基于SSSLs的QTL鉴定及抗玉米SCMV标记开发
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摘要
本试验以我国优良的玉米杂交种豫玉22号的一个亲本自交系综3为供体亲本、另一个亲本自交系87-1为受体亲本,通过杂交、回交和SSR标记跟踪检测供体染色体片段,构建了以87-1为背景的综3的染色体单片段代换系群体。利用其中的部分纯合的单片段代换系材料对玉米的产量及产量相关性状进行了QTL鉴定。同时,基于玉米矮花叶病抗病“一致性”QTL区间查询了抗病基因的功能保守序列,根据抗感材料同源克隆序列的多态性开发了可用于玉米矮花叶病抗性鉴定的分子标记,进一步利用已知抗性的自交系和分离群体对新开发标记的有效性进行了验证,并建立了相应标记的高效、快速检测体系。主要结果如下:
1、本试验获得了57份以玉米自交系87-1为背景的综3染色体单片段代换系。57个供体代换片段不均匀地分布在玉米的10条染色体上,代换片段长度在1.75~68.35cM之间,代换片段总长度为1111.45cM,平均长度为19.50cM。覆盖基因组长度为907.70cM,覆盖率为39.09%。
2、利用获得的玉米染色体单片段代换系材料,在郑州和三亚两个试验点进行了玉米产量及产量相关性状的QTL鉴定,其中郑州点在22个单片段代换系中检出了30个QTL,三亚点在39个单片段代换系中检出了53个QTL。两个试验点检测到的玉米产量及产量相关性状的QTL总数为83个;平均每个代换片段上检出了1.73个QTL,平均每个性状检出了11.86个QTL。其中,42个QTL座位在两试验点同时检测到,占QTL座位总数的50.6%,平均每个QTL的重复检出的次数为3.23个。
3、基于玉米Bin3.04-3.05、Bin6.00-6.01区域的抗甘蔗花叶病毒“一致性”QTL区间,寻找抗病基因的功能保守域,设计特异引物,在抗感材料中扩增保守域序列,依据序列多态性开发出抗病分子标记InDel-186和InDel-130、InDel-110,对已知抗性的102份玉米自交系、BC2F2群体[(掖478ⅹ齐319)×掖478]的379个单株及重组近交系群体(X178ⅹB73)的183个家系的标记验证结果表明,InDel-186和InDel-110为共显性标记,与甘蔗花叶病毒抗性高度相关。
Many important agronomic traits are quantitative. It is very important to accurately identify and finely map quantitative trait loci (QTL). In our experiment, a series of single segment Substitution lines (SSSLs) in an elite inbred line 87-1 gnetic background of maize was developed. QTL mapping with the SSSLs at Zhengzhou and Sanya in 2008. In this study, conserved domains of disease-resistant genes were investigated within the consensus SCMV-resistance QTL on the maize bin 6.00-6.01, and furthermore the polymorphic sequences were used to develop markers InDel-130 and InDel-110. The main results are as follows:
1. A series of chromosome single segment substation lines was developed, included 57 SSSLs with one homogeneous donor segment and recipient background was developed. The 57 SSSLs were distributed on all 10 chromosomes unevenly. In the 57 SSSLs, the donor segment length is 1111.45 cM in total, the average lengenth was 19.50 cM , the maize coverd genome length was 907.70cM, the coverage was 39.09% of maize genome.
2. QTL mapping of grain yield and its components with the SSSLs at Zhengzhou and Sanya in 2008. 30 QTL was detected in 22 SSSLs in Zhengzhou, and 53 QTL was detected in 39 SSSLs in Sanya, the total QTLs was 83 in two sites. The average of each chromosome was was detected with 8.3 SSSLs, each segment was 1.73, each character was 11.86. Compare the same SSSL material in two sites, .it was found that 56 QTLs was detected in the same 17 SSSLs, and 42 QTLs was found in two sites, the percentage was 75% , the average of each repeat SSSLs was 2.47.
3. Conserved domains of disease-resistant genes were investigated within the consensus SCMV-resistance QTL on the maize bin 3.04-3.05 and bin 6.00-6.01, and furthermore the polymorphic sequences were used to develop markers InDel-186 and InDel-130, InDel-110 and susceptible marker genotypes in 102 inbred lines, It was investigated in 379 individuals of [(Ye478×Hai9-21)×Ye478] BC2F2 population and 183 lines of (X178xB73) RIL population evaluated under field condition with artificial inoculation, respectively.Furthermore, the DNA extraction, PCR amplification, and electrophoresis procedures were optimized for two markers InDel-186 and InDel-110. As a result, InDel-186、InDel-110 that was significantly associated with SCMV resistance. It was suggested that the maize breeding materials with SCMV resistance would be effectively obtained by marker-assisted selection approach with InDel-186 and InDel-110.
1、本试验获得了57份以玉米自交系87-1为背景的综3染色体单片段代换系。57个供体代换片段不均匀地分布在玉米的10条染色体上,代换片段长度在1.75~68.35cM之间,代换片段总长度为1111.45cM,平均长度为19.50cM。覆盖基因组长度为907.70cM,覆盖率为39.09%。
2、利用获得的玉米染色体单片段代换系材料,在郑州和三亚两个试验点进行了玉米产量及产量相关性状的QTL鉴定,其中郑州点在22个单片段代换系中检出了30个QTL,三亚点在39个单片段代换系中检出了53个QTL。两个试验点检测到的玉米产量及产量相关性状的QTL总数为83个;平均每个代换片段上检出了1.73个QTL,平均每个性状检出了11.86个QTL。其中,42个QTL座位在两试验点同时检测到,占QTL座位总数的50.6%,平均每个QTL的重复检出的次数为3.23个。
3、基于玉米Bin3.04-3.05、Bin6.00-6.01区域的抗甘蔗花叶病毒“一致性”QTL区间,寻找抗病基因的功能保守域,设计特异引物,在抗感材料中扩增保守域序列,依据序列多态性开发出抗病分子标记InDel-186和InDel-130、InDel-110,对已知抗性的102份玉米自交系、BC2F2群体[(掖478ⅹ齐319)×掖478]的379个单株及重组近交系群体(X178ⅹB73)的183个家系的标记验证结果表明,InDel-186和InDel-110为共显性标记,与甘蔗花叶病毒抗性高度相关。
Many important agronomic traits are quantitative. It is very important to accurately identify and finely map quantitative trait loci (QTL). In our experiment, a series of single segment Substitution lines (SSSLs) in an elite inbred line 87-1 gnetic background of maize was developed. QTL mapping with the SSSLs at Zhengzhou and Sanya in 2008. In this study, conserved domains of disease-resistant genes were investigated within the consensus SCMV-resistance QTL on the maize bin 6.00-6.01, and furthermore the polymorphic sequences were used to develop markers InDel-130 and InDel-110. The main results are as follows:
1. A series of chromosome single segment substation lines was developed, included 57 SSSLs with one homogeneous donor segment and recipient background was developed. The 57 SSSLs were distributed on all 10 chromosomes unevenly. In the 57 SSSLs, the donor segment length is 1111.45 cM in total, the average lengenth was 19.50 cM , the maize coverd genome length was 907.70cM, the coverage was 39.09% of maize genome.
2. QTL mapping of grain yield and its components with the SSSLs at Zhengzhou and Sanya in 2008. 30 QTL was detected in 22 SSSLs in Zhengzhou, and 53 QTL was detected in 39 SSSLs in Sanya, the total QTLs was 83 in two sites. The average of each chromosome was was detected with 8.3 SSSLs, each segment was 1.73, each character was 11.86. Compare the same SSSL material in two sites, .it was found that 56 QTLs was detected in the same 17 SSSLs, and 42 QTLs was found in two sites, the percentage was 75% , the average of each repeat SSSLs was 2.47.
3. Conserved domains of disease-resistant genes were investigated within the consensus SCMV-resistance QTL on the maize bin 3.04-3.05 and bin 6.00-6.01, and furthermore the polymorphic sequences were used to develop markers InDel-186 and InDel-130, InDel-110 and susceptible marker genotypes in 102 inbred lines, It was investigated in 379 individuals of [(Ye478×Hai9-21)×Ye478] BC2F2 population and 183 lines of (X178xB73) RIL population evaluated under field condition with artificial inoculation, respectively.Furthermore, the DNA extraction, PCR amplification, and electrophoresis procedures were optimized for two markers InDel-186 and InDel-110. As a result, InDel-186、InDel-110 that was significantly associated with SCMV resistance. It was suggested that the maize breeding materials with SCMV resistance would be effectively obtained by marker-assisted selection approach with InDel-186 and InDel-110.
引文
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