Construction of an Onion (<i>Allium cepa</i> L.) Genetic Linkage Map Using Genotyping-by-Sequencing Analysis with a Reference Gene Set and Identification of QTLs Controlling Anthocyanin Synthesis and Content

Anthocyanins, the pigmented flavonoids responsible for red and blue colors in horticultural products, promote human health by preventing cancers and lowering the risk of cardiovascular disease. Red onions contain several cyanidin- and peonidin-based anthocyanins. In this study, we constructed a single-nucleotide polymorphism (SNP)-based genetic linkage map in an F₂ segregating population derived from a cross between the inbred line &lsquo;SP3B&rsquo; (yellow bulb) and the doubled haploid line &lsquo;H6&prime; (red bulb) to identify quantitative trait loci (QTLs) for total anthocyanin content of onion bulbs using a genotyping-by-sequencing (GBS) analysis based on a reference gene set. A total of 101.9 Gbp of raw sequences were generated using an Illumina HiSeq 2500 system and a total of 1625 SNP loci were identified with the criteria of three minimum depths, lower than 30% missing rate, and more than 5% minor allele frequency. As a result, an onion genetic linkage map consisting of 319 GBS-based SNP loci and 34 high-resolution melting (HRM) markers was constructed with eight linkage groups and a total genetic distance of 881.4 cM. In addition, the linkage groups were assigned to corresponding chromosomes by comparison with the reference genetic map OH1&times;5225 through marker development based on common transcripts. The analysis revealed one major QTL, <i>qAS7.1</i>, for anthocyanin synthesis and two significant QTLs, <i>qAC4.1</i> and <i>qAC4.2</i>, for anthocyanin content. The QTL <i>qAS7.1</i>, located on chromosome 7 with a phenotypic variation of 87.61%, may be a dihydroflavonol 4-reductase (<i>DFR</i>) gene that determines whether the bulb color is red or yellow. The QTLs <i>qAC4.1</i> and <i>qAC4.2</i> are separately positioned on chromosome 4 with <i>R</i>² values of 19.43% and 26.28%, respectively. This map and QTL information will contribute to marker development and breeding for high anthocyanin content in bulb onion.