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优化ddRAD-seq在桃基因组中的性能。

Fine-tuning the performance of ddRAD-seq in the peach genome.

作者信息

Aballay Maximiliano Martín, Aguirre Natalia Cristina, Filippi Carla Valeria, Valentini Gabriel Hugo, Sánchez Gerardo

机构信息

Laboratorio de Biotecnología, Estación Experimental Agropecuaria (EEA) San Pedro, INTA, 2930, San Pedro, Argentina.

Instituto de Agrobiotecnología y Biología Molecular-IABiMo-INTA-CONICET, Instituto de Biotecnología, Centro de Investigaciones en Ciencias Veterinarias y Agronómicas, INTA, 1686, Hurlingham, Argentina.

出版信息

Sci Rep. 2021 Mar 18;11(1):6298. doi: 10.1038/s41598-021-85815-0.

Abstract

The advance of Next Generation Sequencing (NGS) technologies allows high-throughput genotyping at a reasonable cost, although, in the case of peach, this technology has been scarcely developed. To date, only a standard Genotyping by Sequencing approach (GBS), based on a single restriction with ApeKI to reduce genome complexity, has been applied in peach. In this work, we assessed the performance of the double-digest RADseq approach (ddRADseq), by testing 6 double restrictions with the restriction profile generated with ApeKI. The enzyme pair PstI/MboI retained the highest number of loci in concordance with the in silico analysis. Under this condition, the analysis of a diverse germplasm collection (191 peach genotypes) yielded 200,759,000 paired-end (2 × 250 bp) reads that allowed the identification of 113,411 SNP, 13,661 InDel and 2133 SSR. We take advantage of a wide sample set to describe technical scope of the platform. The novel platform presented here represents a useful tool for genomic-based breeding for peach.

摘要

新一代测序(NGS)技术的进步使得以合理成本进行高通量基因分型成为可能,尽管在桃的研究中,这项技术的发展还很有限。迄今为止,在桃的研究中仅应用了一种基于用ApeKI进行单一酶切以降低基因组复杂性的标准测序基因分型方法(GBS)。在本研究中,我们通过测试6种双酶切组合以及ApeKI产生的酶切图谱,评估了双酶切RADseq方法(ddRADseq)的性能。酶切组合PstI/MboI与计算机模拟分析结果一致,保留了最多的位点。在此条件下,对一个多样化的种质资源库(191个桃基因型)进行分析,获得了200,759,000条双端(2×250 bp) reads,可鉴定出113,411个单核苷酸多态性(SNP)、13,661个插入缺失(InDel)和2133个简单序列重复(SSR)。我们利用广泛的样本集来描述该平台的技术范围。本文介绍的新平台是桃基于基因组育种的有用工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/44cd/7973760/b209070d40bd/41598_2021_85815_Fig1_HTML.jpg

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