The Effect of Interaction Between White-rot Fungi and Indigenous Microorganisms on Degradation of Polycyclic Aromatic Hydrocarbons in Soil

White-rot fungi applied for soil bioremediation have to compete with indigenous soil microorganisms. The effect of competition on both indigenous soil microflora and white-rot fungi was evaluated with regard to degradation of polycyclic aromatic hydrocarbons (PAH) with different persistence in soil. Sterile and non-sterile soil was artificially contaminated with ¹⁴C-labeled PAH consisting of three (anthracene), four (pyrene, benz[a]anthracene) and five fused aromatic rings (benzo[a]pyrene, dibenz[a,h]anthracene). The two fungi tested,Dichomitus squalens and Pleurotus ostreatus, produced similar amounts of ligninolytic enzymes in soil, but PAH mineralization by P. ostreatus was significantly higher. Compared to the indigenous soil microflora, P.ostreatus mineralized 5-ring PAH to a larger extent, while the indigenous microflora was superior in mineralizing 3-ring and 4-ring PAH. In coculture the special capabilities of both soil microflora and P. ostreatus were partly restricted due to antagonistic interactions, but essentially preserved. Thus, soil inoculation with P. ostreatus significantly increased the mineralization of high-molecular-weight PAH, and at the same time reduced the mineralization of anthracene and pyrene. Regarding the mineralization of low-molecular-weight PAH, the stimulation of indigenous soil microorganisms by straw amendment was more efficient than application of white-rot fungi.     

Fungal Degradation of the Bioplastic PHB (Poly-3-hydroxy- butyric acid)

PHB (poly-3-hydroxybutyric acid) is a thermoplastic polyester synthesized by Ralstonia eutropha and other bacteria as a form of intracellular carbon and energy storage and accumulated as inclusions in the cytoplasm of these bacteria. The degradation of PHB by fungi from samples collected from various environments was studied. PHB depolymerization was tested in vials containing a PHB-containing medium which were inoculated with isolates from the samples. The degradation activity was detected by the formation of a clear zone below and around the fungal colony. In total, 105 fungi were isolated from 15 natural habitats and 8 lichens, among which 41 strains showed PHB degradation. Most of these were deuteromycetes (fungi imperfecti) resembling species of Penicillium and Aspergillus and were isolated mostly from soils, compost, hay, and lichens. Soil-containing environments were the habitats from which the largest number of fungal PHB degraders were found. Other organisms involved in PHB degradation were observed. A total number of 31 bacterial strains out of 67 isolates showed clear zones on assay medium. Protozoa, possible PHB degraders, were also found in several samples such as pond, soil, hay, horse dung, and lichen. Lichen, a fungi and algae symbiosis, was an unexpected sample from which fungal and bacterial PHB degraders were isolated.     

Factors affecting polycyclic aromatic hydrocarbon biodegradation by Aspergillus flavus

This study investigated the ability of fungi isolated from highly contaminated soil to biodegrade polycyclic aromatic hydrocarbon (PAH) compounds, as well as the effect of several parameters on the biodegradation ability of these fungi. The isolated fungi were identified using ITS rDNA sequencing and tested using 2,6‐dichlorophinolendophenol to determine their preliminary ability to degrade crude oil. The top‐performing fungi, Aspergillus flavus and Aspergillus fumigatus, were selected to test their ability to biodegrade PAH compounds as single isolates. After 15 days of incubation, A. flavus degraded 82.7% of the total PAH compounds, with the complete degradation of six compounds, whereas A. fumigatus degraded 68.9% of the total PAHs, with four aromatic compounds completely degraded. We also tested whether different temperatures, pH, and nitrogen sources influenced the growth of A. flavus and the degradation rate. The degradation process was optimal at a temperature of 30°C, pH of 5.5, and with nitrogen in the form of yeast extract. Finally, the ability of the fungal candidate, A. flavus, to degrade PAH compounds under these optimum conditions was studied. The results showed that 95.87% of the total PAHs, including 11 aromatic compounds, were completely degraded after 15 days of incubation. This suggests that A. flavus is a potential microorganism for the degradation of PAH compounds in aqueous cultures.     

Impact of Land Use on Soluble Organic Nitrogen in Soil

Although it has been hypothesized that soluble organic nitrogen (SON) plays a central role in regulating productivity in some terrestrial ecosystems, the factors controlling the size of the SON pool in soil remain poorly understood. Therefore our principal aim in this work was to assess the impact of seven different land use systems (rough and managed grassland, deciduous and coniferous woodland, heathland, wetland and tilled land) on the size of the SON and inorganic N (NO ₃ ^– , NH ₄ ⁺ ) pools in the surface soil layer (0–15 cm). After extraction with deionised water, we found that in most cases the size of the water extractable organic N (WEON) pool was similar in size to the inorganic N pool. In contrast, the KCl extractable organic N (KClEON) pool constituted the dominant form of soluble N in soils under all land uses, perhaps indicating that significant amounts were held on the soil exchange phase. In contrast to inorganic N, which varied significantly with land use, the size of the KClEON and WEON pool was similar for all land uses with the exception of KClEON in tilled land, where significantly lower amounts were observed. We conclude that SON constitutes an important soil N pool in a broad range of land uses, and that its role in microbial N assimilation, plant nutrition and ecosystem responses to atmospheric N deposition warrants further attention. SAFRD, University of Newcastle, Newcastle-upon-Tyne, NE1 7RU, U.K.     

Linking field experiments to long-term simulation of impacts of nitrogen deposition on heathlands and moorlands

The results from three long-term field manipulation studies of the impacts of increased nitrogen deposition (0–120 kg N ha^–1 yr^–1) on lowland and upland heathlands in the UK were compared, to test if common responses are observed. Consistent increases in Calluna foliar N content and decreases in litter C:N ratios were found across all sites, while increases in N leaching were not observed at any site over the range 0–80 kg ha^–1 yr^–1. However, the response of Calluna biomass did vary between sites, possibly reflecting site differences in nutrient status and management histories. Five versions of a simulation model of heathland responses to N were developed, each reflecting different assumptions about the fate and turnover of soil N. Model outputs supported the deduction from mass balance calculations at two of the field sites that N additions have resulted in an increase in immobilisation; the latter was needed to prevent the model overestimating measured N leaching. However, this version of the model significantly underestimated Calluna biomass. Model versions, which included uptake of organic N by Callunaand re-mobilisation of N from the soil organic store provided some improvement in the fit between modelled and field biomass data, but re-mobilisation also led to an overestimation of N leaching. Quantification of these processes and their response to increased N deposition are therefore critical to interpreting experimental data and predicting the long-term impacts of atmospheric deposition on heathlands and moorlands.     

Impact of land use on soluble organic nitrogen in soil

Although it has been hypothesized that soluble organic nitrogen (SON) plays a central role in regulating productivity in some terrestrial ecosystems, the factors controlling the size of the SON pool in soil remain poorly understood. Therefore our principal aim in this work was to assess the impact of seven different land use systems (rough and managed grassland, deciduous and coniferous woodland, heathland, wetland and tilled land) on the size of the SON and inorganic N (NO^? ₃, NH⁺ ₄) pools in the surface soil layer (0–15 cm). After extraction with deionised water, we found that in most cases the size of the water extractable organic N (WEON) pool was similar in size to the inorganic N pool. In contrast, the KCl extractable organic N (KClEON) pool constituted the dominant form of soluble N in soils under all land uses, perhaps indicating that significant amounts were held on the soil exchange phase. In contrast to inorganic N, which varied significantly with land use, the size of the KClEON and WEON pool was similar for all land uses with the exception of KClEON in tilled land, where significantly lower amounts were observed. We conclude that SON constitutes an important soil N pool in a broad range of land uses, and that its role in microbial N assimilation, plant nutrition and ecosystem responses to atmospheric N deposition warrants further attention.     

Long-Term Maintenance of Rapid Atrazine Degradation in Soils Inoculated with Atrazine Degraders

Two different microbial communities able to degrade atrazine (atz) were inoculated in four different soils. The most critical factor affecting the success of inoculation was the soil pH and its organic matter (OM) content. In two alkaline soils (pH > 7), some inoculations led immediately to a strong increase of the biodegradation rate. In a third slightly acidic soil (pH = 6.1), only one inoculum could enhance atz degradation. In a soil amended with organic matter and straw (pH = 5.7, OM = 16.5%), inoculation had only little effect on atz dissipation on the short as well as on the long-term. Nine months after the microflora inoculations, atz was added again and rapid biodegradation in all alkaline inoculated soils was recorded, indicating the long-term efficiency of inoculation. In these soils, the number of atz degraders was estimated at between 6.5 × 10³ and 1.5 × 10⁶ (g of soil)^-1, using the most probable number (MPN) method. Furthermore, the presence of the atz degraders was confirmed by the detection of the gene atzA in these soils. Denaturing gradient gel electrophoresis (DGGE) analysis of the 16S rDNA genes indicated that the inoculated bacterial communities had little effect on the patterns of the indigenous soil microflora.     

Production and Consumption of NO in Forest and Pasture Soils from the Amazon Basin

NO production and the rate constant of NO consumptionin soil samples from Amazonian forest and pasturesites were determined in the laboratory. The purposewas to study NO production and consumption in soilsfrom both types of land use as functions of soiltemperature and soil moisture. NO productionincreased exponentially with soil temperature. Thedegree of increase depended on soil moisture,indicating that the response of NO production to achange in soil temperature is most pronounced at acertain intermediate soil moisture. NO productionpeaked at a soil moisture of 0.10 g g^-1, correspondingto 0.27 and 0.38 water-filled pore space for forestand pasture soils, respectively. The optimum soilmoisture for which maximum NO release was observedwas independent of soil temperature. NO consumptionwas most efficient at high soil temperatures andunder dry soil conditions. NO release wasapproximately 10 times larger for forest than pasturesoil. The difference may be related to the higherNO₃ ^- content in forest soil.     

Changes in Extractability of Cr and Pb in a Polycontaminated Soil After Bioaugmentation With Microbial Producers of Biosurfactants, Organic Acids and Siderophores

Partly because of the low bioavailability of metals, the soil cleaning-up using phytoremediation is usually time-consuming. In order to enhance the amount of metals at the plant's disposal, the soil bioaugmentation coupled together with phytoextraction is an emerging technology. In this preliminary work, two agricultural soils which mainly differed in their Cr, Hg and Pb contents (LC, low-contaminated soil; HC, high-contaminated soil) were bioaugmented in laboratory conditions by either bacterial (Bacillus subtilis, Pseudomonas aeruginosa, Pseudomonas fluorescens or Ralstonia metallidurans) or fungal inocula (Aspergillus niger or Penicillium simplicissimum) and incubated during three weeks. The LC soil pots bioaugmented with A. niger and P. aeruginosa contained higher concentrations of Cr (0.08 and 0.25 mg.kg⁻¹ dw soil) and Pb (0.25 and 0.3 mg.kg⁻¹ dw soil) in the exchangeable fraction F1 (extraction with MgCl₂) by comparison with the non-bioaugmented soil where neither Cr nor Pb was detected. Conversely, immobilization of Cr and Pb in the soil were observed with the other microorganisms. The soil bioaugmentation not only modified the metal speciation for the most easily extractable fractions but also modified the distribution of metals in the other fractions, to a lesser extent nevertheless. The difference in microbial concentrations between the bioaugmented or not HC soils reached up to 1.8 log units. Thus the microorganisms that we chose for the soil bioaugmentation were competitive towards the indigenous microflora. The PCA analysis showed close positive relationships between the microorganisms which potentially produced siderophores in the soil and the amount of Cr and Pb in the fraction F1.     

Effects of Acidification and its Mitigation with Lime and Wood Ash on Forest Soil Processes in Southern Sweden. A Joint Multidisciplinary Study

A joint multidisciplinary investigation was undertaken to studythe effects of lime and wood ash applications on two Norway spruce forest Spodosolic soils. The two sites, typical for southern Sweden, were treated in 1994 with either 3.25 t ha^-1 dolomite or 4.28 t ha^-1 wood ash (Horröd site) or in 1984 with either 3.45 or 8.75 t ha^-1 dolomite (Hasslöv site). Both sites show signs of acidification by atmospheric anthropogenic deposition and possessed low soil pH(4.3) and high concentrations of inorganic Al (35 M) in theupper illuvial soil solution. The prevailing soil conditions indicated perturbed soil processes. Following treatment with lime or wood ash, the soil conditions were dramatically altered. Cation exchange capacity (CEC) and base saturation (BS) was considerable increased after addition. Four years after application most of the added Ca and Mg was still present in the mor layer. Fifteen years after application,Mg in particular, became integrated deeper in the soil profile with a greater proportion lost by leaching incomparison to Ca. The concentrations of these ions were greatestin the mor layer soil solutions and Mg had higher mobility givinghigher concentrations also deeper in the profile. Four years after treatment, the application of wood ash and limeresulted in lower pH values and higher inorganic Al in mineral subsoil solutions compared to the untreated soil. We hypothesize that this was probably due to an increased flow of hydrogen ionsfrom the upper soil as a result of displacement by Ca and Mg ionsin the enlarged exchangeable pool. In contrast, fifteen years after lime and wood ash application, the mineral subsoil horizonspossessed a higher pH and lower soil solution Al content than theuntreated plots.Liming promoted soil microbial activity increasing soil respiration 10 to 36%. This is in the same range as net carbon exchange for forests in northern Sweden and could potentially have a climatological impact. The turnover of low molecularweight organic acids (LMWOA) by the soil microbial biomass werecalculated to contribute 6 to 20% to this CO₂ evolution.At Horröd, citrate and fumarate were the predominant LMWOAs with lowest concentrations found in the treated areas. In contrast, at the Hasslöv site, propionate and malonate were the most abundant LMWOAs. Higher microbial activity in the upper soil horizons was also theprobable cause of the considerably higher DOC concentrations observed in the soil solution of ash and lime treated areas. Thelime-induced increase in DOC levels at Hasslöv could be attributed to increases in the 3–10 kDa hydrophobic size fraction. Liming also promoted nitrification with high liming doses leading to extreme concentrations of NO₃ ^- (1 mM) in soil solution.At Hasslöv the community of mycorrhizal fungi was dramatically changed by the addition of lime, with only four of 24 species recorded being common to both control and treated areas.Many of the observed effects of lime and ash treatment can be viewed as negative in terms of forest sustainability. After fouryears of treatment, there was a decrease in the pH of the soil solution and higher concentrations of inorganic Al and DOC. Increased organic matter turnover, nitrification and NO₃ ^-leakage were found at Hasslöv. Considering that the weathering rate and the mineral nutrient uptake by trees is mostprobably governed by mycorrhizal hyphae etchingmineral grains in the soil, it is important to maintain this ability of the mycorrhizal fungi. The lime and ash-induced changed mycorrhizal community structure may significantly affect this capability. In light of this investigation and others, as reviewed by Lundström et al. (2003), the implications ofliming on forest health are multifaceted with complex relationships occurring over both space and time.     

Use of fungal technology in soil remediation: A Case Study

Two white rot fungi Irpex lacteus and Pleurotus ostreatus and a PAH-degrading bacterial strain of Pseudomonas putida were used as inoculum for bioremediation of petroleum hydrocarbon-contaminated soil from a manufactured-gas-plant-area. Also two cocultures comprising a fungus with Pseudomonas putida were applied. After 10-week treatment out of 12 different PAHs, concentration of phenanthrene, anthracene, fluoranthene and pyrene decreased up to 66%. The ecotoxicity of the soil after bioremediation did not reveal any effect on the survival of Daphnia magna, a crustacian. However, the toxic effect on seed germination of plant Brassica alba and oxidoreductase activity of bacterium Bacillus cereus decreased after 5 and 10 weeks of treatment.     

Isolation and identification of potential fungal species for spent engine lubrication oil remediation in Peninsular Malaysia

Improper disposal and spills of spent engine oil into the environment can result in contamination, which eventually affects humans through the food chain. Mycoremediation is an effective and inexpensive alternative to clean up spent engine oil contamination. In recent work, the potential effectiveness of fungi for degrading spent engine oil was confirmed, with the species identified through molecular identification. Fungi that were able to grow in Bushnell Haas Broth supplied with spent engine oil were identified with the potential to utilize spent engine oil as a carbon source. Six species of fungi namely Penicillium simplicissimum, Aspergillus nidulans, Aspergillus niger, Trichoderma longibrachiatum, Aspergillus ustus, and Aspergillus flavus were successfully identified in this study. Over a course of seven days, P. simplicissimum (21.11 percent) was identified as the most effective fungi in degrading spent engine oil, followed by A. nidulans (17.75 percent), A. niger (15.85 percent), T. longibrachiatum (15.12 percent), A. ustus (15.02 percent), and A. flavus (11.80 percent). As these species of fungi were isolated from the natural environment in Peninsular Malaysia, the potential of using these fungi as mycoremediation of spent engine oil was therefore confirmed.     

Biosorption study of potential fungi for copper remediation from Peninsular Malaysia

The ability to grow in heavy metal contaminated areas and absorb heavy metals from the environment make fungi a potentially viable biological‐based technology for remediating hazardous heavy metals in soil. In this study, 10 fungi from a copper (Cu)‐polluted area in Malaysia were isolated, with the four highest growth fungi identified as Simplicillium subtropicum, Fusarium solani, Aspergillus tamari, and Aspergillus niger. Results from toxicity and biosorption testing showed that A. niger and F. solani grew the fastest in the presence of Cu, but exhibited lower Cu uptake per unit of biomoass. In contrast, A. tamarii and S. subtropicum had lower growth rates, but showed better uptake of Cu per unit of biomass. S. subtropicum was identified as the best species for bioremediation because it had the highest Cu uptake and positive growth measured in the presence of Cu at concentrations below 100 mg/L. A niger proved to be most suitable for bioremediation if the concentration of Cu exceeds 100 mg/L.     

Linking field experiments to long-term simulation of impacts of nitrogen deposition on heathlands and moorlands

The results from three long-term field manipulation studies of the impacts of increased nitrogen deposition (0–120 kg N ha^?1 yr^?1) on lowland and upland heathlands in the UK were compared, to test if common responses are observed. Consistent increases in Calluna foliar N content and decreases in litter C:N ratios were found across all sites, while increases in N leaching were not observed at any site over the range 0–80 kg ha^?1 yr^?1. However, the response of Calluna biomass did vary between sites, possibly reflecting site differences in nutrient status and management histories. Five versions of a simulation model of heathland responses to N were developed, each reflecting different assumptions about the fate and turnover of soil N. Model outputs supported the deduction from mass balance calculations at two of the field sites that N additions have resulted in an increase in immobilisation; the latter was needed to prevent the model overestimating measured N leaching. However, this version of the model significantly underestimated Calluna biomass. Model versions, which included uptake of organic N by Callunaand re-mobilisation of N from the soil organic store provided some improvement in the fit between modelled and field biomass data, but re-mobilisation also led to an overestimation of N leaching. Quantification of these processes and their response to increased N deposition are therefore critical to interpreting experimental data and predicting the long-term impacts of atmospheric deposition on heathlands and moorlands.     

Utilisation of Supernatants of Pure Cultures of Streptomyces thermocarboxydus NH50 to Reduce Chromium Toxicity and Mobility in Contaminated Soils

Chromium is a heavy metal used in various industrial sectors. Improper handling and storage of chromium-laden effluents or wastes can lead to the pollution of the environment. The most toxic form is the more mobile one: hexavalent chromium Cr(VI). The reduction of Cr(VI) results in the immobilisation of chromium into its less toxic trivalent form Cr(III). This phenomenon may prevent the contamination of groundwater when the soil in the vadose zone is contaminated. Many bacteria have been isolated from contaminated soils and described to reduce Cr(VI) into Cr(III). A new Cr(VI)-reducing strain, identified as a Streptomyces thermocarboxydus,has been isolated and studied in our laboratories for its ability to reduce Cr(VI). This aerobic bacterium, in contrast to other genera described which mediate reduction via enzymes, produces reducing agents into the culture supernatants. Cr(VI) reduction by these substances is accelerated by the presence of small concentration of cupric ions (Cu²⁺). The reducing agent(s) can be easily recovered from the bacterial cultures and used as cell-free solution to treat contaminated soils by an in situ or ex situ processes.     

Biodegradation of Electrospun Poly-(ɛ-caprolactone) Non-woven Fabrics by Pure-Cultured Soil Filamentous Fungi

The biodegradation of electrospun nano-fibers of poly(-caprolactone) (PCL) was initially investigated with respect to the environmental application of PCL non-woven fabrics, using pure-cultured soil filamentous fungi, Aspergillus oryzae, Penicillium caseicolum, P. citrinum, Mucor sp., Rhizopus sp., Curvularia sp., and Cladosporium sp. Three kinds of non-woven PCL fabrics with different mean fiber diameters (330, 360, and 510 nm) were prepared by changing the viscosities of the pre-spun PCL solutions (150, 210, and 310 cPs, respectively). All of the pure-line soil filamentous fungi tested grew on the two fiber materials. Electron microscopy was used to observe the biodegradation processes revealing remarkable growth of two fungi, Rhizopus sp. and Mucor sp., along with the accompanying collapse of the nano-fiber matrices. In the biochemical oxygen demand (BOD) test, the biodegradation of the 330 nm PCL nano-fibers by Rhizopus sp. and Mucor sp. exceeded 20 and 30% carbon dioxide generation, respectively. The biodegradability of the PCL non-woven fabrics decreased with the mean fiber diameter and the 330 nm PCL nano-fiber that was made from 150 cPs solution (concentration, 7 wt%) exhibited the highest biodegradability. These results might offer some clues for the applications of the PCL non-woven fabrics having the controlled biodegradability in the environmental uses.     

Effects of Nitrogen Deposition on Bryophyte Species Composition of Calcareous Grasslands

Regular additions of NH₄NO₃ (35–140 kg N ha⁻¹ yr⁻¹) and (NH₄)₂SO₄ (140 kg N ha⁻¹ yr⁻¹) to a calcareous grassland in northern England over a period of 12 years have resulted in a decline in the frequency of the indigenous bryophyte species and the establishment of non-indigenous calcifuge species, with implications for the structure and composition of this calcareous bryophyte community. The lowest NH₄NO₃ additions of 35 kg N ha⁻¹ yr⁻¹ produced significant declines in frequency of Hypnum cupressiforme, Campylium chrysophyllum, and Calliergon cuspidatum. Significant reductions in frequency at higher NH₄NO₃ application rates were recorded for Pseudoscleropodium purum, Ctenidum molluscum, and Dicranum scoparium. The highest NH₄NO₃ and (NH₄)₂SO₄ additions provided conditions conducive for the establishment of two typical calcifuges – Polytrichum spp. and Campylopus introflexus, respectively. Substrate-surface pH measurements showed a dose-related reduction in pH with increasing NH₄NO₃ deposition rates of 1.6 pH units between the control and highest deposition rate, and a further significant fall in pH, of >1 pH unit, between the NH₄NO₃ and (NH₄)₂SO₄ treatments. These results suggest that indigenous bryophyte composition may be at risk from nitrogen deposition rates of 35 kg N ha⁻¹ yr⁻¹ or less. These effects are of particular concern for rare or endangered species of low frequency.     

Trends in Water Chemistry in a Maple Forest on a Steep Slope

Year-to-year variation in SO₄ ^2-,NO₃ ^-, Ca²⁺, K⁺, and Mg²⁺concentrations in forest floor and mineral soil percolatefrom a forested, podzolic soil at the Turkey Lakes Watershedon the Precambrian Shield was assessed for monotonic trendsbetween 1986 and 1995. Our objective was to examine howrapidly ion concentrations in soil percolate equilibratedafter stabilization of SO₄ ^2- concentrations inprecipitation. Significant negative trends were detected inmonthly Ca²⁺, and Mg²⁺ concentrations in forestfloor and SO₄ ^2-, Ca²⁺, and Mg²⁺ inmineral soil percolate during the 10-year-period. Thedecline in Ca²⁺ and Mg²⁺ was greater than annualdecreases in SO₄ ^2- and NO₃ ^- in forestfloor percolate and proportional to the reduction inSO₄ ^2- in mineral soil percolate. Response ofmineral soil percolate to a 15 mol_c L^-1SO₄ ^2- decrease in wet-only precipitation between1985 and 1986 was a gradual decline in SO₄ ^2-concentration through 1995. The five-year meanSO₄ ^2- concentration in bulk precipitation, forestfloor percolate, and mineral soil percolate decreased 8, 9and 18 mol_c L^-1 from 1986–90 to 1991–95.Microbial (mineralization of organic S) and sorption(release from and/or retention in the pool of insolubleSO₄ ^2-) processes in the soil were logicalexplanations for the observed changes in SO₄ ^2- inmineral soil percolate.     

Metabolic and Environmental Control on Methane Emission from Soils: Mechanistic Studies of Mesotrophic Fen in West Siberia

The biological mechanisms regulating methane emission fromnatural wetlands are the focus of this article. A novel techniqueprovides estimates of the distribution of CH₄ sources withinan undisturbed soil profile by recording the transient gasdynamics after soil enclosure by deep (50 cm) chamber. Thecombined use of conventional surface and soil chambers across a200-m fen transect allowed us to relate the observed methaneemission to its instant generation and uptake. Surprisingly, themethane generation was relatively constant (9–12 mg CH₄-C hr^-1 m^-2) and highly variable net emission (0.2–20 mg CH₄-C hr^-1 m^-2) was closely correlated (r = –0.809)with methane uptake. In laboratory incubations, CH₄ uptakefollowed Michaelis-Menten kinetics. Added chloride and nitrateirrespective of the cation's nature suppressed uptake as a strongnoncompetitive inhibitors (K _i 0.5 mM). The methaneformation turned out to be unstable and under anaerobicincubation, the formation of CH₄, CO₂ andH₂displayed sustained weekly oscillations. We conclude that effectsof environmental factors alone are not sufficient topredict the variation in emission, which depends also on metabolic control of respective soil organisms. The multitude ofsuch controls is dependent on diversity of respective soilorganisms and could be grouped into a limited number ofcategories considerably simplifying large-scale simulations.