本文關(guān)鍵詞： 甘藍 雜種致死 Bateson-Dobzhansky-Muller模式 精細定位　出處：《中國農(nóng)業(yè)科學(xué)院》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：雜種致死是一種合子后的生殖隔離類型,具體表現(xiàn)為雙親可以正常的生長,但其F1雜交種在性成熟前出現(xiàn)死亡的現(xiàn)象。對雜種致死進行基因定位是克隆的分子基礎(chǔ),為闡明雜種致死的作用機理奠定基礎(chǔ)。本研究中,我們發(fā)現(xiàn)了甘藍自交系09-211和09-222的F1出現(xiàn)雜種致死現(xiàn)象。通過對甘藍雜種致死的遺傳分析,發(fā)現(xiàn)雜種致死是由兩個顯性互補的核基因BoHL1和BoHL2控制,分別來自09-211和09-222,其遺傳模式符合雙位點的Bateson-Dobzhansky-Muller(BDM)模式。在此基礎(chǔ)上,本研究利用攜帶致死基因BoHL2的材料09-222和不攜帶雜種致死基因的材料87-534、11-196和96-100進行雜交獲得F1,利用獲得的F1代雜種和攜帶致死基因BoHL1的材料09-211進行二次雜交獲得分離群體,對甘藍雜種致死基因BoHL2進行精細定位,為甘藍雜種致死基因的克隆和功能分析奠定基礎(chǔ)。研究結(jié)果如下:1.甘藍雜種致死基因BoHL2的初定位通過致死分離群體1:(09-222×87-534)×09-211共計336株,利用BSA法在C04染色體上找到6對和BoHL2連鎖的標記:D349、InDel0110、InDel0112、InDel0114、LTSSR342和LTSSR346,連鎖分析發(fā)現(xiàn)BoHL2位于LTSSR346和InDel0114之間,物理距離6.65 Mb。通過對含致死基因的雙親09-211和09-222進行10×重測序,在C04染色體LTSSR346和InDel0114之間6.65 Mb區(qū)間內(nèi)設(shè)計開發(fā)了73對Indel引物,找到33對多態(tài)性好的引物。擴大分離群體1將BoHL2初步定位到HL211和HL249之間,物理距離1.8 Mb。2.甘藍雜種致死基因BoHL2的精細定位利用分離群體(09-222×11-196)×09-211和(09-222×96-100)×09-211將BoHL2精細定位到C04染色體的HL235和HL234之間,兩個標記和BoHL2的遺傳距離分別是0.3 cM和0.3cM,物理距離是70.33 Kb。3.候選基因的分析根據(jù)甘藍基因組注釋信息,發(fā)現(xiàn)定位區(qū)間內(nèi)共含9個基因,其中Bol033387和Bol033390具有抗病基因的結(jié)構(gòu)。通過對5份含雜種致死基因的甘藍自交系的定位區(qū)間內(nèi)的Indel和SNP分析發(fā)現(xiàn)變異位點集中在Bol033385基因,預(yù)測這三個基因為候選基因。
[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.
【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S635

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本文編號：1506830