Multi-isotope reconstruction of Late Pleistocene large-herbivore biogeography and mobility patterns in Central Europe.

Interpretations of Late Pleistocene hominin adaptative capacities by archaeologists have focused heavily on their exploitation of certain prey and documented contemporary behaviours for these species. However, we cannot assume that animal prey-taxa ecology and ethology were the same in the past as in the present, or were constant over archaeological timescales. Sequential isotope analysis of herbivore teeth has emerged as a particularly powerful method of directly reconstructing diet, ecology and mobility patterns on sub-annual scales. Here, we apply 87Sr/86Sr isotope analysis, in combination with δ18O and δ13C isotope analysis, to sequentially sampled tooth enamel of prevalent herbivore species that populated Europe during the Last Glacial Period, including Rangifer tarandus, Equus sp. and Mammuthus primigenius. Our samples come from two open-air archaeological sites in Central Germany, Königsaue and Breitenbach, associated with Middle Palaeolithic and early Upper Palaeolithic cultures, respectively. We identify potential inter- and intra-species differences in range size and movement through time, contextualised through insights into diet and the wider environment. However, homogeneous bioavailable 87Sr/86Sr across large parts of the study region prevented the identification of specific migration routes. Finally, we discuss the possible influence of large-herbivore behaviour on hominin hunting decisions at the two sites.

Rapid oxygen isotopic exchange between bicarbonate and water during photosynthesis.

Whether rapid oxygen isotopic exchange between bicarbonate and water occurs in photosynthesis is the key to determine the source of oxygen by classic 18O-labeled photosynthetic oxygen evolution experiments. Here we show that both Microcystis aeruginosa and Chlamydomonas reinhardtii utilize a significant proportion (>16%) of added bicarbonate as a carbon source for photosynthesis. However, oxygen isotopic signal in added bicarbonate cannot be traced in the oxygen in organic matter synthesized by these photosynthetic organisms. This contradicts the current photosynthesis theory, which states that photosynthetic oxygen evolution comes only from water, and oxygen in photosynthetic organic matter comes only from carbon dioxide. We conclude that the photosynthetic organisms undergo rapid exchange of oxygen isotope between bicarbonate and water during photosynthesis. At the same time, this study also provides isotopic evidence for a new mechanism that half of the oxygen in photosynthetic oxygen evolution comes from bicarbonate photolysis and half comes from water photolysis, which provides a new explanation for the bicarbonate effect, and suggests that the Kok-Joliot cycle of photosynthetic oxygen evolution, must be modified to include a molecule of bicarbonate in addition to one molecule of water which in turn must be incorporated into the cycle instead of two water molecules. Furthermore, this study provides a theoretical basis for constructing a newer artificial photosynthetic reactor coupling light reactions with the dark reactions.

Different responses of oxygen and hydrogen isotopes in leaf and tree-ring organic matter to lethal soil drought.

The oxygen and hydrogen isotopic composition (δ18O, δ2H) of plant tissues are key tools for the reconstruction of hydrological and plant physiological processes and may therefore be used to disentangle the reasons for tree mortality. However, how both elements respond to soil drought conditions before death has rarely been investigated. To test this, we performed a greenhouse study and determined predisposing fertilization and lethal soil drought effects on δ18O and δ2H values of organic matter in leaves and tree rings of living and dead saplings of five European tree species. For mechanistic insights, we additionally measured isotopic (i.e. δ18O and δ2H values of leaf and twig water), physiological (i.e. leaf water potential and gas-exchange) and metabolic traits (i.e. leaf and stem non-structural carbohydrate concentration, carbon-to-nitrogen ratios). Across all species, lethal soil drought generally caused a homogenous 2H-enrichment in leaf and tree-ring organic matter, but a low and heterogenous δ18O response in the same tissues. Unlike δ18O values, δ2H values of tree-ring organic matter were correlated with those of leaf and twig water and with plant physiological traits across treatments and species. The 2H-enrichment in plant organic matter also went along with a decrease in stem starch concentrations under soil drought compared with well-watered conditions. In contrast, the predisposing fertilization had generally no significant effect on any tested isotopic, physiological and metabolic traits. We propose that the 2H-enrichment in the dead trees is related to (i) the plant water isotopic composition, (ii) metabolic processes shaping leaf non-structural carbohydrates, (iii) the use of carbon reserves for growth and (iv) species-specific physiological adjustments. The homogenous stress imprint on δ2H but not on δ18O suggests that the former could be used as a proxy to reconstruct soil droughts and underlying processes of tree mortality.

Advances in Oxygen Isotope Analysis of Phosphate by Electrospray Orbitrap Mass Spectrometry for Studying the Microbial Metabolism of Microorganisms.

Understanding the impact of human activities on the metabolic state of soil and aquatic environments is of paramount importance to implement measures for maintaining ecosystem services. Variations of natural abundance 18O/16O ratios in phosphate have been proposed as proxies for the holistic assessment of metabolic activity given the crucial importance of phosphoryl transfer reactions in fundamental biological processes. However, instrumental and procedural limitations inherent to oxygen isotope analysis in phosphate and organophosphorus compounds have so far limited the stable isotope-based evaluation of metabolic processes. Here, we discuss how recent developments in Orbitrap high resolution mass spectrometry enable measurements of 18O/16O ratios in phosphate and outline the critical mass spectrometry parameters for accurate and precise analysis. Subsequently, we evaluate the types of 18O kinetic isotope effects of phosphoryl transfer reactions and illustrate how novel analytical approaches will give rise to an improved understanding of 18O/16O ratio variations from biochemical processes affecting the microbial phosphorus metabolism.

Local circulation of elites punctuated by transregional mobility enabled steppe political consolidation in the Xiongnu nomadic state.

The Xiongnu polity (ca. 200 BC- 150 AD) emerged out of indigenous community-centered socio-political structures to forge a powerful state that commanded the Mongolian steppe and beyond. Underpinned by a highly mobile pastoralist population, accustomed to seasonally rhythmic moves and embedded in an equestrian culture that facilitated rapid transport over long-distances, it remains unclear precisely how the movement of commoners, local aristocrats and regional elites abetted the formation and organization of Xiongnu state structures. Here, we evaluate Xiongnu movement and dietary intake through multi-stable isotopic analyses of tooth enamel from directly dated Xiongnu intermediate elites recovered from the mortuary center of Baga Gazaryn Chuluu-a prominent granite outcrop set in the Gobi Desert. Carbon isotope (δ13C) analysis indicates millet was consumed by some individuals, but whether or not this C4 cultivar contributed to the diets of most elites remains ambiguous in this C3/C4 desert-steppe environment. The effectiveness of oxygen isotopes (δ18O) to establish mobility appears much reduced in steppe environments, where geospatially sensitive information appears disrupted by extraordinary seasonality in meteoric water oxygen isotopes, pronounced oxygen isotopic variation in potential drinking water sources, and culturally mediated drinking practices. Most revealing, strontium isotopes (87Sr/86Sr) indicate circulation of local elites around this central place and beyond, a mobility format that helped leaders cement their own position through political consolidation of spatially dispersed mobile pastoralist communities. The consistent presence at Baga Gazaryn Chuluu of extra-local intermediate elites also points toward the importance of transregional mobility in binding together the Xiongnu polity over the vast distances of the eastern steppe.

Clumped isotopes reveal relationship between mussel growth and river discharge.

Freshwater mussels preserve valuable information about hydrology, climate, and population dynamics, but developing seasonal chronologies can be problematic. Using clumped isotope thermometry, we produced high-resolution reconstructions of modern and historic (~ 1900) temperatures and δ18Owater from mussel shells collected from an impounded river, the Brazos in Texas, before and after damming. We also performed high-resolution growth band analyses to investigate relationships between mussel growth rate, rainfall, and seasonal temperature. Reconstructed δ18Owater and temperature vary little between the modern (3R5) and historic shell (H3R). However, a positive relationship between reconstructed δ18Owater and growth rate in H3R indicates that aside from diminished growth in winter, precipitation and flow rate are the strongest controls on mussel growth in both modern and pre-dam times. Overall, our results demonstrate (1) the impact, both positive and negative, of environmental factors such as flow alteration and temperature on mussel growth and (2) the potential for clumped isotopes in freshwater mussels as a paleohydrology and paleoclimate proxies in terrestrial environments.

Isotope Distribution Analysis in H218O Pulse-Labeled Trees Frozen with Liquid Nitrogen.

Tracer injection has long been recognized as a valuable tool for delineating tree hydraulics and assessing water transport pathways. Recently, isotope tracers have emerged as innovative instruments for investigating tree hydraulics, providing new insights into tree water dynamics. Nevertheless, there is a critical need for further research to comprehensively grasp water movement and distribution within trees. A previously introduced technique for analyzing the isotopic ratio of water in wet tissues, offering millimeter-scale resolution for visualizing tracer movement, faces challenges due to its underdeveloped sample preparation techniques. In this study, we introduced an H2 18O tracer into S. gracilistyla samples, exclusively comprising indeterminate roots, stems, and leaves, cultivated through hydroponics and grown within the current year. Our objective was to assess the axial distribution of the tracer in the xylem. Additionally, we devised a novel method for preparing frozen wet tissue samples, enhancing the repeatability and success rate of experiments. The results demonstrated that all frozen wet tissue samples exhibited an average water loss rate of less than 0.6%. Isotopic analysis of these samples unveiled a consistent decline in tracer concentration with increasing height in all Salix specimens, with three out of five samples revealing a significant isotope gradient. Our findings affirm the efficacy and practicality of combining isotopic labeling with freezing, stabilization, and preparation techniques. Looking ahead, our isotopic labeling and analysis methods are poised to transcend woody plants, finding extensive applications in plant physiology and ecohydrology.

Measurement of O2 Uptake and Evolution in Leaves In Vivo Using Stable Isotopes and Membrane Inlet Mass Spectrometry.

Oxygen is both product and substrate of photosynthesis and metabolism in plants, by oxygen evolution through water splitting and uptake by photorespiration and respiration. It is important to investigate these processes simultaneously in leaves, especially in response to environmental variables, such as light and temperature. To distinguish between processes that evolve or take up O2 in leaves in the light, in vivo gas exchange of stable isotopes of oxygen and membrane inlet mass spectrometry is used. A closed-cuvette system for gas exchange of leaf discs is described, using the stable isotopes 16O2 and 18O2, with a semi-permeable membrane gas inlet and isotope mass separation and detection by mass spectrometry. Measurement of evolution and uptake, as well as CO2 uptake, at a range of light levels allows composition of a light response curve, here described for French bean (Phaseolus vulgaris) and maize (Zea mays) leaf discs.

Effects of increasing atmospheric CO2 on leaf water δ18O values are small and are attenuated in grasses and amplified in dicotyledonous herbs and legumes when transferred to cellulose δ18O values.

The oxygen isotope composition of cellulose (δ18O values) has been suggested to contain information on stomatal conductance (gs) responses to rising pCO2. The extent by which pCO2 affects leaf water and cellulose δ18O values (δ18OLW and δ18OC) and the isotope processes that determine pCO2 effects on δ18OLW and δ18OC are, however, unknown. We tested the effects of pCO2 on gs, δ18OLW and δ18OC in a glasshouse experiment, where six plant species were grown under pCO2 ranging from 200 to 500 ppm. Increasing pCO2 caused a decline in gs and an increase in δ18OLW, as expected. Importantly, the effects of pCO2 on gs and δ18OLW were small and pCO2 effects on δ18OLW were not directly transferred to δ18OC but were attenuated in grasses and amplified in dicotyledonous herbs and legumes. This is likely because of functional group-specific pCO2 effects on the model parameter pxpex. Our study highlights important uncertainties when using δ18OC as a proxy for gs. Specifically, pCO2-triggered gs effects on δ18OLW and δ18OC are possibly too small to be detected in natural settings and a pCO2 effect on pxpex may render the commonly assumed negative linkage between δ18OC and gs to be incorrect, potentially confounding δ18OC based gs reconstructions.

Investigating isotopic markers for hazelnut geographical authentication: Promising variables and potential applications.

Hazelnuts' features and price are influenced by their geographical origin, making them susceptible to fraud, especially counterfeit claims regarding their provenance. Stable isotope analysis is a recognised approach to establish the geographical origin of foods, yet its potential in hazelnut authentication remains unexplored. In this prospective study, we assessed multiple isotopic markers in hazelnuts from different origins and evaluated the most promising variables for geographical authentication by chemometric tools. Our findings indicate that bulk δ18O, along with δ2H and δ13C in the main fatty acid methyl esters, exhibit significant potential in discriminating geographical origins, and 87Sr/86Sr analysis could serve as a proficient confirmatory tool. Though no single marker alone can differentiate between all the studied origins, employing a multi-isotopic approach based on PLS-DA models achieved up to 92.5 % accuracy in leave-10 %-out cross-validation. These findings will probably lay the groundwork for developing robust models for hazelnut geographical authentication based on larger datasets.

Tracing the impacts of ecological water replenishment on the sources and transformation of groundwater nitrate through isotope and microbial analysis.

Ecological water replenishment (EWR) changes the recharge conditions, flow fields, and physicochemical properties of regional groundwater. However, the resulting impacts on mechanisms regulating the sources and transformation of groundwater nitrate remain unclear. This study investigated how EWR influences the sources and transformation processes of groundwater nitrate using an integrated approach of Water chemistry analysis and stable isotopes (δ15N-NO3- and δ18O-NO3-) along with microbial techniques. The results showed that groundwater NO3-N decreased from 12.98 ± 7.39 mg/L to 7.04 ± 8.52 mg/L after EWR. Water chemistry and isotopic characterization suggested that groundwater nitrate mainly originated from sewage and manure. The Bayesian isotope mixing model (MixSIAR) indicated that EWR increased the average contribution of sewage and manure sources to groundwater nitrate from 46 % to 61 %, whereas that of sources of chemical fertilizer decreased from 43 % to 21 %. Microbial community analysis revealed that EWR resulted in a substantial decrease in the relative abundance of Pseudomonas spp denitrificans, from 13.7 % to 0.6 %. Both water chemistry and microbial analysis indicated that EWR weakened denitrification and enhanced nitrification in groundwater. EWR increases the contribution of nitrate to groundwater by promoting the release of sewage and feces in the unsaturated zone. However, the dilution effect caused by EWR was stronger than the contribution of sewage and fecal sources to groundwater nitrate. As a result, EWR helped to reduce groundwater nitrate concentrations. This study showed the effectiveness of integrated isotope and microbial techniques for delineating the sources and transformations of groundwater nitrate influenced by EWR.

The Pleistocene high-elevation environments between 2.02 and 0.6 Ma at Melka Kunture (Upper Awash Valley, Ethiopia) based upon stable isotope analysis.

Pleistocene environments are among the most studied issues in paleoecology and human evolution research in eastern Africa. Many data have been recorded from archaeological sites located at low and medium elevations (≤ 1500 m), whereas few contexts are known at 2000 m and above. Here, we present a substantial isotopic study from Melka Kunture, a complex of prehistoric sites located at 2000-2200 m above sea level in the central Ethiopian highlands. We analyzed the stable carbon and oxygen isotopic composition of 308 faunal tooth enamel samples from sites dated between 2.02 and 0.6 Ma to investigate the animal diets and habitats. The carbon isotopic results indicate that the analyzed taxa had C4-dominated and mixed C3-C4 diets with no significant diachronic changes in feeding behavior with time. This is consistent with faunal and phytolith analyses, which suggested environments characterized by open grasslands (with both C3 and C4 grasses), patches of bushes and thickets, and aquatic vegetation. However, palynological data previously documented mountain forests, woodlands, and high-elevation grasslands. Additionally, the carbon isotopic comparison with other eastern African localities shows that differences in elevation did not influence animal feeding strategies and habitat partitioning, even though plant species vary according to altitudinal gradients. In contrast, the oxygen isotopic comparison suggests significant differences consistent with the altitude effect. Our approach allows us to detect diverse aspects of animal behavior, habitat, and vegetation that should be considered when reconstructing past environments.

Isotopic biographies reveal horse rearing and trading networks in medieval London.

This paper reports a high-resolution isotopic study of medieval horse mobility, revealing their origins and in-life mobility both regionally and internationally. The animals were found in an unusual horse cemetery site found within the City of Westminster, London, England. Enamel strontium, oxygen, and carbon isotope analysis of 15 individuals provides information about likely place of birth, diet, and mobility during the first approximately 5 years of life. Results show that at least seven horses originated outside of Britain in relatively cold climates, potentially in Scandinavia or the Western Alps. Ancient DNA sexing data indicate no consistent sex-specific mobility patterning, although three of the five females came from exceptionally highly radiogenic regions. Another female with low mobility is suggested to be a sedentary broodmare. Our results provide direct and unprecedented evidence for a variety of horse movement and trading practices in the Middle Ages and highlight the importance of international trade in securing high-quality horses for medieval London elites.

Regional heterothermies recorded in the oxygen isotope composition of harbour seal skeletal elements.

Regional heterothermy is a strategy used by marine mammals to maintain a high and stable core body temperature, but its identification needs in situ measurements difficult to set up in extant wild organisms and inapplicable to extinct ones. We have analysed the oxygen isotope composition of bioapatite phosphate (δ18Op) from one permanent tooth and from thirty-six skeletal elements of one adult male harbour seal (Phoca vitulina) from the Baie de Somme (Hauts-de-France, France). We propose that the observed intra-skeletal δ18Op variability reflects tissue temperature heterogeneities typical of the pinniped regional heterothermy strategy. Our δ18Op data indicate that bone hydroxylapatite from harbour seal autopod skeletal elements (metacarpals, metatarsals, and phalanxes) mineralises at a lower temperature than that of the bone from the axial skeleton (e.g. vertebrae, ribs, and girdle bones). The results suggest that it is possible to locate a history of regional heterothermies in amphibious marine vertebrates using the δ18Op values of their mineralised tissues. This enables direct evaluation of the thermophysiology of both modern and fossil Pinnipedia from their skeletons opening perspectives on understanding their thermal adaptation to the marine environment in the fossil record. In addition to thermophysiology, oxygen isotope data from the permanent teeth of Pinnipedia, which are formed during the in utero phase from body fluid of the mother and at a stable temperature, could be valuable for locating the geographical areas inhabited by existing Pinnipedia females during their gestation period.

Half of the 18O enrichment of leaf sucrose is conserved in leaf cellulose of a C3 grass across atmospheric humidity and CO2 levels.

The 18O enrichment (Δ18O) of cellulose (Δ18OCel) is recognized as a unique archive of past climate and plant function. However, there is still uncertainty regarding the proportion of oxygen in cellulose (pex) that exchanges post-photosynthetically with medium water of cellulose synthesis. Particularly, recent research with C3 grasses demonstrated that the Δ18O of leaf sucrose (Δ18OSuc, the parent substrate for cellulose synthesis) can be much higher than predicted from daytime Δ18O of leaf water (Δ18OLW), which could alter conclusions on photosynthetic versus post-photosynthetic effects on Δ18OCel via pex. Here, we assessed pex in leaves of perennial ryegrass (Lolium perenne) grown at different atmospheric relative humidity (RH) and CO2 levels, by determinations of Δ18OCel in leaves, Δ18OLGDZW (the Δ18O of water in the leaf growth-and-differentiation zone) and both Δ18OSuc and Δ18OLW (adjusted for εbio, the biosynthetic fractionation between water and carbohydrates) as alternative proxies for the substrate for cellulose synthesis. Δ18OLGDZW was always close to irrigation water, and pex was similar (0.53 ± 0.02 SE) across environments when determinations were based on Δ18OSuc. Conversely, pex was erroneously and variably underestimated (range 0.02-0.44) when based on Δ18OLW. The photosynthetic signal fraction in Δ18OCel is much more constant than hitherto assumed, encouraging leaf physiological reconstructions.

Finding balance: Tree-ring isotopes differentiate between acclimation and stress-induced imbalance in a long-term irrigation experiment.

Scots pine (Pinus sylvestris L.) is a common European tree species, and understanding its acclimation to the rapidly changing climate through physiological, biochemical or structural adjustments is vital for predicting future growth. We investigated a long-term irrigation experiment at a naturally dry forest in Switzerland, comparing Scots pine trees that have been continuously irrigated for 17 years (irrigated) with those for which irrigation was interrupted after 10 years (stop) and non-irrigated trees (control), using tree growth, xylogenesis, wood anatomy, and carbon, oxygen and hydrogen stable isotope measurements in the water, sugars and cellulose of plant tissues. The dendrochronological analyses highlighted three distinct acclimation phases to the treatments: irrigated trees experienced (i) a significant growth increase in the first 4 years of treatment, (ii) high growth rates but with a declining trend in the following 8 years and finally (iii) a regression to pre-irrigation growth rates, suggesting the development of a new growth limitation (i.e. acclimation). The introduction of the stop treatment resulted in further growth reductions to below-control levels during the third phase. Irrigated trees showed longer growth periods and lower tree-ring δ13 C values, reflecting lower stomatal restrictions than control trees. Their strong tree-ring δ18 O and δ2 H (O-H) relationship reflected the hydrological signature similarly to the control. On the contrary, the stop trees had lower growth rates, conservative wood anatomical traits, and a weak O-H relationship, indicating a physiological imbalance. Tree vitality (identified by crown transparency) significantly modulated growth, wood anatomical traits and tree-ring δ13 C, with low-vitality trees of all treatments performing similarly regardless of water availability. We thus provide quantitative indicators for assessing physiological imbalance and tree acclimation after environmental stresses. We also show that tree vitality is crucial in shaping such responses. These findings are fundamental for the early assessment of ecosystem imbalances and decline under climate change.

Visible-Light-Driven Inactivation of Bacteria and H2 Generation Catalyzed by Oxygen-Vacancy-Rich One-Dimensional/Two-Dimensional W18O49/g-C3N4 Z-Scheme Heterostructures.

Z-scheme heterostructure-based photocatalysts consist of a reduction photocatalyst and an oxidation photocatalyst, enabling them to possess a high capacity for both reduction and oxidation. However, the coupling reaction between photocatalytic H2 generation through water reduction and sterilization using Z-scheme systems has been rarely reported. Herein, 1D W18O49 nanowires embedded over 2D g-C3N4 nanosheets are well-constructed as an integrated Z-scheme heterojunction. Experimental results and density functional theory calculations not only demonstrate the achievement of efficient interfacial charge separation and transport, leading to prolonged lifetime of photogenerated charge carriers, but also directly confirm the mechanism of Z-scheme charge transfer. As expected, the optimized W18O49/g-C3N4 nanostructure exhibits superior photocatalytic sterilization activity against Staphylococcus aureus as well as excellent H2 generation performance under visible-light irradiation (λ ≥ 420 nm). Due to its nontoxic nature, W18O49/g-C3N4 holds great potential in eradicating bacterial infections in living organisms.

Evaluation of Phosphorus Enrichment in Groundwater by Legacy Phosphorus in Orchard Soils with High Phosphorus Adsorption Capacity Using Phosphate Oxygen Isotope Analysis.

Long-term phosphorus (P) fertilization results in P accumulation in agricultural soil and increases the risk of P leaching into water bodies. However, evaluating P leaching into groundwater is challenging, especially in clay soil with a high P sorption capacity. This study examined whether the combination of PO4 oxygen isotope (δ18OPO4) analysis and the P saturation ratio (PSR) was useful to identify P enrichment mechanisms in groundwater. We investigated the groundwater and possible P sources in Kubi, western Japan, with intensive citrus cultivation. Shallow groundwater had oxic conditions with high PO4 concentrations, and orchard soil P accumulation was high compared with forest soil. Although the soil had a high P sorption capacity, the PSR was above the threshold, indicating a high risk of P leaching from the surface orchard soil. The shallow groundwater δ18OPO4 values were higher than the expected isotopic equilibrium with pyrophosphatase. The high PSR and δ18OPO4 orchard soil values indicated that P leaching from orchard soil was the major P enrichment mechanism. The Bayesian mixing model estimated that 76.6% of the P supplied from the orchard soil was recycled by microorganisms. This demonstrates the utility of δ18OPO4 and the PSR to evaluate the P source and biological recycling in groundwater.

Effects of particle size, storage conditions, and chemical pretreatments on carbon and oxygen isotopic measurements of modern tooth enamel.

Isotopic analysis of human tooth enamel can provide life history information useful in forensic identification. These applications depend on the availability of reference data documenting isotopic values for individuals with known life history and on the comparability of data from reference and case work samples. Here we build on previous methodological research, which has largely focused on paleontological and archaeological samples, and conduct experiments using enamel from modern human teeth targeting three sample preparation variables (sample particle size, storage conditions, and chemical pretreatments). Our results suggest that differences in particle size affect the efficiency of sample reactions during pretreatment and analysis, with coarse particles giving reduced loss of enamel carbonate during acid pretreatments but producing slightly higher oxygen isotope values than fine particles during analysis. Data for samples stored in dry and ambient environments following pretreatment were indistinguishable, suggesting no exchange of oxygen between samples and ambient water vapor. Finally, chemical pretreatments with a range of commonly used reactants and conditions showed a pervasive, moderate oxygen isotope shift associated with acetic acid treatment, which may be caused by exchange of enamel hydroxyl groups with reagents or rinse waters. Collectively, the results emphasize the importance of methodological standardization to improve comparability and reduce potential for bias in the forensic application of tooth enamel isotope data.

Quantifying isotope parameters associated with carbonyl-water oxygen exchange during sucrose translocation in tree phloem.

Stable oxygen isotope ratio of tree-ring α-cellulose (δ18Ocel) yields valuable information on many aspects of tree-climate interactions. However, our current understanding of the mechanistic controls on δ18Ocel is incomplete, with a knowledge gap existent regarding the fractionation effect characterizing carbonyl-water oxygen exchange during sucrose translocation from leaf to phloem. To address this insufficiency, we set up an experimental system integrating a vapor 18O-labeling feature to manipulate leaf-level isotopic signatures in tree saplings enclosed within whole-canopy gas-exchange cuvettes. We applied this experimental system to three different tree species to determine their respective relationships between 18O enrichment of sucrose in leaf lamina (Δ18Ol_suc) and petiole phloem (Δ18Ophl_suc) under environmentally/physiologically stable conditions. Based on the determined Δ18Ophl_suc-Δ18Ol_suc relationships, we estimated that on average, at least 25% of the oxygen atoms in sucrose undergo isotopic exchange with water along the leaf-to-phloem translocation path and that the biochemical fractionation factor accounting for such exchange is c. 34‰, markedly higher than the conventionally assumed value of 27‰. Our study represents a significant step toward quantitative elucidation of the oxygen isotope dynamics during sucrose translocation in trees. This has important implications with respect to improving the δ18Ocel model and its related applications in paleoclimatic and ecophysiological contexts.