Tropospheric ozone is a major air pollutant affecting plants worldwide. Plants in northern regions can display more ozone injury than plants at lower latitudes despite lower ozone levels. Larger ozone influx and shorter nights have been suggested as possible causes. However, the effects of the dim light present during northern summer nights have not been investigated. Young Trifolium subterraneum plants kept in environmentally controlled growth rooms under long day (10 h bright light, 14 h dim light) or short day (10 h bright light, 14 h darkness) conditions were exposed to 6 h of 70 ppb ozone during daytime for three consecutive days. Leaves were visually inspected and imaged in vivo using thermal imaging before and after the daily exposure. In long-day-treated plants, visible foliar injury within 1 week after exposure was more severe. Multivariate statistical analyses showed that the leaves of ozone-exposed long-day-treated plants were also warmer with more homogeneous temperature distributions than exposed short day and control plants, suggesting reduced transpiration. Temperature disruptions were not restricted to areas displaying visible damage and occurred even in leaves with only slight visible injury. Ozone did not affect the leaf temperature of short-day-treated plants. As all factors influencing ozone influx were the same for long- and short-day-treated plants, only the dim nocturnal light could account for the different ozone sensitivities. Thus, the twilight summer nights at high latitudes may have a negative effect on repair and defence processes activated after ozone exposure, thereby enhancing sensitivity.