Post-photosynthetic Fractionation Research Articles

Why leaves become isotopically lighter than photosynthetic carbon isotope discrimination explains: on the importance of post-photosynthetic fractionation.

This article comments on: Yu YZ, Liu HT, Yang F, Li L, Schäufele R, Tcherkez G, Schnyder H, Gong XY. 2024. δ13C of bulk organic matter and cellulose reveal post-photosynthetic fractionation during ontogeny in C4 grass leaves. Journal of Experimental Botany 75, 1451–1464

δ13C of bulk organic matter and cellulose reveal post-photosynthetic fractionation during ontogeny in C4 grass leaves.

The 13C isotope composition (δ13C) of leaf dry matter is a useful tool for physiological and ecological studies. However, how post-photosynthetic fractionation associated with respiration and carbon export influences δ13C remains uncertain. We investigated the effects of post-photosynthetic fractionation on δ13C of mature leaves of Cleistogenes squarrosa, a perennial C4 grass, in controlled experiments with different levels of vapour pressure deficit and nitrogen supply. With increasing leaf age class, the 12C/13C fractionation of leaf organic matter relative to the δ13C of atmosphere CO2 (ΔDM) increased while that of cellulose (Δcel) was almost constant. The divergence between ΔDM and Δcel increased with leaf age class, with a maximum value of 1.6‰, indicating the accumulation of post-photosynthetic fractionation. Applying a new mass balance model that accounts for respiration and export of photosynthates, we found an apparent 12C/13C fractionation associated with carbon export of -0.5‰ to -1.0‰. Different ΔDM among leaves, pseudostems, daughter tillers, and roots indicate that post-photosynthetic fractionation happens at the whole-plant level. Compared with ΔDM of old leaves, ΔDM of young leaves and Δcel are more reliable proxies for predicting physiological parameters due to the lower sensitivity to post-photosynthetic fractionation and the similar sensitivity in responses to environmental changes.

Drivers of intra-seasonal δ13 C signal in tree-rings of Pinussylvestris as indicated by compound-specific and laser ablation isotope analysis.

Carbon isotope composition of tree-ring (δ13 CRing ) is a commonly used proxy for environmental change and ecophysiology. δ13 CRing reconstructions are based on a solid knowledge of isotope fractionations during formation of primary photosynthates (δ13 CP ), such as sucrose. However, δ13 CRing is not merely a record of δ13 CP . Isotope fractionation processes, which are not yet fully understood, modify δ13 CP during sucrose transport. We traced, how the environmental intra-seasonal δ13 CP signal changes from leaves to phloem, tree-ring and roots, for 7year old Pinus sylvestris, using δ13 C analysis of individual carbohydrates, δ13 CRing laser ablation, leaf gas exchange and enzyme activity measurements. The intra-seasonal δ13 CP dynamics was clearly reflected by δ13 CRing , suggesting negligible impact of reserve use on δ13 CRing . However, δ13 CP became increasingly 13 C-enriched during down-stem transport, probably due to post-photosynthetic fractionations such as sink organ catabolism. In contrast, δ13 C of water-soluble carbohydrates, analysed for the same extracts, did not reflect the same isotope dynamics and fractionations as δ13 CP , but recorded intra-seasonal δ13 CP variability. The impact of environmental signals on δ13 CRing , and the 0.5and 1.7‰ depletion in photosynthates compared ring organic matter and tree-ring cellulose, respectively, are useful pieces of information for studies exploiting δ13 CRing .

Estimation of intrinsic water-use efficiency from δ13C signature of C3 leaves: Assumptions and uncertainty.

Carbon isotope composition (δ13C) has been widely used to estimate the intrinsic water-use efficiency (iWUE) of plants in ecosystems around the world, providing an ultimate record of the functional response of plants to climate change. This approach relies on established relationships between leaf gas exchange and isotopic discrimination, which are reflected in different formulations of 13C-based iWUE models. In the current literature, most studies have utilized the simple, linear equation of photosynthetic discrimination to estimate iWUE. However, recent studies demonstrated that using this linear model for quantitative studies of iWUE could be problematic. Despite these advances, there is a scarcity of review papers that have comprehensively reviewed the theoretical basis, assumptions, and uncertainty of 13C-based iWUE models. Here, we 1) present the theoretical basis of 13C-based iWUE models: the classical model (iWUEsim), the comprehensive model (iWUEcom), and the model incorporating mesophyll conductance (iWUEmes); 2) discuss the limitations of the widely used iWUEsim model; 3) and make suggestions on the application of the iWUEmes model. Finally, we suggest that a mechanistic understanding of mesophyll conductance associated effects and post-photosynthetic fractionation are the bottlenecks for improving the 13C-based estimation of iWUE.

δ13C in above-ground and below-ground organs of Spinulum annotinum (Lycopodiaceae)

• 12 C and 13 C isotope abundance in S. annotinum has been investigated for the first time. • δ 13 C value in S. annotinum is lower than that reported in other C 3 plants. • Different above- and below-ground S. annotinum organs have distinct δ 13 C values. • δ 13 C values of S. annotinum are connected with local plant diversity. Physiological processes in plants and plant–environment interactions can be analysed from the composition of stable carbon isotopes ( 13 C/ 12 C). This ratio can be indicated using the δ (delta) notation. Substantial evidence has been published in recent years demonstrating that post-photosynthetic fractionation occurs in plants and leads to differences in 13 C-enrichment in heterotrophic (as compared with autotrophic) organs. Yet our understanding of processes leading to 13 C divergence between leaves and heterotrophic tissues remains low. Large part of the isotope analysis is concerned with angiosperms, but not spore-bearing tracheophytes. In this study, we explored the 12 C and 13 C isotope distribution in above-ground and below-ground organs of an archaic lycophyte Spinulum annotinum (L.) A.Haines (interrupted clubmoss). The main objectives of this study were to determine the natural distribution of stable carbon isotopes in sporophytes of S. annotinum and to discuss possible causes for differences in δ 13 C. The study revealed that there are significant differences between δ 13 C in different S. annotinum organs. The δ 13 C values varied by about 2.4‰. The highest δ 13 C values were −30.0‰ in roots and the lowest were −32.4‰ in plagiotropic shoot leaves. The distance to a body of water had no significant effect on δ 13 C values in S. annotinum . Outgrowth size did not have a clear relationship with δ 13 C in S. annotinum . The Shannon Diversity Index was a significant predictor for δ 13 C values in S. annotinum . The uncovered δ 13 C and the relationship with abiotic habitat conditions and neighbouring plant species in the community provide a base for future lycophyte ecophysiology research.

Species variation in the hydrogen isotope composition of leaf cellulose is mostly driven by isotopic variation in leaf sucrose.

Experimental approaches to isolate drivers of variation in the carbon-bound hydrogen isotope composition (δ2 H) of plant cellulose are rare and current models are limited in their application. This is in part due to a lack in understanding of how 2 H-fractionations in carbohydrates differ between species. We analysed, for the first time, the δ2 H of leaf sucrose along with the δ2 H and δ18 O of leaf cellulose and leaf and xylem water across seven herbaceous species and a starchless mutant of tobacco. The δ2 H of sucrose explained 66% of the δ2 H variation in cellulose (R2 = 0.66), which was associated with species differences in the 2 H enrichment of sucrose above leaf water ( : -126% to -192‰) rather than by variation in leaf water δ2 H itself. was positively related to dark respiration (R2 = 0.27), and isotopic exchange of hydrogen in sugars was positively related to the turnover time of carbohydrates (R2 = 0.38), but only when was fixed to the literature accepted value of ‰. No relation was found between isotopic exchange of hydrogen and oxygen, suggesting large differences in the processes shaping post-photosynthetic fractionation between elements. Our results strongly advocate that for robust applications of the leaf cellulose hydrogen isotope model, parameterization utilizing δ2 H of sugars is needed.

Whole-tree mesophyll conductance reconciles isotopic and gas-exchange estimates of water-use efficiency.

Photosynthetic water-use efficiency (WUE) describes the link between terrestrial carbon (C) and water cycles. Estimates of intrinsic WUE (iWUE) from gas exchange and C isotopic composition (δ13 C) differ due to an internal conductance in the leaf mesophyll (gm ) that is variable and seldom computed. We present the first direct estimates of whole-tree gm , together with iWUE from whole-tree gas exchange and δ13 C of the phloem (δ13 Cph ). We measured gas exchange, online 13 C-discrimination, and δ13 Cph monthly throughout spring, summer, and autumn in Eucalyptus tereticornis grown in large whole-tree chambers. Six trees were grown at ambient temperatures and six at a 3°C warmer air temperature; a late-summer drought was also imposed. Drought reduced whole-tree gm . Warming had few direct effects, but amplified drought-induced reductions in whole-tree gm . Whole-tree gm was similar to leaf gm for these same trees. iWUE estimates from δ13 Cph agreed with iWUE from gas exchange, but only after incorporating gm . δ13 Cph was also correlated with whole-tree 13 C-discrimination, but offset by -2.5±0.7‰, presumably due to post-photosynthetic fractionations. We conclude that δ13 Cph is a good proxy for whole-tree iWUE, with the caveats that post-photosynthetic fractionations and intrinsic variability of gm should be incorporated to provide reliable estimates of this trait in response to abiotic stress.

Effects of soil water stress and atmospheric CO2 concentration on photosynthetic and post-photosynthetic fractionation

Analysis of plant photosynthesis and post-photosynthetic fractionation can improve our understanding of plant physiology and water management. By measuring δ13C in the atmosphere, and δ13C of soluble compounds in leaves and branch phloem of Platycladus orientalis, we examined discrimination pattern, including atmosphere-leaf discrimination during photosynthesis (ΔCa-leaf) and leaf-twig discrimination during post-photosynthesis (ΔCleaf-phlo), in response to changes of soil water content (SWC) and atmospheric CO2 concentration (Ca). The results showed that ΔCa-leaf reached a maximum of 13.06‰ at 95%-100% field water-holding capacity (FC) and Ca 400 μmol·mol-1, and a minimum of 8.63‰ at 35%-45% FC and Ca 800 μmol·mol-1. Both stomatal conductance and mesophyll cell conductance showed a significant linear positive correlation with ΔCa-leaf, with a correlation coefficient of 0.43 and 0.44, respectively. ΔCleaf-phlo was not affected by SWC and Ca. Our results provide mechanism of carbon isotopes fractionation and a theoretical basis for plant survival strategies in response to future climate change.

A Comparison of the Variable J and Carbon-Isotopic Composition of Sugars Methods to Assess Mesophyll Conductance from the Leaf to the Canopy Scale in Drought-Stressed Cherry.

Conductance of CO2 across the mesophyll (Gm) frequently constrains photosynthesis (PN) but cannot be measured directly. We examined Gm of cherry (Prunus avium L.) subjected to severe drought using the variable J method and carbon-isotopic composition (δ13C) of sugars from the centre of the leaf, the leaf petiole sap, and sap from the largest branch. Depending upon the location of the plant from which sugars are sampled, Gm may be estimated over scales ranging from a portion of the leaf to a canopy of leaves. Both the variable J and δ13C of sugars methods showed a reduction in Gm as soil water availability declined. The δ13C of sugars further from the source of their synthesis within the leaf did not correspond as closely to the diffusive and C-isotopic discrimination conditions reflected in the instantaneous measurement of gas exchange and chlorophyll-fluorescence utilised by the variable J approach. Post-photosynthetic fractionation processes and/or the release of sugars from stored carbohydrates (previously fixed under different environmental and C-isotopic discrimination conditions) may reduce the efficacy of the δ13C of sugars from leaf petiole and branch sap in estimating Gm in a short-term study. Consideration should be given to the spatial and temporal scales at which Gm is under observation in any experimental analysis.

Variations in δ13C of water–soluble leaf and phloem organic matter of Platycladus orientalis: influences of photosynthetic and post–photosynthetic fractionation

ABSTRACTAnalysis of variations in water–soluble organic matter (WSOM) δ13C of leaves and phloem can efficiently describe the δ13C distributions within plants and identify the temporal variation of δ13C. In this study, WSOM δ13C values of both leaves and phloem (twig, stem, and root) of Platycladus orientalis were measured during seven sunny days, including 2–hour interval measurements at three days for diel pattern analysis and 6–hour interval measurements at the remaining four days for day–to–day variation analysis. Analysis of WSOM δ13C in different plant organs showed that 13C was generally depleted from leaves to twigs, then enriched in stems and subsequently depleted in roots. Stems were significantly 13C–enriched compared to twigs (p < 0.05), while δ13C differences between stems and other organs and among leaves, twigs and roots were not significant (p > 0.05). No clear diel patterns in δ13C of leaves and phloem were found. Daily average δ13C values indicated that all plant organs had more positive values on sunny days during the dry season than during the wet season. Both photosynthetic and post–photosynthetic fractionation influence variations in WSOM δ13C. These results have implications for research on plant physiology and plant water use.

Biochemical and transcriptomic analysis of maize diversity to elucidate drivers of leaf carbon isotope composition.

Carbon isotope discrimination is used to study CO2 diffusion, substrate availability for photosynthesis, and leaf biochemistry, but the intraspecific drivers of leaf carbon isotope composition (δ13C) in C4 species are not well understood. In this study, the role of photosynthetic enzymes and post-photosynthetic fractionation on δ13C (‰) was explored across diverse maize inbred lines. A significant 1.3‰ difference in δ13C was observed between lines but δ13C did not correlate with in vitro leaf carbonic anhydrase (CA), phosphoenolpyruvate carboxylase (PEPC), or ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity. RNA-sequencing was used to identify potential differences in post-photosynthetic metabolism that would influence δ13C; however, no correlations were identified that would indicate significant differences in post-photosynthetic fractionation between lines. Variation in δ13C has been observed between C4 subtypes, but differential expression of NADP-ME and PEP-CK pathways within these lines did not correlate with δ13C. However, co-expression network analysis provided novel evidence for isoforms of C4 enzymes and putative transporters. Together, these data indicate that diversity in maize δ13C cannot be fully explained by variation in CA, PEPC, or Rubisco activity or gene expression. The findings further emphasise the need for future work exploring the influence of stomatal sensitivity and mesophyll conductance on δ13C in maize.

Stress-induced changes in carbon allocation among metabolite pools influence isotope-based predictions of water use efficiency in Phaseolus vulgaris.

Understanding how major food crops respond to environmental stress will expand our capacity to improve food production with growing populations and a changing climate. This study uses chemical and physiological adaptations to heat, water deficit and elevated light stresses in Phaseolus vulgaris L. to identify changes in carbon (C) allocation that, combined with post-photosynthetic fractionation of C isotopes, influences water use efficiency (WUE) predictions. The chemical stress response was explored through changes in C allocation to the carbohydrate and cyclitol pools using GC-triple quadrupole MS. Carbon allocation to the sucrose pool fluctuated significantly among treatments, and the putative osmolytes and osmoprotectants (myo-inositol and d-ononitol) accumulated under stress. Significant osmotic adjustment (P<0.05), quantified via pressure-volume curve analysis, was detected between control and stress treatments, although this was not attributable to active accumulation of the metabolites. Compound-specific 13C isotope abundance was measured using liquid chromatography isotope ratio MS to predict intrinsic WUE. In contrast to other metabolites measured, the δ13C of the sucrose pool fluctuated according to treatment and was proportional to predicted values based upon modelled Δ13C from gas exchange data. The results suggest that the accuracy and precision of predicting WUE may be enhanced by compound-specific analysis of Δ13C and that changes in the allocation of C among metabolite pools may influence WUE predictions based upon analysis of total soluble C. Overall, the plants appeared to use a range of mechanisms to cope with adverse conditions that could be utilised to improve plant breeding and management strategies.

Preface &amp;quot;Stable Isotopes and Biogeochemical Cycles in Terrestrial Ecosystems''

In view of a rapidly changing environment there is the need to better understand the response of ecosystems to global changes of climate and land use. While there are substantial ongoing efforts in monitoring fluxes of carbon (C), water and nitrogen (N) between ecosystems and their environment (e.g. Baldocchi, 2008; Sutton et al., 2012), there is still an insufficient understanding of the processes underpinning biogeochemical cycles at the interfaces between atmosphere, plant, soil and microbes. One important field that deserves increasing attention concerns plant – soil interactions (see also recent special issue, Subke et al., 2012), whose study has so far been limited by considerable methodological problems, in particular in terms of the soil compartment. Stable isotopes are a powerful tool for tracing elements and for unravelling plant and soil processes as well as their coupling to the atmosphere at various temporal and spatial scales. Stable isotopes have yielded significant breakthroughs, such as identifying the imprint of biospheric CO2 fluxes from terrestrial ecosystems on the atmosphere (using13C and 18O), separating autotrophic and heterotrophic respiration in soils (13C) and quantifying atmospheric N2 inputs to ecosystems (15N) and their impact on ecosystem functions (Ciais et al., 1995; Yakir and Sternberg, 2000; Robinson, 2001; Flanagan et al., 2005; Dawson and Siegwolf, 2007; Bowling et al., 2008). Over the last decade, advances in measurement techniques, e.g. laser spectrometry and compound specific analysis, have enabled scientists to apply stable isotopes to study processes in environmental research in so far unprecedented resolution and detail. International efforts to network scientists applying stable isotopes in different earth system science disciplines, such as COST Action ES0806 Stable Isotopes in BiosphereAtmosphere-Earth System Research (SIBAE), resulted in further advancements of our mechanistic understanding. This special issue on “Stable Isotopes and Biogeochemical Cycles in Terrestrial Ecosystems” highlights some of these recent advances and presents reviews and case studies addressing the following topics: (1) tracing C from photosynthetic assimilation to respired CO2, soil organic matter and soil biota, (2) unraveling and upscaling nitrogen dynamics, (3) analyzing linkages between carbon and water cycles, and (4) applying isotopes for constraining global biogeochemical models. C allocation is a key process in terrestrial ecosystems, whose dynamics are still poorly understood but can now be studied in some detail using isotopic tracers combined with compound-specific analysis and/or isotope laser spectroscopy (Bruggemann et al., 2011; Epron et al., 2012). The natural stable carbon and oxygen isotope composition of respiratory substrates and of plantand soil-respired CO2 follows pronounced diel variations, which typically decrease from leaves to trunks and roots (Gavrichkova et al., 2011; Werner and Gessler, 2011). These are most likely determined by post-photosynthetic fractionation processes as related to changes in C allocation to different metabolic pathways, and by a mixing of substrates and of component fluxes (Werner and Gessler, 2011). In situ pulse labeling experiments on mature trees indicate that the patterns of C allocation to belowground respiration are species-specific and change seasonally depending on the phenology of species (Epron et al., 2011). Stress, e.g. induced by chronic ozone exposure, can reduce the allocation of recent photosynthates to stem and root respiration (Ritter et al., 2011; cf. also Bruggemann et al., 2011). Long-term tracer application of CO2 by free air CO2 enrichment (FACE) permits identifying the diurnal

Short-term natural δ&amp;lt;sup&amp;gt;13&amp;lt;/sup&amp;gt;C and δ&amp;lt;sup&amp;gt;18&amp;lt;/sup&amp;gt;O variations in pools and fluxes in a beech forest: the transfer of isotopic signal from recent photosynthates to soil respired CO&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt;

Abstract. The fate of photosynthetic products within the plant-soil continuum determines how long the reduced carbon resides within the ecosystem and when it returns back to the atmosphere in the form of respiratory CO2. We have tested the possibility of measuring natural variation in δ13C and δ18O to disentangle the potential times needed to transfer carbohydrates produced by photosynthesis down to trunk, roots and, in general, to belowground up to its further release in the form of soil respiration into the atmosphere in a beech (Fagus sylvatica) forest. We have measured the variation in stable carbon and oxygen isotope compositions in plant material and in soil respired CO2 every three hours for three consecutive days. Possible steps and different signs of post-photosynthetic fractionation during carbon translocation were also identified. A 12 h-periodicity was observed for variation in δ13C in soluble sugars in the top crown leaves and it can be explained by starch day/night dynamics in synthesis and breakdown and by stomatal limitations under elevated vapour pressure deficits. Photosynthetic products were transported down the trunk and mixed with older carbon pools, therefore causing the dampening of the δ13C signal variation. The strongest periodicity of 24 h was found in δ13C in soil respiration indicating changes in root contribution to the total CO2 efflux. Other non-biological causes like diffusion fractionation and advection induced by gas withdrawn from the measurement chamber complicate data interpretation on this step of C transfer path. Nevertheless, it was possible to identify the speed of carbohydrates' translocation from the point of assimilation to the trunk breast height because leaf-imprinted enrichment of δ18O in soluble sugars was less modified along the downward transport and was well related to environmental parameters potentially linked to stomatal conductance. The speed of carbohydrates translocation from the site of assimilation to the trunk at breast height was estimated to be in the range of 0.3–0.4 m h−1.

Variation in bulk tissue, fatty acid and monosaccharide δ13C values between autotrophic and heterotrophic plant organs

The flowers of 23 species of grass and herb plants were collected from a mesotrophic grassland to assess natural variability in bulk, monosaccharide and fatty acid δ 13C values from one plant community and were compared with previous analyses of leaves from the same species. The total mean bulk δ 13C value of flower tissues was −28.1‰, and there was no significant difference between the mean δ 13C flower values for grass (−27.8‰) and herb (−28.2‰) species. On average bulk δ 13C flower values were 1.1‰ higher than bulk δ 13C leaf values, however, the δ 13C flower and δ 13C leaf values of grasses did not differ between organs suggesting that carbon isotope discrimination is different in grass and herb species. The abundance of different monosaccharides abundance varied between plant types, i.e. xylose concentrations in the grass flowers were as high as 40%, compared with up to 15% in the herb species, but the general relationship δ 13C arabinose > δ 13C xylose > δ 13C glucose > δ 13C galactose which had been observed in leaves was similar in flowers (total mean δ 13C values = −25.9‰, −27.2‰, −28.8‰ and −28.1‰, respectively). However, the average 5.4‰ depletion in the δ 13C values of the C 16:0, C 18:2 and C 18:3 fatty acids in flowers compared to bulk tissue was significantly greater than observed for leaves. The trend C 16:0 < C 18:2 < C 18:3 previously observed in leaves was also observed in grass flowers ( δ 13C C16:0 = −33.8‰; δ 13C C18:2 = −33.1‰; δ 13C C18:3 = −34.2‰) but not herb flowers ( δ 13C C16:0 = −34.1‰; δ 13C C18:2 = −32.4‰; δ 13C C18:3 = −34.5‰). We conclude: (i) that the biological processes influencing carbon isotope discrimination in grass flowers are different from herbs flowers; and, (ii) that a range of post-photosynthetic fractionation effects caused the observed differences between flower and leaf δ 13C values, especially the significant 13C-depletion in flower fatty acid δ 13C values.

On the use of phloem sap 13C as an indicator of canopy carbon discrimination

In this study we measured δ¹³C in various carbon pools along the basipetal transport pathway in co-occurring Pinus pinaster and Acacia longifolia trees under Mediterranean climate conditions in the field. Overall, species differences in photosynthetic discrimination resulted in more enriched δ¹³C values in the water-conserving overstory P. pinaster relative to the water-spending understory invasive A. longifolia. Post-photosynthetic fractionation effects resulted in differences in δ¹³C of water-soluble organic matter pools along the plant axis with progressive depletion in δ¹³C from the canopy to the trunk (∼6.5‰ depletion in A. longifolia and ∼0.8‰ depletion in P. pinaster). Regardless of these fractionation effects, phloem sap δ¹³C in both terminal branches and the main stem correlated well with environmental parameters driving photosynthesis for both species, indicating that phloem sap δ¹³C has potential as an integrative tracer of changes in canopy carbon discrimination (Δ¹³C). Furthermore, we illustrate that a simple model based on sap flow estimated canopy stomatal conductance (G(S)) and phloem sap δ¹³C measurements has significant potential as a tool for estimating canopy-level carbon assimilation rates.

Variation of δ13C of wood and foliage with canopy height differs between evergreen and deciduous species in a temperate forest

We investigated post-photosynthetic fractionation and carbon transfer mechanisms of different plant functional types growing under the same climatic conditions in North-eastern China. The variations in δ13C of trunk and branches were compared with leaf δ13C at different canopy heights of Pinus koraiensis (evergreen coniferous species), Larix gmelinii (deciduous coniferous species) and Quercus mongolica (deciduous broad-leaved species). Results showed that δ13C of leaves increased (became more enriched) with increasing canopy height for both coniferous species (P. koraiensis, L. gmelinii) but not for Q. mongolica (a deciduous broad-leaved species). δ13C of both trunk and branches also increased with sampling height for the evergreen conifer P. koraiensis but did not significantly vary for either of the deciduous species (L. gmelinii or Q. mongolica), except a significant increase in branch δ13C for L. gmelinii. Similarly, δ13C of trunk and branches were strongly correlated with leaf δ13C only in the evergreen conifer, P. koraiensis. 13C was consistently more enriched in trunk, branches, and roots compared to leaves in all three species. Our findings suggest that, even under the same climatic conditions, different plant functional types may exhibit different carbon transfer mechanisms. This is contrary to the previous hypothesis that different carbon transfer mechanisms operate in forests of different climatic zones, especially in tropical and temperate forests. Particularly, the differences occur predominantly between evergreen and deciduous trees rather than between coniferous and broad-leaved trees. The significant difference in δ13C between leaves and wood tissues confirms a previous post-photosynthetic isotope fractionation in temperate forests.

Disentangling drought-induced variation in ecosystem and soil respiration using stable carbon isotopes

Combining C flux measurements with information on their isotopic composition can yield a process-based understanding of ecosystem C dynamics. We studied the variations in both respiratory fluxes and their stable C isotopic compositions (delta(13)C) for all major components (trees, understory, roots and soil microorganisms) in a Mediterranean oak savannah during a period with increasing drought. We found large drought-induced and diurnal dynamics in isotopic compositions of soil, root and foliage respiration (delta(13)C(res)). Soil respiration was the largest contributor to ecosystem respiration (R (eco)), exhibiting a depleted isotopic signature and no marked variations with increasing drought, similar to ecosystem respired delta(13)CO(2), providing evidence for a stable C-source and minor influence of recent photosynthate from plants. Short-term and diurnal variations in delta(13)C(res) of foliage and roots (up to 8 and 4 per thousand, respectively) were in agreement with: (1) recent hypotheses on post-photosynthetic fractionation processes, (2) substrate changes with decreasing assimilation rates in combination with increased respiratory demand, and (3) decreased phosphoenolpyruvate carboxylase activity in drying roots, while altered photosynthetic discrimination was not responsible for the observed changes in delta(13)C(res). We applied a flux-based and an isotopic flux-based mass balance, yielding good agreement at the soil scale, while the isotopic mass balance at the ecosystem scale was not conserved. This was mainly caused by uncertainties in Keeling plot intercepts at the ecosystem scale due to small CO(2) gradients and large differences in delta(13)C(res) of the different component fluxes. Overall, stable isotopes provided valuable new insights into the drought-related variations of ecosystem C dynamics, encouraging future studies but also highlighting the need of improved methodology to disentangle short-term dynamics of isotopic composition of R (eco).

Consistent patterns in leaf lamina and leaf vein carbon isotope composition across ten herbs and tree species

Wide-spread post-photosynthetic fractionation processes deplete metabolites and plant compartments in (13)C relative to assimilates to varying degrees. Fragmentation fractionation and exchange of metabolites with distinct isotopic signatures across organ boundaries further modify the patterns of plant isotopic composition. Heterotrophic organs tend to become isotopically heavier than the putative source material as a result of respiratory metabolism. In addition fractionation may occur during metabolite transport across organ and tissue boundaries. Leaf laminae, veins and petioles are leaf compartments that are arranged along a gradient of increasing weight of heterotrophic processes and along a transport chain. Thus, we expect to find consistent patterns of isotopic signatures associated with this gradient. Earlier studies on leaves of Fagus sylvatica, Glycine max, and Saccharum officinarum showed that the organic mass and cellulose of major veins or petioles were consistently more positive than the respective fraction in leaf laminae. The objective of the current study was to assess whether this pattern can be detected in a greater set of plant species. Leaves from ten species were collected in the summer of 2006 outdoors and in glasshouses. Leaf laminae including small veins were separated from the major veins and the isotopic signatures of the organic mass, and the soluble and non-soluble fractions were measured for laminae and veins separately. The organic mass, and the soluble and non-soluble fractions of leaf laminae, were depleted in (13)C relative to the veins in all cases. A general trend for the signature of organic mass being more depleted in (13)C than the soluble fraction is in accordance with well-known patterns of fractionation between metabolites.

Divergence in δ13C of dark respired CO2 and bulk organic matter occurs during the transition between heterotrophy and autotrophy in Phaseolus vulgaris plants

Substantial evidence has been published in recent years demonstrating that postphotosynthetic fractionations occur in plants, leading to (13)C-enrichment in heterotrophic (as compared with autotrophic) organs. However, less is known about the mechanism responsible for changes in these responses during plant development. The isotopic signature of both organic matter and respired CO(2) for different organs of French bean (Phaseolus vulgaris) was investigated during early ontogeny, in order to identify the developmental stage at which isotopic changes occur. Isotopic analyses of metabolites and mass balance calculations helped to constrain the metabolic processes involved. At the plant scale, apparent respiratory fractionation was constantly positive in the heterotrophic phase (c. 1 per thousand) and turned negative with autotrophy acquisition (down to -3.08 per thousand). Initially very close to that of the dry seed (-26.83 +/- 0.69 per thousand), isotopic signatures of organic matter and respired CO(2) diverged (in opposite directions) in leaves and roots after onset of photosynthesis. Respired CO(2) reached values up to -20 per thousand in leaves and became (13)C-depleted down to -29 per thousand in roots. It was concluded that isotopic differences between organs occurred subsequent to metabolic changes in the seedling during the transition from heterotrophy to autotrophy. They were especially related to respiration and respiratory fractionation.