Variability in weed control following pyrithiobac applications has been observed under field conditions. The influence of temperature on this variability was investigated. Results from field studies performed over two growing seasons identified plant and air temperatures at the time of herbicide treatment that correlated with whole-plant efficacy differences. Based on the field data, weed control with pyrithiobac was acceptable at application temperatures of 20 to 34 C. To investigate a potential source of thermal limitations on pyrithiobac efficacy, the thermal dependence of in vitro inhibition of acetolactate synthase (ALS), the site of action for pyrithiobac, was examined. A crude leaf extract of ALS was obtained from Amaranthus palmeri. Relative inhibitor potency (I50) values were obtained at saturating substrate conditions for temperatures from 10 to 50 C. Regression analysis of field activity against I50 values showed the two data sets to be highly correlated (R 2 = 0.88). The thermal dependence of enzyme/herbicide interactions may provide another means of understanding environmental factors limiting herbicidal efficacy and predicting herbicide inhibition at the whole-plant level.