东北膜荚黄芪内生真菌的分离及其活性代谢物分析  

Isolation of endophytic fungi from Astragalus membranaceus (Fisch.) Bge. and analysis of their secondary metabolites

作　　者：郑雪[1] 徐慧超[1] 李伟[1] 郑春英[1] 

机构地区：[1]微生物黑龙江省高校重点实验室,黑龙江大学生命科学学院,哈尔滨150080

出　　处：《中国新药杂志》2017年第12期1376-1381,共6页Chinese Journal of New Drugs

基　　金：黑龙江省自然科学基金(面上项目)(H2015003);哈尔滨市科技局优秀学科带头人项目(2014RFXXJ094)

摘　　要：目的:研究北芪内生真菌及其活性代谢产物,旨在深入开发新资源。方法:采用平板分离法从北芪中分离内生真菌,利用滤纸片法对内生真菌活性代谢物进行抑菌活性测定,以芒柄花素及北芪生药为对照,采用HPLC法对北芪内生真菌活性代谢物进行检测分析,同时对筛选出的目的菌株结合形态特征观察及r DNA ITS序列分析进行菌种鉴定。结果:在所得北芪内生真菌中,有25株内生真菌发酵液对11株试验菌表现出不同的抑菌活性,占总分离菌株的83.33%;HPLC分析表明,1株内生真菌发酵产物与北芪生药样品在相同保留时间内有相应的色谱峰出现,其中菌株HZ13经鉴定为链格孢属菌(Alternaria alternata)。结论:北芪内生真菌的发酵液具有较好抑菌活性,其中内生真菌HZ13发酵产生北芪活性成分或类似化合物,这为利用内生菌发酵生产北芪活性成分提供了新思路和途径。Objective： To investigate the endophytic fungi from Astragalus membranaceus（ Fisch.） Bge.and their secondary metabolites in order to find novel resources. Methods： Endophytic fungi were isolated from Astragalus membranaceus（ Fisch.） Bge. by using the method of plate cultivation,and the antimicrobial activities of their secondary metabolites were determined by filter paper method. Meanwhile,with formononetin and Astragalus membranaceus（ Fisch.） Bge. real samples as reference substances,the secondary metabolites of these endophytic fungi were analyzed by HPLC,and the target endophytic fungi were identified by r DNA ITS sequence homology analysis combined with their physiological and biochemical characteristics. Results： The fermentation broths of 25 strains of endophytic fungi（ 83. 33% of the total isolated strains） showed different antimicrobial activities,and the HPLC analysis indicated that there were the same corresponding chromatographic peaks appeared in the fermentation broths of HZ13 as its host plants at the same retention times,then the HZ13 strain was identified as Alternaria alternata. Conclusion： Diverse antimicrobial natural products exist in the secondary metabolites of endophytic fungi from Astragalus membranaceus（ Fisch.） Bge.,and HZ13 can produce similar or new active metabolites as its host.These findings provided a new idea and approach for using endophytic HZ13 to ferment and produce the components of Astragalus membranaceus（ Fisch.） Bge.

关 键 词：黄芪 内生细菌 抑菌活性 活性代谢物 

分 类 号：R284.1[医药卫生—中药学] R945[医药卫生—中医学]