蛇足石杉内生真菌产石杉碱甲含量检测及真菌的鉴定

Detection and identification of high producing huperzine A by an endophytic fungus from Huperzia serrata

目的从野生蛇足石杉中筛选高产石杉碱甲(HA)内生真菌。方法以野生蛇足石杉中分离获得的内生真菌为研究起点,采用TLC及HPLC法测定和分析并利用形态学及r DNA-ITS序列分析对高产HA菌株进行鉴定。结果初步形态鉴定内生真菌NSG-1为1株尖镰孢(镰刀菌)属真菌,经rDNA-ITS序列分析鉴定为尖镰孢NSG-1株。该菌株发酵液产HA为11.1mg/L,其产HA能力较以往研究菌株产量高。结论野生蛇足石杉中可获得高产HA内生真菌,其代谢产物中HA含量有待通过菌种诱变和改造来适应工业化生产。

Objective To screen endophytic fungal strains isolated from wild Huperzia serrata which can highly produce huperzine A. Methods The strain producing huperzine A was identified through thin layer chromatography（TLC）and high-performance liquid chromatography（HPLC）with authentic huperzine A. The fungus was identified according to its morphological characteristics and nuclear ribosomal DNA ITS sequence. Results The strain was identified as Fusarium oxysporum NSG-1 according to its morphological characteristics and nuclear ribosomal DNA ITS sequence. The amount of huperzine A produced by the fermentation liquor of this endophytic fungus was quantified to be 1.11 mg/100 ml by HPLC,which was higher than that of previously reported endophytic fungi. Conclusion Endophytic fungus producing high huperzine A can be obtained from H. serrata. The production of huperzine A based on mutagenesis and transformation of the obtained strain may adapt to the industry production.