SNP-markers in Allium species to facilitate introgression breeding in onion

Abstract

Background: 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.

Results: 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.

Conclusions: 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.

Keywords: A. fistulosum; A. roylei; Allium cepa; Botrytis squamosa; Interspecific hybrids; Transcriptome sequencing.

Fig. 1

Classes of infection by Botrytis squamosa observed on genotypes of the CCxRF population, from left to right: Class 1 one or a few spots on a single leaf; 2 several small spots on one or two leaves; 3 large spots on one or more leaves and 4 more than 50 % of the leaves with spots

Fig. 2

The QTL for resistance to Botrytis squamosa in the the CCxRF population originating from A. roylei identified on chromosome 6 in three independent disease tests. Lines with dashed ends show the LOD region and solid bars represent 1 LOD interval from the maximum LOD scores. Map distances are in cM