Non-native plants have invaded over 100 millions of acres of western arid land in the US and dramatically altered nutrient cycling rates. Changes in water cycling caused by invasive species are of particular interest because primary production in the Western US is typically limited by water availability and aquifer recharge reflects plant demand. Large-scale invasions can, therefore, be expected to cause large-scale changes in hydrological cycles, but until recently, there have been considerable limitations in the ability to measure the timing, location, and extent of water use. Here we injected a tracer, deuterated water (D2O), into five soil depths in two sampling periods (May and June) in two adjacent plant communities (native and non-native dominated). Plants were sampled at several distances from the tracer addition area to determine the horizontal and vertical extent of water use in native and non-native communities. The tracer injection was coupled with measurements of leaf level stomatal conductance, leaf area index, and volumetric soil water content to estimate plant transpiration. We found that both native and non-native plants transpired water from primarily the top 60 cm of the soil (>75%), with a particular emphasis (≥ 50%) on shallow soil water (<10 cm) while lateral roots did not exceed 50 cm for most species. Higher leaf area index resulted in significantly more water being transpired from the native community. Some sharp distinctions in timing and location of tracer uptake resulting from the differing phenologies of the dominant species in each community were observed and confirmed previous mechanisms thought to govern plant assemblages in these communities. In May, the non-native community dominated by annual grasses had higher tracer uptake at 10 cm than the native community but began using deep water (higher tracer uptake at 80 cm) as annual grasses senesced and tap-rooted fobs became dominant in June. The perennial native species, however, used the entire soil profile from the moment they became active until they senesced. Our approach shows promise for overcoming the lack of resolution associated with natural abundance isotopes and other enrichment approaches, and for providing detailed measurements of plant water-use space.