黄瓜远缘群体分子遗传连锁图谱的构建及始花节位性状的定位研究
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摘要
构建高密度的黄瓜分子遗传连锁图谱对遗传育种有着重要的研究意义,不仅是分子标记辅助选择,QTL精确定位的基础,也是基因图位克隆的关键,是加快黄瓜育种进程,提高黄瓜育种水平的重要途径。
本研究以天津科润黄瓜研究所提供的野生黄瓜品种U4和普通栽培品种Hm60-1以及由它们杂交配制得到的F2后代群体为材料,应用AFLP、SSR、SRAP等三种分子标记技术构建了黄瓜的分子遗传连锁图谱,对雌花节位性状进行了QTL定位,对黄瓜育种有着重要的指导意义。主要研究结果如下:
1.利用双亲进行引物筛选,从259对AFLP引物中筛选出有差异的引物159对,多态性比列为61.4%,其中多态性位点大于3的引物有53对;75对SRAP引物中有差异的引物为71对,多态性比列为94.6%,其中多态性好的引物有20对;113对SSR引物中,有差异的引物仅14对,多态性比列为12.4%。
2.对AFLP、SRAP、SSR等三种分子标记的多态性进行了分析,AFLP的53对引物共扩增出160个多态性位点,平均每对引物3.2个;20对SRAP引物共产生58的多态性位点,平均每对引物2.9个;在亲本间有多态性的14对SSR引物得到了14个多态性位点,平均每对引物1个,得出三者的多态性大小为:AFLP>SRAP>SSR。
3.利用JoinMap3.0对得到的160个AFLP标记,58个SRAP标记,14个SSR标记进行连锁分析,得到了一个包含10个连锁群的遗传图谱,其中包括112个AFLP标记,39个SRAP标记,8个SSR标记,该图谱覆盖基因组743.11cM,平均图距为4.67cM,每个连锁群上标记数在3～41之间,长度在8.02～140.16cM之间。
4.利用Map QTL 4.0区间作图法对雌花节位性状进行定位,共找到8个控制雌花节位的QTL位点:ff-1、ff-2、ff-3、ff-4、ff-5、ff-6、ff-7、ff-8,分别位于LG2、LG3、LG6、LG7连锁群上,这些QTL位点对雌花节位性状表型变异的贡献率分别为81.6%、81.0%、81.0%、80.0%、80.6%、81.1%、81.0%、80.7%,其中ff-1、ff-2、ff-5、ff-6、ff-7、ff-8加性效应值为正,为增效加性效应,ff-3、ff-4加性效应值为负,是减效加性效应。
A high-density molecular genetic linkage map holds an important status in cumcumber breeding. It helps molecular assisted selection, QTL mapping and map based gene cloning. In our research, a molecular genetic linakage map of cucumber were constructed by taking parents U4 ,Hm60-1 and their cross F2 population as materials. Parent U4 is a wide cultivar which found in china, Hm60-1 is a common cultivar which cultivated widely. The parents are different from each other distinctly in plant size, branches, the first female flower node, disease resistance et al.
The AFLP markers, SRAP markes, SSR markers were employed in the construction of the genetic linkage map, and Joinmap 3.0 software was used to construct the map in this study. The first female flower node trait was mapped on this linkage map by QTL mapper 4.0 software. The main results of this study were as follows:
1. 159 polymorphic primer combinations between the two parents were found from 259 AFLP primer combinations, the polymorphic ratio was 61.4%; 71 polymorphic primer combinations between the two parents were found from 75 SRAP primer combinations, the polymorphic ratio was 94.6%; 14 polymorphic primer combinations between the two parents were found from 113 SSR primer combinations, the polymorphic ratio was just 12.4%.
2. The polymorphism of the AFLP markers, SRAP markers, SSR markers were analyzed. 53 AFLP primer combinations generated 160 polymorphic locus, 3.2 per primer combination; 20 SRAP primer combinations generated 58 polymorphic locus, 2.9 per primer combination; 14 SSR pimer combinations generated 14 polymorphic locus, 1 per primer combination. The polymorphism is AFLP>SRAP>SSR.
3. Using Joinmap3.0 software, a molecular genetic linkage map which included 10 linkage groups and contained 112 AFLP markers, 39 SRAP markers, 8 SSR markers respectively was obtained. This map covering the cumcumber genome 743.11cM, the average distance between two markers is 4.67cM, which can be used for QTL mapping.
4. Mapping of the quantitative trait loci of the first female flower node was executed by Interval mapping in QTL mapper 4.0 software. The total 8 QTLs were found on the map, named them as ff1, ff2, ff3, ff4, ff5, ff6, ff7, ff8. ff1, ff2 located on linkage group2; ff3, ff4 located on linkage group3; ff5, ff6 located on linkage group6 and ff7, ff8 located on linkage group7. All of the QTLs showed positive additive effect except ff3, ff4 locus which showed negative additive effect.
本研究以天津科润黄瓜研究所提供的野生黄瓜品种U4和普通栽培品种Hm60-1以及由它们杂交配制得到的F2后代群体为材料,应用AFLP、SSR、SRAP等三种分子标记技术构建了黄瓜的分子遗传连锁图谱,对雌花节位性状进行了QTL定位,对黄瓜育种有着重要的指导意义。主要研究结果如下:
1.利用双亲进行引物筛选,从259对AFLP引物中筛选出有差异的引物159对,多态性比列为61.4%,其中多态性位点大于3的引物有53对;75对SRAP引物中有差异的引物为71对,多态性比列为94.6%,其中多态性好的引物有20对;113对SSR引物中,有差异的引物仅14对,多态性比列为12.4%。
2.对AFLP、SRAP、SSR等三种分子标记的多态性进行了分析,AFLP的53对引物共扩增出160个多态性位点,平均每对引物3.2个;20对SRAP引物共产生58的多态性位点,平均每对引物2.9个;在亲本间有多态性的14对SSR引物得到了14个多态性位点,平均每对引物1个,得出三者的多态性大小为:AFLP>SRAP>SSR。
3.利用JoinMap3.0对得到的160个AFLP标记,58个SRAP标记,14个SSR标记进行连锁分析,得到了一个包含10个连锁群的遗传图谱,其中包括112个AFLP标记,39个SRAP标记,8个SSR标记,该图谱覆盖基因组743.11cM,平均图距为4.67cM,每个连锁群上标记数在3～41之间,长度在8.02～140.16cM之间。
4.利用Map QTL 4.0区间作图法对雌花节位性状进行定位,共找到8个控制雌花节位的QTL位点:ff-1、ff-2、ff-3、ff-4、ff-5、ff-6、ff-7、ff-8,分别位于LG2、LG3、LG6、LG7连锁群上,这些QTL位点对雌花节位性状表型变异的贡献率分别为81.6%、81.0%、81.0%、80.0%、80.6%、81.1%、81.0%、80.7%,其中ff-1、ff-2、ff-5、ff-6、ff-7、ff-8加性效应值为正,为增效加性效应,ff-3、ff-4加性效应值为负,是减效加性效应。
A high-density molecular genetic linkage map holds an important status in cumcumber breeding. It helps molecular assisted selection, QTL mapping and map based gene cloning. In our research, a molecular genetic linakage map of cucumber were constructed by taking parents U4 ,Hm60-1 and their cross F2 population as materials. Parent U4 is a wide cultivar which found in china, Hm60-1 is a common cultivar which cultivated widely. The parents are different from each other distinctly in plant size, branches, the first female flower node, disease resistance et al.
The AFLP markers, SRAP markes, SSR markers were employed in the construction of the genetic linkage map, and Joinmap 3.0 software was used to construct the map in this study. The first female flower node trait was mapped on this linkage map by QTL mapper 4.0 software. The main results of this study were as follows:
1. 159 polymorphic primer combinations between the two parents were found from 259 AFLP primer combinations, the polymorphic ratio was 61.4%; 71 polymorphic primer combinations between the two parents were found from 75 SRAP primer combinations, the polymorphic ratio was 94.6%; 14 polymorphic primer combinations between the two parents were found from 113 SSR primer combinations, the polymorphic ratio was just 12.4%.
2. The polymorphism of the AFLP markers, SRAP markers, SSR markers were analyzed. 53 AFLP primer combinations generated 160 polymorphic locus, 3.2 per primer combination; 20 SRAP primer combinations generated 58 polymorphic locus, 2.9 per primer combination; 14 SSR pimer combinations generated 14 polymorphic locus, 1 per primer combination. The polymorphism is AFLP>SRAP>SSR.
3. Using Joinmap3.0 software, a molecular genetic linkage map which included 10 linkage groups and contained 112 AFLP markers, 39 SRAP markers, 8 SSR markers respectively was obtained. This map covering the cumcumber genome 743.11cM, the average distance between two markers is 4.67cM, which can be used for QTL mapping.
4. Mapping of the quantitative trait loci of the first female flower node was executed by Interval mapping in QTL mapper 4.0 software. The total 8 QTLs were found on the map, named them as ff1, ff2, ff3, ff4, ff5, ff6, ff7, ff8. ff1, ff2 located on linkage group2; ff3, ff4 located on linkage group3; ff5, ff6 located on linkage group6 and ff7, ff8 located on linkage group7. All of the QTLs showed positive additive effect except ff3, ff4 locus which showed negative additive effect.
引文
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