Ecosystem responses to increased precipitation and permafrost decay in subarctic Sweden inferred from peat and lake sediments

Recent accelerated decay of discontinuous permafrost at the Stordalen Mire in northern Sweden has been attributed to increased temperature and snow depth, and has caused expansion of wet minerotrophic areas leading to significant changes in carbon cycling in the mire. In order to track these changes through time and evaluate potential forcing mechanisms, this paper analyses a peat succession and a lake sediment sequence from within the mire, providing a record for the last 100 years, and compares these with monitored climate and active layer thickness data. The peat core was analysed for testate amoebae to reconstruct changes in peatland surface moisture conditions and water table fluctuations. The lake sediment core was analysed by near infrared spectroscopy to infer changes in the total organic carbon (TOC) concentration of the lake‐water, and changes in δ¹³C and C, N and δ¹⁵N to track changes in the dissolved inorganic carbon (DIC) pool and the influence of diagenetic effects on sediment organic matter, respectively. Results showed that major shifts towards increased peat surface moisture and TOC concentration of the lake‐water occurred around 1980, one to two decades earlier than a temperature driven increase in active layer thickness. Comparison with monitored temperature and precipitation from a nearby climate station indicates that this change in peat surface moisture is related to June–September (JJAS) precipitation and that the increase in lake‐water TOC concentration reflects an increase in total annual precipitation. A significant depletion in ¹³C of sediment organic matter in the early 1980s probably reflects the effect of a single or a few consecutive years with anomalously high summer precipitation, resulting in elevated DIC content of the lake water, predominantly originating from increased export and subsequent respiration of organic carbon from the mire. Based on these results, it was not possible to link proxy data obtained on peat and lake‐sediment records directly to permafrost decay. Instead our data indicate that increased precipitation and anomalously high rainfall during summers had a significant impact on the mire and the adjacent lake ecosystem. We therefore propose that effects of increased precipitation should be considered when evaluating potential forcing mechanisms of recent changes in carbon cycling in the subarctic.     

Methanogenesis from acetate: Physiology of a thermophilic, acetate-utilizing methanogenic bacterium

Abstract The biosynthesis of K88, K99 and F41 fibrillae by enterotoxigenic Escherichia coli strains was shown to be dependent on the growth phase of the cultures. An increase in adhesin production was observed, during exponential growth reaching its maximum at the end of this phase; thereafter adhesin production was arrested.
A simple and rapid purification procedure was developed for adhesins isolated from exponentially growing cells.     

Biogeochemistry of methane and methanogenic archaea in permafrost

This study summarizes the findings of our research on the genesis of methane, its content and distribution in permafrost horizons of different age and origin. Supported by reliable data from a broad geographical sweep, these findings confirm the presence of methane in permanently frozen fine-grained sediments. In contrast to the omnipresence of carbon dioxide in permafrost, methane-containing horizons (up to 40.0 mL kg(-1)) alternate with strata free of methane. Discrete methane-containing horizons representing over tens of thousands of years are indicative of the absence of methane diffusion through the frozen layers. Along with the isotopic composition of CH(4) carbon (delta(13)C -64 per thousand to -99 per thousand), this confirms its biological origin and points to in situ formation of this biogenic gas. Using (14)C-labeled substrates, the possibility of methane formation within permafrost was experimentally shown, as confirmed by delta(13)C values. Extremely low values (near -99 per thousand) indicate that the process of CH(4) formation is accompanied by the substantial fractionation of carbon isotopes. For the first time, cultures of methane-forming archaea, Methanosarcina mazei strain JL01 VKM B-2370, Methanobacterium sp. strain M2 VKM B-2371 and Methanobacterium sp. strain MK4 VKM B-2440 from permafrost, were isolated and described.     

Stress response of methanogenic archaea from Siberian permafrost compared with methanogens from nonpermafrost habitats

We examined the survival potential of methanogenic archaea exposed to different environmental stress conditions such as low temperature (down to −78.5°C), high salinity (up to 6 M NaCl), starvation (up to 3 months), long-term freezing (up to 2 years), desiccation (up to 25 days) and oxygen exposure (up to 72 h). The experiments were conducted with methanogenic archaea from Siberian permafrost and were complemented by experiments on well-studied methanogens from nonpermafrost habitats. Our results indicate a high survival potential of a methanogenic archaeon from Siberian permafrost when exposed to the extreme conditions tested. In contrast, these stress conditions were lethal for methanogenic archaea isolated from nonpermafrost habitats. A better adaptation to stress was observed at a low temperature (4°C) compared with a higher one (28°C). Given the unique metabolism of methanogenic archaea in general and the long-term survival and high tolerance to extreme conditions of the methanogens investigated in this study, methanogenic archaea from permafrost should be considered as primary candidates for possible subsurface Martian life.     

Interconnection of methanogenic and acetogenic pathways

Abstract Methanopterin displays a folate-like biochemistry: methenyl-, methylene- and methyltetrahydromethanopterin are intermediates in the process of methanogenesis. A corrinoid-enzyme is involved in methanol conversion and the possible function of such enzymes in cell-carbon synthesis and acetate conversion is discussed. Energy conservation is proposed to proceed via the establishment of a proton motive force through a vectorial reduction in the final step of methanogenesis and via substrate level phosphorylation in the oxidation of formyltetrahydromethanopterin. The biochemical reactions of methanogenesis and acetogenesis are compared. Methanogenic bacteria appear to use an aberrant set of coenzymes in almost all of the reactions involved in methanogenesis.     

Enzymology of one-carbon metabolism in methanogenic pathways

Methanoarchaea, the largest and most phylogenetically diverse group in the Archaea domain, have evolved energy-yielding pathways marked by one-carbon biochemistry featuring novel cofactors and enzymes. All of the pathways have in common the two-electron reduction of methyl-coenzyme M to methane catalyzed by methyl-coenzyme M reductase but deviate in the source of the methyl group transferred to coenzyme M. Most of the methane produced in nature derives from acetate in a pathway where the activated substrate is cleaved by CO dehydrogenase/acetyl-CoA synthase and the methyl group is transferred to coenzyme M via methyltetrahydromethanopterin or methyltetrahydrosarcinapterin. Electrons for reductive demethylation of the methyl-coenzyme M originate from oxidation of the carbonyl group of acetate to carbon dioxide by the synthase. In the other major pathway, formate or H2 is oxidized to provide electrons for reduction of carbon dioxide to the methyl level and reduction of methyl-coenzyme to methane. Methane is also produced from the methyl groups of methanol and methylamines. In these pathways specialized methyltransferases transfer the methyl groups to coenzyme M. Electrons for reduction of the methyl-coenzyme M are supplied by oxidation of the methyl groups to carbon dioxide by a reversal of the carbon dioxide reduction pathway. Recent progress on the enzymology of one-carbon reactions in these pathways has raised the level of understanding with regard to the physiology and molecular biology of methanogenesis. These advances have also provided a foundation for future studies on the structure/function of these novel enzymes and exploitation of the recently completed sequences for the genomes from the methanoarchaea Methanobacterium thermoautotrophicum and Methanococcus jannaschii.     

Plant-mediated effects of elevated ultraviolet-B radiation on peat microbial communities of a subarctic mire

Elevated ultraviolet‐B (UVB) radiation has been reported to have few effects on plants but to alter the soil microbial community composition. However, the effects on soil microorganisms have to be mediated via plants, because direct radiation effects are only plausible on the uppermost millimeters of soil. Here, we assessed secondary effects of UVB on soil microbes. The responses in the dominant plant Eriophorum russeolum, peat pore water and microbial communities in the peat were recorded at a subarctic mire in the middle of the third growing season under field exposure simulating 20% depletion in the ozone layer. The UVB treatment significantly reduced the sucrose and the total soluble sugar (sucrose+glucose+fructose) concentration of the plant leaves while increasing the sucrose concentration in the belowground storage organ rhizome. The starch concentration of the leaves was also slightly reduced by elevated UVB. In the plant roots, carbohydrate concentrations remained unaffected but the total phenolics concentration increased under elevated UVB. We suggest that the simultaneously observed decrease in bacterial growth rate and the altered bacterial community composition are due to UVB‐induced changes in the plant photosynthate allocation and potential changes in root exudation. There were no effects of elevated UVB on microbial biomass, peat pore water or nutrient concentrations in the peat. The observed responses are in line with the previously reported lower ecosystem dark respiration under elevated UVB, and they signify that the changed plant tissue quality and lower bacterial activity are likely to reduce decomposition.     

Acetoclastic and hydrogenotrophic methane production and methanogenic populations in an acidic West-Siberian peat bog

Sites in the West Siberian peat bog 'Bakchar' were acidic (pH 4.2-4.8), low in nutrients, and emitted CH4 at rates of 0.2-1.5 mmol m(-2) h(-1). The vertical profile of delta13CH4 and delta13CO2 dissolved in the porewater indicated increasing isotope fractionation and thus increasing contribution of H2/CO2-dependent methanogenesis with depth. The anaerobic microbial community at 30-50 cm below the water table produced CH4 with optimum activity at 20-25 degrees C and pH 5.0-5.5 respectively. Inhibition of methanogenesis with 2-bromo-ethane sulphonate showed that acetate, phenyl acetate, phenyl propionate and caproate were important intermediates in the degradation pathway of organic matter to CH4. Further degradation of these intermediates indicated that 62-72% of the CH4 was ultimately derived from acetate, the remainder from H2/CO2. Turnover times of [2-14C]acetate were on the order of 2 days (15, 25 degrees C) and accounted for 60-65% of total CH4 production. Conversion of 14CO2 to 14CH4 accounted for 35-43% of total CH4 production. These results showed that acetoclastic and hydrogenotrophic methanogenesis operated closely at a ratio of approximately 2 : 1 irrespective of the incubation temperature (4, 15 and 25 degrees C). The composition of the archaeal community was determined in the peat samples by terminal restriction fragment length polymorphism (T-RFLP) analysis and sequencing of amplified SSU rRNA gene fragments, and showed that members of Methanomicrobiaceae, Methanosarcinaceae and Rice cluster II (RC-II) were present. Other, presumably non-methanogenic archaeal clusters (group III, RC-IV, RC-V, RC-VI) were also detected. Fluorescent in situ hybridization (FISH) showed that the number of Bacteria decreased (from 24 x 10(7) to 4 x 10(7) cells per gram peat) with depth (from 5 to 55 cm below the water table), whereas the numbers of Archaea slightly increased (from 1 x 10(7) to 2 x 10(7) cells per gram peat). Methanosarcina spp. accounted for about half of the archaeal cells. Our results show that both hydrogenotrophic and acetoclastic methanogenesis are an integral part of the CH4-producing pathway in acidic peat and were represented by appropriate methanogenic populations.     

低温湿地甲烷古菌及其介导的甲烷产生途径

甲烷是重要的温室气体,低温湿地是大气甲烷的重要来源,因为湿地土壤中生活着大量的微生物包括甲烷古菌,它们将有机物降解转化为甲烷.本文总结了近年来低温湿地甲烷古菌群落组成、甲烷产生途径及其与环境的关系.研究显示,乙酸是低温湿地中主要的产甲烷物质,氢产甲烷过程主要发生在中温地区或酸性泥炭土中,而在盐碱水域中甲醇、甲胺是甲烷的重要底物.位于我国青藏高原的若尔盖湿地具有高海拔但低纬度的地理特征,我们的前期研究却显示甲醇在该湿地的甲烷排放中具有重要贡献.相应地,低温湿地中的甲烷古菌主要是利用甲基类化合物/乙酸的甲烷八叠球菌目和氢营养型的甲烷微球菌目.然而不同类型湿地甲烷排放途径及甲烷古菌的差异主要与环境的土壤类型、pH及植被类型相关,如刚毛荸荠生长的若尔盖湿地土壤中来源于甲醇的甲烷占全部甲烷的l7％;而木里苔草土壤中乙酸是产甲烷的主要前体物质.尽管已知冷适应的甲烷古菌在低温湿地的甲烷排放中发挥重要作用,但目前获得培养的嗜冷甲烷古菌却很少.冷响应的组学研究显示甲烷古菌的冷适应涉及到全局性生物学过程.     

Methanogenesis from Methylated Amines in a Hypersaline Algal Mat

Methane ebullition and high rates of methane production were observed in sediments of a hypersaline pond (180‰) which contained sulfate in excess of 100 mM. The highest rates of methane production were observed in surface sediments associated with an algal mat dominated by a Spirulina sp. The mat contained a methylated amine, glycine betaine (GBT), at levels which accounted for up to 20% of the total mat nitrogen. GBT was apparently the source of trimethylamine (TMA), which was also present in the sediment at relatively high concentrations. Patterns of substrate metabolism by the methanogenic populations in sediment slurries suggested that TMA was a major methane precursor. Neither exogenous hydrogen nor acetate stimulated methanogenesis, while addition of a variety of amines including TMA, trimethylamine oxide, GBT, and choline resulted in substantial increases with yields of >70%. The temperature optimum for methanogenesis in this system was 45 to 55°C, which coincided with the observed sediment temperature. Patterns and rates of methane production in this and other hypersaline algal mats may be determined by a complex interaction between salinity, the use of methylated amines for osmoregulation by algae, and the formation of TMA by fermentation.     

Propionate metabolism in a methanogenic enrichment culture. Direct reductive carboxylation and acetogenesis pathways

Abstract Serial dilutions of methanogenic sludges in propionate medium gave a methanogenic non-acetoclastic enrichment degrading 1 mol of propionate to 1.6 mol of acetate and 0.17 mol of methane, with a transient accumulation of butyrate. NMR recordings showed the conversion of [2-¹³C]- and [3-¹³C]-propionate to [3-¹³C]- and [4-¹³C]-butyrate, respectively, thus demonstrating a reductive carboxylation of propionate to butyrate. The labelling found in the accumulated acetate and fermentation balances also suggested that reductive carboxylation was the major pathway involved in propionate conversion to acetate.     

Characterization of acid-tolerant H/CO-utilizing methanogenic enrichment cultures from an acidic peat bog in New York State

Two methanogenic cultures were enriched from acidic peat soil using a growth medium buffered to c. pH 5. One culture, 6A, was obtained from peat after incubation with H(2)/CO(2), whereas culture NTA was derived from a 10(-4) dilution of untreated peat into a modified medium. 16S rRNA gene clone libraries from each culture contained one methanogen and two bacterial sequences. The methanogen 16S rRNA gene sequences were 99% identical with each other and belonged to the novel "R-10/Fen cluster" family of the Methanomicrobiales, whereas their mcrA sequences were 96% identical. One bacterial 16S rRNA gene sequence from culture 6A belonged to the Bacteroidetes and showed 99% identity with sequences from methanogenic enrichments from German and Russian bogs. The other sequence belonged to the Firmicutes and was identical to a thick rod-shaped citrate-utilizing organism isolated from culture 6A, the numbers of which decreased when the Ti (III) chelator was switched from citrate to nitrilotriacetate. Bacterial clones from the NTA culture clustered in the Delta- and Betaproteobacteria. Both cultures contained thin rods, presumably the methanogens, as the predominant morphotype, and represent a significant advance in characterization of the novel acidiphilic R-10 family methanogens.     

Temperature and peat type control CO2 and CH4 production in Alaskan permafrost peats

Controls on the fate of ~277 Pg of soil organic carbon (C) stored in permafrost peatland soils remain poorly understood despite the potential for a significant positive feedback to climate change. Our objective was to quantify the temperature, moisture, organic matter, and microbial controls on soil organic carbon (SOC) losses following permafrost thaw in peat soils across Alaska. We compared the carbon dioxide (CO₂) and methane (CH₄) emissions from peat samples collected at active layer and permafrost depths when incubated aerobically and anaerobically at ?5, ?0.5, +4, and +20 °C. Temperature had a strong, positive effect on C emissions; global warming potential (GWP) was >3× larger at 20 °C than at 4 °C. Anaerobic conditions significantly reduced CO₂ emissions and GWP by 47% at 20 °C but did not have a significant effect at ?0.5 °C. Net anaerobic CH₄ production over 30 days was 7.1 ± 2.8 μg CH₄‐C gC^?1 at 20 °C. Cumulative CO₂ emissions were related to organic matter chemistry and best predicted by the relative abundance of polysaccharides and proteins (R² = 0.81) in SOC. Carbon emissions (CO₂‐C + CH₄‐C) from the active layer depth peat ranged from 77% larger to not significantly different than permafrost depths and varied depending on the peat type and peat decomposition stage rather than thermal state. Potential SOC losses with warming depend not only on the magnitude of temperature increase and hydrology but also organic matter quality, permafrost history, and vegetation dynamics, which will ultimately determine net radiative forcing due to permafrost thaw.     

A frozen feast: thawing permafrost increases plant‐available nitrogen in subarctic peatlands

Many of the world's northern peatlands are underlain by rapidly thawing permafrost. Because plant production in these peatlands is often nitrogen (N)‐limited, a release of N stored in permafrost may stimulate net primary production or change species composition if it is plant‐available. In this study, we aimed to quantify plant‐available N in thawing permafrost soils of subarctic peatlands. We compared plant‐available N‐pools and ‐fluxes in near‐surface permafrost (0–10 cm below the thawfront) to those taken from a current rooting zone layer (5–15 cm depth) across five representative peatlands in subarctic Sweden. A range of complementary methods was used: extractions of inorganic and organic N, inorganic and organic N‐release measurements at 0.5 and 11 °C (over 120 days, relevant to different thaw‐development scenarios) and a bioassay with Poa alpina test plants. All extraction methods, across all peatlands, consistently showed up to seven times more plant‐available N in near‐surface permafrost soil compared to the current rooting zone layer. These results were supported by the bioassay experiment, with an eightfold larger plant N‐uptake from permafrost soil than from other N‐sources such as current rooting zone soil or fresh litter substrates. Moreover, net mineralization rates were much higher in permafrost soils compared to soils from the current rooting zone layer (273 mg N m^?2 and 1348 mg N m^?2 per growing season for near‐surface permafrost at 0.5 °C and 11 °C respectively, compared to ?30 mg N m^?2 for current rooting zone soil at 11 °C). Hence, our results demonstrate that near‐surface permafrost soil of subarctic peatlands can release a biologically relevant amount of plant available nitrogen, both directly upon thawing as well as over the course of a growing season through continued microbial mineralization of organically bound N. Given the nitrogen‐limited nature of northern peatlands, this release may have impacts on both plant productivity and species composition.     

Methanogenesis and sulfate reduction in timber and drainage water from a gold mine

Biogenesis of methane in the heartwood of diseased trees has been shown, but never in timber in service. Studies were undertaken to establish whether methan‐ogens and sulfate‐reducers were present in wooden pit props and drainage water from underground sites in a gold mine. The predominant methanogen in the mine ecosystem was tentatively identified as Methanobacterium bryantii. The sulfate‐reducers comprised Desulfovibrio desulfuricans and Desulfotomaculum antarcticum. Most probable numbers (MPN) of bacteria indicated that 3.5 × 10⁵ methanogenic and 7.9 × 10³ sulfate‐reducing bacteria were present per milliliter of stagnant drainage water. MPN values per gram of timber were lower for methanogens but comparable for sulfate‐reducers. Laboratory model systems predicted a maximum rate of methan‐ogenesis of 2.3 mL methane/g wood per day; however, rates would never attain this value because of nutrient limitations and environmental restrictions. Analysis of gas samples extracted from sealed areas of the gold mine verified the presence of methane.     

Shifts of methanogenic communities in response to permafrost thaw results in rising methane emissions and soil property changes

Extremophiles - Permafrost thaw can bring negative consequences in terms of ecosystems, resulting in permafrost collapse, waterlogging, thermokarst lake development, and species composition...     

Methanogenesis from acetate: a nonmethanogenic bacterium from an anaerobic acetate enrichment

A methanogenic acetate enrichment was initiated by inoculation of an acetate-mineral salts medium with domestic anaerobic digestor sludge and maintained by weekly transfer for 2 years. The enrichment culture contained a Methanosarcina and several obligately anaerobic nonmethanogenic bacteria. These latter organisms formed varying degrees of association with the Methanosarcina, ranging from the nutritionally fastidious gram-negative rod called the satellite bacterium to the nutritionally nonfastidious Eubacterium limosum. The satellite bacterium had growth requirements for amino acids, a peptide, a purine base, vitamin B12, and other B vitamins. Glucose, mannitol, starch, pyruvate, cysteine, lysine, leucine, isoleucine, arginine, and asparagine stimulated growth and hydrogen production. Acetate was neither incorporated nor metabolized by the satellite organism. Since acetate was the sole organic carbon source in the enrichment culture, organism(s) which metabolize acetate (such as the Methanosarcina) must produce substrates and growth factors for associated organisms which do not metabolize acetate.     

Methanogenesis from lactate by a co-culture of Clostridium formicoaceticum and Methanosarcina mazei

Summary A co-culture of Clostridium formicoaceticum and Methanosarcina mazei converted lactate to methane and carbon dioxide at mesophilic temperatures and pH values near 7.0. Lactate was first converted to acetate by the homoacetogen, and then to CH₄ and CO₂ by the methanogen, with the second reaction as the rate-limiting step. The methane yield was about 1.45 mol/mol lactate. These two organisms formed a mutualistic association and may be useful together with the homolactic bacterium Stretococcus lactis to convert lactose to methane. Offprint requests to: S. T. Yang