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利用特异位点扩增片段测序技术在花椰菜中进行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 Polymorphisms Discovery and High-Density Genetic Map Construction in Cauliflower Using Specific-Locus Amplified Fragment Sequencing.

作者信息

Zhao Zhenqing, Gu Honghui, Sheng Xiaoguang, Yu Huifang, Wang Jiansheng, Huang Long, Wang Dan

机构信息

Institute of Vegetables, Zhejiang Academy of Agricultural Sciences Hangzhou, China.

Biomarker Technologies Corporation Beijing, China.

出版信息

Front Plant Sci. 2016 Mar 21;7:334. doi: 10.3389/fpls.2016.00334. eCollection 2016.

Abstract

Molecular markers and genetic maps play an important role in plant genomics and breeding studies. Cauliflower is an important and distinctive vegetable; however, very few molecular resources have been reported for this species. In this study, a novel, specific-locus amplified fragment (SLAF) sequencing strategy was employed for large-scale single nucleotide polymorphism (SNP) discovery and high-density genetic map construction in a double-haploid, segregating population of cauliflower. A total of 12.47 Gb raw data containing 77.92 M pair-end reads were obtained after processing and 6815 polymorphic SLAFs between the two parents were detected. The average sequencing depths reached 52.66-fold for the female parent and 49.35-fold for the male parent. Subsequently, these polymorphic SLAFs were used to genotype the population and further filtered based on several criteria to construct a genetic linkage map of cauliflower. Finally, 1776 high-quality SLAF markers, including 2741 SNPs, constituted the linkage map with average data integrity of 95.68%. The final map spanned a total genetic length of 890.01 cM with an average marker interval of 0.50 cM, and covered 364.9 Mb of the reference genome. The markers and genetic map developed in this study could provide an important foundation not only for comparative genomics studies within Brassica oleracea species but also for quantitative trait loci identification and molecular breeding of cauliflower.

摘要

分子标记和遗传图谱在植物基因组学和育种研究中发挥着重要作用。花椰菜是一种重要且独特的蔬菜;然而,关于该物种的分子资源报道极少。在本研究中,采用了一种新颖的特异性位点扩增片段(SLAF)测序策略,用于在花椰菜的双单倍体分离群体中大规模发现单核苷酸多态性(SNP)并构建高密度遗传图谱。经过处理后,共获得了12.47 Gb的原始数据,包含7792万对末端读数,并检测到两个亲本之间有6815个多态性SLAF。母本的平均测序深度达到52.66倍,父本为49.35倍。随后,利用这些多态性SLAF对群体进行基因分型,并根据若干标准进一步筛选,以构建花椰菜的遗传连锁图谱。最终,1776个高质量的SLAF标记(包括2741个SNP)构成了连锁图谱,平均数据完整性为95.68%。最终图谱的总遗传长度为890.01 cM,平均标记间隔为0.50 cM,覆盖了参考基因组的364.9 Mb。本研究中开发的标记和遗传图谱不仅可为甘蓝物种内的比较基因组学研究提供重要基础,也可为花椰菜的数量性状位点鉴定和分子育种提供重要基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/355e/4800193/e522224c6a7e/fpls-07-00334-g0001.jpg

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