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基于EST信息的百合SSR标记的建立
引用本文:杨素丽,明军,刘春,穆鼎,李名扬.基于EST信息的百合SSR标记的建立[J].园艺学报,2008,35(7):1069-1074.
作者姓名:杨素丽  明军  刘春  穆鼎  李名扬
作者单位:(1中国农业科学院蔬菜花卉研究所,北京 100081; 2西南大学园艺园林学院,重庆 400715 )
基金项目:国家社会公益研究专项基金,国家高技术研究发展计划(863计划),国家科技支撑计划,北京市花卉重点项目
摘    要:依据已知的百合EST(expressed sequence tags)序列信息,开发新的SSR(simple sequence repeats)标记,在NCBI的EST数据库1 688 EST中检索到98条含有101个SSR的序列, SSR的检出率为5.98%,其中三核苷酸重复类型占主导地位,出现频率为2.84%。EST-SSR的重复基元共搜索到47种,其中三核苷酸重复基元类型最为丰富,大约占重复基元类型总数的一半。利用部分EST-SSRs序列共设计23对SSR引物, 以铁炮百合‘Snow Queen' DNA为模板,对引物进行筛选,其中18对引物有扩增产物,占所设计引物总数的78.26%;进一步用这些引物在5个杂种系列13个百合品种进行多态性测试,显示多态性的引物占可扩增引物的66.7%。本研究结果证明了基于百合EST信息建立SSR标记是一种有效而又可行的方法。

关 键 词:百合  EST  SSR  引物设计  
收稿时间:2008-3-25
修稿时间:2008-5-23

Data Mining for Simple Sequence Repeats Marker Development in Expressed Sequence Tags from Lilium L.
YANG Su-li,MING Jun,LIU Chun,MU Ding,LI Ming-yang.Data Mining for Simple Sequence Repeats Marker Development in Expressed Sequence Tags from Lilium L.[J].Acta Horticulturae Sinica,2008,35(7):1069-1074.
Authors:YANG Su-li  MING Jun  LIU Chun  MU Ding  LI Ming-yang
Institution:(;1.Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;;2.College of Horticulture and Landscape,Southwest University,Chongqing 400715,China)
Abstract:The growing availability of EST sequences from Lilium L. provides a potential valuable source of new SSR markers. In this study, 101 SSR-ESTs from 1688 ESTs in the National Center for Biotechnology Information (NCBI) database, representing 5.98% of the total number of ESTs were identified .Among them, the trinucleotide repeat is the dominant type with repeat motifs being the most common, accounting for 2.84% of ESTs. 47 kinds of repeat motifs were mined out from all SSRs. 23 SSR primers were designed to sequence flanking SSRs, the primer pairs designed were screened against genomic DNA of‘Snow Queen’from which most EST-SSRs were derived, and 18 primer pairs showed the amplification, accounting for 78.26% of total primers. Then the primers showing amplification were subjected to PCR for DNA from 13 Lilium L. cultivars of 5 hybridism series and 12 primer pairs showed polymorphisms, accounting for 66.7% of primers available. Results prove that it is an effective and feasible approach to develop SSR markers based on ESTs in Lilium L.
Keywords:EST  SSR
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