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开发并评估了用于二倍体甜樱桃和四倍体酸樱桃的全基因组 6K SNP 芯片。

Development and evaluation of a genome-wide 6K SNP array for diploid sweet cherry and tetraploid sour cherry.

机构信息

Department of Horticulture, Washington State University, Pullman, Washington, United States of America.

出版信息

PLoS One. 2012;7(12):e48305. doi: 10.1371/journal.pone.0048305. Epub 2012 Dec 20.

Abstract

High-throughput genome scans are important tools for genetic studies and breeding applications. Here, a 6K SNP array for use with the Illumina Infinium® system was developed for diploid sweet cherry (Prunus avium) and allotetraploid sour cherry (P. cerasus). This effort was led by RosBREED, a community initiative to enable marker-assisted breeding for rosaceous crops. Next-generation sequencing in diverse breeding germplasm provided 25 billion basepairs (Gb) of cherry DNA sequence from which were identified genome-wide SNPs for sweet cherry and for the two sour cherry subgenomes derived from sweet cherry (avium subgenome) and P. fruticosa (fruticosa subgenome). Anchoring to the peach genome sequence, recently released by the International Peach Genome Initiative, predicted relative physical locations of the 1.9 million putative SNPs detected, preliminarily filtered to 368,943 SNPs. Further filtering was guided by results of a 144-SNP subset examined with the Illumina GoldenGate® assay on 160 accessions. A 6K Infinium® II array was designed with SNPs evenly spaced genetically across the sweet and sour cherry genomes. SNPs were developed for each sour cherry subgenome by using minor allele frequency in the sour cherry detection panel to enrich for subgenome-specific SNPs followed by targeting to either subgenome according to alleles observed in sweet cherry. The array was evaluated using panels of sweet (n = 269) and sour (n = 330) cherry breeding germplasm. Approximately one third of array SNPs were informative for each crop. A total of 1825 polymorphic SNPs were verified in sweet cherry, 13% of these originally developed for sour cherry. Allele dosage was resolved for 2058 polymorphic SNPs in sour cherry, one third of these being originally developed for sweet cherry. This publicly available genomics resource represents a significant advance in cherry genome-scanning capability that will accelerate marker-locus-trait association discovery, genome structure investigation, and genetic diversity assessment in this diploid-tetraploid crop group.

摘要

高通量基因组扫描是遗传研究和育种应用的重要工具。在这里,我们开发了一种用于 Illumina Infinium®系统的 6K SNP 芯片,用于二倍体甜樱桃(Prunus avium)和四倍体酸樱桃(P. cerasus)。这项工作由 RosBREED 领导,该组织是一个社区倡议,旨在为蔷薇科作物提供标记辅助育种。在多样化的育种种质中进行的下一代测序提供了 250 亿个樱桃 DNA 碱基对,从中鉴定出了甜樱桃和源自甜樱桃(avium 亚基因组)和 P. fruticosa(fruticosa 亚基因组)的两个酸樱桃亚基因组的全基因组 SNP。锚定最近由国际桃基因组计划发布的桃基因组序列,预测了 190 万个假定 SNP 检测到的相对物理位置,初步过滤到 368943 个 SNP。进一步的过滤是根据在 160 个个体上用 Illumina GoldenGate®检测试剂盒检测的 144 个 SNP 子集的结果进行指导的。设计了一个 6K Infinium® II 芯片,用于在甜樱桃和酸樱桃基因组中均匀分布 SNP。通过在酸樱桃检测面板中使用次要等位基因频率来富集亚基因组特异性 SNP,然后根据在甜樱桃中观察到的等位基因靶向到相应的亚基因组,为每个酸樱桃亚基因组开发了 SNP。使用甜樱桃(n = 269)和酸樱桃(n = 330)育种种质的面板对该芯片进行了评估。大约三分之一的芯片 SNP 对每个作物都是有信息的。在甜樱桃中验证了总共 1825 个多态性 SNP,其中 13%是最初为酸樱桃开发的。在酸樱桃中解决了 2058 个多态性 SNP 的等位基因剂量,其中三分之一是最初为甜樱桃开发的。这个可公开获取的基因组资源代表了樱桃基因组扫描能力的重大进展,将加速该二倍体-四倍体作物群体中的标记-基因座-性状关联发现、基因组结构研究和遗传多样性评估。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aff4/3527432/c4e4e6eae31c/pone.0048305.g001.jpg

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