Knowledge of the nature and extent of karyotypic differences between species provides insight into the evolutionary history of the genomes in question and, in the case of closely related species, the potential for genetic exchange between taxa. We constructed high-density genetic maps of the silverleaf sunflower (Helianthus argophyllus) and Algodones Dune sunflower (H. niveus ssp. tephrodes) genomes and compared them to a consensus map of cultivated sunflower (H. annuus) to identify chromosomal rearrangements between species. The genetic maps of H. argophyllus and H. niveus ssp. tephrodes included 17 linkage groups each and spanned 1337 and 1478 cM, respectively. Comparative analyses revealed greater divergence between H. annuus and H. niveus ssp. tephrodes (13 inverted segments, 18 translocated segments) than between H. annuus and H. argophyllus (10 inverted segments, 8 translocated segments), consistent with their known phylogenetic relationships. Marker order was conserved across much of the genome, with 83 and 64% of the H. argophyllus and H. niveus ssp. tephrodes genomes, respectively, being syntenic with H. annuus. Population genomic analyses between H. annuus and H. argophyllus, which are sympatric across a portion of the natural range of H. annuus, revealed significantly elevated genetic structure in rearranged portions of the genome, indicating that such rearrangements are associated with restricted gene flow between these two species.C HROMOSOMAL rearrangements are of considerable interest because they are often associated with barriers to gene flow between related species, either due to their direct effects on the fitness of heterozygotes or through the indirect effects of genic barriers embedded within them (White 1978;Barton and Bengtsson 1986;Rieseberg et al. 1995bRieseberg et al. , 1999Rieseberg 2001;Navarro and Barton 2003;Kirkpatrick and Barton 2006; reviewed in Faria and Navarro 2010;Gimenez et al. 2012). As such, detailed information on karyotypic differences between species provides insight into the nature of reproductive isolation and, more pragmatically, informs attempts to introgress beneficial alleles from wild species into crop gene pools (Chetelat and Meglic 2000;Foulongne et al. 2003;Dirlewanger et al. 2004). An improved understanding of synteny across species can also facilitate the identification and localization of functionally important genes in a taxon of interest through the extrapolation of gene order from model species (e.g., Choi et al. 2004;Dilbirligi et al. 2006).The genus Helianthus, which is composed of 49 species native to the Americas (Timme et al. 2007) and includes cultivated sunflower (Helianthus annuus L.; 2n = 2x = 34; hereafter referred to as ANN), has emerged as a model for genetic studies of adaptation, hybridization, and speciation (Rieseberg et al. 1995a,b;Lai et al. 2005;Reagon and Snow 2006;Massinga et al. 2009;Gutierrez et al. 2010;Vekemans 2010;Roumet et al. 2013). Insight into the nature and extent of reproductive barriers within Helianthus will ...