Brotman Y., Silberstein L., Kovalski I., Perin C., Dogimont C., Pitrat M., Klingler J., Thompson A., Perl-Treves R.
Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel.
Theor Appl Genet. 2002 May;104(6-7):1055-1063. doi: 10.1007/s00122-001-0808-x. Epub 2002 Feb 8.
Genomic and cDNA fragments with homology to known disease resistance genes (RGH fragments) were cloned from Cucumis melo using degenerate-primer PCR. Fifteen homologues of the NBS-LRR gene family have been isolated. The NBS-LRR homologues show high divergence and, based on the partial NBS-fragment sequences, appear to include members of the two major subfamilies that have been described in dicot plants, one that possesses a TIR-protein element and one that lacks such a domain. Genomic organization of these sequences was explored by DNA gel-blot analysis, and conservation among other Cucurbitaceae was assessed. Two mapping populations that segregate for several disease and pest resistance loci were used to map the RGH probes onto the melon genetic map. Several NBS-LRR related sequences mapped to the vicinity of genetic loci that control resistance to papaya ringspot virus, Fusarium oxysporum race 1, F. oxysporum race 2 and to the insect pest Aphis gossypii. The utility of such markers for breeding resistant melon cultivars and for cloning the respective R-genes is discussed.
利用简并引物PCR从甜瓜中克隆了与已知抗病基因具有同源性的基因组和cDNA片段(RGH片段)。已分离出15个NBS-LRR基因家族的同源物。这些NBS-LRR同源物表现出高度的差异,基于部分NBS片段序列,似乎包括双子叶植物中已描述的两个主要亚家族的成员,一个具有TIR蛋白元件,另一个缺乏该结构域。通过DNA凝胶印迹分析探索了这些序列的基因组组织,并评估了其他葫芦科植物之间的保守性。使用两个分离多个抗病和抗虫位点的作图群体将RGH探针定位到甜瓜遗传图谱上。几个与NBS-LRR相关的序列定位到控制对木瓜环斑病毒、尖孢镰刀菌1号小种、尖孢镰刀菌2号小种以及棉蚜抗性的遗传位点附近。讨论了此类标记在培育抗病甜瓜品种和克隆相应R基因方面的实用性。