产黄酮银杏内生真菌的分离与鉴定

摘要

本研究利用组织块培养法从健康的银杏根、茎中分离得到116株内生真菌.通过显色反应、高效液相色谱(HPLC)等方法对内生真菌的代谢产物进行分析,最终筛选出2株能够产黄酮的内生真菌,利用分光光度法测定其发酵液中总黄酮产量均达到20 mg/L以上.形态学特征和真菌rDNA间隔序列(Internal Transcribed Spacer,ITS)的系统进化分析结果表明两株菌分别属于青霉属和毛霉属,其中青霉属为产黄酮银杏内生真菌的首次报道.本研究为利用微生物发酵生产黄酮类化合物奠定了良好的基础.%Many studies have shown that endophytes can co-evolution with host,and can produce similar or identical physiologically active substances as host.Flavonoid is one of most active compounds in extracts of Ginkgo biloba.In the present study,the method of tissue culturing was used to isolate endophytic fungi from health ginkgo roots and stems.After repeated separation and purification,116 Ginkgo endophytes were isolated.Metabolites of these fungi was preliminarily analyzed by color reaction and high performance liquid chromatography (HPLC),and two flavonoids producing endophytes were finally screened out.The production of flavonoids by these two strains were all more than 20 mg/L.These two endophytic fungi strain were identified as Mucor spp.and Penicillium spp.by morphology characteristics and Internal Transcribed Spacer (ITS) sequences analysis,and Penicillium spp.was the first report in the flavonoid producing endophytic fungus from G.biloba.This study laid a good foundation for the production of flavonoids by microbial fermentation,and these two strains can be induced to improve the production of flavonoids by mutation breeding before they are used to produce flavonoids by fermentation.