小麦EST-SSR标记的开发和遗传作图  

Development and Mapping of EST-SSR Markers in Wheat

作　　者：潘海涛[1] 汪俊君[1] 王盈盈[1] 齐照良[1] 李斯深[1] 

机构地区：[1]山东农业大学农学院/作物生物学国家重点实验室,山东泰安271018

出　　处：《中国农业科学》2010年第3期452-461,共10页Scientia Agricultura Sinica

基　　金：国家自然科学基金(3057155)

摘　　要：【目的】利用小麦EST序列数据库开发EST-SSR标记。【方法】对GenBank/dbEST注册的普通小麦EST序列(2006.4.18—2007.2.4)进行SSR查找,采用Primer5.0软件设计EST-SSR引物,选用3个小麦品种进行有效性检测,利用RIL群体和Mapmaker/Exp3.0软件进行遗传作图。【结果】在265362条普通小麦EST序列中,共发现6314个SSR,占整个EST数据库的2.38%。其中二核苷酸、三核苷酸重复序列最多,分别为2237(35.43%)和2084(33.01%)个。二核苷酸重复中,以GA/CT和AG/TC出现频率最高、分别占SSR总数的17.85%和10.37%,其次是CA/GT(4.07%)和AC/TG(2.53%);三核苷酸重复中,CAA/GTT(3.93%)、CGG/GCC(3.83%)、CGC/GCG(3.36%)、GGC/CCG(3.14%)、CTT/GAA(2.53%)、TGC/ACG(2.27%)以较高的频率出现。根据筛选得到的微卫星序列共设计了596个EST-SSR引物对,选择其中95分以上的194个合成。PCR检测表明,165个引物对(85%)可以扩增出稳定清晰的带型;在RIL群体中检测到21个EST-SSR引物26个位点有多态性,将其中的23个位点整合到已有的小麦遗传图谱上。【结论】开发了165个小麦EST-SSR新标记,EST序列是小麦SSR标记的重要来源。[Objective] The objective of this study was to develop new simple sequence repeat （SSR） markers from EST （Expressed sequence tag） sequences （EST-SSR markers） for wheat. [Method] The wheat EST sequences from GenBank/dbEST （from April 18, 2006 to February 4, 2007） were used to search for SSRs, EST-SSR primer pairs were designed using Primer5.0 software, the validity of primer pairs were detected using three wheat varieties, and the marker loci were mapped using RIL population and Mapmaker/Exp3.0 software. [Result] From the 265 362 wheat EST sequences released in GenBank/dbEST, 6 314 SSRs were found which amounts to 2.38% of the total number of ESTs. Among those EST-SSRs, the most motifs were dinucleotides and trinucleotides, which were 2 237 （35.43%） and 2 084 （33.01%）, respectively. For dinucleotides, GA/CT （17.85%） and AG/TC （10.37%） were the most frequent repeat, followed by CA/GT （4.07%） and AC/TG （2.53%）; and for trinucleotides, CAA/GTT （3.93%）, CGG/GCC （3.83%）, CGC/GCG （3.36%）, GGC/CCG （3.14%）, CTT/GAA （2.53%） and TGC/ACG （2.27%） appeared high frequency. Based on these EST-SSR sequences, 596 EST-SSR primer pairs were designed, 194 of which were synthesized, and 165 （85%） led to PCR amplification products. Using a RIL population, the polymorphism of 21 EST-SSR primer pairs with 26 loci were detected, and 23 of those loci were integrated on the wheat genetic map. [Conclusion] A total of 165 new wheat EST-SSR markers have been developed. EST sequences are important sources for developing SSR markers.

关 键 词：小麦 EST—SSR 分子标记 RIL 遗传图谱 

分 类 号：S512.1[农业科学—作物学]