Search

Unravelling the complexities of transpiration can be assisted by understanding the oxygen isotope composition of transpired water vapour (δE). It is often assumed that δE is at steady state, thereby mirroring the oxygen isotope composition of source water (δsource), but this assumption has never been tested at the whole-tree scale. This study utilized the unique infrastructure of 12 whole-tree chambers enclosing Eucalyptus parramattensis E.C.Hall trees to measure δE along with concurrent temperature and gas exchange data. Six chambers tracked ambient air temperature and six were exposed to an ambient +3 °C warming treatment. Day time means for δE were within 1.2‰ of δsource (-3.3‰) but varied considerably throughout the day. Our observations show that E. parramattensis trees are seldom transpiring at isotopic steady state over a diel period, but transpiration approaches source water isotopic composition over longer time periods., (© The Author(s) 2024. Published by Oxford University Press.)

Increases in temperatures and atmospheric CO2 concentration influence the growth performance of trees worldwide. The direction and intensity of tree growth and physiological responses to changing climate do, however, vary according to environmental conditions. Here we present complex, long-term, tree-physiological responses to unprecedented temperature increase in East Asia. For this purpose, we studied radial growth and isotopic (δ13C and δ18O) variations using tree-ring data for the past 100 yr of dominant Quercus mongolica trees from the cool-temperate forests from Hallasan, South Korea. Overall, we found that tree stem basal area increment, intercellular CO2 concentration and intrinsic water-use efficiency significantly increased over the last century. We observed, however, short-term variability in the trends of these variables among four periods identified by change point analysis. In comparison, δ18O did not show significant changes over time, suggesting no major hydrological changes in this precipitation-rich area. The strength and direction of growth-climate relationships also varied during the past 100 yr. Basal area increment (BAI) did not show significant relationships with the climate over the 1924-1949 and 1975-1999 periods. However, over 1950-1974, BAI was negatively affected by both temperature and precipitation, while after 2000, a temperature stimulus was observed. Finally, over the past two decades, the increase in Q. mongolica tree growth accelerated and was associated with high spring-summer temperatures and atmospheric CO2 concentrations and decreasing intrinsic water-use efficiency, δ18O and vapour pressure deficit, suggesting that the photosynthetic rate continued increasing under no water limitations. Our results indicate that the performance of dominant trees of one of the most widely distributed species in East Asia has benefited from recent global changes, mainly over the past two decades. Such findings are essential for projections of forest dynamics and carbon sequestration under climate change., (© The Author(s) 2024. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Disentangling the factors influencing the climate sensitivity of trees is crucial to understanding the susceptibility of forests to climate change. Reducing tree-to-tree competition and mixing tree species are two strategies often promoted to reduce the drought sensitivity of trees, but it is unclear how effective these measures are in different ecosystems. Here, we studied the growth and physiological responses to climate and severe droughts of silver fir and Douglas-fir growing in pure and mixed conditions at three sites in Switzerland. We used tree-ring width data and carbon (δ13C), oxygen (δ18O) and hydrogen (δ2H) stable isotope ratios from tree-ring cellulose to gain novel information on water relations and the physiology of trees in response to drought and how tree species mixture and competition modulate these responses. We found significant differences in isotope ratios between trees growing in pure and mixed conditions for the two species, although these differences varied between sites, e.g. trees growing in mixed conditions had higher δ13C values and tree-ring width than trees growing in pure conditions for two of the sites. For both species, differences between trees in pure and mixed conditions regarding their sensitivity to temperature, precipitation, climatic water balance and vapor pressure deficit were minor. Furthermore, trees growing in pure and mixed conditions showed similar responses of tree-ring width and isotope ratios to the past severe droughts of 2003, 2015 and 2018. Competition had only a significantly negative effect on δ13C of silver fir, which may suggest a decrease in photosynthesis due to higher competition for light and nutrients. Our study highlights that tree species mixture may have only moderate effects on the radial growth and physiological responses of silver fir and Douglas-fir to climatic conditions and that site condition effects may dominate over mixture effects., (© The Author(s) 2024. Published by Oxford University Press.)

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., (© The Author(s) 2024. Published by Oxford University Press.)

The stable hydrogen and oxygen isotope technique is typically used to explore plant water uptake; however, the accuracy of the technique has been challenged by hydrogen isotope offsets between plant xylem water and its potential source water. In this study, the soil hydrogen and oxygen isotope waterline was used to correct the hydrogen isotope offsets for Salix psammophila and Caragana korshinskii, two typical shrub species on the Chinese Loess Plateau. Five different types of isotopic data [(i) δ18O, (ii) δ2H, (iii) combination δ18O with δ2H, (iv) corrected δ2H and (v) combination δ18O with corrected δ2H] were separately used to determine the water-use patterns of the two shrubs. The δ2H offset values of S. psammophila and C. korshinskii did not show significant temporal variation among the sampling months (May, July and September) but showed notable differences between the two shrubs (-0.4 ± 0.5‰ in S. psammophila vs -4.3 ± 0.9‰ in C. korshinskii). The obtained water absorption proportion (WAP) of S. psammophila in the different soil layers (0-20, 20-60 and 60-200 cm) did not differ significantly among the five different input data types. However, compared with the input data types (iii) and (v), the data types (i), (ii) and (iv) overestimated the WAP of C. korshinskii in the 0-20 cm soil layer and underestimated that in the 60-200 cm layer. The data type (iii) overestimated the WAP of C. korshinskii in 0-20 cm soil layer (25.9 ± 0.8%) in July in comparison with the WAP calculated based on data type (v) (19.1 ± 1.1%). The combination of δ18O and corrected δ2H, i.e., data type (v), was identified as the best data type to determine the water use patterns of C. korshinskii due to the strong correlation between the calculated WAP and soil water content and soil sand content. In general, S. psammophila mainly used (57.9-62.1%) shallow soil water (0-60 cm), whereas C. korshinskii mainly absorbed (52.7-63.5%) deep soil water (60-200 cm). We confirm that the hydrogen isotope offsets can cause significant errors in determining plant water uptake of C. korshinskii, and provide valuable insights for accurately quantifying plant water uptake in the presence of hydrogen isotope offsets between xylem and source water. This study is significant for facilitating the application of the stable hydrogen and oxygen isotope technique worldwide, and for revealing the response mechanism of shrub key ecohydrological and physiological processes to the drought environment in similar climate regions., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Continuous cover forestry (CCF) has gained interest as an alternative to even-aged management particularly on drained peatland forests. However, relatively little is known about the physiological response of suppressed trees when larger trees are removed as a part of CCF practices. Consequently, studies concentrating on process-level modeling of the response of trees to selection harvesting are also rare. Here, we compared, modeled and measured harvest response of previously suppressed Norway spruce (Picea abies) trees to a selection harvest. We quantified the harvest response by collecting Norway spruce tree-ring samples in a drained peatland forest site and measuring the change in stable carbon and oxygen isotopic ratios of wood formed during 2010-20, including five post-harvest years. The measured isotopic ratios were compared with ecosystem-level process model predictions for ${\kern0em }^{13}$C discrimination and ${\kern0em }^{18}$O leaf water enrichment. We found that the model predicted similar but lower harvest response than the measurements. Furthermore, accounting for mesophyll conductance was important for capturing the variation in ${\kern0em }^{13}$C discrimination. In addition, we performed sensitivity analysis on the model, which suggests that the modeled ${\kern0em }^{13}$C discrimination is sensitive to parameters related to CO2 transport through stomata to the mesophyll., (© The Author(s) 2023. Published by Oxford University Press.)

Stable isotope ratio analysis of tree rings has been widely and successfully applied in recent decades for climatic and environmental reconstructions. These studies were mostly conducted at an annual resolution, considering one measurement per tree ring, often focusing on latewood. However, much more information could be retrieved with high-resolution intra-annual isotope studies, based on the fact that the wood cells and the corresponding organic matter are continuously laid down during the growing season. Such studies are still relatively rare, but have a unique potential for reconstructing seasonal climate variations or short-term changes in physiological plant properties, like water-use efficiency. The reason for this research gap is mostly technical, as on the one hand sub-annual, manual splitting of rings is very tedious, while on the other hand automated laser ablation for high-resolution analyses is not yet well established and available. Here, we give an update on the current status of laser ablation research for analysis of the carbon isotope ratio (δ13C) of wood, describe an easy-to-use laser ablation system, its operation and discuss practical issues related to tree core preparation, including cellulose extraction. The results show that routine analysis with up to 100 laser shot-derived δ13C-values daily and good precision and accuracy (ca. 0.1‰) comparable to conventional combustion in an elemental analyzer are possible. Measurements on resin-extracted wood is recommended as most efficient, but laser ablation is also possible on cellulose extracted wood pieces. Considering the straightforward sample preparation, the technique is therefore ripe for wide-spread application. With this work, we hope to stimulate future progress in the promising field of high-resolution environmental reconstruction using laser ablation., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Recent experiments have underlined the potential of δ2H in tree-ring cellulose as a physiological indicator of shifts in autotrophic versus heterotrophic processes (i.e., the use of fresh versus stored non-structural carbohydrates). However, the impact of these processes has not yet been quantified under natural conditions. Defoliator outbreaks disrupt tree functioning and carbon assimilation, stimulating remobilization, therefore providing a unique opportunity to improve our understanding of changes in δ2H. By exploring a 700-year tree-ring isotope chronology from Switzerland, we assessed the impact of 79 larch budmoth (LBM, Zeiraphera griseana [Hübner]) outbreaks on the growth of its host tree species, Larix decidua [Mill]. The LBM outbreaks significantly altered the tree-ring isotopic signature, creating a 2H-enrichment and an 18O- and 13C-depletion. Changes in tree physiological functioning in outbreak years are shown by the decoupling of δ2H and δ18O (O-H relationship), in contrast to the positive correlation in non-outbreak years. Across the centuries, the O-H relationship in outbreak years was not significantly affected by temperature, indicating that non-climatic physiological processes dominate over climate in determining δ2H. We conclude that the combination of these isotopic parameters can serve as a metric for assessing changes in physiological mechanisms over time., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Recent climate extremes in Mongolia have ignited a renewed interest in understanding past climate variability over centennial and longer time scales across north-central Asia. Tree-ring width records have been extensively studied in Mongolia as proxies for climate reconstruction, however, the climate and environmental signals of tree-ring stable isotopes from this region need to be further explored. Here, we evaluated a 182-year record of tree-ring δ13C and δ18O from Siberian Pine (Pinus sibirica Du Tour) from a xeric site in central Mongolia (Khorgo Lava) to elucidate the environmental factors modulating these parameters. First, we analyzed the climate sensitivity of tree-ring δ13C and δ18O at Khorgo Lava for comparison with ring-width records, which have been instrumental in reconstructing hydroclimate in central Mongolia over two millennia. We also compared stable isotope records of trees with partial cambial dieback ('strip-bark morphology'), a feature of long-lived conifers growing on resource-limited sites, and trees with a full cambium ('whole-bark morphology'), to assess the inferred leaf-level physiological behavior of these trees. We found that interannual variability in tree-ring δ13C and δ18O reflected summer hydroclimatic variability, and captured recent, extreme drought conditions, thereby complementing ring-width records. The tree-ring δ18O records also had a spring temperature signal and thus expanded the window of climate information recorded by these trees. Over longer time scales, strip-bark trees had an increasing trend in ring-widths, δ13C (and intrinsic water-use efficiency, iWUE) and δ18O, relative to whole-bark trees. Our results suggest that increases in iWUE at this site might be related to a combination of leaf-level physiological responses to increasing atmospheric CO2, recent drought, and stem morphological changes. Our study underscores the potential of stable isotopes for broadening our understanding of past climate in north-central Asia. However, further studies are needed to understand how stem morphological changes might impact stable isotopic trends., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Tree-ring intra-annual stable isotopes (δ13C and δ18O) are powerful tools for revealing plant ecophysiological responses to climatic extremes. We analyzed interannual and fine-scale intra-annual variability of tree-ring δ13C and δ18O in Chinese red pine (Pinus massoniana) from southeastern China to explore environmental drivers and potential trade-offs between the main physiological controls. We show that wet season relative humidity (May-October RH) drove interannual variability of δ18O and intra-annual variability of tree-ring δ18O. It also drove intra-annual variability of tree-ring δ13C, whereas interannual variability was mainly controlled by February-May temperature and September-October RH. Furthermore, intra-annual tree-ring δ18O variability was larger during wet years compared with dry years, whereas δ13C variability was lower during wet years compared with dry years. As a result of these differences in intra-annual variability amplitude, process-based models (we used the Roden model for δ18O and the Farquhar model for δ13C) captured the intra-annual δ18O pattern better in wet years compared with dry years, whereas intra-annual δ13C pattern was better simulated in dry years compared with wet years. This result suggests a potential asymmetric bias in process-based models in capturing the interplay of the different mechanistic processes (i.e., isotopic source and leaf-level enrichment) operating in dry versus wet years. We therefore propose an intra-annual conceptual model considering a dynamic trade-off between the isotopic source and leaf-level enrichment in different tree-ring parts to understand how climate and ecophysiological processes drive intra-annual tree-ring stable isotopic variability under humid climate conditions., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Picea abies (L.) Karst. and Fagus sylvatica (L.) are important tree species in Europe, and the foreseen increase in temperature and vapour pressure deficit (VPD) could increase the vulnerability of these species. However, their physiological performance under climate change at temperate and productive sites is not yet fully understood, especially in uneven-aged stands. Therefore, we investigated tree-ring width and stable isotope chronologies (δ13C/δ18O) of these two species at 10 sites along a climate gradient in Central Europe. In these uneven-aged stands, we compared the year-to-year variability of dominant and suppressed trees for the last 80 years in relation to the sites' spatial distribution and climate. δ18O and δ13C were generally consistent across sites and species, showing high sensitivity to summer VPD, whereas climate correlations with radial growth varied much more and depended on mean local climate. We found no significant differences between dominant and suppressed trees in the response of stable isotope ratios to climate variability, especially within the annual high-frequency signals. In addition, we observed a strikingly high coherence of the high-frequency δ18O variations across long distances with significant correlations above 1500 km, whereas the spatial agreement of δ13C variations was weaker (~700 km). We applied a dual-isotope approach that is based on known theoretical understanding of isotope fractionations to translate the observed changes into physiological components, mainly photosynthetic assimilation rate and stomatal conductance. When separating the chronologies in two time windows and investigating the shifts in isotopes ratios, a significant enrichment of either or both isotope ratios over the last decades can be observed. These results, translated by the dual-isotope approach, indicate a general climate-driven decrease in stomatal conductance. This improved understanding of the physiological mechanisms controlling the short-term variation of the isotopic signature will help to define the performance of these tree species under future climate., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The Canary Islands, an archipelago east of Morocco's Atlantic coast, present steep altitudinal gradients covering various climatic zones from hot deserts to subalpine Mediterranean, passing through fog-influenced cloud forests. Unlike the majority of the Canarian flora, Pinus canariensis C. Sm. ex DC. in Buch grow along most of these gradients, allowing the study of plant functioning in contrasting ecosystems. Here we assess the water sources (precipitation, fog) of P. canariensis and its physiological behavior in its different natural environments. We analyzed carbon and oxygen isotope ratios of water and organics from atmosphere, soil and different plant organs and tissues (including 10-year annual time series of tree-ring cellulose) of six sites from 480 to 1990 m above sea level on the Canary Island La Palma. We found a decreasing δ18O trend in source water that was overridden by an increasing δ18O trend in needle water, leaf assimilates and tree-ring cellulose with increasing altitude, suggesting site-specific tree physiological responses to relative humidity. Fog-influenced and fog-free sites showed similar δ13C values, suggesting photosynthetic activity to be limited by stomatal closure and irradiance at certain periods. In addition, we observed an 18O-depletion (fog-free and timberline sites) and 13C-depletion (fog-influenced and fog-free sites) in latewood compared with earlywood caused by seasonal differences in: (i) water uptake (i.e., deeper ground water during summer drought, fog water frequency and interception) and (ii) meteorological conditions (stem radial growth and latewood δ18O correlated with winter precipitation). In addition, we found evidence for foliar water uptake and strong isotopic gradients along the pine needle axis in water and assimilates. These gradients are likely the reason for an unexpected underestimation of pine needle water δ18O when applying standard leaf water δ18O models. Our results indicate that soil water availability and air humidity conditions are the main drivers of the physiological behavior of pine along the Canary Island's altitudinal gradients., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Tree growth is generally considered to be temperature limited at upper elevation treelines, yet climate factors controlling tree growth at semiarid treelines are poorly understood. We explored the influence of climate on stem growth and stable isotopes for Polylepis tarapacana Philipi, the world's highest elevation tree species, which is found only in the South American Altiplano. We developed tree-ring width index (RWI), oxygen (δ18O) and carbon (δ13C) chronologies for the last 60 years at four P. tarapacana stands located above 4400 m in elevation, along a 500 km latitude aridity gradient. Total annual precipitation decreased from 300 to 200 mm from the northern to the southern sites. We used RWI as a proxy of wood formation (carbon sink) and isotopic tree-ring signatures as proxies of leaf-level gas exchange processes (carbon source). We found distinct climatic conditions regulating carbon sink processes along the gradient. Current growing-season temperature regulated RWI at northern-wetter sites, while prior growing-season precipitation determined RWI at arid southern sites. This suggests that the relative importance of temperature to precipitation in regulating tree growth is driven by site water availability. By contrast, warm and dry growing seasons resulted in enriched tree-ring δ13C and δ18O at all study sites, suggesting that similar climate conditions control carbon-source processes along the gradient. Site-level δ13C and δ18O chronologies were significantly and positively related at all sites, with the strongest relationships among the southern drier stands. This indicates an overall regulation of intercellular carbon dioxide via stomatal conductance for the entire P. tarapacana network, with greater stomatal control when aridity increases. This manuscript also highlights a coupling (decoupling) between physiological processes at leaf level and wood formation as a function of similarities (differences) in their climatic sensitivity. This study contributes to a better understanding and prediction of the response of high-elevation Polylepis woodlands to rapid climate changes and projected drying in the Altiplano., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

The carbon and oxygen isotopic composition of water and assimilates in plants reveals valuable information on plant responses to climatic conditions. Yet, the carbon and oxygen uptake, incorporation and allocation processes determining isotopic compositions are not fully understood. We carried out a dual-isotope labeling experiment at high humidity with 18O-enriched water (H218O) and 13C-enriched CO2 (13CO2) with attached Scots pine (Pinus sylvestris L.) branches and detached twigs of hemiparasitic mistletoes (Viscum album ssp. austriacum) in a naturally dry coniferous forest, where also a long-term irrigation takes place. After 4 h of label exposure, we sampled previous- and recent-year leaves, twig phloem and twig xylem over 192 h for the analysis of isotope ratios in water and assimilates. For both species, the uptake into leaf water and the incorporation of the 18O-label into leaf assimilates was not influenced by soil moisture, while the 13C-label incorporation into assimilates was significantly higher under irrigation compared with control dry conditions. Species-specific differences in leaf morphology or needle age did not affect 18O-label uptake into leaf water, but the incorporation of both tracers into assimilates was two times lower in mistletoe than in pine. The 18O-label allocation in water from pine needles to twig tissues was two times higher for phloem than for xylem under both soil moisture conditions. In contrast, the allocation of both tracers in pine assimilates were similar and not affected by soil moisture, twig tissue or needle age. Soil moisture effects on 13C-label but not on 18O-label incorporation into assimilates can be explained by the stomatal responses at high humidity, non-stomatal pathways for water and isotope exchange reactions. Our results suggest that non-photosynthetic 18O-incorporation processes may have masked prevalent photosynthetic processes. Thus, isotopic variation in leaf water could also be imprinted on assimilates when photosynthetic assimilation rates are low., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

In many regions, drought is suspected to be a cause of Scots pine decline and mortality, but the underlying physiological mechanisms remain unclear. Because of their relationship to ecohydrological processes, δ18O values in tree rings are potentially useful for deciphering long-term physiological responses and tree adaptation to increasing drought. We therefore analyzed both needle- and stem-level isotope fractionations in mature trees exposed to varying water supply. In a first experiment, we investigated seasonal δ18O variations in soil and needle water of Scots pine in a dry inner Alpine valley in Switzerland, comparing drought-stressed trees with trees that were irrigated for more than 10 years. In a second experiment, we analyzed twentieth-century δ18O variations in tree rings of the same forest, including a group of trees that had recently died. We observed less 18O enrichment in needle water of drought-stressed compared with irrigated trees. We applied different isotope fractionation models to explain these results, including the Péclet and the two-pool correction, which considers the ratio of unenriched xylem water in the needles to total needle water. Based on anatomical measurements, we found this ratio to be unchanged in drought-stressed needles, although they were shorter. The observed lower 18O enrichment in needles of stressed trees was therefore likely caused by increased effective path length for water movement within the leaf lamina. In the tree-ring study, we observed lower δ18O values in tree rings of dead trees compared with survivors during several decades prior to their death. These lower values in declining trees are consistent with the lower needle water 18O enrichment observed for drought-stressed compared with irrigated trees, suggesting that this needle-level signal is reflected in the tree rings, although changes in rooting depth could also play a role. Our study demonstrates that long-term effects of drought are reflected in the tree-ring δ18O values, which helps to provide a better understanding of past tree physiological changes of Scots pine., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Compression wood (CW) is a common tissue present in the trunk, branches and roots of mechanically stressed coniferous trees. Its main role is to increase the mechanical strength and regain the vertical orientation of a leaning stem. Compression wood is thought to influence the climate signal in different tree-ring measures. Hence trees containing CW are mostly excluded from tree-ring studies reconstructing past climate variability. There is a large gap of systematic work testing the potential effect of CW on the strength of the climate signal in different tree-ring parameters, especially stable isotope records. Here we test for the first time the effect of CW contained in montane Norway spruce (Picea abies L. Karst) on both δ13C and δ18O tree-ring cellulose records by analyzing compression and opposite wood radii from several disturbed trees together with samples from undisturbed reference trees. We selected four trees tilted by geomorphic processes that were felled by wind and four undisturbed reference trees in the Tatra Mountains, Poland. We qualitatively classified the strength of CW using wood cell anatomical characteristics (tracheid shape, cell wall thickness and presence of intercellular spaces). Then we developed tree-ring width and δ13C and δ18O chronologies from the CW radii, from the opposite radii of the tilted trees and from the reference radii. We tested the effect of CW on tree-ring cellulose δ13C and δ18O variability and on the climate signal strength. We found only minor differences in the means of δ13C and δ18O of compression (δ13C: -22.81‰, δ18O: 28.29‰), opposite (δ13C: -23.02‰; δ18O: 28.05‰) and reference (δ13C: -22.78‰; δ18O: 27.61‰) radii. The statistical relationships between climate variables, δ13C and δ18O, remained consistent among all chronologies. Our findings suggest that moderately tilted trees containing CW can be used to reconstruct past geomorphic activity and for stable isotope-based dendroclimatology., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Altitudinally separated bristlecone pine populations in the White Mountains (California, USA) exhibit differential climate-growth responses as temperature and tree-water relations change with altitude. These populations provide a natural experiment to explore the ecophysiological adaptations of this unique tree species to the twentieth century climate variability. We developed absolutely dated annual ring-width chronologies, and cellulose stable carbon and oxygen isotope chronologies from bristlecone pine growing at the treeline (~3500 m) and ~200 m below for the period AD 1710-2010. These chronologies were interpreted in terms of ecophysiological adaptations to climate variability with a dual-isotope model and a leaf gas exchange model. Ring widths show positive tree growth anomalies at treeline and consistent slower growth below treeline in relation to the twentieth century warming and associated atmospheric drying until the 1980s. Growth rates of both populations declined during and after the 1980s when growing-season temperature and atmospheric vapour pressure deficit continued to increase. Our model-based interpretations of the cellulose stable isotopes indicate that positive treeline growth anomalies prior to the 1980s were related to increased stomatal conductance and leaf-level transpiration and photosynthesis. Reduced growth since the 1980s occurred with a shift to more conservative leaf gas exchange in both the treeline and below-treeline populations, whereas leaf-level photosynthesis continued to increase in response to rising atmospheric CO2 concentrations. Our results suggest that warming-induced atmospheric drying confounds positive growth responses of apparent temperature-limited bristlecone pine populations at treeline. In addition, the observed ecophysiological responses of attitudinally separated bristlecone pine populations illustrate the sensitivity of conifers to climate change., (© The Author(s) 2019. Published by Oxford University Press.)

Isotopes in tropical trees rings can improve our understanding of tree responses to climate. We assessed how climate and growing conditions affect tree-ring oxygen and carbon isotopes (δ18OTR and δ13CTR) in four Amazon trees. We analysed within-ring isotope variation for two terra firme (non-flooded) and two floodplain trees growing at sites with varying seasonality. We find distinct intra-annual patterns of δ18OTR and δ13CTR driven mostly by seasonal variation in weather and source water δ18O. Seasonal variation in isotopes was lowest for the tree growing under the wettest conditions. Tree ring cellulose isotope models based on existing theory reproduced well observed within-ring variation with possible contributions of both stomatal and mesophyll conductance to variation in δ13CTR. Climate analysis reveal that terra firme δ18OTR signals were related to basin-wide precipitation, indicating a source water δ18O influence, while floodplain trees recorded leaf enrichment effects related to local climate. Thus, intrinsically different processes (source water vs leaf enrichment) affect δ18OTR in the two different species analysed. These differences are likely a result of both species-specific traits and of the contrasting growing conditions in the floodplains and terra firme environments. Simultaneous analysis of δ13CTR and δ18OTR supports this interpretation as it shows strongly similar intra-annual patterns for both isotopes in the floodplain trees arising from a common control by leaf stomatal conductance, while terra firme trees showed less covariation between the two isotopes. Our results are interesting from a plant physiological perspective and have implications for climate reconstructions as trees record intrinsically different processes., (© The Author(s) 2019. Published by Oxford University Press.)

In South-western Morocco, water scarcity and high temperature are the main factors determining species survival. Argania spinosa (L.) Skeels is a tree species, endemic to Morocco, which is suffering from ongoing habitat shrinkage. Argan trees play essential local ecological and economic roles: protecting soils from erosion, shading different types of crops, helping maintain soil fertility and, even more importantly, its seeds are used by the local population for oil production, with valuable nutritional, medicinal and cosmetic purposes. The main objective of this study was to identify the sources of water used by this species and to assess the effect of water availability on the photosynthetic rate and stem water potential in two populations: one growing on the coast and a second one 10 km inland. Stem water potential, photosynthetic rate and xylem water isotopic composition (δ18O) were seasonally monitored during 2 years. Trees from both populations showed a similar strategy in the use of the available water sources, which was strongly dependent on deep soil water throughout the year. Nevertheless, during the wet season or under low precipitation a more complex water uptake pattern was found with a mixture of water sources, including precipitation and soil at different depths. No evidence was found of the use of either groundwater or atmospheric water in this species. Despite the similar water-use strategy, the results indicate that Argania trees from the inland population explored deeper layers than coastal ones as suggested by more depleted δ18O values recorded in the inland trees and better photosynthetic performance, hence suggesting that the coastal population of A. spinosa could be subjected to higher stress.

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., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Stable isotope ratios (δ13C and δ18O) of tree-ring α-cellulose are important tools in paleoclimatology, ecology, plant physiology and genetics. The Multiple Sample Isolation System for Solids (MSISS) was a major advance in the tree-ring α-cellulose extraction methods, offering greater throughput and reduced labor input compared to traditional alternatives. However, the usability of the method for resinous conifer species may be limited by the need to remove extractives from some conifer species in a separate pretreatment step. Here we test the necessity of pretreatment for α-cellulose extraction in loblolly pine (Pinus taeda L.), and the efficiency of a modified acetone-based ambient-temperature step for the removal of extractives (i) in loblolly pine from five geographic locations representing its natural range in the southeastern USA, and (ii) on five other common coniferous species (black spruce (Picea mariana Mill.), Fraser fir (Abies fraseri (Pursh) Poir.), Douglas fir (Pseudotsuga menziesii (Mirb.) Franco), Norway spruce (Picea abies (L.) Karst) and ponderosa pine (Pinus ponderosa D.)) with contrasting extractive profiles. The differences of δ13C values between the new and traditional pretreatment methods were within the precision of the isotope ratio mass spectrometry method used (±0.2‰), and the differences between δ18O values were not statistically significant. Although some unanticipated results were observed in Fraser fir, the new ambient-temperature technique was deemed as effective as the more labor-consuming and toxic traditional pretreatment protocol. The proposed technique requires a separate acetone-inert multiport system similar to MSISS, and the execution of both pretreatment and main extraction steps allows for simultaneous treatment of up to several hundred microsamples from resinous softwood, while the need of additional labor input remains minimal.

Understanding how tree growth strategies may influence tree susceptibility to disturbance is an important goal, especially given projected increases in diverse ecological disturbances this century. We use growth responses of tree rings to climate, relationships between tree-ring stable isotopic signatures of carbon (δ(13)C) and oxygen (δ(18)O), wood nitrogen concentration [N], and contemporary leaf [N] and δ(13)C values to assess potential historic drivers of tree photosynthesis in dying and apparently healthy co-occurring northern red oak (Quercus rubra L. (Fagaceae)) during a region-wide oak decline event in Arkansas, USA. Bole growth of both healthy and dying trees responded negatively to drought severity (Palmer Drought Severity Index) and temperature; healthy trees exhibited a positive, but small, response to growing season precipitation. Contrary to expectations, tree-ring δ(13)C did not increase with drought severity. A significantly positive relationship between tree-ring δ(13)C and δ(18)O was evident in dying trees (P < 0.05) but not in healthy trees. Healthy trees' wood exhibited lower [N] than that of dying trees throughout most of their lives (P < 0.05), and we observed a significant, positive relationship (P < 0.05) in healthy trees between contemporary leaf δ(13)C and leaf N (by mass), but not in dying trees. Our work provides evidence that for plants in which strong relationships between δ(13)C and δ(18)O are not evident, δ(13)C may be governed by plant N status. The data further imply that historic photosynthesis in healthy trees was linked to N status and, perhaps, C sink strength to a greater extent than in dying trees, in which tree-ring stable isotopes suggest that historic photosynthesis was governed primarily by stomatal regulation. This, in turn, suggests that assessing the relative dominance of photosynthetic capacity vs stomatal regulation as drivers of trees' C accrual may be a feasible means of predicting tree responses to some disturbance events. Our work demonstrates that a dual isotope, tree-ring approach can be integrated with tree N status to begin to unravel a fundamental question in forest ecology: why do some trees die during a disturbance, while other conspecifics with apparently similar access to resources remain healthy?, (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Most mistletoes are xylem-tapping hemiparasites, which derive their resources from the host's xylem solution. Thus, they affect the host's water relations and resource balance. To understand the physiological mechanisms underlying the mistletoe-host relationship, we experimentally removed Viscum album ssp. austriacum (Wiesb.) Vollmann from adult Pinus sylvestris L. host trees growing in a Swiss dry valley. We analyzed the effects of mistletoe removal over time on host tree growth and on concentrations of nonstructural carbohydrates (NSC) and nitrogen (N) in needles, fine roots and sapwood. In addition, we assessed the δ(13)C and δ(18)O in host tree rings. After mistletoe removal, δ(13)C did not change in newly produced tree rings compared with tree rings in control trees (still infected with mistletoe), but δ(18)O values increased. This pattern might be interpreted as a decrease in assimilation (A) and stomatal conductance (gs), but in our study, it most likely points to an inadequacy of the dual isotope approach. Instead, we interpret the unchanged δ(13)C in tree rings upon mistletoe removal as a balanced increase in A and gs that resulted in a constant intrinsic water use efficiency (defined as A/gs). Needle area-based concentrations of N, soluble sugars and NSC, as well as needle length, single needle area, tree ring width and shoot growth, were significantly higher in trees from which mistletoe was removed than in control trees. This finding suggests that mistletoe removal results in increased N availability and carbon gain, which in turn leads to increased growth rates of the hosts. Hence, in areas where mistletoe is common and the population is large, mistletoe management (e.g., removal) may be needed to improve the host vigor, growth rate and productivity, especially for relatively small trees and crop trees in xeric growth conditions., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

An increase in temperature along with a decrease in summer precipitation in Central Europe will result in an increased frequency of drought events and gradually lead to a change in species composition in forest ecosystems. In the present study, young oaks (Quercus robur L. and Quercus petraea (Matt.) Liebl.) were transplanted into large mesocosms and exposed for 3 years to experimental warming and a drought treatment with yearly increasing intensities. Carbon and oxygen isotopic (δ(13)C and δ(18)O) patterns were analysed in leaf tissue and tree-ring cellulose and linked to leaf physiological measures and tree-ring growth. Warming had no effect on the isotopic patterns in leaves and tree rings, while drought increased δ(18)O and δ(13)C. Under severe drought, an unexpected isotopic pattern, with a decrease in δ(18)O, was observed in tree rings but not in leaves. This decrease in δ(18)O could not be explained by concurrent physiological analyses and is not supported by current physiological knowledge. Analysis of intra-annual tree-ring growth revealed a drought-induced growth cessation that interfered with the record of isotopic signals imprinted on recently formed leaf carbohydrates. This missing record indicates isotopic uncoupling of leaves and tree rings, which may have serious implications for the interpretation of tree-ring isotopes, particularly from trees that experienced growth-limiting stresses., (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Tree-ring stable isotopes, providing insight into drought-induced eco-physiological mechanisms, are frequently used to reconstruct past changes in growing season temperature and precipitation. Their climatic response is, however, still not fully understood, particularly for data originating from non-extreme, mid-latitude environments with differing ecological conditions. Here, we assess the response of δ(13)C, δ(18)O and tree-ring width (TRW) from a temperate mountain forest in the Austrian pre-Alps to climate and specific drought events. Variations in stem growth and isotopic composition of Norway spruce, common beech and European larch from dry, medium and moist sites are compared with records of sunshine, temperature, moisture, precipitation and cloud cover. Results indicate uniform year-to-year variations in δ(13)C and δ(18)O across sites and species, but distinct differences in TRW according to habitat and species. While the climate sensitivity of TRW is overall weak, the δ(13)C and δ(18)O chronologies contain significant signals with a maximum sensitivity to cloud cover changes (r = -0.72 for δ(18)O). The coherent inter-annual isotopic variations are accompanied by substantial differences in the isotopic signatures with offsets up to ∼3‰ for δ(13)C, indicating species-specific physiological strategies and varying water-use efficiencies. During severe summer drought, beech and larch benefit from access to deeper and moist soils, allowing them to keep their stomata open. This strategy is accompanied by an increased water loss through transpiration, but simultaneously enables enhanced photosynthesis. Our findings indicate the potential of tree-ring stable isotopes from temperate forests to reconstruct changes in cloud cover, and to improve knowledge on basic physiological mechanisms of tree species growing in different habitats to cope with soil moisture deficits., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The rising atmospheric CO2 concentration (Ca) has increased tree growth and intrinsic water-use efficiency (iWUE). However, the magnitude of this effect on long-term iWUE and whether this increase could stimulate the growth of riparian forests in extremely arid regions remain poorly understood. We investigated the relationship between growth [ring width; basal area increment (BAI)] and iWUE in a riparian Populus euphratica Oliv. forest to test whether growth was enhanced by increasing CO2 and whether this compensated for environmental stresses in the lower reaches of the inland Heihe River, northwestern China. We accomplished this using dendrochronological methods and carbon (δ(13)C) and oxygen (δ(18)O) isotopic analysis. We found an increase in BAI before 1958, followed by a decrease from 1958 to 1977 and an increase to a peak around 2000. Tree-ring carbon discrimination (Δ) and δ(18)O indicated significant negative overall trends from 1920 to 2012. However, the relationship shifted in strength and direction around 1977 from significantly negative to a weak connection. The seasonal minimum temperature in April to July showed strong influence on Δ, and δ(18)O was controlled by relative humidity (negatively correlated) and temperature (positively correlated) in June and July. The patterns of internal to atmospheric CO2 (Ci/Ca) suggest a specific adaptation of tree physiology to increasing CO2. Intrinsic water-use efficiency increased significantly (by 36.4%) during the study period. The increased iWUE explained 19.8 and 39.1% of the observed yearly and high-frequency (first-order difference) variations in BAI, respectively, after 1977. Our results suggest significant CO2 stimulation of riparian tree growth, which compensated for the negative influences of reductions in river streamflow and a drying climate during the study period., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Dwarf mistletoes, obligate, parasitic plants with diminutive aerial shoots, have long-term effects on host tree water relations, hydraulic architecture and photosynthetic gas exchange and can eventually induce tree death. To investigate the long-term (1886-2010) impacts of dwarf mistletoe on the growth and gas exchange characteristics of host western hemlock, we compared the diameter growth and tree-ring cellulose stable carbon (C) and oxygen (O) isotope ratios (δ(13)Ccell, δ(18)Ocell) of heavily infected and uninfected trees. The relative basal area growth of infected trees was significantly greater than that of uninfected trees in 1886-90, but declined more rapidly in infected than uninfected trees through time and became significantly lower in infected than uninfected trees in 2006-10. Infected trees had significantly lower δ(13)Ccell and δ(18)Ocell than uninfected trees. Differences in δ(18)Ocell between infected and uninfected trees were unexpected given that stomatal conductance and environmental variables that were expected to influence the δ(18)O values of leaf water were similar for both groups. However, estimates of mesophyll conductance (gm) were significantly lower and estimates of effective path length for water movement (L) were significantly higher in leaves of infected trees, consistent with their lower values of δ(18)Ocell. This study reconstructs the long-term physiological responses of western hemlock to dwarf mistletoe infection. The long-term diameter growth and δ(13)Ccell trajectories suggested that infected trees were growing faster than uninfected trees prior to becoming infected and subsequently declined in growth and leaf-level photosynthetic capacity compared with uninfected trees as the dwarf mistletoe infection became severe. This study further points to limitations of the dual-isotope approach for identifying sources of variation in δ(13)Ccell and indicates that changes in leaf internal properties such as gm and L that affect δ(18)Ocell must be considered., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The inter-relationships among δ(13)C and δ(18)O in tree ring cellulose and ring width have the potential to illuminate long-term physiological and environmental information in forest stands that have not been monitored. We examine how within-stand competition and environmental gradients affect ring widths and the stable isotopes of cellulose. We utilize a natural climate gradient across a catchment dominated by Douglas-fir and temporal changes in climate over an 8-year period. We apply a dual-isotope approach to infer physiological response of trees in differing crown dominance classes to temporal and spatial changes in environmental conditions using a qualitative conceptual model of the (13)C-(18)O relationship and by normalizing the data to minimize other variance. The δ(13)C and δ(18)O of cellulose were correlated with year-to-year variation in relative humidity and consistent with current isotope theory. Using a qualitative conceptual model of the (13)C-(18)O relationship and physiological knowledge about the species, we interpreted these changes as stomatal conductance responses to evaporative demand. Spatial variance between plots was not strong and seemed related to leaf nitrogen rather than any other environmental variable. Dominant trees responded to environmental gradients more consistently with current isotope theory as compared with other classes within the same stand. We found a correlation of stable isotopes with environmental variables is useful for assessing the impacts of environmental change over short time series and where growth varies only minimally with climate.

Radial stem growth and the isotopic composition of growth rings are commonly used to quantify the effects of droughts on trees. However, often these parameters are quantified only at one stem height, e.g., 1.3 m, and it is not known how representative this is for the whole stem. This study investigated radial growth at four stem heights (1.3, 5.5, 9.8 and 14 m) of 21, and wood Δ(13)C and δ(18)O at two heights (1.3 and 14 m) of 10 (co-)dominant Norway spruce trees from heavily (HT) and moderately thinned (MT) stands to assess whether different thinning intensities influenced the drought response of stems at different tree heights. Annual basal area increments (BAIs) and stable isotopes in earlywood and latewood were compared between thinning treatments and among the different stem heights. For BAIs, linear correlations with climate were analysed as well. The response of radial growth and isotopic composition to drought was similar at different stem heights in HT trees, but varied with height in MT trees, which were also more sensitive to climatic variations. Recovery of radial growth after drought was more rapid in trees from HT compared with MT stands, except for the topmost height. Basal area increments at breast height (1.3 m) provided good estimates of the volume growth response to drought for the whole stem, but not for its recovery. The faster recovery of radial growth at 1.3 m height of HT compared with MT trees after the 2003 drought was not accompanied by differences in recovery of isotopic composition. However, this is likely to be related to differences between treatments in remobilization of stored C and in tree structure (leaf area, root systems).

A thinning experiment stand at Avoca, Ballinvalley, on the east coast of the Republic of Ireland was used to test a developed methodology aimed at monitoring drought stress, based on the analysis of growth rings obtained by coring. The stand incorporated six plots representing three thinning regimes (light, moderate and heavy) and was planted in the spring of 1943 on a brown earth soil. Radial growth (early- and latewood) was measured for the purpose of this study. A multidisciplinary approach was used to assess historic tree response to climate: specifically, the application of statistical tools such as principal component and canonical correlation analysis to dendrochronology, stable isotopes, ring density proxy, blue reflectance and forest biometrics. Results showed that radial growth was a good proxy for monitoring changes to moisture deficit, while maximum density and blue reflectance were appropriate for assessing changes in accumulated temperature for the growing season. Rainfall also influenced radial growth changes but not significantly, and was a major factor in stable carbon and oxygen discrimination, mostly in the latewood formation phase. Stable oxygen isotope analysis was more accurate than radial growth analysis in drought detection, as it helped detect drought signals in both early- and latewood while radial growth analysis only detected the drought signal in earlywood. Many studies have shown that tree rings provide vital information for marking past climatic events. This work provides a methodology to better identify and understand how commonly measured tree proxies relate to environmental parameters, and can best be used to characterize and pinpoint drought events (variously described using parameters such as like moisture deficit, accumulated temperature, rainfall and potential evaporation).

The carbon and oxygen isotopic composition of tree-ring cellulose was examined in ponderosa pine (Pinus ponderosa Dougl.) trees in the western USA to study seasonal patterns of precipitation inputs. Two sites (California and Oregon) had minimal summer rainfall inputs, whereas a third site (Arizona) received as much as 70% of its annual precipitation during the summer months (North American monsoon). For the Arizona site, both the delta(18)O and delta(13)C values of latewood cellulose increased as the fraction of annual precipitation occurring in the summer (July through September) increased. There were no trends in latewood cellulose delta(18)O with the absolute amount of summer rain at any site. The delta(13)C composition of latewood cellulose declined with increasing total water year precipitation for all sites. Years with below-average total precipitation tended to have a higher proportion of their annual water inputs during the summer months. Relative humidity was negatively correlated with latewood cellulose delta(13)C at all sites. Trees at the Arizona site produced latewood cellulose that was significantly more enriched in (18)O compared with trees at the Oregon or California site, implying a greater reliance on an (18)O-enriched water source. Thus, tree-ring records of cellulose delta(18)O and delta(13)C may provide useful proxy information about seasonal precipitation inputs and the variability and intensity of the North American monsoon.

The Jayme-Wise and diglyme-HCl methods for extracting cellulose from plant material for stable-isotope analysis differ considerably in ease of use, with the latter requiring significantly less time and specialized equipment. However, the diglyme-HCl method leaves a small lignin residue in the crude cellulose that may affect stable-isotope values, whereas alpha-cellulose produced by the Jayme-Wise method is relatively pure. We examined whether adding a bleaching step to the diglyme-HCl method could produce cellulose of comparable purity to alpha-cellulose by comparing the yield, percent carbon, and carbon (delta13C) and oxygen (delta18O) stable isotope ratios of the two celluloses. We tested each method on the wood of five species that differ in ease of delignification, Eucalyptus maculata Hook., E. botryoides Sm., E. resinifera Sm., Pinus pinaster Ait. and Callitris glaucophylla J. Thompson & L.A.S. Johnson, as well as the foliage of C. glaucophylla. For hardwoods such as the eucalypts, the diglyme-HCl method without bleaching produced cellulose with delta13C and delta18O ratios similar to alpha-cellulose. For the softwood, C. glaucophylla, 3 h of bleaching with acidified chlorite following treatment with diglyme-HCl produced cellulose with delta13C and delta18O ratios similar to alpha-cellulose. Bleached and unbleached crude celluloses and alpha-cellulose of P. pinaster were similar in delta18O, but not delta13C. Both types of crude cellulose produced from the foliage of C. glaucophylla had significantly different isotope ratios from alpha-cellulose. Overall, the diglyme-HCl method, with or without bleaching, appears to be a simple, fast method for extracting alpha-cellulose from hardwoods for stable-isotope analyses, but its suitability for softwoods and foliage needs to be evaluated depending on the species.

We measured oxygen isotope ratios (delta18O) of xylem sap, phloem sap, leaves, wood and bark of Eucalyptus globulus Labill. growing in southwestern Australia. Carbon isotope ratios (delta13C) were measured in the dry matter of phloem sap, leaves and wood. Results were used to test several aspects of a mechanistic model of 18O enrichment and provided insights into post-photosynthetic variations in dry matter delta13C. Xylem water delta18O varied little within the tree crown, whereas variation at the landscape-level was more pronounced, with plantations near the coast being enriched by up to 3 per thousand compared with plantations less than 100 km inland. Phloem water was significantly enriched in 18O compared with xylem water in two of three sampling campaigns; mean enrichments were 0.5 and 0.8 per thousand. Phloem sap sugars exported from E. globulus leaves closely reflected observed leaf water enrichment when diurnal variation in photosynthesis was taken into account. Photosynthetic rates were higher in the morning than in the afternoon, whereas leaf water 18O enrichment increased to maximum values in the afternoon. A non-steady-state model of leaf water 18O enrichment accurately predicted observed values through a full diel cycle. Mean estimates of the proportion of organic oxygen effectively exchanging with xylem water during cellulose synthesis were close to 0.40 for both leaves and wood. Carbon isotope ratios of nascent xylem tissues did not differ from those of phloem sap sugars collected concurrently, whereas nascent leaf tissues were depleted in 13C by 2 per thousand compared with phloem sap sugars, suggesting that, in E. globulus, 13C enrichment of sink tissues compared with source leaves does not result from an enriching process within the sink tissue.