摘要：内生真菌 (Endophytic Fungi) 是指寄生在植物组织体内并没有对其产生明显的病害的真菌。近来，人们发现植物内生真菌有重要的实用价值，具有抗肿瘤、抗病毒、抗菌、抗氧化等特性。目前，对植物内生真菌的分离研究已经成为微生物领域的一个研究趋势。51861

本课题从扬州、徐州采取了植物标本进行分离，采用组织分离法分离出多种植物内生真菌，对其进行纯化，并进行发酵培养后。利用抗肿瘤体外细胞毒筛选模型(MTT法)对内生真菌的代谢产物进行活性检测，检测其代谢产物的抗肿瘤性，筛选出具有高抗肿瘤活性的菌株，然后提取目标内生真菌的DNA，扩增其18SrDNA 测序，在数据库进行Blast，并结合菌丝形态和产孢结构鉴定目标内生真菌的分类地位。 根据实验结果得出，分离出的20株植物内生真菌，在不同的部位能够分离出不同的内生真菌。根据MTT结果显示一株植物冬青白—细极链格孢对A549有明显的抑制作用。

毕业论文关键词：内生真菌； 抗肿瘤性； 药用植物 

Separation and purification of endophytic fungi from a holly and identification of anti-tumor activities

Abstract: Endophytic Fungi refers to the parasitic plant tissue in the body and not obvious on the fungal diseases. Recently, it was found that endophytic fungi have important practical value, has antitumor, antiviral, antibacterial, antioxidant and other properties. At present, the research on the separation of endophytic fungi has become a research trend in the field of microbiology.

  This topic from picking from Yangzhou, Xuzhou plant specimens using tissue separation process in a variety of endophytic bacteria was isolated, purified, and after fermentation cultivation, use of antitumor cells in vitro drug screening model (determined by MTT method) was carried out on the metabolites of endophytic fungi activity detection, detection of its anti- tumor metabolites, screened strains with high activity, and extract the target DNA of endophytic fungi, expanded its 18 srDNA sequencing, Blast in the database, and combined with morphology and spores hyphae appraisal target the classification status of endophytic fungi. According to the experimental results concluded that 20 strains isolated endophytic fungi, in different parts of the isolated endophytic fungi of different. According to the determined by MTT showed a plant Holly fine white - a chain, spore has obvious inhibitory effect on A549.

Key words: endophytic fungi; antitumor; medicinal plants

目  录

摘  要 1

Abstract 1

引 言 1

1 材料与方法 2

1.1 样本 2

1.2 培养基 2

1.3试剂与仪器 2

2 实验方法 2

2.1 内生真菌的分离 2

2.2 MTT检验 5

2.3提取DNA 6

2.4 PCR检验 7

3 实验结果 7

3.1 植物内生真菌分离结果 7

3.2 菌丝形态分析 7

3.3 MTT检验结果 8

3.4 测序结果 9

讨  论 10

致  谢 11

参考文献 一株冬青内生真菌的分离纯化及鉴定及其抗肿瘤活性研究:http://www.751com.cn/shengwu/lunwen_55637.html