Recent studies using water isotopes have shown that trees and streams appear to return distinct water pools to the hydrosphere. Cryogenically extracted plant and soil water isotopic signatures diverge from the meteoric water lines, suggesting that plants would preferentially use bound soil water, while mobile soil water that infiltrates the soil recharges groundwater and feeds streamflow all plots on meteoric water lines. These findings have been described under the “two water worlds” (TWW) hypothesis. In spite of growing evidence for the TWW hypothesis, several questions remain unsolved within the scope of this framework. Here, we address the TWW as a null hypothesis and further assess the following: (a) the theoretical biophysical feasibility for two distinct water pools to exist, (b) plant and soil processes that could explain the different isotopic composition between the two water pools, and (c) methodological issues that could explain the divergent isotopic signatures. Moreover, we propose a way forward under the framework of the TWW hypothesis, proposing alternative perspectives and explanations, experiments to further test them, and methodological advances that could help illuminate this quest. We further highlight the need to improve our sampling resolution of plants and soils across time and space. We ultimately propose a set of key priorities for future research to improve our understanding of the ecohydrological processes controlling water flows through the soil–plant‐atmosphere continuum.

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Abstract. Lake Chala (3 • 19 S, 37 • 42 E) is a steep-sided crater lake situated in equatorial East Africa, a tropical semiarid area with a bimodal rainfall pattern. Plants in this region are exposed to a prolonged dry season, and we investigated if (1) these plants show spatial variability and temporal shifts in their water source use; (2) seasonal differences in the isotopic composition of precipitation are reflected in xylem water; and (3) plant family, growth form, leaf phenology, habitat and season influence the xylem-to-leaf water deuterium enrichment. In this study, the δ 2 H and δ 18 O of precipitation, lake water, groundwater, plant xylem water and plant leaf water were measured across different plant species, seasons and plant habitats in the vicinity of Lake Chala. We found that plants rely mostly on water from the "short" rains falling from October to December (northeastern monsoon), as these recharge the soil after the long dry season. This plant-available, static water pool is only slightly replenished by the "long" rains falling from February to May (southeastern monsoon), in agreement with the "two water worlds" hypothesis, according to which plants rely on a static water pool while a mobile water pool recharges the groundwater. Spatial variability in water resource use exists in the study region, with plants at the lakeshore relying on a water source admixed with lake water. Leaf phenology does not affect water resource use. According to our results, plant species and their associated leaf phenology are the primary factors influencing the enrichment in deuterium from xylem water to leaf water (ε l/x ), with deciduous species giving the highest enrichment, while growth form and season have negligible effects. Our observations have important implications for the interpretation of δ 2 H of plant leaf wax n-alkanes (δ 2 H wax ) from paleohydrological records in tropical East Africa, given that the temporal variability in the isotopic composition of precipitation is not reflected in xylem water and that leaf water deuterium enrichment is a key factor in shaping δ 2 H wax . The large interspecies variability in xylem-leaf enrichment (24 ± 28 ‰) is potentially troublesome, taking into account the likelihood of changes in species assemblage with climate shifts.

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