甘草内生菌的鉴定及总黄酮 总皂苷的含量测定

Identification and Determination of Total Flavonoids and Total Saponins Contens in Endophytes from Glycyrrhiza Uralensis

目的对甘草内生菌进行形态学及分子生物学鉴定,并建立甘草内生菌代谢产物中总黄酮和总皂苷含量测定方法,对其含量进行测定。方法根据甘草内生菌的菌落、菌体形态进行初步鉴定,进一步采用16SrDNA、ITS测序进行分子生物学鉴定。采用紫外分光光度法,以甘草苷、甘草酸铵为对照品,在波长334、250nm处测定,计算甘草内生菌代谢产物中总黄酮和总皂苷的含量。结果 16株内生菌中,14株为内生细菌,2株为内生真菌。分子生物学鉴定结果表明,13株甘草内生细菌属于芽孢杆菌属,1株内生细菌属于乳杆菌目气球菌科,2株内生真菌属于曲霉菌科曲霉属。16株甘草内生菌代谢物中总黄酮和总皂苷含量存在差异,总黄酮含量在1.13~2.24mg·g^(-1),总皂苷含量在50.15~71.34mg·g^(-1)。结论紫外分光光度法测定甘草内生菌代谢产物中总黄酮、总皂苷含量的方法稳定、重复性好、可靠性高。

Objective To identify sixteen strains of endophytes from Glycyrrhiza uralensis by morpholoqical and molecular biological methods,and to establish a method for determination of total flavonoids and total saponins in the metabolites of endophytes.Methods Endophytes from Glycyrrhiza uralensis were preliminarily identified with the morpholoqical method before being identified based on 16 S rDNA or ITS sequence.With liquiritin and ammonium glycyrrhetate as the control,the contents of total flavonoids and total saponins in the metabolites of endophytes from Glycyrrhiza uralensis were determinedby ultraviolet spectrophotometry at a wavelength of 250 and 334 nm respectively.Results Fourteen strains of endophytes were Bacteria,and two strains were Eukaryota.According to molecular biology,thirteen strains of endophytes were Bacteria Bacillus,one strain was Aerococcaceae Aerococcus.,and one strain was Aspergillaceae Aspergillus.There was difference in the contents of total flavonoids and total saponins between the metabolites of the sixteen strains of endophytes fromGlycyrrhiza uralensis.The content of total flavonoids was 1.13-2.24 mg·g-1,while that of total saponins was 50.15-71.34 mg·g-1.Conclusion Ultraviolet spectrophotometry can be used to measure the contents of total flavonoids and total saponins in the metabolites of endophytes fromGlycyrrhiza uralensis.This method is simple and quick with good stability,repeatability and accuracy.