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利用简化基因组测序技术进行甜瓜全基因组单核苷酸多态性发现及高密度遗传图谱构建

Genome-Wide Single Nucleotide Polymorphism Discovery and the Construction of a High-Density Genetic Map for Melon ( L.) Using Genotyping-by-Sequencing.

作者信息

Chang Che-Wei, Wang Yu-Hua, Tung Chih-Wei

机构信息

Department of Agronomy, National Taiwan University Taipei, Taiwan.

Crop Science Division, Taiwan Agricultural Research Institute, Council of Agriculture Taichung, Taiwan.

出版信息

Front Plant Sci. 2017 Feb 6;8:125. doi: 10.3389/fpls.2017.00125. eCollection 2017.

Abstract

Although genotyping-by-sequencing (GBS) enables the efficient and low-cost generation of large numbers of markers, the utility of resultant genotypes are limited, because they are enormously error-prone and contain high proportions of missing data. In this study, we generated single nucleotide polymorphism (SNP) markers for 109 recombinant inbred lines of melon ( L.) using the GBS approach and ordered them according to their physical position on the draft double haploid line DHL92 genome. Next, by investigating associations between these SNPs, we discovered that some segments on the physical map conflict with linkage relationships. Therefore, to filter out error-prone loci, 4,110 SNPs in which we have a high degree of confidence were selected as anchors to test independence with respect to unselected markers, and the resultant dataset was then analyzed using the Full-Sib Family Haplotype (FSFHap) algorithm in the software TASSEL 5.2. On the basis of this analysis, 22,933 loci that have an average rate of missing data of 0.281% were used to construct a genetic map, which spans 1,088.3 cM across 12 chromosomes and has a maximum spacing of 6.0 cM. Use of this high-quality linkage map enabled the identification of several quantitative trait loci (QTL) known to control traits in fruit and validated our approach. This study highlights the utility of GBS markers for the identification of trait-associated QTLs in melon and facilitates further investigation of genome structure.

摘要

虽然基于测序的基因分型(GBS)能够高效且低成本地生成大量标记,但所得基因型的效用有限,因为它们极易出错且缺失数据比例很高。在本研究中,我们使用GBS方法为109个甜瓜重组自交系生成了单核苷酸多态性(SNP)标记,并根据它们在双单倍体品系DHL92基因组草图上的物理位置进行排序。接下来,通过研究这些SNP之间的关联,我们发现物理图谱上的一些片段与连锁关系存在冲突。因此,为了筛选出易出错的位点,我们选择了4110个高度可信的SNP作为锚定标记,以测试它们与未选择标记的独立性,然后使用软件TASSEL 5.2中的全同胞家系单倍型(FSFHap)算法对所得数据集进行分析。基于此分析,我们使用平均缺失数据率为0.281%的22933个位点构建了一个遗传图谱,该图谱跨越12条染色体,长度为1088.3厘摩,最大间距为6.0厘摩。使用这个高质量的连锁图谱能够鉴定出几个已知控制果实性状的数量性状位点(QTL),并验证了我们的方法。本研究突出了GBS标记在甜瓜中鉴定性状相关QTL的效用,并有助于进一步研究基因组结构。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9167/5292975/3dea9256770e/fpls-08-00125-g001.jpg

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