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首页> 外文会议>International Wheat Genetics Symposium >Map-based isolation of the leaf rust disease resistance gene LrlO from the hexaploid wheat (Triticum aestivum L.) genome
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Map-based isolation of the leaf rust disease resistance gene LrlO from the hexaploid wheat (Triticum aestivum L.) genome

机译：从六倍体小麦（Triticum aestivum L.）基因组的叶片锈病抗性基因Lrlo的地图分离

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More than 50 resistance genes (Lr) against the leaf rust fungal pathogen Puccinia triticina have been identified in the wheat gene pool and a large number of them has been extensively used in breeding. Similar to many other agronomically important genes in wheat, Lr genes are only characterised by their phenotype and sometimes by their chromosomal location and map position. For many years the problems of molecular work in the large (1.6 x lO1"' bp), highly repetitive (80%) and hexaploid bread wheat (Triticum aestivum L.) genome have hampered positional cloning. To isolate the LrlO resistance gene, we have used a combination of subgenome map-based cloning and haplotype studies in the genus Triticum. A T. monococcum BAC contig spanning more than 450 kb of the region orthologous to the LrlO locus in hexaploid wheat was established and full-length sequencing of 211 kb spanning the gene identified two resistance gene analogs, rgal and rga2, cosegregating with LrlO in more than 3000 F2 plants. Haplotypestudies at the LrlO locus showed that the two genes are the only promising candidates for LrlO. Their orthologs (TlOrgal and T10rga2-1A) were isolated from the resistant hexaploid wheat variety ThatcherZ.r/0. Evidence for the identity of the gene was obtained through mutational and transformation analysis. Single point mutations were identified in the TlOrgal gene in three independent EMS mutants of ThatcherZ/70 and TlOrgal conferred resistance to leaf rust in transgenic wheat plants demonstrating thatit is the LrlO gene. LrlO encodes a CC-NBS-LRR type of protein with an N-terminal domain that is under diversifying selection. When overexpressed in transgenic wheat plants, LrlO confers enhanced resistance to leaf rust. LrlO has similarities to RPM1 inA. thaliana and to resistance gene analogs in rice and barley but is not closely related to other wheat leaf rust resistance genes based on Southern analysis. We conclude that map-based cloning of genes of agronomic importance in hexaploid wheat is now feasible, opening new perspectives for molecular bread wheat improvement via transgenic strategies and diagnostic allele detection.
机译：在小麦基因库中鉴定了超过50个抗叶片锈病病原体蛋白蛋白杀菌蛋白的抗性基因（LR），其中大量被广泛用于繁殖。与小麦中的许多其他农学上重要基因类似，LR基因仅特征，其表型，有时仅通过它们的染色体位置和地图位置。多年来，大型分子工作的问题（1.6×101）'Bp），高度重复（80％）和六倍倍醇面包小麦（Triticum aestivum L.）基因组具有阻碍的位置克隆。孤立LRLO抗性基因，我们使用基于小属的基于地图的克隆和单倍型研究的组合。建立了跨越450kb的横向于450kb的六倍倍体小麦的450kb型区域的T. monococcum bac contig和全长测序为211kb跨越基因鉴定了两种抗性基因类似物，RGAL和RGA2，在3000多个F2植物中用LR1含有CoSegregating。LRLO基因座的Haplotypesesties表明，这两个基因是LRLO的唯一有希望的候选者。它们的矫形器（Torgal和T10RGA2-1a）是唯一有希望的候选者从抗性六倍体小麦品种Thatcherz.r / 0中分离出来。通过突变和转化分析获得基因的身份的证据。在Tlorgal Gen中鉴定单点突变e在三个独立的EMS突变体中，在转基因小麦植物中表现出甲片的叶锈，表现出甲片的抗叶锈病是LRLO基因。 LRLO编码CC-NBS-LRR类型的蛋白质，其具有在不同选择的N末端结构域。当转基因小麦植物中过表达时，LRLO赋予叶锈的增强抗性。 LRLO与RPM1 INA相似。蒂利亚纳和大麦和大麦的抵抗基因类似物，但与基于南方分析的麦片锈蚀基因密切相关。我们得出结论，基于地图的六倍体小麦农艺意义基因的克隆现在是可行的，通过转基因策略和诊断等位基因检测开辟了分子面包小麦改善的新观点。

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