Electron donation to photosystem I was studied in highly resolved particles from spinach. Divalent cations increased the efficiency of electron donation from spinach plastocyanin to P700(+) through a decrease in the apparent K(m) for plastocyanin. Cytochrome f was not an efficient electron donor for P700(+) in the presence or absence of divalent cations. Cytochrome f photooxidation could be observed in the presence of both plastocyanin and divalent cations.The efficiencies of electron donors from eukaryotic and prokaryotic algae to P700(+) were also examined. Divalent cations enhanced the effectiveness of electron donors from eukaryotic organisms, while inhibiting electron donors from prokaryotic organisms. The prokaryotic electron donors were also much more efficient donors than were the electron donors from eukaryotic organisms. A correlation between the K(m) for the electron donor and its isoelectric point suggests that the net charge on the donor protein is a major determinant of the efficiency for electron donation. The data presented raise interesting questions with respect to the evolution of electron donation to photosystem I and the possibility of an additional electron carrier between plastocyanin and P700(+).