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利用 SLAF-seq 技术开发长穗偃麦草 7E 染色体特异性分子标记。

The development of 7E chromosome-specific molecular markers for Thinopyrum elongatum based on SLAF-seq technology.

机构信息

College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, China.

出版信息

PLoS One. 2013 Jun 10;8(6):e65122. doi: 10.1371/journal.pone.0065122. Print 2013.

Abstract

Thinopyrum elongatum is an important relative of wheat, it is favored by many researchers for the disease resistant genes that exist in its E genome. Some studies have showed that the 7E chromosome of Th. elongatum contains resistance genes related to Fusarium head blight and wheat rust. Therefore, developing 7E chromosome-specific molecular markers linked to resistance genes will provide an important tool for exploring and using the resistant genes of Th. elongatum. In addition, it would greatly contribute in the effort to cultivate disease-resistant wheat varieties. Featured in high throughput, high-accuracy and low-cost, SLAF-seq technology has been widely used in molecular breeding, system evolution, and germplasm resource detection. Based on SLAF-seq, 518 specific fragments on the 7E chromosome of Th. elongatum were successfully amplified. A total of 135 primers were designed according to 135 randomly selected fragments, and 89 specific molecular markers of Th. elongatum were developed, with efficiencies up to 65.9%. These markers were all detected in a variety of materials, and they are all proved to be specific and stable. These markers can be used not only for detecting the 7E chromosome of Th. elongatum but also for providing an important theoretical and practical basis for wheat breeding by marker-assisted selection (MAS). This paper reports the first application of SLAF-seq technology with a high success rate in developing specific molecular markers for Th. elongatum, providing a strong case for the application of this new technology.

摘要

长穗偃麦草是小麦的重要近缘植物,其 E 基因组中存在抗病基因,受到许多研究者的青睐。一些研究表明,长穗偃麦草的 7E 染色体含有与赤霉病和小麦锈病相关的抗病基因。因此,开发与抗病基因连锁的 7E 染色体特异性分子标记将为探索和利用长穗偃麦草的抗病基因提供重要工具。此外,这将极大地有助于培育抗病小麦品种。SLAF-seq 技术具有高通量、高精度和低成本的特点,已广泛应用于分子育种、系统进化和种质资源检测等领域。基于 SLAF-seq,成功扩增了长穗偃麦草 7E 染色体上的 518 个特异性片段。根据 135 个随机选择的片段共设计了 135 个引物,开发了 89 个长穗偃麦草的特异性分子标记,效率高达 65.9%。这些标记均在多种材料中进行了检测,均表现出特异性和稳定性。这些标记不仅可用于检测长穗偃麦草的 7E 染色体,还可为小麦的标记辅助选择(MAS)育种提供重要的理论和实践基础。本文首次报道了 SLAF-seq 技术在长穗偃麦草特异性分子标记开发方面的高成功率应用,为这项新技术的应用提供了有力证据。

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