小麦穗发芽抗性相关基因Tamyb10-D1特异性分子标记开发

Development of Gene-specific Markers for Tamyb10-D1 Related to Wheat Pre-Harvest Sprouting Resistance

Tamyb10-D1基因与穗发芽抗性相关,已报道的该基因分子标记扩增产物大,对DNA模板质量要求较高,PCR反应耗时长。为了使该基因在小麦分子育种中得到高效和快速利用,本研究设计了多个引物组合,经检测可特异性地扩增3D基因组上的Tamyb10基因,引物组合380s/703a、380s/1182a、652s/1182a、872s/1182a和872s/1419a扩增效果较好,而且含有652s或703a的引物组合可区分小麦属和粗山羊草属的myb10-D1基因型。鉴于这些分子标记为显性标记,通过多重PCR反应,可确保PCR检测结果的准确性。其中用引物组合380s/652s/872s/1182a进行三重PCR检测Tamyb10-D1基因型结果较好。因此,本研究开发的分子标记可应用于Tamyb10-D1基因的基因型检测和分子辅助育种,对于小麦穗发芽抗性育种工作具有一定的应用价值。

Tamyb10-D1 gene was related to wheat pre-harvest sprouting resistance.The molecular markers of Tamyb10-D1 gene had been developed.But the lengths of the amplified fragments by using these markers were too long.Which meant more better quality of DNA templates and long PCR time were needed to amplify long fragments.To promote the application of Tamyb10-D1 gene in wheat molecular breeding programs,several new primers were developed.The specificity of these primers were confirmed that they could be used to amplify Tamyb10 gene in 3D genome.Among them,primer combinations 380s/703a,380s/1182a,652s/1182a,872s/1182a and 872s/1419a were much better.Moreover,the primer combinations containing 652s or 703a could be used to distinguish myb10-D1 genotype between Triticum aestivum and Aegilops tauschii.Multiple PCR were also used to guarantee the accuracy of PCR.Primer combinations 380s/652s/872s/1182a were more efficient to amplify Tamyb-10 D1 gene in multiple PCR.All above results showed that these specific primers of Tamyb10-D1 gene were good enough to be used as marker resources for germplasms identification,genetic diversity and marker-assisted selection breeding in wheat.