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Abstract Both theory and observations suggest that tree intrinsic water use efficiency (iWUE)—the ratio of photosynthetic carbon assimilation to stomatal conductance to water—increases with atmospheric CO₂. However, the strength of this relationship varies across sites and species, prompting questions about additional physiological constraints and environmental controls on iWUE. In this study, we analyzed tree core carbon isotope ratios to examine trends in, and drivers of, iWUE in 12 tree species common to the temperate forests of eastern North America, where forests have experienced changes in CO₂, climate, and atmospheric pollution in recent decades. Across all site-species combinations, we found that tree iWUE increased 22.3% between 1950 and 2011, coinciding with a 25.2% increase in atmospheric CO₂. iWUE trajectories varied markedly among tree functional groups and within species across sites. Needleleaf evergreen iWUE increased until circa 2002 before declining in recent years, while iWUE of broadleaf deciduous species continued to increase. The analysis of environmental controls on iWUE trends revealed smaller increases in iWUE in trees subjected to higher atmospheric pollution loads. Our results suggest that tree functional characteristics and atmospheric pollution history influence tree response to atmospheric CO₂, with implications for forest carbon and water balance in temperate regions.