The leaves of Mimosa pudica L. are well known for their rapid movement when touched. Recently, we were able to isolate an excitatory substance in small quantities from this plant, which consists of three different components (potassium L-malate, magnesium trans-aconitate, and dimethylammonium salt). Many plants close their leaves in the evening, as if to sleep, and open them early in the morning (nyctinastic leaf movement). This circadian rhythm is known to be controlled by the biological clock of such plants. Extensive studies on other nyctinastic plants led to the isolation of a variety of leaf-opening substances (LOSs) and leaf-closing substances (LCSs). Based on our experiments on these bioactive substances, we found that the circadian rhythmic leaf movement of these plants is initiated by the regulated balance of LOSs and LCSs. The balance of concentration between the two leaf-movement factors (LMFs) is inversed during the day. The glycoside-type LMF is hydrolyzed with beta-glucosidase, the activity of which is regulated by the biological clock. The circadian rhythm observed in the leaf movement is introduced by activation of beta-glucosidase regulated by the biological clock.