Department of Biochemistry, Faculty of Science, Khon Kaen University, Muang, Khon Kaen, 40002, Thailand.
Mol Biol Rep. 2012 May;39(5):5201-10. doi: 10.1007/s11033-011-1317-x. Epub 2011 Dec 18.
Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to investigate the genetic structure of four subpopulations of Mystus nemurus in Thailand. The 7 RAPD and 7 ISSR primers were selected. Of 83 total RAPD fragments, 80 (96.39%) were polymorphic loci, and of 81 total ISSR fragments, 75 (92.59%) were polymorphic loci. Genetic variation and genetic differentiation obtained from RAPD fragments or ISSR fragments showed similar results. Percentage of polymorphic loci (%P), observed number of alleles, effective number of alleles, Nei's gene diversity (H) and Shannon's information index revealed moderate to high level of genetic variations within each M. nemurus subpopulation and overall population. High levels of genetic differentiations were received from pairwise unbiased genetic distance (D) and coefficient of differentiation. Mantel test between D or gene flow and geographical distance showed a low to moderate correlation. Analysis of molecular variance indicated that variations among subpopulations were higher than those within subpopulations. The UPGMA dendrograms, based on RAPD and ISSR, showing the genetic relationship among subpopulations are grouped into three clusters; Songkhla (SK) subpopulation was separated from the other subpopulations. The candidate species-specific and subpopulation-specific RAPD fragments were sequenced and used to design sequence-characterized amplified region primers which distinguished M. nemurus from other species and divided SK subpopulation from the other subpopulations. The markers used in this study should be useful for breeding programs and future aquacultural development of this species in Thailand.
随机扩增多态性 DNA (RAPD) 和简单重复间序列 (ISSR) 标记被用于研究泰国 4 个湄公裂腹鱼亚种群的遗传结构。共选择了 7 个 RAPD 和 7 个 ISSR 引物。在 83 个 RAPD 片段中,80 个(96.39%)为多态性位点,在 81 个 ISSR 片段中,75 个(92.59%)为多态性位点。RAPD 片段和 ISSR 片段获得的遗传变异和遗传分化结果相似。多态性位点百分比(%P)、观察等位基因数、有效等位基因数、Nei 基因多样性(H)和 Shannon 信息指数表明,每个湄公裂腹鱼亚种群和总体种群内均存在中等到高水平的遗传变异。通过无偏基因距离(D)和分化系数获得的遗传分化水平较高。D 或基因流与地理距离之间的 Mantel 检验显示出低度到中度的相关性。基于分子方差分析表明,亚种群间的变异高于亚种群内的变异。基于 RAPD 和 ISSR 的 UPGMA 聚类树状图显示,亚种群间的遗传关系聚类为 3 个群;宋卡(SK)亚种群与其他亚种群分离。对候选种特异性和亚种群特异性 RAPD 片段进行测序,并用于设计序列特征扩增区域引物,可将湄公裂腹鱼与其他物种区分开来,并将 SK 亚种群与其他亚种群区分开来。本研究中使用的标记应有助于该物种在泰国的养殖计划和未来水产养殖发展。