Abstract. The stable isotopes of oxygen and hydrogen (δ²H and δ¹⁸O) have been widely used to investigate plant water source partitioning. These tracers have shed new light on patterns of plant water use in time and space. However, this black box approach has limited our source water interpretations and mechanistic understanding. Here, we combine measurements of stable isotope composition in xylem and soil water pools with measurements of plant hydraulics, fine root distribution and soil matric potential to investigate mechanism(s) driving tree water source partitioning. We used a 2 m³ lysimeter planted with a small willow tree (Salix viminalis) to conduct a high spatial-temporal resolution experiment. We found that tree water source partitioning was driven mainly by tree water status and not by patterns of fine root distribution. Source water partitioning was regulated by plant hydraulic response to changing atmospheric demand and soil matric potential. The depth distribution of soil matric potential appeared to be the largest control on the patterns of soil water partitioning during periods of tree water deficit. Contrary to the common steady state assumption in ecohydrological source water investigations, our results show that tree water use is a dynamic process, driven by tree water deficit. Overall, our findings suggest new research foci for future plant water isotopic investigations, highlighting the importance of hydrometric measurements from the plant perspective.