發(fā)布時(shí)間：2018-07-02 10:22

  本文選題：白菜型油菜 + 甘藍(lán)�。� 參考：《西南大學(xué)》2017年碩士論文

【摘要】：油菜是世界范圍內(nèi)重要的油料作物和經(jīng)濟(jì)作物,但由腐生真菌核盤菌引起的油菜菌核病嚴(yán)重影響油菜的產(chǎn)量和品質(zhì)。生產(chǎn)實(shí)踐表明,選擇和培育抗(耐)病品種才是控制油菜菌核病最經(jīng)濟(jì)、長久的途徑。但油菜中并未發(fā)現(xiàn)高抗或者免疫的材料,這嚴(yán)重限制了油菜抗菌核病育種,所以研究者嘗試在近緣物種中挖掘優(yōu)良抗性來改良油菜菌核病抗性育種。在前期研究中,本實(shí)驗(yàn)室利用前期發(fā)現(xiàn)的高抗野生甘藍(lán)“C01”和感病栽培甘藍(lán)“C41”構(gòu)建的F2群體,定位出了5個菌核病抗性相關(guān)的QTLs,分別位于C1染色體上的q LR10-1和q LR10-7,C3染色體上的q LR10-8,以及C9染色體上的q SR09-1和q SR09-2。前期利用“C01”和白菜型油菜“6Y733”單倍體途徑,并以“6Y733”為輪回親本,已成功將C9染色體上的QTLs轉(zhuǎn)到“6Y733”中,獲得了莖稈抗性相比于親本“6Y733”提高1.7倍的白菜型油菜單株。在本研究中,欲將C1和C3染色體上的QTLs,通過分子輔助選擇聚合到白菜型油菜中,具體研究結(jié)果如下:1菌核病相關(guān)特異分子標(biāo)記的開發(fā)利用C1染色體上菌核病QTL區(qū)間連鎖的標(biāo)記序列,在甘藍(lán)參考基因組上比對獲得了一個1.56Mb的物理區(qū)段。根據(jù)抗病甘藍(lán)“C01”和感病甘藍(lán)“C41”重測序數(shù)據(jù),選擇兩材料間插入缺失大于20bp的位置開發(fā)了40個In Del標(biāo)記。利用“C01”、“C41”和“6Y733”篩選后獲得了9對多態(tài)性良好的標(biāo)記,最終有一個標(biāo)記成功添加到了C1染色體連鎖群中。2分子輔助選擇聚合抗性對本研究中聚合對象,首先利用C9染色體QTLs連鎖標(biāo)記進(jìn)行篩選,保證要聚合材料已含有C9染色體上QTLs。然后在要聚合的白菜型油菜后代中隨機(jī)選出6個材料,同抗病甘藍(lán)“C01”和“6Y733”兩親本一起,對C1和C3染色體上QTLs連鎖的SSR標(biāo)記及新開發(fā)的In Del標(biāo)記進(jìn)行多態(tài)性篩選,最后篩選出了3個多態(tài)性很好,且條帶清晰的標(biāo)記作為后續(xù)聚合輔助篩選標(biāo)記。經(jīng)過兩年分子輔助選擇的聚合育種,我們在含有C9抗病QTLs的群體中成功聚合了C1和C3染色體上的QTLs。對每年分子輔助選擇獲得的含C9外的QTLs連鎖標(biāo)記的材料(命名為NewQTL group)和不含任何QTL連鎖標(biāo)記的材料(命名為Non-QTL group)及親本進(jìn)行莖稈離體鑒定。第一年通過莖稈鑒定發(fā)現(xiàn),New-QTL group和Non-QTL group之間達(dá)到了極顯著差異(P0.0001),其莖桿抗性平均值分別為0.43±0.10和0.68±0.09。在New-QTL group中我們發(fā)現(xiàn)了三份莖稈抗性很好的材料,抗性值分別為0.34±0.032、0.37±0.059和0.32±0.119,相對于白菜親本莖稈抗性提高了約3倍。通過差異分析發(fā)現(xiàn)它們均分別相對兩親本達(dá)到了極顯著性差異(P0.0001)。我們將三份材料分別自交和雜交,對后代進(jìn)行第二年分子輔助選擇后,將New-QTL group和Non-QTL group材料進(jìn)行莖稈抗性鑒定,同樣發(fā)現(xiàn)兩群體間達(dá)到了極顯著差異(P0.0001),其莖桿抗性平均值分別為0.55±0.181和0.90±0.132,并最終在New-QTL group中發(fā)現(xiàn)了兩份莖稈抗性相對于白菜親本提高4倍多的材料,其抗性值分別為0.24±0.016和0.25±0.024,成功將C1和C3染色體上的QTLs聚合到了已含有C9染色體上的抗病QTL的白菜后代中。
[Abstract]:Rape is an important oil crop and economic crop in the world, but Sclerotinia sclerotiorum caused by Sclerotinia sclerotiorum seriously affects the yield and quality of rape. Production practice shows that selection and cultivation of resistant varieties is the most economical and long way to control rape sclerotinia, but there is no high resistance or immunity in rape. Material, which seriously restricts the breeding of rapeseed antibacterial nuclear disease, so researchers have tried to improve the resistance breeding of Brassica sclerotiorum in the near marginal species. In the earlier study, 5 Sclerotinia Sclerotinia were located with the high resistance wild cabbage "C01" and the susceptible cultivated Brassica napus "C41" in the earlier study. The sex related QTLs, Q LR10-1 and Q LR10-7 on the C1 chromosome, Q LR10-8 on the C3 chromosome, and Q SR09-1 on the C9 chromosome, and the use of the haploid pathway in the early stage of the C9 chromosome and the cabbage type rapeseed haploid pathway. The stem resistance of the stem was 1.7 times higher than that of the parent "6Y733". In this study, the QTLs on the C1 and C3 chromosomes was intended to be polymerized into Brassica napus by molecular assisted selection. The specific results were as follows: 1 the development of specific markers related to sclerotia related to the interval linkage of the QTL Sclerotinia on the C1 chromosome. Sequence, the physical section of a 1.56Mb was obtained by comparison of the cabbage reference genome. 40 In Del markers were developed based on the disease resistant cabbage "C01" and the "C41" re sequencing data of the susceptible Brassica oleracea. The insertion loss between two materials was greater than that of 20bp. 9 pairs of polymorphisms were obtained with "C01", "C41" and "6Y733" screening. In the end, a marker was successfully added to the C1 chromosome chain to select the.2 molecule to assist the polymerization resistance in this study. First, the C9 chromosome QTLs linkage marker was used to ensure that the polymeric material contained QTLs. on the C9 chromosome and then selected 6 materials randomly in the descendants of the aggregate Brassica napus. Together with the two parents of Brassica oleracea "C01" and "6Y733", the QTLs linked SSR markers on the C1 and C3 chromosomes and the newly developed In Del markers were selected for polymorphism screening. Finally, 3 good polymorphic markers were selected as a follow-up polymerization auxiliary screening marker. After two years of molecular assisted selection, we were selected for the polymerization breeding. In the population containing the C9 resistant QTLs, the C1 and C3 chromosomes were successfully polymerized by QTLs. for each year molecular assisted selection of QTLs linked markers (named NewQTL group) and non QTL linked markers (named Non-QTL group) and parental identification in vitro. The first year was identified by stem identification. There were significant differences between New-QTL group and Non-QTL group (P0.0001). The average stem resistance of the stem was 0.43 + 0.10 and 0.68 + 0.09. in New-QTL group. We found three stalk resistant materials, and the resistance values were 0.34 + 0.032,0.37 + 0.059 and 0.32 + 0.119 respectively. 3 times. The difference was found to be significantly different from two parents (P0.0001). The three materials were interbred and hybridized respectively. After second years of molecular assisted selection, the resistance of New-QTL group and Non-QTL group was identified, and the significant difference was found among the two groups (P0 .0001), the average stem resistance of the stem was 0.55 + 0.181 and 0.90 + 0.132 respectively, and in the final New-QTL group, two stem resistance was found to be more than 4 times more than that of the cabbage parent. The resistance values were 0.24 + 0.016 and 0.25 + 0.024 respectively, and the QTLs on the C1 and C3 chromosomes was successfully polymerized to the disease resistant QTL containing the C9 chromosome. In the offspring of Chinese cabbage.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：Q943;S435.654

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