Chloroplast-localized BICAT proteins shape stromal calcium signals and are required for efficient photosynthesis

The photosynthetic machinery of plants must be regulated to maximize the efficiency of light reactions and CO₂ fixation. Changes in free Ca²⁺ in the stroma of chloroplasts have been observed at the transition between light and darkness, and also in response to stress stimuli. Such Ca²⁺ dynamics have been proposed to regulate photosynthetic capacity. However, the molecular mechanisms of Ca²⁺ fluxes in the chloroplasts have been unknown. By employing a Ca²⁺ reporter-based approach, we identified two chloroplast-localized Ca²⁺ transporters in Arabidopsis thaliana, BICAT1 and BICAT2, that determine the amplitude of the darkness-induced Ca²⁺ signal in the chloroplast stroma. BICAT2 mediated Ca²⁺ uptake across the chloroplast envelope, and its knockout mutation strongly dampened the dark-induced [Ca²⁺ ]_stroma signal. Conversely, this Ca²⁺ transient was increased in knockout mutants of BICAT1, which transports Ca²⁺ into the thylakoid lumen. Knockout mutation of BICAT2 caused severe defects in chloroplast morphology, pigmentation and photosynthetic light reactions, rendering bicat2 mutants barely viable under autotrophic growth conditions, while bicat1 mutants were less affected. These results show that BICAT transporters play a role in chloroplast Ca²⁺ homeostasis. They are also involved in the regulation of photosynthesis and plant productivity. Further work will be required to reveal whether the effect on photosynthesis is a direct result of their role as Ca²⁺ transporters.