Many studies on salinity stress assume that responses in hydroponics mimic those in soil. However, interactions between the soil solution and the soil matrix can affect responses to salinity stress. This study compared responses to salinity in hydroponics and soil, using two varieties of barley (Hordeum vulgare L.). The responses to salinity caused by high concentrations of Na+ and Cl– were compared to assess any consistent differences between hydroponics and soil associated with a cation and an anion that contribute to salinity stress. Concentrated nutrient solutions were also used to assess the effects of osmotic stress. The effects of salinity differed between the hydroponic and soil systems. Differences between barley cultivars in growth, tissue moisture content and ionic composition were not apparent in hydroponics, whereas significant differences occurred in soil. Growth reductions were greater under hydroponics than in soil at similar electrical conductivity values, and the uptake of Na+ and Cl– was also greater. The relative importance of ion exclusion and osmotic stress varied. In soil, ion exclusion tended to be more important at low to moderate levels of stress (EC at field capacity up to 10 dS m–1) but osmotic stress became more important at higher stress levels. High external concentrations of Cl– had similar adverse effects as high concentrations of Na+, suggesting that Cl– toxicity may reduce growth. Fundamental differences in salinity responses appeared between soil and solution culture, and the importance of the different mechanisms of damage varies according to the severity and duration of the salt stress.