Circadian gating of dark-induced increases in chloroplast- and cytosolic-free calcium in Arabidopsis

Abstract

Changes in the spatiotemporal concentration of free Ca²⁺ ([Ca²⁺ ]) in different organelles of the cell contribute to responses of plants to physiological and environmental stimuli. One example are [Ca²⁺ ] increases in the stroma of chloroplasts during light-to-dark transitions; however, the function and mechanisms responsible are unknown, in part because there is a disagreement in the literature concerning whether corresponding dark-induced changes in cytosolic [Ca²⁺ ] ([Ca²⁺ ]_cyt ) can be detected. We have measured changes in [Ca²⁺ ]_cyt upon darkness in addition to the already known dark-induced increases in [Ca²⁺ ]_stroma in the aerial part of the Arabidopsis thaliana plant. These [Ca²⁺ ]_cyt transients depend on the photoperiod and time of day, peaking at anticipated dusk, and are superimposed on daily 24 h oscillations in [Ca²⁺ ]_cyt . We also find that the magnitude of the dark-induced increases in Ca²⁺ in both the cytosol and chloroplasts are gated by the nuclear circadian oscillator. The modulation of the magnitude of dark-induced increases in [Ca²⁺ ]_stroma and [Ca²⁺ ]_cyt by transcriptional regulators in the nucleus that are part of the circadian oscillator demonstrates a new role for the circadian system in subcellular Ca²⁺ signalling, in addition to its role in driving circadian oscillations of [Ca²⁺ ] in the cytosol and chloroplasts.

Keywords: Arabidopsis; calcium signalling; chloroplast; circadian clock; cytosol; light-dark transition.