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利用 Illumina MiSeq 测序技术分离和鉴定 21 个多态性微卫星位点用于岩虾 Palaemon elegans。

Isolation and characterization of 21 polymorphic microsatellite loci for the rockpool shrimp Palaemon elegans using Illumina MiSeq sequencing.

机构信息

Department of Biology, Centro de Investigaciones Científicas Avanzadas (CICA) and Evolutionary Biology Group (GIBE), Universidade da Coruña, Campus da Zapateira, A Coruña, E-15008, Spain.

AllGenetics & Biology SL, Edificio CICA, A Coruña, E-15071, Spain.

出版信息

Sci Rep. 2018 Nov 21;8(1):17197. doi: 10.1038/s41598-018-35408-1.

Abstract

The rockpool shrimp Palaemon elegans is considered an important crustacean species within the European coastline fauna. This species is experiencing an ongoing geographical expansion beyond its native distribution range due to unintentional human introductions. A better knowledge of the genetic diversity, geographic structure and connectivity of its populations is necessary. In the present study, microsatellite loci were isolated using the Illumina MiSeq platform. The microsatellite-enriched library sequencing produced 3.9 million raw reads. Reads were processed and primer pairs were designed for microsatellite sequences amplification. Ninety-six microsatellite loci were preliminary screened in individuals from Atlantic and Mediterranean localities. From them, 21 loci exhibited reliable polymorphism and were thoroughly characterized in 30 individuals from a Cantabrian locality (Spain). No linkage disequilibrium between pairs of loci was detected. Number of alleles per locus ranged from 2 to 12. Observed and expected heterozygosities ranged from 0.033 to 0.833 and from 0.033 to 0.869 respectively. No significant departure from the Hardy-Weinberg equilibrium was detected in most of loci. This is the first time that microsatellite markers have been developed for P. elegans. This characterized microsatellite suite provides new suitable tools for further analyses, facilitating the understanding of population genetics both in natural and introduced populations.

摘要

拟穴青虾(Palaemon elegans)被认为是欧洲沿海动物群中一种重要的甲壳类物种。由于人类的无意引入,该物种正在持续向其原生分布范围以外的地区扩张。因此,有必要更好地了解其种群的遗传多样性、地理结构和连通性。在本研究中,使用 Illumina MiSeq 平台分离了微卫星位点。微卫星富集文库测序产生了 390 万条原始reads。对reads 进行了处理,并为微卫星序列的扩增设计了引物对。初步筛选了来自大西洋和地中海地区个体的 96 个微卫星位点。从中筛选出 21 个具有可靠多态性的位点,并在西班牙坎塔布里亚地区的 30 个个体中对其进行了全面特征分析。未检测到对位点之间的连锁不平衡。每个位点的等位基因数范围从 2 到 12。观察到的和预期的杂合度分别在 0.033 到 0.833 和 0.033 到 0.869 之间。大多数位点未偏离哈迪-温伯格平衡。这是首次为拟穴青虾开发微卫星标记。该特征微卫星套件为进一步的分析提供了新的合适工具,有助于了解自然和引入种群的群体遗传学。

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