Gennaro Andrea, Koebner Robert M D, Ceoloni Carla
Department of Agrobiology and Agrochemistry, University of Tuscia, via S. Camillo de Lellis, Viterbo, Italy.
Funct Integr Genomics. 2009 Aug;9(3):325-34. doi: 10.1007/s10142-009-0115-1. Epub 2009 Feb 28.
Lr19, one of the few widely effective genes conferring resistance to leaf rust in wheat, was transferred from the wild relative Thinopyrum ponticum to durum wheat. Since Lr19 confers a hypersensitive response to the pathogen, it was considered likely that the gene would be a member of the major nucleotide-binding site (NBS)-leucine-rich repeat (LRR) plant R gene family. NBS profiling, based on PCR amplification of conserved NBS motifs, was applied to durum wheat-Th. ponticum recombinant lines involving different segments of the alien 7AgL chromosome arm, carrying or lacking Lr19. Differential PCR products were isolated and sequenced. From one such sequence (AG15), tightly linked to Lr19, a 4,121-bp full-length cDNA was obtained. Its deduced 1,258 amino acid sequence has the characteristic NBS-LRR domains of plant R gene products and includes a coiled-coil (CC) region typical of monocots. The genomic DNA sequence showed the presence of two exons and a short intron upstream of the predicted stop codon. Homology searches revealed considerable identity of AG15 with the cloned wheat resistance gene Pm3a and a lower similarity with wheat Lr1, Lr21, and Lr10. Quantitative PCR on leaf-rust-infected and non-infected Lr19 carriers proved AG15 to be constitutively expressed, as is common for R genes.
Lr19是少数几个能广泛有效赋予小麦抗叶锈病能力的基因之一,它是从野生近缘种彭提卡偃麦草转移到硬粒小麦中的。由于Lr19对病原体产生过敏反应,因此该基因很可能是主要的核苷酸结合位点(NBS)-富含亮氨酸重复序列(LRR)植物R基因家族的成员。基于保守NBS基序PCR扩增的NBS分析被应用于硬粒小麦-彭提卡偃麦草重组系,这些重组系涉及携带或缺失Lr19的异源7AgL染色体臂的不同区段。分离并测序了差异PCR产物。从一个与Lr19紧密连锁的序列(AG15)中获得了一个4121bp的全长cDNA。其推导的1258个氨基酸序列具有植物R基因产物的特征性NBS-LRR结构域,并包括单子叶植物典型的卷曲螺旋(CC)区域。基因组DNA序列显示在预测的终止密码子上游存在两个外显子和一个短内含子。同源性搜索发现AG15与克隆的小麦抗性基因Pm3a有相当高的一致性,与小麦Lr1、Lr21和Lr10的相似性较低。对感染叶锈病和未感染叶锈病的Lr19携带者进行定量PCR证明,AG15如R基因常见的那样组成型表达。