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在韩国太平洋鲍鱼(杂色鲍(鲍科))中鉴定出的新型多态微卫星基因座在野生和放流群体遗传特征分析中的应用

Application of novel polymorphic microsatellite loci identified in the Korean Pacific Abalone (Haliotis diversicolor supertexta (Haliotidae)) in the genetic characterization of wild and released populations.

作者信息

An Hye Suck, Lee Jang Wook, Hong Seong Wan

机构信息

Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Korea.

Jeju Province Fisheries Research Institute, Jeju 699-814, Korea.

出版信息

Int J Mol Sci. 2012;13(9):10750-10764. doi: 10.3390/ijms130910750. Epub 2012 Aug 28.

Abstract

The small abalone, Haliotis diversicolor supertexta, of the family Haliotidae, is one of the most important species of marine shellfish in eastern Asia. Over the past few decades, this species has drastically declined in Korea. Thus, hatchery-bred seeds have been released into natural coastal areas to compensate for the reduced fishery resources. However, information on the genetic background of the small abalone is scarce. In this study, 20 polymorphic microsatellite DNA markers were identified using next-generation sequencing techniques and used to compare allelic variation between wild and released abalone populations in Korea. Using high-throughput genomic sequencing, a total of 1516 (2.26%; average length of 385 bp) reads containing simple sequence repeats were obtained from 86,011 raw reads. Among the 99 loci screened, 28 amplified successfully, and 20 were polymorphic. When comparing allelic variation between wild and released abalone populations, a total of 243 different alleles were observed, with 18.7 alleles per locus. High genetic diversity (mean heterozygosity = 0.81; mean allelic number = 15.5) was observed in both populations. A statistical analysis of the fixation index (F(ST)) and analysis of molecular variance (AMOVA) indicated limited genetic differences between the two populations (F(ST) = 0.002, p > 0.05). Although no significant reductions in the genetic diversity were found in the released population compared with the wild population (p > 0.05), the genetic diversity parameters revealed that the seeds released for stock abundance had a different genetic composition. These differences are likely a result of hatchery selection and inbreeding. Additionally, all the primer pair sets were effectively amplified in another congeneric species, H. diversicolor diversicolor, indicating that these primers are useful for both abalone species. These microsatellite loci may be valuable for future aquaculture and population genetic studies aimed at developing conservation and management plans for these two abalone species.

摘要

皱纹盘鲍(Haliotis diversicolor supertexta)隶属鲍科,是东亚最重要的海洋贝类物种之一。在过去几十年间,该物种在韩国数量急剧下降。因此,人工繁育的幼苗被投放到自然沿海区域,以补充减少的渔业资源。然而,关于皱纹盘鲍遗传背景的信息却很匮乏。在本研究中,利用下一代测序技术鉴定出20个多态性微卫星DNA标记,并用于比较韩国野生和投放的鲍鱼种群之间的等位基因变异。通过高通量基因组测序,从86,011条原始 reads 中获得了总共1516条(2.26%;平均长度385 bp)包含简单序列重复的 reads。在筛选的99个位点中,28个成功扩增,20个具有多态性。比较野生和投放的鲍鱼种群之间的等位基因变异时,共观察到243个不同的等位基因,每个位点平均有18.7个等位基因。两个种群均观察到高遗传多样性(平均杂合度 = 0.81;平均等位基因数 = 15.5)。对固定指数(F(ST))的统计分析和分子方差分析(AMOVA)表明,两个种群之间的遗传差异有限(F(ST) = 0.002,p > 0.05)。虽然与野生种群相比,投放种群的遗传多样性没有显著降低(p > 0.05),但遗传多样性参数显示,为增加种群数量而投放的幼苗具有不同的遗传组成。这些差异可能是人工繁育选择和近亲繁殖的结果。此外,所有引物对在另一个同属物种九孔鲍(H. diversicolor diversicolor)中均能有效扩增,表明这些引物对两种鲍鱼均有用。这些微卫星位点可能对未来旨在为这两种鲍鱼制定保护和管理计划的水产养殖和种群遗传学研究具有重要价值。

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