黄檗植物产小檗碱的内生真菌的分离鉴定

Isolation and Identification of Producing Endophytic Fungi of Berberine from the Plant Phellodendron amurense

[目的]探索黄檗植物产小檗碱的内生真菌的分离与鉴定。[方法]从黄檗中分离内生真菌,采用改良的碘化铋钾试剂及薄层层析法对其代谢产物进行分析,对可能产生小檗碱的菌株进行扩大培养,然后对培养后提取物采用KBr涂片法进行红外光谱检测,并以氘代甲醇为溶剂进行核磁共振氢谱检测,最后进行初步形态学分类。[结果]从黄檗中分离得到13株内生真菌,其中菌株BBH6可能产生小檗碱;通过扩大培养以及红外光谱检测和核磁共振氢谱检测,最终确定内生真菌BBH6能够产小檗碱;该菌株菌丝发达,分生孢子暗色,多细胞,由横纵隔膜分成砖格状,依据真菌形态分类法,该菌株应归属于真菌门的交链孢霉属。[结论]该研究为药用植物内生真菌的研究提供了参考依据和借鉴。

[ Objective ] The research aimed to explore the isolation and identification of producing endophytic fungi of berberine from the plant Phellodendron amurense. [ Method] The endophytic fungi were isolated from P. amurense and their metabolites were analyzed by improving bismuth potassium iodide reagent and thin layer chromatography. The strains, which may produce berberine, were used in enlarged culture, then the infrared spectrum detection was performed on the extracts produced in the enlarged culture by KBr smearing method and the NMR-H spectrum detection was performed with CD30D as solvent. Finally, they were classified by morphologically preliminarily. [ Result] Thirteen strains of endophytic fungi were isolated from P. amurense, among them the strain BBH6 may produce berberine. It was confirmed that the endophytic fungus BBH6 could produce berberine finally through enlarged culture ,infrared spectrum detection and NMR-H spectrum detection. BBH6 had developed myeelia,its conidia were dark color,multi-cellular and divided into muriforms by cross and vertical membranes. On the basis of mor- phological classification of fungi, BBH6 should be belong to Alternaria in Eumycota. [ Conclusion ] The research provided reference and basis for researching the endophytic fungi in medicinal plants.