Variations in the natural abundance of oxygen-18 and deuterium in plant carbohydrates

Variations in the natural abundance of ¹⁸O and ²H in plant cellulose are influenced by the isotopic composition of the water directly involved in metabolism—the metabolic water fraction. The isotopic distinction between the metabolic source water and total tissue water must reflect the formation of isotopic gradients within the tissue that are influenced by the rate of water turnover, by properties of the water conducting system and by environmental conditions. It seems that the ¹⁸O abundance in the metabolic water is conserved in cellulose with a relatively constant isotope effect. The relationship of the ²H abundance between metabolic water and cellulose is more complex. Hydrogen incorporated into photosynthetic products during primary reduction steps is highly depleted in ²H. However, a large proportion of these hydrogens are subsequently replaced by exchange with water, leading to ²H enrichment during heterotrophic metabolism. Deciphering the oxygen isotope ratio of cellulose could help in providing insights into the carbon and oxygen fluxes exchanged between plants and the atmosphere. This is because the ¹⁸O abundance in cellulose records the ¹⁸O abundance in the metabolic water, which in turn, controls the oxygen isotopic signatures of the CO₂ and O₂ released by plants into the atmosphere. The hydrogen isotope effects associated with carbohydrate metabolism provide insights into the autotrophic state of a plant tissue. This is because the hydrogen isotope ratio of carbohydrates must reflect the net effects of the two opposing isotope effects associated with photosynthesis and heterotrophic metabolism.     

Isotopic heterogeneity of water in transpiring leaves: identification of the component that controls the δ18O of atmospheric O2 and CO2

Two direct but independent approaches were developed to identify the average δ¹⁸O value of the water fraction in the chloroplasts of transpiring leaves. In the first approach, we used the δ¹⁸O value of CO₂ in isotopic equilibrium with leaf water to reconstruct the δ¹⁸O value of water in the chloroplasts. This method was based on the idea that the enzyme carbonic anhydrase facilitates isotopic equilibrium between CO₂ and H₂O predominantly in the chloroplasts, at a rate that is several orders of magnitude faster than the non-catalysed exchange in other leaf water fractions. In the second approach, we measured the δ¹⁸O value of O₂ from photosynthetic water oxidation in the chloroplasts of intact leaves. Since O₂ is produced from chloroplast water irreversibly and without discrimination, the δ¹⁸O value of the O₂ should be identical to that of chloroplast water. In intact, transpiring leaves of sunflower (Helianthus annuus cv. giant mammoth) under the experimental conditions used, the average δ¹⁸O value of chloroplasts water was displaced by 3—10 % (depending on relative humidity and atmospheric composition) below the value predicted by the conventional Craig & Gordon model. Furthermore, this δ¹⁸O value was always lower than the δ¹⁸O value that was measured for bulk leaf water. Our results have implications for a variety of environmental studies since it is the δ¹⁸O value of water in the chloroplasts that is the relevant quantity in considering terrestrial plants influence on the δ¹⁸O values of atmospheric CO₂ and O₂, as well as in influencing the δ¹⁸O of plant organic matter.     

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

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

Natural deuterium and oxygen-18 enrichment in leaf water of cotton plants grown under wet and dry conditions: evidence for water compartmentation and its dynamics

Abstract. Significant differences in leaf water oxygen and hydrogen isotopic composition were observed between cotton plants grown under wet and dry conditions. The magnitude of the differences could be fully explained by the conventional model that describes the isotopic composition of an evaporating water pool under steady state conditions. The results indicate that leaf water isotopic composition is strongly influenced by transpiration rate via its effects on relative humidity adjacent to the leaf surface and on the isotopic composition of the air moisture. Our application of the model, however, provides evidence that leaf water must consist of a mixture of several isotopically distinct pools. These pools are suggested to reside in the symplast, in the cell walls and intercellular spaces and in the veins. A model is proposed suggesting that only the water residing in the cell walls and the intercellular spaces (the transpiration pool) interacts directly with the external environment. The large symplastic pool responds to the external environment to a limited extent via its relatively slow exchange with water in the transpiration pool. It is likely that the isotopic composition of water in the symplastic pool is strongly buffered against shortterm environmental variations, a possibility that would have important implications for the isotopic conditions under which organic matter biosynthesis occurs.     

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

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

植物叶片水稳定同位素研究进展

植物叶片水稳定同位素变化可以直接沟通植物叶片内部与外界的物质和能量联系,并能够反映植物生长周围的气候与生态信息.另外,植物叶片水作为参与水循环的一个重要环节,了解叶片水稳定同位素组成有助于揭示其在局地水体稳定同位素循环中的分配与贡献.概述了国内外叶片水稳定同位素研究现况;介绍了叶片水稳定氢、氧同位素在植物体中的分馏过程及形式(热力学平衡分馏、动力学分馏以及生化分馏)以及影响叶片水稳定同位素组成的气象和生态因子;阐述了叶片水稳定同位素修正的Craig-Gordon稳态模型、string-of-lakes模型、Péclet效应的稳态模型、非稳态效应的模型、Péclet效应的非稳态模型以及二维模型的构建与完善过程;最后讨论了植物叶片水稳定同位素研究存在的问题,并从叶片水稳定同位素与气象、生态因子的关系,叶片水蒸腾线的斜率和截距及过量氘的意义,模型适用性的验证以及叶片水稳定同位素在水文循环的应用等方面展望了研究方向.     

Non-climatic variations in the oxygen isotopic compositions of plants

The ¹⁸O content of leaf water strongly influences the ¹⁸O contents of atmospheric CO₂ and O₂. The ¹⁸O signatures of these atmospheric gases, in turn, emerge as important indicators of large-scale gas exchange processes. Better understanding of the factors that influence the isotopic composition of leaf water is still required, however, for the quantitative utilization of these tracers. The ¹⁸O enrichment of leaf water relative to local meteoric water, is known to reflect climatic conditions. Less is known about the extent variations in the ¹⁸O content of leaf water are influenced by nonclimatic, species-specific characteristics. In a collection of 90 plant species from all continents grown under the same climatic conditions in the Jerusalem Botanical Garden we observed variations of about 9‰ in the δ¹⁸O values of stem water, δ_s, and of about 14‰ in the mid-day δ¹⁸O enrichment of bulk leaf water, δ_LW–δ_s. Differences between δ¹⁸O values predicted by a conventional evaporation model, δ_M, and δ_LW ranged between – 3.3‰ and + 11.8‰. The δ¹⁸O values of water in the chloroplasts (δ_ch) in leaves of 10 selected plants were estimated from on-line CO₂ discrimination measurements. Although much uncertainty is still involved in these estimates, the results indicated that δ_ch can significantly deviate from δ_M in species with high leaf peclet number. The δ¹⁸O values of bulk leaf water significantly correlated with δ¹⁸O values of leaf cellulose (directly) and with instantaneous water use efficiency (A/E, inversely). Differences in isotopic characteristics among conventionally defined vegetation types were not significant, except for conifers that significantly differed from shrubs in δ¹⁸O and δ¹³C values of cellulose and in their peclet numbers, and from deciduous woodland species in their δ¹⁸O and δ¹³C values of cellulose. The results indicated that predictions of the δ¹⁸O values of leaf water (δ_LW, δ_M and δ_ch) could be improved by considering plant species-specific characteristics.     

Transition of stable isotope ratios of leaf water under simulated dew formation

Dew formation, a common meteorological phenomenon, is expected to intensify in the future. Dew can influence the H₂¹⁸O and HDO isotopic compositions of leaf water (δ_L), but the phenomenon has been neglected in many experimental and modelling studies. In this study, the dew effect on δ_L was investigated with a dark plant chamber in which dew formation was introduced. The H₂¹⁸O and HDO compositions of water vapour, dew water and leaf water of five species were measured for up to 48 h of dew exposure. Our results show that the exchanges of H₂¹⁸O and HDO in leaf water with the air continued in the darkness when the net H₂¹⁶O flux was zero. Our estimates of the leaf conductance using the isotopic mass balance method ranged from 0.035 to 0.087 mol m^?2 s^?1, in broad agreement of the night‐time stomatal conductance reported in the literature. In our experiments, the conductance of the C₄ species was 0.04 ± 0.01 mol m^?2 s^?1 and that of the C₃ plants was 0.10 ± 0.04 mol m^?2 s^?1. At the end of 16 h dew exposure, 72 (±17) and 94 (±11)% of the leaf water came from dew according to the ¹⁸O and D tracer, respectively.     

Seasonal variation in δ13C and δ18O of cellulose from growth rings of Pinus radiata

Seasonal variation in δ¹³C and δ¹⁸O of cellulose (δ¹³C_c and δ¹⁸O_c) was measured within two annual rings of Pinus radiata growing at three sites in New Zealand. In general, both δ¹³C_c and δ¹⁸O_c increased to a peak over summer. The three sites differed markedly in annual water balance, and these differences were reflected in δ¹³C_c and δ¹⁸O_c. Average δ¹³C_c and δ¹⁸O_c from each site were positively related, so that the driest site had the most enriched cellulose. δ¹³C_c and δ¹⁸O_c were also related within each site, although both the slope and the closeness of fit of the relationship varied between sites. Supporting the theory, the site with the lowest average relative humidity also had the greatest change in δ¹⁸O_c‰ change in δ¹³C_c. Specific climatic events, such as drought or high rainfall, were recorded as a peak or a trough in enrichment, respectively. These results suggest that seasonal and between‐site variation in δ¹³C_c and δ¹⁸O_c are driven by the interaction between variation in climatic conditions and soil water availability, and plant response to this variation.     

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

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

Life form-specific variations in leaf water oxygen-18 enrichment in Amazonian vegetation

Leaf water (18)O enrichment (Delta(o)) influences the isotopic composition of both gas exchange and organic matter, with Delta(o) values responding to changes in atmospheric parameters. In order to examine possible influences of plant parameters on Delta(o) dynamics, we measured oxygen isotope ratios (delta(18)O) of leaf and stem water on plant species representing different life forms in Amazonia forest and pasture ecosystems. We conducted two field experiments: one in March (wet season) and another in September (dry season) 2004. In each experiment, leaf and stem samples were collected at 2-h intervals at night and hourly during the day for 50 h from eight species including upper-canopy forest trees, upper-canopy forest lianas, and lower-canopy forest trees, a C(4) pasture grass and a C(3) pasture shrub. Significant life form-related differences were detected in (18)O leaf water values. Initial modeling efforts to explain these observations over-predicted nighttime Delta(o) values by as much as 10 per thousand. Across all species, errors associated with measured values of the delta(18)O of atmospheric water vapor (delta(v)) appeared to be largely responsible for the over-predictions of nighttime Delta(o) observations. We could not eliminate collection or storage of water vapor samples as a possible error and therefore developed an alternative, plant-based method for estimating the daily average delta(v) value in the absence of direct (reliable) measurements. This approach differs from the common assumption that isotopic equilibrium exists between water vapor and precipitation water, by including transpiration-based contributions from local vegetation through (18)O measurements of bulk leaf water. Inclusion of both modified delta(v) and non-steady state features resulted in model predictions that more reliably predicted both the magnitude and temporal patterns observed in the data. The influence of life form-specific patterns of Delta(o) was incorporated through changes in the effective path length, an important but little known parameter associated with the Péclet effect.     

脱甲河氢氧同位素组分时空分布特征及其影响因素

河水氢氧稳定同位素特征是研究水体转化和示踪水循环过程的重要内容.为研究河水氢氧稳定同位素特征,揭示河水补给来源,于2017年4—8月对亚热带农业小流域脱甲河4级河段(S_1、S_2、S_3和S_4)水体氢(D)、氧(¹⁸O)稳定同位素进行了监测,分析其时空动态特征和过量氘(d-excess)的变化规律,并探讨了它们与降水、高程和水质等影响因子的相关关系.结果表明:δD、δ¹⁸O和d-excess的变化范围分别在-43.17‰^-26.43‰(-35.50‰±5.44‰)、-7.94‰^-5.70‰(-6.86‰±0.74‰)和16.77‰~23.49‰(19.39‰±1.95‰).受季风环流的影响,δD和δ¹⁸O具有明显的季节变化特征,即春季(δD和δ¹⁸O为-29.88‰±3.31‰和-6.18‰±0.57‰)>夏季(δD和δ¹⁸O为-39.25‰±2.65‰和-7.32‰±0.42‰);空间上,δD和δ¹⁸O表现出明显的沿程变化,随着采样点的位置到河流源头的距离波动增加,δD为S_118O为S_118O与水温呈显著负相关(δD:r=-0.92;δ¹⁸O:r=-0.88);δ¹⁸O与海拔呈显著负相关(r=-0.96);在空间上,δ¹⁸O与水温呈显著正相关(r=0.98);δD和δ¹⁸O与降水量呈不显著负相关.     

Modelling the isotope enrichment of leaf water

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

The influence of transpiration on the equilibration of leaf water with atmospheric water vapour

Abstract. The authors examine the isotopic composition of leaf water, at natural abundance levels, as influenced by transpiration rate. The isotopic composition of water of wheat leaves ( Triticum aestivum L. var. Aroona) was followed while their transpiration rate adjusted to 'steady-state' environmental conditions. Leaf diffusive resistance was modified by short-term salt treatment and by plant culture in either nutrient solution, free-draining sand, or vermiculite. Resultant changes in ¹⁸O and ²H in leaf water are described and fitted to the model of Leaney et al. (1985). The treatments with lower transpiration rates were found to have a greater fraction of their leaf water equilibrated with water vapour in the atmosphere. Comparable results were obtained with both ¹⁸O and ²H, with some differences being interpreted in terms of turbulence in the vapour diffusion path. The fraction of the leaf water equilibrated with the atmosphere varied between leaves of different ages. However, this may have been due to their different positions in the canopy.     

Seasonal patterns in water use and leaf turnover of different plant functional types in a species-rich woodland,south-western Australia

Woodlands in south-western Australia are evergreen and transpire throughout the year despite the long, hot and dry summers of the Mediterranean climate. Results from a case study in a species-rich Banksia woodland are used to discuss the ecological and physiological properties that appear to be essential features of this and similar communities. Tree, shrub and perennial herbaceous species with long-lived leaves dominate the community, whereas winter-green herbaceous species with short-lived leaves constitute a minor group. The total leaf area index is therefore reasonably constant in all seasons. Leaf area index is low and canopies are open, causing good coupling between the vegetation and the atmosphere, and making stomatal control an effective regulator of transpiration. Mean maximum (winter) stomatal conductances were high at approximately 300 mmol m^–2 s^–1. Deep-rootedness allows the dominant species to access soil moisture throughout the unsaturated zone, and down to the capillary fringe of the saturated zone. Shrubs and herbs with shallow roots experience greater drought stress during summer. Rates of community evapotranspiration are limited by leaf area index in the wet season, and further reduced by stomatal closure in the dry season. Deep-rooted plants appear to decrease their stomatal conductance before the development of severe drought stress. Such conservative behaviour, possibly related to plant hydraulic constraints, is a contributing factor to the limited seasonality in community water use.     

Changes in microbial activity and composition in a pasture ecosystem exposed to elevated atmospheric carbon dioxide

Elevated atmospheric CO₂ increases aboveground plant growth and productivity. However, carbon dioxide-induced alterations in plant growth are also likely to affect belowground processes, including the composition of soil biota. We investigated the influence of increased atmospheric CO₂on bacterial numbers and activity, and on soil microbial community composition in a pasture ecosystem under Free-Air Carbon Dioxide Enrichment (FACE). Composition of the soil microbial communities, in rhizosphere and bulk soil, under two atmospheric CO₂ levels was evaluated by using phospholipid fatty acid analysis (PLFA), and total and respiring bacteria counts were determined by epifluorescence microscopy. While populations increased with elevated atmospheric CO₂ in bulk soil of white clover (Trifolium repens L.), a higher atmospheric CO₂ concentration did not affect total or metabolically active bacteria in bulk soil of perennial ryegrass (Lolium perenne L.). There was no effect of atmospheric CO₂ on total bacteria populations per gram of rhizosphere soil. The combined effect of elevated CO₂ on total root length of each species and the bacterial population in these rhizospheres, however, resulted in an 85% increase in total rhizosphere bacteria and a 170% increase in respiring rhizosphere bacteria for the two plant species, when assessed on a per unit land area basis. Differences in microbial community composition between rhizosphere and bulk soil were evident in samples from white clover, and these communities changed in response to CO₂ enrichment. Results of this study indicate that changes in soil microbial activity, numbers, and community composition are likely to occur under elevated atmospheric CO₂, but the extent of those changes depend on plant species and the distance that microbes are from the immediate vicinity of the plant root surface.     

The 18O-signal transfer from water vapour to leaf water and assimilates varies among plant species and growth forms

The ¹⁸O signature of atmospheric water vapour (δ¹⁸O_V) is known to be transferred via leaf water to assimilates. It remains, however, unclear how the ¹⁸O-signal transfer differs among plant species and growth forms. We performed a 9-hr greenhouse fog experiment (relative humidity ≥ 98%) with ¹⁸O-depleted water vapour (−106.7‰) on 140 plant species of eight different growth forms during daytime. We quantified the ¹⁸O-signal transfer by calculating the mean residence time of O in leaf water (MRT_LW) and sugars (MRT_Sugars) and related it to leaf traits and physiological drivers. MRT_LW increased with leaf succulence and thickness, varying between 1.4 and 10.8 hr. MRT_Sugars was shorter in C₃ and C₄ plants than in crassulacean acid metabolism (CAM) plants and highly variable among species and growth forms; MRT_Sugars was shortest for grasses and aquatic plants, intermediate for broadleaf trees, shrubs, and herbs, and longest for conifers, epiphytes, and succulents. Sucrose was more sensitive to δ¹⁸O_V variations than other assimilates. Our comprehensive study shows that plant species and growth forms vary strongly in their sensitivity to δ¹⁸O_V variations, which is important for the interpretation of δ¹⁸O values in plant organic material and compounds and thus for the reconstruction of climatic conditions and plant functional responses.     

Oxygen stable isotope ratios of tree-ring cellulose: the next phase of understanding

Analysis of the oxygen isotope ratio of tree-ring cellulose is a valuable tool that can be used as a paleoclimate proxy. Our ability to use this tool has gone through different phases. The first began in the 1970s with the demonstration of empirical relationships between the oxygen isotope ratio of tree-ring cellulose and climate. These empirical relationships, however, did not provide us with the confidence that they are robust through time, across taxa and across geographical locations. The second phase began with a rudimentary understanding of the physiological and biochemical mechanisms responsible for the oxygen isotope ratios of cellulose, which is necessary to increase the power of this tool. This phase culminated in a mechanistic tree-ring model integrating concepts of physiology and biochemistry in a whole-plant system. This model made several assumptions about leaf water isotopic enrichment and biochemistry which, in the nascent third phase, are now being challenged, with surprising results. These third-phase results suggest that, contrary to the model assumption, leaf temperature across a large latitudinal gradient is remarkably constant and does not follow ambient temperature. Recent findings also indicate that the biochemistry responsible for the incorporation of the cellulose oxygen isotopic signature is not as simple as has been assumed. Interestingly, the results of these challenges have strengthened the tree-ring model. There are several other assumptions that can be investigated which will improve the utility of the tree-ring model.     

Modelling advection and diffusion of water isotopologues in leaves

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