On the progressive enrichment of the oxygen isotopic composition of water along a leaf

A model has been derived for the enrichment of heavy isotopes of water in leaves, including progressive enrichment along the leaf. In the model, lighter water is preferentially transpired leaving heavier water to diffuse back into the xylem and be carried further along the leaf. For this pattern to be pronounced, the ratio of advection to diffusion (Péclet number) has to be large in the longitudinal direction, and small in the radial direction. The progressive enrichment along the xylem is less than that occurring at the sites of evaporation in the mesophyll, depending on the isolation afforded by the radial Péclet number. There is an upper bound on enrichment, and effects of ground tissue associated with major veins are included. When transpiration rate is spatially nonuniform, averaging of enrichment occurs more naturally with transpiration weighting than with area‐based weighting. This gives zero average enrichment of transpired water, the modified Craig–Gordon equation for average enrichment at the sites of evaporation and the Farquhar and Lloyd (In Stable Isotopes and Plant Carbon‐Water Relations, pp. 47–70. Academic Press, New York, USA, 1993) prediction for mesophyll water. Earlier results on the isotopic composition of evolved oxygen and of retro‐diffused carbon dioxide are preserved if these processes vary in parallel with transpiration rate. Parallel variation should be indicated approximately by uniform carbon isotope discrimination across the leaf.     

Leaf vein fraction influences the Péclet effect and 18O enrichment in leaf water

The process of evaporation results in the fractionation of water isotopes such that the lighter ¹⁶O isotope preferentially escapes the gas phase leaving the heavier ¹⁸O isotope to accumulate at the sites of evaporation. This applies to transpiration from a leaf with the degree of fractionation dependent on a number of environmental and physiological factors that are well understood. Nevertheless, the ¹⁸O enrichment of bulk leaf water is often less than that predicted for the sites of evaporation. The advection of less enriched water in the transpiration stream has been suggested to limit the back diffusion of enriched evaporative site water (Péclet effect); however, evidence for this effect has been varied. In sampling across a range of species with different vein densities and saturated water contents, we demonstrate the importance of accounting for the relative ‘pool’ sizes of the vascular and mesophyll water for the interpretation of a Péclet effect. Further, we provide strong evidence for a Péclet signal within the xylem that if unaccounted for can lead to confounding of the estimated enrichment within the mesophyll water. This has important implications for understanding variation in the effective path length of the mesophyll and hence potentially the δ¹⁸O of organic matter.     

(18)O spatial patterns of vein xylem water,leaf water,and dry matter in cotton leaves

Three leaf water models (two-pool model, Péclet effect, and string-of-lakes) were assessed for their robustness in predicting leaf water enrichment and its spatial heterogeneity. This was achieved by studying the (18)O spatial patterns of vein xylem water, leaf water, and dry matter in cotton (Gossypium hirsutum) leaves grown at different humidities using new experimental approaches. Vein xylem water was collected from intact transpiring cotton leaves by pressurizing the roots in a pressure chamber, whereas the isotopic content of leaf water was determined without extracting it from fresh leaves with the aid of a purpose-designed leaf punch. Our results indicate that veins have a significant degree of lateral exchange with highly enriched leaf water. Vein xylem water is thus slightly, but progressively enriched in the direction of water flow. Leaf water enrichment is dependent on the relative distances from major veins, with water from the marginal and intercostal regions more enriched and that next to veins and near the leaf base more depleted than the Craig-Gordon modeled enrichment of water at the sites of evaporation. The spatial pattern of leaf water enrichment varies with humidity, as expected from the string-of-lakes model. This pattern is also reflected in leaf dry matter. All three models are realistic, but none could fully account for all of the facets of leaf water enrichment. Our findings acknowledge the presence of capacitance in the ground tissues of vein ribs and highlight the essential need to incorporate Péclet effects into the string-of-lakes model when applying it to leaves.     

Evaluation of models of leaf water 18O enrichment using measurements of spatial patterns of vein xylem water, leaf water and dry matter in maize leaves

The effectiveness of several leaf water models (‘string‐of‐lakes’, ‘desert river’ and the Farquhar–Gan model) are evaluated in predicting the enrichment of leaf water along a maize leaf at different humidities. Progressive enrichment of both vein xylem water and leaf water was observed along the blade. At the tip, the maximum observed enrichment for the vein water was 17.6‰ at 50% relative humidity (RH) whereas that for the leaf water was 50‰ at 34% RH and 19‰ at 75% RH. The observed leaf water maximum was a fraction (0.5–0.6) of the theoretically possible maximum. The ‘string‐of‐lakes’ and ‘desert river’ models predict well the variation of leaf water enrichment pattern with humidity but overestimate the average enrichment of bulk leaf water. However, the Farquhar–Gan model gives good prediction for these two aspects of leaf water enrichment. Using the anatomical dimensions of vein xylem overestimates the effective longitudinal Péclet number (P_l). Possible explanations for this discrepancy between the effective and the xylem‐based estimate of P_l are discussed. The need to characterize the heterogeneity of transpiration rate over the leaf surface in studies of leaf water enrichment is emphasized. The possibility that past atmospheric humidity can be predicted from the slope of the Δ¹⁸O spatial variation of leaf macrofossils found in middens is proposed.     

Non-steady-state, non-uniform transpiration rate and leaf anatomy effects on the progressive stable isotope enrichment of leaf water along monocot leaves

This study focuses on the spatial patterns of transpiration-driven water isotope enrichment (Delta(lw)) along monocot leaves. It has been suggested that these spatial patterns are the result of competing effects of advection and (back-)diffusion of water isotopes along leaf veins and in the mesophyll, but also reflect leaf geometry (e.g. leaf length, interveinal distance) and non-uniform gas-exchange parameters. We therefore developed a two-dimensional model of isotopic leaf water enrichment that incorporates new features, compared with previous models, such as radial diffusion in the xylem, longitudinal diffusion in the mesophyll, non-uniform gas-exchange parameters and non-steady-state effects. The model reproduces well all published measurements of Delta(lw) along monocot leaf blades, except at the leaf tip and given the uncertainties on measurements and model parameters. We show that the longitudinal diffusion in the mesophyll cannot explain the observed reduction in the isotope gradient at the leaf tip. Our results also suggest that the observed differences in Delta(lw) between C(3) and C(4) plants reflect more differences in mesophyll tortuosity rather than in leaf length or interveinal distance. Mesophyll tortuosity is by far the most sensitive parameter and different values are required for different experiments on the same plant species. Finally, using new measurements of non-steady-state, spatially varying leaf water enrichment we show that spatial patterns are in steady state around midday only, just as observed for bulk leaf water enrichment, but can be easily upscaled to the whole leaf level, regardless of their degree of heterogeneity along the leaf.     

Do pathways of water movement and leaf anatomical dimensions allow development of gradients in H218O between veins and the sites of evaporation within leaves?

The oxygen isotope enrichment of bulk leaf water (Δ_L) is often observed to be poorly predicted by the Craig–Gordon‐type models developed for evaporative enrichment from a body of water (Δ_e). The discrepancy between Δ_L and Δ_e may be explained by gradients in enrichment within the leaf as a result of convection of unenriched water to the sites of evaporation opposing the diffusion of enrichment away from the sites; a Péclet effect. However, this effect is difficult to quantify because the velocities of water movement within the leaf are unknown. This paper attempts to model the complex anatomy of a leaf, and hence such velocities, to assess if the gradients in H₂₁₈O required for a significant Péclet effect between the vein and the evaporation sites are possible within a leaf. Published dimensions of cells in wheat leaves are used to calculate the cross‐sectional areas perpendicular to the flow velocities of water through assumed pathways. By combining the ratio of actual to ‘slab’ velocities with anatomical lengths, equivalent lengths (L) emerge. In this way, it is concluded that if water moves only through the cell walls, or from cell to cell via either aquaporins or plasmodesmata, and evaporates from mesophyll cells, or the substomatal cells, or from the peristomatal region (a total of 15 combinations of assumptions), then the 15 central estimates of the values of L are between 9 and 200 mm. Each of these central estimates is subject to uncertainty, but overall their magnitude is important and estimates of L are comparable with those made from fitting to isotopic data (8 mm for wheat). It is concluded that significant gradients in enrichment between the vein and the evaporation sites are likely.     

Spatial variation of deuterium enrichment in bulk water of snowgum leaves

Deuterium enrichment of bulk water was measured and modeled in snowgum (Eucalyptus pauciflora Sieber ex Sprengel) leaves grown under contrasting air and soil humidity in arid and wet conditions in a glasshouse. A map of the enrichment was constructed with a resolution of 4 mm by using a newly designed cryodistillation method. There was progressively increasing enrichment in both longitudinal (along the leaf midrib) and transversal (perpendicular to the midrib) directions, most pronounced in the arid-grown leaf. The whole-leaf average of the enrichment was well below the value estimated by the Craig-Gordon model. The discrepancy between model and measurements persisted when the estimates were carried out separately for the leaf base and tip, which differed in temperature and stomatal conductance. The discrepancy was proportional to the transpiration rate, indicating the significance of diffusion-advection interplay (Péclet effect) of deuterium-containing water molecules in small veins close to the evaporating sites in the leaf. Combined Craig-Gordon and desert-river models, with or without the Péclet number, P, were used for predicting the leaf longitudinal enrichment. The predictions without P overestimated the measured values of deltadeuterium. Fixed P value partially improved the coincidence. We suggest that P should vary along the leaf length l to reconcile the modeled data with observations of longitudinal enrichment. Local values of P, P(l), integrating the upstream fraction of water used or the leaf area, substantially improved the model predictions.     

Evaporative enrichment and time lags between delta18O of leaf water and organic pools in a pine stand

Understanding ecosystem water fluxes has gained increasing attention, as climate scenarios predict a drier environment for many parts of the world. Evaporative enrichment of (18)O (Delta(18)O) of leaf water and subsequent enrichment of plant organic matter can be used to characterize environmental and physiological factors that control evaporation, based on a recently established mechanistic model. In a Pinus sylvestris forest, we measured the dynamics of oxygen isotopic composition (delta(18)O) every 6 h for 4 d in atmospheric water vapour, xylem sap, leaf water and water-soluble organic matter in current (N) and previous year (N-1) needles, phloem sap, together with leaf gas exchange for pooled N and N-1 needles, and relevant micrometeorological variables. Leaf water delta(18)O showed strong diel periodicity, while delta(18)O in atmospheric water vapour and in xylem sap showed little variation. The Delta(18)O was consistently lower for N than for N-1 needles, possibly related to phenological stage. Modelled leaf water Delta(18)O showed good agreement with measured values when applying a non-steady state evaporative enrichment model including a Péclet effect. We determined the time lags between delta(18)O signals from leaf water to water-soluble foliar organic matter and to phloem sap at different locations down the trunk, which clearly demonstrated the relevance of considering these time-lag effects for carbon transport, source-sink and carbon flux partitioning studies.     

The role of the mesophyll cell wall in leaf transpiration

Summary Evidence is presented which suggests that the mesophyll cell walls of cotton leaves may influence observed rates of transpiration.The net diffusive flux of water vapour, from the upper and lower surfaces of a leaf, was compared with the flux of nitrous oxide through a leaf and evidence obtained of an extra resistance in the water-vapour pathway associated with water transport in the mesophyll cell walls.This extra resistance appeared to be insignificant at low transpiration rates and in turgid leaves, but increased with transpiration rate and dehydration. The most likely explanation for its origin appeared to be a reduction in hydraulic conductivity across the internal cuticle which lines the outer surfaces of the mesophyll cell walls. In turn this served to reduce the relative vapour pressure at the sites of evaporation.The experiments were conducted under conditions where stomatal opening was induced by CO₂-free air. Under normal conditions stomatal closure would tend to reduce the development of this extra resistance. Even so, the results throw doubt on the validity of the long-standing assumption that the water-vapour pressure at the evaporation sites is equal to the saturation vapour pressure under all conditions.     

Modelling the isotope enrichment of leaf water

Farquhar and Gan [10] have proposed a model for the spatial variation in the isotopic enrichment of H₂¹⁸O across a leaf, which is specifically formulated for monocotyledoneous leaves. The model is based on the interaction between mass fluxes longitudinally within the xylem, and fluxes laterally through veinlets into the lamina mesophyll, where moisture leaves the leaf through transpiration. The lighter, more abundant, molecule H₂¹⁶O escapes preferentially with the evaporating water, resulting in the enrichment of H₂¹⁸O at these sites. Enriched water diffuses throughout the leaf, and it is this spatial distribution of enriched water which the model seeks to capture. In this paper we present a general formulation of the model in terms of mass flux, extending it to include variable transpiration rates across the leaf surface, as well as a tapering xylem. Solutions are developed for the general case and, since the solutions present in the form of Kummer functions, properties are established as well as methods for estimating the solutions under certain conditions relevant to the biology. The model output is compared with Gans data ([14, 15]) collected from maize plants.     

Temperature effect on leaf water deuterium enrichment and isotopic fractionation during leaf lipid biosynthesis: results from controlled growth of C3 and C4 land plants

The hydrogen isotopic ratios ((2)H/(1)H) of land plant leaf water and the carbon-bound hydrogen of leaf wax lipids are valuable indicators for climatic, physiological, metabolic and geochemical studies. Temperature will exert a profound effect on the stable isotopic composition of leaf water and leaf lipids as it directly influences the isotopic equilibrium (IE) during leaf water evaporation and cellular water dissociation. It is also expected to affect the kinetics of enzymes involved in lipid biosynthesis, and therefore the balance of hydrogen inputs along different biochemical routes. We conducted a controlled growth experiment to examine the effect of temperature on the stable hydrogen isotopic composition of leaf water and the biological and biochemical isotopic fractionations during lipid biosynthesis. We find that leaf water (2)H enrichment at 20°C is lower than that at 30°C. This is contrary to the expectation that at lower temperatures leaf water should be more enriched in (2)H due to a larger equilibrium isotope effect associated with evapotranspiration from the leaf if all other variables are held constant. A hypothesis is presented to explain the apparent discrepancy whereby lower temperature-induced down-regulation of available aquaporin water channels and/or partial closure of transmembrane water channel forces water flow to "detour" to a more convoluted apoplastic pathway, effectively increasing the length over which diffusion acts against advection as described by the Péclet effect (Farquhar and Lloyd, 1993) and decreasing the average leaf water enrichment. The impact of temperature on leaf water enrichment is not reflected in the biological isotopic fractionation or the biochemical isotopic fractionation during lipid biosynthesis. Neither the biological nor biochemical fractionations at 20°C are significantly different from that at 30°C, implying that temperature has a negligible effect on the isotopic fractionation during lipid biosynthesis.     

Variation in the oxygen isotope ratio of phloem sap sucrose from castor bean. Evidence in support of the Péclet effect

Theory suggests that the level of enrichment of (18)O above source water in plant organic material (Delta) may provide an integrative indicator of control of water loss. However, there are still gaps in our understanding of the processes affecting Delta. One such gap is the observed discrepancy between modeled enrichment of water at the sites of evaporation within the leaf and measured enrichment of the leaf water as a whole (Delta(L)). Farquhar and Lloyd (1993) suggested that this may be caused by a Péclet effect. It is also unclear whether organic material formed in the leaf reflects enrichment of water at the sites of evaporation within the leaf or Delta(L). To investigate this question castor bean (Ricinus communis L.) leaves, still attached to the plant, were sealed into a controlled-environment gas exchange chamber and subjected to a step change in leaf-to-air vapor pressure difference. Sucrose was collected from a cut on the petiole of the leaf in the chamber under equilibrium conditions and every hour for 6 h after the change in leaf-to-air vapor pressure difference. Oxygen isotope composition of sucrose in the phloem sap (Delta(suc)) reflected modeled Delta(L). A model is presented describing Delta(suc) at isotopic steady state, and accounts for 96% of variation in measured Delta(suc). The data strongly support the Péclet effect theory.     

The effect of water stress on photosynthesis and related parameters in Pinus halepensis

Net photosynthesis, transpiration, dark respiration rates and stomatal and mesophyll resistances were studied in young potted seedlings of Pinus halepensis Mill. under gradually decreasing soil and leaf water potentials. Stomatal resistance under non-limiting xylem water potentials was 6–7 times higher than mesophyll resistance. Stomata started to close at threshold xylem water potentials of −0.8 MPa, whereas mesophyll resistance started to increase at about −1.4 MPa. Decreasing xylem water potentials increased the CO₂ compensation point and decreased the water use efficiency (expressed by the photosynthesis to transpiration ratio) and dark respiration rate. It is concluded that at least part of the drought resistance characteristics of P. halepensis are associated with a sensitive stomatal mechanism which enables an efficient control of water loss.     

Modelling advection and diffusion of water isotopologues in leaves

We described advection and diffusion of water isotopologues in leaves in the non-steady state, applied specifically to amphistomatous leaves. This explains the isotopic enrichment of leaf water from the xylem to the mesophyll, and we showed how it relates to earlier models of leaf water enrichment in non-steady state. The effective length or tortuosity factor of isotopologue movement in leaves is unknown and, therefore, is a fitted parameter in the model. We compared the advection-diffusion model to previously published data sets for Lupinus angustifolius and Eucalyptus globulus. Night-time stomatal conductance was not measured in either data set and is therefore another fitted parameter. The model compared very well with the observations of bulk mesophyll water during the whole diel cycle. It compared well with the enrichment at the evaporative sites during the day but showed some deviations at night for E. globulus. It became clear from our analysis that night-time stomatal conductance should be measured in the future and that the temperature dependence of the tracer diffusivities should be accounted for. However, varying mesophyll water volume did not seem critical for obtaining a good prediction of leaf water enrichment, at least in our data sets. In addition, observations of single diurnal cycles do not seem to constrain the effective length that relates to the tortuosity of the water path in the mesophyll. Finally, we showed when simpler models of leaf water enrichment were suitable for applications of leaf water isotopes once weighted with the appropriate gas exchange flux. We showed that taking an unsuitable leaf water enrichment model could lead to large biases when cumulated over only 1 day.     

Cell water potential,osmotic potential,and turgor in the epidermis and mesophyll of transpiring leaves

Water potential, osmotic potential and turgor measurements obtained by using a cell pressure probe together with a nanoliter osmometer were compared with measurements obtained with an isopiestic psychrometer. Both types of measurements were conducted in the mature region of Tradescantia virginiana L. leaves under non-transpiring conditions in the dark, and gave similar values of all potentials. This finding indicates that the pressure probe and the osmometer provide accurate measurements of turgor, osmotic potentials and water potentials. Because the pressure probe does not require long equilibration times and can measure turgor of single cells in intact plants, the pressure probe together with the osmometer was used to determine in-situ cell water potentials, osmotic potentials and turgor of epidermal and mesophyll cells of transpiring leaves as functions of stomatal aperture and xylem water potential. When the xylem water potential was-0.1 MPa, the stomatal aperture was at its maximum, but turgor of both epidermal and mesophyll cells was relatively low. As the xylem water potential decreased, the stomatal aperture became gradually smaller, whereas turgor of both epidermal and mesophyll cells first increased and afterward decreased. Water potentials of the mesophyll cells were always lower than those of the epidermal cells. These findings indicate that evaporation of water is mainly occurring from mesophyll cells and that peristomatal transpiration could be less important than it has been proposed previously, although peristomatal transpiration may be directly related to regulation of turgor in the guard cells.     

Leaf water 18O and 2H maps show directional enrichment discrepancy in Colocasia esculenta

Spatial patterns of leaf water isotopes are challenging to predict because of the intricate link between vein and lamina water. Many models have attempted to predict these patterns, but to date, most have focused on monocots with parallel veins. These provide a simple system to study, but do not represent the majority of plant species. Here, a new protocol is developed using a Picarro induction module coupled to a cavity ringdown spectrometer to obtain maps of the leaf water isotopes (¹⁸O and ²H). The technique is applied to Colocasia esculenta leaves. The results are compared with isotope ratio mass spectrometry. In C. esculenta, a large enrichment in the radial direction is observed, but not in the longitudinal direction. The string‐of‐lakes model fails to predict the observed patterns, while the Farquhar–Gan model is more successful, especially when enrichment is accounted for along the radial direction. Our results show that reticulate‐veined leaves experience a larger enrichment along the axis of the secondary veins than along the midrib. We hypothesize that this is due to the lower major/minor vein ratio that leads to longer pathways between major veins and sites of evaporation.     

Ternary effects on the gas exchange of isotopologues of carbon dioxide

The ternary effects of transpiration rate on the rate of assimilation of carbon dioxide through stomata, and on the calculation of the intercellular concentration of carbon dioxide, are now included in standard gas exchange studies. However, the equations for carbon isotope discrimination and for the exchange of oxygen isotopologues of carbon dioxide ignore ternary effects. Here we introduce equations to take them into account. The ternary effect is greatest when the leaf-to-air vapour mole fraction difference is greatest, and its impact is greatest on parameters derived by difference, such as the mesophyll resistance to CO(2) assimilation, r(m) . We show that the mesophyll resistance to CO(2) assimilation has been underestimated in the past. The impact is also large when there is a large difference in isotopic composition between the CO(2) inside the leaf and that in the air. We show that this partially reconciles estimates of the oxygen isotopic composition of CO(2) in the chloroplast and mitochondria in the light and in the dark, with values close to equilibrium with the estimated oxygen isotopic composition of water at the sites of evaporation within the leaf.     

叶片水H218O富集的研究进展

 植物叶片水H₂¹⁸O富集对大气中O₂和CO₂的¹⁸O收支有着重要影响。蒸腾作用使植物叶片水H₂¹⁸O富集, 而植物叶片水H₂¹⁸O富集的程度主 要受大气水汽δ¹⁸O和植物蒸腾水汽δ¹⁸O的影响。过去, 通过引入稳态假设(蒸腾δ¹⁸O等于茎水δ¹⁸O)得到Craig-Gordon模型的闭合形式, 或 将植物整个叶片水δ¹⁸O经过Péclet效应校正后得到植物叶片水δ¹⁸O的富集程度。然而, 在几分钟到几小时的短时间尺度上, 植物叶片蒸腾 δ¹⁸O是变化的, 稳态假设是无法满足的。最近成功地实现了对大气水汽δ¹⁸O和δD的原位连续观测, 观测精度(小时尺度)可达到甚至优于稳定 同位素质谱仪的观测精度。在非破坏性条件下, 高时间分辨率和连续的大气水汽δ¹⁸O和蒸腾δ¹⁸O的动态观测, 将提高植物叶片水H₂¹⁸O富集的 预测能力。该文综述了植物叶片水H₂¹⁸O富集的理论研究的新进展、研究焦点和观测方法所存在的问题, 旨在进一步加深理解植物叶片水H₂¹⁸O 富集的过程及其机制。     

The enigma of effective path length for 18O enrichment in leaf water of conifers

The Péclet correction is often used to predict leaf evaporative enrichment and requires an estimate of effective path length (L). Studies to estimate L in conifer needles have produced unexpected patterns based on Péclet theory and leaf anatomy. We exposed seedlings of six conifer species to different vapour pressure deficits (VPD) in controlled climate chambers to produce steady‐state leaf water enrichment (in ¹⁸O). We measured leaf gas exchange, stable oxygen isotopic composition (δ¹⁸O) of input and plant waters as well as leaf anatomical characteristics. Variation in bulk needle water δ¹⁸O was strongly related to VPD. Conifer needles had large amounts of water within the vascular strand that was potentially unenriched (up to 40%). Both standard Craig–Gordon and Péclet models failed to accurately predict conifer leaf water δ¹⁸O without taking into consideration the unenriched water in the vascular strand and variable L. Although L was linearly related to mesophyll thickness, large within‐species variation prevented the development of generalizations that could allow a broader use of the Péclet effect in predictive models. Our results point to the importance of within needle water pools and isolating mechanisms that need further investigation in order to integrate Péclet corrections with ‘two compartment’ leaf water concepts.     

Effects of environmental parameters, leaf physiological properties and leaf water relations on leaf water delta18O enrichment in different Eucalyptus species

Stable oxygen isotope ratios (delta18O) have become a valuable tool in the plant and ecosystem sciences. The interpretation of delta18O values in plant material is, however, still complicated owing to the complex interactions among factors that influence leaf water enrichment. This study investigated the interplay among environmental parameters, leaf physiological properties and leaf water relations as drivers of the isotopic enrichment of leaf water across 17 Eucalyptus species growing in a common garden. We observed large differences in maximum daily leaf water delta18O across the 17 species. By fitting different leaf water models to these empirical data, we determined that differences in leaf water delta18O across species are largely explained by variation in the Péclet effect across species. Our analyses also revealed that species-specific differences in transpiration do not explain the observed differences in delta18O while the unconstrained fitting parameter 'effective path length' (L) was highly correlated with delta18O. None of the leaf morphological or leaf water related parameters we quantified in this study correlated with the L values we determined even though L was typically interpreted as a leaf morphological/anatomical property. A sensitivity analysis supported the importance of L for explaining the variability in leaf water delta18O across different species. Our investigation highlighted the importance of future studies to quantify the leaf properties that influence L. Obtaining such information will significantly improve our understanding of what ultimately determines the delta18O values of leaf water across different plant species.