Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China.
Theor Appl Genet. 2011 Mar;122(4):795-803. doi: 10.1007/s00122-010-1487-2. Epub 2010 Nov 21.
Scab, caused by Cladosporium cucumerinum, is an important disease of cucumber, Cucumis sativus. In this study, we conducted fine genetic mapping of the single dominant scab resistance gene, Ccu, with 148 F(9) recombinant inbred lines (RILs) and 1,944 F(2) plants derived from the resistant cucumber inbred line 9110Gt and the susceptible line 9930, whose draft genome sequence is now available. A framework linkage map was first constructed with simple sequence repeat markers placing Ccu into the terminal 670 kb region of cucumber Chromosome 2. The 9110Gt genome was sequenced at 5× genome coverage with the Solexa next-generation sequencing technology. Sequence analysis of the assembled 9110Gt contigs and the Ccu region of the 9930 genome identified three insertion/deletion (Indel) markers, Indel01, Indel02, and Indel03 that were closely linked with the Ccu locus. On the high-resolution map developed with the F(2) population, the two closest flanking markers, Indel01 and Indel02, were 0.14 and 0.15 cM away from the target gene Ccu, respectively, and the physical distance between the two markers was approximately 140 kb. Detailed annotation of the 180 kb region harboring the Ccu locus identified a cluster of six resistance gene analogs (RGAs) that belong to the nucleotide binding site (NBS) type R genes. Four RGAs were in the region delimited by markers Indel01 and Indel02, and thus were possible candidates of Ccu. Comparative DNA analysis of this cucumber Ccu gene region with a melon (C. melo) bacterial artificial chromosome (BAC) clone revealed a high degree of micro-synteny and conservation of the RGA tandem repeats in this region.
疮痂病由半裸球腔菌(Cladosporium cucumerinum)引起,是黄瓜(Cucumis sativus)的重要病害。本研究利用来自抗疮痂病黄瓜自交系 9110Gt 和感病自交系 9930 的 148 个 F9 重组自交系(RIL)和 1944 个 F2 代植株,对单一显性抗疮痂病基因 Ccu 进行了精细遗传作图。首先利用简单重复序列标记构建了一个框架连锁图谱,将 Ccu 定位在黄瓜第 2 号染色体的末端 670kb 区域。9110Gt 基因组采用 Solexa 新一代测序技术以 5×基因组覆盖度进行测序。对组装的 9110Gt 连续序列和 9930 基因组的 Ccu 区域进行序列分析,鉴定出 3 个插入/缺失(Indel)标记 Indel01、Indel02 和 Indel03,它们与 Ccu 位点紧密连锁。在利用 F2 群体构建的高分辨率图谱上,两个最接近的侧翼标记 Indel01 和 Indel02 与目标基因 Ccu 的距离分别为 0.14 和 0.15cM,两个标记之间的物理距离约为 140kb。对包含 Ccu 基因座的 180kb 区域进行详细注释,鉴定出一个包含 6 个抗性基因类似物(RGA)的簇,这些 RGA 属于核苷酸结合位点(NBS)型 R 基因。4 个 RGA 位于标记 Indel01 和 Indel02 界定的区域内,因此可能是 Ccu 的候选基因。对该黄瓜 Ccu 基因区域与甜瓜(Cucumis melo)细菌人工染色体(BAC)克隆的比较 DNA 分析显示,该区域的 NBS 型 RGA 串联重复具有高度的微同线性和保守性。