Biosystematics Group, Wageningen University and Research Centre, Generaal Foulkesweg 37, Wageningen, The Netherlands.
Theor Appl Genet. 2010 Feb;120(4):785-96. doi: 10.1007/s00122-009-1199-7. Epub 2009 Nov 10.
Mapping resistance genes is usually accomplished by phenotyping a segregating population for the resistance trait and genotyping it using a large number of markers. Most resistance genes are of the NBS-LRR type, of which an increasing number is sequenced. These genes and their analogs (RGAs) are often organized in clusters. Clusters tend to be rather homogenous, viz. containing genes that show high sequence similarity with each other. From many of these clusters the map position is known. In this study we present and test a novel method to quickly identify to which cluster a new resistance gene belongs and to produce markers that can be used for introgression breeding. We used NBS profiling to identify markers in bulked DNA samples prepared from resistant and susceptible genotypes of small segregating populations. Markers co-segregating with resistance can be tested on individual plants and directly used for breeding. To identify the resistance gene cluster a gene belongs to, the fragments were sequenced and the sequences analyzed using bioinformatics tools. Putative map positions arising from this analysis were validated using markers mapped in the segregating population. The versatility of the approach is demonstrated with a number of populations derived from wild Solanum species segregating for P. infestans resistance. Newly identified P. infestans resistance genes originating from S. verrucosum, S. schenckii, and S. capsicibaccatum could be mapped to potato chromosomes 6, 4, and 11, respectively.
抗性基因的定位通常通过对分离群体进行抗性表型分析,并使用大量标记对其进行基因型分析来完成。大多数抗性基因属于 NBS-LRR 类型,其中越来越多的基因被测序。这些基因及其类似物(RGAs)通常组织在簇中。簇往往是相当同质的,即包含彼此之间具有高度序列相似性的基因。这些簇中的许多簇的图谱位置已知。在这项研究中,我们提出并测试了一种新的方法,可以快速确定新的抗性基因所属的簇,并产生可用于基因渐渗育种的标记。我们使用 NBS 分析来鉴定从小的分离群体的抗性和敏感基因型制备的混池 DNA 样本中的标记。与抗性共分离的标记可以在个体植株上进行测试,并直接用于育种。为了确定抗性基因簇所属的基因,我们对片段进行测序,并使用生物信息学工具分析序列。从该分析中得出的假定图谱位置使用在分离群体中映射的标记进行验证。该方法的多功能性通过来自野生茄属物种的多个分离群体进行验证,这些群体对晚疫病菌具有抗性。从 S. verrucosum、S. schenckii 和 S. capsicibaccatum 中鉴定出的新的晚疫病菌抗性基因可以分别映射到马铃薯染色体 6、4 和 11 上。