蛇足石杉中产石杉碱甲内生真菌的分离和鉴定-微生物学报.PDF

微生物学通报 Sep. 20, 2017, 44(9): 2153−2160 Microbiology China /wswxtbcn tongbao@ DOI: 10.13344/j.microbiol.china.170160 研究报告 蛇足石杉中产石杉碱甲内生真菌的分离和鉴定 1* 1 1 1 1 1 1 2 韩文霞 李伟泽 李小峰 张寒 杨时珍 杜江凤 景欢 程传彬 (1. 710021) (2. 050083) 摘 要：【目的】从野生蛇足石杉(Huperzia serrata) 中分离筛选产石杉碱甲的内生真菌。【方法】 采用薄层层析及高效液相色谱法对内生真菌代谢产物进行测定和分析以期分离获得产石杉碱 甲菌株，运用形态及ITS 序列分析方法对产石杉碱甲菌株进行鉴定，并利用连续传代方法考察 菌株遗传稳定性。【结果】经筛选获得一株产石杉碱甲内生真菌NSH-5 ，经形态学鉴定及 ITS 序列分析鉴定为轮枝镰孢菌(Fusarium verticillioides) ，其石杉碱甲产量为11.76 mg/100 mL，菌 株经20 次连续传代后遗传稳定。【结论】NSH-5 菌株为一株具有产石杉碱甲能力的轮枝镰孢菌， 该菌株的发现为生物合成石杉碱甲提供了新的菌种资源。 关键词：石杉碱甲，薄层层析法，高效液相色谱法，轮枝镰孢菌 Isolation and identification of endophytic fungus producing Huperzine A from Huperzia serrata HAN Wen-Xia1* LI Wei-Ze1 LI Xiao-Feng1 ZHANG Han1 YANG Shi-Zhen 1 DU Jiang-Feng1 JING Huan1 CHENG Chuan-Bin2 (1. Xi’an Medical University, Xi’an, Shaanxi 710021, China) (2. Shijiazhuang Institute of Mechanized Infantry, Shijiazhuang, Hebei 050083, China) Abstract: [Objective] To screen and isolate endophytic fungal strains producing Huperzine A (HupA) from wild Huperzia serrata . [Methods] HupA producing endophytes were identified through thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC), and the identification of the fungus with relatively a high HupA productivity was performance based its morphological characteristics and nuclear rDNA-ITS sequences. [Results] One