Isotopic Composition Of Leaf Water Research Articles

Examination of the parameters controlling the triple oxygen isotope composition of grass leaf water and phytoliths at a Mediterranean site: a model–data approach

Abstract. Triple oxygen isotopes (17O-excess) of water are useful to trace evaporation at the soil–plant–atmosphere interface. The 17O-excess of plant silica, i.e., phytoliths, inherited from leaf water, was previously calibrated in growth chambers as a proxy of atmospheric relative humidity (RH). Here, using a model–data approach, we examine the parameters that control the triple oxygen isotope composition of bulk grass leaf water and phytoliths in natura, at the O3HP experimental platform located in the French Mediterranean area. A grass plot was equipped to measure for 1 year, all environmental and plant physiological parameters relevant for modeling the isotope composition of the grass leaf water. In particular, the triple oxygen and hydrogen isotope composition of atmospheric water vapor above the grass was measured continuously using a cavity ring-down spectrometer, and the grass leaf temperature was monitored at plot scale using an infrared (IR) radiometer. Grass leaves were collected in different seasons of the year and over a 24 h period in June. Grass leaf water was extracted by cryogenic vacuum distillation and analyzed by isotope ratio mass spectrometry (IRMS). Phytoliths were analyzed by IR–laser fluorination–IRMS after chemical extraction. We showed that the traditional Craig–Gordon steady-state model modified for grass leaves reliably predicts the triple oxygen isotope composition of leaf water during daytime but is sensitive to uncertainties on the leaf-to-air temperature difference. Deviations from isotope steady state at night are well represented in the triple oxygen isotope system and predictable by a non-steady-state model. The 17O-excess of phytoliths confirms the applicability of the 17O-excessphyto vs. RH equation established in previous growth chamber experiments. Further, it recorded average daytime RH over the growth period rather than daily RH, related to low transpiration and silicification during the night. This model–data approach highlights the utility of the triple oxygen isotope system to improve the understanding of water exchange at the soil–plant–atmosphere interface. The in natura experiment underlines the applicability of 17O-excess of phytoliths as a RH proxy.

Exploring the potential of Δ17O in CO2 for determining mesophyll conductance.

Mesophyll conductance to CO2 from the intercellular airspace to the CO2-H2O exchange site has been estimated using δ18O measurements (gm18). However, the gm18 estimates are affected by the uncertainties in the δ18O of leaf water where CO2-H2O exchange takes place and the degree of equilibration between CO2 and H2O. We show that measurements of Δ17O (i.e. Δ17O=δ17O-0.528×δ18O) can provide independent constraints on gm (gmΔ17) and that these gm estimates are less affected by fractionation processes during gas exchange. The gm calculations are applied to combined measurements of δ18O and Δ17O, and gas exchange in two C3 species, sunflower (Helianthus annuus L. cv "sunny") and ivy (Hedera hybernica L.), and the C4 species maize (Zea mays). The gm18 and gmΔ17--- estimates agree within the combined errors (p value 0.876). Both approaches are associated with large errors when the isotopic composition in the intercellular air space becomes close to the CO2-H2O exchange site. Although variations in Δ17O are low, it can be measured with much higher precision compared to δ18O. Measuring gmΔ17 has a few advantages compared to gm18: i) it is less sensitive to uncertainty in the isotopic composition of leaf water at the isotope exchange site and ii) the relative change in the gm due to an assumed error in the equilibration fraction θeq is lower for gmΔ17 compared to gm18. Thus, using Δ17O can complement and improve the gm estimates in settings where the δ18O of leaf water varies strongly, affecting the δ18O (CO2) difference between the intercellular air space and the CO2-H2O exchange site.

The effects on isotopic composition of leaf water and transpiration of adding a gas-exchange cuvette.

An expression was earlier derived for the non-steady state isotopic composition of a leaf when the composition of the water entering the leaf was not necessarily the same as that of the water being transpired (Farquhar and Cernusak 2005). This was relevant to natural conditions because the associated time constant is typically sufficiently long to ensure that the leaf water composition and fluxes of the isotopologues are rarely steady. With the advent of laser-based measurements of isotopologues, leaves have been enclosed in cuvettes and time courses of fluxes recorded. The enclosure modifies the time constant by effectively increasing the resistance to the one-way gross flux out of the stomata because transpiration increases the vapour concentration within the chamber. The resistance is increased from stomatal and boundary layer in series, to stomata, boundary layer and chamber resistance, where the latter is given by the ratio of leaf area to the flow rate out of the chamber. An apparent change in concept from one-way to net flux, introduced by Song, Simonin, Loucos and Barbour (2015) is resolved, and shown to be unnecessary, but the value of their data is reinforced.

Simulation of δ18O in Platycladus orientalis leaf water with different kinetic fractionation coefficients

Clarifying 18O isotope composition of leaf water (δL,b) would provide theoretical refe-rence for the study of leaf physiology and forest hydrology. We continuously monitored the concentration of atmospheric water vapor (Wa) and 18O isotope composition of atmospheric water vapor (δv) at the canopy of Platycladus lateralis plantation in the mountain area of Beijing. We analyzed the effects of kinetic fractionation coefficients Δk1(32%) and Δk2(28%) on the prediction of δL,b by combining the measured leaf water 18O isotope (δx) and δL,b of P. lateralis. The results showed that the diurnal variation of Wa was irregular. Atmospheric relative humidity (RH) showed a "V" shape of diurnal variation, and stomatal conductance (gs) increased first and then decreased at the diurnal scale. Wa, RH, and gs showed a significant negative correlation with δL,b when isotopes approached a steady-state equilibrium around noon. The kinetic fractionation coefficient Δk1 and Δk2 were applied to the Craig-Gordon model to predict δL,b under the isotopic quasi-steady-state condition. The results showed that the predicted values of Δk2 approached the observed values of δL,b. This result indicated that the application of Δk2 to the model was more consistent with the change of water isotope concentration in the leaves of P. lateralis in the mountain area of Beijing. These results would improve our understanding of water isotope enrichment model and evapotranspiration resolution model in leaves.

The <sup>18</sup>O ecohydrology of a grassland ecosystem – predictions and observations

Abstract. The oxygen isotope composition (δ18O) of leaf water (δ18Oleaf) is an important determinant of environmental and physiological information found in biological archives, but the system-scale understanding of the propagation of the δ18O of rain through soil and xylem water to δ18Oleaf has not been verified for grassland. Here we report a unique and comprehensive dataset of fortnightly δ18O observations in soil, stem and leaf waters made over seven growing seasons in a temperate, drought-prone, mixed-species grassland. Using the ecohydrology part of a physically based, 18O-enabled soil–plant–atmosphere transfer model (MuSICA), we evaluated our ability to predict the dynamics of δ18O in soil water, the depth of water uptake, and the effects of soil and atmospheric moisture on 18O enrichment of leaf water (Δ18Oleaf) in this ecosystem. The model accurately predicted the δ18O dynamics of the different ecosystem water pools, suggesting that the model generated realistic predictions of the vertical distribution of soil water and root water uptake dynamics. Observations and model predictions indicated that water uptake occurred predominantly from shallow (<20 cm) soil depths throughout dry and wet periods in all years, presumably due (at least in part) to the effects of high grazing pressure on root system turnover and placement. Δ18Oleaf responded to both soil and atmospheric moisture contents and was best described in terms of constant proportions of unenriched and evaporatively enriched water (two-pool model). The good agreement between model predictions and observations is remarkable as model parameters describing the relevant physical features or functional relationships of soil and vegetation were held constant with one single value for the entire mixed-species ecosystem.

Seasonal variation of δ18O and δ2H in leaf water of Fagus sylvatica L. and related water compartments

The study aims to assess variability in leaf water isotopic enrichment occurring in the field under natural conditions. We focused on seasonal variation and difference between sun-exposed and shaded leaves. Isotopic composition (δ18O, δ2H) of leaf water was monitored in a beech tree (Fagus sylvatica L.) growing in the forest-meadow ecotone together with δ18O (2H) of water compartments which are in close relation to this signal, namely twig and soil water. The sampling was carried out in approximately two-week intervals during five consecutive vegetation seasons.The δ18O (2H) data showed a distinct seasonal pattern and a consistency in relative differences between the seasons and sample categories. Leaf water was the most isotopically enriched water compartment. The leaf water enrichment decreased toward the autumn reflecting the change in δ18O (2H) of source water and evaporative demands. The soil and twig water isotopic signal was depleted against current precipitation as it partly retained the isotopic signature from winter precipitation however the seasonal pattern of soil and twig water followed that of precipitation. No significant differences between sun-exposed and shaded samples were detected. Nevertheless, the observed strong seasonal pattern of isotope composition of leaf, twig and soil water should be taken into account when using leaf water enrichment for further calculations or modeling.

Infidelity in the outback: climate signal recorded in Δ18O of leaf but not branch cellulose of eucalypts across an Australian aridity gradient.

The isotopic composition of leaf water in terrestrial plants is highly dependent upon a plant's environment. This isotopic signature can become integrated into organic molecules, allowing the isotopic composition of biomarkers such as cellulose to be used as sensitive paleo and climatic proxies. However, the mechanisms by which cellulose isotopic composition reflect environmental conditions are complex, and may vary between leaf and woody tissues. To date few empirical tests have been made on the relative roles of leaf-water enrichment and source water on the isotopic composition of leaf and wood cellulose within the same plant. Here, we study both leaf and branch wood cellulose, as well as xylem/source water of eucalypts across a 900 km aridity gradient in NE Australia. Across 11 sites, spanning average annual precipitation of 235-1400 mm and average relative humidity of 33-70%, we found a strong and consistent trend in leaf cellulose. However, once the effect of altered source water was considered we found wood cellulose to show no trend across this environmental gradient. We consider potential mechanisms that could explain the 'damping' of a climatic signal within wood cellulose and consider the implication and limitations on the use of tree-ring cellulose as a climate proxy.

Leaf water stable isotopes and water transport outside the xylem.

How water moves through leaves, and where the phase change from liquid to vapour occurs within leaves, remain largely mysterious. Some time ago, we suggested that the stable isotope composition of leaf water may contain information on transport pathways beyond the xylem, through differences in the development of gradients in enrichment within the various pathways. Subsequent testing of this suggestion provided ambiguous results and even questioned the existence of gradients in enrichment within the mesophyll. In this review, we bring together recent theoretical developments in understanding leaf water transport pathways and stable isotope theory to map a path for future work into understanding pathways of water transport and leaf water stable isotope composition. We emphasize the need for a spatially, anatomically and isotopically explicit model of leaf water transport.

Leaf water oxygen isotope measurement by direct equilibration

The oxygen isotope composition of leaf water imparts a signal to a range of molecules in the atmosphere and biosphere, but has been notoriously difficult to measure in studies requiring a large number of samples as a consequence of the labour-intensive extraction step. We tested a method of direct equilibration of water in fresh leaf samples with CO2 , and subsequent oxygen isotope analysis on an optical spectrometer. The oxygen isotope composition of leaf water measured by the direct equilibration technique was strongly linearly related to that of cryogenically extracted leaf water in paired samples for a wide range of species with differing anatomy, with an R(2) of 0.95. The somewhat more enriched values produced by the direct equilibration method may reflect lack of full equilibration with unenriched water in the vascular bundles, but the strong relationship across a wide range of species suggests that this difference can be adequately corrected for using a simple linear relationship.

Numerical modeling the isotopic composition of evapotranspiration in an arid artificial oasis cropland ecosystem with high-frequency water vapor isotope measurement

Stable isotopes of evapotranspiration (δET) provide us with useful information for tracing ecosystem processes in the study of biosphere–atmosphere interactions and hydrologic cycling. In this study, a numerical Iso-SPAC model with high frequency isotopic water vapor isotopic measurements was applied in an arid artificial oasis cropland ecosystem. The steady state assumption (SSA) and non-steady state (NSS) sub-models were tested for the isotopic composition of leaf water and transpiration flux. The modeled water/energy fluxes agreed well with eddy covariance observations, indicating that our model has the capacity to reproduce the observed water/energy flux behavior. Comparisons of modeled and estimated δET by the flux-gradient method using a cavity ring-down spectroscopy water vapor isotope analyzer as well as measured δL,b indicate that δT with SSA can reproduce better seasonal isotopic variations in ET flux and leaf water during the growing season. Uncertainties/errors for model output (δET, δE, δT and δL,b) under both SSA and NSS were discussed quantitatively. With corn growth, the isoflux of transpiration acted to increase δET and decrease enrichment in δL,b, inversely, the δET decreased and isotope enrichment in δL,b increased because of reduction of transpiration caused by frost occurred during 13th and 14th September. The results indicate that the Iso-SPAC model is a useful framework for simulating the dynamics of isotopic composition in the isotopic composition of plant transpiration and soil evaporation separately as well as for simultaneously integrating them for isotopic composition in the ET flux.

Partitioning evapotranspiration in a temperate grassland ecosystem: Numerical modeling with isotopic tracers

To partition evapotranspiration (ET) into soil evaporation (E) and vegetation transpiration (T), a new numerical Iso-SPAC (coupled heat, water with isotopic tracer in soil–plant–atmosphere-continuum) model was developed and applied to a temperate-grassland ecosystem in central Japan. Several models of varying complexity have been tested with the aim of obtaining the close to the true value for the isotope composition of leaf water and transpiration flux. The agreement between model predictions and observations demonstrated that the Iso-SPAC model with a steady-state assumption for transpiration flux can reproduce seasonal variations of all the surface energy balance components as well as isotope data (canopy foliage and ET flux). Uncertainties/errors for T/ET and each end members in assigned model parameters and measured input variables were discussed quantitatively. The T/ET increased with grass growth, and the sharp reduction caused by clear cutting was well reflected. The transpiration fraction ranged from 0.02 to 0.99 during growing seasons, and the mean value was 0.75 with a standard deviation of 0.24. Results indicate that SPAC model is a promising tool for integrating the isotopic fractionation processes with separate E and T in multispecies terrestrial ecosystems and has the advantage of enabling long-term assessment of ET partitioning as well as isotopic enrichment output at soil–plant–atmosphere interface. Our study reemphasize that isotope tracer approach is useful for evaluating quantitatively the relationships among hydrological components.

Multi-isotope labelling of organic matter by diffusion of <sup>2</sup>H/<sup>18</sup>O-H<sub>2</sub>O vapour and <sup>13</sup>C-CO<sub>2</sub> into the leaves and its distribution within the plant

Abstract. Isotope labelling is a powerful tool to study elemental cycling within terrestrial ecosystems. Here we describe a new multi-isotope technique to label organic matter (OM). We exposed poplars (Populus deltoides × nigra) for 14 days to an atmosphere enriched in 13CO2 and depleted in 2H218O. After 1 week, the water-soluble leaf OM (δ13C = 1346 ± 162‰) and the leaf water were strongly labelled (δ18O = −63 ± 8, δ2H = −156 ± 15‰). The leaf water isotopic composition was between the atmospheric and stem water, indicating a considerable back-diffusion of vapour into the leaves (58–69%) in the opposite direction to the net transpiration flow. The atomic ratios of the labels recovered (18O/13C, 2H/13C) were 2–4 times higher in leaves than in the stems and roots. This could be an indication of the synthesis of more condensed compounds in roots and stems (e.g. lignin vs. cellulose) or might be the result of O and H exchange and fractionation processes during phloem transport and biosynthesis. We demonstrate that the three major OM elements (C, O, H) can be labelled and traced simultaneously within the plant. This approach could be of interdisciplinary interest in the fields of plant physiology, palaeoclimatic reconstruction or soil science.

Effects of stomatal density and leaf water content on the ¹⁸O enrichment of leaf water.

Leaf water isotopic composition is imprinted in several biomarkers of interest and it is imperative that we understand the isotopic enrichment of leaf water. Here, we test the effect of stomatal density and leaf water content on the oxygen isotopic composition of leaf water in transgenic Arabidopsis plants expressing different stomatal densities, and several other species showing a range of stomatal density. We grew Arabidopsis plants hydroponically and collected other species in the field. Stomatal density and leaf water content were determined for each plant. We measured transpiration and extracted leaf water for isotopic determination. Using these measurements and the current leaf water isotope model, we calculated several of the parameters related to leaf water isotopic enrichment. High stomatal density promoted leaf water isotope enrichment. No conclusion, however, can be drawn regarding the effect of leaf water content on leaf water isotope enrichment. Factors such as transpiration might mask the effect of stomatal density on leaf water isotopic enrichment. We propose a method by which stomatal density can be incorporated in the current Peclet model of leaf water isotope enrichment. These findings have important applications in the use of plant-based metabolic proxies in paleoclimate studies.

A 220 ka terrestrial δ18O and deuterium excess biomarker record from an eolian permafrost paleosol sequence, NE-Siberia

The natural abundances of stable oxygen (18O/16O) and hydrogen isotopes (2H/1H) are valuable proxies of climate changes in the past. Yet, to date no continuous δ18O and only few δ2H records are available from loess–paleosol sequences. Taking advantage of a recently developed method based on compound-specific δ18O analyses of hemicellulose sugar biomarkers in soils (Zech and Glaser, 2009. RCM 23, 3522-3532), we here present a first terrestrial δ18O biomarker record from an eolian permafrost paleosol sequence in NE-Siberia that covers the last ~220ka.The δ18O values of the hemicellulose biomarkers arabinose and xylose range from 22.5 to 32.8‰ and from 21.3 to 31.9‰, respectively, and reveal systematic glacial–interglacial shifts. The modern topsoil and the interglacial paleosols exhibit more positive δ18O values, whereas the glacial paleosols are characterized by more negative δ18O values. This is in agreement with the δ2H record obtained for sedimentary n-alkane leaf wax biomarkers. We present a conceptual model for interpreting the combined δ18O and δ2H biomarker record. Based on this model, we suggest that both our δ18O and the δ2H records primarily reflect the temperature-controlled isotopic composition of paleoprecipitation modified by evaporative isotope enrichment of leaf water during transpiration.Considering fractionation factors during sugar and n-alkane biomarker biosynthesis allows reconstructing the leaf water isotopic composition and the deuterium excess of the leaf water. The deuterium excess may serve as proxy for evaporative enrichment and allows calculating relative humidity using a Craig–Gordon model. Accordingly, relative humidity in NE-Siberia was higher during marine isotope stage (MIS) 6 compared to MIS 2, 4 and 5d and thus could help explaining the much larger extent of the Late Saalian glaciation compared to the Weichselian glaciations.Using the Craig–Gordon model, we also calculated δ18O of the plant source water (δ18Osource water), which can be assumed to primarily reflect δ18O of paleoprecipitation. Our 220ka δ18Osource water record is well in agreement with the δ18Odiatom record from Crater Lake El'gygytgyn in NE-Siberia and enables a regional paleoclimate reconstruction and interpretation. Accordingly, summer temperature was periodically warmer than at present during the Weichselian glacial period and there is a strong July insolation forcing of the summer temperature in the extremely continental study area. Overall, our study highlights the great potential of the novel hemicellulose biomarker δ18O method for paleoclimate reconstructions, especially when combined with δ2H analyses of n-alkane lipid biomarkers.

Isotopic composition of transpiration and rates of change in leaf water isotopologue storage in response to environmental variables

During daylight hours, the isotope composition of leaf water generally approximates steady-state leaf water isotope enrichment model predictions. However, until very recently there was little direct confirmation that isotopic steady-state (ISS) transpiration in fact exists. Using isotope ratio infrared spectroscopy (IRIS) and leaf gas exchange systems we evaluated the isotope composition of transpiration and the rate of change in leaf water isotopologue storage (isostorage) when leaves were exposed to variable environments. In doing so, we developed a method for controlling the absolute humidity entering the gas exchange cuvette for a wide range of concentrations without changing the isotope composition of water vapour. The measurement system allowed estimation of (18)O enrichment both at the evaporation site and for bulk leaf water, in the steady state and the non-steady state. We show that non-steady-state effects dominate the transpiration isoflux even when leaves are at physiological steady state. Our results suggest that a variable environment likely prevents ISS transpiration from being achieved and that this effect may be exacerbated by lengthy leaf water turnover times due to high leaf water contents.

植物叶片水稳定同位素研究进展

植物叶片水稳定同位素研究进展

Temperature effect on leaf water deuterium enrichment and isotopic fractionation during leaf lipid biosynthesis: Results from controlled growth of C 3 and C 4 land plants

The hydrogen isotopic ratios ( 2H/ 1H) 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 2H 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 2H 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.

Diurnal variations of needle water isotopic ratios in two pine species

Diurnal fluctuations of leaf water isotope ratios (δ18O and δD) were measured for Jeffrey (Pinus jeffreyi Balf.) and lodgepole (Pinus contorta Douglas ex Louden) pine. Two trees per species were sampled every few hours on 15–16 October 2005 and 19–20 June 2006. Diurnal gas exchange was measured during the summer sampling. In fall 2005, leaf water δ18O ranged from 0.7 to 9.0‰, and leaf water δD ranged from −70 to −50‰. In summer 2006, leaf water δ18O ranged from 7.7 to 20.7‰, and leaf water δD ranged from −61 to −24‰. Diurnal variation of leaf water isotope values typically reached a maximum in early afternoon, began decreasing around midnight, and reached a minimum in mid-morning. Both periods showed a high degree of enrichment relative to source water, with leaf water–source water enrichments ranging up to 37.8‰ for δ18O, and up to 95‰ for δD. Leaf water enrichment varied by season with summer enrichment being greater than fall enrichment. A steady-state model (i.e., modified Craig–Gordon modeling) for leaf water isotope compositions did not provide a good fit to measured values of leaf water. In summer, a non-steady state model provided a better fit to the measured data than the steady-state model. Our findings demonstrate substantial leaf water enrichment above source water and diurnal variations in the isotopic composition of leaf water, which has application to understanding short-term variability of atmospheric gases (water vapor, CO2, O2), climate studies based on the isotopic composition of tree rings, and ecosystem water fluxes.

A modeling investigation of canopy‐air oxygen isotopic exchange of water vapor and carbon dioxide in a soybean field

The oxygen isotopes of CO2 and H2O (18O‐CO2 and 18O‐H2O) provide unique information regarding the contribution of terrestrial vegetation to the global CO2 and H2O cycles. In this paper, a simple isotopic land surface model was used to investigate processes controlling the isotopic exchange of 18O‐H2O and 18O‐CO2 between a soybean ecosystem and the atmosphere. We included in a standard land surface model a nonsteady state theory of leaf water isotopic composition, a canopy kinetic fractionation factor, and a big‐leaf parameterization of the 18O‐CO2 isoforcing on the atmosphere. Our model simulations showed that the Péclet effect was less important than the nonsteady state effect on the temporal dynamics of the water isotopic exchange. The model reproduced the highly significant and negative correlation between relative humidity and the ecosystem‐scale 18O‐CO2 isoforcing measured with eddy covariance. But the model‐predicted isoforcing was biased high in comparison to the observations. Model sensitivity analysis suggested that the CO2 hydration efficiency must have been much lower in the leaves of soybean in field conditions than previously reported. Understanding environmental controls on the hydration efficiency and the scaling from the leaf to the canopy represents an area in need of more research.

Controls on the D/H ratios of plant leaf waxes in an arid ecosystem

The extent to which leaf water D-enrichment (transpiration) and soil water D-enrichment (evaporation) affect the D/H ratio of plant leaf waxes remains a contentious issue, with important implications for paleohydrologic reconstructions. In this study we measure δD values of precipitation (δD_p), groundwater (δD_(gw)), plant xylem water (δD_(xw)) and leaf water (δD_(lw)) to understand their impact on the δD values of plant leaf wax n-alkanes (δD_(wax)) in an arid ecosystem. Our survey includes multiple species at four sites across an aridity gradient (80–30% relative humidity) in southern California. We find that many species take up groundwater or precipitation without significant fractionation. D-enriched soil water is a minor source even in species known to perform and utilize waters from hydraulic lift, such as Larrea tridentata (+10‰). Measurements of leaf water isotopic composition demonstrate that transpiration is an important mechanism for D-enrichment of leaf waters (+74 ± 20‰, 1σ), resulting in the smallest net fractionation yet reported between source water and leaf waxes (L. tridentata −41‰; multi-species mean value is −94 ± 21‰, 1σ). We find little change in leaf water D-enrichment or net fractionation across the climatic gradient sampled by our study, suggesting that a net fractionation of ca. −90‰ may be appropriate for paleohydrologic reconstructions in semi-arid to arid environments. Large interspecies offsets in net fractionations (1σ = 21‰) are potentially troublesome, given the observed floristic diversity and the likelihood of species assemblage changes with climate shifts.