Scholten Olga E, van Kaauwen Martijn P W, Shahin Arwa, Hendrickx Patrick M, Keizer L C Paul, Burger Karin, van Heusden Adriaan W, van der Linden C Gerard, Vosman Ben
Wageningen UR Plant Breeding, Wageningen University and Research Centre, PO Box 386, Wageningen, 6700 AJ, The Netherlands.
BMC Plant Biol. 2016 Aug 31;16(1):187. doi: 10.1186/s12870-016-0879-0.
Within onion, Allium cepa L., the availability of disease resistance is limited. The identification of sources of resistance in related species, such as Allium roylei and Allium fistulosum, was a first step towards the improvement of onion cultivars by breeding. SNP markers linked to resistance and polymorphic between these related species and onion cultivars are a valuable tool to efficiently introgress disease resistance genes. In this paper we describe the identification and validation of SNP markers valuable for onion breeding.
Transcriptome sequencing resulted in 192 million RNA seq reads from the interspecific F1 hybrid between A. roylei and A. fistulosum (RF) and nine onion cultivars. After assembly, reliable SNPs were discovered in about 36 % of the contigs. For genotyping of the interspecific three-way cross population, derived from a cross between an onion cultivar and the RF (CCxRF), 1100 SNPs that are polymorphic in RF and monomorphic in the onion cultivars (RF SNPs) were selected for the development of KASP assays. A molecular linkage map based on 667 RF-SNP markers was constructed for CCxRF. In addition, KASP assays were developed for 1600 onion-SNPs (SNPs polymorphic among onion cultivars). A second linkage map was constructed for an F2 of onion x A. roylei (F2(CxR)) that consisted of 182 onion-SNPs and 119 RF-SNPs, and 76 previously mapped markers. Markers co-segregating in both the F2(CxR) and the CCxRF population were used to assign the linkage groups of RF to onion chromosomes. To validate usefulness of these SNP markers, QTL mapping was applied in the CCxRF population that segregates for resistance to Botrytis squamosa and resulted in a QTL for resistance on chromosome 6 of A. roylei.
Our research has more than doubled the publicly available marker sequences of expressed onion genes and two onion-related species. It resulted in a detailed genetic map for the interspecific CCxRF population. This is the first paper that reports the detection of a QTL for resistance to B. squamosa in A. roylei.
在洋葱(Allium cepa L.)中,抗病性资源有限。鉴定相关物种如葱(Allium roylei)和大葱(Allium fistulosum)中的抗性来源是通过育种改良洋葱品种的第一步。与抗性相关且在这些相关物种和洋葱品种之间具有多态性的单核苷酸多态性(SNP)标记是有效导入抗病基因的宝贵工具。在本文中,我们描述了对洋葱育种有价值的SNP标记的鉴定和验证。
转录组测序产生了来自葱和大葱种间F1杂种(RF)以及九个洋葱品种的1.92亿条RNA序列读数。组装后,在约36%的重叠群中发现了可靠的SNP。为了对种间三交群体(源自洋葱品种与RF的杂交(CCxRF))进行基因分型,选择了1100个在RF中具有多态性而在洋葱品种中为单态性的SNP(RF-SNP)用于竞争性等位基因特异性PCR(KASP)分析的开发。基于667个RF-SNP标记构建了CCxRF的分子连锁图谱。此外,还针对1600个洋葱-SNP(在洋葱品种间具有多态性的SNP)开发了KASP分析。为洋葱×葱的F2(F2(CxR))构建了第二个连锁图谱,该图谱由182个洋葱-SNP、119个RF-SNP和76个先前定位的标记组成。在F2(CxR)和CCxRF群体中共同分离的标记用于将RF的连锁群分配到洋葱染色体上。为了验证这些SNP标记的实用性,在对洋葱核盘菌抗性进行分离的CCxRF群体中进行了数量性状基因座(QTL)定位,结果在葱的6号染色体上发现了一个抗性QTL。
我们的研究使公开可用的洋葱表达基因和两个与洋葱相关物种的标记序列增加了一倍多。它为种间CCxRF群体生成了详细的遗传图谱。这是第一篇报道在葱中检测到对核盘菌抗性QTL的论文。