适用于大豆疫霉菌遗传分析的新EST-SSR标记  

Novel EST-SSR Markers for Genetic Analysis of Phytophthora sojae

作　　者：蔺宇[1] 徐静静[1] 王晓鸣[1] 武小菲[1] 李彦舫[2] 朱振东[1] 

机构地区：[1]中国农业科学院作物科学研究所,国家农作物基因资源与基因改良重大科学工程,北京100081 [2]吉林大学农学院,长春130062

出　　处：《中国农业科学》2008年第8期2294-2301,共8页Scientia Agricultura Sinica

基　　金：国家重点基础研究发展规划“973”项目(2002CB111406)

摘　　要：【目的】开发大豆疫霉菌SSR标记,为深入了解大豆疫霉菌遗传变异提供理想分析工具。【方法】用SSRIT软件对28197条大豆疫霉菌EST进行SSR搜索,选择含有SSR的合适EST设计、合成引物和PCR扩增。【结果】发现1454条EST含有SSR,其中3个碱基重复基元类型最多,有855个,占鉴定总数的54.3%。设计合成140对引物,用10个大豆疫霉菌菌株基因组DNA进行PCR检测,有111对引物(79.3%)扩增出SSR特征条带。通用性检测表明有33对引物在检测的1个或多个其它疫霉菌或腐霉菌中产生扩增产物。【结论】大豆疫霉菌EST含有丰富的SSR位点,本研究从EST中开发了111个新大豆疫霉菌SSR标记,可有效地用于大豆疫霉菌及其相关种的遗传变异研究。[Objective] This study was aimed at developing SSR markers of Phytophthora sojae in order to provide an ideal tool for better understanding of genetic variation of P. sojae. [ Method ] Using SSRIT （simple sequence repeat identification tool） to search SSRs in 28197 ESTs of P. sojae, suitable ESTs harboring SSR were selected for design of primers, and synthesized primers were used to amplify genomic DNA of selected isolates of P. sojae, other Phytophthora species and Pythium. [Result] There were 1 454 ESTs that were found containing SSR. Out of identified SSRs, 855 （54.3%） were trinucleotide repeats, and they were the most frequent repeats in P. sojae. Among the 140 primer pairs designed and synthesized, there were 111 （79.3%） primer pairs that amplified characteristic SSR bands in genomic DNA of 10 tested isolates of P. sojae. The results for transferability test showed that 33 primer pairs could have amplification products in tested one or more isolates of other Phytophthora species or Pythium sp. [Conclusion] There were abundant SSR loci in EST of P. sojae, in this study, 111 novel EST-SSR markers of P. sojae were developed, and these markers should be useful for genetic variation study of P. sojae and its relative species.

关 键 词：大豆疫霉菌 表达序列标签(EST) SSR标记 通用性 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治] S513[农业科学—植物保护]