| Summary: | Plants are affected by the features of their surrounding environment, such as climate change and air pollution caused by anthropogenic activities. In particular, agricultural production is highly sensitive to environmental characteristics. Since no environmental factor is independent, the interactive effects of these factors on plants are essential for agricultural production. In this context, the interactive effects of ozone (O<sub>3</sub>) and supraoptimal temperatures remain unclear. Here, we investigated the physiological and stomatal characteristics of leaf mustard (<i>Brassica juncea</i> L.) in the presence of charcoal-filtered (target concentration, 10 ppb) and elevated (target concentration, 120 ppb) O<sub>3</sub> concentrations and/or optimal (22/20 °C day/night) and supraoptimal temperatures (27/25 °C). Regarding physiological characteristics, the maximum rate of electron transport and triose phosphate use significantly decreased in the presence of elevated O<sub>3</sub> at a supraoptimal temperature (OT conditions) compared with those in the presence of elevated O<sub>3</sub> at an optimal temperature (O conditions). Total chlorophyll content was also significantly affected by supraoptimal temperature and elevated O<sub>3</sub>. The chlorophyll <i>a</i>/<i>b</i> ratio significantly reduced under OT conditions compared to C condition at 7 days after the beginning of exposure (DAE). Regarding stomatal characteristics, there was no significant difference in stomatal pore area between O and OT conditions, but stomatal density under OT conditions was significantly increased compared with that under O conditions. At 14 DAE, the levels of superoxide (O<sub>2</sub><sup>-</sup>), which is a reactive oxygen species, were significantly increased under OT conditions compared with those under O conditions. Furthermore, leaf weight was significantly reduced under OT conditions compared with that under O conditions. Collectively, these results indicate that temperature is a key driver of the O<sub>3</sub> response of <i>B. juncea</i> via changes in leaf physiological and stomatal characteristics.
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