The effects of ozone on bean plants pretreated with the SAR activator benzothiadiazole (BTH) have been investigated after fumigations with an acute dose of the pollutant (200 nL x L(-1) for 4 h), carried out at different times from BTH application. BTH pretreatment induced opposite effects on bean susceptibility to O(3), depending on the time elapsed before fumigation. When this time was only 1-2 days, bean plants were more susceptible to O(3) than untreated controls, showing rapid and extensive cell death in both palisade and spongy mesophyll. These damages appeared to be closely correlated with the amount and localization of H(2)O(2) in the leaf tissues. In BTH-pretreated, but not fumigated, plants, H(2)O(2) accumulation occurred in the cell walls and no dead cells were detected, whereas O(3) fumigation of untreated plants produced H(2)O(2) accumulation also inside some palisade mesophyll cells, causing their death. When BTH pretreatments were carried out 5-7 days before fumigation, plants appeared to be more tolerant to O(3) compared to untreated controls. Under these conditions, no visible symptoms of phytotoxicity were observed for at least 2 weeks after fumigation and no H(2)O(2) accumulation was detected. Biochemical assays showed a significant increase in the ascorbate (AA) level, taking place from the fifth to the seventh day after BTH treatment and unaffected by O(3) when given at these times. Ascorbate peroxidase (APX) activity appeared to decrease during the first 2 days after BTH treatment, and the decrease was somewhat enhanced by fumigation. On the contrary, guaiacol peroxidase (GuPX) activity was found to steadily increase up to the fifth day after BTH treatment but showed a bimodal trend upon fumigation. These results suggest that, during the first 1-2 days after BTH application, the H(2)O(2) level is enhanced by O(3) over a critical threshold for cell viability. However, in the absence of the pollutant, H(2)O(2) decreases in the following days under the effect of AA accumulation and increased GuPX activity. As GuPX is known to promote cell wall lignification and protein cross-linking, these effects would protect plasmalemma from O(3) irreversible damage, provided the priming by BTH has been fully developed.