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A chimeric CaHAK1–LeHAK5 transporter with only 15 amino acids of CaHAK1 in the N-terminus mediates high-affinity K⁺ uptake in yeast cells. Kinetic and expression analyses strongly suggest that LeHAK5 mediates a significant proportion of the high-affinity K⁺ uptake shown by K⁺-starved tomato (Solanum lycopersicum) plants. The development of high-affinity K⁺ uptake, putatively mediated by LeHAK5, was correlated with increased LeHAK5 mRNA levels and a more negative electrical potential difference across the plasma membrane of root epidermal and cortical cells. However, this increase in high-affinity K⁺ uptake was not correlated with the root K⁺ content. Thus, (i) growth conditions that result in a hyperpolarized root plasma membrane potential, such as K⁺ starvation or growth in the presence of NH₄ ⁺, but which do not decrease the K⁺ content, lead to increased LeHAK5 expression …