Does soil aging affect the N2O mitigation potential of biochar? A combined microcosm and field study

The application of biochar as a soil amendment to improve soil fertility has been suggested as a tool to reduce soil‐borne CO₂ and non‐CO₂ greenhouse gas emissions, especially nitrous oxide (N₂O). Both laboratory and field trials have demonstrated N₂O emission reduction by biochar amendment, but the long‐term effect (>1 year) has been questioned. Here, we present results of a combined microcosm and field study using a powdered beech wood biochar from slow pyrolysis. The field experiment showed that both CO₂ and N₂O emissions were still effectively reduced by biochar in the third year after application. However, biochar did not influence the biomass yield of sunflower for biogas production (Helianthus annuus L.). Biochar reduced bulk density and increased soil aeration and thus reduced the water‐filled pore space (WFPS) in the field, but was also able to suppress N₂O emission in the microcosms experiment conducted at constant WFPS. For both experiments, biochar had limited impact on soil mineral nitrogen speciation, but it reduced the accumulation of nitrite in the microcosms. Extraction of soil DNA and quantification of functional marker genes by quantitative polymerase chain reaction showed that biochar did not alter the abundance of nitrogen‐transforming bacteria and archaea in both field and microcosm experiments. In contradiction to previous experiments, this study demonstrates the long‐term N₂O emission suppression potential of a wood biochar and thus highlights its overall climate change mitigation potential. While a detailed understanding of the underlying mechanisms requires further research, we provide evidence for a range of biochar‐induced changes to the soil environment and their change with time that might explain the often observed N₂O emission suppression.     

Survival, metabolic activity and conjugative interactions of indigenous and introduced streptomycete strains in soil microcosms

The growth and activity of introduced (S. lividans TK24 pIJ673 and S.lividans TK23) and indigenous (S.griseus CAG17) streptomycete strains in soil was studied, under controlled conditions. The effect of environmental parameters such as temperature, soil water content and nutrient availability on the growth and activity of these strains, was studied using a highly dynamic fed-batch soil microcosm system. Using this new system, repeated cycles of active streptomycete growth were achieved, allowing long-term investigation of metabolic activity, plasmid stability and conjugative plasmid transfer. In long-term experiments, respiration rates and enzyme activity patterns matched the pattern of germination/sporulation cycles of the inoculants. In situ hybridisation, using fluorescently labelled oligonucleotides, also proved the presence of metabolically active streptomycete mycelia in sterile soil. Plasmid stability under varying temperatures and selective pressure was studied using the above system. In both sterile and non sterile amended antibiotic containing soil, no intraspecific transfer of plasmid pIJ673 from S.lividans TK24 to S.griseus CAG17 was detected. The soil microcosm system used, though, permitted detection of intraspecific conjugative transfer of this plasmid from S.lividans TK24 to S.lividans TK23 in soil.     

Controlled environment soil‐core microcosm unit for investigating fate,migration, and transformation of chemicals in soils

The controlled environment soil‐core microcosm unit (CESMU) methods embody a collection of techniques that began with soil sampling in the field and continued throughout the laboratory investigation of chemical fate, migration, and transformation in site‐specific soils; it was a cost‐effective investigative methodology that could be used to screen chemical materials before initiating high‐cost environmental field studies. Intact soil cores were collected in the field using a hydraulically controlled probe, delivering intact soil‐cores with minimal disturbance directly into high‐density polyethylene pipe (10.3‐cm ID). The inert polyethylene pipe was an effective hydrophobic barrier that remained an integral part of the soil‐core column, obviating subsequent transfers of soil. In the laboratory, each soil column was fitted with a porous ceramic plate and a polyethylene endcap containing fittings for teflon tubing, so that a tension could be applied at the bottom of each soil column (30–35 kPa) to mimic field conditions, thus preventing the undue buildup of water within columns that otherwise would change the chemical, physical, and biological properties of the soil. The intact soil‐cores were housed in the CESMU chamber, a controlled temperature unit with sufficient capacity for maintaining constant temperature within entire soil‐cores. Synthetic rain was added twice a week by peristaltic pump at rates simulating rainfall. Leachates were collected under tension via teflon tubing into flasks in darkness and kept at soil column temperature inside CESMU until harvested for analyses. Soil columns were harvested at intervals for sectioning by depth, extraction, and soil analyses. CESMU methods are applicable to investigations of water movement, soil chemistry, solute transport/transformation, and effects on plants.     

Microcosm responses to environmental perturbants

A flow-through microcosm facility using unfiltered ocean source water is described. “Stagnation” cleaning has proven to be a simple and effective way of maintaining seawater distribution systems free of fouling organisms and is recommended. A valveless system for regulating water flow is also recommended. The microcosm facility has been used experimentally as an extension of field surveys being conducted in Hawaiian harbors. Examples of two essentially different types of microcosm experiments are presented to illustrate this interactive approach to environmental research. One approach disturbs an established microcosm community with specific environmental perturbants and follows both the community response and recovery over prolonged time periods. Examples of perturbation by elevated nutrients and by elevated copper concentrations are given. The ability of reef flat communities to trap and retain phosphorus even in a flow-through system is demonstrated. Added copper is shown to result in a number of responses not seen in the legislatively-preferred copper toxicity tests. The second approach involves transferring substrates from the field into the microcosm and following response with time. Again, two examples are given—one involving the transfer of fouling panels, the other, of sediments. Profound changes are seen when fouling panels are transferred from a high-nutrient into a low-nutrient environment. Nutrient-rich sediments, however, are shown not to influence water-column nutrient concentrations in the microcosms, but do acquire increased infaunal populations over a 3-month period. Finally, an example of how the findings of the microcosm experiments are applied in the field is provided. The case is made that although this approach necessarily involves multivariate experimental preparations which sometimes lack precision, it is nevertheless a required and fruitful procedure in the search for better understanding of the environmental dynamics of harbor communities.     

Influence of temperature and soil drying on respiration of individual roots in citrus: integrating greenhouse observations into a predictive model for the field

In citrus, the majority of fine roots are distributed near the soil surface – a region where conditions are frequently dry and temperatures fluctuate considerably. To develop a better understanding of the relationship between changes in soil conditions and a plant’s below‐ground respiratory costs, the effects of temperature and soil drying on citrus root respiration were quantified in controlled greenhouse experiments. Chambers designed for measuring the respiration of individual roots were used. Under moist soil conditions, root respiration in citrus increased exponentially with changes in soil temperature (Q₁₀ = 1·8–2·0), provided that the changes in temperature were short‐term. However, when temperatures were held constant, root respiration did not increase exponentially with increasing temperatures. Instead, the roots acclimated to controlled temperatures above 23 °C, thereby reducing their metabolism in warmer soils. Under drying soil conditions, root respiration decreased gradually beginning at 6% soil water content and reached a minimum at <2% soil water content in sandy soil. A model was constructed from greenhouse data to predict diurnal patterns of fine root respiration based on temperature and soil water content. The model was then validated in the field using data obtained by CO₂ trapping on root systems of mature citrus trees. The trees were grown at a site where the soil temperature and water content were manipulated. Respiration predicted by the model was in general agreement with observed rates, which indicates the model may be used to estimate entire root system respiration for citrus.     

Pelagic Diatom Populations in Lentic Freshwater Microcosms

Three experiments, involving simultaneous monitoring of selected biological and chemical parameters in 50 l laboratory microcosms and the epilimnia of their parent reservoirs, were carried out from the autumn of 1978 to the winter of 1980. Experiments lasted 8-13 weeks, their aim being to ascertain degree of similarity between laboratory and field systems. Microcosm dynamics, specifically diatom dynamics, most closely paralleled that found in reservoirs during late spring and early summer, a time of thermal stratification. During winter months when thermal stratification was absent or less pronounced, microcosm diatom populations diverged significantly from reservoir populations within 24 days. It is inferred that microcosm design and operating conditions have a major bearing on microcosm usefulness for environmental assessment.     

An intact soil-core microcosm method to evaluate the survival and vertical dispersal of Trichoderma atroviride SC1

Aim:  To develop an intact soil-core microcosm method to study the survival and vertical dispersal of an experimental biocontrol agent ( Trichoderma atroviride SC1) applied to the soil surface.
Methods and Results:  The soil for the microcosms was collected using iron pipes with perforations corresponding to different soil layers. The tool was inserted into the soil and gently removed with the soil core inside. Trichoderma atroviride SC1 was mixed with the top layer of soil in the pipe. The experiment was performed in 2006 and 2007, and data from the microcosms were compared with results obtained under field conditions in the locations in which, the microcosms were collected, in the same periods. The concentrations of T. atroviride SC1 in the soil were estimated immediately after treatment, and 1, 5, 9 and 18 weeks after treatment at both the soil surface and the above-mentioned depths. The development of T. atroviride SC1 populations in the microcosms during the 18 weeks of monitoring was similar to that observed under field conditions. The dispersal of conidia was affected by the application of water to the soil.
Conclusions:  Results demonstrate that this microcosm prototype can be used to model the behaviour of T. atroviride SC1 in soil.
Significance and Impact of the Study:  The intact soil-core microcosm is a reliable, easy-to-use, fast and cheap method that could also be used in studies of similar filamentous fungi to study their probable fate in the soil prior to their being introduced into the environment.     

In situ 15N-N2O site preference and O2 concentration dynamics disclose the complexity of N2O production processes in agricultural soil

Arable soil continues to be the dominant anthropogenic source of nitrous oxide (N₂O) emissions owing to application of nitrogen (N) fertilizers and manures across the world. Using laboratory and in situ studies to elucidate the key factors controlling soil N₂O emissions remains challenging due to the potential importance of multiple complex processes. We examined soil surface N₂O fluxes in an arable soil, combined with in situ high-frequency measurements of soil matrix oxygen (O₂) and N₂O concentrations, in situ ¹⁵N labeling, and N₂O ¹⁵N site preference (SP). The in situ O₂ concentration and further microcosm visualized spatiotemporal distribution of O₂ both suggested that O₂ dynamics were the proximal determining factor to matrix N₂O concentration and fluxes due to quick O₂ depletion after N fertilization. Further SP analysis and in situ ¹⁵N labeling experiment revealed that the main source for N₂O emissions was bacterial denitrification during the hot-wet summer with lower soil O₂ concentration, while nitrification or fungal denitrification contributed about 50.0% to total emissions during the cold-dry winter with higher soil O₂ concentration. The robust positive correlation between O₂ concentration and SP values underpinned that the O₂ dynamics were the key factor to differentiate the composite processes of N₂O production in in situ structured soil. Our findings deciphered the complexity of N₂O production processes in real field conditions, and suggest that O₂ dynamics rather than stimulation of functional gene abundances play a key role in controlling soil N₂O production processes in undisturbed structure soils. Our results help to develop targeted N₂O mitigation measures and to improve process models for constraining global N₂O budget.     

Effects of N-fertilisation on CH4 oxidation and production, and consequences for CH4 emissions from microcosms and rice fields

The world's growing human population causes an increasing demand for food, of which rice is one of the most important sources. In rice production nitrogen is often a limiting factor. As a consequence increasing amounts of fertiliser will have to be applied to maximise yields. There is an ongoing discussion on the possible effects of fertilisation on CH₄ emissions. We therefore investigated the effects of N‐fertiliser (urea) on CH₄ emission, production and oxidation in rice microcosms and field experiments. In the microcosms, a substantial but short‐lived reduction of CH₄ emission was observed after N‐addition to 43‐d‐old rice plants. Methane oxidation increased by 45%, demonstrated with inhibitor measurements and model calculations based on stable carbon isotope data (δ¹³CH₄). A second fertilisation applied to 92‐d‐old plants had no effect on CH₄ emission rates. The positive effect of additional N on methanotrophic bacteria was also found in vitro for potential CH₄ oxidation rates in soil and root samples from the microcosm and field experiments, indicated by elevated initial oxidation rates and reduced lag‐phases. Fertilisation did not affect methane production in the microcosms. In the field, the effects were diverse: methane production was inhibited in the topsoil, but stimulated instead in the bulk soil. Stimulation occurred probably in the anaerobic food chain at the level of hydrolytic or fermenting bacteria, because acetate, a methanogenic precursor, increased simultaneously. Combining field, microcosm and laboratory experiments we conclude that any agricultural treatment improving the N‐supply to the rice plants will also be favourable for the CH₄ oxidising bacteria. However, N‐fertilisation had only a transient influence and was counter‐balanced in the field by an elevated CH₄ production. A negative effect of the fertilisation was a transient increase of N₂O emissions from the microcosms. However, integrating over the season the global warming potential (GWP) of N₂O emitted after fertilisation was still negligible compared to the GWP of emitted CH₄.     

Contamination of Floodplain Soils along the Wupper River,Germany, with As,Co, Cu,Ni, Sb,and Zn and the Impact of Pre-definite Redox Variations on the Mobility of These Elements

This study demonstrates that floodplain soils of the River Wupper, Germany, are seriously contaminated with metal(loid)s. We used an automated biogeochemical microcosm system allowing controlled variation of redox potential (E_H) to assess the impact of pre-definite redox conditions on the dynamics of arsenic (As), cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), antimony (Sb), and zinc (Zn) in homogenized soil material taken from an acid floodplain soil. The concentrations of Co, Cu, Mn, Ni, Sb, and Zn in soil solution were low at low E_H, possibly due to the precipitation of metal sulfides, and increased with rising E_H, presumably caused by their association with dissolved organic carbon (DOC). A significant positive correlation between metal/DOC-ratio and E_H indicated that the binding of the metals to DOC shifted from stronger to weaker when E_H rose. Decreasing As concentrations with increasing E_H in soil solution indicated co-precipitation with Fe(hydr)oxides and/or oxidation of more soluble As(III) to less soluble As(V) during oxidation. The other studied elements seemed not to co-precipitate with newly formed Fe(hydr)oxides when E_H rose, possibly due to the prevailing low pH. In the future, the specific role of DOC and sulfur chemistry on metal(loid) dynamics should be elucidated more fully, and similar studies should be conducted with additional frequently flooded soils worldwide to verify these results.     

Acetylene reduction byDaviesia mimosoides under Eucalyptus

Summary Daviesia mimosoides is a common understorey legume in Eucalyptus forests of the Brindabella Range in southeastern Australia, capable of fixing atmospheric nitrogen. Rates of N fixation were measured by the acetylene-reduction technique over a growing season in the field. Pot trials under controlled conditions were also carried out to elucidate effects of soil moisture, temperature, and light. Average rates in the field varied from about 1–5 μ mol C₂H₄/g/h (wet weight of nodule), but rates up to 14 μ mol C₂H₄/g/h were measured in optimum controlled conditions. Annual N-fixation rates approximate 4.5–7.0 kg/ha. In pot trials, rate of acetylene reduction decreased with soil moisture to about−10 MPa tension, with a marked depression at about−6 MPa, but within the normal field range of soil moisture there was little correlation of moisture with average acetylene reduction rate. Rates were similar in the temperature range of 20–30°C, but were depressed by either low or high temperature (<10 or >30°C). Diurnal fluctuations in acetylene reduction rates were not correlated with solar radiation, but rates were limited by high mid-day temperatures.     

Plants Mediate the Sensitivity of Soil Respiration to Rainfall Variability

Soil respiration from grasslands plays a critical role in determining carbon dioxide (CO₂) feedbacks between soils and the atmosphere. In these often mesic systems, soil moisture and temperature tend to co-regulate soil respiration. Increasing variance of rainfall patterns may alter aboveground–belowground interactions and have important implications for the sensitivity of soil respiration to fluctuations in moisture and temperature. We conducted a set of field experiments to evaluate the independent and interactive effects of rainfall variability and plant–soil processes on respiration dynamics. Plant removal had strong effects on grassland soils, which included altered CO₂ flux owing to absence of root respiration; increased soil moisture and temperature; and reduced availability of dissolved organic carbon (DOC) for heterotrophic respiration by microorganisms. These plant-mediated effects interacted with our rainfall variability treatments to determine the sensitivity of soil respiration to both moisture and temperature. Using time-series multiple regression, we found that plants dampened the sensitivity of respiration to moisture under high variability rainfall treatments, which may reflect the relative stability of root contributions to total soil respiration. In contrast, plants increased the sensitivity of respiration to temperature under low variability rainfall treatment suggesting that the environmental controls on soil CO₂ dynamics in mesic habitats may be context dependent. Our results provide insight into the aboveground–belowground mechanisms controlling respiration in grasslands under variable rainfall regimes, which may be important for predicting CO₂ dynamics under current and future climate scenarios.     

Dynamics of microbial community during bioremediation of phenanthrene and chromium(VI)-contaminated soil microcosms

The combined effect of phenanthrene and Cr(VI) on soil microbial activity, community composition and on the efficiency of bioremediation processes has been studied. Biometer flask systems and soil microcosm systems contaminated with 2,000 mg of phenanthrene per kg of dry soil and different Cr(VI) concentrations were investigated. Temperature, soil moisture and oxygen availability were controlled to support bioremediation. Cr(VI) inhibited the phenanthrene mineralization (CO₂ production) and cultivable PAH degrading bacteria at levels of 500–2,600 mg kg⁻¹. In the bioremediation experiments in soil microcosms the degradation of phenanthrene, the dehydrogenase activity and the increase in PAH degrading bacteria counts were retarded by the presence of Cr(VI) at all studied concentrations (25, 50 and 100 mg kg⁻¹). These negative effects did not show a correlation with Cr(VI) concentration. Whereas the presence of Cr(VI) had a negative effect on the phenanthrene elimination rate, co-contamination with phenanthrene reduced the residual Cr(VI) concentration in the water exchangeable Cr(VI) fraction (WEF) in comparison with the soil microcosm contaminated only with Cr(VI). Clear differences were found between the denaturing gradient gel electrophoresis (DGGE) patterns of each soil microcosm, showing that the presence of different Cr(VI) concentrations did modulate the community response to phenanthrene and caused perdurable changes in the structure of the microbial soil community.     

Validation of Microcosms for Examining the Survival of Pseudomonas aureofaciens (lacZY) in Soil

Evaluating the safety and efficacy of a recombinant bacterium prior to its release into the terrestrial environment requires that risk assessment data be collected in the laboratory. Much of this information is obtained with the use of microcosms. The design of the microcosm significantly affects the ability of the recombinant microorganism to survive in soil and, thus, complicates the risk assessment process. To standardize microcosms for future use, we evaluated the survival of Pseudomonas aureofaciens 3732 RN-L11 (lacZY Rif(supr) Nal(supr)) in intact soil cores (5.0 by 15 cm; polyvinyl chloride core) and disturbed soil microcosms (50 g of fresh, sieved soil). Survival data were compared with those obtained during a field release. The intact soil core microcosm was shown to closely simulate results obtained in the field. The intact soil core microcosm closely predicts survival in bulk soil and in the rhizosphere of wheat. Data obtained with the microcosm were also similar when evaluated in separate studies in two different years. In 1993, P. aureofaciens survived for approximately 63 days in bulk soil and 96 days in the rhizosphere. The disturbed soil microcosm exhibited a much more rapid decline in population size (34 days to zero) than the intact core microcosm. We speculate that drying and sieving of soil for the disturbed soil microcosm affected the ability of the soil to support the survival of P. aureofaciens. These results demonstrate that a small, inexpensive, and simple intact soil core microcosm may be appropriate for risk assessment.     

Soil moisture surpasses elevated CO2 and temperature as a control on soil carbon dynamics in a multi-factor climate change experiment

Some single-factor experiments suggest that elevated CO₂ concentrations can increase soil carbon, but few experiments have examined the effects of interacting environmental factors on soil carbon dynamics. We undertook studies of soil carbon and nitrogen in a multi-factor (CO₂ × temperature × soil moisture) climate change experiment on a constructed old-field ecosystem. After four growing seasons, elevated CO₂ had no measurable effect on carbon and nitrogen concentrations in whole soil, particulate organic matter (POM), and mineral-associated organic matter (MOM). Analysis of stable carbon isotopes, under elevated CO₂, indicated between 14 and 19% new soil carbon under two different watering treatments with as much as 48% new carbon in POM. Despite significant belowground inputs of new organic matter, soil carbon concentrations and stocks in POM declined over four years under soil moisture conditions that corresponded to prevailing precipitation inputs (1,300 mm yr^?1). Changes over time in soil carbon and nitrogen under a drought treatment (approximately 20% lower soil water content) were not statistically significant. Reduced soil moisture lowered soil CO₂ efflux and slowed soil carbon cycling in the POM pool. In this experiment, soil moisture (produced by different watering treatments) was more important than elevated CO₂ and temperature as a control on soil carbon dynamics.     

Microbial and Root Components of Respiration of Sod-Podzolic Soils in Boreal Forest

The microbial contribution to the respiration of sod-podzolic soils has been estimated during two seasons (2012–2013) in boreal forest (Valdai district in Novgorod oblast, Russia) by a combination of methods of substrate induced respiration (SIR) and integration of components (IC). Despite the higher accuracy of SIR in estimating soil microbial respiration (R_mic), it is found that the combined application of these two methods results in a better correspondence of field experiments to the published data based on laboratory experiments. The contribution of microbial respiration differs between wooded and degraded sites. Hence, these sites should be investigated separately in upscaling studies of microbial respiration in soils of a boreal forest. The underestimation of microbial respiration should also be noted when using the IC method in field experiments. Among the main controls of R_mic are abiotic ones (soil temperature at a depth of 10 cm; month of the vegetation season), as well as the type of the mesohabitat. The seasonal dynamics of microbial respiration was related to the Selyaninov hydrothermal factor. Despite seasonal and cross-habitat differences in R_mic, it was specific for the particular type of soil and ecosystem.     

Advancement in soil microcosm apparatus for biogeochemical research

Application of soil microcosms has largely improved our understanding in biogeochemical processes, because all major environmental factors can be independently controlled. Recent advancement to improve the performance of soil microcosm has been made. The modifications include using a different incubation vessel and cap, replacing a magnetic stirrer with an overhead stirrer, providing temperature control for the microcosm, using data logger for continuous measurements of redox potential (Eh), pH and temperature, and applying automatic gas analysis. The modifications can be made in any combination to suit an individual's needs and budget.     

Short-Term Wavelike Dynamics of Bacterial Populations in Response to Nutrient Input from Fresh Plant Residues

The objectives of the research were to investigate short-term dynamics of bacterial populations in soil after a disturbance in the form of fresh organic matter incorporation and to investigate how these dynamics are linked to those of some environmental parameters. To reach these objectives, soil bacterial populations, mineral nitrogen, pH, and redox potential (ROP) were monitored daily for 1 month after incorporation of clover-grass (CG) plant material in microcosm experiments. Colony-forming units (CFUs) and direct microscopic counts of FDA-stained and FTTC-stained bacteria increased immediately after incorporation of the plant material, dropped within 2 days, and fluctuated thereafter. Harmonics analysis demonstrated that there were significant wavelike fluctuations with three or four significant peaks within 1 month after incorporation of clover-grass material. Peaks in CFUs were 1–2 days ahead of those in direct counts. Ammonium (NH₄) concentrations increased from the start of the experiments until nitrification commenced. Nitrate (NO₃) concentrations dropped immediately after plant incorporation, and then rose monotonically until the end of the experiments. There were no wavelike fluctuations in NH₄ and NO₃ concentrations, so that bacterial fluctuations could not be attributed to alternating mineral N shortages and sufficiencies. pH levels rose and declined with NH₄ levels. ROP dropped shortly before NH₄ concentrations rose, and increased before NH₄ concentrations decreased; there were no regular fluctuations in ROP, so that temporary oxygen shortages may not have been responsible for the observed fluctuations in bacterial populations. Thus, for the first time, regular wavelike dynamics were demonstrated for bacterial populations after perturbation by addition of fresh organic matter to soil, and several potential reasons for the death phase of the fluctuations could be excluded from further consideration.     

Ocean acidification and rising temperatures may increase biofilm primary productivity but decrease grazer consumption

Climate change may cause ecosystems to become trophically restructured as a result of primary producers and consumers responding differently to increasing CO₂ and temperature. This study used an integrative approach using a controlled microcosm experiment to investigate the combined effects of CO₂ and temperature on key components of the intertidal system in the UK, biofilms and their consumers (Littorina littorea). In addition, to identify whether pre-exposure to experimental conditions can alter experimental outcomes we explicitly tested for differential effects on L. littorea pre-exposed to experimental conditions for two weeks and five months. In contrast to predictions based on metabolic theory, the combination of elevated temperature and CO₂ over a five-week period caused a decrease in the amount of primary productivity consumed by grazers, while the abundance of biofilms increased. However, long-term pre-exposure to experimental conditions (five months) altered this effect, with grazing rates in these animals being greater than in animals exposed only for two weeks. We suggest that the structure of future ecosystems may not be predictable using short-term laboratory experiments alone owing to potentially confounding effects of exposure time and effects of being held in an artificial environment over prolonged time periods. A combination of laboratory (physiology responses) and large, long-term experiments (ecosystem responses) may therefore be necessary to adequately predict the complex and interactive effects of climate change as organisms may acclimate to conditions over the longer term.     

Responses of Antarctic soil microbial communities and associated functions to temperature and freeze-thaw cycle frequency

Climatic changes will not only result in higher overall temperature, but also in greater variability in weather conditions. Antarctic soils are subjected to extremely variable conditions in the form of frequent freeze–thaw cycles (FTCs), but the importance of alteration in FTC frequency, compared with increases in average temperature and indirect vegetation-mediated effects on soil microorganisms, is still unknown. We therefore designed two complementary microcosm experiments using undisturbed soil cores from Signy Island (60°43'S, 45°38'W) in the maritime Antarctic. The experiments consisted of soil core incubations with or without the overlying vegetation at four different temperatures and six different FTC regimes. We assessed bacterial and fungal density and community structure, as well as the density of several key genes in microbial nutrient cycles using a combination of RNA- and DNA-based molecular fingerprinting and quantitative PCR approaches in addition to enzymatic activity assays. Results showed that bacteria were more affected by warming than by changes in FTC frequency. In contrast, fungal community structure and abundance were mostly influenced by FTC frequency, as well as the presence of vegetation cover. The relative densities of several bacterial gene families involved in key steps of the N-cycle were affected by FTCs, while warming had little or no effect. The FTCs and incubation temperature also strongly influenced laccase enzymatic activity in soil. In total, our results suggest that, in addition to climatic warming, increased climatic variability may also have a profound impact on Antarctic microbial communities. Although these effects are difficult to detect with assays of total bacterial community structure, they do become manifest in the analysis of key functional gene densities.