利用野生甘蓝改良白菜型油菜菌核病抗性

发布时间：2018-07-02 10:22

本文选题：白菜型油菜 + 甘蓝　；参考：《西南大学》2017年硕士论文

【摘要】：油菜是世界范围内重要的油料作物和经济作物,但由腐生真菌核盘菌引起的油菜菌核病严重影响油菜的产量和品质。生产实践表明,选择和培育抗(耐)病品种才是控制油菜菌核病最经济、长久的途径。但油菜中并未发现高抗或者免疫的材料,这严重限制了油菜抗菌核病育种,所以研究者尝试在近缘物种中挖掘优良抗性来改良油菜菌核病抗性育种。在前期研究中,本实验室利用前期发现的高抗野生甘蓝“C01”和感病栽培甘蓝“C41”构建的F2群体,定位出了5个菌核病抗性相关的QTLs,分别位于C1染色体上的q LR10-1和q LR10-7,C3染色体上的q LR10-8,以及C9染色体上的q SR09-1和q SR09-2。前期利用“C01”和白菜型油菜“6Y733”单倍体途径,并以“6Y733”为轮回亲本,已成功将C9染色体上的QTLs转到“6Y733”中,获得了茎秆抗性相比于亲本“6Y733”提高1.7倍的白菜型油菜单株。在本研究中,欲将C1和C3染色体上的QTLs,通过分子辅助选择聚合到白菜型油菜中,具体研究结果如下:1菌核病相关特异分子标记的开发利用C1染色体上菌核病QTL区间连锁的标记序列,在甘蓝参考基因组上比对获得了一个1.56Mb的物理区段。根据抗病甘蓝“C01”和感病甘蓝“C41”重测序数据,选择两材料间插入缺失大于20bp的位置开发了40个In Del标记。利用“C01”、“C41”和“6Y733”筛选后获得了9对多态性良好的标记,最终有一个标记成功添加到了C1染色体连锁群中。2分子辅助选择聚合抗性对本研究中聚合对象,首先利用C9染色体QTLs连锁标记进行筛选,保证要聚合材料已含有C9染色体上QTLs。然后在要聚合的白菜型油菜后代中随机选出6个材料,同抗病甘蓝“C01”和“6Y733”两亲本一起,对C1和C3染色体上QTLs连锁的SSR标记及新开发的In Del标记进行多态性筛选,最后筛选出了3个多态性很好,且条带清晰的标记作为后续聚合辅助筛选标记。经过两年分子辅助选择的聚合育种,我们在含有C9抗病QTLs的群体中成功聚合了C1和C3染色体上的QTLs。对每年分子辅助选择获得的含C9外的QTLs连锁标记的材料(命名为NewQTL group)和不含任何QTL连锁标记的材料(命名为Non-QTL group)及亲本进行茎秆离体鉴定。第一年通过茎秆鉴定发现,New-QTL group和Non-QTL group之间达到了极显著差异(P0.0001),其茎杆抗性平均值分别为0.43±0.10和0.68±0.09。在New-QTL group中我们发现了三份茎秆抗性很好的材料,抗性值分别为0.34±0.032、0.37±0.059和0.32±0.119,相对于白菜亲本茎秆抗性提高了约3倍。通过差异分析发现它们均分别相对两亲本达到了极显著性差异(P0.0001)。我们将三份材料分别自交和杂交,对后代进行第二年分子辅助选择后,将New-QTL group和Non-QTL group材料进行茎秆抗性鉴定,同样发现两群体间达到了极显著差异(P0.0001),其茎杆抗性平均值分别为0.55±0.181和0.90±0.132,并最终在New-QTL group中发现了两份茎秆抗性相对于白菜亲本提高4倍多的材料,其抗性值分别为0.24±0.016和0.25±0.024,成功将C1和C3染色体上的QTLs聚合到了已含有C9染色体上的抗病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.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q943;S435.654

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