Nitrogen fertilization and δ18O of CO2 have no effect on 18O‐enrichment of leaf water and cellulose in Cleistogenes squarrosa (C4) – is VPD the sole control?

The oxygen isotope composition of cellulose (δ¹⁸O_Cel) archives hydrological and physiological information. Here, we assess previously unexplored direct and interactive effects of the δ¹⁸O of CO₂ (δ¹⁸O_CO2), nitrogen (N) fertilizer supply and vapour pressure deficit (VPD) on δ¹⁸O_Cel, ¹⁸O‐enrichment of leaf water (Δ¹⁸O_LW) and cellulose (Δ¹⁸O_Cel) relative to source water, and p_exp_x, the proportion of oxygen in cellulose that exchanged with unenriched water at the site of cellulose synthesis, in a C₄ grass (Cleistogenes squarrosa). δ¹⁸O_CO2 and N supply, and their interactions with VPD, had no effect on δ¹⁸O_Cel, Δ¹⁸O_LW, Δ¹⁸O_Cel and p_exp_x. Δ¹⁸O_Cel and Δ¹⁸O_LW increased with VPD, while p_exp_x decreased. That VPD‐effect on p_exp_x was supported by sensitivity tests to variation of Δ¹⁸O_LW and the equilibrium fractionation factor between carbonyl oxygen and water. N supply altered growth and morphological features, but not ¹⁸O relations; conversely, VPD had no effect on growth or morphology, but controlled ¹⁸O relations. The work implies that reconstructions of VPD from Δ¹⁸O_Cel would overestimate amplitudes of VPD variation, at least in this species, if the VPD‐effect on p_exp_x is ignored. Progress in understanding the relationship between Δ¹⁸O_LW and Δ¹⁸O_Cel will require separate investigations of p_ex and p_x and of their responses to environmental conditions.     

Oxygen isotope fractionations across individual leaf carbohydrates in grass and tree species

Almost no δ¹⁸O data are available for leaf carbohydrates, leaving a gap in the understanding of the δ¹⁸O relationship between leaf water and cellulose. We measured δ¹⁸O values of bulk leaf water (δ¹⁸O_LW) and individual leaf carbohydrates (e.g. fructose, glucose and sucrose) in grass and tree species and δ¹⁸O of leaf cellulose in grasses. The grasses were grown under two relative humidity (rH) conditions. Sucrose was generally ¹⁸O‐enriched compared with hexoses across all species with an apparent biosynthetic fractionation factor (ε_bio) of more than 27‰ relative to δ¹⁸O_LW, which might be explained by isotopic leaf water and sucrose synthesis gradients. δ¹⁸O_LW and δ¹⁸O values of carbohydrates and cellulose in grasses were strongly related, indicating that the leaf water signal in carbohydrates was transferred to cellulose (ε_bio = 25.1‰). Interestingly, damping factor p_exp_x, which reflects oxygen isotope exchange with less enriched water during cellulose synthesis, responded to rH conditions if modelled from δ¹⁸O_LW but not if modelled directly from δ¹⁸O of individual carbohydrates. We conclude that δ¹⁸O_LW is not always a good substitute for δ¹⁸O of synthesis water due to isotopic leaf water gradients. Thus, compound‐specific δ¹⁸O analyses of individual carbohydrates are helpful to better constrain (post‐)photosynthetic isotope fractionation processes in plants.     

Interpreting species‐specific variation in tree‐ring oxygen isotope ratios among three temperate forest trees

Although considerable variation has been documented in tree‐ring cellulose oxygen isotope ratios (δ¹⁸O_cell) among co‐occurring species, the underlying causes are unknown. Here, we used a combination of field measurements and modelling to investigate the mechanisms behind variations in late‐wood δ¹⁸O_cell (δ¹⁸O_lc) among three co‐occurring species (chestnut oak, black oak and pitch pine) in a temperate forest. For two growing seasons, we quantified among‐species variation in δ¹⁸O_lc, as well as several variables that could potentially cause the δ¹⁸O_lc variation. Data analysis based on the δ¹⁸O_cell model rules out leaf water enrichment (Δ¹⁸O_lw) and tree‐ring formation period (Δt), but highlights source water δ¹⁸O (δ¹⁸O_sw) as an important driver for the measured difference in δ¹⁸O_lc between black and chestnut oak. However, the enriched δ¹⁸O_lc in pitch pine relative to the oaks could not be sufficiently explained by consideration of the above three variables only, but rather, we show that differences in the proportion of oxygen exchange during cellulose synthesis (p_ex) is most likely a key mechanism. Our demonstration of the relevance of some species‐specific features (or lack thereof) to δ¹⁸O_cell has important implications for isotope based ecophysiological/paleoclimate studies.     

Turnover time of the non‐structural carbohydrate pool influences δ18O of leaf cellulose

Certainty regarding the degree to which organic molecules exchange oxygen with local water during plant cellulose synthesis (p_ex) is necessary for cellulose oxygen isotope (δ¹⁸O_cell)‐based applications in environmental and ecological studies. However, the currently accepted notion that p_ex is a constant of ca. 0.42 appears inconsistent with biochemical theory, which predicts that marked variation may be present in p_ex, in relation to variation in the turnover time (τ) of the carbohydrate pool available for cellulose synthesis. The above prediction was tested in the present study with the analysis of data collected from leaves of Ricinus communis grown in controlled environmental conditions that varied in light intensity and vapour pressure deficit. The results revealed the existence of considerable variation in both p_ex and τ across plants in the various growth environments. Moreover, despite uncertainties in estimates of the proportion of source water in the synthesis water (p_x) and of the biochemical fractionation factor (ε_o), our experiment yielded strong evidence that p_ex exhibits a significant, positive relationship with τ, consistent with biochemical theory. The observed variation in p_ex in association with τ has important implications for the interpretation of δ¹⁸O_cell data in environmental/ecological studies.     

Reconstructing the δ18O of atmospheric water vapour via the CAM epiphyte Tillandsia usneoides: seasonal controls on δ18O in the field and large‐scale reconstruction of δ18Oa

Using both oxygen isotope ratios of leaf water (δ¹⁸O_L) and cellulose (δ¹⁸O_C) of Tillandsia usneoides in situ, this paper examined how short‐ and long‐term responses to environmental variation and model parameterization affected the reconstruction of the atmospheric water vapour (δ¹⁸O_a). During sample‐intensive field campaigns, predictions of δ¹⁸O_L matched observations well using a non‐steady‐state model, but the model required data‐rich parameterization. Predictions from the more easily parameterized maximum enrichment model (δ¹⁸O_L–M) matched observed δ¹⁸O_L and observed δ¹⁸O_a when leaf water turnover was less than 3.5 d. Using the δ¹⁸O_L–M model and weekly samples of δ¹⁸O_L across two growing seasons in Florida, USA, reconstructed δ¹⁸O_a was ?12.6 ± 0.3‰. This is compared with δ¹⁸O_a of ?12.4 ± 0.2‰ resolved from the growing‐season‐weighted δ¹⁸O_C. Both of these values were similar to δ¹⁸O_a in equilibrium with precipitation, ?12.9‰. δ¹⁸O_a was also reconstructed through a large‐scale transect with δ¹⁸O_L and the growing‐season‐integrated δ¹⁸O_C across the southeastern United States. There was considerable large‐scale variation, but there was regional, weather‐induced coherence in δ¹⁸O_a when using δ¹⁸O_L. The reconstruction of δ¹⁸O_a with δ¹⁸O_C generally supported the assumption of δ¹⁸O_a being in equilibrium with precipitation δ¹⁸O (δ¹⁸O_ppt), but the pool of δ¹⁸O_ppt with which δ¹⁸O_a was in equilibrium – growing season versus annual δ¹⁸O_ppt – changed with latitude.     

Leaf δ18O of remaining trees is affected by thinning intensity in a semiarid pine forest

Silvicultural thinning usually improves the water status of remaining trees in water‐limited forests. We evaluated the usefulness of a dual stable isotope approach (δ¹³C, δ¹⁸O) for comparing the physiological performance of remaining trees between forest stands subjected to two different thinning intensities (moderate versus heavy) in a 60‐year‐old Pinus halepensis Mill. plantation in semiarid southeastern Spain. We measured bulk leaf δ¹³C and δ¹⁸O, foliar elemental concentrations, stem water content, stem water δ¹⁸O (δ¹⁸O_{stem water}), tree ring widths and leaf gas exchange rates to assess the influence of forest stand density on tree performance. Remaining trees in low‐density stands (heavily thinned) showed lower leaf δ¹⁸O, and higher stomatal conductance (g_s), photosynthetic rate and radial growth than those in moderate‐density stands (moderately thinned). By contrast, leaf δ¹³C, intrinsic water‐use efficiency, foliar elemental concentrations and δ¹⁸O_{stem water} were unaffected by stand density. Lower foliar δ¹⁸O in heavily thinned stands reflected higher g_s of remaining trees due to decreased inter‐tree competition for water, whereas higher photosynthetic rate was largely attributable to reduced stomatal limitation to CO₂ uptake. The dual isotope approach provided insight into the early (12 months) effects of stand density manipulation on the physiological performance of remaining trees.     

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.     

Genotype differences in 13C discrimination between atmosphere and leaf matter match differences in transpiration efficiency at leaf and whole‐plant levels in hybrid Populus deltoides ×nigra

¹³C discrimination between atmosphere and bulk leaf matter (Δ¹³C_lb) is frequently used as a proxy for transpiration efficiency (TE). Nevertheless, its relevance is challenged due to: (1) potential deviations from the theoretical discrimination model, and (2) complex time integration and upscaling from leaf to whole plant. Six hybrid genotypes of Populus deltoides×nigra genotypes were grown in climate chambers and tested for whole‐plant TE (i.e. accumulated biomass/water transpired). Net CO₂ assimilation rates (A) and stomatal conductance (g_s) were recorded in parallel to: (1) ¹³C in leaf bulk material (δ¹³C_lb) and in soluble sugars (δ¹³C_ss) and (2) ¹⁸O in leaf water and bulk leaf material. Genotypic means of δ¹³C_lb and δ¹³C_ss were tightly correlated. Discrimination between atmosphere and soluble sugars was correlated with daily intrinsic TE at leaf level (daily mean A/g_s), and with whole‐plant TE. Finally, g_s was positively correlated to ¹⁸O enrichment of bulk matter or water of leaves at individual level, but not at genotype level. We conclude that Δ¹³C_lb captures efficiently the genetic variability of whole‐plant TE in poplar. Nevertheless, scaling from leaf level to whole‐plant TE requires to take into account water losses and respiration independent of photosynthesis, which remain poorly documented.     

Contributions of evaporation, isotopic non-steady state transpiration and atmospheric mixing on the delta18O of water vapour in Pacific Northwest coniferous forests

Changes in the ²H and ¹⁸O of atmospheric water vapour provide information for integrating aspects of gas exchange within forest canopies. In this study, we show that diurnal fluctuations in the oxygen isotope ratio (δ¹⁸O) as high as 4‰ were observed for water vapour (δ¹⁸O_vp) above and within an old‐growth coniferous forest in the Pacific Northwest region of the United States. Values of δ¹⁸O_vp decreased in the morning, reached a minimum at midday, and recovered to early‐morning values in the late afternoon, creating a nearly symmetrical diurnal pattern for two consecutive summer days. A mass balance budget was derived and assessed for the ¹⁸O of canopy water vapour over a 2‐d period by considering the ¹⁸O‐isoflux of canopy transpiration, soil evaporation and the air entering the canopy column. The budget was used to address two questions: (1) do δ¹⁸O values of canopy water vapour reflect the biospheric influence, or are such signals swamped by atmospheric mixing? and (2) what mechanisms drive temporal variations of δ¹⁸O_vp? Model calculations show that the entry of air into the canopy column resulted in an isotopically depleted ¹⁸O‐isoflux in the morning of day 1, causing values of δ¹⁸O_vp to decrease. An isotopically enriched ¹⁸O‐isoflux resulting from transpiration then offset this decreased δ¹⁸O_vp later during the day. Contributions of ¹⁸O‐isoflux from soil evaporation were relatively small on day 1 but were more significant on day 2, despite the small H₂¹⁶O fluxes. From measurements of leaf water volume and sapflux, we determined the turnover time of leaf water in the needles of Douglas‐fir trees as ≈ 11 h at midday. Such an extended turnover time suggests that transpiration may not have occurred at the commonly assumed isotopic steady state. We tested a non‐steady state model for predicting δ¹⁸O of leaf water. Our model calculations show that assuming isotopic steady state increased isoflux of transpiration. The impact of this increase on the modelled δ ¹⁸O_vp was clearly detectable, suggesting the importance of considering isotopic non‐steady state of transpiration in studies of forest ¹⁸O water balance.     

Transpiration rate relates to within‐ and across‐species variations in effective path length in a leaf water model of oxygen isotope enrichment

Stable oxygen isotope ratio of leaf water (δ¹⁸O_L) yields valuable information on many aspects of plant–environment interactions. However, current understanding of the mechanistic controls on δ¹⁸O_L does not provide complete characterization of effective path length (L) of the Péclet effect, – a key component of the leaf water model. In this study, we collected diurnal and seasonal series of leaf water enrichment and estimated L in six field‐grown angiosperm and gymnosperm tree species. Our results suggest a pivotal role of leaf transpiration rate (E) in driving both within‐ and across‐species variations in L. Our observation of the common presence of an inverse scaling of L with E in the different species therefore cautions against (1) the conventional treatment of L as a species‐specific constant in leaf water or cellulose isotope (δ¹⁸O_p) modelling; and (2) the use of δ¹⁸O_p as a proxy for g_s or E under low E conditions. Further, we show that incorporation of a multi‐species L‐E scaling into the leaf water model has the potential to both improve the prediction accuracy and simplify parameterization of the model when compared with the conventional approach. This has important implications for future modelling of oxygen isotope ratios.     

Significant Difference in Hydrogen Isotope Composition Between Xylem and Tissue Water in Populus Euphratica

Deuterium depletions between stem water and source water have been observed in coastal halophyte plants and in multiple species under greenhouse conditions. However, the location(s) of the isotope fractionation is not clear yet and it is uncertain whether deuterium fractionation appears in other natural environments. In this study, through two extensive field campaigns utilizing a common dryland riparian tree species Populus euphratica Oliv., we showed that no significant δ¹⁸O differences were found between water source and various plant components, in accord with previous studies. We also found that no deuterium fractionation occurred during P. euphratica water uptake by comparing the deuterium composition (δD) of groundwater and xylem sap. However, remarkable δD differences (up to 26.4‰) between xylem sap and twig water, root water and core water provided direct evidence that deuterium fractionation occurred between xylem sap and root or stem tissue water. This study indicates that deuterium fractionation could be a common phenomenon in drylands, which has important implications in plant water source identification, palaeoclimate reconstruction based on wood cellulose and evapotranspiration partitioning using δD of stem water.     

Dentine oxygen isotopes (δ18O) as a proxy for odontocete distributions and movements

Spatial variation in marine oxygen isotope ratios (δ¹⁸O) resulting from differential evaporation rates and precipitation inputs is potentially useful for characterizing marine mammal distributions and tracking movements across δ¹⁸O gradients. Dentine hydroxyapatite contains carbonate and phosphate that precipitate in oxygen isotopic equilibrium with body water, which in odontocetes closely tracks the isotopic composition of ambient water. To test whether dentine oxygen isotope composition reliably records that of ambient water and can therefore serve as a proxy for odontocete distribution and movement patterns, we measured δ¹⁸O values of dentine structural carbonate (δ¹⁸O_SC) and phosphate (δ¹⁸O_P) of seven odontocete species (n = 55 individuals) from regional marine water bodies spanning a surface water δ¹⁸O range of several per mil. Mean dentine δ¹⁸O_SC (range +21.2 to +25.5‰ VSMOW) and δ¹⁸O_P (+16.7 to +20.3‰) values were strongly correlated with marine surface water δ¹⁸O values, with lower dentine δ¹⁸O_SC and δ¹⁸O_P values in high‐latitude regions (Arctic and Eastern North Pacific) and higher values in the Gulf of California, Gulf of Mexico, and Mediterranean Sea. Correlations between dentine δ¹⁸O_SC and δ¹⁸O_P values with marine surface water δ¹⁸O values indicate that sequential δ¹⁸O measurements along dentine, which grows incrementally and archives intra‐ and interannual isotopic composition over the lifetime of the animal, would be useful for characterizing residency within and movements among water bodies with strong δ¹⁸O gradients, particularly between polar and lower latitudes, or between oceans and marginal basins.     

An oxygen isotope chronometer for cellulose deposition: the successive leaves formed by tillers of a C4 perennial grass

Multiannual time series of (palaeo)hydrological information can be reconstructed from the oxygen isotope composition of cellulose (δ¹⁸O_Cel) in biological archives, for example, tree rings, but our ability to temporally resolve information at subannual scale is limited. We capitalized on the short and predictable leaf appearance interval (2.4 d) of a perennial C₄ grass (Cleistogenes squarrosa), to assess its potential for providing highly time‐resolved δ¹⁸O_Cel records of vapour pressure deficit (VPD). Plants grown at low (0.63 kPa) or high (1.58 kPa) VPD were swapped between VPD environments and exposed to the new environment for 7 d with simultaneous ¹³CO₂ labelling. Then, leaves were sampled by age/position along individual tillers. Five leaves at different developmental stages were growing simultaneously. The period of most‐active leaf elongation, from 10 to 90% of final length, lasted 6.6 d, and ~80% of all carbon and oxygen incorporation in whole‐leaf cellulose occurred within 7 d. Cellulose deposition stopped at (or shortly after) full leaf expansion. The direction of change, low‐to‐high or high‐to‐low VPD, had no differential effect on new oxygen and carbon incorporation in cellulose. Successive leaves produced by tillers of C. squarrosa provide a δ¹⁸O_Cel record useful for reconstructions of short‐term hydrological dynamics.     

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.     

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.     

Carbon and nitrogen isotope fractionation of amino acids in an avian marine predator,the gentoo penguin (Pygoscelis papua)

Compound‐specific stable isotope analysis (CSIA) of amino acids (AA) has rapidly become a powerful tool in studies of food web architecture, resource use, and biogeochemical cycling. However, applications to avian ecology have been limited because no controlled studies have examined the patterns in AA isotope fractionation in birds. We conducted a controlled CSIA feeding experiment on an avian species, the gentoo penguin (Pygoscelis papua), to examine patterns in individual AA carbon and nitrogen stable isotope fractionation between diet (D) and consumer (C) (Δ¹³C_C‐D and Δ¹⁵N_C‐D, respectively). We found that essential AA δ¹³C values and source AA δ¹⁵N values in feathers showed minimal trophic fractionation between diet and consumer, providing independent but complimentary archival proxies for primary producers and nitrogen sources respectively, at the base of food webs supporting penguins. Variations in nonessential AA Δ¹³C_C‐D values reflected differences in macromolecule sources used for biosynthesis (e.g., protein vs. lipids) and provided a metric to assess resource utilization. The avian‐specific nitrogen trophic discrimination factor (TDF_Glu‐Phe = 3.5 ± 0.4‰) that we calculated from the difference in trophic fractionation (Δ¹⁵N_C‐D) of glutamic acid and phenylalanine was significantly lower than the conventional literature value of 7.6‰. Trophic positions of five species of wild penguins calculated using a multi‐TDF_Glu‐Phe equation with the avian‐specific TDF_Glu‐Phe value from our experiment provided estimates that were more ecologically realistic than estimates using a single TDF_Glu‐Phe of 7.6‰ from the previous literature. Our results provide a quantitative, mechanistic framework for the use of CSIA in nonlethal, archival feathers to study the movement and foraging ecology of avian consumers.     

Stable isotope fractionation of strontium in coccolithophore calcite: Influence of temperature and carbonate chemistry

Marine calcifying eukaryotic phytoplankton (coccolithophores) is a major contributor to the pelagic production of CaCO₃ and plays an important role in the biogeochemical cycles of C, Ca and other divalent cations present in the crystal structure of calcite. The geochemical signature of coccolithophore calcite is used as palaeoproxy to reconstruct past environmental conditions and to understand the underlying physiological mechanisms (vital effects) and precipitation kinetics. Here, we present the stable Sr isotope fractionation between seawater and calcite (Δ^88/86Sr) of laboratory cultured coccolithophores in individual dependence of temperature and seawater carbonate chemistry. Coccolithophores were cultured within a temperature and a pCO₂ range from 10 to 25°C and from 175 to 1,240 μatm, respectively. Both environmental drivers induced a significant linear increase in coccolith stable Sr isotope fractionation. The temperature correlation at constant pCO₂ for Emiliania huxleyi and Coccolithus braarudii is expressed as Δ^88/86Sr = ?7.611 × 10^?3 T + 0.0061. The relation of Δ^88/86Sr to pCO₂ was tested in Emiliania huxleyi at 10 and 20°C and resulted in Δ^88/86Sr = ?5.394 × 10^?5 pCO₂ – 0.0920 and Δ^88/86Sr = ?5.742 × 10^?5 pCO₂ – 0.1351, respectively. No consistent relationship was found between coccolith Δ^88/86Sr and cellular physiology impeding a direct application of fossil coccolith Δ^88/86Sr as coccolithophore productivity proxy. An overall significant correlation was detected between the elemental distribution coefficient (D_Sr) and Δ^88/86Sr similar to inorganic calcite with a physiologically induced offset. Our observations indicate (i) that temperature and pCO₂ induce specific effects on coccolith Δ^88/86Sr values and (ii) that strontium elemental ratios and stable isotope fractionation are mainly controlled by precipitation kinetics when embedded into the crystal lattice and subject to vital effects during the transmembrane transport from seawater to the site of calcification. These results provide an important step to develop a coccolith Δ^88/86Sr palaeoproxy complementing the existing toolbox of palaeoceanography.     

The intramolecular ¹³C-distribution in ethanol reveals the influence of the CO₂ -fixation pathway and environmental conditions on the site-specific ¹³C variation in glucose

Efforts to understand the cause of ¹²C versus ¹³C isotope fractionation in plants during photosynthesis and post‐photosynthetic metabolism are frustrated by the lack of data on the intramolecular ¹³C‐distribution in metabolites and its variation with environmental conditions. We have exploited isotopic carbon‐13 nuclear magnetic resonance (¹³C NMR) spectrometry to measure the positional isotope composition (δ¹³C_i, ‰) in ethanol samples from different origins: European wines, liquors and sugars from C₃, C₄ and crassulacean acid metabolism (CAM) plants. In C₃‐ethanol samples, the methylene group was always ¹³C‐enriched (～2‰) relative to the methyl group. In wines, this pattern was correlated with both air temperature and δ¹⁸O of wine water, indicating that water vapour deficit may be a critical defining factor. Furthermore, in C₄‐ethanol, the reverse relationship was observed (methylene‐C relatively ¹³C‐depleted), supporting the concept that photorespiration is the key metabolic process leading to the ¹³C distribution in C₃‐ethanol. By contrast, in CAM‐ethanol, the isotopic pattern was similar to but stronger than C₃‐ethanol, with a relative ¹³C‐enrichment in the methylene‐C of up to 13‰. Plausible causes of this ¹³C‐pattern are briefly discussed. As the intramolecular δ¹³C_i‐values in ethanol reflect that in source glucose, our data point out the crucial impact on the ratio of metabolic pathways sustaining glucose synthesis.     

The influence of (photo)respiration on carbon isotope discrimination in plants

The contribution which (photo)respiration makes to carbon isotope discrimination (Δ¹³C) was examined by conducting simultaneous gas exchange measurements and isotopic analysis of carbon dioxide passing over leaves of Triticum aestivum and Phaseolus vulgaris, via manipulations of the carbon isotope composition (δ¹³C) of source CO₂ during growth and measurement. Dark respiration only altered net Δ¹³C (Δ_obs) at low CO₂ assimilation, and was sensitive to source CO₂δ¹³C during measurement. Photorespiration reduced Δ_obs relative to Δ¹³C predicted from p_i/p_a (Δ_i) over the full range of CO₂ assimilation, to a greater degree under elevated oxygen partial pressure (pO₂), indicating fractionation during photorespiration (f) in T. aestivum. For P. vulgaris, Δ_obs was insensitive to elevated pO₂ at higher assimilation rates, suggesting that f was minimal. A model was developed to calculate gross discrimination (Δ_ps), independent of (photo)respiration, from which estimates of f were obtained for T. aestivum (3.3‰) and P. vulgaris (0.5‰). Because photorespiratory fractionation varies interspecifically, and influences net Δ¹³C which is directly reflected in leaf δ¹³C, consideration of (photo)respiratory fractionation is necessary when interpreting δ¹³C of leaf material, especially under conditions where (photo)respiratory CO₂ losses make a large relative contribution to total plant carbon budgets.     

Dual Δ¹³C/δ¹⁸O response to water and nitrogen availability and its relationship with yield in field-grown durum wheat

The combined use of stable carbon and oxygen isotopes in plant matter is a tool of growing interest in cereal crop management and breeding, owing to its relevance for assessing the photosynthetic and transpirative performance under different growing conditions including water and N regimes. However, this method has not been applied to wheat grown under real field conditions. Here, plant growth, grain yield (GY) and the associated agronomic components, carbon isotope discrimination (Δ¹³C) plus oxygen isotope composition (δ¹⁸O) as well as leaf and canopy gas exchange were measured in field‐grown wheat subjected to different water and N availabilities. Water limitation was the main factor affecting yield, leaf and canopy gas exchange and Δ¹³C and δ¹⁸O, whereas N had a smaller effect on such traits. The combination of Δ¹³C and δ¹⁸O gave a clear advantage compared with gas exchange measurements, as it provides information on the instantaneous and the long‐term plant photosynthetic and transpirative performance and are less labour intensive than gas exchange measurements. In addition, the combination of plant Δ¹³C and δ¹⁸O predicted differences in GY and related agronomical parameters, providing agronomists and breeders with integrative traits for selecting crop management practices and/or genotypes with better performance under water‐limiting and N‐limiting conditions.