Salt tolerance of plants depends on HKT transporters (High-affinity K+T ransporter), which mediate Na+-specific transport or Na+-K+ co-transport. Gene sequences closely related to rice HKT genes were isolated from hexaploid bread wheat (Triticum aestivum) or barley (Hordeum vulgare) for genomic DNA southern hybridization analysis. HKT gene sequences were mapped on chromosomal arms of wheat and barley using wheat chromosome substitution lines and barley–wheat chromosome addition lines. In addition, HKT gene members in the wild diploid wheat ancestors, T. monococcum (Am genome), T. urartu (Au genome), and Ae. tauschii (Dt genome) were investigated. Variation in copy number for individual HKT gene members was observed between the barley, wheat, and rice genomes, and between the different wheat genomes. HKT2;1/2-like, HKT2;3/4-like, HKT1;1/2-like, HKT1;3-like, HKT1;4-like, and HKT1;5-like genes were mapped to the wheat–barley chromosome groups 7, 7, 2, 6, 2, and 4, respectively. Chromosomal regions containing HKT genes were syntenic between wheat and rice except for the chromosome regions containing the HKT1;5-like gene. Potential roles of HKT genes in Na+ transport in rice, wheat, and barley are discussed. Determination of the chromosome locations of HKT genes provides a framework for future physiological and genetic studies investigating the relationships between HKT genes and salt tolerance in wheat and barley.