丹参中麦角甾醇高产内生真菌的鉴定及分析  

Identification and analysis of an endophytic fungus isolated from Salvia miltiorrhiza that produce ergosterol

作　　者：李秀清[1,2] 舒志恒[1] 明乾良[1] 秦路平[1] 郑承剑[1] 

机构地区：[1]第二军医大学药学院生药学教研室,上海200433 [2]华东师范大学生态与环境科学学院生态学教研室,上海200241

出　　处：《药学服务与研究》2016年第6期448-451,共4页Pharmaceutical Care and Research

基　　金：科技部863计划(2014AA020508);国家自然科学基金资助项目(81473301);上海市教委晨光计划(13CG40);上海市优秀青年医学人才培养计划(XYQ2013100)

摘　　要：目的:从药用植物内生真菌中筛选高产麦角甾醇的菌株。方法:使用马铃薯琼脂培养基从丹参中分离纯化内生真菌,马铃薯液体培养基用于菌株的发酵。基于内转录间隔区(internal transcribed spacer,ITS)序列分析进行菌株鉴定,利用HPLC仪,对比对照品的保留时间与紫外可见(ultraviolet-visible,UV)光谱来分析确定目标化合物。结果:从丹参叶中共分离得到58株内生真菌,合并为42个形态型,并筛选得到一株麦角甾醇高产目标菌株。该菌株通过形态和ITS序列分析,鉴定为球状茎点霉Phoma glomerata D14。通过与对照品的对比,确定丹参内生真菌D14可产生麦角甾醇,得率为10mg/g。结论:从丹参内生真菌中筛选得到一株麦角甾醇高产目标菌株球状茎点霉P.glomerata D14,提示该内生真菌在低成本、简便、快速获取满足医药用途的麦角甾醇需求方面具有良好的工业和科研意义。Objective： To screen candidate strains that produced ergosterol from endophytic fungus isolated from medicinal plants. Methods： Potato dextrose agar （PDA） medium was employed for the isolation and culture of fungal endophytes from Salvia rniltiorrhiza, while potato dextrose broth （PDB） medium was used for fungal fermentation. The candidate fungi was identified based on internal transcribed spacer（ITS） analysis and the target compound was elucidated as ergosterol, compared with the retention time and UV profile of authentic standard by HPLC method. Results.. A total of 58 endophytic fungal strains were isolated from the leaf of S. miltiorrhiza and sorted to 42 morphologies. The fungi produced ergosterol was identified as Phoma glomerata D14 based on its morphology and ITS analysis. Ergosterol in the mycelia was identified by HPLC method with a yield of 10 mg/g. Conclusion： An endophyte P. glornerata D14 which could produce ergosterol with high yield was isolated from S. miltiorrhiza. This result indicates that endophytic fungi has significant industrial and scientific potential to meet pharmaceutical demands for ergosterol in a cost-effective and easily accessible way.

关 键 词：内生真菌 球状茎点霉 麦角甾醇 丹参 微生物发酵 

分 类 号：Q93[生物学—微生物学]