Diurnal variation in the isotope composition of plant xylem water biases the depth of root-water uptake estimates

Abstract. 1. Stable isotopologues of water are a widely used tool to derive the depth of root water uptake (RWU) in lignified plants. Uniform isotope composition of plant xylem water (i-H2O-xyl) along the stem length is a central assumption, which has never been properly evaluated. 2. We studied the effects of diurnal variation in RWU, sap flux density and various other soil and plant parameters on i-H2O-xyl within a plant using a mechanistic plant hydraulic model and empirical field observations from French Guiana and northwestern China. 3. Our model predicts significant i-H2O-xyl variation arising from diurnal RWU fluctuations and vertical soil water heterogeneity. Moreover, significant differences in i-H2O-xyl emerge between individuals with different sap flux densities. In line with model predictions, field data show excessive i-H2O-xyl variation during the day or along stem length ranging up to 25.2 ‰ in δ2H and 6.8 ‰ in δ18O, largely exceeding the measurement error range. 4. Our work show that the fundamental assumption of uniform i-H2O-xyl is violated both theoretically and empirically and therefore a real danger exists of significant biases when using stable water isotopologues to assess RWU. We propose to include monitoring of sap flow and soil water potential for more robust RWU depth estimates.