Isolation and Identification of Endophytic Fungi from Aquilaria sinensis and Agarwood Induction
Objective:To isolate and purify the xylem fungi from Aquilaria sinensis induced by whole-tree agarwood-inducing technique(Agar-WIT),and screen the specific strains that effectively promoted the production of high-quality agarwood.Methods:The strains were isolated and purified by tissue separation method,and the fungi were identified by morphological characteristics combined with internal transcribed spacer(ITS)sequencing.Fungi solution was inoculated into A.sinensis for 4 months for functional verification.According to the agarwood standards in the Pharmacopoeia of the People's Republic of China(hereinafter referred to as the Chinese Pharmacopoeia,2020 edition),the effect of strain-induced agarwood was analyzed by thin layer chromatography,extract determination,agarotetrol determination and characteristic chromatogram.Results:A total of 48 strains of fungi were isolated from the xylem of A.sinensis,and the dominant strain was Fusarium sp.The thin-layer chromatogram of the agarwood induced by Colletotrichum sp.,A.niger and T.paraviridescens were consistent with those of the reference medicinal material.The extract content of agarwood induced by Fusarium sp.,A.niger and T.paraviridescens were more than 10%,and the difference was statistically significant when compared with that in the control group(P<0.05).The characteristic chromatograms of agarwood induced by 6 strains such as L.laosensis,A.niger and T.paraviridescens were in accordance with those of the reference medicinal materials,and the content of agarotetrol was more than 0.1%,higher than that in the control group(P<0.05).Conclusion:Endophytic fungi isolated from the xylem of A.sinensis could induce the formation of agarwood,among which A.niger and T.paraviridescens induced the formation in a more quickly way.The quality of resin obtained by agarwood formation conformed to the standards in the Chinese Pharmacopoeia(2020 edition).
国家科技期刊平台
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