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A resource of large-scale molecular markers for monitoring Agropyron cristatum chromatin introgression in wheat background based on transcriptome sequences

机译:基于转录组序列的小麦背景下土壤冰草染色质基因渗入的大规模分子标记资源

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摘要

Agropyron cristatum is a wild grass of the tribe Triticeae and serves as a gene donor for wheat improvement. However, very few markers can be used to monitor A. cristatum chromatin introgressions in wheat. Here, we reported a resource of large-scale molecular markers for tracking alien introgressions in wheat based on transcriptome sequences. By aligning A. cristatum unigenes with the Chinese Spring reference genome sequences, we designed 9602 A. cristatum expressed sequence tag-sequence-tagged site (EST-STS) markers for PCR amplification and experimental screening. As a result, 6063 polymorphic EST-STS markers were specific for the A. cristatum P genome in the single-receipt wheat background. A total of 4956 randomly selected polymorphic EST-STS markers were further tested in eight wheat variety backgrounds, and 3070 markers displaying stable and polymorphic amplification were validated. These markers covered more than 98% of the A. cristatum genome, and the marker distribution density was approximately 1.28 cM. An application case of all EST-STS markers was validated on the A. cristatum 6 P chromosome. These markers were successfully applied in the tracking of alien A. cristatum chromatin. Altogether, this study provided a universal method of large-scale molecular marker development to monitor wild relative chromatin in wheat.
机译:Agropyron cristatum是Triticeae部落的野草,是小麦改良的基因供体。但是,很少有标记可用于监测小麦中的广ist香染色体染色质渗入。在这里,我们报道了一个基于转录组序列的大规模分子标记资源,可用于跟踪小麦中的外来基因渗入。通过将cr冬青与中国春季参考基因组序列比对,我们设计了9602 cr冬青表达序列标签序列标记位点(EST-STS)标记,用于PCR扩增和实验筛选。结果,6063多态性EST-STS标记对单收割小麦背景中的A. cristatum P基因组具有特异性。在8个小麦品种背景中进一步测试了4956个随机选择的多态EST-STS标记,并验证了3070个显示稳定和多态扩增的标记。这些标记覆盖了超过98%的C. crristatum基因组,标记分布密度约为1.281.cM。所有EST-STS标记的应用案例均已在cristatum 6 P染色体上得到验证。这些标记已成功应用于跟踪外来的A. cristatum染色质。总之,本研究提供了一种大规模分子标记开发的通用方法,以监测小麦中的野生相对染色质。

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