黄颡鱼（Pelteobagrus fulvidraco）遗传图谱构建及生长相关性状的QTL定位
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摘要
本研究采用拟测交策略,以野生(♂)和人工养殖(♀)黄颡鱼人工授精产生的100个F1个体为作图群体,用SSR、SRAP和TRAP三种DNA分子标记技术构建了黄颡鱼的遗传连锁图谱,并对5个与生长相关的性状进行了QTL分析。
从50对SSR引物中扩增得到13个在双亲间呈现多态性并在子代分离的标记,其中母本标记4个,父本标记3个,母本和父本共有标记6个。卡方检验显示,13个微卫星标记在子代中的分离全部符合其期望比例。
从196对SRAP引物中扩增得到89个标记,平均每对引物产生1.3个标记,其中母本标记38个,父本标记46个,母本和父本共有标记5个。卡方检验显示,母本标记中有8个发生偏分离,父本标记中有11个发生偏分离。子代分离符合期望比的标记占构图SRAP标记的比例为78.6%。
从56对TRAP引物中扩增得到26个标记,平均每对引物产生1.2个标记,其中母本标记9个,父本标记8个,母本和父本共有标记9个。卡方检验显示,母本标记中有2个发生偏分离,父本标记中有3个发生偏分离。子代分离符合期望比的标记占构图TRAP标记的比例为80.8%。
利用这些标记构建了黄颡鱼遗传连锁图谱。其中雌性图谱整合了51个标记,图谱的长度为585.5 cM；包括16个连锁群,连锁群的长度从8.2-127.4 cM,每个连锁的标记个数为2-8个；雌性图谱连锁群6的平均间隔最大为32.1 cM,连锁群14的平均间隔最小为4.6 cM,图谱整体平均间隔为16.3 cM。雄性图谱共有56个标记,图谱的长度为752.3 cM；包括15个连锁群,连锁群的长度从3.0-205.2 cM,每个连锁的标记个数为2-11个；雄性图谱连锁群6的平均间隔最大为28.8cM,连锁群14的平均间隔最小为5.0cM,图谱整体平均间隔为17.7 cM。共有图谱有20个标记,图谱的长度为231.3 cM；包括5个连锁群,连锁群的长度从5.3-84.7cM,每个连锁的标记个数为2-6个；共有图谱连锁群5的平均间隔最大为20.3cM,连锁群1的平均间隔最小为5.3cM,共有图谱整体平均间隔为13.8cM。估算的雌雄图谱的覆盖率分别为52.6%和58.7%。
用黄颡鱼雌雄连锁图谱对5个与其生长相关的性状进行了QTL扫描,在雌性图谱上检测到1个头宽的QTL,定位于第7连锁群上,LOD值为3.2,可解释的表性变异为13%。在雄性图谱上分别检测到1个体高和体长的QTL,均定位于第1连锁群上。体高QTL的LOD值为2.4,可解释的表性变异为12%。全长QTL的LOD值为2.1,可解释的表性变异为11%。本研究获得的3个QTL均可尝试用于黄颡鱼的生长性状标记辅助育种。
黄颡鱼遗传图谱的构建及生长相关性状的QTL定位为下一步进行黄颡鱼的分子标记辅助选择育种打下基础,并最终推动黄颡鱼的遗传改良。
A segregating population including 100 progenies from the cross between wild ((?)) and breeding (♀) yellow catfish was obtained based on a pseudo-testcross mapping strategy. Genetic linkage maps of yellow catfish (Pelteobagrus fulvidraco) were constructed by using SSR、SRAP and TRAP markers. In addition, QTLs related to five growth traits were analyzed.
Polymorphic markers segregated in one of the parents were amplified by using thirteen pairs of primers detected within totally fifty pairs of SSR primers, of which four markers derived from female parent, three markers derived from male parent and six markers from combined parents. All SSR markers segregated according to the expected Mendelian ratio.
Eighty-nine markers were amplified by using 196 pairs of SRAP primers. The average amplify amount of marker was 1.3 by each pair of primer. Thirty-eight markers were detected in female parent and forty-six markers in male parent, meanwhile five markers from combined parents. Chi-square Test shows that eight and eleven markers appear segregation distortion in female and male parent respectively. Totally markers segregated according to the expected Mendelian ratio are 78.6% in all SRAP markers.
Twenty-six markers were amplified by using 56 pairs of TRAP primers. The average amplify amount of marker was 1.2 by each pair of primer. Nine markers were detected in female parent and eight markers in male parent, meanwhile nine markers from combined parents. Chi-square Test shows that two and three markers appear segregation distortion in female and male parent respectively. Totally markers segregated according to the expected Mendelian ratio are 80.8% in all TRAP markers.
These markers were used to construct genetic map of yellow catfish. Fifty-one markers were involved in female map. It was covering a total of 585.5 cM in length and included sixteen linkage groups, the length skip from 8.2 to 127.4 cM, the average amount of marker in single linkage group skip from 2 to 8. The longest average gap distance was 32.1 cM located in the sixth linkage group and the shortest average gap distance was 4.6 cM located in the 14th linkage group. The average gap distance was 16.3 cM in the whole female map. Fifty-six markers were involved in male map. It was covering a total of 752.3 cM in length and included fifteen linkage groups, the length skip from 3.0 to 205.2 cM, the average amount of marker in single linkage group skip from 1 to 11. The longest average gap distance was 28.8 cM located in the sixth linkage group and the shortest average gap distance is 5 cM located in the 14th linkage group. The average gap distance was 17.7 cM in the whole male map. Twenty markers were involved in combined map. It was covering a total of 231.3 cM in length and included five linkage groups, the length skip from 5.3 to 84.7cM, the average amount of marker in one linkage group skip from 2 to 6. The longest average gap distance was 20.3cM located in the fifth linkage group and the shortest average gap distance was 5.3 cM located in the first linkage group. The average gap distance was 13.8 cM in the whole combined map. The coverage scale was 52.6% and 58.7% on female and male map respectively.
These maps were used to scan QTLs related to five growth traits of yellow catfish. One Head length QTL was identified in female map that located in LG7, LOD value is 3.2 and explains 13% of phenotypic variation. A Body width and a Body length QTL were identified in male map that all located in LG1, LOD value is 2.4 and 2.1 respectively, while explains 12% and 11% of phenotypic variation respectively. These QTL could be considered to apply in marker-assisted selection for growth-related traits breeding of yellow catfish.
The construction of genetic linkage maps of yellow catfish and QTL sites related to growth traits will lay foundations for Marker-assisted selection breeding of yellow catfish. It will alter the genetic improvement of yellow catfish.
从50对SSR引物中扩增得到13个在双亲间呈现多态性并在子代分离的标记,其中母本标记4个,父本标记3个,母本和父本共有标记6个。卡方检验显示,13个微卫星标记在子代中的分离全部符合其期望比例。
从196对SRAP引物中扩增得到89个标记,平均每对引物产生1.3个标记,其中母本标记38个,父本标记46个,母本和父本共有标记5个。卡方检验显示,母本标记中有8个发生偏分离,父本标记中有11个发生偏分离。子代分离符合期望比的标记占构图SRAP标记的比例为78.6%。
从56对TRAP引物中扩增得到26个标记,平均每对引物产生1.2个标记,其中母本标记9个,父本标记8个,母本和父本共有标记9个。卡方检验显示,母本标记中有2个发生偏分离,父本标记中有3个发生偏分离。子代分离符合期望比的标记占构图TRAP标记的比例为80.8%。
利用这些标记构建了黄颡鱼遗传连锁图谱。其中雌性图谱整合了51个标记,图谱的长度为585.5 cM；包括16个连锁群,连锁群的长度从8.2-127.4 cM,每个连锁的标记个数为2-8个；雌性图谱连锁群6的平均间隔最大为32.1 cM,连锁群14的平均间隔最小为4.6 cM,图谱整体平均间隔为16.3 cM。雄性图谱共有56个标记,图谱的长度为752.3 cM；包括15个连锁群,连锁群的长度从3.0-205.2 cM,每个连锁的标记个数为2-11个；雄性图谱连锁群6的平均间隔最大为28.8cM,连锁群14的平均间隔最小为5.0cM,图谱整体平均间隔为17.7 cM。共有图谱有20个标记,图谱的长度为231.3 cM；包括5个连锁群,连锁群的长度从5.3-84.7cM,每个连锁的标记个数为2-6个；共有图谱连锁群5的平均间隔最大为20.3cM,连锁群1的平均间隔最小为5.3cM,共有图谱整体平均间隔为13.8cM。估算的雌雄图谱的覆盖率分别为52.6%和58.7%。
用黄颡鱼雌雄连锁图谱对5个与其生长相关的性状进行了QTL扫描,在雌性图谱上检测到1个头宽的QTL,定位于第7连锁群上,LOD值为3.2,可解释的表性变异为13%。在雄性图谱上分别检测到1个体高和体长的QTL,均定位于第1连锁群上。体高QTL的LOD值为2.4,可解释的表性变异为12%。全长QTL的LOD值为2.1,可解释的表性变异为11%。本研究获得的3个QTL均可尝试用于黄颡鱼的生长性状标记辅助育种。
黄颡鱼遗传图谱的构建及生长相关性状的QTL定位为下一步进行黄颡鱼的分子标记辅助选择育种打下基础,并最终推动黄颡鱼的遗传改良。
A segregating population including 100 progenies from the cross between wild ((?)) and breeding (♀) yellow catfish was obtained based on a pseudo-testcross mapping strategy. Genetic linkage maps of yellow catfish (Pelteobagrus fulvidraco) were constructed by using SSR、SRAP and TRAP markers. In addition, QTLs related to five growth traits were analyzed.
Polymorphic markers segregated in one of the parents were amplified by using thirteen pairs of primers detected within totally fifty pairs of SSR primers, of which four markers derived from female parent, three markers derived from male parent and six markers from combined parents. All SSR markers segregated according to the expected Mendelian ratio.
Eighty-nine markers were amplified by using 196 pairs of SRAP primers. The average amplify amount of marker was 1.3 by each pair of primer. Thirty-eight markers were detected in female parent and forty-six markers in male parent, meanwhile five markers from combined parents. Chi-square Test shows that eight and eleven markers appear segregation distortion in female and male parent respectively. Totally markers segregated according to the expected Mendelian ratio are 78.6% in all SRAP markers.
Twenty-six markers were amplified by using 56 pairs of TRAP primers. The average amplify amount of marker was 1.2 by each pair of primer. Nine markers were detected in female parent and eight markers in male parent, meanwhile nine markers from combined parents. Chi-square Test shows that two and three markers appear segregation distortion in female and male parent respectively. Totally markers segregated according to the expected Mendelian ratio are 80.8% in all TRAP markers.
These markers were used to construct genetic map of yellow catfish. Fifty-one markers were involved in female map. It was covering a total of 585.5 cM in length and included sixteen linkage groups, the length skip from 8.2 to 127.4 cM, the average amount of marker in single linkage group skip from 2 to 8. The longest average gap distance was 32.1 cM located in the sixth linkage group and the shortest average gap distance was 4.6 cM located in the 14th linkage group. The average gap distance was 16.3 cM in the whole female map. Fifty-six markers were involved in male map. It was covering a total of 752.3 cM in length and included fifteen linkage groups, the length skip from 3.0 to 205.2 cM, the average amount of marker in single linkage group skip from 1 to 11. The longest average gap distance was 28.8 cM located in the sixth linkage group and the shortest average gap distance is 5 cM located in the 14th linkage group. The average gap distance was 17.7 cM in the whole male map. Twenty markers were involved in combined map. It was covering a total of 231.3 cM in length and included five linkage groups, the length skip from 5.3 to 84.7cM, the average amount of marker in one linkage group skip from 2 to 6. The longest average gap distance was 20.3cM located in the fifth linkage group and the shortest average gap distance was 5.3 cM located in the first linkage group. The average gap distance was 13.8 cM in the whole combined map. The coverage scale was 52.6% and 58.7% on female and male map respectively.
These maps were used to scan QTLs related to five growth traits of yellow catfish. One Head length QTL was identified in female map that located in LG7, LOD value is 3.2 and explains 13% of phenotypic variation. A Body width and a Body length QTL were identified in male map that all located in LG1, LOD value is 2.4 and 2.1 respectively, while explains 12% and 11% of phenotypic variation respectively. These QTL could be considered to apply in marker-assisted selection for growth-related traits breeding of yellow catfish.
The construction of genetic linkage maps of yellow catfish and QTL sites related to growth traits will lay foundations for Marker-assisted selection breeding of yellow catfish. It will alter the genetic improvement of yellow catfish.
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