Palatability trials on hardwood leaf litter grown under elevated CO2: a stable carbon isotope study

The palatability to isopods and microbes of a broad range of hardwood leaf litter, derived from three field CO₂-enrichment experiments in the USA, was investigated, using δ¹³C, to trace the C flow from litter to isopods and to CO₂ respired by microbial decomposition. Leaf litter grown under elevated CO₂ had δ¹³C values ranging from −39 to −45‰, which were significantly different from ambient litter δ¹³C values of around −30‰. Litter palatability to isopods of the Porcellio sp. was tested by incubating ambient- and elevated-CO₂ litter, and a mixture of the two, in the presence of isopods for 14 days, under environmentally controlled conditions; δ¹³C was measured on litter and isopods' body before and after incubation. In an additional experiment, litter was incubated in the absence of fauna for 30 days, and on five occasions the δ¹³C of the CO₂ respired from litter was measured. The ¹³C label was clearly carried from the litter source to the isopods' bodies, and their faeces. For microbial-respired CO₂, δ¹³C was significantly higher than that of the litter source, suggesting preferential degradation of substrates enriched in ¹³C as compared to those in the overall litter. With the exception of Quercus myrtifolia leaf litter, elevated CO₂ did not affect the palatability to isopods nor the microbial degradation of any of the litters, possibly as a result of unaltered litter N concentration. However, significant differences in litter palatability and decay rates were observed among the different species. With this study, the use of isotopically labelled litter material was confirmed as a key methodology that can significantly contribute to the advancement of the understanding of litter decomposition and of the quantification of C fluxes in the process.     

Carbon dioxide emissions of soils under pure and mixed stands of beech and spruce, affected by decomposing foliage litter mixtures

Soil respiration is the largest terrestrial source of CO₂ to the atmosphere. In forests, roughly half of the soil respiration is autotrophic (mainly root respiration) while the remainder is heterotrophic, originating from decomposition of soil organic matter. Decomposition is an important process for cycling of nutrients in forest ecosystems. Hence, tree species induced changes may have a great impact on atmospheric CO₂ concentrations. Since studies on the combined effects of beech-spruce mixtures are very rare, we firstly measured CO₂ emission rates in three adjacent stands of pure spruce (Picea abies), mixed spruce-beech and pure beech (Fagus sylvatica) on three base-rich sites (Flysch) and three base-poor sites (Molasse; yielding a total of 18 stands) during two summer periods using the closed chamber method. CO₂ emissions were higher on the well-aerated sandy soils on Molasse than on the clayey soils on Flysch, characterized by frequent water logging. Mean CO₂ effluxes increased from spruce (41) over the mixed (55) to the beech (59) stands on Molasse, while tree species effects were lower on Flysch (30-35, mixed > beech = spruce; all data in mg CO₂-C m⁻² h⁻¹). Secondly, we studied decomposition after fourfold litter manipulations at the 6 mixed species stands: the O_i - and O_e horizons were removed and replaced by additions of beech -, spruce - and mixed litter of the adjacent pure stands of known chemical quality and one zero addition (blank) in open rings (20 cm inner diameter), which were covered with meshes to exclude fresh litter fall. Mass loss within two years amounted to 61-68% on Flysch and 36-44% on Molasse, indicating non-additive mixed species effects (mixed litter showed highest mass loss). However, base cation release showed a linear response, increasing from the spruce - over the mixed - to the beech litter. The differences in N release (immobilization) resulted in a characteristic converging trend in C/N ratios for all litter compositions on both bedrocks during decomposition. In the summers 2006 and 2007 we measured CO₂ efflux from these manipulated areas (a closed chamber fits exactly over such a ring) as field indicator of the microbial activity. Net fluxes (subtracting the so-called blank values) are considered an indicator of litter induced changes only and increased on both bedrocks from the spruce - over the mixed - to the beech litter. According to these measurements, decomposing litter contributed between 22-32% (Flysch) and 11-28% (Molasse) to total soil respiration, strengthening its role within the global carbon cycle.     

Elevated CO2 concentration and nitrogen fertilisation effects on N2O and CH4 fluxes and biomass production of Phleum pratense on farmed peat soil

The effects of elevated CO₂ supply on N₂O and CH₄ fluxes and biomass production of Phleum pratense were studied in a greenhouse experiment. Three sets of 12 farmed peat soil mesocosms (10 cm dia, 47 cm long) sown with P. pratense and equally distributed in four thermo-controlled greenhouses were fertilised with a commercial fertiliser in order to add 2, 6 or 10 g N m⁻². In two of the greenhouses, CO₂ concentration was kept at atmospheric concentration (360 μmol mol⁻¹) and in the other two at doubled concentration (720 μmol mol⁻¹). Soil temperature was kept at 15 °C and air temperature at 20 °C. Natural lighting was supported by artificial light and deionized water was used to regulate soil moisture. Forage was harvested and the plants fertilised three times during the basic experiment, followed by an extra fertilisations and harvests. At the end of the experiment CH₄ production and CH₄ oxidation potentials were determined; roots were collected and the biomass was determined. From the three first harvests the amount of total N in the aboveground biomass was determined. N₂O and CH₄ exchange was monitored using a closed chamber technique and a gas chromatograph. The highest N₂O fluxes (on average, 255 μg N₂O m⁻² h⁻¹ during period IV) occurred just after fertilisation at high water contents, and especially at the beginning of the growing season (on average, 490 μg N₂O m⁻² h⁻¹ during period I) when the competition of vegetation for N was low. CH₄ fluxes were negligible throughout the experiment, and for all treatments the production and oxidation potentials of CH₄ were inconsequential. Especially at the highest rates of fertilisation, the elevated supply of CO₂ increased above- and below-ground biomass production, but both at the highest and lowest rates of fertilisation, decreased the total amount of N in the aboveground dry biomass. N₂O fluxes tended to be higher under doubled CO₂ concentrations, indicating that increasing atmospheric CO₂ concentration may affect N and C dynamics in farmed peat soil.     

Effects of elevated temperature, elevated CO2 and fertilization on quality and subsequent decomposition of silver birch leaf litter

We examined the quality and decomposition of naturally abscised leaves of silver birch (Betula pendula) seedlings subjected to three different levels of fertilization under ambient and elevated levels of temperature and CO₂. At the end of the second growing season, the chemical composition of the litter collected from the seedlings was analyzed. Whole-leaf samples from pooled litter from each of the four replicates from each treatment were put in mesh bags and transferred to ambient climate in the field. The remaining mass of litter was measured by sampling bags in May and October throughout the four-year incubation period. Fertilization with all nutrients decreased the initial carbon and tannin contents of litter, and increased the proportion of the fast-decomposing fraction, but still fertilization slowed down the decomposition of this fraction. Initially, the estimated proportion of the fast-decomposing fraction was smallest in elevated CO₂ + temperature, and largest in ambient climate. During decomposition, elevated growth-temperature slowed down decomposition of the fast fraction under ambient CO₂ but increased it under elevated CO₂. The changes in litter decomposition rates found over four years were not very large. However, we conclude that the interactions of different factors lead to different results than if the factors had been studied separately, and future studies should take interactions into account.     

Five years of simulated atmospheric nitrogen deposition have only subtle effects on the fate of newly synthesized carbon in Calluna vulgaris and Eriophorum vaginatum

To understand the implications of atmospheric nitrogen deposition on carbon turnover in peatlands, we conducted a ¹³C pulse labeling experiment on Calluna vulgaris and Eriophorum vaginatum already receiving long-term (5 years) amendments of 56 kg N ha⁻¹ y⁻¹ as ammonium or nitrate. We examined shoot tissue retention, net ecosystem respiration returns of the ¹³C pulse, and soil porewater DOC content under the two species. ¹³C fixation in Eriophorum leaves was enhanced with nitrogen addition and doubled with nitrate supply. This newly fixed C appeared to be relocated below-ground faster with nitrogen fertilization as respiration returns were unaffected by N inputs. By contrast, increases in ¹³C fixation were not observed in Calluna. Instead, net ecosystem respiration rates over Calluna increased with N fertilization. There was no significant label incorporation into DOC, suggesting a conservative strategy of peatland vegetation regarding allocation of C through root exudation. Greater concentrations of total DOC were identified with nitrate addition in Calluna. Given the long-term nature of the experiment and the high N inputs, the overall impacts of nitrogen amendments on the fate of recently synthesized C in Eriophorum and Calluna in this ombrotrophic peatland were surprisingly more moderate than originally hypothesized. This may be due to N being effectively retained within the bryophyte layer, thus limiting, and delaying the onset of, below-ground effects.     

Elevated CO2, but not defoliation, enhances N cycling and increases short-term soil N immobilization regardless of N addition in a semiarid grassland

Elevated CO₂ and defoliation effects on nitrogen (N) cycling in rangeland soils remain poorly understood. Here we tested whether effects of elevated CO₂ (720 μl L⁻¹) and defoliation (clipping to 2.5 cm height) on N cycling depended on soil N availability (addition of 1 vs. 11 g N m⁻²) in intact mesocosms extracted from a semiarid grassland. Mesocosms were kept inside growth chambers for one growing season, and the experiment was repeated the next year. We added ¹⁵N (1 g m⁻²) to all mesocosms at the start of the growing season. We measured total N and ¹⁵N in plant, soil inorganic, microbial and soil organic pools at different times of the growing season. We combined the plant, soil inorganic, and microbial N pools into one pool (PIM-N pool) to separate biotic + inorganic from abiotic N residing in soil organic matter (SOM). With the ¹⁵N measurements we were then able to calculate transfer rates of N from the active PIM-N pool into SOM (soil N immobilization) and vice versa (soil N mobilization) throughout the growing season. We observed significant interactive effects of elevated CO₂ with N addition and defoliation with N addition on soil N mobilization and immobilization. However, no interactive effects were observed for net transfer rates. Net N transfer from the PIM-N pool into SOM increased under elevated CO₂, but was unaffected by defoliation. Elevated CO₂ and defoliation effects on the net transfer of N into SOM may not depend on soil N availability in semiarid grasslands, but may depend on the balance of root litter production affecting soil N immobilization and root exudation affecting soil N mobilization. We observed no interactive effects of elevated CO₂ with defoliation. We conclude that elevated CO₂, but not defoliation, may limit plant productivity in the long-term through increased soil N immobilization.     

Winter soil CO2 efflux and its contribution to annual soil respiration in different ecosystems of a forest-steppe ecotone, north China

Most soil respiration measurements are conducted during the growing season. In tundra and boreal forest ecosystems, cumulative winter soil CO₂ fluxes are reported to be a significant component of their annual carbon budgets. However, little information on winter soil CO₂ efflux is known from mid-latitude ecosystems. Therefore, comparing measurements of soil respiration taken annually versus during the growing season will improve the accuracy of ecosystem carbon budgets and the response of soil CO₂ efflux to climate changes. In this study we measured winter soil CO₂ efflux and its contribution to annual soil respiration for seven ecosystems (three forests: Pinus sylvestris var. mongolica plantation, Larix principis-rupprechtii plantation and Betula platyphylla forest; two shrubs: Rosa bella and Malus baccata; and two meadow grasslands) in a forest-steppe ecotone, north China. Overall mean winter and growing season soil CO₂ effluxes were 0.15-0.26 μmol m⁻² s⁻¹ and 2.65-4.61 μmol m⁻² s⁻¹, respectively, with significant differences in the growing season among the different ecosystems. Annual Q₁₀ (increased soil respiration rate per 10 °C increase in temperature) was generally higher than the growing season Q₁₀. Soil water content accounted for 84% of the variations in growing season Q₁₀ and soil temperature range explained 88% of the variation in annual Q₁₀. Soil organic carbon density to 30 cm depth was a good surrogate for SR₁₀ (basal soil respiration at a reference temperature of 10 °C). Annual soil CO₂ efflux ranged from 394.76 g C m⁻² to 973.18 g C m⁻² using observed ecosystem-specific response equations between soil respiration and soil temperature. Estimates ranged from 424.90 g C m⁻² to 784.73 g C m⁻² by interpolating measured soil respiration between sampling dates for every day of the year and then computing the sum to obtain the annual value. The contributions of winter soil CO₂ efflux to annual soil respiration were 3.48-7.30% and 4.92-7.83% using interpolated and modeled methods, respectively. Our results indicate that in mid-latitude ecosystems, soil CO₂ efflux continues throughout the winter and winter soil respiration is an important component of annual CO₂ efflux.     

CO2 and inorganic N supply modify competition for N between co-occurring grass plants, tree seedlings and soil microorganisms

Plant-plant and plant-soil interactions play a key role in determining plant community structure and ecosystem function. However, the effects of global change on the interplay between co-occurring plants and soil microbes in successional communities are poorly understood. In this study, we investigated competition for nitrogen (N) between soil microorganisms, grass plants and establishing tree seedlings under factorial carbon dioxide (CO₂) and N treatments. Fraxinus excelsior seedlings were germinated in the presence or absence of grass competition (Dactylis glomerata) at low (380 μmol mol⁻¹) or high (645 μmol mol⁻¹) CO₂ and at two levels of N nutrition in a mesocosm experiment. Pulse ¹⁵N labelling was used to examine N partitioning among plant and soil compartments. Dactylis exerted a strong negative effect on Fraxinus biomass, N capture and ¹⁵N recovery irrespective of N and CO₂ treatment. In contrast, the presence of Dactylis had a positive effect on the microbial N pool. Plant and soil responses to N treatment were of a greater magnitude compared with responses to elevated CO₂, but the pattern of Fraxinus- and microbial-N pool response to N and CO₂ varied depending on grass competition treatment. Within the Dactylis competition treatment, decreases in Fraxinus biomass in response to N were not mirrored by decreases in tree seedling N content, suggesting a shift from below- to above-ground competition. In the Dactylis-sown pots, ¹⁵N recovery could be ranked Dactylis > microbial pool > Fraxinus in all N and CO₂ treatment combinations. Inequalities between Fraxinus and soil microorganisms in terms of ¹⁵N recovery were exacerbated by N addition. Contrary to expectations, elevated CO₂ did not increase plant-microbe competition. Nevertheless, microbial ¹⁵N recovery showed a small positive increase in the high CO₂ treatment. Overall, elevated CO₂ and N supply did not interact on plant/soil N partitioning. Our data suggest that the competitive balance between establishing tree seedlings and grass plants in an undisturbed sward is relatively insensitive to CO₂ or N-induced modifications in N competition between plant and soil compartments.     

Nitrogen fixation by biological soil crusts and heterotrophic bacteria in an intact Mojave Desert ecosystem with elevated CO2 and added soil carbon

Fixation of N by biological soil crusts and free-living heterotrophic soil microbes provides a significant proportion of ecosystem N in arid lands. To gain a better understanding of how elevated CO₂ may affect N₂-fixation in aridland ecosystems, we measured C₂H₂ reduction as a proxy for nitrogenase activity in biological soil crusts for 2 yr, and in soils either with or without dextrose-C additions for 1 yr, in an intact Mojave Desert ecosystem exposed to elevated CO₂. We also measured crust and soil δ¹⁵N and total N to assess changes in N sources, and δ¹³C of crusts to determine a functional shift in crust species, with elevated CO₂. The mean rate of C₂H₂ reduction by biological soil crusts was 76.9±5.6 μmol C₂H₄ m⁻² h⁻¹. There was no significant CO₂ effect, but crusts from plant interspaces showed high variability in nitrogenase activity with elevated CO₂. Additions of dextrose-C had a positive effect on rates of C₂H₂ reduction in soil. There was no elevated CO₂ effect on soil nitrogenase activity. Plant cover affected soil response to C addition, with the largest response in plant interspaces. The mean rate of C₂H₂ reduction in soils either with or without C additions were 8.5±3.6 μmol C₂H₄ m⁻² h⁻¹ and 4.8±2.1 μmol m⁻² h⁻¹, respectively. Crust and soil δ¹⁵N and δ¹³C values were not affected by CO₂ treatment, but did show an effect of cover type. Crust and soil samples in plant interspaces had the lowest values for both measurements. Analysis of soil and crust [N] and δ¹⁵N data with the Rayleigh distillation model suggests that any plant community changes with elevated CO₂ and concomitant changes in litter composition likely will overwhelm any physiological changes in N₂-fixation.     

Wetland macrophyte decomposition under different nutrient conditions: Relationships between decomposition rate, enzyme activities and microbial biomass

We used oligotrophic, P-limited herbaceous wetlands of northern Belize as a model system, on which to document and explain how changes in nutrient content along a salinity gradient affect activities of extracellular enzymes involved in macrophyte decomposition. To determine what is more important for decomposition, the initial litter quality, or site differences, we used reciprocal litter placement in a combined “site quality” and “litter quality” experiment running from August 2003 to April 2004. The experiment was set up in long-term control and nutrient addition plots (P, N, and NP) established in 2001 in 15 limestone-based inland marshes with a wide range of water conductivities (200-6000 μS) and a uniform pH (7.0-7.7) dominated by emergent macrophytes, Eleocharis spp. There were no differences among the plots in total sediment N and water NH₄-N, but total and KCl-extractable sediment P and water PO₄-P were significantly higher in P and NP plots throughout the duration of the experiment. The initial litter N content was slightly but significantly different between control and N plots versus P and NP plots (5.7 and 7.1 mg g⁻¹, respectively). The difference was much bigger for litter P content, 0.1 and 0.7 mg g⁻¹, respectively. Enzyme activities of alkaline phosphatase, leucine-aminopeptidase, arylsulfatase, and β-glucosidase were measured fluorometrically in Eleocharis litter in both the litterbag experiment and the naturally decomposing material. Total phospholipid fatty acid (PLFA) content in litter samples was used as a measure of microbial biomass present. Phosphatase always exhibited the highest activity of the enzymes studied, followed by leucine-aminopeptidase, arylsulfatase and β-glucosidase. There were no significant differences between enzyme activities from litterbags and the unconfined litter. Phosphatase activity was significantly suppressed in P-addition plots under all salinity levels while the activities of the remaining enzymes were significantly higher in P-enriched plots. There was a strong correlation between decomposition coefficient k-values and most of the enzymes as well as between the amount of PLFA and enzyme activities. PLFA, arylsulfatase, and litter C/P were the best predictors of k-values.