茶树内生真菌次生代谢物检测与分析
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摘要
本论文以豫南地区茶园的茶树内生真菌为研究对象,在春、夏、秋、冬四季更替过程中,采用不同培养基对各季节茶树内生真菌进行分离培养和鉴定,共得到内生真菌9科16属27种,其中刺盘孢菌属(Colletotrichum)、大茎点霉属(Macrophoma)、拟茎点霉属(Phomopsis)等3属内生真菌在各个季节中均有分布且分离率都较高,为茶树优势内生真菌。研究表明,季节不同,茶树内生真菌种数明显不同,表现为春季最多,夏季最少,且夏季、秋季、冬季、春季的内生真菌种数依次呈递增趋势。
运用不同方法和手段对茶树内生真菌次生代谢产物中茶多酚类、多糖类和油脂类化合物进行了检测与分析。(1)采用薄层析法、酒石酸铁比色法及毛细管电泳检测法对茶多酚类次生代谢物进行检测,三种方法检测结果一致,即无论是发酵液或是菌丝内均产生茶多酚类物质,说明茶树内生真菌可以产生与宿主植物相同或相似次生代谢物；(2)用斐林试剂对茶树叶栖内生真菌7个菌株的多糖类物质检测发现,各菌株检测液均产生砖红色沉淀,说明茶树内生真菌在生长发育过程中可以在细胞外产生多糖类次生代谢物,且这一性状具有普遍性,并且测得QJ-Mac③菌株次生代谢产物溶液中,糖含量平均值为0.00137mg/mL;(3)通过对480株茶树内生真菌的产油性状进行显微形态学观察,结果发现,产油内生真菌有371株,占总菌株数77.3%,其中高产油内生真菌69株,占18.6%。表明,供试的绝大多数真菌种类均产生油脂,且茶树内生真菌产油性状与真菌种类密切相关,高产油内生真菌集中存在于拟茎点霉属(Phomopsis)及镰刀菌属(Fusarium)真菌菌株中。
This paper mainly treats tea tree's endophytic fungi in the southern Henan province as the research object. In the alternate seasons of the year, we use different culture medium on the seasonal isolation and identification of endophytic fungi in each season, finding that there are totally27species in16genera belonging to9families identified, of which Colletotrichum, Macophoma, Phomopsis, the distribution of these3genera of endophytic fungi existing in various seasons with higher separation rate, as advantage endophytic fungi. Research shows that in different seasons, types and volume of tea tree endophytic fungi is different:the most in spring, the least in summer; types and volume of the endophytic fungi in turn trend to increase in summer, fall, winter and spring.
Using different methods and means, tests and analyses are made in the tea polyphenols, polysaccharides and grease compounds of the tea tree endophytic fungi secondary metabolites.(1)Tea polyphenolic secondary metabolites are detected by thin layer chromatography, tartaric acid iron colorimetric method and the method of capillary electrophoresis. Three methods detection results are consistent, namely whether fermented liquid or hyphae, they can both produce tea polyphenols, meanwhile it also proves the viewpoint that tea tree's endophytic fungi can produce the same or similar secondary metabolites with host plants;(2) By Fehling reagent on the tea leaf tissue of endophytic fungi in seven target strains of Polysaccharides material detection, each target strains liquid produces brick red precipitate, illustrating that the tea endophytic fungi in the growth and development process in the extracellular polysaccharides can produce secondary metabolites of this trait, which is universal, and find that in the liquor of secondary metabolites of the strain QJ-Mac, sugar content average was0.00137mg/mL;(3) Microscopic morphological observation is operated through480strains of tea tree oil of endophytic fungi characters. The results show that371strains are oil-producing, accounting for77.3%of the total number of strains, of which69strains are high-yield with high oil, accounting for18.6%, which proves that most of the species can produce oil and tea tree oil endophytic fungi is closely related to fungi type; and yielding oil intensively exists in fungus Phomopsis and Fusarium fungal strains.
运用不同方法和手段对茶树内生真菌次生代谢产物中茶多酚类、多糖类和油脂类化合物进行了检测与分析。(1)采用薄层析法、酒石酸铁比色法及毛细管电泳检测法对茶多酚类次生代谢物进行检测,三种方法检测结果一致,即无论是发酵液或是菌丝内均产生茶多酚类物质,说明茶树内生真菌可以产生与宿主植物相同或相似次生代谢物；(2)用斐林试剂对茶树叶栖内生真菌7个菌株的多糖类物质检测发现,各菌株检测液均产生砖红色沉淀,说明茶树内生真菌在生长发育过程中可以在细胞外产生多糖类次生代谢物,且这一性状具有普遍性,并且测得QJ-Mac③菌株次生代谢产物溶液中,糖含量平均值为0.00137mg/mL;(3)通过对480株茶树内生真菌的产油性状进行显微形态学观察,结果发现,产油内生真菌有371株,占总菌株数77.3%,其中高产油内生真菌69株,占18.6%。表明,供试的绝大多数真菌种类均产生油脂,且茶树内生真菌产油性状与真菌种类密切相关,高产油内生真菌集中存在于拟茎点霉属(Phomopsis)及镰刀菌属(Fusarium)真菌菌株中。
This paper mainly treats tea tree's endophytic fungi in the southern Henan province as the research object. In the alternate seasons of the year, we use different culture medium on the seasonal isolation and identification of endophytic fungi in each season, finding that there are totally27species in16genera belonging to9families identified, of which Colletotrichum, Macophoma, Phomopsis, the distribution of these3genera of endophytic fungi existing in various seasons with higher separation rate, as advantage endophytic fungi. Research shows that in different seasons, types and volume of tea tree endophytic fungi is different:the most in spring, the least in summer; types and volume of the endophytic fungi in turn trend to increase in summer, fall, winter and spring.
Using different methods and means, tests and analyses are made in the tea polyphenols, polysaccharides and grease compounds of the tea tree endophytic fungi secondary metabolites.(1)Tea polyphenolic secondary metabolites are detected by thin layer chromatography, tartaric acid iron colorimetric method and the method of capillary electrophoresis. Three methods detection results are consistent, namely whether fermented liquid or hyphae, they can both produce tea polyphenols, meanwhile it also proves the viewpoint that tea tree's endophytic fungi can produce the same or similar secondary metabolites with host plants;(2) By Fehling reagent on the tea leaf tissue of endophytic fungi in seven target strains of Polysaccharides material detection, each target strains liquid produces brick red precipitate, illustrating that the tea endophytic fungi in the growth and development process in the extracellular polysaccharides can produce secondary metabolites of this trait, which is universal, and find that in the liquor of secondary metabolites of the strain QJ-Mac, sugar content average was0.00137mg/mL;(3) Microscopic morphological observation is operated through480strains of tea tree oil of endophytic fungi characters. The results show that371strains are oil-producing, accounting for77.3%of the total number of strains, of which69strains are high-yield with high oil, accounting for18.6%, which proves that most of the species can produce oil and tea tree oil endophytic fungi is closely related to fungi type; and yielding oil intensively exists in fungus Phomopsis and Fusarium fungal strains.
引文
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