Methane concentrations and methanotrophic community structure influence the response of soil methane oxidation to nitrogen content in a temperate forest

Methane oxidation in forest soils removes atmospheric CH₄. Many studies have determined methane uptake rates and their controlling variables, yet the microorganisms involved have rarely been assessed simultaneously over the longer term. We measured methane uptake rates and the community structure of methanotrophic bacteria in temperate forest soil (sandy clay loam) on a monthly basis for two years in South Korea. Methane uptake rates at the field site did not show any seasonal patterns, and net uptake occurred throughout both years. In situ uptake rates and uptake potentials determined in the laboratory were 2.92 ± 4.07 mg CH₄ m⁻² day⁻¹ and 51.6 ± 45.8 ng CH₄ g⁻¹ soil day⁻¹, respectively. Contrary to results from other studies, in situ oxidation rates were positively correlated with soil nitrate concentrations. Short-term experimental nitrate addition (0.20-1.95 μg N g⁻¹ soil) significantly stimulated oxidation rates under low methane concentrations (1.7-2.0 ppmv CH₄), but significantly inhibited oxidation under high methane concentrations (300 ppmv CH₄). We analyzed the community structures of methanotrophic bacteria using a DNA-based fingerprinting method (T-RFLP). Type II methanotrophs dominated under low methane concentrations while Type I methanotrophs dominated under high methane concentrations. Nitrogen addition selectively inhibited Type I methanotrophic bacteria. Overall, the results of this study indicate that the effects of inorganic N on methane uptake depend on methane concentrations and that such a response is related to the dissimilar activation or inhibition of different types of methanotrophic bacteria.     

Effect of moisture,texture and aggregate size of paddy soil on production and consumption of CH4

Laboratory experiments were conducted to find out under which conditions the soil from Italian rice fields could change from a source into a sink of atmospheric CH₄. Moist (30% H₂O=68% of the maximum water holding capacity (whc)) rice field soil oxidized CH₄ with biphasic kinetics, exhibiting both a low (145 ppmv CH₄) and a high (20,200 ppmv CH₄) K_m value and V_max values of 16.8 and 839 nmol gdw⁻¹ h⁻¹, respectively. The activity with the low K_m allowed the oxidation of atmospheric CH₄. Uptake rates of high CH₄ concentrations (16.5% v/v) and of O₂ linearly decreased with aggregate size of soil between 2 and 10 mm. Atmospheric CH₄ (1.8 ppmv) was consumed in soil aggregates <6 mm, but soil aggregates >6 mm released CH₄ into the atmosphere. Similarly, net uptake of atmospheric CH₄ turned into net release of CH₄ when the soil moisture was decreased below a water content of about 20% whc. The uptake rate of atmospheric CH₄ increased threefold when the soil was amended with sterile quartz sand. Flooded microcosms with non-amended and quartz-amended soil emitted CH₄ into the atmosphere. The CH₄ emission rate increased when the flux was measured under an atmosphere of N₂ instead of air, indicating that 30–99% of the produced CH₄ was oxidized in the oxic soil surface layer. Removal of the flood water resulted in increase of CH₄ emission rates until a water content of about 75–82% whc was reached, and subsequently in a rapid decrease. However, the soil microcosms never showed net uptake of atmospheric CH₄. Our results show that the microorganisms consuming atmospheric CH₄ were inactivated at an earlier stage of drainage than the microorganisms producing CH₄, irrespective of the soil porosity which was adjusted by addition of quartz sand. Hence, it is unlikely that the Italian rice fields can act as a net sink for atmospheric CH₄ even when drained.     

Methanotrophic communities in a landfill cover soil as revealed by [13C] PLFAs and respiratory quinones: Impact of high methane addition and landfill leachate irrigation

The soil microbial communities of a landfill cover substrate, which was treated with landfill gas (100 l CH₄ m^?2 d^?1) and landfill leachate for 1.5 years, were investigated by phospholipid fatty acid (PLFA), ergosterol and respiratory quinone analyses. The natural ¹³C depletion of methane was used to assess the activity of methanotrophs and carbon turnover in the soil system. Under methane addition, the soil microbial community was dominated by PLFAs (14:0 and 16:1 isomers) and quinones (ubiquinone-8 and 18-methylene-ubiquinone-8) related to type I methanotrophs, and 18:1 PLFAs contained in type II methanotrophs. While type I methanotrophic PLFAs were ¹³C depleted, i.e. type I methanotrophs were actively oxidising and assimilating methane, ¹³C depletion of 18:1 PLFAs was low and inconsistent with their abundance. This, possibly reflects isotopic discrimination, assimilation of carbon derived from type I methanotrophs and a high contribution of non-methanotrophic bacteria to the 18:1 isomers. Landfill leachate irrigation caused the methanotrophic community to shift closer to the soil surface. It also decreased 18:1 PLFAs, while type I methanotrophs were probably stimulated. Gram positive bacteria, but not fungi, were also ¹³C depleted and consequently involved in the secondary turnover of carbon originating from methanotrophic bacteria. Cy17:0 PLFA was ¹³C depleted in deep soil layers, indicating anaerobic methane oxidation.     

Soil methane oxidation and methanotroph responses to afforestation of pastures with Pinus radiata stands

Afforestation of pastures in New Zealand reduces methane (CH₄) production from soil, while also stimulating oxidation of atmospheric CH₄ by soil methanotrophs. However, neither the mechanisms by which soil CH₄ oxidation is enhanced by afforestation, nor how long after forest planting tree-dependent responses in CH₄ oxidation become detectable are fully known. Here, we investigated the effects of different-aged stands (5-20 y) of the exotic pine (Pinus radiata (D. Don)) on CH₄ oxidation and methanotrophic community structure in soils, compared with adjacent, long-established pastures. Two of the pastures were on volcanic soils and two were on non-volcanic soils. Although the CH₄ fluxes in soils from these young stands were not significantly different from those in the associated pastures, the rate of oxidation of added ¹³CH₄ was higher in the pine soils. Both fluxes and ¹³CH₄ oxidation rates were higher in the volcanic than the non-volcanic soils. Combined phospholipid fatty acid (PLFA) and stable isotope probe (SIP) analyses suggested that type II methanotrophs (PLFA C18:1ω7) were most active in all soils followed by uncultivable bacteria (C17:0ai). Molecular analysis of the methanotrophic community structure using pmoA (particulate methane monooxygenase) genes suggested that a particular type II methanotroph (TRF 35) was dominant in all soils, but more so in the pine than in pasture soils. A type I methanotroph (TRF 245) was more prevalent in the pasture than in associated pine soils, whereas TRF 128 (a type II methanotroph) was slightly more dominant in soils under pine. Cloning and sequencing data suggest TRFs 35 and 128, which differ from one another, belong to distant relatives of Methylocapsa sp; TRF 245 is related to Methylococcus capsulatus. Land-use change resulted in changes in soil bulk density, porosity, moisture contents and in methanotrophic community structure. Methane oxidation rates were most closely related to soil moisture, as well as to the methanotrophic community structure, and nitrate-N, extractable C and total C concentrations. Stepwise multiple regression also suggested a weak effect (P = 0.06) of stand age on CH₄ oxidation rate. By contrast, the responses of the methanotrophic community structure to this land-use change were more readily detected by the specific molecular analyses, and indicated a predominance of type II methanotrophs in pine soils.     

Soil–atmosphere greenhouse gas exchange in a cool,temperate Eucalyptus delegatensis forest in south-eastern Australia

Forests are the largest C sink (vegetation and soil) in the terrestrial biosphere and may additionally provide an important soil methane (CH₄) sink, whilst producing little nitrous oxide (N₂O) when nutrients are tightly cycled. In this study, we determine the magnitude and spatial variation of soil–atmosphere N₂O, CH₄ and CO₂ exchange in a Eucalyptus delegatensis forest in New South Wales, Australia, and investigate how the magnitude of the fluxes depends on the presence of N₂-fixing tree species (Acacia dealbata), the proximity of creeks, and changing environmental conditions. Soil trace gas exchange was measured along replicated transects and in forest plots with and without presence of A. dealbata using static manual chambers and an automated trace gas measurement system for 2 weeks next to an eddy covariance tower measuring net ecosystem CO₂ exchange. CH₄ was taken up by the forest soil (?51.8 μg CH₄-C m^?2 h^?1) and was significantly correlated with relative saturation (S_r) of the soil. The soil within creek lines was a net CH₄ source (up to 33.5 μg CH₄-C m^?2 h^?1), whereas the wider forest soil was a CH₄ sink regardless of distance from the creek line. Soil N₂O emissions were small (<3.3 μg N₂O-N m^?2 h^?1) throughout the 2-week period, despite major rain and snowfall. Soil N₂O emissions only correlated with soil and air temperature. The presence of A. dealbata in the understorey had no influence on the magnitude of CH₄ uptake, N₂O emission or soil N parameters. N₂O production increased with increasing soil moisture (up to 50% S_r) in laboratory incubations and gross nitrification was negative or negligible as measured through ¹⁵N isotope pool dilution.The small N₂O emissions are probably due to the limited capacity for nitrification in this late successional forest soil with C:N ratios >20. Soil–atmosphere exchange of CO₂ was several orders of magnitude greater (88.8 mg CO₂-C m^?2 h^?1) than CH₄ and N₂O, and represented 43% of total ecosystem respiration. The forest was a net greenhouse gas sink (126.22 kg CO₂-equivalents ha^?1 d^?1) during the 2-week measurement period, of which soil CH₄ uptake contributed only 0.3% and N₂O emissions offset only 0.3%.     

Soil–methane sink increases with soil age in forefields of Alpine glaciers

In upland soils aerobic methane-oxidizing bacteria (MOB) catalyze methane (CH₄) oxidation, and thus regulate the sole terrestrial sink for atmospheric CH₄. While confirmed in mature upland soils, little is known about this important function in young mountainous soils in glacier forefields, which are progressively formed as a result of glacier recession. We assessed four attributes of the soil CH₄ sink, i.e., soil-atmosphere CH₄ flux (J_atm), CH₄ oxidation activity (k), MOB abundance and variation in community composition along the 6–120-yr soil chronosequence in two Alpine glacier forefields on siliceous and calcareous bedrock. At most sampling locations soil CH₄ profiles showed stable uptake of atmospheric CH₄, with J_atm in the range of −0.082 to −2.2 mg CH₄ m⁻² d⁻¹. Multiple-linear-regression analyses indicated that J_atm significantly increased with soil age, whereas k did not. Instead, water content and CH₄ profiles in the youngest soils often indicated dry, inactive top layers with k < 0.1 h⁻¹, and active deeper layers (0.2 h⁻¹ ≤ k ≤ 11 h⁻¹) with more favorable water content. With increasing soil age the zone of highest CH₄ oxidation activity gradually moved upwards and eventually focused in the 10–40-cm layer (0.2 h⁻¹ ≤ k ≤ 16 h⁻¹). Copy numbers of pmoA genes significantly increased with soil age at both sites, ranging from 2.4 × 10³ to 5.5 × 10⁵ copies (g soil w.w.)⁻¹, but also correlated with mineral nitrogen content. Terminal restriction-fragment-length-polymorphism and cluster analyses showed differences in MOB community composition apparently related to bedrock type rather than soil age. Yet, regardless of bedrock type, the soil CH₄ sink established within a few years of soil development, and J_atm increased to values comparable to mature soils within decades. Thus, young mountainous soils have the potential to consume substantial amounts of atmospheric CH₄, and should be incorporated into future estimates of global soil CH₄ uptake.     

Tree species affect atmospheric CH4 oxidation without altering community composition of soil methanotrophs

Plant species exert strong effects on ecosystem functions and one of the emerging, and difficult to test hypotheses, is that plants alter soil functions through changing the community structure of soil microorganisms. We tested the hypothesis for atmospheric CH₄ oxidation by using soil samples from a Siberian afforestation experiment and exposing them to ¹³C-CH₄. We determined the activity of the soil methanotrophs under different tree species at three levels of initial CH₄ concentration (30, 200 and 1000 ppm) thus distinguishing the activities of low- and high-affinity methanotrophs. Half of the samples were incubated with ¹³C-enriched CH₄ (99.9%) and half with ¹²C-CH₄. This allowed an estimation of the amount of ¹³C incorporated into individual PLFAs and determination of PLFAs of methanotrophs involved in CH₄ oxidation at the different CH₄ concentrations. Tree species strongly altered the activity of atmospheric CH₄ oxidation without appearing to change the composition of high-affinity methanotrophs as evidenced by PLFA ¹³C labeling. The low diversity of atmospheric CH₄ oxidizers, presumably belonging to the UCSα group, may explain the lack of tree species effects on the composition of soil methanotrophs. We submit that the observed tree species effects on atmospheric CH₄ oxidation indicate an effect on biomass or cell-specific activities rather than by a community change and this may be related to the impact of the tree species on soil N cycling.     

The effects of acid nitrogen and acid sulphur deposition on CH4 oxidation in a forest soil: a laboratory study

Sieved soil and soil core experiments were performed to determine the potential sensitivity of forest soil CH₄ oxidation to oxidised N, reduced N and oxidised S atmospheric deposition. Ammonium sulphate was used to simulate reduced N deposition, HNO₃ oxidised N deposition and H₂SO₄ oxidised S deposition. The effects of NH₄⁺, NO₃⁻, SO₄²⁻ and H⁺ on soil CH₄ flux were shown to be governed by the associated counter-anion or cation of the investigated ions. Ammonium sulphate, at concentrations greater than those that would be experienced in polluted throughfall, showed a low potential to cause inhibition of CH₄ oxidation. In contrast, HNO₃ strongly inhibited net CH₄ oxidation in sieved soils and also in soil cores. In addition, soil CO₂ production was inhibited and the organic and mineral soil horizons acidified in HNO₃ treated soil cores. This suggested that the HNO₃ effect on CH₄ flux might be indirectly mediated through aluminium toxicity. Sulphuric acid only inhibited CH₄ oxidation when added at pH 1. At concentrations more representative of heavily polluted throughfall, H₂SO₄ had no effect on soil CH₄ flux or CO₂ production from soil cores, even after 210 days of repeated addition. In contrast to HNO₃ additions, acidification of the soil was not marked and was only significant for the mineral soil. The findings suggest that the response of forest soil CH₄ oxidation to atmospheric acid deposition is strongly dependent on the form of acid deposition.     

Conversion of cropland to forest increases soil CH4 oxidation and abundance of CH4 oxidizing bacteria with stand age

We investigated CH₄ oxidation in afforested soils over a 200-year chronosequence in Denmark including different tree species (Norway spruce, oak and larch) and ages. Samples of the top mineral soil (0–5 cm and 5–15 cm depth) were incubated and analyzed for the abundance of the soil methane-oxidizing bacteria (MOB) and ammonia-oxidizing bacteria (AOB) and archaea (AOA) based on quantitative PCR (qPCR) on pmoA and amoA genes. Our study showed that CH₄ oxidation rates and the abundance of MOB increased simultaneously with time since afforestation, suggesting that the methanotrophic activity is reflected in the abundance of this functional group.The development of forest soils resulted in increased soil organic carbon and reduced bulk density, and these were the two variables that most strongly related to CH₄ oxidation rates in the forest soils. For the top mineral soil layer (0–5 cm) CH₄ oxidation rates did not differ between even aged stands from oak and larch, and were significantly smaller under Norway spruce. Compared to the other tree species Norway spruce caused a decrease in the abundance of MOB over time that could explain the decreased oxidation rates. However, the cause for the lower abundance remains unclear. The abundance of ammonia-oxidizers along the chronosequence decreased over time, oppositely to the MOB. However, our study did not indicate a direct link between CH₄ oxidation rates and ammonia-oxidizers. Here, we provide evidence for a positive impact of afforestation of former cropland on CH₄ oxidation capacity in soils most likely caused by an increased population size and activity of MOB.     

Soil organic carbon and nitrogen in a Mollisol in central Ohio as affected by tillage and land use

Minimum tillage practices are known for increasing soil organic carbon (SOC). However, not all environmental situations may manifest this potential change. The SOC and N stocks were assessed on a Mollisol in central Ohio in an 8-year-old tillage experiment as well as under two relatively undisturbed land uses; a secondary forest and a pasture on the same soil type. Cropped systems had 51±4 (equiv. mass) Mg ha⁻¹ lower SOC and lower 3.5±0.3 (equiv. mass) Mg ha⁻¹ N in the top 30 cm soil layer than under forest. Being a secondary forest, the loss in SOC and N stocks by cultivation may have been even more than these reported herein. No differences among systems were detected below this depth. The SOC stock in the pasture treatment was 29±3 Mg ha⁻¹ greater in the top 10 cm layer than in cultivated soils, but was similar to those under forest and no-till (NT). Among tillage practices (plow, chisel and NT) only the 0–5 cm soil layer under NT exhibited higher SOC and N concentrations. An analysis of the literature of NT effect on SOC stocks, using meta-analysis, suggested that NT would have an overall positive effect on SOC sequestration rate but with a greater variability of what was previously reported. The average sequestration rate of NT was 330 kg SOC ha⁻¹ year⁻¹ with a 95% confidence interval ranging from 47 to 620 kg SOC ha⁻¹ year⁻¹. There was no effect of soil texture or crop rotation on the SOC sequestration rate that could explain this variability. The conversion factor for SOC stock changes from plow to NT was equal to 1.04. This suggests that the complex mechanisms and pathways of SOC accrual warrant a cautious approach when generalizing the beneficial changes of NT on SOC stocks.     

Emissions of CH4, CO2, and N2O from soil at a cattle overwintering area as affected by available C and N

Relationships between CH₄, CO₂, and N₂O emissions were studied in soil that had been freshly amended with large deposits of cattle wastes. Dynamics of CH₄, CO₂, and N₂O emissions were investigated with flux chambers from early April to late June 2011, during the 3 months following cattle overwintering at the site. This 81-day field study was supplemented with soil analyses of available C and N content and measurement of denitrification activity. In a more detailed field investigation, the daily time course of emissions was determined. The field research was complemented with a laboratory experiment that focused on the short-term time course of N₂O and CH₄ production in artificially created anoxic soil microsites. The following hypotheses were tested: (i) a large input of C (and N and other nutrients) in cattle manure creates conditions suitable for methanogenesis, and therefore overwintering areas can produce large amounts of CH₄; (ii) N₂O is produced and emitted until the level of mineral N decreases, while the level of CH₄ production is low; and (iii) production of CH₄ is greater when N immobilization decreases the level of NO₃⁻ in soil. N₂O emissions were relatively large during the first 3 weeks, then peaked (at ca. 4000 μg N₂ON m⁻² h⁻¹) and soon decreased to almost zero; the changes were related to the mineral and soluble organic N content in soil. CH₄ fluxes were large, though variable, in the first 2 months (600–3000 μg CH₄C m⁻² h⁻¹) and were independent of C and N availability. Although time courses differed for CH₄ and N₂O, a negative relationship between N₂O and CH₄ emissions was not detected. Contrary to CH₄ and N₂O fluxes, CO₂ emissions progressively increased to ca. 300 mg CO₂C m⁻² h⁻¹ at the end of the field study and were closely related to air and soil temperatures. Diurnal measurements revealed significant correlations between temperature and emissions of CH₄, N₂O, and CO₂. Addition of C to soil during anaerobic incubation increased the production and consumption of N₂O and supported the emission of CH₄. The results suggest that rapid denitrification significantly contributes to the exhaustion of oxidizing agents and helps create microsites supporting methanogenesis in otherwise N₂O-producing upland soil. The results also indicate that accurate estimate of gas fluxes in animal-impacted grassland areas requires assessment of both diurnal and long-term changes in CH₄, CO₂, and N₂O emissions.     

Ethylene oxidation,atmospheric methane consumption,and ammonium oxidation in temperate volcanic forest soils

Temperate volcanic forest surface soils under different forest stands (e.g., Pinus sylvestris L., Cryptomeria japonica, and Quercus serrata) were sampled to study the kinetics of ethylene (C₂H₄) oxidation and the C₂H₄ concentrations that effectively inhibit oxidation of atmospheric methane (CH₄) and nitrification. The kinetics of C₂H₄ oxidation in temperate volcanic forest soils was biphasic, indicating that at least two different microbial populations, one with low and another with high apparent K _m values, were responsible for ethylene oxidation. Methane consumption activity and ammonium oxidation of soil were inhibited by adding ethylene. Added C₂H₄ at concentrations of 3, 10, and 20 μl C₂H₄ per liter in the headspace gas respectively reduced by 20%, 50%, and 100% atmospheric CH₄ consumption by soil, and these values were much smaller than those inhibiting ammonium oxidation in these forest soils; thus, the CH₄ consumption activity was more sensitive to the addition of C₂H₄ than ammonium oxidation. Previous studies have shown that accumulation of C₂H₄ in such volcanic forest soils within 3 days of aerobic and anaerobic incubations can reach a range from 0.2 to 0.3 and from 1.0 to 3.0 μl C₂H₄ per liter in the headspace gas, respectively. It is suggested that C₂H₄ production beneath forest floors, particularly after heavy rain, can to some extent affect the capacity of forest surface soils to consume atmospheric CH₄, but probably, it has no impact on ammonium oxidation.     

Effect of wetting intensity on soil GHG fluxes and microbial biomass under a temperate forest floor during dry season

The increasing frequency of periodic droughts followed by heavy rainfalls is expected for this current century, but little is known about the effects of wetting intensity on the in situ biogenic greenhouse gas (GHG) fluxes of forest soils and soil microbial biomass. To gain new insights into the underlying mechanisms responsible for wetting-induced GHG fluxes in situ, rain simulation field experiments during a natural prolonged drought period were done under a temperate forest in northeast China. The intensity of rainfall-induced CO₂ pulses increased from 0.84 to 2.08 g CO₂–C m^? 2 d^? 1 with the intensity of wetting up to ca. 80% water-filled pore space, which coincided with an increase in soil microbial biomass and with a decrease in soil labile organic C following wetting. Methane uptake rates decreased from 1.76 to 0.87 mg CH₄–C m^? 2 d^? 1 with the intensity of wetting. Wetting dry forest floor increased N₂O fluxes from 6.2 to 25.9 μg N₂O–N m^? 2 d^? 1, but there was no significant difference between all experimental wetted plots. The rainfall-induced N₂O pulses with increasing wetting intensity were opposite to that of the CO₂ pulses, showing a maximum response at the lowest wetting intensity. An analysis of the temperature sensitivity of GHG fluxes indicated that temperature had an increased effect on the in situ CO₂ flux and CH₄ uptake, respectively, under wetted and dry conditions. The global warming potential of GHG fluxes and Q₁₀ value of the temperature response of CO₂ fluxes increased linearly with wetting intensity. The results indicate that the rainfall-induced soil CO₂ pulse is mainly due to enhanced microbial consumption on substrates and highlight the complex nature of belowground C-cycling responses to climate change in northeast China forests that normally experience periodic droughts followed by heavy rainfalls over the year.     

Temporal variation of soil compaction and deterioration of soil quality in pasture areas of Colombian Amazonia

In the Amazon basin, tropical rainforest is being slashed and burned at accelerated rates for annual crops over a couple of years, followed by forage grasses. Because of poor management, the productivity of established pastures declines in a few years so that grazing plots are abandoned and new areas are deforested. Previous studies in the region report higher bulk density in soils under pasture than in similar soils under forest. The objective of this study was to detect changes in the physical quality of the topsoil of nutrient-poor Typic Paleudults in the colonisation area of Guaviare, Colombian Amazonia, and analyse the effect of soil deterioration on pasture performance. Temporal variation of soil compaction under pasture was analysed by comparing natural forest taken as control and pasture plots of Brachiaria decumbens (Stapf) grouped into three age ranges (<3, 3–9, >9 years). Evidence of soil compaction through cattle trampling, after clearing the primary forest, included the formation of an Ap horizon with platy structure and dominant greyish or olive colours, reflecting impaired surface drainage, the increase of bulk density and penetration resistance, and the decrease of porosity and infiltration rate. From primary forest to pastures older than 9 years, bulk density of the 5–10 cm layer increase was 42% in fine-textured soils and 30% in coarse-textured soils. Penetration resistance ranged from 0.45 MPa under forest to 4.25 MPa in old pastures, with maximum values occurring at 3–12 cm depth in pastures older than 9 years. Average total soil porosity was 58–62% under forest and 46–49% under pasture. Basic infiltration dropped from 15 cm h⁻¹ in the original forest conditions to less than 1 cm h⁻¹ in old pastures. Crude protein content and dry matter yield of the forage grass steadily decreased over time. No clear relationship between declining protein content as a function of pasture age and changes in chemical soil properties was found, but there was a high negative correlation (r=−0.81) between protein content and bulk density, reflecting the effect of soil compaction on pasture performance. After about 9–10 years of use, established grass did no longer compete successfully with invading weeds and grazing plots were abandoned. As land is not yet a scarcity in this colonisation area, degraded pastures are seldom rehabilitated.     

Effect of summer throughfall exclusion, summer drought, and winter snow cover on methane fluxes in a temperate forest soil

Soil moisture strongly controls the uptake of atmospheric methane by limiting the diffusion of methane into the soil, resulting in a negative correlation between soil moisture and methane uptake rates under most non-drought conditions. However, little is known about the effect of water stress on methane uptake in temperate forests during severe droughts. We simulated extreme summer droughts by exclusion of 168 mm (2001) and 344 mm (2002) throughfall using three translucent roofs in a mixed deciduous forest at the Harvard Forest, Massachusetts, USA. The treatment significantly increased CH₄ uptake during the first weeks of throughfall exclusion in 2001 and during most of the 2002 treatment period. Low summertime CH₄ uptake rates were found only briefly in both control and exclusion plots during a natural late summer drought, when water contents below 0.15 g cm⁻³ may have caused water stress of methanotrophs in the A horizon. Because these soils are well drained, the exclusion treatment had little effect on A horizon water content between wetting events, and the effect of water stress was smaller and more brief than was the overall treatment effect on methane diffusion. Methane consumption rates were highest in the A horizon and showed a parabolic relationship between gravimetric water content and CH₄ consumption, with maximum rate at 0.23 g H₂O g⁻¹ soil. On average, about 74% of atmospheric CH₄ was consumed in the top 4-5 cm of the mineral soil. By contrast, little or no CH₄ consumption occurred in the O horizon. Snow cover significantly reduced the uptake rate from December to March. Removal of snow enhanced CH₄ uptake by about 700-1000%, resulting in uptake rates similar to those measured during the growing season. Soil temperatures had little effect on CH₄ uptake as long as the mineral soil was not frozen, indicating strong substrate limitation of methanotrophs throughout the year. Our results suggest that the extension of snow periods may affect the annual rate of CH₄ oxidation and that summer droughts may increase the soil CH₄ sink of temperate forest soils.     

Microbial properties and soil respiration in submontane forests of Venezuelian Guyana: characteristics and response to fertilizer treatments

The distribution of vegetation types in Venezuelan Guyana (in the ‘Canaima’ National Park) represents a transitional stage in a long term process of savannization, a process considered to be conditioned by a combined chemical and intermittent drought stress. All types of woody vegetation in this environment accumulate large amounts of litter and soil organic carbon (SOC). We hypothesized that this accumulation is caused by low microbial activity. During 1 year we measured microbial biomass carbon (Cmic), microbial respiration and soil respiration of stony Oxisols (Acrohumox) at a tall, a medium and a low forest and with three chemical modifications of site conditions by the addition of NO₃⁻, Ca²⁺ and PO₄³⁻ as possible limiting elements. Due to high SOC contents, mean Cmic was 1 mg g soil⁻¹ in the mineral topsoil and 3 mg g soil⁻¹ in the forest floor. Mean microbial respiration in the mineral topsoil and the forest floor were 165 and 192 μg CO₂-C g soil⁻¹ d⁻¹, respectively. We calculated high mean metabolic quotients (qCO₂) of 200 mg CO₂-C g Cmic⁻¹ d⁻¹ in the litter layer and 166 mg CO₂-C g Cmic⁻¹ d⁻¹ in the mineral topsoil, while the Cmic-to-SOC ratios were as low as 1.0% in the litter layer and 0.8% in the mineral topsoil. Annual soil respiration was 9, 12 and 10 Mg CO₂-C ha⁻¹ yr⁻¹ in the tall, medium and low forest, respectively. CO₂ production was significantly increased by CaHPO₄ fertilization, but no consistent effects were caused by Ca²⁺ and NO₃⁻, fertilization. Our findings indicate that Cmic and microbial respiration are reduced by low nutrient concentrations and low litter and SOC quality. Reduced microbial decomposition may have contributed to SOC accumulation in these forests.     

Experimental nitrogen deposition alters the quantity and quality of soil dissolved organic carbon in an alpine meadow on the Qinghai-Tibetan Plateau

Dissolved organic matter (DOM) plays a central role in driving biogeochemical processes in soils, but little information is available on the relation of soil DOM dynamics to microbial activity. The effects of NO₃⁻ and NH₄⁺ deposition in grasslands on the amount and composition of soil DOM also remain largely unclear. In this study, a multi-form, low-dose N addition experiment was conducted in an alpine meadow on the Qinghai–Tibetan Plateau in 2007. Three N fertilizers, NH₄Cl, (NH₄)₂SO₄ and KNO₃, were applied at four rates: 0, 10, 20 and 40 kg N ha⁻¹ yr⁻¹. Soil samples from surface (0–10 cm) and subsurface layers (10–20 cm) were collected in 2011. Excitation/emission matrix fluorescence spectroscopy (EEM) was used to assess the composition and stability of soil DOM. Community-level physiological profile (CLPP, basing on the BIOLOG Ecoplate technique) was measured to evaluate the relationship between soil DOC dynamics and microbial utilization of C resources. Nitrogen (N) dose rather than N form significantly increased soil DOC contents in surface layer by 23.5%–35.1%, whereas it significantly decreased soil DOC contents in subsurface layer by 10.4%–23.8%. Continuous five-year N addition significantly increased the labile components and decreased recalcitrant components of soil DOM in surface layer, while an opposite pattern was observed in subsurface layer; however, the humification indices (HIX) of soil DOM was unaltered by various N treatments. Furthermore, N addition changed the amount and biodegradability of soil DOM through stimulating microbial metabolic activity and preferentially utilizing organic acids. These results suggest that microbial metabolic processes dominate the dynamics of soil DOC, and increasing atmospheric N deposition could be adverse to the accumulation of soil organic carbon pool in the alpine meadow on the Qinghai-Tibetan Plateau.     

Production,oxidation, emission and consumption of methane by soils: A review

Methane emission by soils results from antagonistic but correlated microbial activities. Methane is produced in the anaerobic zones of submerged soils by methanogens and is oxidised into CO₂ by methanotrophs in the aerobic zones of wetland soils and in upland soils. Methanogens and methanotrophs are ubiquitous in soils where they remain viable under unfavourable conditions. Methane transfer from the soil to the atmosphere occurs mostly through the aerenchyma of aquatic plants, but also by diffusion and as bubbles escaping from wetland soils. Methane sources are mainly wetlands. However 60 to more than 90 % of CH₄ produced in the anaerobic zones of wetlands is reoxidised in their aerobic zones (rhizosphere and oxidised soil-water interface). Methane consumption occurs in most soils and exhibits a broad range of values. Highest consumption rates or potentials are observed in soils where methanogenesis is or has been effective and where CH₄ concentration is or has been much higher than in the atmosphere (ricefields, swamps, landfills, etc.). Aerobic soils consume atmospheric CH₄ but their activities are very low and the micro-organisms involved are largely unknown. Methane emissions by cultivated or natural wetlands are expressed in mg CH₄·m^–2·h^–1 with a median lower than 10 mg CH₄·m^–2·h^–1. Methanotrophy in wetlands is most often expressed with the same unit. Methane oxidation by aerobic upland soils is rarely higher than 0.1 mg CH₄·m^–2·h^–1. Forest soils are the most active, followed by grasslands and cultivated soils. Factors that favour CH₄ emission from cultivated wetlands are mostly submersion and organic matter addition. Intermittent drainage and utilisation of the sulphate forms of N-fertilisers reduce CH₄ emission. Methane oxidation potential of upland soils is reduced by cultivation, especially by ammonium N-fertiliser application.     

A quick and sensitive method for the quantification of peroxidase activity of organic surface soil from forests

The purpose of the present study was to test the non-mutagenic compound 3,3′,5,5′-tetramethylbenzidine (TMB) as a model substrate for peroxidase in forest topsoil, as an alternative to the conventional substrate l-3,4-dihydroxyphenylalanine (l-DOPA). TMB was highly sensitive; linear absorbance changes of 0.6 were achieved within 20 min for 1000-fold diluted soil. Brief heating (denaturation) of the soil suspension gave a 34-fold reduction of TMB oxidation, indicating that the reaction measured by TMB was indeed an enzymatic reaction. TMB oxidation showed a narrow peak at pH 4.4. A proportional decrease in peroxidase activity, when the soil suspension was diluted, demonstrated that TMB estimates of peroxidase activity are directly comparable when corrected for differences in sample size. Oxidation of TMB was slow in the absence of H₂O₂ suggesting that TMB is a poor substrate for phenol oxidases. TMB oxidation was tested in nine different forest topsoils. The peroxidase activity, when normalised to the amount of soil organic matter, ranged from 1.4±0.1 Δabs₄₅₀ h^?1 mg^?1 to 34.9±4.3 Δabs₄₅₀ h^?1 mg^?1. In comparison, l-DOPA oxidation by soil peroxidases and commercial peroxidases gave inconsistent results, suggesting that one should be cautious when using l-DOPA as a soil peroxidase substrate. The high sensitivity of TMB, compared to l-DOPA, and the low interference from phenol oxidase and humic substances suggest that TMB is a better substrate than l-DOPA for estimation of peroxidase activity of forest topsoil.     

Agricultural factors affecting methane oxidation in arable soil

CH₄ oxidation activity in a sandy soil (Ardoyen) and agricultural practices affecting this oxidation were studied under laboratory conditions. CH₄ oxidation in the soil proved to be a biological process. The instantaneous rate of CH₄ consumption was in the order of 800 mol CH₄ kg^–1 day^–1 (13 mg CH₄ kg^–1 day^–1) provided the soil was treated with ca. 4.0 mmol CH₄ kg^–1 soil. Upon repeated supplies of a higher dose of CH₄, the oxidation was accelerated to a rate of at least 198 mg CH₄ kg^–1 day^–1. Addition of the plant-growth promoting rhizopseudomonad strains Pseudomonas aeruginosa 7NSK2 and Pseudomonas fluorescens ANP15 significantly decreased the CH₄ oxidation by 20 to 30% during a 5-day incubation. However, with further incubation this suppression was no longer detectable. Growing maize plants prevented the suppression of CH₄ oxidation. The numbers of methanotrophic bacteria and fungi increased significantly after the addition of CH₄, but there were no significant shifts in the population of total bacteria and fluorescent pseudomonads. Drying and rewetting of soil for at least 1 day significantly reduced the activity of the indigenous methanotrophs. Upon rewetting, their activity was regained after a lag phase of about 3 days. The herbicide dichlorophenoxy acetic acid (2,4-D) had a strong negative effect on CH₄ oxidation. The application of 5 ppm increased the time for CH₄ removal; at concentrations above 25 ppm 2,4-D CH₄–oxidizing activity was completely hampered. After 3 days of delay, only the treatments with below 25 ppm 2,4-D showed recovery of CH₄–oxidizing activity. This finding suggests that it can be important to include a CH₄–removal bioassay in ecotoxicology studies of the side effects of pesticides. Changes in the native soil pH also affected the CH₄–oxidizing capacity. Permanent inhibition occurred when the soil pH was altered by 2 pH units, and partial inhibition by 1 pH unit change. A rather narrow pH range (5.9–7.7) appeared to allow CH₄ oxidation. Soils pre-incubated with NH ₄ ⁺ had a lower CH₄–removal capacity. Moreover, the nitrification inhibitor 2-chloro-6-trichloromethyl pyridine (nitrapyrin) strongly inhibited CH₄ oxidation. Probably methanotrophs rather than nitrifying microorganisms are mainly responsible for CH₄ removal in the soil studied. It appears that the causal methanotrophs are remarkably sensitive to soil environmental disturbances.