甘蓝杂种致死基因BoHL2精细定位的研究

发布时间：2018-02-12 23:40

本文关键词： 甘蓝杂种致死 Bateson-Dobzhansky-Muller模式精细定位　出处：《中国农业科学院》2016年硕士论文　论文类型：学位论文

【摘要】：杂种致死是一种合子后的生殖隔离类型,具体表现为双亲可以正常的生长,但其F1杂交种在性成熟前出现死亡的现象。对杂种致死进行基因定位是克隆的分子基础,为阐明杂种致死的作用机理奠定基础。本研究中,我们发现了甘蓝自交系09-211和09-222的F1出现杂种致死现象。通过对甘蓝杂种致死的遗传分析,发现杂种致死是由两个显性互补的核基因BoHL1和BoHL2控制,分别来自09-211和09-222,其遗传模式符合双位点的Bateson-Dobzhansky-Muller(BDM)模式。在此基础上,本研究利用携带致死基因BoHL2的材料09-222和不携带杂种致死基因的材料87-534、11-196和96-100进行杂交获得F1,利用获得的F1代杂种和携带致死基因BoHL1的材料09-211进行二次杂交获得分离群体,对甘蓝杂种致死基因BoHL2进行精细定位,为甘蓝杂种致死基因的克隆和功能分析奠定基础。研究结果如下:1.甘蓝杂种致死基因BoHL2的初定位通过致死分离群体1:(09-222×87-534)×09-211共计336株,利用BSA法在C04染色体上找到6对和BoHL2连锁的标记:D349、InDel0110、InDel0112、InDel0114、LTSSR342和LTSSR346,连锁分析发现BoHL2位于LTSSR346和InDel0114之间,物理距离6.65 Mb。通过对含致死基因的双亲09-211和09-222进行10×重测序,在C04染色体LTSSR346和InDel0114之间6.65 Mb区间内设计开发了73对Indel引物,找到33对多态性好的引物。扩大分离群体1将BoHL2初步定位到HL211和HL249之间,物理距离1.8 Mb。2.甘蓝杂种致死基因BoHL2的精细定位利用分离群体(09-222×11-196)×09-211和(09-222×96-100)×09-211将BoHL2精细定位到C04染色体的HL235和HL234之间,两个标记和BoHL2的遗传距离分别是0.3 cM和0.3cM,物理距离是70.33 Kb。3.候选基因的分析根据甘蓝基因组注释信息,发现定位区间内共含9个基因,其中Bol033387和Bol033390具有抗病基因的结构。通过对5份含杂种致死基因的甘蓝自交系的定位区间内的Indel和SNP分析发现变异位点集中在Bol033385基因,预测这三个基因为候选基因。
[Abstract]:The death of hybrids is a type of reproductive isolation after zygote, which shows that both parents can grow normally, but the F1 hybrids die before sexual maturation. Gene mapping of hybrid death is the molecular basis of cloning. In order to elucidate the mechanism of hybrid death, we found that F _ 1 of the inbred lines 09-211 and 09-222 of Brassica oleracea had the phenomenon of hybrid death. The genetic analysis of hybrid death in Brassica oleracea was carried out. It was found that hybrid death was controlled by two dominant complementary nuclear genes, BoHL1 and BoHL2, respectively, from 09-211 and 09-222, respectively. The genetic model was in accordance with the Bateson-Dobzhky-Mullerger BDM pattern. In this study, F _ 1 was obtained by hybridization with the material 09-222, which carried the lethal gene BoHL2, and the material 87-534n11-196 and 96-100, which did not carry the hybrid lethal gene, and the F1 hybrid and the material carrying the lethal gene BoHL1, 09-211, were used for secondary hybridization to obtain the isolated population. The fine mapping of the hybrid lethal gene BoHL2 was carried out, which laid a foundation for the cloning and functional analysis of the hybrid lethal gene of cabbage. The results are as follows: 1. The initial location of the hybrid lethal gene BoHL2 of cabbage was obtained by 1: 09-222 脳 87-534 脳 09-211, a total of 336 strains. By using BSA method, 6 pairs of markers linked to BoHL2 were found on chromosome C04:: D349 / InDel0110 / InDel0112 / InDel0114 / LTSSR342 and LTSSR346. linkage analysis showed that BoHL2 was located between LTSSR346 and InDel0114 with a physical distance of 6.65 Mb. 10 脳 sequencing was carried out on parents 09-211 and 09-222 containing lethal genes. 73 pairs of Indel primers were designed and developed in the 6.65Mb interval between C04 chromosome LTSSR346 and InDel0114. 33 pairs of polymorphic primers were found. Physical distance 1.8 Mb 路2.The fine mapping of the lethal gene BoHL2 in Brassica oleracea by using the isolated populations of 09-222 脳 11-196) 脳 09-211 and 09-222 脳 96-100) 脳 09-211 mapped BoHL2 between HL235 and HL234 on chromosome C04. The genetic distances of the two markers and BoHL2 were 0.3cM and 0.3cM, respectively. The physical distance was 70.33Kb.3.According to the genome annotation information of cabbage, it was found that there were nine genes in the locus. Among them, Bol033387 and Bol033390 had the structure of disease resistance genes. By analyzing the Indel and SNP of five inbred lines containing hybrid lethal genes, it was found that the mutation sites were concentrated in Bol033385 genes, and the candidate genes were predicted.
【学位授予单位】：中国农业科学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S635

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