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黄承勤,黄英毅,黄欣,王中铎,郭昱嵩.2020.湖栖鳍虾虎鱼微卫星DNA标记的 开发与群体遗传多样性分析.动物学杂志,55(1):67-76.
湖栖鳍虾虎鱼微卫星DNA标记的 开发与群体遗传多样性分析
Development of Microsatellite DNA Markers and Analysis of Population Genetic Diversity in Gobiopterus lacustris
投稿时间:2019-07-27  修订日期:2020-01-08
DOI:10.13859/j.cjz.202001009
中文关键词:  湖栖鳍虾虎鱼  地理群体  微卫星DNA  遗传多样性
英文关键词:Gobiopterus lacustris  Wild populations  Microsatellites  Genetic diversity
基金项目:国家自然科学基金项目(No. 31201996),广东海洋大学创新强校项目(No. 230419055)
作者单位E-mail
黄承勤 广东海洋大学水产学院南海水产经济动物增养殖广东普通高校重点实验室 湛江 524088 441205816@qq.com 
黄英毅 广东海洋大学水产学院南海水产经济动物增养殖广东普通高校重点实验室 湛江 524088 1074163132@qq.com 
黄欣 广东海洋大学水产学院南海水产经济动物增养殖广东普通高校重点实验室 湛江 524088 597256922@qq.com 
王中铎 广东海洋大学水产学院南海水产经济动物增养殖广东普通高校重点实验室 湛江 524088 aduofa@qq.com 
郭昱嵩 广东海洋大学水产学院南海水产经济动物增养殖广东普通高校重点实验室 湛江 524088 gysrabbit@163.com 
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中文摘要:
      为研究雷州半岛湖栖鳍虾虎鱼(Gobiopterus lacustris)遗传多样性,对湖栖鳍虾虎鱼雌雄性腺进行转录组测序,采用MISA 软件进行微卫星分析,共获得25 452个微卫星标记,其中包含14 708个单碱基重复类型,6 175个2碱基重复类型,3、4、5和6碱基重复类型数目分别为4 223、327、15和4。随机选取50个微卫星位点设计引物,能够扩增出清晰稳定条带的有39对,其中,11个位点表现出不同程度的多态,28个位点呈现单态。利用11个具有多态性的微卫星位点分析湖栖鳍虾虎鱼在廉江市高桥镇红树林保护区、湛江东海岛、雷州市附城镇和雷州市九龙山红树林国家湿地公园4个野生群体中的遗传多样性及遗传结构,11个微卫星位点呈现出不同程度的多态性,等位基因数(Na)2 ~ 15,平均等位基因为(6 ± 3.9)个,有效等位基因数(Ne)、平均观测杂合度(Ho)和期望杂合度(He)分别在1.919 ~ 2.485、0.343 ~ 0.465和0.381 ~ 0.483之间,平均多态信息含量(PIC)为0.348 ~ 0.465。Hardy-Weinber平衡分析显示,4个群体的大部分位点未偏离平衡。在每个群体中进行连锁不平衡分析,有18对位点间显著(P < 0.05)或者极显著(P < 0.01)偏离连锁平衡。遗传分化系数在0.107 ~ 0.216之间,表明4个群体的遗传分化达到了中等水平以上。分子方差分析(AMOVA)显示,湖栖鳍虾虎鱼大部分的差异来自于群体内而非群体间。
英文摘要:
      To understand the genetic diversity of Gobiopterus lacustris in the Leizhou Peninsula, transcriptomic sequencing of male and female gonads was performed. Using MISA software, 25 452 microsatellite markers were identified from the transcriptome data. A total of 14 708 single-base repeat types and 6 175 two-base repeat types were identified, while the numbers of three-, four-, five-, and six-base repeats were 4 223, 327, 15 and 4, respectively. By randomly selecting 50 microsatellite loci to design primers, 39 pairs of clear and stable bands could be amplified. The genetic diversity and genetic structure of G. lacustris were analyzed by microsatellite markers. It was showed that 11 microsatellite loci were polymorphic with different degrees of polymorphism, and the numbers of alleles ranged from 2 to 15, with an average number of 6 ± 3.9 (Table 2). The 11 microsatellite loci were used to analyze the genetic diversity of four populations including Zhanjiang Donghai Island population, Leizhou Jiulong Mountain population, Leizhou Fucheng population and Lianjiang Gaoqiao population. The average number of effective alleles of the four G. lacustris populations ranged from 1.919 to 2.485, and the observed heterozygosity and expected heterozygosity distributed at ranges of 0.343﹣0.465 and 0.381﹣0.483, respectively (Table 2). The average polymorphic information content (PIC) ranged from 0.348 to 0.465 (Table 2). Chi-square tests showed that most loci in the 4 G. lacustris groups did not deviate from Hardy-Weinberg equilibrium (P < 0.05) (Table 3). Linkage disequilibrium analysis was performed in each population and 18 pairs of loci were found to be significantly or very significantly (P < 0.05 or P < 0.01) deviated from linkage equilibrium. The genetic differentiation of the 4 populations reached above average level (0.107﹣0.216) (Table 5). Molecular variance analysis (AMOVA) revealed that genetic variation was mainly derived within the population (Table 6).
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