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快速创建拟南芥双单倍体系用于数量性状位点作图。

Rapid creation of Arabidopsis doubled haploid lines for quantitative trait locus mapping.

机构信息

Department of Plant Biology, Genome Center, and Howard Hughes Medical Institute, University of California, Davis, CA 95616, USA.

出版信息

Proc Natl Acad Sci U S A. 2012 Mar 13;109(11):4227-32. doi: 10.1073/pnas.1117277109. Epub 2012 Feb 27.

Abstract

Quantitative trait loci (QTL) mapping is a powerful tool for investigating the genetic basis of natural variation. QTL can be mapped using a number of different population designs, but recombinant inbred lines (RILs) are among the most effective. Unfortunately, homozygous RIL populations are time consuming to construct, typically requiring at least six generations of selfing starting from a heterozygous F(1). Haploid plants produced from an F(1) combine the two parental genomes and have only one allele at every locus. Converting these sterile haploids into fertile diploids (termed "doubled haploids," DHs) produces immortal homozygous lines in only two steps. Here we describe a unique technique for rapidly creating recombinant doubled haploid populations in Arabidopsis thaliana: centromere-mediated genome elimination. We generated a population of 238 doubled haploid lines that combine two parental genomes and genotyped them by reduced representation Illumina sequencing. The recombination rate and parental allele frequencies in our population are similar to those found in existing RIL sets. We phenotyped this population for traits related to flowering time and for petiole length and successfully mapped QTL controlling each trait. Our work demonstrates that doubled haploid populations offer a rapid, easy alternative to RILs for Arabidopsis genetic analysis.

摘要

数量性状位点(QTL)作图是研究自然变异遗传基础的有力工具。可以使用多种不同的群体设计来定位 QTL,但重组近交系(RIL)是最有效的方法之一。不幸的是,纯合 RIL 群体的构建耗时耗力,通常需要从杂合 F1 开始至少六代自交。来自 F1 的单倍体植物结合了两个亲本基因组,在每个基因座上只有一个等位基因。将这些不育的单倍体转化为可育的二倍体(称为“加倍单倍体”,DH),只需两步即可产生不朽的纯合系。在这里,我们描述了一种在拟南芥中快速创建重组加倍单倍体群体的独特技术:着丝粒介导的基因组消除。我们生成了一个由 238 个加倍单倍体组成的群体,这些单倍体结合了两个亲本基因组,并通过简化代表度 Illumina 测序对它们进行了基因型分析。我们的群体中的重组率和亲本等位基因频率与现有 RIL 群体中的相似。我们对该群体进行了与开花时间和叶柄长度相关的性状的表型分析,并成功地定位了控制每个性状的 QTL。我们的工作表明,加倍单倍体群体为拟南芥遗传分析提供了一种快速、简单的 RIL 替代方法。

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