产哈巴苷和哈巴俄苷玄参内生真菌的分离及鉴定

从玄参根、主茎、侧枝、叶片中分离得到内生真菌,采用高效液相和液质联用的方法对内生真菌的代谢产物进行分析,筛选能够产哈巴苷或哈巴俄苷的内生真菌,根据菌株的形态学特征,结合分子生物学的系统进化分析进行菌株鉴定。结果表明从玄参中共分离得到内生真菌210株,合并为46个形态型,分属于2纲、9目、13科、17属。HPLC初步筛选出2株内生真菌能够分别产哈巴苷和哈巴俄苷,LC-MS进一步验证得到了能产哈巴俄苷的目标菌株ZJ17,鉴定为Alternaria alternate;以及能产哈巴苷的目标菌株ZJ25,鉴定为A. gaisen。

Isolation and identification of endophytic fungi producing harpagoside and harpagide from Scrophularia ningpoensis

The endophytic fungi from root,main stem,branch and leaf of Scrophularia ningpoensis were isolated from Zhejiang,whether these strains could yield harpagide or harpagoside were tested by HPLC and LC-MS. According to the morphological characteristic and the similarity of the nucleotide sequence of internal transcribed spacer( ITS) between r DNAs,the strains producing harpagide or harpagoside were identified. The results showed that 210 strains were isolated from the samples,which were classified into 9 orders,13 families and 17 genera by morphological study. Harpagide was detected in endogenous fungi ZJ17 and harpagoside was detected in endogenous fungi ZJ25 by HPLC coupled with LC-MS. ZJ17 was identified as Alternaria alternate and ZJ25 was identified as A.gaisen by its morphology and authenticated by ITS( ITS4 and ITS5 regions and the intervening 5. 8 S rDNA region).