蛇足石杉内生真菌产石杉碱甲含量检测及真菌的鉴定

摘要

Objective To screen endophytic fungal strains isolated from wild Huperzia serrata which can highly produce hu⁃perzine A. Methods The strain producing huperzine A was identified through thin layer chromatography(TLC)and high-perfor⁃mance liquid chromatography(HPLC)with authentic huperzine A. The fungus was identified according to its morphological character⁃istics 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 li⁃quor of this endophytic fungus was quantified to be 1.11 mg/100 ml by HPLC,which was higher than that of previously reported endo⁃phytic fungi. Conclusion Endophytic fungus producing high huperzine A can be obtained from H. serrata. The production of huper⁃zine A based on mutagenesis and transformation of the obtained strain may adapt to the industry production.%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rDNA-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11.1 mg/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。