Morphological and isotopic changes of heterocystous cyanobacteria in response to N2 partial pressure

Earth's atmospheric composition has changed significantly over geologic time. Many redox active atmospheric constituents have left evidence of their presence, while inert constituents such as dinitrogen gas (N₂) are more elusive. In this study, we examine two potential biological indicators of atmospheric N₂: the morphological and isotopic signatures of heterocystous cyanobacteria. Biological nitrogen fixation constitutes the primary source of fixed nitrogen to the global biosphere and is catalyzed by the oxygen‐sensitive enzyme nitrogenase. To protect this enzyme, some filamentous cyanobacteria restrict nitrogen fixation to microoxic cells (heterocysts) while carrying out oxygenic photosynthesis in vegetative cells. Heterocysts terminally differentiate in a pattern that is maintained as the filaments grow, and nitrogen fixation imparts a measurable isotope effect, creating two biosignatures that have previously been interrogated under modern N₂ partial pressure (pN₂) conditions. Here, we examine the effect of variable pN₂ on these biosignatures for two species of the filamentous cyanobacterium Anabaena. We provide the first in vivo estimate of the intrinsic isotope fractionation factor of Mo‐nitrogenase (ε_fix = ?2.71 ± 0.09‰) and show that, with decreasing pN₂, the net nitrogen isotope fractionation decreases for both species, while the heterocyst spacing decreases for Anabaena cylindrica and remains unchanged for Anabaena variabilis. These results are consistent with the nitrogen fixation mechanisms available in the two species. Application of these quantifiable effects to the geologic record may lead to new paleobarometric measurements for pN₂, ultimately contributing to a better understanding of Earth's atmospheric evolution.     

Seasonal variation in ammonia compensation point and nitrogen pools in beech leaves (Fagus sylvatica)

Ammonia (NH₃) fluxes between beech leaves (Fagus sylvatica) and the atmosphere were investigated in a 90-year-old forest canopy and related to leaf nitrogen (N) pools and glutamine synthetase (GS) activities. The stomatal ammonia compensation point, ?? _NH3, was measured by both a twig cuvette and bioassay techniques involving measurements of pH and ammonium (NH ₄ ⁺ ) concentration in the leaf apoplastic solution. The ?? _NH3 determined on the basis of the gas exchange measurements followed a seasonal variation with early-season peaks during leaf expansion (9.6 nmol NH₃ mol^?1 air) and late-season peaks during leaf senescence (7.3 nmol NH₃ mol^?1 air). In the mid-season, the ?? _NH3 of mature green leaves was much lower (around 3 nmol NH₃ mol^?1 air) and dropped below the NH₃ concentration in the ambient atmosphere. For comparison, ?? _NH3 obtained by the apoplastic bioassay were 7.0, 3.7 and 6.4 nmol NH₃ mol^?1 air in early-, mid-, and late -season, thus agreeing reasonably well with ?? _NH3 values derived from the gas exchange measurements. Potential NH₃ emission fluxes during early and late season were 1.31 and 0.51 nmol m^?2 leaf surface area s^?1, respectively, while leaves were a sink for NH₃ during mid-season. During leaf establishment and senescence, both apoplastic and bulk tissue NH ₄ ⁺ concentrations were relatively high coinciding with low activities of glutamine synthetase, which is a key enzyme in leaf N metabolism. In conclusion, the exchange of NH₃ between beech leaves and the atmosphere followed a seasonal variation with NH₃ emission peaks being related to N mobilization during early leaf establishment and remobilization during late leaf senescence.     

Towards a coupled paradigm of NH3‐CO2 biosphere–atmosphere exchange modelling

Stomatal conductance, one of the major plant physiological controls within NH₃ biosphere–atmosphere exchange models, is commonly estimated from semi‐empirical multiplicative schemes or simple light‐ and temperature‐response functions. However, due to their inherent parameterization on meteorological proxy variables, instead of a direct measure of stomatal opening, they are unfit for the use in climate change scenarios and of limited value for interpreting field‐scale measurements. Alternatives based on H₂O flux measurements suffer from uncertainties in the partitioning of evapotranspiration at humid sites, as well as a potential decoupling of transpiration from stomatal opening in the presence of hygroscopic particles on leaf surfaces. We argue that these problems may be avoided by directly deriving stomatal conductance from CO₂ fluxes instead. We reanalysed a data set of NH₃ flux measurements based on CO₂‐derived stomatal conductance, confirming the hypothesis that the increasing relevance of stomatal exchange with the onset of vegetation activity caused a rapid decrease of observed NH₃ deposition velocities. Finally, we argue that developing more mechanistic representations of NH₃ biosphere–atmosphere exchange can be of great benefit in many applications. These range from model‐based flux partitioning, over deposition monitoring using low‐cost samplers and inferential modelling, to a direct response of NH₃ exchange to climate change.     

Modelling non‐steady‐state isotope enrichment of leaf water in a gas‐exchange cuvette environment

The combined use of a gas‐exchange system and laser‐based isotope measurement is a tool of growing interest in plant ecophysiological studies, owing to its relevance for assessing isotopic variability in leaf water and/or transpiration under non‐steady‐state (NSS) conditions. However, the current Farquhar & Cernusak (F&C) NSS leaf water model, originally developed for open‐field scenarios, is unsuited for use in a gas‐exchange cuvette environment where isotope composition of water vapour (δ_v) is intrinsically linked to that of transpiration (δ_E). Here, we modified the F&C model to make it directly compatible with the δ_v–δ_E dynamic characteristic of a typical cuvette setting. The resultant new model suggests a role of ‘net‐flux’ (rather than ‘gross‐flux’ as suggested by the original F&C model)‐based leaf water turnover rate in controlling the time constant (τ) for the approach to steady sate. The validity of the new model was subsequently confirmed in a cuvette experiment involving cotton leaves, for which we demonstrated close agreement between τ values predicted from the model and those measured from NSS variations in isotope enrichment of transpiration. Hence, we recommend that our new model be incorporated into future isotope studies involving a cuvette condition where the transpiration flux directly influences δ_v. There is an increasing popularity among plant ecophysiologists to use a gas‐exchange system coupled to laser‐based isotope measurement for investigating non‐steady state (NSS) isotopic variability in leaf water (and/or transpiration); however, the current Farquhar & Cernusak (F&C) NSS leaf water model is unsuited for use in a gas‐exchange cuvette environment due to its implicit assumption of isotope composition of water vapor (δ_v) being constant and independent of that of transpiration (δ_E). In the present study, we modified the F&C model to make it compatible with the dynamic relationship between δ_v and δ_E as is typically associated with a cuvette setting. Using an experiment conducted on cotton leaves, we show that the modified NSS model performed well in predicting the time constant for the exponential approach of leaf water toward steady state under cuvette conditions. Such a result demonstrates the applicability of this new model to gas‐exchange cuvette conditions where the transpiration flux directly influences δ_v, and therefore suggests the need to incorporate this model into future isotope studies that employ a laser‐cuvette coupled system.     

Coupled response of stomatal and mesophyll conductance to light enhances photosynthesis of shade leaves under sunflecks

Light gradients within tree canopies play a major role in the distribution of plant resources that define the photosynthetic capacity of sun and shade leaves. However, the biochemical and diffusional constraints on gas exchange in sun and shade leaves in response to light remain poorly quantified, but critical for predicting canopy carbon and water exchange. To investigate the CO₂ diffusion pathway of sun and shade leaves, leaf gas exchange was coupled with concurrent measurements of carbon isotope discrimination to measure net leaf photosynthesis (A_n), stomatal conductance (g_s) and mesophyll conductance (g_m) in Eucalyptus tereticornis trees grown in climate controlled whole‐tree chambers. Compared to sun leaves, shade leaves had lower A_n, g_m, leaf nitrogen and photosynthetic capacity (A_max) but g_s was similar. When light intensity was temporarily increased for shade leaves to match that of sun leaves, both g_s and g_m increased, and A_n increased to values greater than sun leaves. We show that dynamic physiological responses of shade leaves to altered light environments have implications for up‐scaling leaf level measurements and predicting whole canopy carbon gain. Despite exhibiting reduced photosynthetic capacity, the rapid up‐regulation of g_m with increased light enables shade leaves to respond quickly to sunflecks.     

Strong seasonal disequilibrium measured between the oxygen isotope signals of leaf and soil CO2 exchange

The oxygen isotope composition (δ¹⁸O) of atmospheric CO₂ is among a very limited number of tools available to constrain estimates of the biospheric gross CO₂ fluxes, photosynthesis and respiration at large scales. However, the accuracy of the partitioning strongly depends on the extent of isotopic disequilibrium between the signals carried by these two gross fluxes. Chamber‐based field measurements of total CO₂ and CO¹⁸O fluxes from foliage and soil can help evaluate and refine our models of isotopic fractionation by plants and soils and validate the extent and pattern of isotopic disequilibrium within terrestrial ecosystems. Owing to sampling limitations in the past, such measurements have been very rare and covered only a few days. In this study, we coupled automated branch and soil chambers with tuneable diode laser absorption spectroscopy techniques to continuously capture the δ¹⁸O signals of foliage and soil CO₂ exchange in a Pinus pinaster Aït forest in France. Over the growing season, we observed a seasonally persistent isotopic disequilibrium between the δ¹⁸O signatures of net CO₂ fluxes from leaves and soils, except during rain events when the isotopic imbalance became temporarily weaker. Variations in the δ¹⁸O of CO₂ exchanged between leaves, soil and the atmosphere were well explained by theory describing changes in the oxygen isotope composition of ecosystem water pools in response to changes in leaf transpiration and soil evaporation.     

Nitrogen source and water regime effects on durum wheat photosynthesis and stable carbon and nitrogen isotope composition

Water stress and nitrogen (N) availability are the main constraints limiting yield in durum wheat (Triticum turgidum L. var. durum). This work investigates the combined effects of N source (ammonium–NH₄⁺, nitrate–NO₃^– or a mixture of both–NH₄⁺:NO₃^–) and water availability (well‐watered vs. moderate water stress) on photosynthesis and water‐use efficiency in durum wheat (cv. Korifla) flag leaves grown under controlled conditions, using gas exchange, chlorophyll fluorescence and stable carbon isotope composition (δ¹³C). Under well‐watered conditions, NH₄⁺‐grown plants had lower net assimilation rates (A) than those grown with the other two N forms. This effect was mainly due to lower stomatal conductance (g_s). Under moderate water stress, differences among N forms were not significant, because water regime (WR) had a stronger effect on g_s and A than did N source. Consistent with lower g_s, δ¹³C and transpiration efficiency (TE) were the highest in NH₄⁺ leaves in both water treatments. These results indicate higher water‐use efficiency in plants fertilized with NH₄⁺ due to stomatal limitation on photosynthesis. Moreover, leaf δ¹³C is an adequate trait to assess differences in photosynthetic activity and water‐use efficiency caused by different N sources. Further, the effect of these growing conditions on the nitrogen isotope composition (δ¹⁵N) of flag leaves and roots was examined. Water stress increased leaf δ¹⁵N in all N forms. In addition, leaf δ¹⁵N increased as root N decreased and as leaf δ¹³C became less negative. Regardless of WR, the leaf δ¹⁵N of NO₃^–‐grown plants was lowest. Based on stepwise and canonical discriminant analyses, we conclude that plant δ¹⁵N together with δ¹³C and other variables may reflect the conditions of N nutrition and water availability where the plants were grown. Thus well‐watered plants grown with NH₄⁺:NO₃^– resembled those grown with NO3^–, whereas under water stress they were closer to plants grown with NH₄⁺.     

Quantification of ammonia exchange between agricultural cropland and the atmosphere: Measurements over two complete growth cycles of oilseed rape,wheat, barley and pea

The exchange of ammonia between the atmosphere and the canopy of barley, wheat, oilseed rape and pea crops was studied over two growing seasons by use of a modified aerodynamic gradient technique in which passive horizontal flux samplers were applied with a wind profile in gradient configuration. The crop foliage was a net source of NH₃ to the atmosphere, with NH₃ emissions on a seasonal basis between 1 and 5 kg NH₃–N ha^–1. The amount of NH₃ lost constituted between 1 and 4% of the applied nitrogen and between 1 and 4% of the actual amount of nitrogen present in the mature shoots. The volatile NH₃ losses depended on seasonal variations in climatic conditions affecting the growth and nitrogen economy of the crops and increased under conditions with excessive N absorption by roots and a high N concentration in the foliage. The accumulated NH₃ loss was positively correlated with the above-ground crop N content at anthesis, but not with that at final maturity. There were no indications that NH₃ emissions were larger under conditions unfavourable for nitrogen remobilization from vegetative plant parts (low N harvest index). Nevertheless, a distinct peak in NH₃ emission occurred during senescence. It is concluded that crops in many areas will represent a significant input of ammonia to the atmosphere and that NH₃ losses may become large enough to significantly affect crop N budgets.     

The role of the atmosphere in nitrogen cycling

In this work an analysis of the sources, atmospheric concentration, chemical reactions and sinks of the principal atmospheric nitrogen compounds is made. Atmospheric emissions of N₂O and NH₃ are almost entirely due to biological activity on the continents and in the oceans. The combustion of fossil fuels and biomass is the principal source of NO_x. The only relevant chemical transformations in the troposphere are the oxidation of NO_x to NO₃ ^? and the formation of ammonium salts. Only 10% of the NH₃ emitted is oxidized. Washout of NH₄ ⁺ and NO₃ ^? by rainfall is the principal mechanism for removing nitrogen compounds from the atmosphere. Part of the N₂O enters the stratosphere and part must be removed in the biosphere by processes not yet established. NO_x produced in the atmosphere by the burning of fossil fuels and biomass and by lightning represents between 30 and 40% of the total nitrogen fixed. A complete nitrogen balance for the troposphere is presented. Since the photochemical oxidation of NO_x is rapid and atmospheric transport is relatively slow with respect to the cycling of water in the troposphere, nitrogen compounds return to the earth's surface close to where they were emitted. Fixed-nitrogen inputs to the continents and oceans due to biological and industrial fixation are slightly greater than those due to rain water. However, since rain falls everywhere, input from this source is only important on soils not subject to intensive agriculture.     

Canopy‐scale relationships between stomatal conductance and photosynthesis in irrigated rice

Modeling stomatal behavior is critical in research on land–atmosphere interactions and climate change. The most common model uses an existing relationship between photosynthesis and stomatal conductance. However, its parameters have been determined using infrequent and leaf‐scale gas‐exchange measurements and may not be representative of the whole canopy in time and space. In this study, we used a top‐down approach based on a double‐source canopy model and eddy flux measurements throughout the growing season. Using this approach, we quantified the canopy‐scale relationship between gross photosynthesis and stomatal conductance for 3 years and their relationships with leaf nitrogen content throughout each growing season above a paddy rice canopy in Japan. The canopy‐averaged stomatal conductance (g_sc) increased with increasing gross photosynthesis per unit green leaf area (A_g), as was the case with leaf‐scale measurements, and 41–90% of its variation was explained by variations in A_g adjusted to account for the leaf‐to‐air vapor‐pressure deficit and CO₂ concentration using the Leuning model. The slope (m) in this model (g_sc versus the adjusted A_g) was almost constant within a 15‐day period, but changed seasonally. The m values determined using an ensemble dataset for two mid‐growing‐season 15‐day periods were 30.8 (SE = 0.5), 29.9 (SE = 0.7), and 29.9 (SE = 0.6) in 2004, 2005, and 2006, respectively; the overall mid‐season value was 30.3 and did not greatly differ among the 3 years. However, m appeared to be higher during the early and late growing seasons. The ontogenic changes in leaf nitrogen content strongly affected A_g and thus g_sc. In addition, we have discussed the agronomic impacts of the interactions between leaf nitrogen content and g_sc. Despite limitations in the observations and modeling, our canopy‐scale results emphasize the importance of continuous, season‐long estimates of stomatal model parameters for crops using top‐down approaches.     

Modeling dynamics of stable carbon isotopic exchange between a boreal forest ecosystem and the atmosphere

Stable isotopes of CO₂ contain unique information on the biological and physical processes that exchange CO₂ between terrestrial ecosystems and the atmosphere. In this study, we developed an integrated modeling system to simulate dynamics of stable carbon isotope of CO₂, as well as moisture, energy, and momentum, between a boreal forest ecosystem and the atmosphere, as well as their transport/mixing processes through the convective boundary layer (CBL), using remotely sensed surface parameters to characterize the surface heterogeneity. It has the following characteristics: (i) it accounts for the influences of the CBL turbulent mixing and entrainment of the air aloft; (ii) it scales individual leaf‐level photosynthetic discrimination up to the whole canopy (Δ_canopy) through the separation of sunlit and shaded leaf groups; (iii) it has the capacity to examine the detailed interrelationships among plant water‐use efficiency, isotope discrimination, and vapor pressure deficit; and (iv) it has the potential to investigate how an ecosystem discriminates against ¹³C at various time and spatial scales. The monthly mean isotopic signatures of ecosystem respiration (i.e. δ¹³C_R) used for isotope flux calculation are retrieved from the nighttime flask data from the intensive campaigns (1998–2000) at 20 m level on Fraserdale tower, and the data from the growing season in 1999 are used for model validation. Both the simulated CO₂ mixing ratio and δ¹³C of CO₂ at the 20 m level agreed with the measurements well in different phases of the growing season. On a diurnal basis, the greatest photosynthetic discrimination at canopy level (i.e. Δ_canopy) occurred early morning and late afternoon with a varying range of 10–26‰. The diurnal variability of Δ_canopy was also associated with the phases of growing season and meteorological variables. The annual mean Δ_canopy in 1999 was computed to be 19.58‰. The monthly averages of Δ_canopy varied between 18.55‰ and 20.84‰ with a seasonal peak during the middle growing season. Because of the strong opposing influences of respired and photosynthetic fluxes on forest air (both CO₂ and ¹³CO₂) on both the diurnal and seasonal time scales, CO₂ was consistently enriched with the heavier ¹³C isotope (less negative δ¹³C) from July to October and depleted during the remaining months, whereas on a diurnal basis, CO₂ was enriched with the heavier ¹³C in the late afternoon and depleted in early morning. For the year 1999, the model results reveal that the boreal ecosystem in the vicinity of Fraserdale tower was a small sink with net uptake of 29.07 g ¹²C m^?2 yr^?1 and 0.34 g ¹³C m^?2 yr^?1.     

Climate-ameliorating measures influence photosynthetic gas exchange of apple leaves

Sunburn has become one of the major threats to apple fruit production in South Africa and other countries with Mediterranean climate. Some climate‐ameliorating measures have been developed to control sunburn in apples. Effects of the climate‐ameliorating measures, viz. evaporative cooling, Surround^® WP and shade net, on leaf gas exchange of a 5‐year‐old orchard of ‘Cripps’ Pink’ apple were investigated during hot summer days in Stellenbosch, South Africa. Evaporative cooling increased net photosynthetic rate (A) and stomatal conductance (g_s) because of its lowering of leaf temperature and leaf‐to‐air vapour pressure difference (VPD). Shade net also reduced leaf temperature because of reduction in photosynthetic photon flux density (PPFD). Quantum efficiency of photosynthesis was increased under shade net to compensate for reduced PPFD. Shade net also reduced transpiration rate more than A, resulting in increased midday water‐use efficiency. The diurnal trends of A and g_s in the Surround WP and control treatments were similar, indicating limited ameliorative impact of Surround WP. Furthermore, Surround WP typically reduced maximum rate of carboxylation and the light‐saturated rate of electron transport. In all treatments, A decreased by 70% when leaf temperature increased from 35°C to 40°C. In conclusion, all treatments affected leaf photosynthetic gas exchange. Evaporative cooling enhanced leaf A and g_s because of distinct ameliorative effects on leaf temperature and VPD. Shade net reduced leaf temperature with no consistent effects on leaf gas exchange attributes. Surround WP had limited or no impact on leaf temperature and negatively affected leaf gas exchange attributes.     

The use of stable isotopes to study ecosystem gas exchange

Stable isotopes are a powerful research tool in environmental sciences and their use in ecosystem research is increasing. In this review we introduce and discuss the relevant details underlying the use of carbon and oxygen isotopic compositions in ecosystem gas exchange research. The current use and potential developments of stable isotope measurements together with concentration and flux measurements of CO₂ and water vapor are emphasized. For these applications it is critical to know the isotopic identity of specific ecosystem components such as the isotopic composition of CO₂, organic matter, liquid water, and water vapor, as well as the associated isotopic fractionations, in the soil-plant- atmosphere system. Combining stable isotopes and concentration measurements is very effective through the use of ”Keeling plots.” This approach allows the identification of the isotopic composition and the contribution of ecosystem, or ecosystem components, to the exchange fluxes with the atmosphere. It also allows the estimation of net ecosystem discrimination and soil disequilibrium effects. Recent modifications of the Keeling plot approach permit examination of CO₂ recycling in ecosystems. Combining stable isotopes with dynamic flux measurements requires precision in isotopic sampling and analysis, which is currently at the limit of detection. Combined with the micrometeorological gradient approach (applicable mostly in grasslands and crop fields), stable isotope measurements allow separation of net CO₂ exchange into photosynthetic and soil respiration components, and the evapotranspiration flux into soil evaporation and leaf transpiration. Similar applications in conjunction with eddy correlation techniques (applicable to forests, in addition to grasslands and crop fields) are more demanding, but can potentially be applied in combination with the Keeling plot relationship. The advance and potential in using stable isotope measurements should make their use a standard component in the limited arsenal of ecosystem-scale research tools. Received: 8 July 1999 / Accepted: 10 January 2000     

Exploring cycad foliage as an archive of the isotopic composition of atmospheric nitrogen

Molecular nitrogen (N₂) constitutes the majority of Earth's modern atmosphere, contributing ~0.79 bar of partial pressure (pN₂). However, fluctuations in pN₂ may have occurred on 10⁷–10⁹ year timescales in Earth's past, perhaps altering the isotopic composition of atmospheric nitrogen. Here, we explore an archive that may record the isotopic composition of atmospheric N₂ in deep time: the foliage of cycads. Cycads are ancient gymnosperms that host symbiotic N₂‐fixing cyanobacteria in modified root structures known as coralloid roots. All extant species of cycads are known to host symbionts, suggesting that this N₂‐fixing capacity is perhaps ancestral, reaching back to the early history of cycads in the late Paleozoic. Therefore, if the process of microbial N₂ fixation records the δ¹⁵N value of atmospheric N₂ in cycad foliage, the fossil record of cycads may provide an archive of atmospheric δ¹⁵N values. To explore this potential proxy, we conducted a survey of wild cycads growing in a range of modern environments to determine whether cycad foliage reliably records the isotopic composition of atmospheric N₂. We find that neither biological nor environmental factors significantly influence the δ¹⁵N values of cycad foliage, suggesting that they provide a reasonably robust record of the δ¹⁵N of atmospheric N₂. Application of this proxy to the record of carbonaceous cycad fossils may not only help to constrain changes in atmospheric nitrogen isotope ratios since the late Paleozoic, but also could shed light on the antiquity of the N₂‐fixing symbiosis between cycads and cyanobacteria.     

Evaporation and carbonic anhydrase activity recorded in oxygen isotope signatures of net CO2 fluxes from a Mediterranean soil

The oxygen stable isotope composition (δ¹⁸O) of CO₂ is a valuable tool for studying the gas exchange between terrestrial ecosystems and the atmosphere. In the soil, it records the isotopic signal of water pools subjected to precipitation and evaporation events. The δ¹⁸O of the surface soil net CO₂ flux is dominated by the physical processes of diffusion of CO₂ into and out of the soil and the chemical reactions during CO₂–H₂O equilibration. Catalytic reactions by the enzyme carbonic anhydrase, reducing CO₂ hydration times, have been proposed recently to explain field observations of the δ¹⁸O signatures of net soil CO₂ fluxes. How important these catalytic reactions are for accurately predicting large‐scale biosphere fluxes and partitioning net ecosystem fluxes is currently uncertain because of the lack of field data. In this study, we determined the δ¹⁸O signatures of net soil CO₂ fluxes from soil chamber measurements in a Mediterranean forest. Over the 3 days of measurements, the observed δ¹⁸O signatures of net soil CO₂ fluxes became progressively enriched with a well‐characterized diurnal cycle. Model simulations indicated that the δ¹⁸O signatures recorded the interplay of two effects: (1) progressive enrichment of water in the upper soil by evaporation, and (2) catalytic acceleration of the isotopic exchange between CO₂ and soil water, amplifying the contributions of ‘atmospheric invasion’ to net signatures. We conclude that there is a need for better understanding of the role of enzymatic reactions, and hence soil biology, in determining the contributions of soil fluxes to oxygen isotope signals in atmospheric CO₂.     

Leaf Gas Exchange of Pachyrhizus ahipa and P. erosus Under Water and Temperature Stress

Gas exchange, water relations, and leaf traits were studied in the tuberous-root producing legumes ahipa (Pachyrhizus ahipa) and yambean (P. erosus) under different environmental conditions. Differences in leaf traits (hairiness, leaf area, areal leaf mass, stomatal density) and paraheliotropism were found between ahipa and yambean. Under sufficient water supply, the increase in air temperature and decrease in air humidity increased stomatal conductance (g _s) and net photosynthetic rate (P _N) in yambean but reduced them in ahipa. In a drying soil (14 d after irrigation), inter-specific variation in gas exchange was only observed in the early morning, and yambean showed a greater sensitivity to water restriction than ahipa. High g _s at low humidity increased P _N of P. erosus but resulted in lower water-use efficiency (WUE). However, long-term WUE, estimated by leaf carbon isotope discrimination, showed little variation between species. Daily-irrigated ahipa and yambean grown in the greenhouse did not show significant differences in gas exchange. However, leaf temperature was significantly greater in yambean than in ahipa while a steepper relationship between E and P _N and g _s was observed in ahipa.     

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

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

Informing climate models with rapid chamber measurements of forest carbon uptake

Models predicting ecosystem carbon dioxide (CO₂) exchange under future climate change rely on relatively few real‐world tests of their assumptions and outputs. Here, we demonstrate a rapid and cost‐effective method to estimate CO₂ exchange from intact vegetation patches under varying atmospheric CO₂ concentrations_. We find that net ecosystem CO₂ uptake (NEE) in a boreal forest rose linearly by 4.7 ± 0.2% of the current ambient rate for every 10 ppm CO₂ increase, with no detectable influence of foliar biomass, season, or nitrogen (N) fertilization. The lack of any clear short‐term NEE response to fertilization in such an N‐limited system is inconsistent with the instantaneous downregulation of photosynthesis formalized in many global models. Incorporating an alternative mechanism with considerable empirical support – diversion of excess carbon to storage compounds – into an existing earth system model brings the model output into closer agreement with our field measurements. A global simulation incorporating this modified model reduces a long‐standing mismatch between the modeled and observed seasonal amplitude of atmospheric CO₂. Wider application of this chamber approach would provide critical data needed to further improve modeled projections of biosphere–atmosphere CO₂ exchange in a changing climate.     

Nitrogen accumulation and distribution in Danthonia richardsonii swards in response to CO2 and nitrogen supply over four years of growth

Nitrogen‐stressed microcosms of the C3 grass Danthonia richardsonii gained nitrogen from the environment when grown under ambient or enriched (359, ‘amb’ or 719 μL L^{? 1}‘enr’, respectively) atmospheric CO₂ concentrations over a 4‐y period. This gain was apparent at all rates of supplied mineral N (2.2, 6.7 or 19.8 g N m^{? 2} y^{? 1}– low‐N, mid‐N or high‐N), although it was small at high‐N. Small losses of N occurred from the microcosm as leachate, while gaseous losses of N were estimated to be between 10% and 25% of applied mineral N. Losses of applied mineral N were slightly lower under CO₂ enrichment only at the highest rate of mineral N supply. Levels of ¹⁵N natural abundance in green leaf (δ¹⁵Ν) of ? 2‰ (amb low‐N) and of below ? 4‰ (enr low‐ & mid‐N) suggest that absorption of atmospheric NH₃ may have been a source of some of the extra N in the low and mid‐N treatments. Biological N₂ fixation, of up to 2 g m^{? 2} y^{? 1} was hypothesized to form the remainder of the environmental N source. Microcosm C:N ratio was higher under CO₂ enrichment. Nitrogen productivity of microcosm carbon gain (g C accumulated g^{? 1} leaf N day^{? 1}) was increased (up to 100%) by CO₂ enrichment at all rates of mineral N supply. Green leaf %N was reduced by CO₂ enrichment, and there was less nitrogen in the green leaf pool under CO₂ enrichment. Less, or the same amount of nitrogen was present in senesced leaf, surface litter and root under CO₂ enrichment while more nitrogen was present in the soil in organic forms, and as NH₄ + at the highest rate of mineral N supply.     

Isotopic evidence for the occurrence of biological nitrification and nitrogen deposition processing in forest canopies

This study examines the role of tree canopies in processing atmospheric nitrogen (N_dep) for four forests in the United Kingdom subjected to different N_dep: Scots pine and beech stands under high N_dep (HN, 13–19 kg N ha^?1 yr^?1), compared to Scots pine and beech stands under low N_dep (LN, 9 kg N ha^?1 yr^?1). Changes of NO₃‐N and NH₄‐N concentrations in rainfall (RF) and throughfall (TF) together with a quadruple isotope approach, which combines δ¹⁸O, Δ¹⁷O and δ¹⁵N in NO₃^? and δ¹⁵N in NH₄⁺, were used to assess N transformations by the canopies. Generally, HN sites showed higher NH₄‐N and NO₃‐N concentrations in RF compared to the LN sites. Similar values of δ¹⁵N‐NO₃^? and δ¹⁸O in RF suggested similar source of atmospheric NO₃^? (i.e. local traffic), while more positive values for δ¹⁵N‐NH₄⁺ at HN compared to LN likely reflected the contribution of dry NH_x deposition from intensive local farming. The isotopic signatures of the N‐forms changed after interacting with tree canopies. Indeed, ¹⁵N‐enriched NH₄⁺ in TF compared to RF at all sites suggested that canopies played an important role in buffering dry N_dep also at the low N_dep site. Using two independent methods, based on δ¹⁸O and Δ¹⁷O, we quantified for the first time the proportion of NO₃^? in TF, which derived from nitrification occurring in tree canopies at the HN site. Specifically, for Scots pine, all the considered isotope approaches detected biological nitrification. By contrast for the beech, only using the mixing model with Δ¹⁷O, we were able to depict the occurrence of nitrification within canopies. Our study suggests that tree canopies play an active role in the N cycling within forest ecosystems. Processing of N_dep within canopies should not be neglected and needs further exploration, with the combination of multiple isotope tracers, with particular reference to Δ¹⁷O.