Elicitor-induced ethylene biosynthesis in tomato cells: characterization and use as a bioassay for elicitor action

The induction of ethylene biosynthesis by an elicitor partially purified from yeast extract was studied in suspension-cultured tomato (Lycopersicon esculentum Mill.) cells. Unstimulated cells produced little ethylene during exponential growth and even less in stationary phase. Treatment with elicitor stimulated ethylene biosynthesis 10-fold to 20-fold in the exponentially growing cells and more than 100-fold in stationary cells. Activities of both 1-aminocyclopropane-1-carboxylate (ACC) synthase, measured in vitro, and ethylene-forming enzyme (EFE), measured in vivo, increased strongly in response to elicitor treatments. During exponential growth, cells contained large pools of ACC, and the elicitor stimulated ethylene biosynthesis primarily through induction of EFE. In the stationary phase, cells contained almost no ACC, and the elicitor stimulated ethylene biosynthesis primarily through its effect on ACC synthase activity. Cordycepin did not affect the increase in activity of ACC synthase but blocked that of EFE, indicating that the former was posttranscriptionally regulated, the latter transcriptionally regulated. Removal of elicitor by washing or inactivation of a biotinylated derivative of the elicitor by complexation with avidin caused a rapid cessation of the increase in ACC synthase activity, suggesting that continuous presence of stimulus is necessary for the response. Using induction of ethylene production to measure amounts of elicitor, it was found that the elicitor disappeared from the incubation medium during the course of the treatment.