南京林业大学.doc

南京林业大学
本科毕业设计(论文)
题目:伞滑刃属线虫RAPD分子标记的筛选
学院:森林资源与环境学院
专业:树木与观赏植物保护
学号: 080101319
学生姓名: 汪菲菲
指导教师: 韩正敏
职称: 教授



摘要
松材线虫(Bursaphelenchus xylophilus)是松树的一种灾难性病害的病原生物,是引起松树萎蔫病的主要原因。拟松材线虫(B. mucronatus)作为伞滑刃属的线虫同松材线虫经常混生或单独出现在病树木上,两者形态极其相似,较难区分。因此对两种线虫的准确、快速鉴定工作被认为是松材线虫病防控的最重要的环节之一。本文采用RAPD技术对松材线虫和拟松材线虫进行了检测研究,从30个随机引物中筛选出较好的引物S60,对来自6个不同地区的线虫的种源进行检测,能较为清晰地看出松材线虫与拟松材线虫的区别。对进一步研究线虫种间与种内亲缘关系提供了研究基础。
关键词:松材线虫;拟松材线虫;RAPDDNA扩增;分子标记检测
Abstract
Pine wood nematode( xylophilus xylophilus) is the pathogen of pine wilt disease,and is a devastating disastrous of the pine tree of the disease pathogen creatures, which caused the main reason of disease of pine trees. B. Mucronatus mucronatusalso as the nematode belong to the genus of Bursaphelenchus , is often mixed with Bursaphelenchus xylophilus Bx or appears alone in the disease on the tree; . their Their forms are too extremely similar to distinguish each other little difficultly. So, tTo identify exactly and quickly is regarded as one of the most important part of the prevention and control for pine wilt disease. In this article, Bursaphelenchus xylophilus Bx and B. mucronatusBm were tested with the technology of RAPD. To test the provenances of nematodes from six areas with the better primer S60 that screened thirty random primers, we can easily distinguish between Bursaphelenchus xylophiluBx and B. mucronatus ,aBm. And the technology provides a foundation to discuss the relation between interspecies and inter-species of nematodes.
Key Words: Bursaphelenchus xylophilus Bx ,B. mucronatus Bm , RAPD ,molecular marker
目录
4
4
4
4
松材线虫与拟松材线虫的几种常用鉴别方法 8
松材线虫与拟松材线虫分子标记的意义 10
松材线虫与拟松材线虫的研究概况 10
常用DNA提取方法 10
几种常用分子检测技术 12
松材线虫与拟松材线虫RAPD分子标记研究进展 12
实验过程应用主要原理 13
实验流程图 13
常用DNA提取方法 13
几种常用的分子检测技术 14
14
14
14
14
1