Nazareno Eric S, Fiedler Jason, Miller Marisa E, Figueroa Melania, Kianian Shahryar F
Department of Plant Pathology, University of Minnesota, St. Paul, MN, USA.
US Department of Agriculture-Agricultural Research Service, Cereal Crops Research Unit, Fargo, ND, USA.
Theor Appl Genet. 2022 Oct;135(10):3307-3321. doi: 10.1007/s00122-022-04128-6. Epub 2022 Aug 27.
We mapped three adult plant resistance (APR) loci on oat chromosomes 4D and 6C and developed flanking KASP/PACE markers for marker-assisted selection and gene pyramiding. Using sequence orthology search and the available oat genomic and transcriptomic data, we surveyed these genomic regions for genes that may control disease resistance. Sources of durable disease resistance are needed to minimize yield losses in cultivated oat caused by crown rust (Puccinia coronata f. sp. avenae). In this study, we developed five oat recombinant inbred line mapping populations to identify sources of adult plant resistance from crosses between five APR donors and Otana, a susceptible variety. The preliminary bulk segregant mapping based on allele frequencies showed two regions in linkage group Mrg21 (Chr4D) that are associated with the APR phenotype in all five populations. Six markers from these regions in Chr4D were converted to high-throughput allele specific PCR assays and were used to genotype all individuals in each population. Simple interval mapping showed two peaks in Chr4D, named QPc.APR-4D.1 and QPc.APR-4D.2, which were detected in the OtanaA/CI4706-2 and OtanaA/CI9416-2 and in the Otana/PI189733, OtanaD/PI260616, and OtanaA/CI8000-4 populations, respectively. These results were validated by mapping two entire populations, Otana/PI189733 and OtanaA/CI9416, genotyped using Illumina HiSeq, in which polymorphisms were called against the OT3098 oat reference genome. Composite interval mapping results confirmed the presence of the two quantitative trait loci (QTL) located on oat chromosome 4D and an additional QTL with a smaller effect located on chromosome 6C. This mapping approach also narrowed down the physical intervals to between 5 and 19 Mb, and indicated that QPc.APR-4D.1, QPc.APR-4D.2, and QPc.APR-6C explained 43.4%, 38.5%, and 21.5% of the phenotypic variation, respectively. In a survey of the gene content of each QTL, several clusters of disease resistance genes that may contribute to APR were found. The allele specific PCR markers developed for these QTL regions would be beneficial for marker-assisted breeding, gene pyramiding, and future cloning of resistance genes from oat.
我们在燕麦4D和6C染色体上定位了三个成株抗性(APR)位点,并开发了侧翼KASP/PACE标记用于标记辅助选择和基因聚合。利用序列同源性搜索以及现有的燕麦基因组和转录组数据,我们在这些基因组区域中查找可能控制抗病性的基因。需要持久抗病性来源,以尽量减少冠锈病(Puccinia coronata f. sp. avenae)导致的栽培燕麦产量损失。在本研究中,我们构建了五个燕麦重组自交系作图群体,以从五个APR供体与感病品种Otana的杂交后代中鉴定成株抗性来源。基于等位基因频率的初步混合分离群体作图显示,连锁群Mrg21(Chr4D)中有两个区域与所有五个群体的APR表型相关。来自Chr4D这些区域的六个标记被转化为高通量等位基因特异性PCR分析方法,并用于对每个群体中的所有个体进行基因分型。简单区间作图显示Chr4D中有两个峰值,分别命名为QPc.APR - 4D.1和QPc.APR - 4D.2,在OtanaA/CI4706 - 2和OtanaA/CI9416 - 2群体中以及在Otana/PI189733、OtanaD/PI260616和OtanaA/CI8000 - 4群体中分别检测到。通过对使用Illumina HiSeq进行基因分型的两个完整群体Otana/PI189733和OtanaA/CI9416进行作图验证了这些结果,其中针对OT3098燕麦参考基因组鉴定了多态性。复合区间作图结果证实了位于燕麦4D染色体上的两个数量性状位点(QTL)以及位于6C染色体上一个效应较小的额外QTL的存在。这种作图方法还将物理区间缩小到5至19 Mb之间,并表明QPc.APR - 4D.1、QPc.APR - 4D.2和QPc.APR - 6C分别解释了43.4%、38.5%和21.5%的表型变异。在对每个QTL的基因内容进行调查时,发现了几个可能有助于APR的抗病基因簇。为这些QTL区域开发的等位基因特异性PCR标记将有利于标记辅助育种、基因聚合以及未来从燕麦中克隆抗性基因。