免疫相关EST-SSRs标记筛选及其在大菱鲆异源雌核发育子代遗传分析中的应用

免疫相关EST-SSRs标记筛选及其在大菱鲆异源雌核发育子代遗传分析中的应用 Application of Immune Related EST-SSR Markers in Hereditary Analysis on Gynogentic Filial Generation of Turbot Scophthalmus maximus 刘滨 LIU Bin1 1 first-author 雷霁霖 LEI Jilin1 1 刘新富 LIU Xinfu1 1 孟振 MENG Zhen1 1 丁福红 DING Fuhong1 1 李振静 LI Zhenjing2 2 中国水产科学研究院黄海水产研究所/青岛市海水鱼类种子工程与生物技术重点实验室 中国水产科学研究院黄海水产研究所/青岛市海水鱼类种子工程与生物技术重点实验室 Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences/Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences/Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology 中国海洋大学海洋生命学院 中国海洋大学海洋生命学院 College of Marine Life Science,Ocean University of China College of Marine Life Science,Ocean University of China 本研究对大菱鲆EST数据库中12 434条序列进行微卫星搜索,共搜索出831条EST-SSR序列,其中l6条为明确标注免疫相关功能的基因序列.根据此l6条EST-SSR序列设计引物,经过PCR扩增检测和反应条件的优化,最终筛选出9对扩增效果理想的EST-SSR标记对大菱鲆减数分裂雌核发育二倍体子代进行遗传变异检测,结果表明:在Scoph-2-1、Scoph-7-1、Scoph-10-1微卫星位点上大菱鲆亲本和雌核发育二倍体子代均为纯合基因型,其余6个微卫星位点（Psetta-1-1、Psetta-1-2、Psetta-3-1、Psetta-3-2、Scoph-4-1、Scoph-6-1）在大菱鲆雌核发育二倍体仔鱼群体中的观测杂合度≥0.700,期望杂合度>0.460,平均期望杂合度为0.503.大菱鲆减数分裂雌核发育二倍体还显示出较高的重组率,平均重组率为89%;综合上述结果推断,9个免疫相关EST-SSRs标记可有效地用于分析大菱鲆减数分裂雌核发育二倍体子代的遗传变异;6个位点的重组率计算结果能够为今后对大菱鲆的着丝粒进行精确定位提供依据. Total 12 434 EST sequences of the Scophthatmus maximus dbEST from NCBI database were analyzed in this research,and finally 831 EST-SSRs were mined out,in which 16 EST-SSRs were annotated of immune related function.Primers were designed based on the 16 EST-SSR sequences.After examination of pcr amplification and condition optimization,9 ideal locus were eventually screened.Analysis of genetic variation in Scophthatmus maximus meiogynogenetic diploid offspring was performed by the use of the screened 9 immune related primers.The result showed that maternal and filial genotypes were all homozygosis in three loci（Scoph-2-1,Scoph-7-1,Scoph-10-1）,and no genotypes were all homozygosis in 6 loci（Psetta-1-1,Psetta-1-2,Psetta-3-1,Psetta-3-2,Scoph-4-1,Scoph-6-1）.The observed heterozygosity and expected heterozygosity in the 6 loci were more than 0.700 and 0.460 respectively,and the mean expected heterozygosity was 0.503.High recombinant ratio also was showed and the mean recombinant ratio was 89%.The result of this research demonstrated that Scophthatmus maximus meiogynogenetic diploids had high recombination rate,and the nine immune related SSR marks could be effectively used to analyze genetic variation in Scophthatmus maximus meiogynogenetic diploid offspring,while the calculated result of recombination rate in the six loci could provide the basis for centromere location of Scophthatmus maximus. 大菱鲆 EST-SSRs 雌核发育 遗传分析 Scophthatmus maximus EST-SSRs gynogenesis hereditary analysis 10.14188/j.1671-8836.2010.03.020 S917.4 农业部农业公益性行业科研专项经费项目（nyhyzx07-046）;现代农业产业技术体系建设专项资金共同资助项目 2010-03-01 2021-04-01 3 56