玉米抗灰斑病主效QTL发掘及其精细定位
发布时间：2018-09-10 20:40
【摘要】：由尾孢菌侵染玉米叶片引起的玉米灰斑病已成为我国东华北及西南玉米产区重要的病害之一,发掘抗灰斑病基因并应用于抗病品种选育是经济有效的防控途径。本文首先通过收集并整理玉米抗灰斑病QTL文献,利用元分析方法进行整合,确定"一致性"抗病QTL;然后以CN165为供体亲本和81162为受体亲本构建的片段代换系群体,进行QTL验证,发掘主效QTL位点;再筛选含有目标区段的片段代换系构建分离群体,进行QTL精细定位。取得的主要研究结果如下:利用元分析方法分析并整理了 14篇玉米抗灰斑病QTL文献的信息,共筛选确定了13个一致性QTL区间,分布在玉米第1、2、4、7、8号染色体上。利用以自交系81162为轮回亲本、自交系CN165为非轮回亲本构建的回交导入群体根据连锁不平衡原理对13个一致性QTL进行验证,在13个一致性QTL区间共获得20个偏分离位点。第1和第4染色体上偏分离最严重,其他染色体上偏分离度较小。说明第1和第4染色体上存在着效应较大的抗病QTL。第1染色体标记umc2227、bnlg1832、umc1243、umc2025、umc1515、umc1297、umc1461 处供体基因频率均在 50%以上,可能存在几个连锁的抗病基因。第4染色体上基因位于标记bnlg2291和umc1 194之间。结合背景检测结果,从BC2F3抗病家系中选取3个目标家系,与轮回亲本81162杂交并自交得到目标区段分离群体。选取来自同一自交果穗的176个单株,采用完备区间作图法,对第4染色体的主效QTL-CNgls4进行定位,将QTL定位于标记umc1329-bnlg1137 之间。根据B73序列信息自主开发引物67对,在目标区段获得多态性引物7对,利用这7对多态性引物对分离群体的882个高抗和37个高感单株进行重组体筛选,在该区间内共找到11个重组体。通过分析,将QTL区间精细定位到标记Snyw042和Snyw008之间,物理距离为2.19Mb,物理位置为 167,355,376bp-169,548,333bp。通过发掘自交系CN165中第4染色体的主效QTL,为选育抗玉米灰斑病的种质材料、玉米分子标记辅助选择育种及抗灰斑病基因的克隆和功能分析提供理论参考。
[Abstract]:The gray spot of maize caused by the infection of cemocystis in maize leaves has become one of the most important diseases in maize growing areas in the east and southwest of China. It is an economical and effective way to explore the genes of resistance to gray spot disease and to apply it to the breeding of disease-resistant varieties. In this paper, the QTL literature on resistance to gray spot disease in maize was collected and sorted, and the "consistent" disease-resistant QTL; was determined by meta-analysis method. Then the fragment substitution line population constructed with CN165 as donor parent and 81162 as recipient parent was used for QTL verification. The dominant QTL loci were identified, and then the fragment substitution lines containing the target region were selected to construct isolated populations for QTL precise mapping. The main results were as follows: 14 papers on QTL resistance to gray spot disease in maize were analyzed and sorted by meta-analysis. Thirteen consistent QTL intervals were selected and distributed on chromosome 7 and 8 in maize. Using inbred line 81162 as recurrent parent and inbred line CN165 as non-recurrent parent, 13 consistent QTL were verified according to linkage disequilibrium principle. A total of 20 partial segregation sites were obtained in 13 consistent QTL intervals. The first and fourth chromosomes were most seriously biased, while the other chromosomes were less skewed. It is suggested that there are disease-resistant QTL. on chromosomes 1 and 4. The frequency of donor genes at umc2227,bnlg1832,umc1243,umc2025,umc1515,umc1297,umc1461 on chromosome 1 was above 50%, and there might be several linkage resistant genes. The genes on chromosome 4 are located between the labeled bnlg2291 and umc1 194. Based on the results of background test, three target families were selected from BC2F3 resistant families, crossed with recurrent parent 81162 and self-crossed to obtain the target segment segregation population. 176 individual plants from the same self-pollinated ear were selected and the main effect QTL-CNgls4 of chromosome 4 was mapped by complete interval mapping method. The QTL was mapped between labeled umc1329-bnlg1137. According to the information of B73 sequence, 67 pairs of primers were developed and 7 pairs of polymorphic primers were obtained in the target region. The 7 pairs of polymorphic primers were used to screen 882 highly resistant and 37 highly susceptible individual plants from isolated populations. A total of 11 recombinant bodies were found in this region. Through the analysis, the QTL interval was finely located between the tagged Snyw042 and Snyw008, the physical distance was 2.19 Mb, and the physical position was 167355376bp-169548333bp. The main effect QTL, of chromosome 4 in inbred line CN165 was found to be a germplasm material for breeding resistant to gray spot of maize. Molecular marker-assisted selection breeding of maize and cloning and functional analysis of genes resistant to gray spot disease were provided as theoretical references.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S435.13
，
本文编号：2235553
[Abstract]:The gray spot of maize caused by the infection of cemocystis in maize leaves has become one of the most important diseases in maize growing areas in the east and southwest of China. It is an economical and effective way to explore the genes of resistance to gray spot disease and to apply it to the breeding of disease-resistant varieties. In this paper, the QTL literature on resistance to gray spot disease in maize was collected and sorted, and the "consistent" disease-resistant QTL; was determined by meta-analysis method. Then the fragment substitution line population constructed with CN165 as donor parent and 81162 as recipient parent was used for QTL verification. The dominant QTL loci were identified, and then the fragment substitution lines containing the target region were selected to construct isolated populations for QTL precise mapping. The main results were as follows: 14 papers on QTL resistance to gray spot disease in maize were analyzed and sorted by meta-analysis. Thirteen consistent QTL intervals were selected and distributed on chromosome 7 and 8 in maize. Using inbred line 81162 as recurrent parent and inbred line CN165 as non-recurrent parent, 13 consistent QTL were verified according to linkage disequilibrium principle. A total of 20 partial segregation sites were obtained in 13 consistent QTL intervals. The first and fourth chromosomes were most seriously biased, while the other chromosomes were less skewed. It is suggested that there are disease-resistant QTL. on chromosomes 1 and 4. The frequency of donor genes at umc2227,bnlg1832,umc1243,umc2025,umc1515,umc1297,umc1461 on chromosome 1 was above 50%, and there might be several linkage resistant genes. The genes on chromosome 4 are located between the labeled bnlg2291 and umc1 194. Based on the results of background test, three target families were selected from BC2F3 resistant families, crossed with recurrent parent 81162 and self-crossed to obtain the target segment segregation population. 176 individual plants from the same self-pollinated ear were selected and the main effect QTL-CNgls4 of chromosome 4 was mapped by complete interval mapping method. The QTL was mapped between labeled umc1329-bnlg1137. According to the information of B73 sequence, 67 pairs of primers were developed and 7 pairs of polymorphic primers were obtained in the target region. The 7 pairs of polymorphic primers were used to screen 882 highly resistant and 37 highly susceptible individual plants from isolated populations. A total of 11 recombinant bodies were found in this region. Through the analysis, the QTL interval was finely located between the tagged Snyw042 and Snyw008, the physical distance was 2.19 Mb, and the physical position was 167355376bp-169548333bp. The main effect QTL, of chromosome 4 in inbred line CN165 was found to be a germplasm material for breeding resistant to gray spot of maize. Molecular marker-assisted selection breeding of maize and cloning and functional analysis of genes resistant to gray spot disease were provided as theoretical references.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S435.13
，
本文编号：2235553
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