Nitrogen deposition outweighs climatic variability in driving annual growth rate of canopy beech trees: Evidence from long‐term growth reconstruction across a geographic gradient

In this study, we investigated the role of climatic variability and atmospheric nitrogen deposition in driving long‐term tree growth in canopy beech trees along a geographic gradient in the montane belt of the Italian peninsula, from the Alps to the southern Apennines. We sampled dominant trees at different developmental stages (from young to mature tree cohorts, with tree ages spanning from 35 to 160 years) and used stem analysis to infer historic reconstruction of tree volume and dominant height. Annual growth volume (G_V) and height (G_H) variability were related to annual variability in model simulated atmospheric nitrogen deposition and site‐specific climatic variables, (i.e. mean annual temperature, total annual precipitation, mean growing period temperature, total growing period precipitation, and standard precipitation evapotranspiration index) and atmospheric CO₂ concentration, including tree cambial age among growth predictors. Generalized additive models (GAM), linear mixed‐effects models (LMM), and Bayesian regression models (BRM) were independently employed to assess explanatory variables. The main results from our study were as follows: (i) tree age was the main explanatory variable for long‐term growth variability; (ii) GAM, LMM, and BRM results consistently indicated climatic variables and CO₂ effects on G_V and G_H were weak, therefore evidence of recent climatic variability influence on beech annual growth rates was limited in the montane belt of the Italian peninsula; (iii) instead, significant positive nitrogen deposition (N_dep) effects were repeatedly observed in G_V and G_H; the positive effects of N_dep on canopy height growth rates, which tended to level off at N_dep values greater than approximately 1.0 g m^?2 y^?1, were interpreted as positive impacts on forest stand above‐ground net productivity at the selected study sites.     

Last‐century changes of alpine grassland water‐use efficiency: a reconstruction through carbon isotope analysis of a time‐series of Capra ibex horns

The ecophysiological response of an alpine grassland to recent climate change and increasing atmospheric CO₂ concentration was investigated with a new strategy to go back in time: using a time‐series of Capra ibex horns as archives of the alpine grasslands' carbon isotope discrimination (¹³Δ). From the collection of the Natural History Museum of Bern, horns of 24 males from the population of the Augstmatthorn–Brienzer Rothorn mountains, Switzerland, were sampled covering the period from 1938 to 2006. Samples were taken from the beginning of each year‐ring of the horns, representing the beginning of the horn growth period, the spring. The horns' carbon ¹³C content (Δ¹³C) declined together with that of atmospheric CO₂ over the 69‐year period, but ¹³Δ increased slightly (+0.4‰), though significantly (P<0.05), over the observation period. Estimated intercellular CO₂ concentration increased (+56 μmol mol^?1) less than the atmospheric CO₂ concentration (+81 μmol mol^?1), so that intrinsic water‐use efficiency increased by 17.8% during the 69‐year period. However, the atmospheric evaporative demand at the site increased by approximately 0.1 kPa between 1955 and 2006, thus counteracting the improvement of intrinsic water‐use efficiency. As a result, instantaneous water‐use efficiency did not change. The observed changes in intrinsic water‐use efficiency were in the same range as those of trees (as reported by others), indicating that leaf‐level control of water‐use efficiency of grassland and forests followed the same principles. This is the first reconstruction of the water‐use efficiency response of a natural grassland ecosystem to last century CO₂ and climatic changes. The results indicate that the alpine grassland community has responded to climate change by improving the physiological control of carbon gain to water loss, following the increases in atmospheric CO₂ and evaporative demand. But, effective leaf‐level water‐use efficiency has remained unchanged.     

Nitrogen deposition and plant species interact to influence soil carbon stabilization

Anthropogenic nitrogen (N) deposition effects on soil organic carbon (C) decomposition remain controversial, while the role of plant species composition in mediating effects of N deposition on soil organic C decomposition and long‐term soil C sequestration is virtually unknown. Here we provide evidence from a 5‐year grassland field experiment in Minnesota that under elevated atmospheric CO₂ concentration (560 ppm), plant species determine whether N deposition inhibits the decomposition of soil organic matter via inter‐specific variation in root lignin concentration. Plant species producing lignin‐rich litter increased stabilization of soil C older than 5 years, but only in combination with elevated N inputs (4 g m^?2 year^?1). Our results suggest that N deposition will increase soil C sequestration in those ecosystems where vegetation composition and/or elevated atmospheric CO₂ cause high litter lignin inputs to soils.     

Vertical canopy gradients in δ13C correspond with leaf nitrogen content in a mixed-species conifer forest

Stable carbon isotope composition varies markedly between sun and shade leaves, with sun leaves being invariably more enriched (i.e., they contain more¹³C). Several hypotheses have emerged to explain this pattern, but controversy remains as to which mechanism is most general. We measured vertical gradients in stable carbon isotope composition (δ¹³C) in more than 200 trees of nine conifer species growing in mixed-species forests in the Northern Rocky Mountains, USA. For all species except western larch, δ¹³C decreased from top to bottom of the canopy. We found that δ¹³C was strongly correlated with nitrogen per unit leaf area (N _area), which is a measure of photosynthetic capacity. Usually weaker correlations were found between δ¹³C and leaf mass per area, nitrogen per unit leaf mass, height from the ground, or depth in the canopy, and these correlations were more variable between trees than for N _area. Gradients of δ¹³C (per meter canopy depth) were steeper in small trees than in tall trees, indicating that a recent explanation of δ¹³C gradients in terms of drought stress of upper canopy leaves is unlikely to apply in our study area. The strong relationship between N _area and δ¹³C here reported is consistent with the general finding that leaves or species with higher photosynthetic capacity tend to maintain lower CO₂ concentrations inside leaves. We conclude that photosynthetic capacity is a strong determinant of δ¹³C in vertical canopy profiles, and must be accounted for when interpreting δ¹³C values in conifer forests.     

Elevated atmospheric CO2 and increased nitrogen deposition: effects on C and N metabolism and growth of the peat moss Sphagnum recurvum P. Beauv. var. mucronatum (Russ.) Warnst

Sphagnum bogs play an important role when considering the impacts of global change on global carbon and nitrogen cycles. Sphagnum recurvum P. Beauv. var. mucronatum (Russ.) was grown at 360 (ambient) and 700 μL L^?1 (elevated) atmospheric [CO₂] in combination with different nitrogen deposition rates (6, 15, 23 g N m^?2 y^?1), in a short‐ and long‐term growth chamber experiment. After 6 months, elevated atmospheric [CO₂] in combination with the lowest nitrogen deposition rate, increased plant dry mass by 17%. In combination with a high nitrogen deposition rate, biomass production was not significantly stimulated. At the start of the experiment, photosynthesis was stimulated by elevated atmospheric [CO₂], but it was downregulated to control levels after three days of exposure. Elevated [CO₂] substantially reduced dark respiration, which resulted in a continuous increase in soluble sugar content in capitula. Differences in growth response among different nitrogen and CO₂ treatments could not be related to measured carbon exchange rates, which was mainly due to interference of microbial respiration. Doubling atmospheric [CO₂] reduced total nitrogen content in capitula but not in stems at all nitrogen deposition rates. Reduction in total nitrogen content coincided with a decrease in amino acids, but soluble protein levels remained unaffected. Thus, elevated [CO₂] induced a substantial shift in the partitioning of nitrogen compounds in capitula. Soluble sugar concentration was negatively correlated with total nitrogen content, which implies that the reduction in amino acid content in capitula, exposed to elevated [CO₂], might be caused by the accumulation of soluble sugars. Growth was not stimulated by increased nitrogen deposition. High nitrogen deposition, resulting in a capitulum nitrogen content in excess of 15 mg g^?1 dw, was detrimental to photosynthesis, reduced water content and induced necrosis. We propose a capitulum nitrogen content of 15 mg g^?1 dw as a possible bioindicator for the detection of nitrogen pollution stress in oligo‐mesotrophic peat bog ecosystems. At the lowest nitrogen deposition level, nitrogen recovery was higher than 100%, which indicates substantial dry deposition and/or gaseous nitrogen fixation by bacteria, associated with Sphagnum. Increasing nitrogen deposition rates decreased nitrogen recovery percentages, which indicates reduced efficiency of nitrogen fixation.     

The effects of nitrogen stress on the stable carbon isotope composition, productivity and water use efficiency of white spruce (Picea glauca (Moench) Voss) seedlings

A, assimilation rate
a, fractionation against ¹³C for CO₂ diffusion through air
b, net fractionation against ¹³C during CO₂ fixation
C_a, ambient CO₂ concentration
C_c, CO₂ concentration at the chloroplast
C_i, intercellular CO₂ concentration
D, vapour pressure deficit
E_n, needle transpiration rate
E_p, whole plant water use
g_w, leaf internal transfer conductance to CO₂
g_s, stomatal conductance to water vapour
L, projected leaf area
NUE, nitrogen use efficiency
PEP, phosphoenolpyruvate
Rubisco, ribulose-1,5-biphosphate carboxylase
TDR, time domain reflectometry
WUE, water use efficiency
Δ, carbon isotope discrimination
δ¹³C, carbon isotope abundance parameter
δ¹³C_a, carbon isotopic composition of atmospheric CO₂
θ, volumetric soil water content

The effect of nitrogen stress on needle δ¹³C, water-use efficiency (WUE) and biomass production in irrigated and dry land white spruce (Picea glauca (Moench) Voss) seedlings was investigated. Sixteen hundred seedlings, representing 10 controlled crosses, were planted in the field in individual buried sand-filled cylinders. Two nitrogen treatments were imposed, nitrogen stressed and fertilized. The ranking of δ¹³C of the crosses was maintained across all combinations of water and nitrogen treatments and there was not a significant genetic versus environmental interaction. The positive relationships between needle δ¹³C, WUE and dry matter production demonstrate that it should be possible to use δ¹³C as a surrogate for WUE, and to select for increased WUE without compromising yield, even in nitrogen deficient environments. Nitrogen stressed seedlings had the lowest needle δ¹³C in both irrigated and dry land conditions. There was a positive correlation between needle nitrogen content and δ¹³C that was likely associated with increased photosynthetic capacity. There was some indication that decreased nitrogen supply led to increased stomatal conductance and hence lower WUE. There was a negative correlation between intrinsic water use efficiency and photosynthetic nitrogen use efficiency (NUE). This suggests that white spruce seedlings have the ability to maximize NUE when water becomes limited. There was significant genetic variation in NUE that was maintained across treatments. Our results suggest that in white spruce, there is no detectable effect of anaplerotic carbon fixation and that it is more appropriate to use a value of 29‰ (‘Rubisco only’) for the net discrimination against ¹³C during CO₂ fixation. This leads to excellent correspondence between values of C_i/C_a derived from gas exchange measurements or from δ¹³C.     

Microbial 13C utilization patterns via stable isotope probing of phospholipid biomarkers in Mojave Desert soils exposed to ambient and elevated atmospheric CO2

Changes in plant inputs under changing atmospheric CO₂ can be expected to alter the size and/or functional characteristics of soil microbial communities which can determine whether soils are a C sink or source. Stable isotope probing was used to trace autotrophically fixed ¹³C into phospholipid fatty acid (PLFA) biomarkers in Mojave Desert soils planted with the desert shrub, Larrea tridentata. Seedlings were pulse‐labeled with ¹³CO₂ under ambient and elevated CO₂ in controlled environmental growth chambers. The label was chased into the soil by extracting soil PLFAs after labeling at Days 0, 2, 10, 24, and 49. Eighteen of 29 PLFAs identified showed ¹³C enrichment relative to nonlabeled control soils. Patterns of PLFA enrichment varied temporally and were similar for various PLFAs found within a microbial functional group. Enrichment of PLFA ¹³C generally occurred within the first 2 days in general and fungal biomarkers, followed by increasingly greater enrichment in bacterial biomarkers as the study progressed (Gram‐negative, Gram‐positive, actinobacteria). While treatment CO₂ level did not affect total PLFA‐C concentrations, microbial functional group abundances and distribution responded to treatment CO₂ level and these shifts persisted throughout the study. Specifically, ratios of bacterial‐to‐total PLFA‐C decreased and fungal‐to‐bacterial PLFA‐C increased under elevated CO₂ compared with ambient conditions. Differences in the timing of ¹³C incorporation into lipid biomarkers coupled with changes in microbial functional groups indicate that microbial community characteristics in Mojave Desert soils have shifted in response to long‐term exposure to increased atmospheric CO₂.     

Variation in the δ13C of foliage of Pinus sylvestris L. in relation to climate and additions of nitrogen: analysis of a 32-year chronology

We report an analysis of both the long‐ and short‐term drivers of the carbon (C) isotope composition (δ¹³C) values of current year needles of Pinus sylvestris L. linked to changing atmospheric carbon dioxide (CO₂) concentrations (c_a) and climate using data from a uniquely long‐term nitrogen (N) fertilization experiment in the north of Sweden (consisting of three N dosage levels and a control treatment) from 1970 until 2002. N loading produced trees with less negative δ¹³C of foliage, by around 0.45‰ on average, with the difference in δ¹³C between control and N treatments not dependant upon N dosage. The average δ¹³C values decreased at a rate of around 0.03‰ yr⁻¹, even after accounting for the Suess effect (the decrease in the atmospheric CO₂δ¹³C due to anthropogenic emissions of isotopically light CO₂). This decrease is large enough to cause a significant, progressive change in the δ¹³C down through a soil profile. Modelled values of plant intrinsic water use efficiency (WUE_i) and the ratio of leaf internal to external [CO₂] (c_i/c_a) showed that this was the result of c_i increasing in parallel with c_a (while c_i/c_a increased), thus causing little change in WUE_i over the 32 years of study. The residuals from the relationships between year and δ¹³C were used to examine the impact of climate on the interannual variation of C isotope composition of needles. This included the use of a fire hazard index (FHI) model, which integrates climatic factors known to influence plant stomatal conductance and hence δ¹³C. The FHI produced the best fit with δ¹³C values when climate data were averaged over the whole growth season (for control plots) and for July for all the N treatments, explaining ca. 60% of the total interannual variation in δ¹³C. Further, trees from the N treatments appeared more susceptible to air‐humidity‐based climate parameters, as seen from higher correlation coefficients, than were control trees. Thus, our data suggest the possibility of increased susceptibility to drought conditions in ecosystems with moderate to high N deposition rates. Also, there is the possibility that, because there was no apparent change in WUE_i of P. sylvestris in this ecosystem over the last 32 years, the rate of sequestration of C into boreal ecosystems may not increase with c_a, as has been predicted.     

Unique Responsiveness of Angiosperm Stomata to Elevated CO2 Explained by Calcium Signalling

Angiosperm and conifer tree species respond differently when exposed to elevated CO_2, with angiosperms found to dynamically reduce water loss while conifers appear insensitive. Such distinct responses are likely to affect competition between these tree groups as atmospheric CO₂ concentration rises. Seeking the mechanism behind this globally important phenomenon we targeted the Ca²⁺-dependent signalling pathway, a mediator of stomatal closure in response to elevated CO₂, as a possible explanation for the differentiation of stomatal behaviours. Sampling across the diversity of vascular plants including lycophytes, ferns, gymnosperms and angiosperms we show that only angiosperms possess the stomatal behaviour and prerequisite genetic coding, linked to Ca²⁺-dependent stomatal signalling. We conclude that the evolution of Ca²⁺-dependent stomatal signalling gives angiosperms adaptive benefits in terms of highly efficient water use, but that stomatal sensitivity to high CO₂ may penalise angiosperm productivity relative to other plant groups in the current era of soaring atmospheric CO₂.     

Impacts of 3 years of elevated atmospheric CO2 on rhizosphere carbon flow and microbial community dynamics

Carbon (C) uptake by terrestrial ecosystems represents an important option for partially mitigating anthropogenic CO₂ emissions. Short‐term atmospheric elevated CO₂ exposure has been shown to create major shifts in C flow routes and diversity of the active soil‐borne microbial community. Long‐term increases in CO₂ have been hypothesized to have subtle effects due to the potential adaptation of soil microorganism to the increased flow of organic C. Here, we studied the effects of prolonged elevated atmospheric CO₂ exposure on microbial C flow and microbial communities in the rhizosphere. Carex arenaria (a nonmycorrhizal plant species) and Festuca rubra (a mycorrhizal plant species) were grown at defined atmospheric conditions differing in CO₂ concentration (350 and 700 ppm) for 3 years. During this period, C flow was assessed repeatedly (after 6 months, 1, 2, and 3 years) by ¹³C pulse‐chase experiments, and label was tracked through the rhizosphere bacterial, general fungal, and arbuscular mycorrhizal fungal (AMF) communities. Fatty acid biomarker analyses and RNA‐stable isotope probing (RNA‐SIP), in combination with real‐time PCR and PCR‐DGGE, were used to examine microbial community dynamics and abundance. Throughout the experiment the influence of elevated CO₂ was highly plant dependent, with the mycorrhizal plant exerting a greater influence on both bacterial and fungal communities. Biomarker data confirmed that rhizodeposited C was first processed by AMF and subsequently transferred to bacterial and fungal communities in the rhizosphere soil. Over the course of 3 years, elevated CO₂ caused a continuous increase in the ¹³C enrichment retained in AMF and an increasing delay in the transfer of C to the bacterial community. These results show that, not only do elevated atmospheric CO₂ conditions induce changes in rhizosphere C flow and dynamics but also continue to develop over multiple seasons, thereby affecting terrestrial ecosystems C utilization processes.     

Enhanced CO2 uptake is marginally offset by altered fluxes of non-CO2 greenhouse gases in global forests and grasslands under N deposition

Despite the increasing impact of atmospheric nitrogen (N) deposition on terrestrial greenhouse gas (GHG) budget, through driving both the net atmospheric CO₂ exchange and the emission or uptake of non-CO₂ GHGs (CH₄ and N₂O), few studies have assessed the climatic impact of forests and grasslands under N deposition globally based on different bottom-up approaches. Here, we quantify the effects of N deposition on biomass C increment, soil organic C (SOC), CH₄ and N₂O fluxes and, ultimately, the net ecosystem GHG balance of forests and grasslands using a global comprehensive dataset. We showed that N addition significantly increased plant C uptake (net primary production) in forests and grasslands, to a larger extent for the aboveground C (aboveground net primary production), whereas it only caused a small or insignificant enhancement of SOC pool in both upland systems. Nitrogen addition had no significant effect on soil heterotrophic respiration (R_H) in both forests and grasslands, while a significant N-induced increase in soil CO₂ fluxes (R_S, soil respiration) was observed in grasslands. Nitrogen addition significantly stimulated soil N₂O fluxes in forests (76%), to a larger extent in grasslands (87%), but showed a consistent trend to decrease soil uptake of CH₄, suggesting a declined sink capacity of forests and grasslands for atmospheric CH₄ under N enrichment. Overall, the net GHG balance estimated by the net ecosystem production-based method (forest, 1.28 Pg CO₂-eq year⁻¹ vs. grassland, 0.58 Pg CO₂-eq year⁻¹) was greater than those estimated using the SOC-based method (forest, 0.32 Pg CO₂-eq year⁻¹ vs. grassland, 0.18 Pg CO₂-eq year⁻¹) caused by N addition. Our findings revealed that the enhanced soil C sequestration by N addition in global forests and grasslands could be only marginally offset (1.5%–4.8%) by the combined effects of its stimulation of N₂O emissions together with the reduced soil uptake of CH₄.     

Nutrient supply enhanced the increase in intrinsic water‐use efficiency of a temperate seminatural grassland in the last century

Under the increase in atmospheric CO₂ during the last century, variable increases in the intrinsic water‐use efficiency (W_i), i.e., the ratio between carbon assimilation rate (A) and stomatal conductance (g_s), of C₃ vegetation have been observed. Here, we ask if long‐term nutrient status and especially nitrogen supply have an effect on the CO₂ response of W_i in a temperate seminatural C₃ grassland. This analysis draws on the long‐term trends (1915–2009) in W_i, derived from carbon isotope analysis, of archived hay and herbage from the Park Grass Experiment at Rothamsted (South‐East England). Plant samples came from five fertilizer treatments, each with different annual nitrogen (N; 0, 48 or 96 kg ha^?1), phosphorus (P; 0 or 35 kg ha^?1) and potassium (K; 0 or 225 kg ha^?1) applications, with lime as required to maintain soil pH near 7. Carbon isotope discrimination (¹³Δ) increased significantly (P < 0.001) on the Control (0.9‰ per 100 ppm CO₂ increase). This trend differed significantly (P < 0.01) from those observed on the fertilized treatments (PK only: 0.4‰ per 100 ppm CO₂ increase, P < 0.001; Low N only, Low N+PK, High N+PK: no significant increase). The ¹³Δ trends on fertilized treatments did not differ significantly from each other. However, N status, assessed as N fertilizer supply plus an estimate of biologically fixed N, was negatively related (r² = 0.88; P < 0.02) to the trend for ¹³Δ against CO₂. Other indices of N status exhibited similar relationships. Accordingly, the increase in W_i at High N+PK was twice that of the Control (+28% resp. +13% relative to 1915). In addition, the CO₂ responsiveness of ¹³Δ was related to the grass content of the plant community. This may have been due to the greater CO₂ responsiveness of g_s in grasses relative to forbs. Thus, the greater CO₂ response of grass‐rich fertilized swards may be related to effects of nutrient supply on botanical composition.     

Soil carbon and nitrogen cycling and storage throughout the soil profile in a sweetgum plantation after 11 years of CO2‐enrichment

Increased partitioning of carbon (C) to fine roots under elevated [CO₂], especially deep in the soil profile, could alter soil C and nitrogen (N) cycling in forests. After more than 11 years of free‐air CO₂ enrichment in a Liquidambar styraciflua L. (sweetgum) plantation in Oak Ridge, TN, USA, greater inputs of fine roots resulted in the incorporation of new C (i.e., C with a depleted δ¹³C) into root‐derived particulate organic matter (POM) pools to 90‐cm depth. Even though production in the sweetgum stand was limited by soil N availability, soil C and N contents were greater throughout the soil profile under elevated [CO₂] at the conclusion of the experiment. Greater C inputs from fine‐root detritus under elevated [CO₂] did not result in increased net N immobilization or C mineralization rates in long‐term laboratory incubations, possibly because microbial biomass was lower in the CO₂‐enriched plots. Furthermore, the δ¹³CO₂ of the C mineralized from the incubated soil closely tracked the δ¹³C of the labile POM pool in the elevated [CO₂] treatment, especially in shallower soil, and did not indicate significant priming of the decomposition of pre‐experiment soil organic matter (SOM). Although potential C mineralization rates were positively and linearly related to total SOM C content in the top 30 cm of soil, this relationship did not hold in deeper soil. Taken together with an increased mean residence time of C in deeper soil pools, these findings indicate that C inputs from relatively deep roots under elevated [CO₂] may increase the potential for long‐term soil C storage. However, C in deeper soil is likely to take many years to accrue to a significant fraction of total soil C given relatively smaller root inputs at depth. Expanded representation of biogeochemical cycling throughout the soil profile may improve model projections of future forest responses to rising atmospheric [CO₂].     

Anthropogenic nitrogen deposition enhances carbon sequestration in boreal soils

It is proposed that carbon (C) sequestration in response to reactive nitrogen (N_r) deposition in boreal forests accounts for a large portion of the terrestrial sink for anthropogenic CO₂ emissions. While studies have helped clarify the magnitude by which N_r deposition enhances C sequestration by forest vegetation, there remains a paucity of long‐term experimental studies evaluating how soil C pools respond. We conducted a long‐term experiment, maintained since 1996, consisting of three N addition levels (0, 12.5, and 50 kg N ha^?1 yr^?1) in the boreal zone of northern Sweden to understand how atmospheric N_r deposition affects soil C accumulation, soil microbial communities, and soil respiration. We hypothesized that soil C sequestration will increase, and soil microbial biomass and soil respiration will decrease, with disproportionately large changes expected compared to low levels of N addition. Our data showed that the low N addition treatment caused a non‐significant increase in the organic horizon C pool of ~15% and a significant increase of ~30% in response to the high N treatment relative to the control. The relationship between C sequestration and N addition in the organic horizon was linear, with a slope of 10 kg C kg^?1 N. We also found a concomitant decrease in total microbial and fungal biomasses and a ~11% reduction in soil respiration in response to the high N treatment. Our data complement previous data from the same study system describing aboveground C sequestration, indicating a total ecosystem sequestration rate of 26 kg C kg^?1 N. These estimates are far lower than suggested by some previous modeling studies, and thus will help improve and validate current modeling efforts aimed at separating the effect of multiple global change factors on the C balance of the boreal region.     

Effect of climate change,CO2 trends,nitrogen addition,and land‐cover and management intensity changes on the carbon balance of European grasslands

Several lines of evidence point to European managed grassland ecosystems being a sink of carbon. In this study, we apply ORCHIDEE‐GM a process‐based carbon cycle model that describes specific management practices of pastures and the dynamics of carbon cycling in response to changes in climatic and biogeochemical drivers. The model is used to simulate changes in the carbon balance [i.e., net biome production (NBP)] of European grasslands over 1991–2010 on a 25 km × 25 km grid. The modeled average trend in NBP is 1.8–2.0 g C m^?2 yr^?2 during the past two decades. Attribution of this trend suggests management intensity as the dominant driver explaining NBP trends in the model (36–43% of the trend due to all drivers). A major change in grassland management intensity has occurred across Europe resulting from reduced livestock numbers. This change has ‘inadvertently’ enhanced soil C sequestration and reduced N₂O and CH₄ emissions by 1.2–1.5 Gt CO₂‐equivalent, offsetting more than 7% of greenhouse gas emissions in the whole European agricultural sector during the period 1991–2010. Land‐cover change, climate change and rising CO₂ also make positive and moderate contributions to the NBP trend (between 24% and 31% of the trend due to all drivers). Changes in nitrogen addition (including fertilization and atmospheric deposition) are found to have only marginal net effect on NBP trends. However, this may not reflect reality because our model has only a very simple parameterization of nitrogen effects on photosynthesis. The sum of NBP trends from each driver is larger than the trend obtained when all drivers are varied together, leaving a residual – nonattributed – term (22–26% of the trend due to all drivers) indicating negative interactions between drivers.     

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.     

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

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

Plasticity in photosynthetic response to nutrient supply of seedlings from a mixed conifer‐angiosperm forest

Abstract We measured the plasticity of the response of photosynthesis to nutrient supply in seedlings of the dominant four conifer and broadleaved angiosperm tree species from an indigenous forest in South‐westland, New Zealand. We hypothesized that the response of conifers to differing nutrient supply would be less than the response for the angiosperms because of greater adaptation to low fertility conditions. In Prumnopitys ferruginea (D. Don) de Laub. the maximum velocity of electron transport, J_max, doubled with a 10‐fold increase in concentration of nitrogen supply. In Dacrydium cupressinum Lamb. the maximum velocity of carboxylation, V_cmax, doubled with a 10‐fold increase in phosphorus supply. In contrast, photosynthetic capacity for the angiosperm species Weinmannia racemosa L.f. was affected only by the interaction of nitrogen and phosphorus and photosynthetic capacity of Metrosideros umbellata Cav. was not affected by nutrient supply. The response of the conifers to increasing availability of nutrient suggests greater plasticity in photosynthetic capacity, a characteristic not generally associated with adaptation to soil infertility, thus invalidating our hypothesis. Our data suggest that photosynthetic response to nutrient supply cannot be broadly generalized between the two functional groups.     

Temperature‐induced water stress in high‐latitude forests in response to natural and anthropogenic warming

The Arctic is particularly sensitive to climate change, but the independent effects of increasing atmospheric CO₂ concentration (pCO₂) and temperature on high‐latitude forests are poorly understood. Here, we present a new, annually resolved record of stable carbon isotope (δ¹³C) data determined from Larix cajanderi tree cores collected from far northeastern Siberia in order to investigate the physiological response of these trees to regional warming. The tree‐ring record, which extends from 1912 through 1961 (50 years), targets early twentieth‐century warming (ETCW), a natural warming event in the 1920s to 1940s that was limited to Northern hemisphere high latitudes. Our data show that net carbon isotope fractionation (Δ¹³C), decreased by 1.7‰ across the ETCW, which is consistent with increased water stress in response to climate warming and dryer soils. To investigate whether this signal is present across the northern boreal forest, we compiled published carbon isotope data from 14 high‐latitude sites within Europe, Asia, and North America. The resulting dataset covered the entire twentieth century and spanned both natural ETCW and anthropogenic Late Twentieth‐Century Warming (~0.7 °C per decade). After correcting for a ~1‰ increase in Δ¹³C in response to twentieth century pCO₂ rise, a significant negative relationship (r = ?0.53, P < 0.0001) between the average, annual Δ¹³C values and regional annual temperature anomalies is observed, suggesting a strong control of temperature on the Δ¹³C value of trees growing at high latitudes. We calculate a 17% increase in intrinsic water‐use efficiency within these forests across the twentieth century, of which approximately half is attributed to a decrease in stomatal conductance in order to conserve water in response to drying conditions, with the other half being attributed to increasing pCO₂. We conclude that annual tree‐ring records from northern high‐latitude forests record the effects of climate warming and pCO₂ rise across the twentieth century.     

The combined effect of constant water deficit and nitrogen supply on WUE, NUE and Δ13C in durum wheat potted plants

Water scarcity and nitrogen shortage are the main constraints on durum wheat productivity. This paper examines the combined effects of a constant water deficit and nitrogen supply (NS) on growth, photosynthesis, stomatal conductance (g_s) and transpiration, instantaneous and time‐integrated water use efficiency (WUE) and nitrogen use efficiency (NUE) and carbon isotope discrimination (Δ¹³C) in durum wheat genotypes grown in pots under greenhouse conditions. Three water levels (40%, 70% and 100% container capacity), two nitrogen doses (high and low N) and four genotypes were assayed in a total of 24 experimental treatments. Water and nitrogen treatments were imposed 2 weeks after plant emergence. The growth, nitrogen content and Δ¹³C of the shoot and the gas exchange in the flag leaf were determined about 2 weeks after anthesis. As expected, both water and NS had a strong positive effect on growth. However, a reduction in water supply had low effect decreasing photosynthesis and transpiration, Δ¹³C and NUE and increasing WUE. On the contrary, increasing the level of nitrogen supplied had a significant negative effect on g_s, which decreased significantly the ratio of intercellular to ambient CO₂ concentrations and Δ¹³C, and increased both instantaneous and time‐integrated WUE. In addition, a higher N level also negatively affected the instantaneous and time‐integrated NUE. The Δ¹³C of shoots correlated significantly and negatively with either instantaneous or time‐integrated measurements of WUE. Moreover, within each NS, Δ¹³C also correlated negatively with the integrated NUE. We concluded that under our experimental conditions, Δ¹³C gives information about the efficiency with which not just water but also nitrogen are used by the plant. In addition, this study illustrates that a steady water limitation may strongly affect biomass without consistent changes in WUE. The lack of effect of the different water regimes on gas exchange, WUE and Δ¹³C illustrate the importance of how stress is imposed during growth.