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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 To isolate endophytic fungi producing gentiopicroside from roots, stems and leaves of medicinal plant Gentiana straminea, in order to find microbial resources that can substitute for plants and thus to better protect the resources of wild Gentiana straminea. Methods Endophytic fungi were isolated from roots, stems and leaves of Gentiana straminea from Aba, Sichuan Province by tissue culture using PDA medium and analyzed by high performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) ) Endophytic fungus fermentation products were tested. Finally, morphological and molecular biological means of endophytic fungi screening strains identified. Results A total of 10 endophytic fungi were isolated from roots, stems and leaves of Gentiana straminea from Aba, and the n-butanol extract of one of the endophytic fungi GJ-01 was purified by high performance liquid chromatography ) And liquid chromatography-mass spectrometry (LC-MS). The results showed that there was gentiopicroside in its secondary metabolites, which was 4.16mg / L. Morphological and molecular biological methods identified, confirmed that endophytic fungi GJ-01 Fusarium. Conclusion An endophytic fungus GJ-01 producing gentiopicroside was isolated from Gentiana straminea from Aba, which could be used as a new microbial resource of gentiopicroside.