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Characterization of the Soybean Genome Using EST-derived Microsatellite Markers.

机译:使用EST衍生的微卫星标记表征大豆基因组。

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We generated a high-density genetic linkage map of soybean using expressed sequence tag (EST)-derived microsatellite markers. A total of 6920 primer pairs (10.9%) were designed to amplify simple sequence repeats (SSRs) from 63 676 publicly available non-redundant soybean ESTs. The polymorphism of two parent plants, the Japanese cultivar 'Misuzudaizu' and the Chinese line 'Moshidou Gong 503', were examined using 10% polyacrylamide gel electrophoresis. Primer pairs showing polymorphism were then used for genotyping 94 recombinant inbred lines (RILs) derived from a cross between the parents. In addition to previously reported markers, 680 EST-derived microsatellite markers were selected and subjected to linkage analysis. As a result, 935 marker loci were mapped successfully onto 20 linkage groups, which totaled 2700.3 cM in length; 693 loci were detected using the 668 EST-derived microsatellite markers developed in this study, the other 242 loci were detected with 105 RFLP markers, 136 genome-derived microsatellite markers, and one phenotypic marker. We examined allelic variation among 23 soybean cultivars/lines and a wild soybean line using 668 mapped EST-derived microsatellite markers (corresponding to 686 marker loci), in order to determine the transferability of the markers among soybean germplasms. A limited degree of macrosynteny was observed at the segmental level between the genomes of soybean and the model legume Lotus japonicus, which suggests that considerable genome shuffling occurred after separation of the species and during establishment of the paleopolyploid soybean genome.
机译:我们使用表达的序列标签(EST)衍生的微卫星标记物生成了大豆的高密度遗传连锁图谱。共设计了6920对引物(10.9%),以扩增来自63 676种公开可获得的非冗余大豆EST的简单序列重复(SSR)。使用10%聚丙烯酰胺凝胶电泳检测了两个亲本植物的日本品种'Misuzudaizu'和中国品系'Moshidou Gong 503'的多态性。然后将显示多态性的引物对用于对源自亲本间杂交的94个重组自交系(RIL)进行基因分型。除了先前报道的标记外,还选择了680个EST衍生的微卫星标记并进行了连锁分析。结果,成功地将935个标记基因座定位到20个连锁组上,总长度为2700.3 cM;使用本研究开发的668个EST衍生的微卫星标记物检测到693个基因座,使用105个RFLP标记,136个基因组衍生的微卫星标记物和一个表型标记物检测了其他242个基因座。为了确定标记在大豆种质间的可转移性,我们使用668个映射的EST衍生的微卫星标记(对应于686个标记位点)检查了23个大豆品种/品系和野生大豆品系之间的等位基因变异。在大豆基因组和豆科植物日本莲的基因组之间的片段水平上观察到有限程度的大同义,这表明在物种分离后和古多倍体大豆基因组建立期间发生了相当大的基因组改组。

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