Spirogyra living in running water is anchored to the substratum by rhizoids that form at the ends of the filaments. A new terminal cell differentiates into a rhizoid cell if the filament is injured. The mode of growth changes from diffuse to tip growth when rhizoid differentiation begins. In this study, we found that a callosic substance was synthesized during rhizoid differentiation. Decreasing the cell turgor, lowering extracellular Ca2 or adding Gd3 , all inhibited the commencement of rhizoid differentiation as well as synthesis of the callose-like substance at the tip of the terminal cell. A callosic substance was also synthesized during formation of the conjugation tube.