene(31),djalonensone(32),3-hydroxyalternariol 5-O-methyl ether(33),altenuisol(34),1-deoxyrubralactone(35),penicmariae-crucis A(36),penicmariae-crucis B(37),penicmariae-crucis C(38),penicmariae-crucis D(39),(E)-2-methylhex-3-enedioic acid(40),(22E)-5a,8a-epidioxyergosta-6,22-dien-3b-ol(41),β-Sitosterol(42),O-methylcurvulinic acid(43),methyl,2-(2-acetyl-3-hydroxyl-5-methoxyphenyl)acetate(44),methyl 2-acetyl-3,5-dihydroxyphenylacetate(45).Compounds 1-35 were isolated from metabolites of the endophytic fungi Alternaria alternat and compounds 1-11 were new compounds.Compounds 36-45 were isolated from the metabolites of the endophytic fungi Penicillium mariae-cruci and Compounds 36-39 were new compounds,compound 40 was a new natural product.Compound 36-45 was isolated from the metabolites of the endophytic fungi P.mariae-crucis first time.Conclusion:There are abundant endophytic fungi in Bozhou P.lactiflora Pall.and there are some differences between different seasons and different tissues in different tissues.Secondary metabolites of different endophytic fungi have different antibacterial activities.Metabolites are abundant and diverse from two Endophytic Fungi A.alternata and P.mariae-crucis. |
PDF Full Text Request