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利用下一代测序技术开发巨脂鲤的微卫星标记

Development of microsatellite markers using next-generation sequencing for the fish Colossoma macropomum.

作者信息

Ariede Raquel B, Freitas Milena V, Hata Milene E, Matrochirico-Filho Vito A, Utsunomia Ricardo, Mendonça Fernando F, Foresti Fausto, Porto-Foresti Fábio, Hashimoto Diogo T

机构信息

Centro de Aquicultura da Unesp, Universidade Estadual Paulista, Jaboticabal, SP, Brazil.

Departamento de Morfologia, IBB, Universidade Estadual Paulista, Botucatu, SP, Brazil.

出版信息

Mol Biol Rep. 2018 Feb;45(1):9-18. doi: 10.1007/s11033-017-4134-z. Epub 2017 Dec 20.

Abstract

Tambaqui (Colossoma macropomum) is a fish species from the Amazon and Orinoco Rivers, with favorable characteristics to the cultivation system and great market acceptance in South America. However, the construction of a genetic map for the genetic improvement of this species is limited by the low number of molecular markers currently described. Thus, this study aimed to validate gene-associated and anonymous (non-genic) microsatellites obtained by next generation sequencing (RNA-seq and whole genome shotgun-WGS, respectively), for future construction of a genetic map and search for quantitative trait loci (QTL) in this species. In the RNA-seq data, the observed and expected heterozygosity (H and H) ranged from 0.09 to 0.73, and 0.09 to 0.85, respectively. In the WGS data, H and H ranged from 0.33 to 0.95, and 0.28 to 0.92, respectively. In general, the evaluation of 200 markers resulted in 45 polymorphic loci, of which 14 were gene-associated (RNA-Seq) and 31 were anonymous (WGS). Moreover, some markers were related to genes of the immune system, biological regulation/control and biogenesis. This study contributes to increase the number of molecular markers available for genetic studies in C. macropomum, which will allow the development of breeding programs assisted by molecular markers.

摘要

坦巴基鱼(巨脂鲤)是一种来自亚马逊河和奥里诺科河的鱼类,具有适合养殖系统的优良特性,在南美洲市场上广受欢迎。然而,目前该物种遗传改良所需的遗传图谱构建受到已描述分子标记数量较少的限制。因此,本研究旨在验证通过下一代测序(分别为RNA测序和全基因组鸟枪法测序-WGS)获得的基因相关微卫星和匿名(非基因)微卫星,以便未来构建该物种的遗传图谱并寻找数量性状位点(QTL)。在RNA测序数据中,观察到的杂合度(H)和预期杂合度(H)分别为0.09至0.73和0.09至0.85。在全基因组鸟枪法测序数据中,H和H分别为0.33至0.95和0.28至0.92。总体而言,对200个标记的评估产生了45个多态性位点,其中14个是基因相关的(RNA测序),31个是匿名的(全基因组鸟枪法测序)。此外,一些标记与免疫系统、生物调节/控制和生物发生的基因有关。本研究有助于增加可用于巨脂鲤遗传研究的分子标记数量,这将有助于开展分子标记辅助育种计划。

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