甘蓝型油菜遗传多样性分析及核心亲本的指纹图谱构建

发布时间：2018-06-04 08:15

本文选题：甘蓝型油菜 + SSR分子标记　；参考：《华中农业大学》2013年硕士论文

【摘要】：我国是世界上最大的油菜生产国,也是世界上第一个成功大面积利用油菜杂种优势的国家。油菜杂交种由于具有产量高、抗逆性强和适应性广等优点,深受种子企业的亲睐和广大农民欢迎。但随着市场商业化的不断推进,不法经营者的人为掺假或以次充好以及混杂或以不正当手段获取育种家的亲本用于生产杂交种,损害了农民、企业和育种家的合法利益。因此,迫切需要建立对杂交油菜品种及其亲本进行真实性和纯度鉴定的技术体系,为亲本材料的知识产权保护和维护农民的合法权益提供技术支撑。 SSR标记由于具有共显性、稳定性好、遗传方式简单以及能检测多个等位位点等优点,被广泛应用于玉米、水稻等主要农作物品种的纯度和真实性鉴定。本研究利用SSR标记对本实验室的120份甘蓝型油菜种质资源进行分析,筛选出一批多态性丰富的SSR引物,同时构建了骨干亲本SSR-DNA的指纹图谱,为油菜杂交种纯度鉴定及其亲本的真实性提供了可靠的参考依据。主要结果如下： 1、从120份材料中选取6个不同遗传背景的材料所组成小样本(包括2个不育系、2个恢复系和2个常规品系),对本实验室拥有的1938对SSR引物进行筛选,共得到多态性引物543对,经过比较选择出带型清晰、稳定的SSR引物173对。 2、用173对引物对120份甘蓝型油菜材料进行分析,共扩增出646条带,其中多态性条带512条,多态性比率为80%。一对SSR引物扩增多态性带数为2-8个,平均为3.8个；信息多态量PIC值变化范围0.05-0.92,平均为0.61。 3、利用512条多态性条带对120份材料进行UMPGMA聚类分析,以遗传相似系数0.61为阀值,将120份材料分为A、B两个大类。A类共100份材料,在相似系数0.69处划分4个亚类,其中大部分不育系材料聚在第一亚类和第二亚类,说明不育系材料遗传变异相对较小；而42份恢复系在两大类中均有分布,表明恢复系材料遗传变异比较丰富。 4、根据173对引物在其中32份核心亲本中的扩增结果,对有条带出现赋值为“1”,无条带出现赋值为“0”,构建了32份甘蓝型油菜核心亲本的标准数字化指纹图谱。同时初步设计亲本材料图谱快速查询计算机数据库程序。
[Abstract]:China is the largest rapeseed producer in the world and the first country in the world to successfully utilize heterosis in large area. Rapeseed hybrids are popular among seed enterprises and farmers because of their high yield, strong resistance and wide adaptability. But with the development of commercialization of the market, the illegal operators' artificial adulteration or subdued, mixed or improper means to obtain the parents of breeders is used to produce hybrids, which is harmful to the legitimate interests of farmers, enterprises and breeders. Therefore, it is urgent to establish a technical system to identify the authenticity and purity of hybrid rape varieties and their parents so as to provide technical support for protecting the intellectual property rights of parents and safeguarding the legitimate rights and interests of farmers. Because of its advantages of co-dominance, good stability, simple genetic mode and the ability to detect multiple alleles, SSR markers have been widely used to identify the purity and authenticity of major crop varieties such as maize and rice. In this study, 120 Brassica napus germplasm resources in our laboratory were analyzed by SSR markers, and a batch of polymorphic SSR primers were screened, and the fingerprints of the backbone parent SSR-DNA were constructed. It provides a reliable reference for the purity identification of rapeseed hybrids and the authenticity of their parents. The main results are as follows: 1. A total of 1938 pairs of SSR primers were screened from 1938 pairs of polymorphic primers (including 2 male sterile lines, 2 restorer lines and 2 conventional lines) from 6 materials of different genetic backgrounds, and a total of 543 pairs of polymorphic primers were obtained. After comparison, 173 pairs of SSR primers with clear band and stable pattern were selected. 2, 120 rape (Brassica napus L.) materials were analyzed with 173 pairs of primers, and 646 bands were amplified, of which 512 were polymorphic bands, with a polymorphism ratio of 80000. The number of polymorphic bands amplified by a pair of SSR primers ranged from 2 to 8, with an average of 3.8, and the range of PIC values of information polymorphism was 0.05-0.92, with an average of 0.61. 3. Using 512-polymorphic bands, 120 materials were analyzed by UMPGMA cluster analysis. The genetic similarity coefficient was 0.61 as the threshold value, and 120 materials were divided into two categories. A group of 100 materials were divided into four subgroups at the similarity coefficient of 0.69. Most of the male sterile lines were clustered in the first and second subclasses, indicating that the genetic variation of the male sterile lines was relatively small, while 42 restorer lines were distributed in the two major categories, indicating that the restorer lines had relatively rich genetic variation. 4. According to the amplification results of 173 pairs of primers in 32 core parents, the standard digital fingerprint of 32 core parents of Brassica napus was constructed. At the same time, the computer database program of parent material atlas was designed.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：S565.4

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