小麦EST-SSR标记的开发和染色体定位及其在追踪黑麦染色体中的应用  

Development and Chromosome Mapping of 81 New Wheat EST-SSR Markers and Application for Characterizing Rye Chromosomes Added in Wheat

作　　者：庄丽芳[1] 宋立晓[1] 冯祎高[1] 钱保俐[1] 徐海滨[1] 裴自友[1] 亓增军[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室,江苏南京210095

出　　处：《作物学报》2008年第6期926-933,共8页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(30500048,30770222);高等学校创新引智计划项目(B08025);山西省留学基金项目(2005077)

摘　　要：根据小麦盐胁迫诱导和茎秆组织相关EST序列开发了81对EST-SSR引物,其中67、46、18和61对分别在小麦、黑麦、簇毛麦和大麦基因组中稳定扩增,在不同小麦和大麦品种间具有多态性的引物分别有22和23对。利用小麦缺体-四体系共定位了43对引物的81个位点,其中A、B和D染色体组上分别有29、30和22个位点,涉及除4B、3D和6D外的18条染色体。此外30对引物在黑麦基因组中具有特异扩增,其中8对分别在黑麦1R、4R、5R和R7染色体上具有特异扩增,7对在多条黑麦染色体具有相同扩增。这些新标记可有效用于小麦及其近缘物种的遗传作图与比较遗传研究。Genomic microsatellites （gSSR） have been widely used in cultivar fingerprinting, genetic diversity assessment, molecular mapping and marker assisted breeding, but development of SSR markers from genomic libraries is expensive and inefficient. With the availability of large numbers of expressed sequence tags （ESTs）, development of EST-derived SSR （eSSR） markers is becoming an efficient and low-cost option for many plant species. To date, numerous eSSR markers have been developed and used in comparative and genetic mapping in many plant species, which showed a higher rate of transferability than gSSR markers. In wheat （Triticum aestivum L.）, until January, 2007, more than 854 015 ESTs have been deposited in the public GenBank （http：//www.ncbi.nlm.nih.gov）, and more than 16 000 ESTs have been physically positioned on different bins of wheat chromosomes via Southern blotting analysis on a set of wheat deletions. Despite series of PCR based eSSR primers were designed, only a small number of them have been tested in wheat or its relative genomes. The purposes of our research were： 1） to develop and map of new eSSR markers of wheat; 2） to test the transferability of these markers in wheat relatives; and 3） to screen of new markers tagging different rye （Secale cereale L.） chromosomes. In this paper, 81 new EST-derived SSR markers were developed from salinity stressed and stem relative cDNA libraries of wheat. Of them, 67, 46, 18, and 61 markers produced 124, 72, 26, and 124 stable amplicons in genomes of wheat, rye, Haynaldia villosa L. Schur., and barley （Hordeum vulgare L.）, respectively, indicaring high transferability of these markers in wheat relatives. Twenty-two markers produced polymorphic amplicons between wheat landrace Chinese Spring and Huixianhong, and 23 markers produced polymorphic loci between barley culrivar Sileng Damai and Kuaiying Luomai, indicating potential use of these markers in genetic mapping and fingerprinting. Among the 67 eSSR markers with stable amplico

关 键 词：小麦 EST-SSR 染色体定位 黑麦 专化性标记 

分 类 号：S512.1[农业科学—作物学] S513