【摘要】 为解析抗病小麦BJ-1中的抗病基因，本课题组前期将其定位于3A染色体727.722×10~6～727.752×10~6bp物理区间，区间内包含一个抗病基因TaRPP13-3A，在线分析发现其为高表达NLR基因.本研究采用基因克隆、荧光定量表达分析、亚细胞定位、病毒诱导基因沉默和过表达转基因技术，分析了抗病基因TaRPP13-3A的功能.结果显示：(1) TaRPP13-3A基因的cDNA序列全长2 700 bp，与中国春小麦相比，序列在CC、NB-ARC和LRR区域存在核苷酸多态性；(2) TaRPP13-3A:GFP融合蛋白主要在细胞膜上表达；(3) TaRPP13-3A能迅速响应白粉菌侵染，且在BJ-1中的表达量高于感病小麦京411；(4)过表达TaRPP13-3A提高了受体小麦津强5的白粉病抗性；(5)病毒诱导TaRPP13-3A基因沉默后，降低了BJ-1对白粉菌的抗性，证明其与白粉病抗性正相关.上述结果表明，TaRPP13-3A可作为一个新的抗白粉病基因资源在小麦抗病育种工作中发挥作用.更多还原

【Abstract】 To characterize the disease resistance gene in the resistant wheat line BJ-1, the gene was located from 727.722 ×10~6 bp to 727.752 × 10~6 bp physical interval on chromosome 3A previously. This interval contains a resistance gene designated TaRPP13-3A, which was identified as a highly expressed NLR gene through online analysis. In this study, gene cloning,quantitative fluorescence expression analysis, subcellular localization, virus-induced gene silencing(VIGS), and overexpression transgenic techniques were adopted to investigate the function of the TaRPP13-3A gene. The results illustrated that:(1) The full-length cDNA of TaRPP13-3A is 2 700 bp, exhibiting nucleotide polymorphisms in the CC, NB-ARC, and LRR domains compared with Chinese Spring wheat；(2) The TaRPP13-3A:GFP fusion protein was predominantly localized to the cell membrane ；(3) TaRPP13-3A rapidly responded to Blumeria graminis f. sp. tritici(Bgt) infection, with significantly higher expression in BJ-1 than in susceptible wheat Jing 411；(4) The overexpression of TaRPP13-3A enhanced powdery mildew resistance in the susceptible wheat JinQiang5 ；(5) VIGS-mediated silencing of TaRPP13-3A compromised Bgt resistance in BJ-1, demonstrating its positive function during powdery mildew resistance. These findings indicate that TaRPP13-3A can serve as a novel powdery mildew resistance gene resource for breeding programs of wheat disease resistance.更多还原