王学勇;周晓丽;高伟;崔光红;黄璐琦;刘春生;

• 1:北京中医药大学中药学院
• 2:首都医科大学中医药学院
• 3:中国中医科学院中药研究所

摘要(Abstract)：

目的:建立新近较高覆盖度丹参EST-SSR分子标记,为不同产地来源丹参居群的遗传变异分析奠定基础。方法:对截止2010年11月Genbank下载的丹参EST序列结合HMPL实验室获取的共计1 408条EST序列,进行处理,获取高质量EST序列;经SSRIT软件搜索、分析EST序列中SSR位点的分布规律及类型;在此基础上,运用Websat软件设计EST-SSR引物,经对不同产地丹参样本DNA模板的PCR筛选和条件优化,建立最新EST-SSR分子标记。结果:丹参的EST-SSR种类丰富、覆盖度较大,平均每5.8 kb就检出1条SSR。各种类型出现的频率相差很大,主要重复类型为二核苷酸、三核苷酸重复,分别占SSR总数的63.0%,35.5%,其次为四核苷酸和五核苷酸重复。在重复基元类型中,二核苷酸重复以CT/AG为主,三核苷酸重复以GAA/TCC为主。所优选的36对引物中有29对引物有扩增产物,有效率达80.5%,所选引物对不同产地丹参居群的分子标记出现了多态性。结论:建立的丹参最新EST-SSR分子标记具有较高的覆盖度和多态性,可用于不同产地丹参居群的遗传变异分析。

关键词(KeyWords)： 丹参;EST-SSR;分子标记

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(30801517);; 北京市自然科学基金项目(7092056)

作者(Author): 王学勇;周晓丽;高伟;崔光红;黄璐琦;刘春生;

Email:

参考文献(References)：

• [1]王学勇,崔光红,黄璐琦,等.丹参4-(5′-二磷酸胞苷)-2-C2甲基-D-赤藓醇激酶的cDNA全长克隆及其诱导表达分析[J].药学学报,2008,43(12):1251.
• [2]Wei Gao,Matthew L Hillwig,Luqi Huang,et al.A functionalgenomics approach to tanshinone biosynthesis provides stereo-chemical insights[J].Org Lett,2009,11(22):5170.
• [3]李小白,崔海瑞,张明龙.EST分子标记开发及在比较基因组学中的应用[J].生物多样性,2006,14(6):541.
• [4]Ulrich K,Posselt,Philippe B,et al.Comparative analysis of ge-netic similarity between perennial ryegrass genotypes investigatedwith AFLPs,ISSRs,RAPDs and SSRs[J].Czech J Gene PlantBreed,2006,42(3):87.
• [5]Eujayl I,Sorrells M,Baum M,et al.Assessment of genotypicvariation among cultivated durum wheat based on EST-SSR andgenomic SSRs[J].Euphytica,2001,119:39.
• [6]Cordeiro G M,Casu R,McIntyre C L,et al.Microsatellitemarkers from sugarcane(Saccharumspp.)ESTs cross transfera-ble toErianthusandSorghum[J].Plant Sci,2001,160(6):1115.
• [7]Kantety V R,Rota M L,Matthews E D,et al.Data mining forsimple sequence repeats in expressed sequence tags from barley,maize,rice,sorghum and wheat[J].Plant Mol Biol,2002,48(5/6):501.
• [8]杨成君,王军.人参EST资源的SSR信息分析[J].植物生理学通讯,2008,44(1):69.
• [9]邓科君,张勇,熊丙全,等.药用植物丹参EST-SSR标记的鉴定[J].药学学报,2009,44(10):1165.
• [10]S Senthilvel,B Jayashree,V Mahalakshmi,et al.Developmentand mapping of simple sequence repeat markers for pearl milletfrom data mining of expressed sequence tags[J].BMC Plant Bi-ology,2008,8:119.
• [11]陈相艳,李伟,戴海英,等.大豆EST资源的SSR信息分析[J].大豆科学,2009,28(3):394.
• [12]Cordeiro G M,Casu R,Mclntyre C L,et al.Microsatellite mark-ers fromsugarcane(Saccharumssp.)ESTs cr oss transferable toerianthus and s orghum[J].Plant Sci,2001,160:11152.
• [13]Kantety R V,Rota M L,Matthews D E,et al.Data mining forsimple sequence repeats in exp ressed sequence tags fr ombarley,maize,rice,sorghum and wheat[J].Plant Mol Biol,2002,48:501.
• [14]Gup ta P K,Rustgi S,Sharma S,et al.Transferable EST2 SSRsmarkers for the study of polymorphis m and genetic diversity inbread wheat[J].Mol Genet Genomics,2003,270:315.
• [15]Cardle L,Ramsay L,Milbourne D,et al.Computational and ex-perimental characterization of physically clustered simple se-quence repeats in plants[J],Genetics,2000,156:847.
• [16]Kota R,Varshney R K,Thiel T,et al.Generation and compari-son of EST-derived SSRs and SNPs in Barley(Hordeum vulgareL.)[J].Hereditas,2001,135(2/3):145.
• [17]姜春芽,廖娇,徐小彪,等.植物EST-SSR技术及其应用[J].分子植物育种,2009,7(1):125.
• [18]Powell W,Machray G C,Provan J.Polymorphism revealed bysimple sequence repeats,trends in plant[J].Science,1996,1(7):215.
• [19]Gupta P K,Balyan HS,S harma P C,et al.Microsatellites inplants:a new class of molecular markers[J].Curr Sci,1996,70(1):45.
• [20]Wang C B,Guo WZ,Cai C P,et al.Characterization,develop-ment and exploitation of EST-derived microsatellites inGossypiumraimondiiUlbrich[J].Chin Sci Bull,2006,51(3):316.