Guangdong Academy of Agricultural Sciences, Crops Research Institute, Guangzhou, PR China.
BMC Plant Biol. 2010 Jan 27;10:17. doi: 10.1186/1471-2229-10-17.
The construction of genetic linkage maps for cultivated peanut (Arachis hypogaea L.) has and continues to be an important research goal to facilitate quantitative trait locus (QTL) analysis and gene tagging for use in a marker-assisted selection in breeding. Even though a few maps have been developed, they were constructed using diploid or interspecific tetraploid populations. The most recently published intra-specific map was constructed from the cross of cultivated peanuts, in which only 135 simple sequence repeat (SSR) markers were sparsely populated in 22 linkage groups. The more detailed linkage map with sufficient markers is necessary to be feasible for QTL identification and marker-assisted selection. The objective of this study was to construct a genetic linkage map of cultivated peanut using simple sequence repeat (SSR) markers derived primarily from peanut genomic sequences, expressed sequence tags (ESTs), and by "data mining" sequences released in GenBank.
Three recombinant inbred lines (RILs) populations were constructed from three crosses with one common female parental line Yueyou 13, a high yielding Spanish market type. The four parents were screened with 1044 primer pairs designed to amplify SSRs and 901 primer pairs produced clear PCR products. Of the 901 primer pairs, 146, 124 and 64 primer pairs (markers) were polymorphic in these populations, respectively, and used in genotyping these RIL populations. Individual linkage maps were constructed from each of the three populations and a composite map based on 93 common loci were created using JoinMap. The composite linkage maps consist of 22 composite linkage groups (LG) with 175 SSR markers (including 47 SSRs on the published AA genome maps), representing the 20 chromosomes of A. hypogaea. The total composite map length is 885.4 cM, with an average marker density of 5.8 cM. Segregation distortion in the 3 populations was 23.0%, 13.5% and 7.8% of the markers, respectively. These distorted loci tended to cluster on LG1, LG3, LG4 and LG5. There were only 15 EST-SSR markers mapped due to low polymorphism. By comparison, there were potential synteny, collinear order of some markers and conservation of collinear linkage groups among the maps and with the AA genome but not fully conservative.
A composite linkage map was constructed from three individual mapping populations with 175 SSR markers in 22 composite linkage groups. This composite genetic linkage map is among the first "true" tetraploid peanut maps produced. This map also consists of 47 SSRs that have been used in the published AA genome maps, and could be used in comparative mapping studies. The primers described in this study are PCR-based markers, which are easy to share for genetic mapping in peanuts. All 1044 primer pairs are provided as additional files and the three RIL populations will be made available to public upon request for quantitative trait loci (QTL) analysis and linkage map improvement.
构建栽培花生(Arachis hypogaea L.)的遗传连锁图谱一直是一个重要的研究目标,有助于对数量性状基因座(QTL)进行分析和基因标记,从而应用于育种中的标记辅助选择。尽管已经构建了一些图谱,但它们都是使用二倍体或种间四倍体群体构建的。最近发表的种内图谱是从栽培花生的杂交中构建的,其中只有 135 个简单序列重复(SSR)标记在 22 个连锁群中稀疏分布。需要更详细的具有足够标记的连锁图谱,以便进行 QTL 鉴定和标记辅助选择。本研究的目的是利用主要来自花生基因组序列、表达序列标签(EST)和 GenBank 中发布的“数据挖掘”序列的 SSR 标记构建栽培花生的遗传连锁图谱。
从三个杂交中构建了三个重组自交系(RIL)群体,其中一个共同的雌性亲本是高产西班牙市场型月油 13。用 1044 对设计用于扩增 SSR 的引物和 901 对产生清晰 PCR 产物的引物对这四个亲本进行了筛选。在这三个群体中,有 146、124 和 64 个引物(标记)分别在这些群体中表现出多态性,并用于对这些 RIL 群体进行基因分型。从每个群体中构建了单独的连锁图谱,并使用 JoinMap 基于 93 个共同位点创建了综合图谱。综合连锁图谱由 22 个综合连锁群(LG)组成,包含 175 个 SSR 标记(包括已发表的 AA 基因组图谱上的 47 个 SSR),代表 A. hypogaea 的 20 条染色体。综合图谱总长度为 885.4cM,平均标记密度为 5.8cM。3 个群体中的分离偏倚分别为标记的 23.0%、13.5%和 7.8%。这些偏倚的标记倾向于聚集在 LG1、LG3、LG4 和 LG5 上。由于多态性低,仅映射了 15 个 EST-SSR 标记。相比之下,图谱与 AA 基因组之间存在一些标记的潜在同线性、共线性顺序和共线性连锁群的保守性,但并不完全保守。
从三个含有 175 个 SSR 标记的个体图谱中构建了一个综合连锁图谱,包含 22 个综合连锁群。这张综合遗传连锁图谱是第一张真正的四倍体花生图谱。该图谱还包含 47 个 SSR,已用于已发表的 AA 基因组图谱,可用于比较图谱研究。本研究中描述的引物是基于 PCR 的标记,易于在花生中进行遗传作图共享。所有 1044 对引物都作为附加文件提供,三个 RIL 群体将根据需要提供给公众,以进行数量性状基因座(QTL)分析和连锁图谱改进。