Study On Secondary Metabolites Of Endophytic Fungus Strain Fusarium Solani A2 And Fimetariella Rabenhorstii A20 From Aquilariae Lignum Resinatum

Recent researches show that, Endophytic fungi,which lives within the healthy plant tissue, plays an important role in the plant microbial ecosystem and makes the host plants do not show obvious symptoms of infection. Endophytic fungi can produce the same or similar bioactive substances of their host. Endophytic fungi, they live in their host, which is beneficial to both the Endophytic fungi and its host. More and more coming researches indicate that those substances with obvious bioactive come from Endophytic fungi, that makes Endophytic fungi becomes the significant source to examine the new bioactive substances.Chinese eaglewood,which will produce some black resin when being hurt by external stimuli such as physics,chemistry or fungal, is the fragrant wood of Aquilaria sinensis(Lour).Gilg.In previous research, endophytic fungi strain Fusarium solani A2 and Fimetariella rabenhorstii A20 were isolated from Aquilaria sinensis. In the virto experiment of Aquilaria sinensis, the result shows that Fusarium solani A2 can induce aloes. And Fimetariella rabenhorstii A20 can produce substances like aloes.In order to explore the effect of the fungi on the Aquilaria sinensis,we establish an in vitro experimental method,chemical constituents of Aquilaria sinensis excised twig inoculated with Et OAc extracted from the Fusarium solani A2 and Fimetariella rabenhorstii A20 were analyzed by GC-MS.Some terpenoids were detected in experimental group.No cheemical constituent of agarwood in control group was detected.The results showed that Et OAc extracted from the Fusarium solani A2 and Fimetariella rabenhorstii A20 can induce Aquilaria sinensis excised twigs to produce sesquiterpenes of agarwood.In this study, the soild culture of endophytic fungal strain Fusarium solani A2 is in the Aquilaria sinensis, which was extracted by different solvent(PET and Et OAc) in turns. 7 compounds were isolated and purified byvarious chromatographic methods(silica gel, revese phase 18, Sephadex LH-20, preparative TLC, HPLC,recrystallization) from the PET layer and the Et OAc layer.Their structures were determined by NMR and MS spectroscopic data analyses, they were β-sitosterol(1),apigenin-7,4’-dimethylethers(2),6-methoxy-2-(2-phenylethyl)chromone(3),7,3’,4’-tri-O-methylluteolin(4),6-methoxy-2-[2-(3’-methoxyphenethyl)ethly]chromone(5),genkwanin(6), luteolin-7,4’-dimethylethers(7). Fimetariella rabenhorstii A20 was culture by malt extract medium. 8 compounds were isolated and purified.Their structures were identified.They were indazole(8), Methylphenol(9), globosumones C(10),cyclo(4-hydroxyl-Pro-Leu)(11),cylco(Pro-Tyr)(12),5α,8α-Epidioxyergosta-(22E,24R)-6,22-dien-3β-ol(13), 3β,5α-dihydroxy-(22E,24R)-ergosta-7,22-dien-6-one(14),(22E,24R)-ergosta-7,22-dien-3β,5α,6b-triol(15). Compound 1-6 are isolated from the genus Fusarium for the first time.Compounds 8-12, 14-15 are isolated from the Fimetariella rabenhorstii for the first time.Part of the compounds were tested for their cytotoxicities by SRB.The result shows that compound 2, 3, 4, 7, 10, 15 were demonstrated to inhibition against cell lines SF-268, MCF-7, NCI-H460, He PG-2.