蛇足石杉中产石杉碱甲内生真菌的分离和鉴定

Isolation and identification of endophytic fungus producing Huperzine A from Huperzia serrata

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。

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 HupA producing endophyte (NSH-5) was identified as Fusarium verticillioides according to its morphological characteristics and an analysis based on nuclear rDNA-ITS sequences. The amount of HupA produced by NSH-5 was quantified to be 11.76 mg/100 mL on liquid culture basis by HPLC, which with hereditary stability after 20 generations. [Conclusion] Strain NSH-5 could produce HupA. Isolation of such a fungus may provide a promising alternative approach to producing HupA.