Stomatal pores on the leaf surface are the gateways for gas exchange between plants and the atmosphere, which is regulated mainly by the S-type anion channel SLAC1. However, the gene encoding the main S-type anion channel SLAC1 in pear and its genetic characteristics remain unknown. In this study, Pbr015894.1 was identified as the candidate for PbrSLAC1 in pear, and it was found to be expressed abundantly in leaves, particularly in the guard cells. Virus-induced gene silencing experiments indicated that stomatal closure was achieved by a change in cell turgor instigated by PbrSLAC1 channel transport of NO3- in pear leaves and induced by abscisic acid. Furthermore, the expression of PbrSLAC1 in Arabidopsis slac1-3 and slac1-4 rescued the defective NO3- transport seen in these mutants, pointing to its role in anion transport. Fluorescence microscopy suggested that PbrSLAC1 was localized in the plasma membrane, and a dual-luciferase assay system demonstrated an interaction between PbrSLAC1 and PbrSnRK2.3/2.8. Moreover, anion conductance mediated by PbrSLAC1 was activated by PbrSnRK2.3 in Xenopus laevis oocytes and the channel showed greater permeability for nitrate than chloride, sulfate, or malate ions. Taken together, these results demonstrate that PbrSLAC1, an anion channel regulated by PbrSnRK2.3, is involved in stomatal closure by mediating the efflux of NO3- in pear leaf.
Keywords: Anion channel; Guard cell; Nitrate; PbrSLAC1; PbrSnRK2.3.
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