摘要：内生真菌是指，一段时间，与宿主植物共同生存，但对植物组织没有明显伤害的真菌。这些经过自身代谢就可产生具有重要生物学及生理学作用的生物活性成分的内生真菌，它的植物来源注定它的次级代谢产物中包含的活性成分将产生很大的区别。近几十年来，人们发现这些具有抗肿瘤、抗菌、抗病毒、抗氧化等特性的内生真菌，对人类将产生重大影响，具有非常重要的资源价值。本课题是从采摘自徐州，扬州等地的植物标本中分离出多种植物内生菌，对其进行取样，表面灭菌 ，平板接种及室温培养 ，菌种纯化，菌种保藏 ，菌种发酵，过滤发酵液，MTT法检测内生真菌代谢产物的抗肿瘤活性，最后再进行菌丝形态鉴定 ，菌种DNA的提取 。最后确定菌种。实验结果显示，分离出的20株内生真菌，在它的不同部位，能分离出的菌不尽相同。分别生长于徐州和扬州，产生的菌也不一样。MTT结果表明，一株桂花C4的一毛壳菌的发酵产物对A549有明显抑制作用，效果最为显著。76474

毕业论文关键词：内生真菌 ，分离纯化 ，鉴定 ， MTT法，抗肿瘤活性 

Separation and purification of endophytic fungi from a sweet-scented osmanthus and identification of antitumor activitie 

Abstract: Endophytic fungus refers to a period of time, to live together with the host plant, but there is no obvious damage to the plant tissue fungi。 These can produce through their own metabolic endophytic fungi have important biological and physiological effects of biological active ingredient, the active ingredient of its plant-derived secondary metabolites destined it contained would have a great difference。 In recent decades, people have found that the anti-tumor, anti-bacterial, anti-viral, anti-oxidation characteristics of endophytic fungus, will have a significant impact on humans, has a very important resource value。 This issue is separate from picking from Xuzhou, Yangzhou and other places of plant specimens in a variety of endophyte, its sampling, surface sterilized, plated and incubated at room temperature, bacteria and purified, culture collections, fermentation bacteria filtered broth, MTT assay endophytic fungal metabolites anti-tumor activity, then finally mycelium morphological identification, bacteria DNA extraction。 Experimental results show that separated within 24 strains of endophytic fungi in different parts of it, can be isolated bacteria vary。 Were grown in Xuzhou and Yangzhou, bacteria produce not the same。 MTT results showed that the fermentation product of a C4 is a sweet-scented osmanthus Chaetomium of A549 significantly inhibited, the most significant effect。

Keywords: Endophytic fungus, separation and purification, identification, antitumor activity, MTT method

目   录

摘 要 Ⅰ

Abstract Ⅱ

引 言 1

1 材料与方法 2

1。1材料 2

1。1。1分离材料 2

1。1。2培养基 2

1。1。3试剂与仪器 2

1。2实验方法 2

1。2。1内生真菌的分离 3

1。2。2平板接种及室温培养 3

1。2。3菌种纯化 3

1。2。4菌种保藏 3

1。2。5菌种发酵 4

1。2。6过滤发酵液 4

1。2。7 MTT法测抗肿瘤活性