大豆细胞质雄性不育系SXCMS1A的育性恢复性遗传和恢复基因SSR标记定位

发布时间：2018-12-20 16:10

【摘要】：大豆杂种优势利用是目前大豆研究的一个重要领域,也是提高大豆单产、挖掘大豆生产潜力的一条有效途径。目前我国大豆杂种优势利用研究已取得了阶段性的成果,杂交大豆的高效繁制种技术,强优势组合的选育,杂交种的培育,均已取得长足的发展。然而要实现杂交大豆产业化仍然需要进一步提高杂种优势利用的潜力,需要扩大不育系和恢复系的选育范围、加快选育速度,这样势必要求在细胞质雄性不育系的遗传模式和分子标记辅助选择等基础领域中继续深入探究。本研究中利用大豆细胞质雄性不育系SXCMS1A与恢复系AHH-8进行杂交,构建了F1杂交群体和F2育性分离群体,通过花粉镜检结合成熟期植株育性表现来判定单株育性,获得了植株育性分离情况。根据F1、F2群体育性分离情况讨论了不育基因同恢复基因的数量及显隐性关系,明确了大豆细胞质雄性不育系SXCMS1A的遗传模式;利用445对大豆SSR引物在亲本和基因池间进行多态性筛选,将有多态性的引物在F2分离群体中进行了单株验证,经SSR结果过分析对恢复基因进行了初步定位。获得了以下结果:1.以大豆细胞质雄性不育系SXCMS1A为母本,以恢复系AHH-8为父本进行杂交,获得了F1杂交群体和F2分离群体。通过对植株花粉进行育性镜检,并结合成熟期田间育性调查判断F1杂交群体和F2分离群体的单株育性。结果F1群体的306株单株全部表现半不育,F2群体的293株单株中可育株、半不育株分别有139株、154株,经χ2适合性测验检测分离比符合1:1。根据细胞质雄性不育育性遗传规律推测大豆细胞质雄性不育系SXCMS1A是典型单基因控制的配子体不育。2.判定单株育性的实验中采用花粉败育率镜检和观察成熟期单株株型特征相结合的方法能够有效降低因采集花蕾数量不足造成的镜检误差,同时可以避免由于昆虫传粉某些不育株大量结荚造成的育性误判,进而影响实验的准确性。3.在遗传模式研究的基础上,选用445对大豆SSR引物在亲本和基因池之间进行多态性筛选,将筛选到的具有多态性的引物在F2分离群体中进行了单株验证。通过SSR标记分析,获得了位于D1a连锁群上的2个与恢复基因连锁的SSR标记Satt184和Satt267,与恢复基因的遗传距离分别为17.3cM和10.2cM。利用Join Map 4.0软件进行了遗传作图分析,将恢复基因定位在D1a连锁群上。
[Abstract]:The utilization of soybean heterosis is an important field of soybean research at present. It is also an effective way to improve soybean yield and tap soybean production potential. At present, the research on the utilization of soybean heterosis in China has achieved a lot of achievements. The high efficient breeding techniques, the breeding of strong heterosis combinations and the breeding of hybrids have made great progress in hybrid soybean breeding. However, in order to realize the industrialization of hybrid soybean, it is necessary to further improve the potential of utilization of heterosis, to expand the breeding range of sterile lines and restorer lines, and to speed up the breeding speed. Therefore, it is necessary to further explore the genetic model and molecular marker assisted selection of cytoplasmic male sterile lines. In this study, F1 hybrid population and F2 fertility segregation population were constructed by hybridization between soybean cytoplasmic male sterile line SXCMS1A and restorer line AHH-8, and single plant fertility was determined by pollen microscopy combined with fertility performance of mature plant. The isolation of plant fertility was obtained. According to the physical segregation of F _ 1 / F _ 2 group, the number and recessive relationship between sterile gene and restorer gene were discussed, and the genetic model of cytoplasmic male sterility line SXCMS1A in soybean was determined. Four hundred and forty-five pairs of soybean SSR primers were used to screen the polymorphism between parents and gene pools. The polymorphic primers were tested in F2 population. The restoring genes were preliminarily located by SSR analysis. The following results are obtained: 1. F1 hybrid population and F2 segregated population were obtained by crossing soybean cytoplasmic male sterile line SXCMS1A as female parent and restorer line AHH-8 as male parent. The fertility of F _ 1 hybrid population and F _ 2 segregated population was determined by microscopical examination of pollen fertility and field fertility investigation at maturity stage. Results there were 139 fertile plants and 154 semi-sterile plants in 293 single plants of F _ 2 population. The segregation ratio was 1: 1 by 蠂 ~ 2 fitness test. According to the genetic law of fertility of cytoplasmic male sterility, it is inferred that the cytoplasmic male sterile line SXCMS1A is a typical gametophyte sterility controlled by single gene. In the experiment of determining the fertility of a single plant, the method of pollen abortion rate and observation of the characteristics of individual plant type at maturity stage can effectively reduce the error caused by insufficient number of collected flower buds. At the same time, it can avoid the misjudgment of fertility caused by a large number of sterile plants of insect pollination, thus affecting the accuracy of the experiment. Based on the study of genetic pattern, a total of 445 pairs of soybean SSR primers were selected to screen polymorphism between parent and gene pool, and the polymorphic primers were tested on a single plant in F2 segregated population. By SSR marker analysis, the genetic distances between the two SSR markers Satt184 and Satt267, linked to the restorer gene were obtained to be 17.3cM and 10.2cM, respectively, which were located on the D1a linkage group. Genetic mapping was carried out with Join Map 4.0 software, and the restoration gene was located on the D1a linkage group.
【学位授予单位】：山西大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S565.1

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