Malate-Regulated Channels Permeable to Anions in Vacuoles of Arabidopsis thaliana

Abstract

Anion channels in isolated vacuoles of Arabidopsis thaliana cultured cells were studied by means of the patch clamp technique in the whole-vacuole configuration. In symmetrical 100 mM KCl, a high resistance of the membrane at positive potentials inside the vacuole was observed. In symmetrical 100 mM K₂-malate positive potentials inside the vacuole elicited slowly developing inward currents, due to the opening of channels, which, according to measurements of reversal potential, are selective for malate. The activation potential of the channels shifted as a function of the cytoplasmic malate concentration, but it was always such that the channels opened only to mediate malate influx into the vacuole. The channels were also permeable to succinate, fumarate and, to a lesser extent, oxaloacetate. In vacuoles preincubated with cytoplas- mic malate, inward currents were also elicited in the presence of KCl or KNO₃ at the cytoplasmic side of the tonoplast. Malate channels were different from the cation slow vacuolar-type channels with regard to their sensitivity to changes in the cytoplasmic concentrations of Ca²⁺ and ATP, and in temperature between 10 and 20ºC.