Using Carbon Stable Isotopes to Study C3 and C4 Photosynthesis: Models and Calculations.

Stable carbon isotopes are a powerful tool to study photosynthesis. Initial applications consisted of determining isotope ratios of plant biomass using mass spectrometry. Subsequently, theoretical models relating C isotope values to gas exchange characteristics were introduced and tested against instantaneous online measurements of 13C photosynthetic discrimination. Beginning in the twenty-first century, laser absorption spectroscopes with sufficient precision for determining isotope mixing ratios became commercially available. This has allowed collection of large data sets at lower cost and with unprecedented temporal resolution. More data and accompanying knowledge have permitted refinement of 13C discrimination model equations, but often at the expense of increased model complexity and difficult parametrization. This chapter describes instantaneous online measurements of 13C photosynthetic discrimination, provides recommendations for experimental setup, and presents a thorough compilation of equations available to researchers. We update our previous 2018 version of this chapter by including recently improved descriptions of (photo)respiratory processes and associated fractionations. We discuss the capabilities and limitations of the diverse 13C discrimination model equations and provide guidance for selecting the model complexity needed for different applications.

Historical tree phenology data reveal the seasonal rhythms of the Congo Basin rainforest.

Tropical forest phenology directly affects regional carbon cycles, but the relation between species-specific and whole-canopy phenology remains largely uncharacterized. We present a unique analysis of historical tropical tree phenology collected in the central Congo Basin, before large-scale impacts of human-induced climate change. Ground-based long-term (1937-1956) phenological observations of 140 tropical tree species are recovered, species-specific phenological patterns analyzed and related to historical meteorological records, and scaled to characterize stand-level canopy dynamics. High phenological variability within and across species and in climate-phenology relationships is observed. The onset of leaf phenophases in deciduous species was triggered by drought and light availability for a subset of species and showed a species-specific decoupling in time along a bi-modal seasonality. The majority of the species remain evergreen, although central African forests experience relatively low rainfall. Annually a maximum of 1.5% of the canopy is in leaf senescence or leaf turnover, with overall phenological variability dominated by a few deciduous species, while substantial variability is attributed to asynchronous events of large and/or abundant trees. Our results underscore the importance of accounting for constituent signals in canopy-wide scaling and the interpretation of remotely sensed phenology signals.

Contrasting stem water uptake and storage dynamics of water-saver and water-spender species during drought and recovery.

Drought is projected to occur more frequently and intensely in the coming decades, and the extent to which it will affect forest functioning will depend on species-specific responses to water stress. Aiming to understand the hydraulic traits and water dynamics behind water-saver and water-spender strategies in response to drought and recovery, we conducted a pot experiment with two species with contrasting physiological strategies, Scots pine (Pinus sylvestris L.) and Portuguese oak (Quercus faginea L.). We applied two cycles of soil drying and recovery and irrigated with isotopically different water to track fast changes in soil and stem water pools, while continuously measuring physiological status and xylem water content from twigs. Our results provide evidence for a tight link between the leaf-level response and the water uptake and storage patterns in the stem. The water-saver strategy of pines prevented stem dehydration by rapidly closing stomata which limited their water uptake during the early stages of drought and recovery. Conversely, oaks showed a less conservative strategy, maintaining transpiration and physiological activity under dry soil conditions, and consequently becoming more dehydrated at the stem level. We interpreted this dehydration as the release of water from elastic storage tissues as no major loss of hydraulic conductance occurred for this species. After soil rewetting, pines recovered pre-drought leaf water potential rapidly, but it took longer to replace the water from conductive tissues (slower labeling speed). In contrast, water-spender oaks were able to quickly replace xylem water during recovery (fast labeling speed), but it took longer to refill stem storage tissues, and hence to recover pre-drought leaf water potential. These different patterns in sap flow rates, speed and duration of the labeling reflected a combination of water-use and storage traits, linked to the leaf-level strategies in response to drought and recovery.

Water taken up through the bark is detected in the transpiration stream in intact upper-canopy branches.

Alternative water uptake pathways through leaves and bark complement water supply with interception, fog or dew. Bark water-uptake contributes to embolism-repair, as demonstrated in cut branches. We tested whether bark water-uptake could also contribute to supplement xylem-water for transpiration. We applied bandages injected with 2 H-enriched water on intact upper-canopy branches of Pinus sylvestris and Fagus sylvatica in a boreal and in a temperate forest, in summer and winter, and monitored transpiration and online isotopic composition (δ2 H and δ18 O) of water vapour, before sampling for analyses of δ2 H and δ18 O in tissue waters. Xylem, bark and leaf waters from segments downstream from the bandages were 2 H-enriched whereas δ18 O was similar to controls. Transpiration was positively correlated with 2 H-enrichment. Isotopic compositions of transpiration and xylem water allowed us to calculate isotopic exchange through the bark via vapour exchange, which was negligible in comparison to estimated bark water-uptake, suggesting that water-uptake occurred via liquid phase. Results were consistent across species, forests and seasons, indicating that bark water-uptake may be more ubiquitous than previously considered. We suggest that water taken up through the bark could be incorporated into the transpiration stream, which could imply that sap-flow measurements underestimate transpiration when bark is wet.

Do 2 H and 18 O in leaf water reflect environmental drivers differently?

We compiled hydrogen and oxygen stable isotope compositions (δ2 H and δ18 O) of leaf water from multiple biomes to examine variations with environmental drivers. Leaf water δ2 H was more closely correlated with δ2 H of xylem water or atmospheric vapour, whereas leaf water δ18 O was more closely correlated with air relative humidity. This resulted from the larger proportional range for δ2 H of meteoric waters relative to the extent of leaf water evaporative enrichment compared with δ18 O. We next expressed leaf water as isotopic enrichment above xylem water (Δ2 H and Δ18 O) to remove the impact of xylem water isotopic variation. For Δ2 H, leaf water still correlated with atmospheric vapour, whereas Δ18 O showed no such correlation. This was explained by covariance between air relative humidity and the Δ18 O of atmospheric vapour. This is consistent with a previously observed diurnal correlation between air relative humidity and the deuterium excess of atmospheric vapour across a range of ecosystems. We conclude that 2 H and 18 O in leaf water do indeed reflect the balance of environmental drivers differently; our results have implications for understanding isotopic effects associated with water cycling in terrestrial ecosystems and for inferring environmental change from isotopic biomarkers that act as proxies for leaf water.

Evidence for distinct isotopic compositions of sap and tissue water in tree stems: consequences for plant water source identification.

The long-standing hypothesis that the isotopic composition of plant stem water reflects that of source water is being challenged by studies reporting bulk water from woody stems with an isotopic composition that cannot be attributed to any potential water source. The mechanism behind such source-stem water isotopic offsets is still poorly understood. Using a novel technique to extract selectively sap water from xylem conduits, we show that, in cut stems and potted plants, the isotopic composition of sap water reflects that of irrigation water, demonstrating unambiguously that no isotopic fractionation occurs during root water uptake or sap water extraction. By contrast, water in nonconductive xylem tissues is always depleted in deuterium compared with sap water, irrespective of wood anatomy. Previous studies have shown that isotopic heterogeneity also exists in soils at the pore scale in which water adsorbed onto soil particles is more depleted in deuterium than unbound water. Data collected at a riparian forest indicated that sap water matches best unbound soil water from depth below -70 cm, while bulk stem and soil water differ markedly. We conclude that source-stem isotopic offsets can be explained by micrometre-scale heterogeneity in the isotope ratios of water within woody stems and soil micro-pores.

Temperature-sensitive biochemical 18 O-fractionation and humidity-dependent attenuation factor are needed to predict δ18 O of cellulose from leaf water in a grassland ecosystem.

We explore here our mechanistic understanding of the environmental and physiological processes that determine the oxygen isotope composition of leaf cellulose (δ18 Ocellulose ) in a drought-prone, temperate grassland ecosystem. A new allocation-and-growth model was designed and added to an 18 O-enabled soil-vegetation-atmosphere transfer model (MuSICA) to predict seasonal (April-October) and multi-annual (2007-2012) variation of δ18 Ocellulose and 18 O-enrichment of leaf cellulose (Δ18 Ocellulose ) based on the Barbour-Farquhar model. Modelled δ18 Ocellulose agreed best with observations when integrated over c. 400 growing-degree-days, similar to the average leaf lifespan observed at the site. Over the integration time, air temperature ranged from 7 to 22°C and midday relative humidity from 47 to 73%. Model agreement with observations of δ18 Ocellulose (R2  = 0.57) and Δ18 Ocellulose (R2  = 0.74), and their negative relationship with canopy conductance, was improved significantly when both the biochemical 18 O-fractionation between water and substrate for cellulose synthesis (εbio , range 26-30‰) was temperature-sensitive, as previously reported for aquatic plants and heterotrophically grown wheat seedlings, and the proportion of oxygen in cellulose reflecting leaf water 18 O-enrichment (1 - pex px , range 0.23-0.63) was dependent on air relative humidity, as observed in independent controlled experiments with grasses. Understanding physiological information in δ18 Ocellulose requires quantitative knowledge of climatic effects on pex px and εbio .

Asymmetric responses of ecosystem productivity to rainfall anomalies vary inversely with mean annual rainfall over the conterminous United States.

The CONterminous United States (CONUS) presents a large range of climate conditions and biomes where terrestrial primary productivity and its inter-annual variability are controlled regionally by rainfall and/or temperature. Here, the response of ecosystem productivity to those climate variables was investigated across different biomes from 2010 to 2018 using three climate datasets of precipitation, air temperature or drought severity, combined with several proxies of ecosystem productivity: a remote sensing product of aboveground biomass, an net primary productivity (NPP) remote sensing product, an NPP model-based product and four gross primary productivity products. We used an asymmetry index (AI) where positive AI indicates a greater increase of ecosystem productivity in wet years compared to the decline in dry years, and negative AI indicates a greater decline of ecosystem productivity in dry years compared to the increase in wet years. We found consistent spatial patterns of AI across the CONUS for the different products, with negative asymmetries over the Great Plains and positive asymmetries over the southwestern CONUS. Shrubs and, to a lesser extent, evergreen forests show a persistent positive asymmetry, whilst (natural) grasslands appear to have transitioned from positive to negative anomalies during the last decade. The general tendency of dominant negative asymmetry response for ecosystem productivity across the CONUS appears to be influenced by the negative asymmetry of precipitation anomalies. AI was found to be a function of mean rainfall: more positive AIs were found in dry areas where plants are adapted to drought and take advantage of rainfall pulses, and more negative AIs were found in wet areas, with a threshold delineating the two regimes corresponding to a mean annual rainfall of 200-400 mm/year.

µgreen-db: a reference database for the 23S rRNA gene of eukaryotic plastids and cyanobacteria.

Studying the ecology of photosynthetic microeukaryotes and prokaryotic cyanobacterial communities requires molecular tools to complement morphological observations. These tools rely on specific genetic markers and require the development of specialised databases to achieve taxonomic assignment. We set up a reference database, called µgreen-db, for the 23S rRNA gene. The sequences were retrieved from generalist (NCBI, SILVA) or Comparative RNA Web (CRW) databases, in addition to a more original approach involving recursive BLAST searches to obtain the best possible sequence recovery. At present, µgreen-db includes 2,326 23S rRNA sequences belonging to both eukaryotes and prokaryotes encompassing 442 unique genera and 736 species of photosynthetic microeukaryotes, cyanobacteria and non-vascular land plants based on the NCBI and AlgaeBase taxonomy. When PR2/SILVA taxonomy is used instead, µgreen-db contains 2,217 sequences (399 unique genera and 696 unique species). Using µgreen-db, we were able to assign 96% of the sequences of the V domain of the 23S rRNA gene obtained by metabarcoding after amplification from soil DNA at the genus level, highlighting good coverage of the database. µgreen-db is accessible at http://microgreen-23sdatabase.ea.inra.fr.

An explanation for the isotopic offset between soil and stem water in a temperate tree species.

A growing number of field studies report isotopic offsets between stem water and its potential sources that prevent the unambiguous identification of plant water origin using water isotopes. We explored the causes of this isotopic offset by conducting a controlled experiment on the temperate tree species Fagus sylvatica. We measured δ2 H and δ18 O of soil and stem water from potted saplings growing on three soil substrates and subjected to two watering regimes. Regardless of substrate, soil and stem water δ2 H were similar only near permanent wilting point. Under moister conditions, stem water δ2 H was 11 ± 3‰ more negative than soil water δ2 H, coherent with field studies. Under drier conditions, stem water δ2 H became progressively more enriched than soil water δ2 H. Although stem water δ18 O broadly reflected that of soil water, soil-stem δ2 H and δ18 O differences were correlated (r = 0.76) and increased with transpiration rates indicated by proxies. Soil-stem isotopic offsets are more likely to be caused by water isotope heterogeneities within the soil pore and stem tissues, which would be masked under drier conditions as a result of evaporative enrichment, than by fractionation under root water uptake. Our results challenge our current understanding of isotopic signals in the soil-plant continuum.

A novel optimization approach incorporating non-stomatal limitations predicts stomatal behaviour in species from six plant functional types.

The primary function of stomata is to minimize plant water loss while maintaining CO2 assimilation. Stomatal water loss incurs an indirect cost to photosynthesis in the form of non-stomatal limitations (NSL) via reduced carboxylation capacity (CAP) and/or mesophyll conductance (MES). Two optimal formulations for stomatal conductance (gs) arise from the assumption of each type of NSL. In reality, both NSL could coexist, but one may prevail for a given leaf ontogenetic stage or plant functional type, depending on leaf morphology. We tested the suitability of two gs formulations (CAP versus MES) on species from six plant functional types (C4 crop, C3 grass, fern, conifer, evergreen, and deciduous angiosperm trees). MES and CAP parameters (the latter proportional to the marginal water cost to carbon gain) decreased with water availability only in deciduous angiosperm trees, while there were no clear differences between leaf ontogenetic stages. Both CAP and MES formulations fit our data in most cases, particularly under low water availability. For ferns, stomata appeared to operate optimally only when subjected to water stress. Overall, the CAP formulation provided a better fit across all species, suggesting that sub-daily stomatal responses minimize NSL by reducing carboxylation capacity predominantly, regardless of leaf morphology and ontogenetic stage.

Soil exchange rates of COS and CO18O differ with the diversity of microbial communities and their carbonic anhydrase enzymes.

Differentiating the contributions of photosynthesis and respiration to the global carbon cycle is critical for improving predictive climate models. Carbonic anhydrase (CA) activity in leaves is responsible for the largest biosphere-atmosphere trace gas fluxes of carbonyl sulfide (COS) and the oxygen-18 isotopologue of carbon dioxide (CO18O) that both reflect gross photosynthetic rates. However, CA activity also occurs in soils and will be a source of uncertainty in the use of COS and CO18O as carbon cycle tracers until process-based constraints are improved. In this study, we measured COS and CO18O exchange rates and estimated the corresponding CA activity in soils from a range of biomes and land use types. Soil CA activity was not uniform for COS and CO2, and patterns of divergence were related to microbial community composition and CA gene expression patterns. In some cases, the same microbial taxa and CA classes catalyzed both COS and CO2 reactions in soil, but in other cases the specificity towards the two substrates differed markedly. CA activity for COS was related to fungal taxa and ß-D-CA expression, whereas CA activity for CO2 was related to algal and bacterial taxa and α-CA expression. This study integrates gas exchange measurements, enzyme activity models, and characterization of soil taxonomic and genetic diversity to build connections between CA activity and the soil microbiome. Importantly, our results identify kinetic parameters to represent soil CA activity during application of COS and CO18O as carbon cycle tracers.

Estimating Mesophyll Conductance from Measurements of C18OO Photosynthetic Discrimination and Carbonic Anhydrase Activity.

Carbonic anhydrase (CA) activity in leaves catalyzes the 18O exchange between CO2 and water during photosynthesis. This feature has been used to estimate the mesophyll conductance to CO2 (g m) from measurements of online C18OO photosynthetic discrimination (∆18O). Based on CA assays on leaf extracts, it has been argued that CO2 in mesophyll cells should be in isotopic equilibrium with water in most C3 species as well as many C4 dicot species. However, this seems incompatible with ∆18O data that would indicate a much lower degree of equilibration, especially in C4 plants under high light intensity. This apparent contradiction is resolved here using a new model of C3 and C4 photosynthetic discrimination that includes competition between CO2 hydration and carboxylation and the contribution of respiratory fluxes. This new modeling framework is used to revisit previously published data sets on C3 and C4 species, including CA-deficient plants. We conclude that (1) newly ∆18O-derived g m values are usually close but significantly higher (typically 20% and up to 50%) than those derived assuming full equilibration and (2) despite the uncertainty associated with the respiration rate in light, or the water isotope gradient between mesophyll and bundle sheath cells, robust estimates of ∆18O-derived g m can be achieved in both C3 and C4 plants.

The interaction of soil phototrophs and fungi with pH and their impact on soil CO2, CO18O and OCS exchange.

The stable oxygen isotope composition of atmospheric CO2 and the mixing ratio of carbonyl sulphide (OCS) are potential tracers of biospheric CO2 fluxes at large scales. However, the use of these tracers hinges on our ability to understand and better predict the activity of the enzyme carbonic anhydrase (CA) in different soil microbial groups, including phototrophs. Because different classes of the CA family (α, ß and γ) may have different affinities to CO2 and OCS and their expression should also vary between different microbial groups, differences in the community structure could impact the 'community-integrated' CA activity differently for CO2 and OCS. Four soils of different pH were incubated in the dark or with a diurnal cycle for forty days to vary the abundance of native phototrophs. Fluxes of CO2, CO18O and OCS were measured to estimate CA activity alongside the abundance of bacteria, fungi and phototrophs. The abundance of soil phototrophs increased most at higher soil pH. In the light, the strength of the soil CO2 sink and the CA-driven CO2-H2O isotopic exchange rates correlated with phototrophs abundance. OCS uptake rates were attributed to fungi whose abundance was positively enhanced in alkaline soils but only in the presence of increased phototrophs. Our findings demonstrate that soil-atmosphere CO2, OCS and CO18O fluxes are strongly regulated by the microbial community structure in response to changes in soil pH and light availability and supports the idea that different members of the microbial community express different classes of CA, with different affinities to CO2 and OCS.

How do leaf and ecosystem measures of water-use efficiency compare?

The terrestrial carbon and water cycles are intimately linked: the carbon cycle is driven by photosynthesis, while the water balance is dominated by transpiration, and both fluxes are controlled by plant stomatal conductance. The ratio between these fluxes, the plant water-use efficiency (WUE), is a useful indicator of vegetation function. WUE can be estimated using several techniques, including leaf gas exchange, stable isotope discrimination, and eddy covariance. Here we compare global compilations of data for each of these three techniques. We show that patterns of variation in WUE across plant functional types (PFTs) are not consistent among the three datasets. Key discrepancies include the following: leaf-scale data indicate differences between needleleaf and broadleaf forests, but ecosystem-scale data do not; leaf-scale data indicate differences between C3 and C4 species, whereas at ecosystem scale there is a difference between C3 and C4 crops but not grasslands; and isotope-based estimates of WUE are higher than estimates based on gas exchange for most PFTs. Our study quantifies the uncertainty associated with different methods of measuring WUE, indicates potential for bias when using WUE measures to parameterize or validate models, and indicates key research directions needed to reconcile alternative measures of WUE.

Bryophyte gas-exchange dynamics along varying hydration status reveal a significant carbonyl sulphide (COS) sink in the dark and COS source in the light.

Carbonyl sulphide (COS) is a potential tracer of gross primary productivity (GPP), assuming a unidirectional COS flux into the vegetation that scales with GPP. However, carbonic anhydrase (CA), the enzyme that hydrolyses COS, is expected to be light independent, and thus plants without stomata should continue to take up COS in the dark. We measured net CO2 (AC ) and COS (AS ) uptake rates from two astomatous bryophytes at different relative water contents (RWCs), COS concentrations, temperatures and light intensities. We found large AS in the dark, indicating that CA activity continues without photosynthesis. More surprisingly, we found a nonzero COS compensation point in light and dark conditions, indicating a temperature-driven COS source with a Q10 (fractional change for a 10°C temperature increase) of 3.7. This resulted in greater AS in the dark than in the light at similar RWC. The processes underlying such COS emissions remain unknown. Our results suggest that ecosystems dominated by bryophytes might be strong atmospheric sinks of COS at night and weaker sinks or even sources of COS during daytime. Biotic COS production in bryophytes could result from symbiotic fungal and bacterial partners that could also be found on vascular plants.

A dynamic leaf gas-exchange strategy is conserved in woody plants under changing ambient CO2 : evidence from carbon isotope discrimination in paleo and CO2 enrichment studies.

Rising atmospheric [CO2 ], ca , is expected to affect stomatal regulation of leaf gas-exchange of woody plants, thus influencing energy fluxes as well as carbon (C), water, and nutrient cycling of forests. Researchers have proposed various strategies for stomatal regulation of leaf gas-exchange that include maintaining a constant leaf internal [CO2 ], ci , a constant drawdown in CO2 (ca  - ci ), and a constant ci /ca . These strategies can result in drastically different consequences for leaf gas-exchange. The accuracy of Earth systems models depends in part on assumptions about generalizable patterns in leaf gas-exchange responses to varying ca . The concept of optimal stomatal behavior, exemplified by woody plants shifting along a continuum of these strategies, provides a unifying framework for understanding leaf gas-exchange responses to ca . To assess leaf gas-exchange regulation strategies, we analyzed patterns in ci inferred from studies reporting C stable isotope ratios (δ(13) C) or photosynthetic discrimination (∆) in woody angiosperms and gymnosperms that grew across a range of ca spanning at least 100 ppm. Our results suggest that much of the ca -induced changes in ci /ca occurred across ca spanning 200 to 400 ppm. These patterns imply that ca  - ci will eventually approach a constant level at high ca because assimilation rates will reach a maximum and stomatal conductance of each species should be constrained to some minimum level. These analyses are not consistent with canalization toward any single strategy, particularly maintaining a constant ci . Rather, the results are consistent with the existence of a broadly conserved pattern of stomatal optimization in woody angiosperms and gymnosperms. This results in trees being profligate water users at low ca , when additional water loss is small for each unit of C gain, and increasingly water-conservative at high ca , when photosystems are saturated and water loss is large for each unit C gain.

Biophysical modelling of intra-ring variations in tracheid features and wood density of Pinus pinaster trees exposed to seasonal droughts.

Process-based models that link seasonally varying environmental signals to morphological features within tree rings are essential tools to predict tree growth response and commercially important wood quality traits under future climate scenarios. This study evaluated model portrayal of radial growth and wood anatomy observations within a mature maritime pine (Pinus pinaster (L.) Aït.) stand exposed to seasonal droughts. Intra-annual variations in tracheid anatomy and wood density were identified through image analysis and X-ray densitometry on stem cores covering the growth period 1999-2010. A cambial growth model was integrated with modelled plant water status and sugar availability from the soil-plant-atmosphere transfer model MuSICA to generate estimates of cell number, cell volume, cell mass and wood density on a weekly time step. The model successfully predicted inter-annual variations in cell number, ring width and maximum wood density. The model was also able to predict the occurrence of special anatomical features such as intra-annual density fluctuations (IADFs) in growth rings. Since cell wall thickness remained surprisingly constant within and between growth rings, variations in wood density were primarily the result of variations in lumen diameter, both in the model and anatomical data. In the model, changes in plant water status were identified as the main driver of the IADFs through a direct effect on cell volume. The anatomy data also revealed that a trade-off existed between hydraulic safety and hydraulic efficiency. Although a simplified description of cambial physiology is presented, this integrated modelling approach shows potential value for identifying universal patterns of tree-ring growth and anatomical features over a broad climatic gradient.

Carbon isotope discrimination during branch photosynthesis of Fagus sylvatica: field measurements using laser spectrometry.

On-line measurements of photosynthetic carbon isotope discrimination ((13)Δ) under field conditions are sparse. Hence, experimental verification of the natural variability of instantaneous (13)Δ is scarce, although (13)Δ is, explicitly and implicitly, used from leaf to global scales for inferring photosynthetic characteristics. This work presents the first on-line field measurements of (13)Δ of Fagus sylvatica branches, at hourly resolution, using three open branch bags and a laser spectrometer for CO2 isotopologue measurements (QCLAS-ISO). Data from two August/September field campaigns, in 2009 and 2010, in a temperate forest in Switzerland are shown. Diurnal variability of (13)Δ was substantial, with mean diurnal amplitudes of ~9‰ and maximum diurnal amplitudes of ~20‰. The highest (13)Δ were generally observed during early morning and late afternoon, and the lowest (13)Δ during midday. An assessment of propagated standard deviations of (13)Δ demonstrated that the observed diurnal variation of (13)Δ was not a measurement artefact. Day-to-day variations of (13)Δ were summarized with flux-weighted daily means of (13)Δ, which ranged from 15‰ to 23‰ in 2009 and from 18‰ to 29‰ in 2010, thus displaying a considerable range of 8-11‰. Generally, (13)Δ showed the expected negative relationship with intrinsic water use efficiency. Diurnal and day-to-day variability of (13)Δ was, however, always better predicted by that of net CO2 assimilation, especially in 2010 when soil moisture was high and vapour pressure deficit was low. Stomatal control of leaf gas exchange, and consequently (13)Δ, could only be identified under drier conditions in 2009.

Carbon isotope discrimination during branch photosynthesis of Fagus sylvatica: a Bayesian modelling approach.

Field measurements of photosynthetic carbon isotope discrimination ((13)Δ) of Fagus sylvatica, conducted with branch bags and laser spectrometry, revealed a high variability of (13)Δ, both on diurnal and day-to-day timescales. We tested the prediction capability of three versions of a commonly used model for (13)Δ [called here comprehensive ((13)(Δcomp)), simplified ((13) Δsimple) and revised ((13)(Δrevised)) versions]. A Bayesian approach was used to calibrate major model parameters. Constrained estimates were found for the fractionation during CO(2) fixation in (13)(Δcomp), but not in (13)(Δsimple), and partially for the mesophyll conductance for CO(2)(gi). No constrained estimates were found for fractionations during mitochondrial and photorespiration, and for a diurnally variable apparent fractionation between current assimilates and mitochondrial respiration, specific to (13)(Δrevised). A quantification of parameter estimation uncertainties and interdependencies further helped explore model structure and behaviour. We found that (13)(Δcomp) usually outperformed (13)(Δsimple) because of the explicit consideration of gi and the photorespiratory fractionation in (13)(Δcomp) that enabled a better description of the large observed diurnal variation (≈9‰) of (13)Δ. Flux-weighted daily means of (13)Δ were also better predicted with (13)(Δcomp) than with (13)(Δsimple).