SNP discovery and development of a high-density genotyping array for sunflower

Abstract

Recent advances in next-generation DNA sequencing technologies have made possible the development of high-throughput SNP genotyping platforms that allow for the simultaneous interrogation of thousands of single-nucleotide polymorphisms (SNPs). Such resources have the potential to facilitate the rapid development of high-density genetic maps, and to enable genome-wide association studies as well as molecular breeding approaches in a variety of taxa. Herein, we describe the development of a SNP genotyping resource for use in sunflower (Helianthus annuus L.). This work involved the development of a reference transcriptome assembly for sunflower, the discovery of thousands of high quality SNPs based on the generation and analysis of ca. 6 Gb of transcriptome re-sequencing data derived from multiple genotypes, the selection of 10,640 SNPs for inclusion in the genotyping array, and the use of the resulting array to screen a diverse panel of sunflower accessions as well as related wild species. The results of this work revealed a high frequency of polymorphic SNPs and relatively high level of cross-species transferability. Indeed, greater than 95% of successful SNP assays revealed polymorphism, and more than 90% of these assays could be successfully transferred to related wild species. Analysis of the polymorphism data revealed patterns of genetic differentiation that were largely congruent with the evolutionary history of sunflower, though the large number of markers allowed for finer resolution than has previously been possible.

Figure 1. STRUCTURE results plot.

Results of STRUCTURE analysis of the 32 H. annuus individuals based on all SNPs with MAF ≥0.10. A) Depicts the results for K = 2. B) Depicts the results for K = 3. Black bars represent dividers between the six groups: OPV/Landraces, HA-oil, HA-nonoil (HA-NO), RHA-oil, RHA-nonoil (RHA-NO), and wild H. annuus.

Figure 2. Principal coordinates analysis plot.

Plot of the first two principal coordinates for the 32 H. annuus individuals based on all SNPs with MAF ≥0.10. Each data point represents an accession with one of six groups: OPV/Landraces (OPV/LR), HA-oil, HA-nonoil (HA-NO), RHA-oil, RHA-nonoil (RHA-NO), and wild H. annuus.