INRA UBP UMR 1095, Genetics, Diversity & Ecophysiology of Cereals, Clermont Ferrand, France.
Plant Biotechnol J. 2010 Feb;8(2):196-210. doi: 10.1111/j.1467-7652.2009.00477.x.
In wheat, the deployment of marker-assisted selection has long been hampered by the lack of markers compatible with high-throughput cost-effective genotyping techniques. Recently, insertion site-based polymorphism (ISBP) markers have appeared as very powerful new tools for genomics and genetic studies in hexaploid wheat. To demonstrate their possible use in wheat breeding programmes, we assessed their potential to meet the five main requirements for utilization in MAS: flexible and high-throughput detection methods, low quantity and quality of DNA required, low cost per assay, tight link to target loci and high level of polymorphism in breeding material. Toward this aim, we developed a programme, IsbpFinder, for the automated design of ISBP markers and adapted three detection methods (melting curve analysis, SNaPshot Multiplex System and Illumina BeadArray technology) for high throughput and flexible detection of ISBP or ISBP-derived SNP markers. We demonstrate that the high level of polymorphism of the ISBPs combined with cost-effective genotyping methods can be used to efficiently saturate genetic maps, discriminate between elite cultivars, and design tightly linked diagnostic markers for virtually all target loci in the wheat genome. All together, our results suggest that ISBP markers have the potential to lead to a breakthrough in wheat marker-assisted selection.
在小麦中，由于缺乏与高通量、低成本基因分型技术兼容的标记，标记辅助选择的应用一直受到阻碍。最近，基于插入位点多态性（ISBP）的标记作为六倍体小麦中基因组学和遗传研究的非常强大的新工具出现了。为了证明它们在小麦育种计划中的可能用途，我们评估了它们在 MAS 中满足五个主要要求的潜力：灵活和高通量的检测方法、所需 DNA 的数量和质量低、每个检测的成本低、与目标位点紧密连锁以及在育种材料中具有高水平的多态性。为此，我们开发了一个自动设计 ISBP 标记的程序 IsbpFinder，并适应了三种检测方法（熔解曲线分析、SNaPshot 多重系统和 Illumina BeadArray 技术），用于高通量和灵活检测 ISBP 或 ISBP 衍生 SNP 标记。我们证明，ISBPs 的高水平多态性与具有成本效益的基因分型方法相结合，可用于有效地饱和遗传图谱，区分优良品种，并为小麦基因组中的几乎所有目标位点设计紧密连锁的诊断标记。总的来说，我们的结果表明，ISBP 标记有可能在小麦标记辅助选择方面取得突破。
