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. The oxygen isotope composition ( δ18O ) ofleaf water ( δ18O leaf ) is an important determinant ofenvironmental and physiological information found in biological archives,but the system-scale understanding of the propagation of the δ18O of rain through soil and xylem water to δ18O leaf has not been verified for grassland. Here we report a unique andcomprehensive dataset of fortnightly δ18O observations in soil,stem and leaf waters made over seven growing seasons in a temperate,drought-prone, mixed-species grassland. Using the ecohydrology part of aphysically based, 18O -enabled soil–plant–atmosphere transfer model(MuSICA), we evaluated our ability to predict the dynamics of δ18O in soil water, the depth of water uptake, and the effects of soiland atmospheric moisture on 18 O enrichment of leaf water ( Δ18O leaf ) in this ecosystem. The model accurately predicted the δ18O dynamics of the different ecosystem water pools,suggesting that the model generated realistic predictions of the verticaldistribution of soil water and root water uptake dynamics. Observations andmodel predictions indicated that water uptake occurred predominantly fromshallow (  cm) soil depths throughout dry and wet periods in all years, presumably due (at least in part) to the effects of high grazingpressure on root system turnover and placement. Δ18O leaf responded to both soil and atmospheric moisture contents and was bestdescribed in terms of constant proportions of unenriched and evaporativelyenriched water (two-pool model). The good agreement between modelpredictions and observations is remarkable as model parameters describingthe relevant physical features or functional relationships of soil andvegetation were held constant with one single value for the entiremixed-species ecosystem.

The &lt;sup&gt;18&lt;/sup&gt;O ecohydrology of a grassland ecosystem – predictions and observations

The <sup>18</sup>O ecohydrology of a grassland ecosystem – predictions and observations