Characterization of calcium fluxes across the envelope of intact spinach chloroplasts

Calcium fluxes across the envelope of intact spinach chloroplasts (Spinacia oleracea L.) in the light and in the dark were investigated using the metallochromic indicator arsenazo III. Light induces Ca(2+) influx into chloroplasts. The action spectrum of light-induced Ca(2+) influx and the inhibitory effect of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) indicate an involement of photosynthetic electron transport in this process. The driving force for light-induced Ca(2+) influx is most likely a change in the membrane potential component of the proton motive force. This was demonstrated by the use of agents modifying the membrane potential (lipophilic cations, ionophores, different KCl concentrations). The activation energy of the observed Ca(2+) influx is about 92 kJ mol(-1). Verapamil and nifedipine, two Ca(2+)-channel blockers, have no inhibitory effect on light-induced Ca(2+) influx, but enhance ferricyanide-dependent oxygen evolution. Inhibition of Ca(2+) influx by ruthenium red reduces the light-dependent decrease in stromal NAD(+) level.