Salinity stress includes both osmotic and ionic toxicity. Sodium homeostasis is influenced by Na(+) uptake and extrusion, vacuolar Na(+) compartmentation and root to shoot Na(+) translocation via transpiration. The knockout mutant of the Arabidopsis heterotrimeric G-protein Gβ subunit, agb1, is hypersensitive to salt, exhibiting a leaf bleaching phenotype. We show that AGB1 is mainly involved in the ionic toxicity component of salinity stress and plays roles in multiple processes of Na(+) homeostasis. agb1 mutants accumulate more Na(+) and less K(+) in both shoots and roots of hydroponically grown plants, as measured by inductively coupled plasma atomic emission spectrometry. agb1 plants have higher root to shoot translocation rates of radiolabelled (24) Na(+) under transpiring conditions, as a result of larger stomatal apertures and increased stomatal conductance. (24) Na(+) tracer experiments also show that (24) Na(+) uptake rates by excised roots of agb1 and wild type are initially equal, but that agb1 has higher net Na(+) uptake at 90 min, implicating possible involvement of AGB1 in the regulation of Na(+) efflux. Calcium alleviates the salt hypersensitivity of agb1 by reducing Na(+) accumulation to below the toxicity threshold. Our results provide new insights into the regulatory pathways underlying plant responses to salinity stress, an important agricultural problem.