Despite the utility of thermal imagery for characterising the impacts of water stress on plant physiology, few studies have been undertaken on plantation-grown conifers, including the most widely planted exotic species, radiata pine. Using data collected from a pot trial, where water was withheld from radiata pine over a nine-day period, the objectives of this study were to (i) determine how rapidly key physiological traits change in response to water stress and (ii) assess the utility of normalised canopy temperature, defined as canopy temperature–air temperature (Tc–Ta), for detecting these physiological changes. Volumetric water content remained high in the well-watered control treatment over the course of the experiment (0.47–0.48 m3 m−3) but declined rapidly in the water stress treatment from 0.47 m3 m−3 at 0 days after treatment (DAT) to 0.04 m3 m−3 at 9 DAT. There were no significant treatment differences in measurements taken at 0 DAT for Tc–Ta, stomatal conductance (gs), transpiration rate (E) or assimilation rate (A). However, by 1 DAT, differences between treatments in tree physiological traits were highly significant, and these differences continued diverging with values in the control treatment exceeding those of trees in the water stress treatment at 9 DAT by 42, 43 and 61%, respectively, for gs, E and A. The relationships between Tc–Ta and the three physiological traits were not significant at 0 DAT, but all three relationships were highly significant from as early as 1 DAT onwards. The strength of the relationships between Tc–Ta and the three physiological traits increased markedly over the duration of the water stress treatment, reaching a maximum coefficient of determination (R2) at 7 DAT when values were, respectively, 0.87, 0.86 and 0.67 for gs, E and A. The early detection of changes in tree physiology from 1 DAT onwards suggests that thermal imagery may be useful for a range of applications in field-grown radiata pine.