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Riparian cottonwood forests in dry regions of western North America do not typically receive sufficient growing season precipitation to completely support their relatively high transpiration requirements. Water used in transpiration by riparian ecosystems must include alluvial groundwater or water stored in the potentially large reservoir of the unsaturated soil zone. We used the stable oxygen and hydrogen isotope composition of stem xylem water to evaluate water sources used by the dominant riparian cottonwood (Populus spp.) trees and shrubs (Shepherdia argentea and Symphoricarpos occidentalis) in Lethbridge, Alberta, during 3 years of contrasting environmental conditions. Cottonwoods did not exclusively take up alluvial groundwater but made extensive use of water sourced from the unsaturated soil zone. The oxygen and hydrogen isotope compositions of cottonwood stem water did not strongly overlap with those of alluvial groundwater, which were closely associated with the local meteoric water line. Instead, cottonwood stem water delta O-18 and delta H-2 values were located below the local meteoric water line, forming a line with a low slope that was indicative of water exposed to evaporative enrichment of heavy isotopes. In addition, cottonwood xylem water isotope compositions had negative values of deuterium excess (d-excess) and line-conditioned (deuterium) excess (lc-excess), both of which provided evidence that water taken up by the cottonwoods had been exposed to fractionation during evaporation. The shrub species had lower values of d-excess and lc-excess than had the cottonwood trees due to shallower rooting depths, and the d-excess values declined during the growing season, as shallow soil water that was taken up by the plants was exposed to increasing, cumulative evaporative enrichment. The apparent differences in functional rooting pattern between cottonwoods and the shrub species, strongly influenced the ratio of net photosynthesis to stomatal conductance (intrinsic water-use efficiency), as shown by variation among species in the delta C-13 values of leaf tissue.