毕业设计（论文）-利用ISSR分子标记对39份莲藕种质的遗传多样性分析.doc

本科毕业论文设计 题　　目 利用ISSR分子标记对39份莲藕 种质的遗传多样性分析 姓 名 × × × 指导教师 × × × 2010年5月18日 利用ISSR分子标记对39份莲藕种质的遗传多样性分析 摘要：本研究首先优化了莲藕DNA的提取方法，采用正交设计方案对莲藕ISSR-PCR反应体系从4因素3水平进行了优化，用优化后的反应体系对引物进行了筛选，然后用得到的引物对39份莲藕DNA样品予以扩增，并对莲藕的遗传多样性进行了系统分析。通过正交实验得到了莲藕PCR扩增的最佳反应体系，利用该体系从设计的18条引物中筛选出了8条适合莲藕ISSR-PCR扩增的引物，共扩增出59条DNA条带，其中多态性条带37条，多态比率（PPT）为62.71%，平均每条引物扩增出7.36条带。利用NTSYS-pc软件对扩增的莲藕的遗传多样性分析表明，39份莲藕的遗传系数在0.63-0.98之间，用UPGMA聚类分析，在遗传系数0.74处可以把39份莲藕样品分为2大类，在0.80处可以分为5大类。表明莲藕的遗传差异较小，与传统的分类结果相似，为莲藕分类鉴定和优良品种选育作参考。 关键词：莲藕； DNA提取；正交设计；ISSR分子标记；遗传多样性 Analysis of genetic diversity of 39 Lotus root cultivars by using ISSR markers Abstract: This research optimized the DNA’s extraction method of lotus root. Used the orthogonal design scheme to optimize lotus’ ISSR - polymerase chain reaction (PCR) system from four factors and three levels, then used the optimized reaction system to screen primers and amplified the 39 samples of lotus’DNA with the seleted primers. The genetic diversity of the lotus were analysed. In the research, We obtained the best reaction system of lotus’ PCR amplification through the orthogonal experiment, using this system pick out eight of those lotus ISSR-PCR amplification primers from 18 designed primers. obtained 59 DNA bands totally, and there are 37 polymorphic bands, polymorphism ratio is 62.71%, every primers can amplify 7.36 bands on average. Using the NTSYS-pc software to analyse the lotus’ genetic diversity, result shows that genetic coefficient of 39 samples of lotus are between 0.63 and 0.98, use the UPGMA for clustering analysis, we can divide the 39 lotus samples into two types at the 0.74, and in 0.8 place they can be divided into 5 types. This papper provides the reference for identification , classification and fine varieties breeding of lotus. Key words: Lotus root; germplasm resources; extraction of DNA; ISSR; genetic diversity 目 录 文献综述………………………………………………………………………………1 1 莲藕的生物学特性……………………………………