National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan.
DNA Res. 2011 Apr;18(2):107-15. doi: 10.1093/dnares/dsr003. Epub 2011 Mar 30.
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.
在常用的分子标记中，简单重复序列（SSR，也称为微卫星）具有在个体位点上具有高水平多态性和共显性遗传模式等优点。为了促进大豆的系统和快速遗传作图，我们设计了一个由 304 个 SSR 标记组成的基因型面板，这些标记是根据等位基因多样性和染色体位置选择的，以提供广泛的覆盖范围。该面板中大多数标记的引物对都经过重新设计，以便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，并使用四种不同荧光染料之一进行标记。每组反应的 6 到 8 对引物的多重 PCR 产生了三个不同作图群体中 304 个 SSR 位点中的 283 个位点的等位基因数据，这些位点与先前确定的位置相同。用四重 PCR 对 87 份不同大豆种质资源的每条染色体上的 4 个 SSR 进行等位基因多样性分析。这 80 个位点的平均等位基因数和多态性信息量值分别为 14.8 和 0.78。SSR 基因型面板的高水平多态性、易于分析和高度准确性应该使其广泛适用于大豆遗传学和育种。
