摘要石斛是兰科石斛属植物的统称，不仅有观赏性可作为观赏植物而且有较高的药用价值可用于生产铁皮枫斗等药品。在本课题中我们优化了石斛属植物DNA提取方法，获得了浓度高、杂质少的石斛基因组DNA，优化了聚丙烯酰胺凝胶电泳技术以及银染方法，使其操作更加简便、快捷。从367对可能具有特异性的SSR分子标记引物中，本课题成功的筛选出57对具有特异性的SSR分子标记引物，这些特异性引物可用于不同石斛属植物种质资源的遗传多样性分析以及杂交群体的鉴定。本课题组总共收集了50种不同石斛属植物材料，通过SSR分子标记技术，我们对这些不同石斛属植物材料的遗传多样性进行分析和研究。主要实验过程：通过对不同石斛属植物材料基因组DNA的提取，我们优化了实验步骤，获得了浓度高、杂质少的石斛属植物材料的基因组DNA；通过SSR-PCR扩增技术，我们优化了扩增体系，得到了含有目的DNA片段的扩增产物；利用聚丙烯酰胺凝胶电泳以及银染技术，我们缩短了银染的时间，简化了实验步骤，得到了条带清晰的电泳图；通过对扩增产物中目的DNA片段的回收，我们获得了浓度高、纯度好的目的DNA溶液；通过TA克隆，将目的DNA片段与T载体相连接，把含有目的DNA片段的质粒转化到感受态细胞中去，在筛选培养基上获得单克隆菌落，摇菌后将加有甘油的菌液送至公司测序，我们得到了目的DNA片段的核苷酸序列。利用MEGA生物学软件构建了不同石斛属植物材料的系统进化树，通过数据分析，我们将其中37种不同石斛属材料划分为四大类，可以判断不同石斛属植物的亲缘关系，为不同石斛属植物的杂交育种提供理论指导。本课题研究成果可以提高药用石斛植物的产量，提高经济收入；保护濒临灭绝的珍惜石斛属植物品种种质资源；还可以用于不同石斛属植物的品种鉴定，与传统方法相比，能够更加准确、迅速的鉴定出不同石斛品种。88478

毕业论文关键词：石斛； 种质资源； SSR标记； 遗传多样性

Abstract Dendrobium is the genus Dendrobium orchid collectively, not only ornamental can be used as ornamental plants and have high medicinal value can be used for the production of metal powder and other drugs。 In this paper, we optimized the DNA extraction method of Dendrobium, obtained the genomic DNA of Dendrobium with high concentration and low impurity, optimized the polyacrylamide gel electrophoresis and silver staining method, and made it easier and faster to operate。From the 367 pairs of SSR molecμLar marker primers which may have specificity, 57 pairs of SSR molecμLar marker primers with specificity can be successfμLly screened out。 These specific primers can be used for the genetic persity analysis of different Dendrobium germplasm resources and Identification of hybrid popμLations。A total of 50 different Dendrobium plant materials were collected。 Through SSR molecμLar marker technique, we analyzed and studied the genetic persity of these different Dendrobium plant materials。 The main experimental process: By extracting the genomic DNA of different Dendrobium plant materials, we optimized the experimental steps and obtained the genomic DNA of Dendrobium plant material with high concentration and low impurity。 Through SSR-PCR, we optimized the We used to shorten the time of silver staining and simplify the experimental steps, and get a clear electrophoregram of the band。 By using polyacrylamide gel electrophoresis and silver staining technology, The target DNA fragment was ligated with the T vector by TA cloning, and the plasmid containing the target DNA fragment was transformed into the competent cells。 The DNA fragment containing the target DNA fragment was transformed into the competent DNA fragment by TA cloning。 To obtain monoclonal colonies on the screening medium, the bacterial solution with glycerol was sent to the company for sequencing。 We obtained the nucleotide sequence of the target DNA fragment。The phylogenetic tree of Dendrobium plant was constructed by MEGA biology software。 Through data analysis, we pided 37 different Dendrobium species into four categories, which coμLd determine the genetic relationship of different Dendrobium species, Plant breeding to provide theoretical guidance。 The research resμLts of this project can improve the yield of medicinal Dendrobium plants and improve the economic income; protect the endangered species of Dendrobium species germplasm resources; can also be used for different species of Dendrobium species identification, compared with the traditional method can be more Accurate and rapid identification of different varieties of Dendrobium。源Y于Y优E尔Y论L文W网wwW.yOueRw.com 原文+QQ752018.766 基于SSR序列的石斛遗传多样性评价:http://www.youerw.com/shengwu/lunwen_161438.html