Electrostatic interactions (EI) are an important factor for phase transitions between lamellar liquid-crystalline (L-alpha) and inverse bicontinuous cubic (Q(II)) phases. We investigated the low pH-induced L-alpha to double-diamond cubic (Q(II)(D)) phase transition in dioleoylphosphatidylserine (DOPS)/monoolein (MO) using time-resolved small-angle X-ray scattering. Using a stopped-flow apparatus, a suspension of liposomes (multilamellar vesicles (MLVs) or large unilamellar vesicles (LUVs)) of 20%-DOPS/80%-MO membrane at neutral pH was rapidly mixed with a low pH buffer, and then the structural change of the membranes in the resultant suspension was observed as a function of time (i.e., pH-jump experiment). At the initial step, the L-alpha phase was directly transformed into the hexagonal II (H-II) phase, and subsequently, the H-II phase slowly converted into the Q(II)(D) phase. We obtained the rate constants of the initial step (i.e., the L-alpha to H-II phase transition) and of the second step (i.e., the H-II to Q(II)(D) phase transition) using the non-negative matrix factorization method. The rate constant of the initial step was independent of the MLV concentration, indicating that single MLVs can convert into the Hu phase without any interaction with other MLVs. On the other hand, the rate constant of the initial step increased with a decrease in pH, 0.041 s(-1) at pH 2.6 and 0.013 s(-1) at pH 2.8, and also exhibited a size dependence; for smaller vesicles such as LUVs and smaller MLVs with diameters of similar to 1 mu m, the rate constant was smaller. They were reasonably explained by the classical nucleation theory. These results provide the first experimental evidence of the total kinetics of EI-induced L-alpha/Q(II) phase transitions.