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| 分群分析法获得与多花黑麦草抗叶班病基因连锁的EST-CAPS标记 | |
| 引用本文： | 丁成龙,沈益新,顾共如,三浦 優一.分群分析法获得与多花黑麦草抗叶班病基因连锁的EST-CAPS标记[J].草地学报,2006,14(1):9-13,23. |
| 作者姓名： | 丁成龙 沈益新 顾共如 三浦 優一 |
| 作者单位： | 1. 南京农业大学动物科技学院,南京,210095;江苏省农业科学院畜牧研究所,南京,210014 2. 南京农业大学动物科技学院,南京,210095 3. 江苏省农业科学院畜牧研究所,南京,210014 4. 日本畜产种子协会饲料作物研究所,枥木,329-2742 |
| 基金项目： | 中-日合作项目 , 日本竞马协会资助项目 |
| 摘 要： | 利用多花黑麦草叶斑病抗性和敏感个体杂交构建F1分离群体,采用分群分析法(bulked segregant analysis,BSA),通过多花黑麦草表达序列标签(expressed sequence tag,EST)的PCR扩增和扩增产物的限制性内切酶酶切多态性(cleaved amplified polymorphic sequence,CAPS)筛选,获得一个同多花黑麦草草抗叶斑病紧密连锁的EST-CAPS标记p56.对照多花黑麦草细胞质雄性不育(CMS)群体构建的遗传连锁图,该EST-CAPS标记位于多花黑麦草的第5遗传连锁群(LG5).对cDNA文库中对应于标记位点p56的EST序列片段分析和基因库搜索结果表明,该EST所编码的氨基酸序列同大麦天冬酰胺合成酶基因HvAS1和HvAS2的部分氨基酸序列片段同源性很高,推测位点p56所处的基因为编码多花黑麦草天冬酰胺合成酶基因.该分子标记可用于多花黑麦草分子标记辅助育种. |
| 关 键 词： | 多花黑麦草 叶班病 分子标记 表达序列标签(EST) 酶切扩增片段多态性序列(CAPS) |
| 文章编号： | 1007-0435（2006）01-0009-06 |
| 收稿时间： | 2005-03-04 |
| 修稿时间： | 2005-03-042005-06-29 |
Identification of an EST-CAPS Marker Linking to the Resistance Gene of Italian Ryegrass Gray-Leaf-Spot Disease by Bulked Segregant Analysis |
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| DING Cheng-long,SHEN Yi-xin,GU Hong-ru,Yuichi Miura.Identification of an EST-CAPS Marker Linking to the Resistance Gene of Italian Ryegrass Gray-Leaf-Spot Disease by Bulked Segregant Analysis[J].Acta Agrestia Sinica,2006,14(1):9-13,23. | |
| Authors: | DING Cheng-long SHEN Yi-xin GU Hong-ru Yuichi Miura |
| Institution: | 1. College of Animal Science and Technology, Nanjing Agriculture University, Nanjing, Jiangsu Province 210095, China; 2. Institute of Animal Science, Jiangsu Academy of Agricultural Science, Nanjing, Jiangsu Provinee 210014, China; 3. Institute of Forage Crop, Japan Grassland Farming and Forage Seed Association, Tochigi 329 2742, Japan |
| Abstract: | An F1 separation population was formed by crossing the gray-leaf-spot resistant individual of Italian ryegrass with the susceptible one.Expressed sequence tag(EST) of Italian ryegrass were amplified from the gDNA of two parents and F1 population and digested with endonucleases to detect the polymorphisms by the method of bulked segregant analysis(BSA).An expressed sequence tag(EST)cleaved amplified polymorphic sequence(CAPS) marker,p56,which linked to gray leaf disease resistance gene of Italian ryegrass was identified.Compared to the reference genetic linkage maps of Italian ryegrass constructed from cytoplasmic male sterility,the marker p56 was mapped on the fifth linkage group(LG5).The deduced amino acid sequence of p56 EST from cDNA library was highly identical to that of gene HvAS1 and HvAS2 which coded asparagine synthetase from barley.The results revealed that the EST putatively coded asparagine synthetase from Italian ryegrass.The molecular marker could be used for marker assistance selection in Italian ryegrass breeding. |
| Keywords: | Italian ryegrass Gray leaf spot Molecular marker Expressed sequence tag(EST) Cleaved amplified polymorphic sequence(CAPS) |
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