水稻抗白叶枯病品种H120-2-1的遗传分析及其基因定位

水稻抗白叶枯病品种H120-2-1的遗传分析及其基因定位(8400字)

摘  要：H120-2-1是高感品种丽江新团黑谷通过航天诱变获得抗白叶枯病材料，对广东省优势菌系Ⅴ型菌表现高水平抗病性。为了有效地利用该基因所控制的抗性，对H120-2-1与籼稻感病品种IR24的杂交组合由来的998个F2个体，接种对双亲品种表现分明的非亲和性/亲和性反应的菌株SCB4-1，结果表明该F2 群体中抗病植株与感病植株的分离比符合3：1（750R：248S），由此推断H120-2-1所表现的对接种菌株SCB4-1的抗性是由一对显性基因控制的。为了快速确定主效抗性基因的染色体位置，构建了由25个抗病个体和248个感病个体组成的F2代作图群体，并利用微卫星分子标记技术，结合基于分离群体分析法 (bulked-segregant analysis, BSA) 的隐性群体分析法 (reccesive-class analysis, RCA) 对该作图群体进行了连锁分析。首先，根据分离群体分析法分别用25个抗病和感病个体的DNA构建了抗病基因池和感病基因池。然后，利用从水稻12条染色体上选出的250个微卫星标记对两个亲本和抗、感基因池进行了筛选，获得了两个位于第11染色体上的候选SSR标记RM206和RM457。为了进一步确定这两个候选标记，利用隐性群体分析法对248个感病个体进行了连锁分析，结果表明它们与目的基因连锁，重组率分别为5.7% 和27.9%。本文因此将目的基因暂时命名为Xa-H120-2-1(t)。本研究为抗性基因Xa-H120-2-1(t)的精细定位、分子标记辅助育种奠定了坚实的基础。

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关键词：水稻白叶枯病；抗性基因；微卫星；连锁分析

Bacterial blight resistance varieties H120-2-1 genetic analysis and gene mapping

Abstract：H120-2-1 is highly susceptible Lijiangxintuanheigu Valley obtained by space mutagenesis bacterial blight resistance materials, the dominant strains in Guangdong Province Ⅴ type strain show a high level resistance. In order to effectively control the use of the resistance gene on the H120-2-1 and the susceptible cultivar IR24 indica hybrid combination of the origin of the 998 F2 individuals, vaccination of the parent species show distinct non-affinity / affinity reaction strains SCB4-1, results show that the F2 populations resistant plants and susceptible plants in the segregation ratio consistent with 3:1 (750R: 248S), infer H120-2-1 shown on the inoculated SCB4-1 resistance by a dominant gene control. In order to quickly identify major resistance genes in the chromosomal location, build resistance to disease by 25 to 248 individuals and susceptible individuals in the F2 generation of mapping population composition, and using microsatellite markers, combined with population analysis based on separation (bulked -segregant analysis, BSA) in the recessive group analysis (reccesive-class analysis, RCA) mapping population was the linkage analysis. First, according to analysis were used to separate groups of 25 individual resistant and susceptible DNA pool of resistance genes was constructed and susceptible gene pool. Then, on the 12 rice chromosomes from the selected 250 microsatellite markers on both the parents and resistant and susceptible gene pool were screened, won two in a candidate on chromosome 11 SSR markers RM206 and RM457. To further mark the two candidates, the use of implicit group analysis of 248 individuals were susceptible linkage analysis, the results show that they are linked with the target gene, recombination rates were 5.7% and 27.9%. This paper will therefore be the target gene, tentatively named Xa-H120-2-1 (t). This study resistance gene Xa-H120-2-1 (t) the fine mapping, marker assisted breeding and laid a solid foundation.

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    Key words: Bacterial blight; resistance gene; microsatellite; linkage analysis

目  录

摘  要    1
关键词    1
1 前 言    2
2材料    3
2.1 实验材料及设备    3
2.1.1 水稻材料    3
2.1.2 水稻白叶枯病菌株    3
2.1.3 主要试剂    3
2.1.4 主要仪器设备    3
2.2 试验方法    4
2.2.1 作图群体构建及表型分析    4
2.2.2 植物DNA提取    4
2.2.3  SSR分析    5
2.2.4 电泳检测    6
2.2.5 抗病基因池和感病基因池的构建    6
2.2.6 标记筛选及连锁分析    7
2.2.7 数据分析    7
3 结果与分析    7
3.1 目标基因分子定位    7
3.1.1 作图群体的构建    7
3.1.2 植物DNA提取及质量检测    8
3.1.3抗性基因的定位    8
4 讨论与结论    10
4.1 影响精细定位的因素    10
4.1.1 作图群体的选择    10
4.1.2 菌系的选择    11
参考文献    11
致    谢    12 [资料来源：http://Doc163.com]