Remobilization of inorganic phosphate (P(i)) within a plant is critical for sustaining growth and seed production under external P(i) fluctuation. The barley (Hordeum vulgare) transporter HvPHT1;6 has been implicated in P(i) remobilization. In this report, we expressed HvPHT1;6 in Xenopus laevis oocytes, allowing detailed characterization of voltage-dependent fluxes and currents induced by HvPHT1;6. HvPHT1;6 increased efflux of P(i) near oocyte resting membrane potentials, dependent on external P(i) concentration. Time-dependent inward currents were observed when membrane potentials were more negative than -160 mV, which was consistent with nH(+):HPO(4)(2-) (n > 2) cotransport, based on simultaneous radiotracer and oocyte voltage clamping, dependent upon P(i) concentration gradient and pH. Time- and voltage-dependent inward currents through HvPHT1;6 were also observed for SO(4)(2-)and to a lesser degree for NO(3)(-)Cl(-)but not for malate. Inward and outward currents showed linear dependence on the concentration of external HPO(4)(2-)similar to low-affinity P(i) transport in plant studies. The electrophysiological properties of HvPHT1;6, which locates to the plasma membrane when expressed in onion (Allium cepa) epidermal cells, are consistent with its suggested role in the remobilization of P(i) in barley plants.