虾夷马粪海胆微卫星标记制备及对3个养殖群体的遗传多样性分析  

Isolation of microsatellite markers and Genetic diversity analysis in 3 cultured populations of sea urchin(Strongylocentrotus intermedius)

作　　者：丁君[1] 李润玲[1] 常亚青[1] 孙效文[1] 

机构地区：[1]大连水产学院农业部海洋水产增养殖学与生物技术重点开放实验室,辽宁大连116023

出　　处：《分子科学学报》2008年第3期173-179,共7页Journal of Molecular Science

基　　金：国家"863"计划项目(2006AA10A411)

摘　　要：通过磁珠富集法分离获得了虾夷马粪海胆微卫星DNA序列160个．其中完美型108个（67．50％），非完美型33个（20．63％），复合型19个（11．88％），微卫星DNA序列最多重复次数达到107次．根据这些微卫星DNA序列的两翼序列，采用Primer5软件设计出虾夷马粪海胆的微卫星引物，通过筛选，采用其中的12对微卫星DNA标记对大连凌水群体（DL）、大连獐子岛群体（DZ）、山东荣城群体（SR）3个虾夷马粪海胆养殖群体的遗传多样性进行分析，共获得73个等位基因，不同引物获得的等位基因数为1．11个，片段大小为84．362bp．除SUX001位点外，其余位点的PIC值在0．2640．0．6705之间，3个群体的平均观测杂合度分别为0．4618（DL），0．4375（SR）和0．4340（DZ），平均期望杂合度分别为0．5026（DL），0．5079（SR）和0．4494（Dz）．Hardy-Weinberg平衡分析显示，73％的被检测位点显著偏离平衡，F-检验显示4个位点的F。值低于0．05，表明群体间存在一定程度的分化．群体间遗传相似性系数、遗传距离及UPGMA聚类分析表明大连獐子岛群体（Dz）与大连凌水群体（DL）群体亲缘关系较近，二者与山东荣成群体（SR）亲缘关系较远．对深入了解我国虾夷马粪海胆养殖群体遗传结构特征及其种质资源状况具有重要意义．In the present study, microsatellites DNA was isolated and enriched from genome of sea urchin populations （Strongylocentrotus intermedius）by magnetic beads. 160 mierosatellite sequence were obtained. The mierosatellite sequences were structurally categorized into follows： perfect 108 （67.50%）, imperfect 33 （20.63%）, ancl compound 19 （ 11.88 % ）. The largest array contained 107 repeats. The microsatellite primers of sea urchins were designed basing on the flanking sequence of the mierosatellite loci with software Primer 5,12 mierosatellite markers were developed and applied to investigate the polymorphism in 3 cultured populations of Strongylocentrotus intermedius,including the population of Lingshui area Of Dalian （DL） ,Rongcheng area of Shandong （SR） and Zhangzidao island area of Dalian （DZ）. Seventy-threc alleles were acquired, with allele： size ranging from 84 to 362 bp. The number of alle- les of each locus ranged from 1 to 11. Except SUX001, the PIC of per locus ranged from0.264 0 to 0.670 5.The average observed heterozygosity （Ho） was 0.461 8（DL）,0.437 5 （SR） and 0.434 0（DZ）,respectively.The average expected heterozygosity （He） was 0. 502 6 （DL）, 0. 507 9 （SR） and 0. 449 4 （DZ）, respectively. The analysis of Hardy-Weinberg equilibrium showed that 73% alleles departed from Hardy-Weinberg equilibrium significantly. The F-test showed that the Fst value of 4 loci was lower than 0.05, that means there is some genetic differentiation in 3 cultured populations of sea urchin at a certain extent. According the results of genetic similarity, genetic distance and UPGMA dendrogram,we found that the population of DL and DZ are closer than those from SR.This research is significance for understanding the genetic structure character and resource status of Strongylocentrotus intennedius in China.

关 键 词：虾夷马粪海胆 微卫星 遗传多样性 

分 类 号：Q785[生物学—分子生物学]