Atmospheric CH4 oxidation by Arctic permafrost and mineral cryosols as a function of water saturation and temperature

The response of methanotrophic bacteria capable of oxidizing atmospheric CH₄ to climate warming is poorly understood, especially for those present in Arctic mineral cryosols. The atmospheric CH₄ oxidation rates were measured in microcosms incubated at 4 °C and 10 °C along a 1‐m depth profile and over a range of water saturation conditions for mineral cryosols containing type I and type II methanotrophs from Axel Heiberg Island (AHI), Nunavut, Canada. The cryosols exhibited net consumption of ~2 ppmv CH₄ under all conditions, including during anaerobic incubations. Methane oxidation rates increased with temperature and decreased with increasing water saturation and depth, exhibiting the highest rates at 10 °C and 33% saturation at 5 cm depth (260 ± 60 pmol CH₄ gdw^?1 d^?1). Extrapolation of the CH₄ oxidation rates to the field yields net CH₄ uptake fluxes ranging from 11 to 73 μmol CH₄ m^?2 d^?1, which are comparable to field measurements. Stable isotope mass balance indicates ~50% of the oxidized CH₄ is incorporated into the biomass regardless of temperature or saturation. Future atmospheric CH₄ uptake rates at AHI with increasing temperatures will be determined by the interplay of increasing CH₄ oxidation rates vs. water saturation and the depth to the water table during summer thaw.     

Molecular Analyses of Novel Methanotrophic Communities in Forest Soil That Oxidize Atmospheric Methane

Forest and other upland soils are important sinks for atmospheric CH₄, consuming 20 to 60 Tg of CH₄ per year. Consumption of atmospheric CH₄ by soil is a microbiological process. However, little is known about the methanotrophic bacterial community in forest soils. We measured vertical profiles of atmospheric CH₄ oxidation rates in a German forest soil and characterized the methanotrophic populations by PCR and denaturing gradient gel electrophoresis (DGGE) with primer sets targeting the pmoA gene, coding for the α subunit of the particulate methane monooxygenase, and the small-subunit rRNA gene (SSU rDNA) of all life. The forest soil was a sink for atmospheric CH₄ in situ and in vitro at all times. In winter, atmospheric CH₄ was oxidized in a well-defined subsurface soil layer (6 to 14 cm deep), whereas in summer, the complete soil core was active (0 cm to 26 cm deep). The content of total extractable DNA was about 10-fold higher in summer than in winter. It decreased with soil depth (0 to 28 cm deep) from about 40 to 1 μg DNA per g (dry weight) of soil. The PCR product concentration of SSU rDNA of all life was constant both in winter and in summer. However, the PCR product concentration of pmoA changed with depth and season. pmoA was detected only in soil layers with active CH₄ oxidation, i.e., 6 to 16 cm deep in winter and throughout the soil core in summer. The same methanotrophic populations were present in winter and summer. Layers with high CH₄ consumption rates also exhibited more bands of pmoA in DGGE, indicating that high CH₄ oxidation activity was positively correlated with the number of methanotrophic populations present. The pmoA sequences derived from excised DGGE bands were only distantly related to those of known methanotrophs, indicating the existence of unknown methanotrophs involved in atmospheric CH₄ consumption.     

Methanotrophic community structure and activity under warming and grazing of alpine meadow on the Tibetan Plateau

Knowledge about methanotrophs and their activities is important to understand the microbial mediation of the greenhouse gas CH₄ under climate change and human activities in terrestrial ecosystems. The effects of simulated warming and sheep grazing on methanotrophic abundance, community composition, and activity were studied in an alpine meadow soil on the Tibetan Plateau. There was high abundance of methanotrophs (1.2–3.4 × 10⁸ pmoA gene copies per gram of dry weight soil) assessed by real-time PCR, and warming significantly increased the abundance regardless of grazing. A total of 64 methanotrophic operational taxonomic units (OTUs) were obtained from 1,439 clone sequences, of these OTUs; 63 OTUs (98.4%) belonged to type I methanotrophs, and only one OTU was Methylocystis of type II methanotrophs. The methanotroph community composition and diversity were not apparently affected by the treatments. Warming and grazing significantly enhanced the potential CH₄ oxidation activity. There were significantly negative correlations between methanotrophic abundance and soil moisture and between methanotrophic abundance and NH₄–N content. The study suggests that type I methanotrophs, as the dominance, may play a key role in CH₄ oxidation, and the alpine meadow has great potential to consume more CH₄ under future warmer and grazing conditions on the Tibetan Plateau.     

Effects of ammonium on the activity and community of methanotrophs in landfill biocover soils

The influence of NH₄⁺ on microbial CH₄ oxidation is still poorly understood in landfill cover soils. In this study, effects of NH₄⁺ addition on the activity and community structure of methanotrophs were investigated in waste biocover soil (WBS) treated by a series of NH₄⁺-N contents (0, 100, 300, 600 and 1200 mg kg⁻¹). The results showed that the addition of NH₄⁺-N ranging from 100 to 300 mg kg⁻¹ could stimulate CH₄ oxidation in the WBS samples at the first stage of activity, while the addition of an NH₄⁺-N content of 600 mg kg⁻¹ had an inhibitory effect on CH₄ oxidation in the first 4 days. The decrease of CH₄ oxidation rate observed in the last stage of activity could be caused by nitrogen limitation and/or exopolymeric substance accumulation. Type I methanotrophs Methylocaldum and Methylobacter, and type II methanotrophs (Methylocystis and Methylosinus) were abundant in the WBS samples. Of these, Methylocaldum was the main methanotroph in the original WBS. With incubation, a higher abundance of Methylobacter was observed in the treatments with NH₄⁺-N contents greater than 300 mg kg⁻¹, which suggested that NH₄⁺-N addition might lead to the dominance of Methylobacter in the WBS samples. Compared to type I methanotrophs, the abundance of type II methanotrophs Methylocystis and/or Methylosinus was lower in the original WBS sample. An increase in the abundance of Methylocystis and/or Methylosinus occurred in the last stage of activity, and was likely due to a nitrogen limitation condition. Redundancy analysis showed that NH₄⁺-N and the C/N ratio had a significant influence on the methanotrophic community in the WBS sample.     

Radioactive Fingerprinting of Microorganisms That Oxidize Atmospheric Methane in Different Soils

Microorganisms that oxidize atmospheric methane in soils were characterized by radioactive labelling with ¹⁴CH₄ followed by analysis of radiolabelled phospholipid ester-linked fatty acids (¹⁴C-PLFAs). The radioactive fingerprinting technique was used to compare active methanotrophs in soil samples from Greenland, Denmark, the United States, and Brazil. The ¹⁴C-PLFA fingerprints indicated that closely related methanotrophic bacteria were responsible for the oxidation of atmospheric methane in the soils. Significant amounts of labelled PLFAs produced by the unknown soil methanotrophs coeluted with a group of fatty acids that included i17:0, a17:0, and 17:1ω8c (up to 9.0% of the total ¹⁴C-PLFAs). These PLFAs are not known to be significant constituents of methanotrophic bacteria. The major PLFAs of the soil methanotrophs (73.5 to 89.0% of the total PLFAs) coeluted with 18:1 and 18:0 fatty acids (e.g., 18:1ω9, 18:1ω7, and 18:0). The ¹⁴C-PLFAs fingerprints of the soil methanotrophs that oxidized atmospheric methane did not change after long-term methane enrichment at 170 ppm CH₄. The ¹⁴C-PLFA fingerprints of the soil methanotrophs were different from the PLFA profiles of type I and type II methanotrophic bacteria described previously. Some similarity at the PLFA level was observed between the unknown soil methanotrophs and the PLFA phenotype of the type II methanotrophs. Methanotrophs in Arctic, temperate, and tropical regions assimilated between 20 and 54% of the atmospheric methane that was metabolized. The lowest relative assimilation (percent) was observed for methanotrophs in agricultural soil, whereas the highest assimilation was observed for methanotrophs in rain forest soil. The results suggest that methanotrophs with relatively high carbon conversion efficiencies and very similar PLFA compositions dominate atmospheric methane metabolism in different soils. The characteristics of the methane metabolism and the ¹⁴C-PLFA fingerprints excluded any significant role of autotrophic ammonia oxidizers in the metabolism of atmospheric methane.     

Methane oxidation and methane driven redox process during sequential reduction of a flooded soil ecosystem

A laboratory incubation study conducted to assess the temporal variation of CH₄ oxidation during soil reduction processes in a flooded soil ecosystem. A classical sequence of microbial terminal electron accepting process observed following NO₃ ^? reduction, Fe³⁺ reduction, SO₄ ^2? reduction and CH₄ production in flooded soil incubated under initial aerobic and helium-flushed anaerobic conditions. CH₄ oxidation in the slurries was influenced by microbial redox process during slurry reduction. Under aerobic headspace condition, CH₄ oxidation rate (k) was stimulated by 29 % during 5 days (NO₃ ^? reduction) and 32 % during both 10 days (Fe³⁺) and 20 days (early SO₄ ^2? reduction) over unreduced slurry. CH₄ oxidation was inhibited at the later methanogenic period. Contrastingly, CH₄ oxidation activity in anaerobic incubated slurries was characterized with prolonged lag phase and lower CH₄ oxidation. Higher CH₄ oxidation rate in aerobically incubated flooded soil was related to high abundance of methanotrophs (r?=?0.994, p?<?0.01) and ammonium oxidizers population (r?=?0.184, p?<?0.05). Effect of electron donors NH₄ ⁺, Fe^2+, S^2? on CH₄ oxidation assayed to define the interaction between reduced inorganic species and methane oxidation. The electron donors stimulated CH₄ oxidation as well as increased the abundance of methanotrophic microbial population except S^2? which inhibited the methanotrophic activity by affecting methane oxidizing bacterial population. Our result confirmed the complex interaction between methane-oxidizing microbial groups and redox species during sequential reduction processes of a flooded soil ecosystem.     

Field application of nitrogen and phenylacetylene to mitigate greenhouse gas emissions from landfill cover soils: effects on microbial community structure

Landfills are large sources of CH₄, but a considerable amount of CH₄ can be removed in situ by methanotrophs if their activity can be stimulated through the addition of nitrogen. Nitrogen can, however, lead to increased N₂O production. To examine the effects of nitrogen and a selective inhibitor on CH₄ oxidation and N₂O production in situ, 0.5 M of NH₄Cl and 0.25 M of KNO₃, with and without 0.01% (w/v) phenylacetylene, were applied to test plots at a landfill in Kalamazoo, MI from 2007 November to 2009 July. Nitrogen amendments stimulated N₂O production but had no effect on CH₄ oxidation. The addition of phenylacetylene stimulated CH₄ oxidation while reducing N₂O production. Methanotrophs possessing particulate methane monooxygenase and archaeal ammonia-oxidizers (AOAs) were abundant. The addition of nitrogen reduced methanotrophic diversity, particularly for type I methanotrophs. The simultaneous addition of phenylacetylene increased methanotrophic diversity and the presence of type I methanotrophs. Clone libraries of the archaeal amoA gene showed that the addition of nitrogen increased AOAs affiliated with Crenarchaeal group 1.1b, while they decreased with the simultaneous addition of phenylacetylene. These results suggest that the addition of phenylacetylene with nitrogen reduces N₂O production by selectively inhibiting AOAs and/or type II methanotrophs.     

Response and adaptation of different methanotrophic bacteria to low methane mixing ratios

Described genera of methanotrophic bacteria are present in most upland soils, but it is not known whether these are sufficiently oligotrophic to oxidize methane at its trace atmospheric mixing ratio of 1.75 ppmv. Members of the genera Methylocystis, Methylosinus, Methylocaldum and Methylobacter were isolated from different upland soils and compared with type strains for growth and activity under low methane mixing ratios. The specific affinity (a0s) varied by about one order of magnitude among different methanotrophs. It was highest in some Methylocystis spp., suggesting that these were the most oligotrophic. In direct tests, the threshold mixing ratio of methane required by most methanotrophs for growth ranged from 100 to greater than 1000 ppmv. However, two Methylocystis strains grew at only 10-100 ppmv of methane and one oxidized atmospheric methane for >3 months with little or no decline in the absolute rate. The results show that some cultivated methanotrophic bacteria are much more oligotrophic than others, and may contribute to atmospheric methane oxidation in soils. However, it is likely that these need additional energy sources for long-term survival, and that uncultivated groups of methanotrophic bacteria are primarily responsible for the process in soils possessing high methane oxidation rates.     

Effects of ammonium-based fertilisation on microbialprocesses involved in methane emission from soilsplanted with rice

The emission of the greenhouse gas CH₄ from ricepaddies is strongly influenced by management practicessuch as the input of ammonium-based fertilisers. Weassessed the impact of different levels (200 and 400kgN.ha^–1) of urea and (NH₄)₂HPO₄on the microbial processes involved in production andconsumption of CH₄ in rice field soil. We usedcompartmented microcosms which received fertilisertwice weekly. Potential CH₄ production rates weresubstantially higher in the rice rhizosphere than inunrooted soil, but were not affected by fertilisation.However, CH₄ emission was reduced by the additionof fertiliser and was negatively correlated with porewater NH ₄ ^plus concentration, probably as theconsequence of elevated CH₄ oxidation due tofertilisation. CH₄ oxidation as well as numbersof methanotrophs was distinctly stimulated by theaddition of fertiliser and by the presence of the riceplant. Without fertiliser addition,nitrogen-limitation of the methanotrophs will restrictthe consumption of CH₄. This may have a majorimpact on the global CH₄ budget, asnitrogen-limiting conditions will be the normalsituation in the rice rhizosphere. Elevated potentialnitrifying activities and numbers were only detectedin microcosms fertilised with urea. However, asubstantial part of the nitrification potential in therhizosphere of rice was attributed to the activity ofmethanotrophs, as was demonstrated using theinhibitors CH₃F and C₂H₂.     

Contribution of Methanotrophic and Nitrifying Bacteria to CH4 and NH4+ Oxidation in the Rhizosphere of Rice Plants as Determined by New Methods of Discrimination

Methanotrophic and nitrifying bacteria are both able to oxidize CH₄ as well as NH₄⁺. To date it is not possible to estimate the relative contribution of methanotrophs to nitrification and that of nitrifiers to CH₄ oxidation and thus to assess their roles in N and C cycling in soils and sediments. This study presents new options for discrimination between the activities of methanotrophs and nitrifiers, based on the competitive inhibitor CH₃F and on recovery after inhibition with C₂H₂. By using rice plant soil as a model system, it was possible to selectively inactivate methanotrophs in soil slurries at a CH₄/CH₃F/NH₄⁺ molar ratio of 0.1:1:18. This ratio of CH₃F to NH₄⁺ did not affect ammonia oxidation, but methane oxidation was inhibited completely. By using the same model system, it could be shown that after 24 h of exposure to C₂H₂ (1,000 parts per million volume), methanotrophs recovered within 24 h while nitrifiers stayed inactive for at least 3 days. This gave an “assay window” of 48 h when only methanotrophs were active. Applying both assays to model microcosms planted with rice plants demonstrated a major contribution of methanotrophs to nitrification in the rhizosphere, while the contribution of nitrifiers to CH₄ oxidation was insignificant.     

Linking activity,composition and seasonal dynamics of atmospheric methane oxidizers in a meadow soil

Microbial oxidation is the only biological sink for atmospheric methane. We assessed seasonal changes in atmospheric methane oxidation and the underlying methanotrophic communities in grassland near Giessen (Germany), along a soil moisture gradient. Soil samples were taken from the surface layer (0–10 cm) of three sites in August 2007, November 2007, February 2008 and May 2008. The sites showed seasonal differences in hydrological parameters. Net uptake rates varied seasonally between 0 and 70 μg CH₄ m⁻² h⁻¹. Greatest uptake rates coincided with lowest soil moisture in spring and summer. Over all sites and seasons, the methanotrophic communities were dominated by uncultivated methanotrophs. These formed a monophyletic cluster defined by the RA14, MHP and JR1 clades, referred to as upland soil cluster alphaproteobacteria (USCα)-like group. The copy numbers of pmoA genes ranged between 3.8 × 10⁵–1.9 × 10⁶ copies g⁻¹ of soil. Temperature was positively correlated with CH₄ uptake rates (P<0.001), but had no effect on methanotrophic population dynamics. The soil moisture was negatively correlated with CH₄ uptake rates (P<0.001), but showed a positive correlation with changes in USCα-like diversity (P<0.001) and pmoA gene abundance (P<0.05). These were greatest at low net CH₄ uptake rates during winter times and coincided with an overall increase in bacterial 16S rRNA gene abundances (P<0.05). Taken together, soil moisture had a significant but opposed effect on CH₄ uptake rates and methanotrophic population dynamics, the latter being increasingly stimulated by soil moisture contents >50 vol% and primarily related to members of the MHP clade.     

Diversity of methanotrophs in urea-fertilized tropical rice agroecosystem

Laboratory experiments were conducted to study the population size, diversity and methane oxidation potential of methanotrophs in tropical rice agroecosystem under the influence of N-fertilizer. Results indicate that the diversity of methane oxidizing bacteria (MOB) is altered in fertilizer treated soils compared to untreated control. Nevertheless, Type I MOB still dominated in the fertilized soils whereas the diversity of Type II methanotrophs decreases. Control soils have higher MOB population and CH₄ oxidation capacity than fertilized soils. Rhizospheric soil is more populated than non-rhizospheric soil in both unfertilized and fertilized conditions. Variation in K_m and V_max of methane oxidation in soils appears to be due to variation in methanotrophic community. Experimental results indicate that methanotrophic community differs both quantitatively and qualitatively in unfertilized and fertilized soils.     

Methanotrophic Activity,Abundance, and Diversity in Forested Swamp Pools: Spatiotemporal Dynamics and Influences on Methane Fluxes

Methane oxidation (methanotrophy) in the water column and sediments of forested swamp pools likely control seasonal and annual emission of CH₄ from these systems, but the methanotrophic microbial communities, their activities, locations, and overall impact, is poorly understood. Several techniques including ¹⁴CH₄ oxidation assays, culture-based most probable number (MPN) estimates of methane-oxidizing bacteria (MOB) and protozoan abundance, MOB strain isolation and characterization, and PCR techniques were used to investigate methanotrophy at a forested swamp near Ithaca, New York. The greatest methanotrophic activity and largest numbers of MOB occurred predominantly at the low oxygen sediment/water interface in the water column. Seasonally, methanotrophic activity was very dynamic, ranging from 0.1 to 61.9 μ moles CH₄ d^?1 g^?1 dry sediment, and correlated most strongly with dissolved inorganic carbon (r = 0.896). Incorporation of methanotrophic variables (abundance and activity) into existing CH₄ flux regression models improved model fit, particularly during mid summer when CH₄ fluxes were most dynamic. Annually integrated methane flux and methanotrophic activity measurements indicate that differences in methanotrophic activity at the sediment/water interface likely accounted for differences in the annual CH₄ emission from the field site. Direct isolations of MOB resulted in the repeated isolation of organisms most closely related to Methylomonas methanica S1. A single acidophilic, type II MOB related to Methylocella palustris K was also isolated. Using a PCR-based MPN method and 16S rRNA genome copy number from isolates and control strains, type I and type II MOB were enumerated and revealed type I dominance of the sediment-associated MOB community.     

陆地生态系统甲烷产生和氧化过程的微生物机理

陆地生态系统存在许多常年性或季节性缺氧环境,如:湿地、水稻土、湖泊沉积物、动物瘤胃、垃圾填埋场和厌氧生物反应器等。每年有大量有机物质进入这些环境,在缺氧条件下发生厌氧分解。甲烷是有机质厌氧分解的最终产物。产生的甲烷气体可通过缺氧-有氧界面释放到大气,产生温室效应,是重要的温室气体。产甲烷过程是缺氧环境中有机质分解的核心环节,而甲烷氧化是缺氧-有氧界面的重要微生物过程。甲烷的产生和氧化过程共同调控大气甲烷浓度,是全球碳循环不可分割的组成部分。对陆地生态系统甲烷产生和氧化过程的微生物机理研究进展进行了概要回顾和综述。主要内容包括:新型产甲烷古菌即第六和第七目产甲烷古菌和嗜冷嗜酸产甲烷古菌的发现;短链脂肪酸中间产物互营氧化过程与直接种间电子传递机制;新型甲烷氧化菌包括厌氧甲烷氧化菌和疣微菌属好氧甲烷氧化菌的发现;甲烷氧化菌生理生态与环境适应的新机制。这些研究进展显著拓展了人们对陆地生态系统甲烷产生和氧化机理的认识和理解。随着新一代土壤微生物研究技术的发展与应用,甲烷产生和氧化微生物研究领域将面临更多机遇和挑战,对未来发展趋势做了展望。     

The biogeochemical controls of N2O production and emission in landfill cover soils: the role of methanotrophs in the nitrogen cycle

Emissions of N2O from cover soils of both abandoned (> 30 years) and active landfills greatly exceed the maximum fluxes previously reported for tropical soils, suggesting high microbial activities for N2O production. Low soil matrix potentials (<-0.7 MPa) indicate that nitrification was the most likely mechanism of N2O formation during most of the time of sampling. Soil moisture had a strong influence on N2O emissions. The production of N2O was stimulated by as much as 20 times during laboratory incubations, when moisture was increased from -2.0 MPa to -0.6 MPa. Additional evidence from incubation experiments and delta13C analyses of fatty acids (18:1) diagnostic of methanotrophs suggests that N2O is formed in these soils by nitrification via methanotrophic bacteria. In a NH3(g)-amended landfill soil, the rate of N2O production was significantly increased when incubated with 100 ppmv methane compared with 1.8 ppmv (atmospheric) methane. Preincubation of a landfill soil with 1% CH4 for 2 weeks resulted in higher rates of N2O production when subsequently amended with NH3(g) relative to a control soil preincubated without CH4. At one location, at the soil depth (9-16 cm) of maximum methane consumption and N2O production, we observe elevated concentrations of organic carbon and nitrogen and distinct minima in delta15N (+1.0%) and delta13C (-33.8%) values for organic nitrogen and organic carbon respectively. A delta13C value of -39.3% was measured for 18:1 carbon fatty acids in this soil, diagnostic of type II methanotrophs. The low delta15N value for organic nitrogen is consistent with N2 fixation by type II methanotrophs. These observations all point to a methanotrophic origin for the organic matter at this depth. The results of this study corroborate previous reports of methanotrophic nitrification and N2O formation in aqueous and soil environments and suggest a predominance of type II rather than type I or type X methanotrophs in this landfill soil.     

Activity of a Methanotrophic Consortium Isolated from Landfill Cover Soil: Response to Temperature,pH, CO2, and Porous Adsorbent

A robust, naturally evolving methanotrophic community in landfill cover soil (LFCS) can be the simplest way to mitigate landfill methane emission. In this study, bacterial community composition in LFCS and methane oxidation potential of enriched methanotrophic consortium, in comparison to that of axenic Methylosinus sporium, was investigated. Growth and methane oxidation of the consortium was studied in liquid phase batch experiments under varying temperature (20–40°C), pH (5–10), headspace CO2, and in presence of porous adsorbent (1.3 cm³ sponge cubes). The 16S rRNA gene analysis revealed presence of both type-I and type-II methanotrophs along with few obligate methylotroph in LFCS. Though the optimal growth condition of the consortium was at 30°C and pH 7, it was more resilient in comparison to M. sporium. With increasing availability of porous adsorbent, methane consumption by the consortium was significantly improved (p < 0.001) reaching a maximum specific methane oxidation rate of 11.4 μmol mg^?1 biomass h^?1. Thus, inducing naturally thriving methanotrophs in LFCS is a better alternative to axenic methanotrophic culture in methane emission management.     

Increasing soil methane sink along a 120‐year afforestation chronosequence is driven by soil moisture

Upland soils are important sinks for atmospheric methane (CH₄), a process essentially driven by methanotrophic bacteria. Soil CH₄ uptake often depends on land use, with afforestation generally increasing the soil CH₄ sink. However, the mechanisms driving these changes are not well understood to date. We measured soil CH₄ and N₂O fluxes along an afforestation chronosequence with Norway spruce (Picea abies L.) established on an extensively grazed subalpine pasture. Our experimental design included forest stands with ages ranging from 25 to >120 years and included a factorial cattle urine addition treatment to test for the sensitivity of soil CH₄ uptake to N application. Mean CH₄ uptake significantly increased with stand age on all sampling dates. In contrast, CH₄ oxidation by sieved soils incubated in the laboratory did not show a similar age dependency. Soil CH₄ uptake was unrelated to soil N status (but cattle urine additions stimulated N₂O emission). Our data indicated that soil CH₄ uptake in older forest stands was driven by reduced soil water content, which resulted in a facilitated diffusion of atmospheric CH₄ into soils. The lower soil moisture likely resulted from increased interception and/or evapotranspiration in the older forest stands. This mechanism contrasts alternative explanations focusing on nitrogen dynamics or the composition of methanotrophic communities, although these factors also might be at play. Our findings further imply that the current dramatic increase in forested area increases CH₄ uptake in alpine regions.     

Diversity and activity of methanotrophs in landfill cover soils with and without landfill gas recovery systems

Aerobic CH₄ oxidation plays an important role in mitigating CH₄ release from landfills to the atmosphere. Therefore, in this study, oxidation activity and community of methanotrophs were investigated in a subtropical landfill. Among the three sites investigated, the highest CH₄ concentration was detected in the landfill cover soil of the site (A) without a landfill gas (LFG) recovery system, although the refuse in the site had been deposited for a longer time (∼14–15 years) compared to the other two sites (∼6–11 years) where a LFG recovery system was applied. In April and September, the higher CH₄ flux was detected in site A with 72.4 and 51.7 g m⁻² d⁻¹, respectively, compared to the other sites. The abundance of methanotrophs assessed by quantification of pmoA varied with location and season. A linear relationship was observed between the abundance of methanotrophs and CH₄ concentrations in the landfill cover soils (R = 0.827, P < 0.001). The key factors influencing the methanotrophic diversity in the landfill cover soils were pH, the water content and the CH₄ concentration in the soil, of which pH was the most important factor. Type I methanotrophs, including Methylococcus, Methylosarcina, Methylomicrobium and Methylobacter, and type II methanotrophs (Methylocystis) were all detected in the landfill cover soils, with Methylocystis and Methylosarcina being the dominant genera. Methylocystis was abundant in the slightly acidic landfill cover soil, especially in September, and represented more than 89% of the total terminal-restriction fragment abundance. These findings indicated that the LFG recovery system, as well as physical and chemical parameters, affected the diversity and activity of methanotrophs in landfill cover soils.     

Changing land use reduces soil CH4 uptake by altering biomass and activity but not composition of high-affinity methanotrophs

Forest ecosystems assimilate more CO₂ from the atmosphere and store more carbon in woody biomass than most nonforest ecosystems, indicating strong potential for afforestation to serve as a carbon management tool. However, converting grasslands to forests could affect ecosystem–atmosphere exchanges of other greenhouse gases, such as nitrous oxide and methane (CH₄), effects that are rarely considered. Here, we show that afforestation on a well-aerated grassland in Siberia reduces soil CH₄ uptake by a factor of 3 after 35 years of tree growth. The decline in CH₄ oxidation was observed both in the field and in laboratory incubation studies under controlled environmental conditions, suggesting that not only physical but also biological factors are responsible for the observed effect. Using incubation experiments with ¹³CH₄ and tracking ¹³C incorporation into bacterial phospholipid fatty acid (PLFA), we found that, at low CH₄ concentrations, most of the ¹³C was incorporated into only two PLFAs, 18 : 1ω7 and 16 : 0. High CH₄ concentration increased total ¹³C incorporation and the number of PLFA peaks that became labeled, suggesting that the microbial assemblage oxidizing CH₄ shifts with ambient CH₄ concentration. Forests and grasslands exhibited similar labeling profiles for the high-affinity methanotrophs, suggesting that largely the same general groups of methanotrophs were active in both ecosystems. Both PLFA concentration and labeling patterns indicate a threefold decline in the biomass of active methanotrophs due to afforestation, but little change in the methanotroph community. Because the grassland consumed CH₄ at a rate five times higher than forest soils under laboratory conditions, we concluded that not only biomass but also cell-specific activity was higher in grassland than in afforested plots. While the decline in biomass of active methanotrophs can be explained by site preparation (plowing), inorganic N (especially NH₄⁺) could be responsible for the change in cell-specific activity. Overall, the negative effect of afforestation of upland grassland on soil CH₄ uptake can be largely explained by the reduction in biomass and to a lesser extent by reduced cell-specific activity of CH₄-oxidizing bacteria.     

Interactive effects of drought and N fertilization on the spatial distribution of methane assimilation in grassland soils

Soil methanotrophic bacteria constitute the only globally relevant biological sink for atmospheric methane (CH₄). Nitrogen (N) fertilizers as well as soil moisture regime affect the activity of these organisms, but the mechanisms involved are not well understood to date. In particular, virtually nothing is known about the spatial distribution of soil methanotrophs within soil structure and how this regulates CH₄ fluxes at the ecosystem scale. We studied the spatial distribution of CH₄ assimilation and its response to a factorial drought × N fertilizer treatment in a 3‐year experiment replicated in two grasslands differing in management intensity. Intact soil cores were labelled with ¹⁴CH₄ and methanotrophic activity mapped at a resolution of ～100 μm using an autoradiographic technique. Under drought, the main zone of CH₄ assimilation shifted down the soil profile. Ammonium nitrate (NH₄NO₃) and cattle urine reduced CH₄ assimilation in the top soil, but only when applied under drought, presumably because NH₄⁺ from fertilizers was not removed by plant uptake and nitrification under these conditions. Ecosystem‐level CH₄ fluxes measured in the field did show no or only very small inhibitory effects, suggesting that deeper soil layers fully compensated for the reduction in top soil CH₄ assimilation. Our results indicate that the ecosystem‐level CH₄ sink cannot be inferred from measurements of soil samples that do not reflect the spatial organization of soils (e.g. stratification of organisms, processes, and mechanisms). The autoradiographic technique we have developed is suited to study methanotrophic activity in a relevant spatial context and does not rely on the genetic identity of the soil bacterial communities involved, thus ideally complementing DNA‐based approaches.