基于山药转录组测序的EST-SSR信息分析及分子标记开发  

EST-SSR Information Analysis and Molecular Marker Development Based on Chinese yam Transcriptome Sequencing

作　　者：李锦超 董俊美 贾凯旋 孟义江[2] 杨太新[1] 侯孟泽 葛淑俊[1] Li Jinchao;Dong Junmei;Jia Kaixuan;Meng Yijiang;Yang Taixin;HouMengze;Ge Shujun(Key Laboratory of Crop Germplasm Resources research and Utilization in North China,Ministry of Education,College of Agronomy,Hebei Agricultural University,Baoding,071000;College of Life Science,Hebei Agricultural University,Baoding,071000)

机构地区：[1]河北农业大学农学院,教育部华北作物种质资源研究与利用重点实验室,保定071000 [2]河北农业大学生命科学学院,保定071000

出　　处：《分子植物育种》2025年第4期1269-1277,共9页Molecular Plant Breeding

基　　金：河北省现代农业产业技术体系中药材产业创新团队项目(HBCT201806202)资助。

摘　　要：山药(Dioscorea opposita Thunb.)为药食同源作物,为解析其遗传背景,本研究基于山药珠芽转录组数据挖掘SSR位点。结果表明,在46701条unigenes中检测到24467个SSR,平均每2.29 kb发现1个SSR位点,发生频率33.78%,出现频率53.40%。重复基元类型中单核苷酸占比52.64%,A/T基元最多,其次,二、三核苷酸重复类型分别占23.66%和20.42%,六核苷酸重复类型最为丰富,有112种;SSR基元重复次数多为5~10次,除单碱基类型外,6次重复位点最多;SSR基序长度主要为10~15 bp,占总数的59.64%。随机合成50对不同重复基元类型EST-SSR引物,对29份不同山药种质资源模板进行PCR扩增及电泳分析,47对引物为有效引物,82%的引物呈现多态性;聚类分析表明,29份资源可大致分为3大类群,分为北方山药、南方参薯及混杂类群。以上结果显示,山药转录组SSR位点数量较多、发生频率较高,并且类型丰富,可为山药种质资源鉴定、遗传多样性分析及分子标记辅助育种等提供基础。Dioscorea opposita Thunb.is a medicinal and edible homologous crop.In order to analyze its genetic background,SSR loci were mined based on the transcriptomic data of bulbil of Chinese yam.The results showed that 24467 SSR markers were detected in 46701 unigenes,with an average of 1 SSR loci per 2.29 kb,and the occurrence frequency was 33.78%and 53.40%.Among the repeat primitives,single nucleotide accounted for 52.64%,and A/T was the largest,followed by binucleotide and trinucleotide,23.66%and 20.42%,respectively.There were 112 hexanucleotide repeat types.The repeat times of SSR elements were mostly 5~10 times.Except for single base type,6 repeat sites were the most.The length of SSR motif was mainly 10~15 bp,accounting for 59.64%of the total.50 pairs of EST-SSR primers with different repeat primitive types were randomly synthesized.PCR amplification and electrophoresis analysis were performed on 29 different Chinese yam germplasm resources templates.47 pairs of primers were effective,and 82%of the primers showed polymorphism.Cluster analysis revealed that the 29 resources could be roughly divided into three groups,including northern Chinese yam,southern Chinese Yam and hybrid group.These results indicated that SSR loci were abundant in types,abundant in number and high in frequency,which could provide a basis for germplasm resource identification,genetic diversity analysis and molecular marker-assisted breeding of Chinese yam.

关 键 词：山药 转录组 SSR分子标记 

分 类 号：S632.1[农业科学—蔬菜学]