Aqueous solutions of 2-thiouracil (TU) were applied selectively either to the growing point or to the leaves of the long-day plant Hyoscyamus niger in order to determine whether this antimetabolite has an effect on the synthesis of the floral stimulus in the leaves. Applications to the growing point were made by means of a small glass tube covering the shoot apex; application to the leaves was performed by vacuum infiltration. In all experiments all leaves except the three youngest fully expanded leaves and the 8-10 youngest primordia were removed before application. Plants were recorded as having initiated flowers when flower primordia were visible under a dissection microscope 5 weeks after the experiment.TU was inhibitory to photoperiodic induction by long-days of 16 hours when applied to the growing point during the second 8 hours of the daily photoperiod. A concentration of 5·10(-3) M of TU fully suppressed flowering without significant inhibition of leaf primordia increment; however, leaves developing from treated primordia had reduced leaf blades. These results are in agreement with findings already published by other investigators.However, when the leaves were infiltrated by TU, the antimetabolite did not inhibit photoperiodic induction but on the contrary initiated flowering even under short-day conditions. This effect was investigated in more detail by repeated daily infiltrations of TU-solutions in concentrations of 10(-5)-10(-2) M during the second part of an 8 hour photoperiod up to 5 following days. Even after one single infiltration of a 10(-4) M solution 18% of the treated plants were flowering; the percentage of flowering plants increased with increasing concentrations of TU and number of days of application up to approximately 80%. In no case was a flower initiation of 100% obtained. Leaves developing from primordia after infiltration of the leaves with TU have reduced and deformed leaf blades, indicating that TU is transported to the shoot apex to some extent.Some possible explanations of this inductive effect of TU were tested experimentally. Oxygen uptake of the leaves was not decreased and the respiratory quotient was not affected by TU. Photoperiodic induction is not stimulated by low concentrations of TU when applied to the growing point. Infiltration of the leaves by solutions of 2,4-dinitrophenol (10(-4) M) and sodium azide (10(-3) M) had no inductive effect under short-day conditions; a single complete defoliation (except for the 8-10 youngest primordia) is also not inductive. Under short-day conditions additional leaves remaining on the plant that were not infiltrated by TU decreased the percentage of flowering plants but did not fully suppress flower initiation.From these results it is concluded that TU does not act by inhibition of particular metabolic processes concerned in flower initiation or by inhibition of the synthesis of an inhibitor. We suggest that application of TU may lead to synthesis of a floral stimulus in the leaves under short-day conditions also.