摘要：近几年，研究者们开始探索新途径来研制新药，尝试从天然产物中筛选生物活性成分。同时研究人员们也已经开始注意到植物可被看作是内生真菌的储存库，大部分内生真菌能够合成具有新活性的代谢产物，可以为植物提供对病原体的防御，并且这些化合物中已经有一部分被证明为有新的药用价值，如抗菌、抗肿瘤、抗氧化等。

本课题是从采摘自无锡，安徽等地的植物标本中分离出多种植物内生菌，对其进行纯化，并进行发酵培养，用MTT法检测其代谢产物的抗肿瘤活性。从而来发现我们日常生活周围的植物中也有含抗肿瘤活性的内生真菌的存在，一些不起眼的植物也同样具有研究价值。51842

毕业论文关键词：花生；内生真菌；抗肿瘤;

Isolation Identification of Endophytes in and the Antitumor Activity Studies

Abstract: In recent years, screening bioactive ingredients from natural products has become an effective way of developing new drugs. At the same time, researchers have also started to notice that plants can be regarded as a repository of endophytic fungus which are capable of synthesizing new active metabolites and providing defense against pathogens for plants. Several of these compounds have been shown to have a new medicinal value such as there proved to be part of the new medicinal value, such as antitumor, antiviral, antibacterial, anti-oxidation. 

Firstly, endophytes were isolated from plants specimens picked from Wuxi and Anhui. Secondly, these endophytes were purified and fermented to detect their anti-tumor activity. We found that there are also endophytic fungi containing anti-tumor activity in the plants in our daily life. Some humble plants are of great value. 

Key words：Peanut; Endophytic fungus; Anticancer;目录

1 引言 1

1.1内生真菌的发展 1

1.2菌种 1

1.3 内生真菌的抗肿瘤活性 1

2 材料与方法 2

2.1 实验材料 2

2.1.1 样品 2

2.1.2 仪器 2

2.1.3 材料与试剂 2

2.1.4 培养基    2

2.2 实验方法 2

2.2.1 植物内生真菌的分离 2

2.2.2 菌种纯化 4

2.2.3菌种保藏 4

2.2.4 菌种发酵及过滤 4

2.2.4.1菌种发酵 4

2.2.4.2  发酵液体过滤 5

2.2.5 MTT法检测内生真菌代谢产物的抗肿瘤活性 5

2.2.5.1材料 5

2.2.5.2细胞培养 5

2.2.5.3细胞复苏 5

2.2.5.4细胞传代 5

2.2.5.5细胞冻存 6

2.2.5.6 MTT实验 6

2.2.5.7 计算IC50 6

2.2.6菌种分子鉴定 6

2.2.6.1提取DNA 6

2.2.6.2 PCR测序 7

2.2.7菌种形态鉴定 7

3 结果与分析