The role of potassium (K(+)) transporters in high- and low-affinity K(+) uptake was examined in roots of intact barley (Hordeum vulgare) and Arabidopsis (Arabidopsis thaliana) plants by use of (42)K radiotracing, electrophysiology, pharmacology, and mutant analysis. Comparisons were made between results from barley and five genotypes of Arabidopsis, including single and double knockout mutants for the high-affinity transporter, AtHAK5, and the Shaker-type channel, AtAKT1. In Arabidopsis, steady-state K(+) influx at low external K(+) concentration ([K(+)]ext = 22.5 µm) was predominantly mediated by AtAKT1 when high-affinity transport was inhibited by ammonium, whereas in barley, by contrast, K(+) channels could not operate below 100 µm. Withdrawal of ammonium resulted in an immediate and dramatic stimulation of K(+) influx in barley, indicating a shift from active to passive K(+) uptake at low [K(+)]ext and yielding fluxes as high as 36 µmol g (root fresh weight)(-1) h(-1) at 5 mm [K(+)]ext, among the highest transporter-mediated K(+) fluxes hitherto reported. This ammonium-withdrawal effect was also established in all Arabidopsis lines (the wild types, atakt1, athak5, and athak5 atakt1) at low [K(+)]ext, revealing the concerted involvement of several transport systems. The ammonium-withdrawal effect coincided with a suppression of K(+) efflux and a significant hyperpolarization of the plasma membrane in all genotypes except athak5 atakt1, could be sustained over 24 h, and resulted in increased tissue K(+) accumulation. We discuss key differences and similarities in K(+) acquisition between two important model systems and reveal novel aspects of K(+) transport in planta.