一株产龙胆苦苷的麻花秦艽内生真菌的分离鉴定

Isolation and identification of an endophytic gentiopicrin-producing fungus obtained from Gentiana straminea Maxim

目的从药用植物麻花秦艽的根、茎和叶中分离出可产龙胆苦苷的内生真菌菌株,以寻找可替代植物的微生物资源,从而可以更好地保护野生麻花秦艽资源。方法用PDA培养基,采用组织分离法从四川阿坝红原麻花秦艽的根、茎和叶中分离内生真菌,并通过高效液相色谱(HPLC)和液相色谱-质谱联用(LC-MS)对内生真菌的发酵产物进行检测。最后用形态学和分子生物学手段对筛选出的内生真菌进行菌株鉴定。结果从阿坝红原麻花秦艽的根、茎和叶中总共分离得到了10株内生真菌,其中的一株内生真菌GJ-01的发酵液的正丁醇萃取物通过高效液相色谱(HPLC)和液相色谱-质谱联用(LC-MS)检测,结果表明其次级代谢产物中存在龙胆苦苷,含量为4.16mg/L。经形态学和分子生物学手段鉴定,确认内生真菌GJ-01为腐皮镰刀菌。结论从阿坝红原麻花秦艽中分离得到了一株产龙胆苦苷的内生真菌GJ-01,可作为一种新的产龙胆苦苷的微生物资源。

Objective In order to protect the wild resource of Gentiana straminea Maxim.,we isolated endophytic fungui from the root,stem and leaf of Gentiana straminea Maxim. growing in Aba Hongyuan,Sichuan province. Methods Endophytic fungi was isolated from the root,stem and leaf of Gentiana straminea Maxim. growing in Aba Hongyuan by the tissue separating method. These endophytic fungui＇s fermentation broth＇s n-butanol extracts were detected byhigh-performance liquid chromatography（ HPLC）and liquid chromatography-mass spectrometry（ LC-MS）. The screened endopytic fungui were then analyzed by the morphological traits and the flanking internal transcribed spacers sequencing（ ITS）. Results 10 endophytic fungi were isolated from Gentiana straminea Maxim. growing in Aba Hongyuan by the tissue separating method. Among them,the strain GJ-01 was found to produce gentiopicroside with a yield of 4. 13 mg / L,detected by HPLC and LC-MS. According to the morphological traits and the analysis of the ITS,the strain GJ-01 was grouped into Fusarium solani strain. Conclusion One gentiopicrin-producing endophytic fungus was isolated from the root of Gentiana straminea Maxim. and this strain has the potential to become a new microbial resource for producing gentiopicroside.