Cytosolic malate and oxaloacetate activate S-type anion channels in Arabidopsis guard cells.

Abstract

Intracellular malate-starch interconversion plays an important role in stomatal movements. We investigated whether malate or oxaloacetate from the cytosolic membrane side regulate anion channels in the plasma membrane of Arabidopsis thaliana guard cells. Physiological concentrations of cytosolic malate have been reported in the range of 0.4-3mM in leaf cells. Guard cell patch clamp and two-electrode oocyte voltage-clamp experiments were pursued. We show that a concentration of 1mM cytosolic malate greatly activates S-type anion channels in Arabidopsis thaliana guard cells. Interestingly, 1mM cytosolic oxaloacetate also activates S-type anion channels. Malate activation was abrogated at 10mM malate and in SLAC1 anion channel mutant alleles. Interestingly, malate activation of S-type anion currents was disrupted at below resting cytosolic-free calcium concentrations ([Ca2+ ]cyt ), suggesting a key role for basal [Ca2+ ]cyt signaling. Cytosolic malate was not able to directly activate or enhance SLAC1-mediated anion currents in Xenopus oocytes, whereas in positive controls, cytosolic NaHCO3 enhanced SLAC1 activity, suggesting that malate may not directly modulate SLAC1. Cytosolic malate activation of S-type anion currents was impaired in ost1 and in cpk5/6/11/23 quadruple mutant guard cells. Together these findings show that these cytosolic organic anions function in guard cell 'plasma membrane' ion channel regulation.