花生栽培种遗传多样性的SSR分析及其指纹图谱的构建

发布时间：2018-01-20 13:26

本文关键词： 花生 SSR 荧光检测法遗传多样性指纹图谱　出处：《曲阜师范大学》2013年硕士论文　论文类型：学位论文

【摘要】：花生是重要的油料作物和经济作物。中国拥有较为丰富的种质资源，但是随着针对性育种工作的广泛进行，花生栽培种遗传背景越来越狭窄。遗传多样性研究是进行花生栽培种育种改良的基础，而品种的指纹图谱鉴定可为育种工作的知识产权保护提供科学依据。本文以筛选出的100对SSR引物分析144份花生栽培种的遗传多样性，并对银染和荧光检测技术两种SSR PCR扩增产物的检测方法进行对比，同时选取23对引物的159个标记构建群体的指纹图谱，进而为育种提供分子依据、为生产提供服务。主要研究结果如下： 1.144份花生栽培种的遗传多样性分析：应用100对SSR引物对144个花生栽培种进行PCR扩增，共检测到409个等位变异，每对SSR引物平均能够检测到4.09个等位变异；聚类分析结果表明，100对SSR引物能够将供试的所有品种区分开，聚类结果与栽培花生类型、产地及亲缘关系呈正相关；品种遗传相似系数为0.68-0.97。可见，总体上栽培花生遗传基础较为狭窄，特别是国内现有的育成品种。多样性分析显示，地方品种和国外品种与其他品种间遗传距离较大，作为亲本应用于育种可拓宽选育品种的遗传背景。 2.银染和荧光检测技术的对比：选取9对多态性及特异性较好的引物，分别用银染法和荧光检测法扫描了85个栽培品种的SSR标记多态性。与银染法相比，，荧光检测法具有灵敏度高、准确性强、效率高、在遗传多样性分析方面更精确可靠等优点。此外，通过高通量PCR、高通量上样检测及大片段检测优化等方法可以提高荧光检测法检测效率，有效降低实验成本。 3.144份花生栽培种SSR指纹图谱的构建：应用23对等位变异数目大于5的SSR引物构建了144个供试材料的指纹图谱，以0、1形式统计，并将0、1字符串转换为十六进制，结果表明每一个品种的指纹图谱均为唯一代码。下一步将通过F1群体或亲缘关系较近的品种对构建的指纹图谱进行验证，以期为花生育种提供科学准确的鉴定手段。
[Abstract]:Peanut is an important oil crop and cash crop. China has abundant germplasm resources, but with the extensive work of targeted breeding. The genetic diversity of peanut cultivars is becoming more and more narrow, which is the basis of peanut breeding. The identification of fingerprinting can provide a scientific basis for the protection of intellectual property rights in breeding. The genetic diversity of 144 peanut cultivars was analyzed with 100 pairs of SSR primers selected in this paper. Silver staining and fluorescence detection were compared between two SSR PCR amplification methods, and 159 markers of 23 pairs of primers were selected to construct the fingerprint of the population. The main results are as follows: Analysis of genetic diversity of 1.144 peanut cultivars: #number0# pairs of SSR primers were used to amplify 144 peanut cultivars and 409 alleles were detected. An average of 4.09 alleles per pair of SSR primers could be detected. The results of cluster analysis showed that all cultivars could be distinguished by SSR primer, and the cluster results were positively correlated with the type, origin and relatives of cultivated peanut. The genetic similarity coefficient of cultivars was 0.68-0.97.The genetic basis of cultivated peanut was relatively narrow in general, especially the existing varieties in China. The diversity analysis showed. The genetic distance between local varieties and foreign varieties and other varieties is great. As parents, the genetic background of selected varieties can be widened. 2. Comparison of silver staining and fluorescence detection: 9 pairs of primers with good polymorphism and specificity were selected. The polymorphism of SSR markers in 85 cultivars were scanned by silver staining method and fluorescence detection method respectively. Compared with silver staining method, fluorescent detection method has higher sensitivity, higher accuracy and higher efficiency. In addition, the efficiency of fluorescence detection can be improved by high-throughput PCR, high-throughput sample detection and optimization of large fragment detection. Effectively reduce the cost of the experiment. Construction of SSR fingerprinting of 3. 144 peanut cultivars: using 23 pairs of SSR primers with allelic variation of more than 5, the fingerprints of 144 tested materials were constructed, and the results were analyzed in the form of 0 ~ 1. The result shows that the fingerprint of each variety is the unique code. The next step is to verify the fingerprint map by F1 population or the closely related variety. In order to provide a scientific and accurate identification means for peanut breeding.
【学位授予单位】：曲阜师范大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：S565.2

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