文献类型：期刊文献

中文题名：基于干旱胁迫下杉木转录组序列的EST-SSR分子标记开发

英文题名：Development of EST-SSR Molecular Marker Based on Sequences of Cunninghami lanceolata Transcripts under Drought Stress

作者：吴夏雷[1] 董黎[1] 孙宇涵[1] 胡瑞阳[2] 郑会全[3] 胡德活[3] 李云[1]

第一作者：吴夏雷

机构：[1]北京林业大学;[2]中国林业科学研究院华北林业实验中心;[3]广东省林业科学研究院

年份：2018

卷号：46

期号：2

起止页码：1-5

中文期刊名：东北林业大学学报

外文期刊名：Journal of Northeast Forestry University

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD_E2017_2018】；

基金：广东省科技计划项目(20168020201002);国家“863”重点项目(2011AA100203);中央高校基本科研业务费专项资金项目(2015ZCQ-SW-03);国家林业局重点项目(2012-06);国家自然科学基金项目(31400562).

语种：中文

中文关键词：杉木转录组;EST-SSR标记;毛细管电泳;多态性

外文关键词：Cunninghami lanceolata transcripts; EST-SSR markers; Capillary electrophoresis; Polymorphism

分类号：S513

摘要：利用干旱胁迫下杉木转录组数据进行杉木EST-SSR分子标记的开发。对干旱胁迫下的杉木组培材料在Solexa mRNA-Seq平台的二代高通量测序技术下进行了转录组测定,序列拼接后共得到75 357个片段,总长度65.29 Mb,共含有EST-SSR位点2 383个,其中三核苷酸重复类型数量最多,占总数的41.21%。随机挑选120个EST-SSR,用Primer Premier 5软件设计引物,再以遗传距离较远的8份杉木DNA样品为模板,通过8%非变性聚丙烯酰胺凝胶电泳,从120对引物中初选出电泳条带较好的24对引物,用24份杉木DNA样品进行毛细管电泳检测。结果表明:在初选出的24对EST-SSR引物中有23对引物检测出特异性峰;在24份杉木样品中共检测到等位基因94个,平均每个位点4.087 0个,等位基因变异数范围2~9个;有效等位基因数范围1.086 8~6.914 3个,平均有效等位基因2.089 0个;观测杂合度范围0.250 0~1.000 0;期望杂合度范围0.082 4~0.896 1;多态性信息指数范围0.173 2~2.035 3。开发得到的一组杉木EST-SSR分子标记为研究杉木的遗传多样性、种群结构、DNA指纹数据库构建和遗传信息的保存提供了基础。
The EST-SSR molecular markers of Cunninghami lanceolata were developed by using the data of Chinese fir transcripts under drought stress. The transgenic plants under drought stress were tested on the Solexa mRNA-Seq platform with the second generation high-throughput sequencing technique. The 75 357 contigs were obtained after splicing, the total length was 65.29 Mb. And there were 2 383 EST-SSR loci, the number of trinucleotide repeats was the largest, accounting for 41.21% of the total. The 120 EST-SSRs were randomly selected and primers were designed by Primer Premier 5. Then, eight C. lanceolata DNA samples were randomly selected as a template and 24 pairs of primers （from 120 pairs of primers） were selected by 8% non-denaturing polyacrylamide gel electrophoresis. The 24 samples of C. lanceolata were used for cap- illary electrophoresis. The result showed 23 pairs of EST-SSR primers were detected with specific peaks. The 94 alleles were detected in 24 samples of C. lanceolata, each loci had an average of 4.087 0. The number of alleles （Na） ranged from 2 to 9. The number of effective alleles （Ne） ranged from 1.086 8 to 6.914 3, and the number of effective allele was 2.089 0. Observed heterozygosity （Ho） ranged from 0.250 0 to 1.000 0. Expected heterozygosity （He） ranged from 0.0824 to 0.896 1. Shannon index ranged from 0.173 2 to 2.035 3. The EST-SSR molecular marker of C. lanceolata provides the basis for the study on genetic diversity, population structure, DNA fingerprint database construction, and genetic information preserva- tion of C. lanceolata.