摘要通过萃取、薄层层析等方法将一株内生真菌代谢产物的抑菌活性成分初步分离。用微量肉汤稀释法验证此内生真菌菌株发酵提取物的抑菌活性，再依次用石油醚、乙酸乙酯和正丁醇进行萃取，分别验证各相提取物的抑菌活性，结果表明抑菌物质集中在乙酸乙酯相和正丁醇相。将提取物点样于薄层层析板上，用四种展层剂展开，结果显示，当点样量为200ul，用五号展层剂（正丁醇饱和的0。5mol×L-1，pH7。0的磷酸缓冲溶液）展开时，薄层板上2-3cm处的硅胶粉中含有抑菌活性成分，将其刮下后放置于涂布携带NDM-1 β-内酰胺酶基因重组质粒的大肠杆菌的LB平板上过夜培养后，其周围出现抑菌圈，推测抑菌活性成分的Rf值为：0。33-0。5。本研究为进一步分离抑菌活性成分奠定了基础。72222

该论文有图3幅，表4个，参考文献15篇。

毕业论文关键词：内生真菌  薄层层析  抑菌活性成分 抑菌

A Preliminary Study on the Isolation of Endophytic Fungi from Endophytic Fungi

Abstract In the present study, the antibacterial components from the metabolites of an endophytic fungus was separated preliminarily by solvent extraction method and thin layer chromatography。 The crude extract of the fermentation broth of the endophytic fungus was obtained。 The antibacterial activity of the extract was determined by the micro-broth dilution method, and then further extracted with petroleum ether, ethyl acetate and n-butanol successively。 The antibacterial activity of each extract phase was tested。 The results showed that the antibacterial substances concentrated in ethyl acetate and n-butanol。 The extract was sampled into silica gel plate and developed with four kinds of developing agent。 The result showed that the antibacterial components reached the spot 2-3cm from the sampling site when 200ul of samples was spotted with No。 5 developing agent, which was proved by the inhibition zone appeared after the silicon powder placed on the LB plate coating suspension of Escherichia coli carrying recombinant plasmid with NDM-1 β-lactamase gene and cultured overnight。 It suggested that the Rf value of the antibacterial components was range 0。33 to 0。5。This study laid a foundation for further separation of antibacterial activity。

Key Words: Endophytic fungi  Thin layer chromatography  Antimicrobial active ingredient

目录

摘  要 Ⅰ

Abstract Ⅱ

目录 Ⅲ

图清单 Ⅳ

表清单 Ⅳ

引言 Ⅴ

一 材料与方法 6

1。1菌株 6

1。2 仪器 6

1。3 设备 6

1。4 实验材料 6

1。5 实验方法 7

二 结果与分析 9

三 讨论 12

参考文献 14

致谢 15

图清单

图序号 图名称 页码

图1 1号展层剂5ul抑菌结果 11

图2 2号展层剂5ul抑菌结果 11

图3