Field studies analyzing the stable isotope composition of xylem water are providing important information on ecosystem water relations. However, the capacity of stable isotopes to characterize the functioning of plants in their environment has not been fully explored because of methodological constraints on the extent and resolution at which samples could be collected and analysed. Here, we introduce an in situ method offering the potential to continuously monitor the stable isotope composition of tree xylem water via its vapour phase using a commercial laser-based isotope analyser and compact microporous probes installed into the xylem. Our technique enables efficient high-frequency measurement with intervals of only a few minutes per sample while eliminating the need for costly and cumbersome destructive collection of plant material and laboratory-based processing. We present field observations of xylem water hydrogen and oxygen isotope compositions obtained over several days including a labelled irrigation event and compare them against results from concurrent destructive sampling with cryogenic distillation and mass spectrometric analysis. The data demonstrate that temporal changes as well as spatial patterns of integration in xylem water isotope composition can be resolved through direct measurement. The new technique can therefore present a valuable tool to study the hydraulic architecture and water utilization of trees.

An experimental forest ecosystem drought Drought is affecting many of the world’ s forested ecosystems, but it has proved challenging to develop an ecosystem-level mechanistic understanding of the ways that drought affects carbon and water fluxes through forest ecosystems. Werner et al . used an experimental approach by imposing an artificial drought on an entire enclosed ecosystem: the Biosphere 2 Tropical Rainforest in Arizona (see the Perspective by Eisenhauer and Weigelt). The authors show that ecosystem-scale plant responses to drought depend on distinct plant functional groups, differing in their water-use strategies and their position in the forest canopy. The balance of these plant functional groups drives changes in carbon and water fluxes, as well as the release of volatile organic compounds into the atmosphere. —AMS

Forest water use has been difficult to quantify. One promising approach is to measure the isotopic composition of plant water, e.g., the transpired water vapor or xylem water. Because different water sources, e.g., groundwater versus shallow soil water, often show different isotopic signatures, isotopes can be used to investigate the depths from which plants take up their water and how this changes over time. Traditionally such measurements have relied on the extraction of wood samples, which provide limited time resolution at great expense, and risk possible artifacts. Utilizing a borehole drilled through a tree's stem, we propose a new method based on the notion that water vapor in a slow-moving airstream approaches isotopic equilibration with the much greater mass of liquid water in the xylem. We present two empirical data sets showing that the method can work in practice. We then present a theoretical model estimating equilibration times and exploring the limits at which the approach will fail. The method provides a simple, cheap, and accurate means of continuously estimating the isotopic composition of the source water for transpiration.

Abstract. The number of ecohydrological studies involving water stable isotope measurements has been increasing steadily due to technological (e.g., field-deployable laser spectroscopy and cheaper instruments) and methodological (i.e., tracer approaches or improvements in root water uptake models) advances in recent years. This enables researchers from a broad scientific background to incorporate water-isotope-based methods into their studies. Several isotope effects are currently not fully understood but might be essential when investigating root water uptake depths of vegetation and separating isotope processes in the soil–vegetation–atmosphere continuum. Different viewpoints exist on (i) extraction methods for soil and plant water and methodological artifacts potentially introduced by them, (ii) the pools of water (mobile vs. immobile) measured with those methods, and (iii) spatial variability and temporal dynamics of the water isotope composition of different compartments in terrestrial ecosystems. In situ methods have been proposed as an innovative and necessary way to address these issues and are required in order to disentangle isotope effects and take them into account when studying root water uptake depths of plants and for studying soil–plant–atmosphere interaction based on water stable isotopes. Herein, we review the current status of in situ measurements of water stable isotopes in soils and plants, point out current issues and highlight the potential for future research. Moreover, we put a strong focus and incorporate practical aspects into this review in order to provide a guideline for researchers with limited previous experience with in situ methods. We also include a section on opportunities for incorporating data obtained with described in situ methods into existing isotope-enabled ecohydrological models and provide examples illustrating potential benefits of doing so. Finally, we propose an integrated methodology for measuring both soil and plant water isotopes in situ when carrying out studies at the soil–vegetation–atmosphere continuum. Several authors have shown that reliable data can be generated in the field using in situ methods for measuring the soil water isotope composition. For transpiration, reliable methods also exist but are not common in ecohydrological field studies due to the required effort. Little attention has been paid to in situ xylem water isotope measurements. Research needs to focus on improving and further developing those methods. There is a need for a consistent and combined (soils and plants) methodology for ecohydrological studies. Such systems should be designed and adapted to the environment to be studied. We further conclude that many studies currently might not rely on in situ methods extensively because of the technical difficulty and existing methodological uncertainties. Future research needs to aim on developing a simplified approach that provides a reasonable trade-off between practicability and precision and accuracy.

Abstract. Stable isotopologues of water are widely used to derive relative root water uptake (RWU) profiles and average RWU depth in lignified plants. Uniform isotope composition of plant xylem water (δxyl) along the stem length of woody plants is a central assumption of the isotope tracing approach which has never been properly evaluated. Here we evaluate whether strong variation in δxyl within woody plants exists using empirical field observations from French Guiana, northwestern China, and Germany. In addition, supported by a mechanistic plant hydraulic model, we test hypotheses on how variation in δxyl can develop through the effects of diurnal variation in RWU, sap flux density, diffusion, and various other soil and plant parameters on the δxyl of woody plants. The hydrogen and oxygen isotope composition of plant xylem water shows strong temporal (i.e., sub-daily) and spatial (i.e., along the stem) variation ranging up to 25.2 ‰ and 6.8 ‰ for δ2H and δ18O, respectively, greatly exceeding the measurement error range in all evaluated datasets. Model explorations predict that significant δxyl variation could arise from diurnal RWU fluctuations and vertical soil water heterogeneity. Moreover, significant differences in δxyl emerge between individuals that differ only in sap flux densities or are monitored at different times or heights. This work shows a complex pattern of δxyl transport in the soil–root–xylem system which can be related to the dynamics of RWU by plants. These dynamics complicate the assessment of RWU when using stable water isotopologues but also open new opportunities to study drought responses to environmental drivers. We propose including the monitoring of sap flow and soil matric potential for more robust estimates of average RWU depth and expansion of attainable insights in plant drought strategies and responses.

Rationale: New methods to measure stable isotopes of soil and tree water directly in the field enable us to increase the temporal resolution of obtained data and advance our knowledge on the dynamics of soil and plant water fluxes. Only few field applications exist. However, these are needed to further improve novel methods and hence exploit their full potential. Methods:We tested the borehole equilibration method in the field and collected in situ and destructive samples of stable isotopes of soil, trunk and root xylem water over a 2.5-month experiment in a tropical dry forest under natural abundance conditions and following labelled irrigation. Water from destructive samples was extracted using cryogenic vacuum extraction. Isotope ratios were determined with IRIS instruments using cavity ring-down spectroscopy both in the field and in the laboratory.Results: In general, timelines of both methods agreed well for both soil and xylem samples. Irrigation labelled with heavy hydrogen isotopes clearly impacted the isotope composition of soil water and one of the two studied tree species. Intermethod deviations increased in consequence of labelling, which revealed their different capabilities to cover spatial and temporal heterogeneities. Conclusions:We applied the novel borehole equilibration method in a remote field location. Our experiment reinforced the potential of this in situ method for measuring xylem water isotopes in both tree trunks and roots and confirmed the reliability of gas permeable soil probes. However, in situ xylem measurements should be further developed to reduce the uncertainty within the range of natural abundance and hence enable their full potential. | INTRODUCTIONPlant transpiration fuels the hydrological cycle by returning 35% to 90% of water from land surfaces to the atmosphere. 1,2 Therefore, transpiration greatly influences our climate and impacts water availability in consequence of land use and climate change. 3,4 However, why is the quantification of this essential water flux so uncertain? This comes down to knowledge gaps in the mechanistic functioning of root water uptake (RWU) as well as plant rooting depth that persist despite its crucial role in predicting the future of one of our most important resources. 5 A major constraint is the practical difficulty in observing below-ground processes. Moreover, the magnitude and location of RWU are the results of multiple influencing factors, such as extent of the root system and its hydraulic properties,

Summary Root water uptake is a key ecohydrological process for which a physically based understanding has been developed in the past decades. However, due to methodological constraints, knowledge gaps remain about the plastic response of whole plant root systems to a rapidly changing environment. We designed a laboratory system for nondestructive monitoring of stable isotopic composition in plant transpiration of a herbaceous species (Centaurea jacea) and of soil water across depths, taking advantage of newly developed in situ methods. Daily root water uptake profiles were obtained using a statistical Bayesian multisource mixing model. Fast shifts in the isotopic composition of both soil and transpiration water could be observed with the setup and translated into dynamic and pronounced shifts of the root water uptake profile, even in well watered conditions. The incorporation of plant physiological and soil physical information into statistical modelling improved the model output. A simple exercise of water balance closure underlined the nonunique relationship between root water uptake profile on the one hand, and water content and root distribution profiles on the other, illustrating the continuous adaption of the plant water uptake as a function of its root hydraulic architecture and soil water availability during the experiment.

<p>Forest water use has been difficult to quantify. One promising approach is to measure the isotopic composition of plant water, e.g.<br>the transpired water vapor or xylem water, which often differs from that of other water vapor sources. Traditionally such<br>measurements have relied on the extraction of wood samples, which provide limited time resolution at great expense, and risk<br>possible artefacts. Utilizing a borehole drilled through a trees’ stem, we propose a new method based on the notion that water<br>vapor in a slow-moving airstream approaches equilibration with the much greater mass of liquid water in the xylem. We present<br>two empirical data sets showing that the method can work in practice. We then present theoretical models estimating the<br>equilibration times and exploring the limits at which the approach will fail. Given long enough boreholes and slow enough flows,<br>the method provides a simple, cheap, and accurate means of continuously estimating the isotopic composition of the source water<br>for transpiration.</p>