The effects of the polyamines spermidine and 1,3-diaminopropane on ethylene biosynthesis and chlorophyll (Chl) loss were studied in peeled leaves of oat (Avena sativa L., var. Victory) incubated in the dark. Peeling off the epidermal cells induces an increase in 1-aminocyclopropane-lcarboxylate (ACC) synthase activity, resulting in an enhanced ACC and ethylene formation. Both polyamines inhibit ethylene biosynthesis from methioine by inhibiting ACC synthase activity and, more effectively, the conversion of ACC to ethylene. They also inhibit Chi loss occurring between 24 and 48 h of dark incubation; but, as shown by inhibitor experiments, inhibition of Chl loss does not result from inhibition of ethylene formation. Ethylene production and Chl loss, both associated with senescence, require membrane integrity, thus, treatments which promote deterioration of membranes inhibit both processes. Ca" in the incubation medium competitively reduces the polyamine-mediated inhibition of ACC conversion and Chl loss. The data suggest that polyamines initially attach to membranes, thereby inducing changes which, in turn, lead to inhibition of ethylene biosynthesis and retardation of senescence.In the preceding study, we investigated the effects of exogenous Dap4 and Spd on Chl loss, protease activity, and ethylene production in senescing oat leaves, in relation to the endogenous levels of PAs (26). We found an accumulation of Dap after application of exogenous Spd, presumably due to the activity of PA oxidase. We described the inhibitory effect of Dap on protease activitJr, Chl breakdown, and ethylene production. The observed Ca + antagonism of these senescence processes suggested that Dap exerts its influence through an initial interaction with specific negatively charged sites on membranes. We have here extended our study to investigate the possible mode of action of Dap and Spd, including effects on the biosynthesis of ethylene. Formation of ethylene is generally associated with leaf senescence in the dark, although an obligatory connection between the two seems not to exist (1 1).