AbstractChlorophyll fluorescence induction was used as a probe to detect herbicide detoxification in tolerant or susceptible wheat cultivars. Experimental conditions have been carefully examined for establishing detoxification kinetics of chlorotoluron and metribuzin, two photosystem‐II‐inhibiting herbicides. After a root treatment, leaves were cut, placed in glass tubes and maintained in the dark. The fluorescence induction rise was examined repeatedly and detoxification kinetics were established from these data for the same position on the individual leaves. The herbicide‐dependent fluorescence rise decreased within hours in chlorotoluron‐tolerant but not in susceptible Triticum aestivum cultivars. In contrast, no significant reversion could be detected after metribuzin application in both tolerant and susceptible cultivars of Triticum durum. Near the fluorescence‐determined half‐inhibition of photosystem II, linear detoxification kinetics were obtained in individual leaves, thus providing an accurate measurement of relative detoxification rates.