Leaf Waters Research Articles

Water taken up through the bark is detected in the transpiration stream in intact upper‐canopy branches

Alternative water uptake pathways through leaves and bark complement water supply with interception, fog or dew. Bark water-uptake contributes to embolism-repair, as demonstrated in cut branches. We tested whether bark water-uptake could also contribute to supplement xylem-water for transpiration. We applied bandages injected with 2 H-enriched water on intact upper-canopy branches of Pinus sylvestris and Fagus sylvatica in a boreal and in a temperate forest, in summer and winter, and monitored transpiration and online isotopic composition (δ2 H and δ18 O) of water vapour, before sampling for analyses of δ2 H and δ18 O in tissue waters. Xylem, bark and leaf waters from segments downstream from the bandages were 2 H-enriched whereas δ18 O was similar to controls. Transpiration was positively correlated with 2 H-enrichment. Isotopic compositions of transpiration and xylem water allowed us to calculate isotopic exchange through the bark via vapour exchange, which was negligible in comparison to estimated bark water-uptake, suggesting that water-uptake occurred via liquid phase. Results were consistent across species, forests and seasons, indicating that bark water-uptake may be more ubiquitous than previously considered. We suggest that water taken up through the bark could be incorporated into the transpiration stream, which could imply that sap-flow measurements underestimate transpiration when bark is wet.

Pronghorn (Antilocapra americana) enamel phosphate δ18O values reflect climate seasonality: Implications for paleoclimate reconstruction.

Stable oxygen isotope (δ18O) compositions from vertebrate tooth enamel are widely used as biogeochemical proxies for paleoclimate. However, the utility of enamel oxygen isotope values for environmental reconstruction varies among species. Herein, we evaluate the use of stable oxygen isotope compositions from pronghorn (Antilocapra americana Gray, 1866) enamel for reconstructing paleoclimate seasonality, an elusive but important parameter for understanding past ecosystems. We serially sampled the lower third molars of recent adult pronghorn from Wyoming for δ18O in phosphate (δ18OPO4) and compared patterns to interpolated and measured yearly variation in environmental waters as well as from sagebrush leaves, lakes, and rivers (δ18Ow). As expected, the oxygen isotope compositions of phosphate from pronghorn enamel are enriched in 18O relative to environmental waters. For a more direct comparison, we converted δ18Ow values into expected δ18OPO4* values (δ18OW‐PO4*). Pronghorn δ18OPO4 values from tooth enamel record nearly the full amplitude of seasonal variation from Wyoming δ18OW‐PO4* values. Furthermore, pronghorn enamel δ18OPO4 values are more similar to modeled δ18OW‐PO4* values from plant leaf waters than meteoric waters, suggesting that they obtain much of their water from evaporated plant waters. Collectively, our findings establish that seasonality in source water is reliably reflected in pronghorn enamel, providing the basis for exploring changes in the amplitude of seasonality of ancient climates. As a preliminary test, we sampled historical pronghorn specimens (1720 ± 100 AD), which show a mean decrease (a shift to lower values) of 1–2‰ in δ18OPO4 compared to the modern specimens. They also exhibit an increase in the δ18O amplitude, representing an increase in seasonality. We suggest that the cooler mean annual and summer temperatures typical of the 18th century, as well as enhanced periods of drought, drove differences among the modern and historical pronghorn, further establishing pronghorn enamel as excellent sources of paleoclimate proxy data.

Hydrologic Changes Drove the Late Miocene Expansion of C4 Grasslands on the Northern Indian Subcontinent

AbstractModern tropical and subtropical C4 grasslands and savannas were established during the late‐Miocene and Pliocene, over 20 Myr after evolutionary originations of the C4 photosynthetic pathway. This lag suggests environmental factors first limited and then favored C4 plants. Here, we examine the timing and drivers for the establishment of C4 grasslands on the Indian Subcontinent using carbon and hydrogen isotope signatures of plant‐wax n‐alkanes recovered from turbidites in the Bengal Fan. Like prior studies, we find C4 ecosystems in the Ganges‐Brahmaputra catchment first emerged at 7.4 Ma and subsequently expanded between 6.9 to ∼6.0 Ma. Hydrogen isotope values varied from 10.2 to 7.4 Ma and then increased after 7.4, which suggests intermittent drying began before the establishment of C4 grasslands with further drying at the onset of C4 expansion. Synthesis of published plant fossil data from the Siwalik Group of the Himalayan foreland basin documents an ecosystem trajectory from evergreen tropical forests to seasonally deciduous forests, and then expansive C4 grasslands. This trajectory coincided with a seasonally uneven drying trend due to both increased evaporation of plant leaf and soil waters and reduced rainfall, as identified in soil carbonate and tooth enamel data sets. Collectively the fossil, biomarker, and isotopic evidence reveal the development of modern C4 ecosystems on the Indian Subcontinent followed a series of ecosystem transformations driven by drying and fire feedbacks, and possibly declining atmospheric pCO2, beginning at 10.2 Ma and strengthening through the late Miocene.

Identification of bacteria from mangrove forest in Mamburungan, Tarakan City

Bacteria has a main role in the food chain and waters productivity. Bacteria in the mangrove ecosystem mostly are from litter, sediment, and sea waters. The research objective is to determine the genus of bacteria that was found in Avicennia marina and Sonneratia alba leaf litter as well as bacteria in waters and sediment from a mangrove forest in Mamburungan. The method was designed by using explorative descriptive, and the analysis parameter was the genus. This study was done in the field experimentally with two phases; they were collected of mangrove leaf litter, waters, and sediment, followed by identification of bacteria in the Fish quarantine station, quality control, and safety of Tarakan Class II Fisheries Laboratory. The results showed that seven genus bacteria of Sonneratia alba was found, such as Bacillus, Aeromonas, Listeria, Stapylococcus, Enterobacteria, Bacteroides, Actinobacillus and nine genus of Avicennia marina, they were Bacillus, Listeria, Aeromonas, Pseudomonas, Enterobacteria, Corynebacterium, Actinobacillus, Plesiomonas, Flavobacterium. In sediment, there were ten genus, i.e., Bacillus, Plesiomonas, Enterobacteria, Corynebacterium, Listeria, Aeromonas, Micrococcus, Actinobacillus, Bordetella parapertussis, Clostridium. In sea waters, there were eight genus like Bacillus, Listeria, Aeromonas, Corynebacterium, Micrococcus, Actinobacillus, Pseudomonas, and Enterobacteria. The dominant bacteria that were found in litter, sediment, and sea waters were Bacillus.

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.

Controls on leaf wax fractionation and δ2H values in tundra vascular plants from western Greenland

Hydrogen isotope ratios of leaf waxes are used to reconstruct past hydroclimate because they are a reflection of meteoric water, but the interpretation of these signatures from ancient sedimentary archives relies on a thorough understanding of the drivers of modern isotope variability and controls on fractionation. These studies are particularly valuable in the high latitudes, regions especially vulnerable to rapid climate change and increasingly used for plant-based proxy reconstructions of past hydroclimate, but also where modern vegetation is understudied compared to the lower latitudes. Here we investigate δ2H values from leaf wax n-alkanes of vascular tundra plants in the Kangerlussuaq area of western Greenland. We collected a variety of common tundra species to study possible interspecies variability in δ2H values including dwarf shrubs (Betula nana, Empetrum hermaphroditum, Salix glauca), forbs and graminoids (Vaccinium uliginosum, Rhododendron tomentosum, and Calamagrostis lapponica), a horsetail species (Equisetum arvense), and a submerged aquatic macrophyte from a local lake (Stuckenia filiformis). Using previously measured leaf and stem waters to help constrain potential drivers of leaf wax n-alkane δ2H values, we find that the overall net fractionation (εapp) from the studied species is −75 ± 20‰. The εapp at Kangerlussuaq is consistent with other studies of Arctic vegetation that find smaller εapp than from the majority of lower latitude sites. The fractionation of leaf water and xylem water (εlw/xw) and the fractionation of xylem water and precipitation (εxw/p) are both relatively constant, suggesting stable leaf and soil related fractionations across species. Estimates of biosynthetic fractionation (εbio), as evidenced from the fractionation of the δ2H values of n-alkanes and leaf water (εwax/lw), are not constant across species as sometimes assumed, and are small (average of εbio is −120 ± 27‰) compared to many published estimates. This supports a significant role in εbio shaping the εapp in this high latitude setting, where lipid biosynthesis may be driving differences in n-alkane δ2H values. This finding suggests that lipids in the Kangerlussuaq plants studied rely on the use of some proportion of different hydrogen sources during lipid synthesis, such as stored NADPH. The cumulative results of this survey of Kangerlussuaq area n-alkane δ2H values and water-wax fractionations suggest that fractionation in the high latitudes during the short summer growing season may play an important role in governing the small εapp compared to many low latitude sites. Better understanding of appropriate εapp and the importance of εbio in controlling plant wax fractionation from the high latitudes is necessary for future reconstructions of hydroclimate using leaf wax δ2H values in these regions.

Effect of continuous light on leaf wax isotope ratios in Betula nana and Eriophorum vaginatum: implications for Arctic paleoclimate reconstructions

Reconstructions of climate using leaf wax D/H ratios (δDwax) require accounting for the apparent isotopic fractionation (εapp) between plant source water and waxes. There have been conflicting publications on whether plants in the Arctic growing under 24-hour continuous light, fractionate less than temperate and tropical plants. In this study, we examine the effect of diurnal light (DL) versus 24-hour continuous light (CL) on the isotopic composition of leaf n-alkanes and n-acids in greenhouse experiments using two common Arctic plants (Eriophorum vaginatum, or tussock cottongrass and Betula nana, or dwarf birch). For E. vaginatum, the δDwax values of various wax homologues were 5–11‰ more positive for CL plants relative to their DL counterparts, whereas for B. nana, CL waxes were 3–24‰ more negative, suggesting that daylight length is not a unifying control on leaf wax D/H ratios of Arctic plants. The δ13Cwax of B. nana was more negative for plants grown in continuous light compared to diurnal light, reflecting lower water-use efficiency, associated with prolonged stomatal opening in the CL treatment. We modeled the impact of increasing stomatal conductance and effective flow path lengths (mimicking variable leaf morphologies) on the isotopic composition of leaf waters (δDlw) and find that variations in leaf-water enrichment may explain the variable δDwax responses seen between E. vaginatum and B. nana. We suggest that between-species differences in the δDlw response to light, and differences in the utilization of stored carbohydrates, were important for governing δDwax. Our greenhouse results suggest that Arctic plant leaf waxes do not consistently display reduced εapp values as a result of 24-hour day light, providing additional support for field observations.

Triple oxygen isotope composition of leaf waters in Mpala, central Kenya

Variations in triple oxygen isotopes have been used in studies of atmospheric photochemistry, global productivity and increasingly in studies of hydroclimate. Understanding the distribution of triple oxygen isotopes in plant waters is critical to studying the fluxes of oxygen isotopes between the atmosphere and hydrosphere, in which plants play an important role. In this paper we report triple oxygen isotope data for stem and leaf waters from Mpala, Kenya and explore how Δ17O, the deviation from an expected relationship between O17/O16 and O18/O16 ratios, in plant waters vary with respect to relative humidity and deuterium excess (d-excess). We observe significant variation in Δ17O among waters in leaves and stems from a single plant (up to 0.16‰ range in Δ17O in leaf water in a plant over the course of a signal day), which correlates to changes in relative humidity. A steady state model for evaporation in leaf water reproduces the majority of variation in Δ17O and d-excess we observed in leaf waters, except for samples that were collected in the morning, when relative humidity is high and the degree of fractionation in the system is minimal. The data and the steady state model indicate that the slope, λtransp, that links δ17O and δ18O values of stem and leaf waters and characterizes the fractionation during transpiration, is strongly influenced by the isotopic composition of ambient vapor when relative humidity is high. We observe a strong, positive relationship between d-excess and Δ17O, with a slope 2.2±0.2 per meg ‰−1, which is consistent with the observed relationship in tropical rainfall and in water in an evaporating open pan. The strong linear relationship between d-excess and Δ17O should be typical for any process involving evaporation or any other fractionation that is governed by kinetic effects.

Triple oxygen isotopes in biogenic and sedimentary carbonates

The 17O anomaly (Δ17O) of natural waters has been shown to be sensitive to evaporation in a way analogous to deuterium excess, with evaporated bodies of water (e.g., leaf waters, lake waters, animal body waters) tending to have lower Δ17O than primary meteoric waters. In animal body water, Δ17O relates to the intake of evaporated waters, evaporative effluxes of water, and the Δ17O value of atmospheric O2, which itself carries signatures of global carbon cycling and photochemical reactions in the stratosphere. Carbonates have the potential to record the triple oxygen isotope compositions of parent waters, allowing reconstruction of past water compositions, but such investigations have awaited development of methods for high-precision measurement of Δ17O of carbonate. We describe optimized methods based on a sequential acid digestion/reduction/fluorination approach that yield Δ17O data with the high precision (∼0.010‰, 1σ) needed to resolve subtle environmental signals. We report the first high-precision Δ17O dataset for terrestrial carbonates, focusing on vertebrate biogenic carbonates and soil carbonates, but also including marine invertebrates and high-temperature carbonates. We determine apparent three-isotope fractionation factors between the O2 analyte derived from carbonate and the parent waters of the carbonate. These in combination with appropriate temperature estimates (from clumped isotope thermometry, or known or estimated body temperatures) are used to calculate the δ18O and Δ17O of parent waters. The clearest pattern to emerge is the strong 17O-depletion in avian, dinosaurian, and mammalian body water (from analyses of eggshell and tooth enamel) relative to meteoric waters, following expected influences of evaporated water (e.g., leaf water) and atmospheric O2 on vertebrate body water. Parent waters of the soil carbonates studied here have Δ17O values that are similar to or slightly lower than global precipitation. Our results suggest that Δ17O will have useful application to paleoenvironmental studies of continental environments where the effects of evaporation are important, and where vertebrate body water may record an isotopic signal of evaporated water sources and atmospheric oxygen.

Understanding 2H/1H systematics of leaf wax n-alkanes in coastal plants at Stiffkey saltmarsh, Norfolk, UK

Interpretation of sedimentary n-alkyl lipid δ2H data is complicated by a limited understanding of factors controlling interspecies variation in biomarker 2H/1H composition. To distinguish between the effects of interrelated environmental, physical and biochemical controls on the hydrogen isotope composition of n-alkyl lipids, we conducted linked δ2H analyses of soil water, xylem water, leaf water and n-alkanes from a range of C3 and C4 plants growing at a UK saltmarsh (i) across multiple sampling sites, (ii) throughout the 2012 growing season, and (iii) at different times of the day. Soil waters varied isotopically by up to 35‰ depending on marsh sub-environment, and exhibited site-specific seasonal shifts in δ2H up to a maximum of 31‰. Maximum interspecies variation in xylem water was 38‰, while leaf waters differed seasonally by a maximum of 29‰. Leaf wax n-alkane 2H/1H, however, consistently varied by over 100‰ throughout the 2012 growing season, resulting in an interspecies range in the εwax/leafwater values of −79‰ to –227‰. From the discrepancy in the magnitude of these isotopic differences, we conclude that mechanisms driving variation in the 2H/1H composition of leaf water, including (i) spatial changes in soil water 2H/1H, (ii) temporal changes in soil water 2H/1H, (iii) differences in xylem water 2H/1H, and (iv) differences in leaf water evaporative 2H-enrichment due to varied plant life forms, cannot explain the range of n-alkane δ2H values we observed. Results from this study suggests that accurate reconstructions of palaeoclimate regimes from sedimentary n-alkane δ2H require further research to constrain those biological mechanisms influencing species-specific differences in 2H/1H fractionation during lipid biosynthesis, in particular where plants have developed biochemical adaptations to water-stressed conditions. Understanding how these mechanisms interact with environmental conditions will be crucial to ensure accurate interpretation of hydrogen isotope signals from the geological record.

The nocturnal water cycle in an open‐canopy forest

The movement of moisture into, out‐of, and within forest ecosystems is modulated by feedbacks that stem from processes which couple plants, soil, and the atmosphere. While an understanding of these processes has been gleaned from Eddy Covariance techniques, the reliability of the method suffers at night because of weak turbulence. During the summer of 2011, continuous profiles of the isotopic composition (i.e., δ18O and δD) of water vapor and periodic measurements of soil, leaf, and precipitation pools were measured in an open‐canopy ponderosa pine forest in central Colorado to study within‐canopy nocturnal water cycling. The isotopic composition of the nocturnal water vapor varies significantly based on the relative contributions of the three major hydrological processes acting on the forest: dewfall, exchange of moisture between leaf waters and canopy vapor, and periodic mixing between the canopy and background air. Dewfall proved to be surprisingly common (∼30% of the nights) and detectable on both the surface and within the canopy through the isotopic measurements. While surface dew could be observed using leaf wetness and soil moisture sensors, dew in the foliage was only measurable through isotopic analysis of the vapor and often occurred even when no dew accumulated on the surface. Nocturnal moisture cycling plays a critical role in water availability in forest ecosystems through foliar absorption and transpiration, and assessing these dynamics, as done here, is necessary for fully characterizing the hydrological controls on terrestrial productivity.

Laser-based measurements of 18O/16O stable isotope ratios (δ18O) in wine samples

Wine counterfeiting is an international, multi-billion dollar issue, with some estimates suggesting that up to 5% of wines sold at auctions or secondary markets are fraudulent. Isotope ratio mass spectrometer (IRMS) measurements of the 18O/16O stable isotope ratio (δ18O) of water-in-wine have been used for wine authentication; however, these analyses are time-consuming and costly. In this preliminary study, off-axis integrated cavity output spectroscopy (OA-ICOS) is used to quantify δ18O in wines. This laser-based method has been extensively used to study water isotopes for hydrological and medical applications. Recently, the development of a spectral contaminant identifier (SCI) has extended the application of these OA-ICOS analyzers to contaminated water samples (eg, plant, soil, and leaf waters). Here, we utilize OA-ICOS with the SCI to characterize wine samples (9%–15% ethanol), and show that the laser-based instrument provides a δ18O measurement precision of ±0.07‰ (1σ) and agrees with IRMS to within ±0.63‰ (1σ). Moreover, by training the SCI on isotopically-characterized wines, the agreement with IRMS improves to within ±0.30‰ (1σ). The utility of the instrument is demonstrated by measuring watered and mixed wines. The method presented here can be readily extended to address other food authentication applications.

Leaf-wax n -alkanes record the plant–water environment at leaf flush

Leaf-wax n-alkanes (2)H/(1)H ratios are widely used as a proxy in climate reconstruction. Although the broad nature of the relationship between n-alkanes δ(2)H values and climate is appreciated, the quantitative details of the proxy remain elusive. To examine these details under natural environmental conditions, we studied a riparian broadleaf angiosperm species, Populus angustifolia, growing on water with a constant δ(2)H value and monitored the δ(2)H values of leaf-wax n-alkanes and of stem, leaf, stream, and atmospheric waters throughout the entire growing season. Here we found the δ(2)H values of leaf-wax n-alkanes recorded only a 2-wk period during leaf flush and did not vary for the 19 weeks thereafter when leaves remained active. We found δ(2)H values of leaf-wax n-alkanes of P. angustifolia record conditions earlier in the season rather than fully integrating the entire growing season. Using these data, we modeled precipitation δ(2)H values during the time of wax synthesis. We observed that the isotope ratios of this precipitation generally were (2)H-enriched compared with mean annual precipitation. This model provides a mechanistic basis of the often-observed (2)H-enrichment from the expected fractionation values in studies of broadleaf angiosperm leaf-wax δ(2)H. In addition, these findings may have implications for the spatial and temporal uses of n-alkane δ(2)H values in paleoapplications; when both plant community and growth form are known, this study allows the isolation of the precipitation dynamics of individual periods of the growing season.

Reducing and correcting for contamination of ecosystem water stable isotopes measured by isotope ratio infrared spectroscopy

Concern exists about the suitability of laser spectroscopic instruments for the measurement of the (18)O/(16)O and (2)H/(1)H values of liquid samples other than pure water. It is possible to derive erroneous isotope values due to optical interference by certain organic compounds, including some commonly present in ecosystem-derived samples such as leaf or soil waters. Here we investigated the reliability of wavelength-scanned cavity ring-down spectroscopy (CRDS) (18)O/(16)O and (2)H/(1)H measurements from a range of ecosystem-derived waters, through comparison with isotope ratio mass spectrometry (IRMS). We tested the residual of the spectral fit S(r) calculated by the CRDS instrument as a means to quantify the difference between the CRDS and IRMS δ-values. There was very good overall agreement between the CRDS and IRMS values for both isotopes, but differences of up to 2.3‰ (δ(18)O values) and 23‰ (δ(2)H values) were observed in leaf water extracts from Citrus limon and Alnus cordata. The S(r) statistic successfully detected contaminated samples. Treatment of Citrus leaf water with activated charcoal reduced, but did not eliminate, δ(2)H(CRDS) - δ(2)H(IRMS) linearly for the tested range of 0-20% charcoal. The effect of distillation temperature on the degree of contamination was large, particularly for δ(2)H values but variable, resulting in positive, negative or no correlation with distillation temperature. S(r) and δ(CRDS) - δ(IRMS) were highly correlated, in particular for δ(2)H values, across the range of samples that we tested, indicating the potential to use this relationship to correct the δ-values of contaminated plant water extracts. We also examined the sensitivity of the CRDS system to changes in the temperature of its operating environment. We found that temperature changes ≥4 °C for δ(18)O values and ≥10 °C for δ(2)H values resulted in errors larger than the CRDS precision for the respective isotopes and advise the use of such instruments only in sufficiently temperature-stabilised environments.

Hydrogen isotope ratios of leaf wax n-alkanes in grasses are insensitive to transpiration

We analyzed hydrogen isotope ratios of high-molecular weight n-alkanes ( δD l) and oxygen isotope ratios of α-cellulose ( δ 18O C) for C 3 and C 4 grasses grown in the field and in controlled-environment growth chambers. The relatively firm understanding of 18O-enrichment in leaf water and α-cellulose was used to elucidate fractionation patterns of δD l signatures. In the different relative humidity environments of the growth chambers, we observed clear and predictable effects of leaf-water enrichment on δ 18O C values. Using a Craig–Gordon model, we demonstrate that leaf water in the growth chamber grasses should have experienced significant D-enriched due to transpiration. Nonetheless, we found no effect of transpirational D-enrichment on the δD l values. In field samples, we saw clear evidence of enrichment (correlating with relative humidity of the field sites) in both δ 18O C and δD l. These seemingly contrasting results could be explained if leaf waxes are synthesized in an environment that is isotopically similar to water entering plant roots due to either temporal or spatial isolation from evaporatively enriched leaf waters. For grasses in the controlled environment, there was no enrichment of source water, whereas enrichment of grass source water via evaporation from soils and/or stems was likely for grass samples grown in the field. Based on these results, evaporation from soils and/or stems appears to affect δD l, but transpiration from leaves does not. Further evidence for this conclusion is found in modeling expected net evapotranspirational enrichment. A Craig–Gordon model applied to each of the field sites yields leaf water oxygen isotope ratios that can be used to accurately predict the observed δ 18O C values. In contrast, the calculated leaf water hydrogen isotope ratios are more enriched than what is required to predict observed δD l values. These calculations lend support to the conclusion that while δ 18O C reflects both soil evaporation and transpiration, δD l appears to only record evaporation from soils and/or stems. Therefore, the δD of n-alkanes can likely be used to reconstruct the δD of water entering a leaf, supporting the soil-enrichment model of Smith and Freeman (2006). In both the field and controlled studies, we found significant photosynthetic pathway effects on n-alkane δD suggesting that biochemical pathways or plant phylogeny have a greater effect on leaf wax δD than leaf-water enrichment in grasses.

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.

A compound-specific n-alkane δ13C and δD approach for assessing source and delivery processes of terrestrial organic matter within a forested watershed in northern Japan

Abstract We measured molecular distributions and compound-specific hydrogen (δD) and stable carbon isotopic ratios (δ13C) of mid- and long-chain n-alkanes in forest soils, wetland peats and lake sediments within the Dorokawa watershed, Hokkaido, Japan, to better understand sources and processes associate with delivery of terrestrial organic matter into the lake sediments. δ13C values of odd carbon numbered C23–C33 n-alkanes ranged from −37.2‰ to −31.5‰, while δD values of these alkanes showed a large degree of variability that ranged from −244‰ to −180‰. Molecular distributions in combination with stable carbon isotopic compositions indicate a large contribution of C3 trees as the main source of n-alkanes in forested soils whereas n-alkanes in wetland soil are exclusively derived from marsh grass and/or moss. We found that the n-alkane δD values are much higher in forest soils than wetland peat. The higher δD values in forest samples could be explained by the enrichment of deuterium in leaf and soil waters due to increased evapotranspiration in the forest or differences in physiology of source plants between wetland and forest. A δ13C vs. δD diagram of n-alkanes among forest, wetland and lake samples showed that C25–C31 n-alkanes deposited in lake sediments are mainly derived from tree leaves due to the preferential transport of the forest soil organic matter over the wetland or an increased contribution of atmospheric input of tree leaf wax in the offshore sites. This study demonstrates that compound-specific δD analysis provides a useful approach for better understanding source and transport of terrestrial biomarkers in a C3 plant-dominated catchment.

18O 13C 16O in Earth’s atmosphere

The chemistry and budgets of atmospheric gases are constrained by their bulk stable isotope compositions (e.g., δ 13C values), which are based on mixing ratios of isotopologues containing one rare isotope (e.g., 16O 13C 16O). Atmospheric gases also have isotopologues containing two or more rare isotopes (e.g., 18O 13C 16O). These species have unique physical and chemical properties and could help constrain origins of atmospheric gases and expand the scope of stable isotope geochemistry generally. We present the first measurements of the abundance of 18O 13C 16O from natural and synthetic sources, discuss the factors influencing its natural distribution and, as an example of its applied use, demonstrate how its abundance constrains the sources of CO 2 in the Los Angeles basin. The concentration of 18O 13C 16O in air can be explained as a combination of ca. 1 ‰ enrichment (relative to the abundance expected if C and O isotopes are randomly distributed among all possible isotopologues) due to enhanced thermodynamic stability of this isotopologue during isotopic exchange with leaf and surface waters, ca. 0.1 ‰ depletion due to diffusion through leaf stomata, and subtle (ca. 0.05 ‰) dilution by 18O 13C 16O-poor anthropogenic CO 2. Some air samples are slightly (ca. 0.05 ‰) lower in 18O 13C 16O than can be explained by these factors alone. Our results suggest that 18O 13C 16O abundances should vary by up to ca. 0.2 ‰ with latitude and season, and might have measurable sensitivities to stomatal conductances of land plants. We suggest the greatest use of Δ 47 measurements will be to “leverage” interpretation of the δ 18O of atmospheric CO 2.

A mechanistic model for interpretation of hydrogen and oxygen isotope ratios in tree-ring cellulose

A mechanistic model is presented to quantify both the physical and biochemical fractionation events associated with hydrogen and oxygen isotope ratios in tree-ring cellulose. The model predicts the isotope ratios of tree-rings, incorporating both humidity and source water environmental information. Components of the model include (1) hydrogen and oxygen isotope effects associated with leaf water enrichment; (2) incorporation of leaf water isotope ratio values into photosynthetic carbohydrates along with the biochemical fractionation associated with autotrophic synthesis; (3) transport of exported carbohydrates (such as sucrose) from leaves to developing xylem in shoots and stems where cellulose is formed; (4) a partial exchange of oxygen and hydrogen isotopes in carbohydrates with xylem sap water during conversion into cellulose; and (5) a biochemical fractionation associated with cellulose synthesis. A modified version of the Craig–Gordon model for evaporative enrichment adequately described leaf water δD and δ 18O values. The leaf water model was robust over a wide range of leaf waters for both controlled experiments and field studies, far exceeding the range of values to be expected under natural conditions. The isotopic composition of cellulose was modeled using heterotrophic and autotrophic fractionation factors from the literature as well as the experimentally derived proportions of H and O that undergo exchange with xylem water during cellulose synthesis in xylem cells of tree-rings. The fraction of H and O from carbohydrates that exchange with xylem sap water was estimated to be 0.36 and 0.42, respectively. The proportions were based on controlled, long-term greenhouse experiments and field studies where the variations in the δD and δ 18O of tree-ring cellulose were measured under different source water isotopic compositions. The model prediction that tree-ring cellulose contains information on environmental water source and atmospheric vapor pressure deficit (related to relative humidity) was tested under both field and greenhouse conditions. This model was compared to existing models to explain cellulose isotope ratios under a wide range of source water and humidity conditions. Predictions from our model were consistent with observations, whereas other models showed large discrepancies as soon as the isotope ratios of source water and atmospheric water deviated from each other. Our model resolves the apparently conflicting and disparate interpretations of several previous cellulose stable isotope ratio studies.

Observations of hydrogen and oxygen isotopes in leaf water confirm the craig-gordon model under wide-ranging environmental conditions

The Craig-Gordon evaporative enrichment model of the hydrogen (deltaD) and oxygen (delta(18)O) isotopes of water was tested in a controlled-environment gas exchange cuvette over a wide range (400 per thousand deltaD and 40 per thousand delta(18)O) of leaf waters. (Throughout this paper we use the term "leaf water" to describe the site of evaporation, which should not be confused with "bulk leaf water" a term used exclusively for uncorrected measurements obtained from whole leaf water extractions.) Regardless of how the isotopic composition of leaf water was achieved (i.e. by changes in source water, atmospheric vapor deltaD or delta(18)O, vapor pressure gradients, or combinations of all three), a modified version of the Craig-Gordon model was shown to be sound in its ability to predict the deltaD and delta(18)O values of water at the site of evaporation. The isotopic composition of atmospheric vapor was shown to have profound effects on the deltaD and delta(18)O of leaf water and its influence was dependent on vapor pressure gradients. These results have implications for conditions in which the isotopic composition of atmospheric vapor is not in equilibrium with source water, such as experimental systems that grow plants under isotopically enriched water regimes. The assumptions of steady state were also tested and found not to be a major limitation for the utilization of the leaf water model under relatively stable environmental conditions. After a major perturbation in the deltaD and delta(18)O of atmospheric vapor, the leaf reached steady state in approximately 2 h, depending on vapor pressure gradients. Following a step change in source water, the leaf achieved steady state in 24 h, with the vast majority of changes occurring in the first 3 h. Therefore, the Craig-Gordon model is a useful tool for understanding the environmental factors that influence the hydrogen and oxygen isotopic composition of leaf water as well as the organic matter derived from leaf water.