Development and Application of a Whole-Genome Simple Sequence Repeat Panel for High-Throughput Genotyping in Soybean
Development and Application of a Whole-Genome Simple Sequence Repeat Panel for High-Throughput Genotyping in Soybean
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DOI：
10.1093/dnares/dsr003
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发表时间：
2011-04-01
期刊：
影响因子：
4.1
通讯作者：
Ishimoto, Masao
中科院分区：
文献类型：
--
作者：
Sayama, Takashi;Hwang, Tae-Young;Ishimoto, Masao
在常用的分子标记中，简单重复序列（SSRs，或微卫星）具有较高的多态性和单个位点的共显性遗传模式等优势。为了实现大豆系统、快速的遗传定位，我们设计了一个由304个SSR标记组成的基因分型小组，选择了等位基因多样性和染色体定位，以提供广泛的覆盖范围。面板上大多数标记的引物对经过重新设计，在多重聚合酶链反应（PCR）和荧光测序仪分析中产生80-600 bp的扩增子，并用四种不同的荧光染料中的一种进行标记。在三个不同的定位群体中，每组6 - 8对引物进行多重PCR，得到304个SSR位点中的283个位点的等位基因数据，这些位点的定位与之前确定的位置相同。采用四重PCR方法对87份大豆种质材料的等位基因多样性进行了分析。这80个位点的平均等位基因数和多态性信息含量值分别为14.8和0.78。SSR基因分型面板具有多态性高、易于分析和准确性高等特点，在大豆遗传育种中具有广泛的应用前景。
Among commonly applied molecular markers, simple sequence repeats (SSRs, or microsatellites) possess advantages such as a high level of polymorphism and codominant pattern of inheritance at individual loci. To facilitate systematic and rapid genetic mapping in soybean, we designed a genotyping panel comprised 304 SSR markers selected for allelic diversity and chromosomal location so as to provide wide coverage. Most primer pairs for the markers in the panel were redesigned to yield amplicons of 80-600 bp in multiplex polymerase chain reaction (PCR) and fluorescence-based sequencer analysis, and they were labelled with one of four different fluorescent dyes. Multiplex PCR with sets of six to eight primer pairs per reaction generated allelic data for 283 of the 304 SSR loci in three different mapping populations, with the loci mapping to the same positions as previously determined. Four SSRs on each chromosome were analysed for allelic diversity in 87 diverse soybean germplasms with four-plex PCR. These 80 loci showed an average allele number and polymorphic information content value of 14.8 and 0.78, respectively. The high level of polymorphism, ease of analysis, and high accuracy of the SSR genotyping panel should render it widely applicable to soybean genetics and breeding.