Department of Cell Biology and Genetics, University of Alcalá, Campus Universitario, Ctra. Madrid-Barcelona km 33,600, Alcalá de Henares, 28871 Madrid, Spain.
Theor Appl Genet. 2013 Jan;126(1):203-18. doi: 10.1007/s00122-012-1974-8. Epub 2012 Sep 5.
Two of the domains most widely shared among R genes are the nucleotide binding site (NBS) and protein kinase (PK) domains. The present study describes and maps a number of new oat resistance gene analogues (RGAs) with two purposes in mind: (1) to identify genetic regions that contain R genes and (2) to determine whether RGAs can be used as molecular markers for qualitative loci and for QTLs affording resistance to Puccinia coronata. Such genes have been mapped in the diploid A. strigosa × A. wiestii (Asw map) and the hexaploid MN841801-1 × Noble-2 (MN map). Genomic and cDNA NBS-RGA probes from oat, barley and wheat were used to produce RFLPs and to obtain markers by motif-directed profiling based on the NBS (NBS profiling) and PK (PK profiling) domains. The efficiency of primers used in NBS/PK profiling to amplify RGA fragments was assessed by sequencing individual marker bands derived from genomic and cDNA fragments. The positions of 184 markers were identified in the Asw map, while those for 99 were identified in the MN map. Large numbers of NBS and PK profiling markers were found in clusters across different linkage groups, with the PK profiling markers more evenly distributed. The location of markers throughout the genetic maps and the composition of marker clusters indicate that NBS- and PK-based markers cover partly complementary regions of oat genomes. Markers of the different classes obtained were found associated with the two resistance loci, PcA and R-284B-2, mapped on Asw, and with five out of eight QTLs for partial resistance in the MN map. 53 RGA-RFLPs and 187 NBS/PK profiling markers were also mapped on the hexaploid map A. byzantina cv. Kanota × A. sativa cv. Ogle. Significant co-localization was seen between the RGA markers in the KO map and other markers closely linked to resistance loci, such as those for P. coronata and barley yellow dwarf virus (Bydv) that were previously mapped in other segregating populations.
两个最广泛共享的 R 基因结构域是核苷酸结合位点（NBS）和蛋白激酶（PK）结构域。本研究描述并绘制了一些新的燕麦抗性基因类似物（RGAs），有两个目的：（1）确定包含 R 基因的遗传区域，（2）确定 RGAs 是否可用于定性基因座和 Puccinia coronata 抗性的 QTL 的分子标记。这些基因已经在二倍体 A. strigosa × A. wiestii（Asw 图谱）和六倍体 MN841801-1 × Noble-2（MN 图谱）中进行了定位。使用来自燕麦、大麦和小麦的基因组和 cDNA NBS-RGA 探针产生 RFLP，并通过基于 NBS（NBS 分析）和 PK（PK 分析）结构域的基序定向分析获得标记。通过对来自基因组和 cDNA 片段的单个标记带进行测序，评估了 NBS/PK 分析中用于扩增 RGA 片段的引物的效率。在 Asw 图谱中鉴定了 184 个标记的位置，而在 MN 图谱中鉴定了 99 个标记的位置。在不同连锁群上发现了大量的 NBS 和 PK 分析标记聚类，PK 分析标记分布更均匀。遗传图谱上标记的位置和标记聚类的组成表明，基于 NBS 和 PK 的标记覆盖了燕麦基因组的部分互补区域。从不同类别获得的标记与在 Asw 图谱上定位的两个抗性基因座 PcA 和 R-284B-2 以及 MN 图谱中 8 个部分抗性 QTL 的 5 个相关联。还在六倍体图谱 A. byzantina cv. Kanota × A. sativa cv. Ogle 上定位了 53 个 RGA-RFLP 和 187 个 NBS/PK 分析标记。在 KO 图谱中的 RGA 标记与其他紧密连锁的抗性基因座标记（如以前在其他分离群体中定位的 P. coronata 和大麦黄矮病毒（Bydv）标记）之间观察到显著的共定位。
