A unique variant strain of Chara corallina, which contains little inorganic phosphate in the vacuole ([Pi]v) was isolated. The level of cytoplasmic inorganic phosphate ([Pi]c) in these cells was the same as that in normal cells. Using these unique cells, we studied the change in [Pi]c and the effect of Pi on the activities of electrogenic H(+)-pumps associated with the plasma membrane and tonoplast. Upon illumination, the plasma membrane of C. corallina became hyperpolarized by 15 mV, the pH of the vacuolar sap decreased by 0.5 unit, and [Pi]c decreased by 30% with a similar time course. The activities of the electrogenic H (+)-pump in the plasma membrane and the ATP and PPi-dependent H(+)-transport in the tonoplast were noncompetitively inhibited by Pi with Ki values of, in the order given, 21.3 mM, 22.1 mM and 37.7 mM. From the kinetics study we calculated that the electrogenic H(+)-pump in the plasma membrane and the ATP and PPi-dependent H(+) transport in the tonoplast were activated by, again in this order, 13%, 13% and 9%, in accordance with the decrease in [Pi]c. We propose that the change in [Pi]c is one of the regulators of photosynthesis-mediated activation of the H(+)-pumps in the plasma membrane and the tonoplast in C. corallina upon illumination.