摘要：目的：从药用植物白芍中分离内生真菌并进一步研究其次生代谢产物。 方法：在无菌条件下从白芍各组织块中分离获得内生真菌并通过ITS序列测定法进行菌种鉴定。采用对峙培养法进行菌株的抗菌活性初筛定，通过代谢产物的含量测定，薄层层析法选取代谢产物含量高、代谢产物丰富或具有抗菌活性的内生真菌作为目标菌株。目标菌株大量发酵培养获得粗提物，再经薄层层析、中压液相层析、凝胶柱Sephadex LH-20、制备薄层色谱、重结晶等方法分离纯化获得单体化合物。采用核磁共振谱，质谱等鉴定单体化合物。结果：从白芍茎中分离获得6种内生真菌，筛选获得目标菌株为角担子菌属真菌BS-24，从其代谢产物中分离获得2个化合物，经波谱数据分析鉴定出1个化合物：N-苯基-2-萘胺。结论：白芍内生真菌的种类繁多、代谢产物较为丰富，值得进一步的研究。89188

Abstract: Objective:Isolation of endophytic fungi in medicinal plant Radix Paeoniae Alba and then study their secondary metabolites。 Methods：Endophytic fungi were isolated from the tissues of Radix Paeoniae Alba under sterile conditions and identified by ITS sequencing。 The antimicrobial activity of the strain was tested by confrontation culture method。 The content of metabolites was tested by thin layer chromatography, and the endophytic fungi with rich metabolites or antibacterial activity were selected as target strains to obtain crude extracts。 And then they were separated and purified with thin layer chromatography, medium pressure liquid chromatography, gel column Sephadex LH-20, preparative thin layer chromatography and recrystallization to get monomeric compounds。 The purified compounds were identified by nuclear magnetic resonance spectroscopy, mass spectrometry etc。 Results: Six strains of endophytic fungi were isolated from R。Paeoniae Alba, Metabolites of the target strain Ceratobasidum sp。 BS-24 were separated to get two pure compounds。 One compound was identified as N-phenyl-2-naphthylamine through spectroscopic methods。 Conclusion: The abundant of endophytic fungi as well as their metabolites in Radix Paeoniae Alba make it worthy of further study。源Y于U优I尔O论P文W网wwW.yOueRw.com 原文+QQ75201-8766

毕业论文关键词：白芍； 内生真菌； 代谢产物；抗菌活性

Keyword: Radix Paeoniae Alba； Endophytic Fungi；Metabolites；Antibacterial activity

目    录

摘    要 2

引言 4

白芍研究概况 4

内生真菌研究进展 4

前期基础 4

1 实验材料 5

1。1 仪器及试剂 5

1。2 药材来源 6

1。3 抗菌实验用指示菌 6

1。4 常用培养基 6

2 实验方法 7

2。1 内生真菌培养、分离及纯化 7

2。2 内生真菌鉴定 8

2。2。1 DNA的提取 8

2。2。2 PCR扩增、测序 8

2。3 目标菌株筛选 8

2。3。1 代谢产物含量初测 8

2。3。2 代谢产物丰富度测定 8

2。3。3 抗菌活性初筛定 9

2。4 目标菌株发酵培养