陆地棉优质纤维QTL的分子标记筛选及其定位
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摘要
棉花是世界上最重要的天然纤维作物,也是重要的油料作物之一。21世纪,随着纺织工业加工速度的加快和人民生活水平的提高,对棉纤维品质的要求愈来愈高。在高产、稳产的基础上,高品质棉育种成为棉花育种的主要目标之一。筛选到与优质性状中的主效基因紧密连锁的分子标记进行辅助选择,在棉花生长的早期阶段或早期分离世代就能追踪这个重要性状,可大大提高纤维品质的选择效率,加速高品质育种的进程。
本文利用01525、J415和TM-1等3个纤维品质显著差异的陆地棉材料,构建了两个F_2分离群体。通过分子标记技术和F_2分离群体构建具有较高覆盖率的陆地棉品种间的分子标记遗传图谱,进而发掘01525、J415和TM-1上与纤维品质相关的数量性状基因位点。
主要研究结果如下:
1.利用陆地棉品种01525、优质纤维材料J415和陆地棉遗传标准系TM-1作为研究材料,配置了(01525×J415)F_2和(01525×TM-1)F_2两个分离群体,使用本实验室开发或合成的SSR引物对亲本进行多态性筛选,分别获得213和128个多态性位点。在(01525×J415)F_2中,150个多态性位点构建了总长为1161.1cM的遗传图谱,该图谱由28个连锁群组成,除一个连锁群未与任何染色体对应外,其余27个与22条染色体相对应,覆盖棉花基因组的23%。在(01525×TM-1)F_2中,90个多态性位点构建了总长为1011.3cM的遗传图谱上,该图谱由24个连锁群组成,除两个连锁群未与任何染色体对应外,其余22个与17条染色体相对应,覆盖棉花基因组的22.48%。利用Joinmap 3.0完成两张图谱的整合,获得了一张包含了214个多态性位点、总长为1762.03cM的新陆地棉种内遗传连锁图谱,该图谱由39个连锁群组成,与23条染色体相对应,有两个连锁群未与任何染色体对应,覆盖棉花全基因组的35.38%,标记间平均距离8.23cM。
2.应用复合区间作图法分析了两个组合的F_2单株和F_(2:3)家系纤维品质性状,在LOD≥2.0的水平下,总共检测到66个纤维品质数量性状基因座(QTL)。包括11个纤维长度QTL,10个纤维比强度QTL,13个马克隆值QTL,12个整齐度QTL,5个短纤维率QTL及15个伸长率QTL,解释各性状表型变异的范围分别为5.3%-21.9%、4.5%-24.5%、5.8%-15.7%、5.2%-34.2%、6.1%-15.7%和5.3%-31.5%。其中9个稳定的QTL可以同时在F_2和F_(2:3)检测到,有5个QTL能在两个群体中同时检测到。两个群体增效基因的方向都与期望的表型值基本一致,大部分纤维品质的增效基因都来自于优质亲本。从QTL成簇分布和纤维品质相关分析进一步说明,纤维品质性状之间存在显著的相关性。
3.研究发现在D1、D2、A6、D6、D8、A9、A10、D10和A13上,纤维品质相关的QTL成簇分布,结合本实验室已鉴定的不同来源优质QTL的染色体定位结果,发现D6、D8、A10和A13是优质QTL的富集区。
研究结果为分析不同来源优质QTL分布特点和合理利用分子标记技术选育优质纤维品种提供了参考依据。
Cotton is the most important textile fiber crop and the world's second-most important oil-seed crop after soybean.Demands for enhancement of fiber quality traits such as fiber length and strength have been increasing because of changes in spinning technology in the textile industry.The primary breeding goal for the worldwide cotton researcher is how to genetically improve both yield and fiber quality.The development on molecular marker technology offer plant breeders a powerful tool in breeding.Especially,if the marker linked tightly with major gene controlling interesting traits is identified,the marker would allow cotton breeders to trace this very important trait in early plant-growing stage or early separation generation,and selection assisted with the marker will improve the efficiency of selection.
In this paper,molecular markers and two F_2 segregation populations derived from three parents were used to develop genetic linkage map and to exploit QTL concerned with fiber quality using upland cotton accessions 01525,J415 and TM-1,whose fiber quality is significantly different among them.
The main results are followed:
1.Two genetic linkage maps were constructed by the F_2 segregation population derived from 01525 hybridized with the J415,a high quality cotton accession and TM-1,a genetic standard line,respectively.By screening 6771 pairs of SSR primers,213 and 128 polymorphic loci were identified,respectively.Using(01525×J415)F_2 population,the genetic map is comprised of 28 linkage groups with 150 mapped loci,which could homologous with 22 chromosomes,except one linkage groups unknown to any chromosome.The 150 loci had a total genetic distance of 1161.1 cM,and covered 23%of the whole cotton genome.Using(01525×TM-1) F_2 population,the genetic map comprised of 24 linkage groups with 90 mapped loci was constructed,and could homologous with 17 chromosomes,except two unknown with any chromosome.The map had a total genetic distance of 1011.3 cM,and covered 22.48%of the whole cotton genome.A joinmap linkage map was integrated for the two F2 with SSR markers and JOINMAP3.0 software. The new integrated map is comprised of 39 linkage groups with 214 mapped loci,and could homologous with 23 chromosomes,with remained two linkage groups fail to be assigned to any chromosome.The 214 loci covered the genetic distance of 1762.03 cM, which was approximately 35.38%of the total recombination length of the cotton genome. The average distance between loci was 8.23 cM genome wide.
2.The fiber characteristics QTL of the two F_2 populations were screened using composite interval mapping(CIM) method and 66 putative quantitative trait loci(QTL) were detected for 6 fiber qualities traits,11 for fiber length,10 for fiber strength,13 for fiber micronaire,12 for fiber uniformity,5 for fiber short index,and 15 for fiber elongation. These QTL explained their corresponding phenotypic variations with 5.3%-21.9%、4.5 %-24.5%、5.8%-15.7%、5.2%-34.2%、6.1%-15.7%and 5.3%-31.5%,respectively. Among them,9 stable QTL could be found in F_2 and F_(2:3) simultaneously.At least 5 QTL could be identified at two different populations and different environments.It is also revealed that the direction of addition gene was consistent with the represent of expected value in two populations,which is inclined to the elite parents.We analyzed the relationship between clustered QTL and fiber quality,and it is also displayed to have singnificant correlation between them.
3.QTL were clustered on Chromosome D1,Chromosome D2,Chromosome A6, Chromosome D6,Chromosome D8,Chromosome A9,Chromosome A10,Chromosome D10 and Chromosome A13.Combining with the QTL tagging results fi:om different elite fiber lines in the lab,many QTL associated with fiber quality were rich in Chromosome D6, Chromosome D8,Chromosome A10 and Chromosome A13.
These results will provide references for the distribution of different QTL and utilizing the fiber quality QTL reasonably in fiber quality breeding improvement in Upland cotton.
本文利用01525、J415和TM-1等3个纤维品质显著差异的陆地棉材料,构建了两个F_2分离群体。通过分子标记技术和F_2分离群体构建具有较高覆盖率的陆地棉品种间的分子标记遗传图谱,进而发掘01525、J415和TM-1上与纤维品质相关的数量性状基因位点。
主要研究结果如下:
1.利用陆地棉品种01525、优质纤维材料J415和陆地棉遗传标准系TM-1作为研究材料,配置了(01525×J415)F_2和(01525×TM-1)F_2两个分离群体,使用本实验室开发或合成的SSR引物对亲本进行多态性筛选,分别获得213和128个多态性位点。在(01525×J415)F_2中,150个多态性位点构建了总长为1161.1cM的遗传图谱,该图谱由28个连锁群组成,除一个连锁群未与任何染色体对应外,其余27个与22条染色体相对应,覆盖棉花基因组的23%。在(01525×TM-1)F_2中,90个多态性位点构建了总长为1011.3cM的遗传图谱上,该图谱由24个连锁群组成,除两个连锁群未与任何染色体对应外,其余22个与17条染色体相对应,覆盖棉花基因组的22.48%。利用Joinmap 3.0完成两张图谱的整合,获得了一张包含了214个多态性位点、总长为1762.03cM的新陆地棉种内遗传连锁图谱,该图谱由39个连锁群组成,与23条染色体相对应,有两个连锁群未与任何染色体对应,覆盖棉花全基因组的35.38%,标记间平均距离8.23cM。
2.应用复合区间作图法分析了两个组合的F_2单株和F_(2:3)家系纤维品质性状,在LOD≥2.0的水平下,总共检测到66个纤维品质数量性状基因座(QTL)。包括11个纤维长度QTL,10个纤维比强度QTL,13个马克隆值QTL,12个整齐度QTL,5个短纤维率QTL及15个伸长率QTL,解释各性状表型变异的范围分别为5.3%-21.9%、4.5%-24.5%、5.8%-15.7%、5.2%-34.2%、6.1%-15.7%和5.3%-31.5%。其中9个稳定的QTL可以同时在F_2和F_(2:3)检测到,有5个QTL能在两个群体中同时检测到。两个群体增效基因的方向都与期望的表型值基本一致,大部分纤维品质的增效基因都来自于优质亲本。从QTL成簇分布和纤维品质相关分析进一步说明,纤维品质性状之间存在显著的相关性。
3.研究发现在D1、D2、A6、D6、D8、A9、A10、D10和A13上,纤维品质相关的QTL成簇分布,结合本实验室已鉴定的不同来源优质QTL的染色体定位结果,发现D6、D8、A10和A13是优质QTL的富集区。
研究结果为分析不同来源优质QTL分布特点和合理利用分子标记技术选育优质纤维品种提供了参考依据。
Cotton is the most important textile fiber crop and the world's second-most important oil-seed crop after soybean.Demands for enhancement of fiber quality traits such as fiber length and strength have been increasing because of changes in spinning technology in the textile industry.The primary breeding goal for the worldwide cotton researcher is how to genetically improve both yield and fiber quality.The development on molecular marker technology offer plant breeders a powerful tool in breeding.Especially,if the marker linked tightly with major gene controlling interesting traits is identified,the marker would allow cotton breeders to trace this very important trait in early plant-growing stage or early separation generation,and selection assisted with the marker will improve the efficiency of selection.
In this paper,molecular markers and two F_2 segregation populations derived from three parents were used to develop genetic linkage map and to exploit QTL concerned with fiber quality using upland cotton accessions 01525,J415 and TM-1,whose fiber quality is significantly different among them.
The main results are followed:
1.Two genetic linkage maps were constructed by the F_2 segregation population derived from 01525 hybridized with the J415,a high quality cotton accession and TM-1,a genetic standard line,respectively.By screening 6771 pairs of SSR primers,213 and 128 polymorphic loci were identified,respectively.Using(01525×J415)F_2 population,the genetic map is comprised of 28 linkage groups with 150 mapped loci,which could homologous with 22 chromosomes,except one linkage groups unknown to any chromosome.The 150 loci had a total genetic distance of 1161.1 cM,and covered 23%of the whole cotton genome.Using(01525×TM-1) F_2 population,the genetic map comprised of 24 linkage groups with 90 mapped loci was constructed,and could homologous with 17 chromosomes,except two unknown with any chromosome.The map had a total genetic distance of 1011.3 cM,and covered 22.48%of the whole cotton genome.A joinmap linkage map was integrated for the two F2 with SSR markers and JOINMAP3.0 software. The new integrated map is comprised of 39 linkage groups with 214 mapped loci,and could homologous with 23 chromosomes,with remained two linkage groups fail to be assigned to any chromosome.The 214 loci covered the genetic distance of 1762.03 cM, which was approximately 35.38%of the total recombination length of the cotton genome. The average distance between loci was 8.23 cM genome wide.
2.The fiber characteristics QTL of the two F_2 populations were screened using composite interval mapping(CIM) method and 66 putative quantitative trait loci(QTL) were detected for 6 fiber qualities traits,11 for fiber length,10 for fiber strength,13 for fiber micronaire,12 for fiber uniformity,5 for fiber short index,and 15 for fiber elongation. These QTL explained their corresponding phenotypic variations with 5.3%-21.9%、4.5 %-24.5%、5.8%-15.7%、5.2%-34.2%、6.1%-15.7%and 5.3%-31.5%,respectively. Among them,9 stable QTL could be found in F_2 and F_(2:3) simultaneously.At least 5 QTL could be identified at two different populations and different environments.It is also revealed that the direction of addition gene was consistent with the represent of expected value in two populations,which is inclined to the elite parents.We analyzed the relationship between clustered QTL and fiber quality,and it is also displayed to have singnificant correlation between them.
3.QTL were clustered on Chromosome D1,Chromosome D2,Chromosome A6, Chromosome D6,Chromosome D8,Chromosome A9,Chromosome A10,Chromosome D10 and Chromosome A13.Combining with the QTL tagging results fi:om different elite fiber lines in the lab,many QTL associated with fiber quality were rich in Chromosome D6, Chromosome D8,Chromosome A10 and Chromosome A13.
These results will provide references for the distribution of different QTL and utilizing the fiber quality QTL reasonably in fiber quality breeding improvement in Upland cotton.
引文
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