Linking microbial activity and soil organic matter transformations in forest soils under elevated CO2

Soil organic matter (SOM) dynamics ultimately govern the ability of soil to provide long‐term C sequestration and the nutrients required for ecosystem productivity. Predicting belowground responses to elevated CO₂ requires an integrated understanding of SOM transformations and the microbial activity that governs them. It remains unclear how the microorganisms upon which these transformations depend will function in an elevated CO₂ world. This study examines SOM transformations and microbial metabolism in soils from the Duke Free Air Carbon Enrichment site in North Carolina, USA. We assessed microbial respiration and net nitrogen (N) mineralization in soils with and without elevated CO₂ exposure during a 100‐day incubation. We also traced the depleted C isotopic signature of the supplemental CO₂ into SOM and the soils' phospholipid fatty acids (PLFA), which serve as biomarkers for living cells. Cumulative net N mineralization in elevated CO₂ soils was 50% that in control soils after a 100‐day incubation. Respiration was not altered with elevated CO₂. C : N ratios of bulk SOM did not change with elevated CO₂, but incubation data suggest that the C : N ratios of mineralized organic matter increased with elevated CO₂. Values of SOM δ¹³C were depleted with elevated CO₂ (?26.7±0.2 vs. ?30.2±0.3‰), reflecting the depleted signature of the supplemental CO₂. We compared δ¹³C of individual PLFA with the δ¹³C of SOM to discern incorporation of the depleted C isotopic signature into soil microbial groups in elevated CO₂ plots. PLFA i15:0, a15:0, and 10Met18:0 reflected significant incorporation of recently produced photosynthate, suggesting that the bacterial groups defined by these biomarkers are active metabolizers in elevated CO₂ soils. At least one of these groups (actinomycetes, 10Met18:0) specializes in metabolizing less labile substrates. Because control plots did not receive an equivalent ¹³C tracer, we cannot determine from these data whether this group of organisms was stimulated by elevated CO₂ compared with these organisms in control soils. Stimulation of this group, if it occurred in the elevated CO₂ plot, would be consistent with a decline in the availability of mineralizable organic matter with elevated CO₂, which incubation data suggest may be the case in these soils.     

Diffusive fractionation complicates isotopic partitioning of autotrophic and heterotrophic sources of soil respiration

Carbon isotope ratios (δ¹³C) of heterotrophic and rhizospheric sources of soil respiration under deciduous trees were evaluated over two growing seasons. Fluxes and δ¹³C of soil respiratory CO₂ on trenched and untrenched plots were calculated from closed chambers, profiles of soil CO₂ mole fraction and δ¹³C and continuous open chambers. δ¹³C of respired CO₂ and bulk carbon were measured from excised leaves and roots and sieved soil cores. Large diel variations (>5‰) in δ¹³C of soil respiration were observed when diel flux variability was large relative to average daily fluxes, independent of trenching. Soil gas transport modelling supported the conclusion that diel surface flux δ¹³C variation was driven by non‐steady state gas transport effects. Active roots were associated with high summertime soil respiration rates and around 1‰ enrichment in the daily average δ¹³C of the soil surface CO₂ flux. Seasonal δ¹³C variability of about 4‰ (most enriched in summer) was observed on all plots and attributed to the heterotrophic CO₂ source.     

Decomposition of soil and plant carbon from pasture systems after 9 years of exposure to elevated CO2: impact on C cycling and modeling

Elevated atmospheric CO₂ may alter decomposition rates through changes in plant material quality and through its impact on soil microbial activity. This study examines whether plant material produced under elevated CO₂ decomposes differently from plant material produced under ambient CO₂. Moreover, a long‐term experiment offered a unique opportunity to evaluate assumptions about C cycling under elevated CO₂ made in coupled climate–soil organic matter (SOM) models. Trifolium repens and Lolium perenne plant materials, produced under elevated (60 Pa) and ambient CO₂ at two levels of N fertilizer (140 vs. 560 kg ha^?1 yr^?1), were incubated in soil for 90 days. Soils and plant materials used for the incubation had been exposed to ambient and elevated CO₂ under free air carbon dioxide enrichment conditions and had received the N fertilizer for 9 years. The rate of decomposition of L. perenne and T. repens plant materials was unaffected by elevated atmospheric CO₂ and rate of N fertilization. Increases in L. perenne plant material C : N ratio under elevated CO₂ did not affect decomposition rates of the plant material. If under prolonged elevated CO₂ changes in soil microbial dynamics had occurred, they were not reflected in the rate of decomposition of the plant material. Only soil respiration under L. perenne, with or without incorporation of plant material, from the low‐N fertilization treatment was enhanced after exposure to elevated CO₂. This increase in soil respiration was not reflected in an increase in the microbial biomass of the L. perenne soil. The contribution of old and newly sequestered C to soil respiration, as revealed by the ¹³C‐CO₂ signature, reflected the turnover times of SOM–C pools as described by multipool SOM models. The results do not confirm the assumption of a negative feedback induced in the C cycle following an increase in CO₂, as used in coupled climate–SOM models. Moreover, this study showed no evidence for a positive feedback in the C cycle following additional N fertilization.     

Soil carbon pool structure and temperature sensitivity inferred using CO2 and 13CO2 incubation fluxes from five Hawaiian soils

We measured respiration and ¹³C values of respiredand soil carbon in long-term incubations of soils from two forests andthree pastures along an altitudinal gradient in Hawaii. CO₂fluxes early in the incubations decreased rapidly, and then stabilizedat approximately 20% of initial values for sevenmonths. We suggest that the rapid drop and subsequent stabilizationof respiration reflects a change in the dominant source of theCO₂ from labile (active) to much more recalcitrantpools of soil organic matter (SOM). Estimates of active SOM weremade by integrating all of the carbon respired in excess of thatattributable to respiration of the intermediate SOM pool; thesevalues ranged from 0.7–4.3% of total soil C.¹³C values for carbon respired from the pasturesoils showed that older, forest-derived C contributed an increasingfraction of total soil respiration with time. Initial and late-stagerespiration responded similarly to changes in temperature, suggestingthat intermediate SOM is as sensitive to temperature as the activefraction.     

14C – a tool for separation of autotrophic and heterotrophic soil respiration

We assessed the potential of using ¹⁴C contents of soil respired CO₂ to calculate the contributions of heterotrophic and autotrophic respiration to total soil respiration. The partitioning of these fluxes is of utmost importance to evaluate implications of environmental change on soil carbon cycling and sequestration. At three girdled forest stands in Sweden and Germany, where the tree root (autotrophic) respiration had been eliminated, we measured both flux rates and ¹⁴C contents of soil respired CO₂ in girdled and control plots in the summers of 2001 or 2002. At all stands, CO₂ flux rates were slightly higher in the control plots, whereas the ¹⁴C contents of respired CO₂ tended to be higher in the girdled plots. This was expected and confirmed that heterotrophically respired CO₂ cycles more slowly through the forest ecosystem than autotrophically respired CO₂. On the basis of these data, the contributions of hetero‐ and autotrophic respiration to total soil respiration were calculated using two separate approaches (i.e. based on flux rates or ¹⁴C). Fractions of heterotrophic respiration ranged from 53% to 87%. Values calculated by both approaches did not differ significantly from each other. Finally, we compared the ¹⁴C contents of soil respired CO₂ in the girdled plots with the ¹⁴C contents of heterotrophically respired CO₂ calculated by three different ¹⁴C models. None of the models matched the measured data sufficiently. In addition, we suspect that inherent effects of girdling may cause the ¹⁴C content of CO₂ respired in the girdled plots to be lower than ‘true’ heterotrophically respired CO₂ in an undisturbed plot. Nevertheless, we argue that measurements and modeling of ¹⁴C can be developed into a valuable tool for separating heterotrophic and autotrophic soil respiration (e.g. when girdling cannot be performed).     

Immobilization, stabilization and remobilization of nitrogen in forest soils at elevated CO2: a 15N and 13C tracer study

The fate of immobilized N in soils is one of the great uncertainties in predicting C sequestration at increased CO₂ and N deposition. In a dual isotope tracer experiment (¹³C, ¹⁵N) within a 4‐year CO₂ enrichment (+200 ppm_v) study with forest model ecosystems, we (i) quantified the effects of elevated CO₂ on the partitioning of N; (ii) traced immobilized N into physically separated pools of soil organic matter (SOM) with turnover rates known from their ¹³C signals; and (iii) estimated the remobilization and thus, the bio‐availability of newly sequestered C and N. (1) CO₂ enrichment significantly decreased NO₃^? concentrations in soil waters and export from 1.5 m deep lysimeters by 30–80%. Consequently, elevated CO₂ increased the overall retention of N in the model ecosystems. (2) About 60–80% of added ¹⁵NH₄¹⁵NO₃ were retained in soils. The clay fraction was the greatest sink for the immobilized ¹⁵N sequestering 50–60% of the total new soil N. SOM associated with clay contained only 25% of the total new soil C pool and had small C/N ratios (<13), indicating that it consists of humified organic matter with a relatively slow turn over rate. This implies that added ¹⁵N was mainly immobilized in stable mineral‐bound SOM pools. (3) Incubation of soils for 1 year showed that the remobilization of newly sequestered N was three to nine times smaller than that of newly sequestered C. Thus, inorganic inputs of N were stabilized more effectively in soils than C. Significantly less newly sequestered N was remobilized from soils previously exposed to elevated CO₂. In summary, our results show firstly that a large fraction of inorganic N inputs becomes effectively immobilized in relative stable SOM pools and secondly that elevated CO₂ can increase N retention in soils and hence it may tighten N cycling and diminish the risk of nitrate leaching to groundwater.     

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].     

Environmental and physiological controls on the carbon isotope composition of CO2 respired by leaves and roots of a C3 woody legume (Prosopis velutina) and a C4 perennial grass (Sporobolus wrightii)

Accurate estimates of the δ¹³C value of CO₂ respired from roots (δ¹³C_{R_root}) and leaves (δ¹³C_{R_leaf}) are important for tracing and understanding changes in C fluxes at the ecosystem scale. Yet the mechanisms underlying temporal variation in these isotopic signals are not fully resolved. We measured δ¹³C_{R_leaf}, δ¹³C_{R_root}, and the δ¹³C values and concentrations of glucose and sucrose in leaves and roots in the C₄ grass Sporobolus wrightii and the C₃ tree Prosopis velutina in a savanna ecosystem in southeastern Arizona, USA. Night‐time variation in δ¹³C_{R_leaf} of up to 4.6 ± 0.6‰ in S. wrightii and 3.0 ± 0.6‰ in P. velutina were correlated with shifts in leaf sucrose concentration, but not with changes in δ¹³C values of these respiratory substrates. Strong positive correlations between δ¹³C_{R_root} and root glucose δ¹³C values in P. velutina suggest large diel changes in δ¹³C_{R_root} (were up to 3.9‰) influenced by short‐term changes in δ¹³C of leaf‐derived phloem C. No diel variation in δ¹³C_{R_root} was observed in S. wrightii. Our findings show that short‐term changes in δ¹³C_{R_leaf} and δ¹³C_{R_root} were both related to substrate isotope composition and concentration. Changes in substrate limitation or demand for biosynthesis may largely control short‐term variation in the δ¹³C of respired CO₂ in these species.     

δ13C of CO2 respired in the dark in relation to δ13C of leaf metabolites: comparison between Nicotiana sylvestris and Helianthus annuus under drought

The variations of δ¹³C in leaf metabolites (lipids, organic acids, starch and soluble sugars), leaf organic matter and CO₂ respired in the dark from leaves of Nicotiana sylvestris and Helianthus annuus were investigated during a progressive drought. Under well‐watered conditions, CO₂ respired in the dark was ¹³C‐enriched compared to sucrose by about 4‰ in N. sylvestris and by about 3‰ and 6‰ in two different sets of experiments in H. annuus plants. In a previous work on cotyledonary leaves of Phaseolus vulgaris, we observed a constant ¹³C‐enrichment by about 6‰ in respired CO₂ compared to sucrose, suggesting a constant fractionation during dark respiration, whatever the leaf age and relative water content. In contrast, the ¹³C‐enrichment in respired CO₂ increased in dehydrated N. sylvestris and decreased in dehydrated H. annuus in comparison with control plants. We conclude that (i) carbon isotope fractionation during dark respiration is a widespread phenomenon occurring in C₃ plants, but that (ii) this fractionation is not constant and varies among species and (iii) it also varies with environmental conditions (water deficit in the present work) but differently among species. We also conclude that (iv) a discrimination during dark respiration processes occurred, releasing CO₂ enriched in ¹³C compared to several major leaf reserves (carbohydrates, lipids and organic acids) and whole leaf organic matter.     

Chemistry and decomposition of litter from Populus tremuloides Michaux grown at elevated atmospheric CO2 and varying N availability

It has been hypothesized that greater production of total nonstructural carbohydrates (TNC) in foliage grown under elevated atmospheric carbon dioxide (CO₂) will result in higher concentrations of defensive compounds in tree leaf litter, possibly leading to reduced rates of decomposition and nutrient cycling in forest ecosystems of the future. To evaluate the effects of elevated atmospheric CO₂ on litter chemistry and decomposition, we performed a 111 day laboratory incubation with leaf litter of trembling aspen (Populus tremuloides Michaux) produced at 36 Pa and 56 Pa CO₂ and two levels of soil nitrogen (N) availability. Decomposition was quantified as microbially respired CO₂ and dissolved organic carbon (DOC) in soil solution, and concentrations of nonstructural carbohydrates, N, carbon (C), and condensed tannins were monitored throughout the incubation. Growth under elevated atmospheric CO₂ did not significantly affect initial litter concentrations of TNC, N, or condensed tannins. Rates of decomposition, measured as both microbially respired CO₂ and DOC did not differ between litter produced under ambient and elevated CO₂. Total C lost from the samples was 38 mg g^?1 litter as respired CO₂ and 138 mg g^?1 litter as DOC, suggesting short‐term pulses of dissolved C in soil solution are important components of the terrestrial C cycle. We conclude that litter chemistry and decomposition in trembling aspen are minimally affected by growth under higher concentrations of CO₂.     

Estimating the contribution of leaf litter decomposition to soil CO2 efflux in a beech forest using 13C-depleted litter

The contribution of leaf litter decomposition to total soil CO₂ efflux (F_L/F) was evaluated in a beech (Fagus sylvatica L.) forest in eastern France. The Keeling‐plot approach was applied to estimate the isotopic composition of respired soil CO₂ from soil covered with either control (?30.32‰) or ¹³C‐depleted leaf litter (?49.96‰). The δ¹³C of respired soil CO₂ ranged from ?25.50‰ to ?22.60‰ and from ?24.95‰ to ?20.77‰, respectively, with depleted or control litter above the soil. The F_L/F ratio was calculated by a single isotope linear mixing model based on mass conservation equations. It showed seasonal variations, increasing from 2.8% in early spring to about 11.4% in mid summer, and decreasing to 4.2% just after leaf fall. Between December 2001 and December 2002, cumulated F and F_L reached 0.98 and 0.08 kg_C m^?2, respectively. On an annual basis, decomposition of fresh leaf litter accounted for 8% of soil respiration and 80% of total C loss from fresh leaf litter. The other fraction of carbon loss during leaf litter decomposition that is assumed to have entered the soil organic matter pool (i.e. 20%) represents only 0.02 kg_C m^?2.     

Hair of grazing cattle provides an integrated measure of the effects of site conditions and interannual weather variability on δ13C of temperate humid grassland

The carbon isotope composition (δ¹³C) of C3 ecosystems is sensitive to water availability, and provides important information for the assessment of terrestrial carbon (C) sink/source activity. Here, we report the effects of plant available soil water (PAW) on community ¹³C signatures of temperate humid grassland. The 5‐year study was conducted on pastures exhibiting a large range of PAW capacity that were located on two site types: peat and mineral soils. The data set included the centennial drought year 2003, and data from wet years (2000 and 2002). Seasonal variation of PAW was modeled using PAW capacity of each pasture, precipitation inputs and evapotranspiration estimates. Community ¹³C signatures were derived from the δ¹³C of vegetation and segments of tail switch hair of cattle grown while grazing pastures. Hair ¹³C signatures provided an assimilation‐weighted ¹³C signal that integrated both spatial (paddock‐scale) and temporal (grazing season) variation of ¹³C signatures on a pasture. The δ¹³C of hair and vegetation increased with decreasing modeled PAW in the same way on mineral and peat soils. But, at a given PAW, the δ¹³C of hair was 2.6‰ less negative than that of vegetation, reflecting the diet‐hair isotopic shift. Furthermore, the δ¹³C of hair and vegetation on peat soil pastures was 0.5‰ more negative than on pastures situated on mineral soil. This may have resulted from a ～10 ppm CO₂ enrichment of canopy air derived from ongoing peat mineralization. Community‐scale season‐mean ¹³C discrimination (Δ) exhibited a saturation‐type response towards season‐mean modeled PAW (r²=0.78), and ranged between 19.8‰ on soils with low PAW capacity during the drought year of 2003, and 21.4‰ on soils with high PAW capacity in a wet year. This indicated relatively small variation in season‐mean assimilation‐weighted p_i/p_a (0.68–0.75) between contrasting sites and years. However, this range is similar to that reported in other studies, which encompass the range from subtropical arid to humid temperate grassland. Furthermore, the tight relationship between season‐mean Δ and modeled mean PAW suggests that PAW may be used as proxy for Δ.     

Changes in the abundance of C3/C4 species of Inner Mongolia grassland: evidence from isotopic composition of soil and vegetation

Global warming, increasing CO₂ concentration, and environmental disturbances affect grassland communities throughout the world. Here, we report on variations in the C3/C4 pattern of Inner Mongolian grassland derived from soil and vegetation. Soil samples from 149 sites covering an area of approximately 250 000 km² within Inner Mongolia, People's Republic of China were analyzed for the isotopic composition (δ¹³C) of soil organic carbon (SOC). The contrast in δ¹³C between C3 and C4 plants allowed for calculation of the C3/C4 ratio from δ¹³C of SOC with a two‐member mixing model, which accounted for influences of aridity and altitude on δ¹³C of the C3 end‐member and for changes in δ¹³C of atmospheric CO₂. Maps were created geostatistically, and showed a substantially lower C4 abundance in soil than in recent vegetation (?10%). The difference between soil and vegetation varied regionally and was most pronounced within an E–W belt along 44°N and in a mountainous area, suggesting a spread of C4 plants toward northern latitudes (about 1°) and higher altitudes. The areas of high C4 abundance for present vegetation and SOC were well delineated by the isotherms of crossover temperature based on the climatic conditions of the respective time periods. Our study indicates that change in the patterns of C3/C4 composition in the Inner Mongolia grassland was mainly triggered by increasing temperature, which overrode the antagonistic effect of rising CO₂ concentrations.     

Short-term variation in the isotopic composition of organic matter allocated from the leaves to the stem of Pinus sylvestris: effects of photosynthetic and postphotosynthetic carbon isotope fractionation

We aimed to quantify the separate effects of photosynthetic and postphotosynthetic carbon isotope discrimination on δ¹³C of the fast‐turn‐over carbon pool (water soluble organic carbon and CO₂ emitted from heterotrophic tissues), including their diel variation, along the pathway of carbon transport from the foliage to the base of the stem. For that purpose, we determined δ¹³C in total and water‐soluble organic matter of the foliage plus δ¹³C and δ¹⁸O in phloem organic matter of twigs and at three heights along the stem of Pinus sylvestris over a nine‐day period, including four measurements per day. These data were related to meteorological and photosynthesis parameters and to the δ¹³C of stem‐emitted CO₂. In the canopy (foliage and twigs), the δ¹³C of soluble organic matter varied diurnally with amplitudes of up to 1.9‰. The greatest ¹³C enrichment was recorded during the night/early morning, indicating a strong influence of starch storage and remobilization on the carbon isotope signatures of sugars exported from the leaves. ¹³C enrichment of soluble organic matter from the leaves to the twig phloem and further on to the phloem of the stem was supposed to be a result of carbon isotope fractionation associated with metabolic processes in the source and sink tissues. CO₂ emitted from the stem was enriched by 2.3–5.2‰ compared with phloem organic matter. When day‐to‐day variation was addressed, water‐soluble leaf δ¹³C and twig phloem δ¹⁸O were strongly influenced by c_i/c_a and stomatal conductance (G_s), respectively. These results show that both photosynthetic and postphotosynthetic carbon isotope fractionation influence δ¹³C of organic matter over time, and over the length of the basipetal transport pathway. Clearly, these influences on the δ¹³C of respired CO₂ must be considered when using the latter for partitioning of ecosystem CO₂ fluxes or when the assessment of δ¹³C in organic matter is applied to estimate environmental effects in c_i/c_a.     

Canopy-scale δ13C of photosynthetic and respiratory CO2 fluxes: observations in forest biomes across the United States

The δ¹³C values of atmospheric carbon dioxide (CO₂) can be used to partition global patterns of CO₂ source/sink relationships among terrestrial and oceanic ecosystems using the inversion technique. This approach is very sensitive to estimates of photosynthetic ¹³C discrimination by terrestrial vegetation (Δ_A), and depends on δ¹³C values of respired CO₂ fluxes (δ¹³C_R). Here we show that by combining two independent data streams – the stable isotope ratios of atmospheric CO₂ and eddy‐covariance CO₂ flux measurements – canopy scale estimates of Δ_A can be successfully derived in terrestrial ecosystems. We also present the first weekly dataset of seasonal variations in δ¹³C_R from dominant forest ecosystems in the United States between 2001 and 2003. Our observations indicate considerable summer‐time variation in the weekly value of δ¹³C_R within coniferous forests (4.0‰ and 5.4‰ at Wind River Canopy Crane Research Facility and Howland Forest, respectively, between May and September). The monthly mean values of δ¹³C_R showed a smaller range (2–3‰), which appeared to significantly correlate with soil water availability. Values of δ¹³C_R were less variable during the growing season at the deciduous forest (Harvard Forest). We suggest that the negative correlation between δ¹³C_R and soil moisture content observed in the two coniferous forests should represent a general ecosystem response to the changes in the distribution of water resources because of climate change. Shifts in δ¹³C_R and Δ_A could be of sufficient magnitude globally to impact partitioning calculations of CO₂ sinks between oceanic and terrestrial compartments.     

An oxygen-mediated positive feedback between elevated carbon dioxide and soil organic matter decomposition in a simulated anaerobic wetland

We examined the effects of elevated atmospheric CO₂ on soil carbon decomposition in an experimental anaerobic wetland system. Pots containing either bare C₄‐derived soil or the C₃ sedge Scirpus olneyi planted in C₄‐derived soil were incubated in greenhouse chambers at either ambient or twice‐ambient atmospheric CO₂. We measured CO₂ flux from each pot, quantified soil organic matter (SOM) mineralization using δ¹³C, and determined root and shoot biomass. SOM mineralization increased in response to elevated CO₂ by 83–218% (P<0.0001). In addition, soil redox potential was significantly and positively correlated with root biomass (P= 0.003). Our results (1) show that there is a positive feedback between elevated atmospheric CO₂ concentrations and wetland SOM decomposition and (2) suggest that this process is mediated by the release of oxygen from the roots of wetland plants. Because this feedback may occur in any wetland system, including peatlands, these results suggest a limitation on the size of the carbon sink presented by anaerobic wetland soils in a future elevated‐CO₂ atmosphere.     

Keeling plots for hummingbirds: a method to estimate carbon isotope ratios of respired CO<Subscript>2</Subscript> in small vertebrates

The carbon isotope composition of an animals breath reveals the composition of the nutrients that it catabolizes for energy. Here we describe the use of Keeling plots, a method widely applied in ecosystem ecology, to measure the ¹³C of respired CO₂ of small vertebrates. We measured the ¹³C of Rufous Hummingbirds (Selasphorus rufus) in the laboratory and of Mourning (Zenaida macroura) and White-winged (Z. asiatica) Doves in the field. In the laboratory, when hummingbirds were fed a sucrose based C3 diet, the ¹³C of respired CO₂ was not significantly different from that of their diet (¹³C_{C3 diet}). The ¹³C of respired CO₂ for C3 fasted birds was slightly, albeit significantly, depleted in ¹³C relative to ¹³C_{C3 diet}. Six hours after birds were shifted to a sucrose based C4 diet, the isotopic composition of their breath revealed that birds were catabolizing a mixture of nutrients derived from both the C3 and the C4 diet. In the field, the ¹³C of respired CO₂ from Mourning and White-winged Doves reflected that of their diets: the CAM saguaro cactus (Carnegeia gigantea) and C3 seeds, respectively. Keeling plots are an easy, effective and inexpensive method to measure ¹³C of respired CO₂ in the lab and the field.     

On the 13C/12C isotopic signal of day and night respiration at the mesocosm level

While there is currently intense effort to examine the ¹³C signal of CO₂ evolved in the dark, less is known on the isotope composition of day‐respired CO₂. This lack of knowledge stems from technical difficulties to measure the pure respiratory isotopic signal: day respiration is mixed up with photorespiration, and there is no obvious way to separate photosynthetic fractionation (pure c_i/c_a effect) from respiratory effect (production of CO₂ with a different δ¹³C value from that of net‐fixed CO₂) at the ecosystem level. Here, we took advantage of new simple equations, and applied them to sunflower canopies grown under low and high [CO₂]. We show that whole mesocosm‐respired CO₂ is slightly ¹³C depleted in the light at the mesocosm level (by 0.2–0.8‰), while it is slightly ¹³C enriched in darkness (by 1.5–3.2‰). The turnover of the respiratory carbon pool after labelling appears similar in the light and in the dark, and accordingly, a hierarchical clustering analysis shows a close correlation between the ¹³C abundance in day‐ and night‐evolved CO₂. We conclude that the carbon source for respiration is similar in the dark and in the light, but the metabolic pathways associated with CO₂ production may change, thereby explaining the different ¹²C/¹³C respiratory fractionations in the light and in the dark.     

REVIEW: Time lag between photosynthesis and carbon dioxide efflux from soil: a review of mechanisms and controls

CO₂ efflux from soil depends on the availability of organic substances respired by roots and microorganisms. Therefore, photosynthetic activity supplying carbohydrates from leaves to roots and rhizosphere is a key driver of soil CO₂. This fact has been overlooked in most soil CO₂ studies because temperature variations are highly correlated with solar radiation and mask the direct effect of photosynthesis on substrate availability in soil. This review highlights the importance of photosynthesis for rhizosphere processes and evaluates the time lag between carbon (C) assimilation and CO₂ release from soil. Mechanisms and processes contributing to the lag were evaluated. We compared the advantages and shortcomings of four main approaches used to estimate this time lag: (1) interruption of assimilate flow from leaves into the roots and rhizosphere, and analysis of the decrease of CO₂ efflux from soil, (2) time series analysis (TSA) of CO₂ fluxes from soil and photosynthesis proxies, (3) analysis of natural δ¹³C variation in CO₂ with photosynthesis‐related parameters or δ¹³C in the phloem and leaves, and (4) pulse labeling of plants in artificial ¹⁴CO₂ or ¹³CO₂ atmosphere with subsequent tracing of ¹⁴C or ¹³C in CO₂ efflux from soil. We concluded that pulse labeling is the most advantageous approach. It allows clear evaluation not only of the time lag, but also of the label dynamics in soil CO₂, and helps estimate the mean residence time of recently assimilated C in various above‐ and belowground C pools. The impossibility of tracing the phloem pressure–concentration waves by labeling approach may be overcome by its combination with approaches based on TSA of CO₂ fluxes and its δ¹³C with photosynthesis proxies. Numerous studies showed that the time lag for grasses is about 12.5±7.5 (SD) h. The time lag for mature trees was much longer (～4–5 days). Tree height slightly affected the lag, with increasing delay of 0.1 day m^?1. By evaluating bottle‐neck processes responsible for the time lag, we conclude that, for trees, the transport of assimilates in phloem is the rate‐limiting step. However, it was not possible to predict the lag based on the phloem transport rates reported in the literature. We conclude that studies of CO₂ fluxes from soil, especially in ecosystems with a high contribution of root‐derived CO₂, should consider photosynthesis as one of the main drivers of C fluxes. This calls for incorporating photosynthesis in soil C turnover models.     

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.