Previous work identified the wild barley (Hordeum vulgare ssp. spontaneum) accession CPI-71284-48 as being capable of limiting sodium (Na(+)) accumulation in the shoots under saline hydroponic growth conditions. Quantitative trait locus (QTL) analysis using a cross between CPI-71284-48 and a selection of the cultivated barley (H. vulgare ssp. vulgare) cultivar Barque (Barque-73, a moderate Na(+) excluder) attributed the control of the Na(+) exclusion trait from CPI-71284-48 to a single locus on the short arm of chromosome 7H, which was named HvNax3. The locus reduced shoot Na(+) accumulation by 10-25% in plants grown in 150 mM NaCl. Markers generated using colinearity with rice and Brachypodium, together with the analysis of introgression lines and F(2) and F(3) families, enabled HvNax3 to be mapped to a 1.3-cM interval. Genes from the corresponding rice and Brachypodium intervals encode 16 different classes of proteins and include several plausible candidates for HvNax3. The potential of HvNax3 to provide a useful trait contributing to salinity tolerance in cultivated barley is discussed.