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

doi:10.11733/j.issn.1007-0435.2006.01.003

›› 2006, Vol. 14 ›› Issue (1): 9-13,23.DOI: 10.11733/j.issn.1007-0435.2006.01.003

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

丁成龙^1,2, 沈益新¹, 顾共如², 三浦优一³

1. 南京农业大学动物科技学院, 南京, 210095;
2. 江苏省农业科学院畜牧研究所, 南京, 210014;
3. 日本畜产种子协会饲料作物研究所, 枥木, 329-2742

收稿日期:2005-03-04 修回日期:2005-06-29 出版日期:2006-02-15 发布日期:2006-02-15
通讯作者: 沈益新,E-mail:cts-China@Sohu.com
作者简介:丁成龙(1969- ),男,副研究员,现为南京农业大学在读博士生,从事牧草育种及栽培利用研究工作,发表研究论文20余篇,E-mail:dingcl@jaas.ac.cn;
基金资助:
中日合作项目“饲料作物基因组解析”;日本竞马协会提供资助

Identification of an EST-CAPS Marker Linking to the Resistance Gene of Italian Ryegrass Gray-Leaf-Spot Disease by Bulked Segregant Analysis

DING Cheng-long^1,2, SHEN Yi-xin¹, GU Hong-ru², Yuichi Miura³

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 Province 210014, China;
3. Institute of Forage Crop, Japan Grassland Farming and Forage Seed Association, Tochigi 329-2742, Japan

Received:2005-03-04 Revised:2005-06-29 Online:2006-02-15 Published:2006-02-15

摘要/Abstract

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)

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.

Key words: Italian ryegrass, Gray leaf spot, Molecular marker, Expressed sequence tag(EST), Cleaved amplified polymorphic sequence(CAPS)

中图分类号:

S812
Q943

丁成龙, 沈益新, 顾共如, 三浦优一. 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]. , 2006, 14(1): 9-13,23.

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]. , 2006, 14(1): 9-13,23.

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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

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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