Observed δ13C Values Research Articles

Spatial, Temporal, and Interspecific Differences in Composition of Stable Isotopes in Fishes in Maryland Coastal Bays

Carbon (δ13C) and nitrogen (δ15N) isotopes were used to evaluate spatial, temporal, and interspecific differences in trophic relationships of four fish species (Paralichthys dentatus, Anchoa mitchilli, Leiostomus xanthurus, and Bairdiella chrysoura) in Maryland’s coastal bays. The δ13C values for all species were more enriched in 2017 than in 2018, a year of higher-than-average rainfall that likely caused higher amounts of terrestrial carbon to enter the estuary. There were significant differences among species in the δ13C values, with L. xanthurus being the least depleted (−17.2‰ in 2017; −18.8‰ in 2018). Spatially, the δ13C values of the species, particularly P. dentatus and B. chrysoura, were more depleted in the northern bays, which have a higher nutrient content and receive more freshwater inflow directly from tributaries, than the southern bays. The observed δ13C values (−19.5 ± 0.2‰ to –17.2 ± 0.3‰), however, indicate that marine phytoplankton was the primary carbon source of the fishes. Overall, A. mitchilli was the most enriched in δ15N (13.0‰), and L. xanthurus was the most depleted (10.2‰). δ15N was more enriched in fish from the more human-impacted northern bays than in fish from the southern bays, though this might also have stemmed from the differences in the diet composition of the species in the northern and southern bays. A. mitchilli had the highest trophic level, while L. xanthurus and P. dentatus had the lowest trophic levels. Niche breadth was widest in L. xanthurus compared to the other fish species, suggesting a higher variability in diets among L. xanthurus individuals, leading to specialized diets. There was a high niche overlap between B. chrysoura, A. mitchilli, and L. xanthurus, which indicates they fed on similar prey resources.

Stable Isotopic (δ13C) Evidence for Global Microbial Sequestration of Refractory Dissolved Organic Matter

AbstractDissolved organic carbon (DOC) in the global oceans is an important long‐term carbon sink. Connections between molecular size, reactivity, and isotopic characteristics show that DOC exists on a continuum from biologically reactive to recalcitrant. The driving mechanisms behind the creation and persistence of recalcitrant DOC remain unknown. We show mean recalcitrant DOC (isolated via solid‐phase extraction; SPE‐DOC) δ13C values are 1.3 ± 0.6‰ lower than mean total DOC δ13C between depth ranges 0–200 m and 2–4 km on three GO‐SHIP Repeat Hydrography cruises. Lowest observed δ13C values correlate with low ∆14C and proximity to deep ocean hydrothermal systems. These data support the hypothesis that reworking of DOC through the microbial carbon pump is a key driver of the ocean's long‐term carbon sink. Mass‐balance modeling shows deep‐ocean DOC not captured by SPE is enriched in 13C, highlighting the need for continued research on non‐retained DOC to predict mechanisms that drive ocean carbon storage.

No evidence for carbon enrichment in the mantle source of carbonatites in eastern Africa

Abstract Carbonatites are unusual, carbon-rich magmas thought to form either by the melting of a carbon-rich mantle source or by low-degree partial melting of a carbon-poor (<80 ppm C) mantle followed by protracted differentiation and/or immiscibility. Carbonate-bearing mantle xenoliths from Oldoinyo Lengai (East African Rift), the only active volcano erupting carbonatites, have provided key support for a C-rich mantle source. Here, we report unique microscale O and C isotopic analyses of those carbonates, which are present as interstitial grains in the silicate host lava, veins in the xenoliths, and pseudo-inclusions in olivine xenoliths. The δ18O values vary little, from 19‰ to 29‰, whereas δ13C values are more variable, ranging from –23‰ to +0.5‰. We show that such carbonate δ18O values result from the low-temperature precipitation of carbonate in equilibrium with meteoric water, rather than under mantle conditions. In this framework, the observed δ13C values can be reproduced by Rayleigh distillation driven by carbonate precipitation and associated degassing. Together with petrological evidence of a physical connection between the three types of carbonates, our isotopic data support the pedogenic formation of carbonates in the studied xenoliths by soil-water percolation and protracted crystallization along xenolith cracks. Our results refute a mechanism of C enrichment in the form of mantle carbonates in the mantle beneath the Natron Lake magmatic province and instead support carbonatite formation by low-degree partial melting of a C-poor mantle and subsequent protracted differentiation of alkaline magmas.

13C-depleted moissanites in subduction-related rocks: tracers of slab fluids in the Earth’s mantle and a new diamond exploration tool

A magnesian low-Ti shoshonite dike intruding Archean Norwegian Terrane includes numerous grains of black, blue and gray moissanite (SiC). Moissanite contains inclusions of native Si, Fe and Al, Fe-Cr carbides, Fe-Cr-Mn alloys and diamond. The range of observed δ13C values (-24.8 to -29.6‰) is similar to ophiolite-hosted SiC, lower mantle diamonds and slab-derived biogenic carbon. Norwegian moissanites may have been formed during interaction of Earth’s mantle with carbonaceous slab fluids under extremely reducing conditions as suggested by native metal and carbide inclusions. 13C-depleted moissanite can be used as a new exploration tool for sub-lithospheric diamonds in Archean to Phanerozoic accreted terranes and mobile belts.

Characterization of Microbial Mat Microbiomes in the Modern Thrombolite Ecosystem of Lake Clifton, Western Australia Using Shotgun Metagenomics.

Microbialite-forming communities interact with the environment and influence the precipitation of calcium carbonate through their metabolic activity. The functional genes associated with these metabolic processes and their environmental interactions are therefore critical to microbialite formation. The microbiomes associated with microbialite-forming ecosystems are just now being elucidated and the extent of shared pathways and taxa across different environments is not fully known. In this study, we profiled the microbiome of microbial communities associated with lacustrine thrombolites located in Lake Clifton, Western Australia using metagenomic sequencing and compared it to the non-lithifying mats associated with surrounding sediments to determine whether differences in the mat microbiomes, particularly with respect to metabolic pathways and environmental interactions, may potentially contribute to thrombolite formation. Additionally, we used stable isotope biosignatures to delineate the dominant metabolism associated with calcium carbonate precipitation in the thrombolite build-ups. Results indicated that the microbial community associated with the Lake Clifton thrombolites was predominantly bacterial (98.4%) with Proteobacteria, Cyanobacteria, Bacteroidetes, and Actinobacteria comprising the majority of annotated reads. Thrombolite-associated mats were enriched in photoautotrophic taxa and functional genes associated with photosynthesis. Observed δ13C values of thrombolite CaCO3 were enriched by at least 3.5‰ compared to theoretical values in equilibrium with lake water DIC, which is consistent with the occurrence of photoautotrophic activity in thrombolite-associated microbial mats. In contrast, the microbiomes of microbial communities found on the sandy non-lithifying sediments of Lake Clifton represented distinct microbial communities that varied in taxa and functional capability and were enriched in heterotrophic taxa compared to the thrombolite-associated mats. This study provides new insight into the taxa and functional capabilities that differentiate potentially lithifying mats from other non-lithifying types and suggests that thrombolites are actively accreting and growing in limited areas of Lake Clifton.

Carbonate precipitation in the alkaline lake Specchio di Venere (Pantelleria Island, Italy) and the possible role of microbial mats

Alkaline lakes like the hydrothermally affected lake Specchio di Venere (Pantelleria Island, Central Mediterranean) are typical geological settings harbouring calcified microbial mats. The present work is focused on the discrimination between biotic and abiotic processes driving carbonate precipitation in this lake, using hydrochemical, mineralogical and isotopic data. Hydrochemical analyses demonstrate that the lake is nearly 10−fold supersaturated with regard to aragonite and seasonally reaches hydromagnesite supersaturation. Microscopic observations depict organosedimentary laminated structures consisting of microbial communities and aragonitic precipitates, which are rather disseminated in pores than directly linked to microorganisms. Oxygen isotopic data indicate that authigenic carbonate crystallisation from evaporating water is the dominant precipitation process, further suggested by the absence of textural evidence of diagenetic processes. Conversely, the observed δ13C values reflect an influence of extracellular polymeric substances (EPS) on carbon fractionation during the precipitation process, due to the selective sequestration of 12C in the biomass. The above considerations suggest that at lake Specchio di Venere the carbonate precipitation is mainly of inorganic nature, but a minor role played by biologically influenced processes in microbial mats is not excluded.

Reconstructing C3 and C4 vegetation cover using n-alkane carbon isotope ratios in recent lake sediments from Cameroon, Western Central Africa

Trees and shrubs in tropical Africa use the C3 cycle as a carbon fixation pathway during photosynthesis, while grasses and sedges mostly use the C4 cycle. Leaf-wax lipids from sedimentary archives such as the long-chain n-alkanes (e.g., n-C27 to n-C33) inherit carbon isotope ratios that are representative of the carbon fixation pathway. Therefore, n-alkane δ13C values are often used to reconstruct past C3/C4 composition of vegetation, assuming that the relative proportions of C3 and C4 leaf waxes reflect the relative proportions of C3 and C4 plants. We have compared the δ13C values of n-alkanes from modern C3 and C4 plants with previously published values from recent lake sediments and provide a framework for estimating the fractional contribution (areal-based) of C3 vegetation cover (fC3) represented by these sedimentary archives. Samples were collected in Cameroon, across a latitudinal transect that accommodates a wide range of climate zones and vegetation types, as reflected in the progressive northward replacement of C3-dominated rain forest by C4-dominated savanna. The C3 plants analysed were characterised by substantially higher abundances of n-C29 alkanes and by substantially lower abundances of n-C33 alkanes than the C4 plants. Furthermore, the sedimentary δ13C values of n-C29 and n-C31 alkanes from recent lake sediments in Cameroon (−37.4‰ to −26.5‰) were generally within the range of δ13C values for C3 plants, even when from sites where C4 plants dominated the catchment vegetation. In such cases simple linear mixing models fail to accurately reconstruct the relative proportions of C3 and C4 vegetation cover when using the δ13C values of sedimentary n-alkanes, overestimating the proportion of C3 vegetation, likely as a consequence of the differences in plant wax production, preservation, transport, and/or deposition between C3 and C4 plants. We therefore tested a set of non-linear binary mixing models using δ13C values from both C3 and C4 vegetation as end-members. The non-linear models included a sigmoid function (sine-squared) that describes small variations in the fC3 values as the minimum and maximum δ13C values are approached, and a hyperbolic function that takes into account the differences between C3 and C4 plants discussed above. Model fitting and the estimation of uncertainties were completed using the Monte Carlo algorithm and can be improved by future data addition. Models that provided the best fit with the observed δ13C values of sedimentary n-alkanes were either hyperbolic functions or a combination of hyperbolic and sine-squared functions. Such non-linear models may be used to convert δ13C measurements on sedimentary n-alkanes directly into reconstructions of C3 vegetation cover.

Tracking leakage from a natural CO2 reservoir (Montmiral, France) through the chemistry and isotope signatures of shallow groundwater

AbstractNatural accumulations and releases of CO2 provide the opportunity to study the CO2 trapping and migration mechanisms, the potential impacts of CO2 leaks, and the monitoring tools to assess the impacts of geological storage of anthropogenic CO2. Previous studies on the deep CO2 reservoir of Montmiral (France), focusing on soil gases, groundwater, as well as deep fluids, did not detect any signs of leaks and despite high CO2 fluxes suspiciously high δ13C values have not been stated. In order to further investigate whether some CO2 has leaked from the reservoir toward the surface, we focus here on the major and trace element geochemistry of the shallow aquifers overlying the reservoir with a special focus on the carbonate system, using isotope tracers potentially sensitive to leaks (δ13C of DIC, 87Sr/86Sr and stable isotopes of water). A forward modeling of the potential evolution of groundwater in case of leaks was performed, combining equilibrium calculations of the carbonate system and an ad hoc carbon isotope model. Most observed δ13C values are compatible with modeled carbonate dissolution under open or closed conditions with respect to CO2. A 13C‐enriched subset of samples shows clear signs of incongruent dissolution of Mg‐Sr‐calcite or dolomite, corroborated by 87Sr/86Sr ratios, so that mixing with isotopically heavy deep CO2 is not required to explain the observed chemical and isotope data. The absence of any sign of CO2 leakage into shallow groundwater would support the fact that the reservoir and caprock have been trapping the CO2 efficiently over millions of years.

Timing of isotopic integration in marine mammal skull: comparative study between calcified tissues

Tissues with different turnover rates have different isotope compositions and reflect the different periods in an animal's life when the isotopes are incorporated during the growth of tissues. Bone is one of the most used tissues for reconstruction of an animal's diet; however, the time of isotope integration remains unknown for many species. The δ(15)N and δ(13)C values in tooth dentine and bone tissue from the maxilla and mandible of 21 stranded northern elephant seals, Mirounga angustirostris, collected on the San Benito and Magdalena Islands, Mexico, between 2000 and 2008 were compared. Bone and dentine samples from each growth layer within the tooth were analyzed using a PDZ Europa ANCA-GSL elemental analyzer interfaced with a PDZ Europa 20-20 continuous flow gas source mass spectrometer. The δ(15)N and δ(13)C values were not different between bone structures, indicating similar turnover rates, metabolic activity and amino acid compositions. The differences in the observed δ(13)C values between tissues are probably indicative of differences in their amino acid compositions, although the similarities in δ(15)N values indicated less variation from different amino acids. Correlation of the analyses between isotopic values of tissues suggests that the maxilla and/or mandible of M. angustirostris might reflect the δ(15) N signal incorporated during the last 5 years of life of the individuals. This study demonstrated the usefulness of the applied approach for providing a best approximation of the timing of isotopic integration into the skull of a marine mammal, thereby reducing uncertainty in exploring historic changes in the species' feeding behavior.

Stable isotope and modelling evidence for CO<sub>2</sub> as a driver of glacial–interglacial vegetation shifts in southern Africa

Abstract. Atmospheric CO2 concentration is hypothesized to influence vegetation distribution via tree–grass competition, with higher CO2 concentrations favouring trees. The stable carbon isotope (δ13C) signature of vegetation is influenced by the relative importance of C4 plants (including most tropical grasses) and C3 plants (including nearly all trees), and the degree of stomatal closure – a response to aridity – in C3 plants. Compound-specific δ13C analyses of leaf-wax biomarkers in sediment cores of an offshore South Atlantic transect are used here as a record of vegetation changes in subequatorial Africa. These data suggest a large increase in C3 relative to C4 plant dominance after the Last Glacial Maximum. Using a process-based biogeography model that explicitly simulates 13C discrimination, it is shown that precipitation and temperature changes cannot explain the observed shift in δ13C values. The physiological effect of increasing CO2 concentration is decisive, altering the C3/C4 balance and bringing the simulated and observed δ13C values into line. It is concluded that CO2 concentration itself was a key agent of vegetation change in tropical southern Africa during the last glacial–interglacial transition. Two additional inferences follow. First, long-term variations in terrestrial δ13Cvalues are not simply a proxy for regional rainfall, as has sometimes been assumed. Although precipitation and temperature changes have had major effects on vegetation in many regions of the world during the period between the Last Glacial Maximum and recent times, CO2 effects must also be taken into account, especially when reconstructing changes in climate between glacial and interglacial states. Second, rising CO2 concentration today is likely to be influencing tree–grass competition in a similar way, and thus contributing to the "woody thickening" observed in savannas worldwide. This second inference points to the importance of experiments to determine how vegetation composition in savannas is likely to be influenced by the continuing rise of CO2 concentration.

Quantum mechanical modeling of the structures, energetics and spectral properties of Iα and Iβ cellulose

Periodic planewave and molecular cluster density functional theory (DFT) calculations were performed on Iα and Iβ cellulose in four different conformations each. The results are consistent with the previous interpretation of experimental X-ray and neutron diffraction data that both Iα and Iβ cellulose are dominantly found in the tg conformation of the hydroxymethyl group with a H-bonding conformation termed “Network A”. Structural and energetic results of the periodic DFT calculations with dispersion corrections (DFT-D2) are consistent with observation suggesting that this methodology is accurate to within a few percent for modeling cellulose. The structural and energetic results were confirmed by comparison of calculated vibrational frequencies against observed infrared and Raman frequencies of Iα and Iβ cellulose. Structures extracted from the periodic DFT-D2 energy minimizations were used to calculate the 13C nuclear magnetic resonance chemical shifts (δ13C), and the tg/Network A conformations of both Iα and Iβ cellulose produced excellent correlations with observed δ13C values.

Temperature dependence of the kinetic isotope effect inβ-pinene ozonolysis

[1] The temperature dependence of the kinetic isotope effect (KIE) of β-pinene ozonolysis was investigated experimentally at 258, 273 and 303 K in the AIDA atmospheric simulation chamber. Compound specific carbon isotopic analysis of gas phase samples was performed off-line with a Thermo Desorption-Gas Chromatography-Isotope Ratio Mass Spectrometry (TD-GC-IRMS) system. From the temporal behavior of the δ13C of β-pinene a KIE of 1.00358 ± 0.00013 was derived at 303 K, in agreement with literature data. Furthermore, KIE values of 1.00380 ± 0.00014 at 273 K and 1.00539 ± 0.00012 at 258 K were determined, showing an increasing KIE with decreasing temperature. A parameterization of the observed KIE temperature dependence was deduced and used in a sensitivity study carried out with the global chemistry transport model MOZART-3. Two scenarios were compared, the first neglecting, the second implementing the KIE temperature dependence in the simulations. β-Pinene stable carbon isotope ratio and concentration were computed, with emphasis on boreal zones. For early spring it is shown that when neglecting the temperature dependence of KIE, the calculated average age of β-pinene in the atmosphere can be up to two times over- or underestimated. The evolution of the isotopic composition of the major β-pinene oxidation product, nopinone, was examined using Master Chemical Mechanism (MCM) simulations. The tested hypothesis that formation of nopinone and its associated KIE are the determining factors for the observed δ13C values of nopinone is supported at high β-pinene conversion levels.

Fossil chironomid δ13C as a proxy for past methanogenic contribution to benthic food webs in lakes?

We used a series of experiments to determine whether stable carbon isotope analysis of modern and fossil larval head capsules of chironomids allowed identification of their dietary carbon source. Our main focus was to assess whether carbon from naturally 13C-depleted methane-oxidizing bacteria (MOB) can be traced in chironomid cuticles using stable carbon isotope analysis. We first showed that a minimum sample weight of ~20 μg was required for our equipment to determine head capsule δ13C with a standard deviation of 0.5‰. Such a small minimum sample weight allows taxon-specific δ13C analyses at a precision sufficient to differentiate whether head capsules consist mainly of carbon derived from MOB or from other food sources commonly encountered in lake ecosystems. We then tested the effect of different chemical pre-treatments that are commonly used for sediment processing on δ13C measurements on head capsules. Processing with 10% KOH (2 h), 10% HCl (2 h), or 40% HF (18 h) showed no detectable effect on δ13C, whereas a combination of boiling, accelerated solvent extraction and heavy chemical oxidation resulted in a small (0.2‰) but statistically significant decrease in δ13C values. Using culturing experiments with MOB grown on 13C-labelled methane, we demonstrated that methanogenic carbon is transferred not only into the larval tissue, but also into chironomid head capsules. Taxon-specific δ13C of fossil chironomid head capsules from different lake sediments was analyzed. δ13C of head capsules generally ranged from −28 to −25.8‰, but in some instances we observed δ13C values as low as −36.9 to −31.5‰, suggesting that carbon from MOB is traceable in fossil and subfossil chironomid remains. We demonstrate that stable carbon isotope analyses of fossil chironomid head capsules can give insights into dietary links and carbon cycling in benthic food webs in the past and that the method has the potential to reconstruct the importance of MOB in the palaeo-diet of chironomid larvae and, indirectly, to infer past changes in methane flux at the sediment water interface in lakes.

Reconstruction of past variations of 13C in atmospheric CO2 from its vertical distribution observed in the firn at Dome Fuji, Antarctica

Temporal variations of δ13C of atmospheric CO2 in the past have been reconstructed from the δ13C values of CO2 observed in firn at Dome Fuji, Antarctica. The effective diffusivities of CO2 in firn were estimated for Dome Fuji and another Antarctic site, H72. The age distributions of 13CO2 in firn were first calculated by using a one-dimensional diffusion model, and then the past values of the atmospheric δ13C were derived by using an iterative procedure so that the calculated and observed vertical profiles of δ13C of CO2 in firn agreed with each other. This reconstruction method was also applied to the CH4 concentration to confirm its validity. The values of the atmospheric δ13C thus estimated were in good agreement with those from direct atmospheric measurements at Syowa Station, Antarctica, even for the levelling off of the secular decrease observed in the first half of the 1990s. The statistical uncertainty of the iterative procedure was examined by adding normal pseudo-random numbers to the observed δ13C values in firn. We also calculated the δ13C values for firn at H72 using the reconstructed history of the atmospheric δ13C, and its vertical profile was found to be in close agreement with the observational result.

Unusually low carbon isotope ratios in plants from hanging gardens in southern Utah.

Leaf carbon isotope ratios (δ13C) and photosynthetic gas exchange were measured on plants growing in hanging garden communities in southern Utah, USA. Hanging gardens are unusual, mesic cliff communities occurring where water seeps from the sandstone bedrock in an otherwise extremely arid region; there is very limited overlap in species distributions inside and outside these gardens. Solar exposure in hanging gardens varied with orientation and one of the gardens (Ribbon Garden) was shaded throughout the day. The leaf δ13C values of plants in hanging gardens were significantly more negative than for plants from either nearby ephemeral wash or riparian communities. In Ribbon Garden, the observed δ13C values were as low as -34.8‰, placing them among the most negative values reported for any terrestrial plant species growing in a natural environment. Hanging garden plants were exposed to normal atmospheric CO2 with an average δ13C value of -7.9‰ and so the low leaf δ13C values could not be attributed to exposure to a CO2 source with low 13C content. There was a seasonal change toward more negative leaf δ13C values at the end of the growing season. The observed leaf δ13C values were consistent with photosynthetic gas exchange measurements that indicated unusually high leaf intercellular CO2 concentrations associated with the relatively low light levels in hanging gardens. Thus, extremely negative leaf δ13C values would be expected if significant amounts of the seasonal carbon gain occur at light levels low enough to be near the light compensation point. Maximum observed photosynthetic rates varied with light levels at each of the gardens, with maximum rates averaging 20.3, 14.6, and 3.1 μmol m-2 s-1 at Double Garden, Lost Garden, and Ribbon Garden, respectively. Leaf nitrogen contents averaged 18.5 mg g-1 in species from the more shaded hanging gardens (Lost and Ribbon). When expressed on a leaf area basis, nitrogen contents averaged 117 mmol N m-2 at Lost Garden and 65 mmol N m-2 at Ribbon Garden (shadiest of the two gardens). Leaf nitrogen isotope ratios averaged -2.3‰ (range of -0.7 to -6.1‰), suggesting that most of the nitrogen was derived from a biological fixation source which is most likely the Nostoc growing on the sandstone walls at the seep. These values contrast with leaf nitrogen isotope ratios of 5-9‰ which have been previously reported for arid zone plants in nearby ecosystems.

Depth profiles of δ13C in bottom water and core top C. wuellerstorfi on the Ontong Java Plateau and Emperor Seamounts

We have measured the carbon isotopic composition of dissolved inorganic carbon in bottom waters of the Ontong Java Plateau (western equatorial Pacific) and on the northern Emperor Seamounts (northwest Pacific). Each of these locations is several hundred miles from the nearest Geochemical Ocean Sections Study (GEOSECS) stations, and the observed δ13C values at each site differ substantially from regionally averaged GEOSECS δ13C profiles. We discuss the possible causes of these differences, including horizontal variability, near‐bottom effects, and problems with the Pacific GEOSECS δ13C data. We also measured the isotopic composition (C and O) of core top C. wuellerstorfi from a depth transect of cores at each location. The δ18O data are used to verify that our samples are Holocene. Comparison of foraminiferal and bottom water δ13C values shows that this species faithfully records bottom water δ13C at both sites and demonstrates that there is no depth‐related artifact in the dissolved inorganic carbon‐C. wuellerstorfi δ13C relationship at these sites.

Discrimination between12C and13C by marine plants.

The natural abundance13C/12C ratios (as δ13C) of organic matter of marine macroalgae from Fife and Angus (East Scotland) were measured for comparison with the species' ability to use CO2 and HCO 3- for photosynthesis, as deduced from previously published pH-drift measurements. There was a clear difference in δ13C values for species able or unable to use HCO 3- . Six species of Chlorophyta, 12 species of Phaeophyta and 8 species of Rhodophyta that the pH-drift data suggested could use HCO 3- had δ13C values in the range -8.81‰ to -22.55‰. A further 6 species of Rhodophyta which the pH-drift data suggested could only use CO2 had δ13C values in the range -29.90‰ to-34.51‰. One of these six species (Lomentaria articulata) is intertidal; the other five are subtidal and so have no access to atmospheric CO2 to complicate the analysis. For these species, calculations based on the measured δ13C of the algae, the δ13C of CO2 in seawater, and the known13C/12C discrimination of CO2 diffusion and RUBISCO carboxylation suggest that only 15-21% of the limitation to photosynthesisin situ results from CO2 diffusion from the bulk medium to the plastids; the remaining 79-85% is associated with carboxylation reactions (and, via feedback effects, down-stream processes). This analysis has been extended for one of these five species,Delesseria sanguinea, by incorporating data onin situ specific growth rates, respiratory rates measured in the laboratory, and applying Fick's law of diffusion to calculate a boundary layer thickness of 17-24 μm. This value is reasonable for aDelesseria sanguinea frondin situ. For HCO 3- -using marine macroalgae the range of δ13C values measured can be accommodated by a CO2 efflux from algal cells which range from 0.306 of the gross HCO 3- influx forEnteromorpha intestinalis (δ13C=-8.81‰) in a rockpool to 0.787 forChondrus crispus (δ13C=-22.55‰). The relatively high computed CO2 efflux for those HCO 3- -users with the more negative δ13C values implies a relatively high photon cost of C assimilation; the observed photon costs can be accommodated by assuming coupled, energy-independent inorganic carbon influx and efflux. The observed δ13C values are also interpreted in terms of water movement regimes and obtaining CO2 from the atmosphere. Published δ13C values for freshwater macrophytes were compared with the ability of the species to use CO2 and HCO 3- and again there was an apparent separation in δ13C values for these two groups. δ13C values obtained for marine macroalgae for which no pH-drift data are available permit predictions, as yet untested, as to whether they use predominantly CO2 or HCO 3.

Crassulacean acid metabolism in Kalanchoë species collected in various climatic zones of Madagascar: a survey by δ13C analysis.

The carbon isotope compositions of samples of Kalanchoë species collected at the natural stands in Madagascar were determined. The results suggest that all species of the genus Kalanchoë are capable of crassulacean acid metabolism. The observed δ13C values cover the whole range from -10 to -30‰. This high diversity of the δ13C values was found among the species of the genus as well as, in certain cases, within a single species. This suggest that the CAM patterns in Kalanchoë are generally very flexible. The δ13C values show a clear correlation with the climate of the habitats from where the samples derived. Values indicative of CO2 fixation taking place exclusively during the night were found in the dry regions of Madagascar, whereas δ13C values indicative of mixed CO2 fixation during night and day or of CO2 fixation entirely during the day are distributed in the humid zones.