Cell wall-free protoplasts of P. cyclopium could regenerate a cell wall and form mycelia in liquid culture with high rates of viability. When calcium was added to the medium, protoplasts displayed biphasic accumulation with an immediate metabolism-independent adsorption phase, followed by slow metabolism-dependent uptake. Exposure of the protoplasts to Ca2+ for periods of 2 min, followed by incubation in calcium-free medium for 24 hours, was sufficient to induce conidiation with morphogenetic events parallel to those found in cultures containing calcium throughout the incubation period, and similar to those reported in cultures inoculated from conidia. The conidiation event caused by short exposure to calcium could be reversed, within 2 hours of Ca2+ addition, by a brief treatment with the specific calcium chelating agent BAPTA (100 microM), which removed 65 to 75% of the total cell calcium. The results implicate the membrane-bound calcium fraction in the process of conidiation induction.