Potassium application reduces methane emission from a flooded field planted to rice

In a field study, potassium (K) applied as muriate of potash (MOP) significantly reduced methane (CH₄) emission from a flooded alluvial soil planted to rice. Cumulative emission was highest in control plots (125.34 kg CH₄ ha⁻¹), while the lowest emission was recorded in field plots receiving 30 kg K ha⁻¹ (63.81 kg CH₄ ha⁻¹), with a 49% reduction in CH₄ emission. Potassium application prevented a drop in the redox potential and reduced the contents of active reducing substances and Fe²⁺ content in the rhizosphere soil. Potassium amendment also inhibited methanogenic bacteria and stimulated methanotrophic bacterial population. Results suggest that, apart form producing higher plant biomass (both above- and underground) and grain yield, K amendment can effectively reduce CH₄ emission from flooded soil and could be developed into an effective mitigation option, especially in K-deficient soils.     

Methane and water soluble iron production under controlled soil pH and redox conditions

Abstract

When a soil is flooded, iron (Fe) reduction and methane (CH₄) production occurred in sequence as predicted by thermodynamics. The dissolution and precipitation of Fe reflected both soil pH and soil redox potential (Eh). The objective of our experiment was to determine both CH₄ production and Fe reduction as measured by Fe in solution in a flooded paddy soil over a wide range of closely controlled pH and Eh conditions. The greatest release of CH₄ gas occurred at neutral soil pH in combination with low soil redox potential (‐250 mV). Production of CH₄ decreased when soil pH was lowered in combination with an increase in the soil redox potential above ‐250 mV. Highest concentration of ferrous‐iron (Fe²⁺) under reducing conditions occurred when soil pH was lowered. Thus Fe reduction influenced CH4 formation in the flooded paddy soil. Results indicated that CH₄ production was inhibited by the process of ferric‐iron (Fe³⁺) reduction.     

Potential contribution of Lumbricus terrestris L. to carbon dioxide, methane and nitrous oxide fluxes from a forest soil

 Potential effects of earthworms (Lumbricus terrestris L.) inoculated into soil on fluxes of CO₂, CH₄ and N₂O were investigated for an untreated and a limed soil under beech in open topsoil columns under field conditions for 120 days. Gas fluxes from L. terrestris, beech litter and mineral soil from soil columns were measured separately in jars at 17  °C. The inoculation with L. terrestris and the application of lime had no effect on cumulative CO₂ emissions from soil. During the first 3–4 weeks earthworms significantly (P<0.05) increased CO₂ emissions by 16% to 28%. In contrast, significantly lower (P<0.05) CO₂ emission rates were measured after 11 weeks. The data suggest that earthworm activity was high during the first weeks due to the creation of burrows and incorporation of beech litter into the mineral soil. Low cumulative CH₄ oxidation rates were found in all soil columns as a result of CH₄ production and oxidation processes. L. terrestris with fresh feces and the beech litter produced CH₄ during the laboratory incubation, whereas the mineral soil oxidised atmospheric CH₄. Inoculation with L. terrestris led to a significant reduction (P<0.02) in the CH₄ oxidation rate of soil, i.e. 53% reduction. Liming had no effect on cumulative CH₄ oxidation rates of soil columns and on CH₄ fluxes during the laboratory incubation. L. terrestris significantly increased (P<0.001) cumulative N₂O emissions of unlimed soil columns by 57%. The separate incubation of L. terrestris with fresh feces resulted in rather high N₂O emissions, but the rate strongly decreased from 54 to 2 μg N kg^–1 (dry weight) h^–1 during the 100 h of incubation. Liming had a marked effect on N₂O formation and significantly (P<0.001) reduced cumulative N₂O emissions by 34%. Although the interaction of liming and L. terrestris was not significant, N₂O emissions of limed soil columns with L. terrestris were 8% lower than those of the control. Received: 2 September 1999     

Methane emission from flooded rice fields under irrigated conditions

In a study on CH₄ emission from flooded rice fields under irrigated conditions, fields planted with rice emitted more methane than unplanted fields. The CH₄ efflux in planted plots varied with the rice variety and growth stage and ranged from 4 to 26 mg h^-1m^-2. During the reproductive stage of the rice plants, CH₄ emission was high and the oxidation power of rice roots, in terms of -naphthylamine oxidation, was very low. The CH₄ emission reached a maximum at midday and declined to minimum levels at midnight, irrespective of the rice variety. The peak CH₄ emission at midday was associated with higher solar radiation and higher soil/water temperature.     

Short- and long-term effects of nitrogen fertilization on methane oxidation in three Swedish forest soils

 Under normal conditions, CH₄, one of the most important greenhouse gases, is subject to biological oxidation in forest soils. However, this process can be negatively affected by N amendment. The reported experiment was conducted in order to study the short- and long-term effects of N amendment on CH₄ oxidation in pine (Pinus sylvestris L.) forest soils. Soil samples were taken from three experimental sites, two of which had been amended with N once, over 20 years earlier, while the third had been amended 3 weeks earlier. The soil samples were incubated fresh at 15  °C at ambient CH₄ concentrations (ca. 1.8 ppmv CH₄). The variation in CH₄-turnover rates was high within the treatments: CH₄ was produced [up to 22.6 pmol CH₄ g dry wt. soil^–1 h^–1] in samples from the recently amended site, whereas it was consumed at high rates (up to 431 pmol CH₄ g dry wt. soil^–1 h^–1) in samples from the plot that had received the highest N amendment 27 years before sampling. Although no significant differences were found between N treatments, in the oldest plots there was a correlation between consumption of atmospheric CH₄ and the total C content at a depth of 7.5–15 cm in the mineral soil (r ²=0.74). This indicates that in the long-term, increased C retention in forest soils following N amendment could lead to increased CH₄ oxidation. Received: 3 September 1997     

Reduced net atmospheric CH4 consumption is a sustained response to elevated CO2 in a temperate forest

We compared, from 2004 through 2006, rates of soil–atmosphere CH₄ exchange at permanently established sampling sites in a temperate forest exposed to ambient (control plots; ∼380 μL L⁻¹) or elevated (ambient + 200 μL L⁻¹) CO₂ since August 1996. A total of 880 observations showed net atmospheric CH₄ consumption (flux from the atmosphere to the soil) from all static chambers most of the time at rates varying from 0.02 mg m⁻² day⁻¹ to 4.5 mg m⁻² day⁻¹. However, we infrequently found net CH₄ production (flux from the soil to the atmosphere) at lower rates, 0.01 mg m⁻² day⁻¹ to 0.08 mg m⁻² day⁻¹. For the entire study, the mean (±SEM) rate of net CH₄ consumption in control plots was higher than the mean for CO₂-enriched plots, 0.55 (0.03) versus 0.51 (0.03) mg m⁻² day⁻¹. Annual rates of 184, 196, and 197 mg m⁻² for net CH₄ consumption at control plots during the three calendar years of this study were 19, 10, and 8% higher than comparable values for CO₂ enriched plots. Differences between treatments were significant in 2004 and 2005 and nearly significant in 2006. Volumetric soil water content was consistently higher at CO₂-enriched sites and a mixed-effects model identified a significant soil moisture x CO₂ interaction on net atmospheric CH₄ consumption. Increased soil moisture at CO₂-enriched sites likely increases diffusional resistance of surface soils and the frequency of anaerobic microsites supporting methanogenesis, resulting in reduced rates of net atmospheric CH₄ consumption. Our study extends our observations of reduced net atmospheric CH₄ consumption at CO₂-enriched plots to nearly five continuous years, suggesting that this is likely a sustained negative feedback to increasing atmospheric CO₂ at this site.     

Increased moisture and methanogenesis contribute to reduced methane oxidation in elevated CO2 soils

Awareness of global warming has stimulated research on environmental controls of soil methane (CH₄) consumption and the effects of increasing atmospheric carbon dioxide (CO₂) on the terrestrial CH₄ sink. In this study, factors impacting soil CH₄ consumption were investigated using laboratory incubations of soils collected at the Free Air Carbon Transfer and Storage I site in the Duke Forest, NC, where plots have been exposed to ambient (370 μL L⁻¹) or elevated (ambient + 200 μL L⁻¹) CO₂ since August 1996. Over 1 year, nearly 90% of the 360 incubations showed net CH₄ consumption, confirming that CH₄-oxidizing (methanotrophic) bacteria were active. Soil moisture was significantly (p < 0.01) higher in the 25–30 cm layer of elevated CO₂ soils over the length of the study, but soil moisture was equal between CO₂ treatments in shallower soils. The increased soil moisture corresponded to decreased net CH₄ oxidation, as elevated CO₂ soils also oxidized 70% less CH₄ at the 25–30 cm depth compared to ambient CO₂ soils, while CH₄ consumption was equal between treatments in shallower soils. Soil moisture content predicted (p < 0.05) CH₄ consumption in upper layers of ambient CO₂ soils, but this relationship was not significant in elevated CO₂ soils at any depth, suggesting that environmental factors in addition to moisture were influencing net CH₄ oxidation under elevated CO₂. More than 6% of the activity assays showed net CH₄ production, and of these, 80% contained soils from elevated CO₂ plots. In addition, more than 50% of the CH₄-producing flasks from elevated CO₂ sites contained deeper (25–30 cm) soils. These results indicate that subsurface (25 cm+) CH₄ production contributes to decreased net CH₄ consumption under elevated CO₂ in otherwise aerobic soils.     

Methane production in a flooded soil in response to elevated atmospheric carbon dioxide concentrations

 CH₄ production in a flooded soil as affected by elevated atmospheric CO₂ was quantified in a laboratory incubation study. CH₄ production in the flooded soil increased by 19.6%, 28.2%, and 33.4% after a 2-week incubation and by 38.2%, 62.4%, and 43.0% after a 3-week incubation under atmospheres of 498, 820, and 1050 μl l^–1 CO₂, respectively, over that in soil under the ambient CO₂ concentration. CH₄ production in slurry under 690, 920, and 1150 μl l^–1 CO₂ increased by 2.7%, 5.5%, and 5.0%, respectively, after a 3-day incubation, and by 6.7%, 12.8%, and 5.4%, respectively, after a 6-day incubation over that in slurry under the ambient CO₂ concentration. The increase in CH₄ production in the soil slurry under elevated CO₂ concentrations in a N₂ atmosphere was more pronounced than that under elevated CO₂ concentrations in air. These data suggested that elevated atmospheric CO₂ concentrations could promote methanogenic activity in flooded soil. Received: 2 March 1998     

Effects of rice‐straw and phosphorus application on production and emission of methane from tropical rice soil

Rice‐straw amendment increased methane production by 3‐fold over that of unamended control. Application of P as single superphosphate at 100 μg (g soil)^–1 inhibited methane (CH₄) production distinctly in flooded alluvial rice soil, in the absence more than in the presence of rice straw. CH₄ emission from rice plants (cv. IR72) from alluvial soil treated with single superphosphate as basal application, in the presence and absence of rice straw, and held under non‐flooded and flooded conditions showed distinct variations. CH₄ emission from non‐flooded soil amended with rice straw was high and almost similar to that of flooded soil without rice‐straw amendment. The cumulative CH₄ efflux was highest (1041 mg pot^–1) in rice‐straw‐amended flooded soil. Appreciable methanogenic reactions in rice‐straw‐amended soils were evident under both flooded and non‐flooded conditions. Rice‐straw application substantially altered the balance between total aerobic and anaerobic microorganisms even in non‐flooded soil. The mitigating effects of single‐superphosphate application or low‐moisture regime on CH₄ production and emission were almost nullified due to enhanced activities of methanogenic archaea in the presence of rice straw.     

Methane emission from paddy fields in Taiwan

 In order to investigate the effect of environmental conditions on CH₄ emission from paddy fields in Taiwan, four locations, two cropping seasons and two irrigation systems were studied. CH₄ emission was high at the active tillering and the booting stages in the first cropping season, whereas it was low at the transplanting and the ripening stages with an intermittent irrigation system. CH₄ emission was high at the transplanting stage in the second cropping season, and decreased gradually during rice cultivation. Daily temperature and light intensity increased gradually during rice growth in the first cropping season (February–June), while it was reversed in the second cropping season (August–December). The seasonal CH₄ emission from paddy fields ranged from 1.73 to 11.70 g m^–2, and from 10.54 to 39.50 g m^–2 in the first and second cropping seasons, respectively. The seasonal CH₄ emission in the second cropping season was higher than that in the first cropping season in all test fields. The seasonal CH₄ emission was 32.65 mg m^–2 in the first cropping season of the National Taiwan University paddy field with continuous flooding, and it was 28.85 mg m^–2 in the second cropping season. The annual CH₄ emission ranged from 12.3 to 49.3 g m^–2 with an intermittent irrigation system, and the value was 61.5 g m^–2 with a continuous flooding treatment. The annual CH₄ emission from paddy fields was estimated to be 0.034 Tg in 1997 from 364,212 ha of paddy fields with an intermittent irrigation system, which was less than the 0.241 Tg calculated by the IPCC method with a continuous flooding treatment Received: 23 February 2000     

Influence of green manuring with Sesbania rostrata and Aeschynomene afraspera or urea on dynamics and uptake of nutrients by wetland rice

Nutrient concentrations in the soil and crop uptake from incorporated green manure and urea in flooded rice was studied in field experiments. Release of plant-available nitrogen (NH₄ ⁺-N) from green manure was slightly delayed compared with that from prilled urea (PU) because Sesbania rostrata L. and Aeschynomene afraspera L. released the N gradually after their decomposition, whereas N became available immediately after PU application. Exchangeable NH₄ ⁺-N concentration in soil peaked at 163 mg kg^–1 in the transplanted rice (TPR) and 198 mg kg^—1 in broadcast-seeded rice (BSR) at 0 and 1 week after PU application. Broadcast-seeded rice depleted NH₄ ⁺-N faster than did TPR because of the crop‘s vigorous growth in the former during the early stage. Soil solution NH₄ ⁺-N followed a similar trend to that of soil NH₄ ⁺-N. Incorporation of S. rostrata and A. afraspera increased the concentration of P, K⁺, Fe²⁺ and Mn²⁺ in soil solution more than did the application of PU. However, zinc concentration decreased in all treatments. Both PU and green manure increased the N status of the rice plants and enhanced the uptake of P, K, Fe, Mn and Zn by the rice crop. This suggests that application of green manures improves the uptake of these nutrients by the crop. The highest apparent N recovery was obtained with PU followed by green manure. Received: 11 November 1996     

Different response of silicate fertilizer having electron acceptors on methane emission in rice paddy soil under green manuring

Cultivation of green manure plants during the fallow season in rice paddy soil has been strongly recommended to improve soil properties. However, green manuring may impact greenhouse gas emission, methane (hereafter, CH₄) in particular, under the flooded rice cultivation and thus, application of chemical amendments being electron acceptors can be an effective mitigation strategy to reduce CH₄ emissions in irrigated rice (Oryza sativa L.) field amended with green manure. To investigate the effect of iron (Fe) slag silicate fertilizer (hereafter, silicate fertilizer), which was effective in reducing CH₄ emission and increasing rice productivity, in green manure-amended paddy soil, the aboveground biomass of Chinese milk vetch (hereafter, vetch) was added at rates of 0, 10, 20, and 40 Mg (fresh weight) ha⁻¹ before the application of silicate fertilizer, which was added at rates of 0 and 2.3 Mg ha⁻¹. Silicate fertilization reduced the seasonal CH₄ flux by ca. 14.5% and increased rice yield by ca. 15.7% in the control (no vetch application) treatment. However, CH₄ production was increased by silicate fertilization in vetch-treated soil particularly at the initial rice growing stage, which was probably due to the enhanced decomposition of added organic matters by the silicate liming effect. In conclusion, silicate fertilization is not effective in reducing CH₄ production in green manure-amended rice paddy soils and its use should be properly controlled.     

Influence of N and non-N salts on atmospheric methane oxidation by upland boreal forest and tundra soils

 The short-term (24 h) and medium-term (30 day) influence of N salts (NH₄Cl, NaNO₃ and NaNO₂) and a non-N salt (NaCl) on first-order rate constants, k (h^–1) and thresholds (C_Th) for atmospheric CH₄ oxidation by homogenized composites of upland boreal forest and tundra soils was assessed at salt additions ranging to 20 μmol g^–1 dry weight (dw) soil. Additions of NH₄Cl, NaNO₃ and NaCl to 0.5 μmol g^–1 dw soil did not significantly decrease k relative to watered controls in the short term. Higher concentrations significantly reduced k, with the degree of inhibition increasing with increasing dose. Similar doses of NH₄Cl and NaCl gave comparable decreases in k relative to controls and both soils showed low native concentrations of NH₄ ⁺-N (≤1 μmol g^–1dw soil), suggesting that the reduction in k was due primarily to a salt influence rather than competitive inhibition of CH₄ oxidation by exogenous NH₄ ⁺-N or NH₄ ⁺-N released through cation exchange. The decrease in k was consistently less for NaNO₃ than for NH₄Cl and NaCl at similar doses, pointing to a strong inhibitory effect of the Cl^– counter-anion. Thresholds for CH₄ oxidation were less sensitive to salt addition than k for these three salts, as significant increases in C_Th relative to controls were only observed at concentrations ≥1.0 μmol g^–1 dw soil. Both soils were more sensitive to NaNO₂ than to other salts in the short term, showing a significant decrease in k at an addition of 0.25 μmol NaNO₂ g^–1 dw soil that was clearly attributable to NO₂ ^–. Soils showed no recovery from NaCl, NH₄ ⁺-N or NaNO₃ addition with respect to atmospheric CH₄ oxidation after 30 days. However, soils amended with NaNO₂ to 1.0 μmol NaNO₂ g^–1 dw showed values of k that were not significantly different from controls. Recovery of CH₄-oxidizing ability was due to complete oxidation of NO₂ ^–-N to NO₃ ^–-N. Analysis of soil concentrations of N salts necessary to inhibit atmospheric CH₄ oxidation and regional rates of N deposition suggest that N deposition will not decrease the future sink strength of upland high-latitude soils in the atmospheric CH₄ budget. Received: 30 April 1999     

Methane production in unamended and rice-straw-amended soil at different moisture levels

 CH₄ production in an alluvial soil, unamended or amended with rice straw (1% w/w), was examined under nonflooded [–1.5 MPa, –0.01 MPa and 0 MPa (saturated) and flooded (1 : 1.25 soil to water ratio)] conditions during a 40-day incubation in closed Vacutainer tubes. CH₄ production was negligible at –1.5 MPa, but increased with an increase in the moisture level. Addition of rice straw distinctly increased CH₄ production in the soil at all moisture levels including –1.5 MPa. Evidence, in terms of the drop in redox potential and Fe²⁺ accumulated, suggested that the addition of rice straw hastened the reduction of the soil, even under nonflooded conditions; thus its addition stimulated even the nonflooded soil to produce CH₄ in substantial amounts. Our results indicate that many currently unidentified sources of CH₄, possibly including organic-amended nonflooded soils, may make a significant contribution to the global CH₄ budget. Received: 10 July 1997     

Short-term effects of nitrogen on methane oxidation in soils

 The short-term effects of N addition on CH₄ oxidation were studied in two soils. Both sites are unfertilized, one has been under long-term arable rotation, the other is a grassland that has been cut for hay for the past 125 years. The sites showed clear differences in their capacity to oxidise CH₄, the arable soil oxidised CH₄ at a rate of 0.013 μg CH₄ kg^–1 h^–1 and the grassland soil approximately an order of magnitude quicker. In both sites the addition of (NH₄)₂SO₄ caused an immediate reduction in the rate of atmospheric CH₄ oxidation approximately in inverse proportion to the amount of NH₄ ⁺ added. The addition of KNO₃ caused no change in the rate of CH₄ oxidation in the arable soil, but in the grassland soil after 9 days the rate of CH₄ oxidation had decreased from 0.22 μg CH₄ kg^–1 h^–1 to 0.13 μg CH₄ kg^–1 h^–1 in soil treated with the equivalent of 192 kg N ha^–1. A ¹⁵N isotopic dilution technique was used to investigate the role of nitrifiers in regulating CH₄ oxidation. The arable soil showed a low rate of gross N mineralisation (0.67 mg N kg^–1 day^–1), but a relatively high proportion of the mineralised N was nitrified. The grassland soil had a high rate of gross N mineralisation (18.28 mg N kg^–1 day^–1), but negligible nitrification activity. It is hypothesised that since there was virtually no nitrification in the grassland soil then CH₄ oxidation at this site must be methanotroph mediated. Received: 31 October 1997     

Effects of 4-amino 1,2,4-triazole, dicyandiamide and encapsulated calcium carbide on nitrification inhibition in a subtropical soil under upland and flooded conditions

 Nitrification inhibition of soil and applied fertilizer N is desirable as the accumulation of nitrates in soils in excess of plant needs leads to enhanced N losses and reduced fertilizer N-use efficiency. In a growth chamber experiment, we studied the effects of two commercial nitrification inhibitors (NIs), 4-amino 1,2,4-triazole (ATC) and dicyandiamide (DCD), and a commonly available and economical material, encapsulated calcium carbide (CaC₂) (ECC) on the nitrification of soil and applied NH₄ ⁺-N in a semiarid subtropical Tolewal sandy loam soil under upland [60% water-filled pore space (WFPS)] and flooded conditions (120% WFPS). Nitrification of the applied 100 mg NH₄ ⁺-N kg^–1 soil under upland conditions was retarded most effectively (93%) by ECC for up to 10 days of incubation, whereas for longer periods, ATC was more effective. After 20 days, only 16% of applied NH₄ ⁺-N was nitrified with ATC as compared to 37% with DCD and 98% with ECC. Under flooded soil conditions, nitrates resulting from nitrification quickly disappeared due to denitrification, resulting in a tremendous loss of fertilizer N (up to 70% of N applied without a NI). Based on four indicators of inhibitor effectiveness, namely, concentration of NH₄ ⁺-N and NO₃ ^–-N, percent nitrification inhibition, ratio of NH₄ ⁺-N/NO₃ ^–-N, and total mineral N, ECC showed the highest relative efficiency throughout the 20-day incubation under flooded soil conditions. At the end of the 20-day incubation, 96%, 58% and 38% of applied NH₄ ⁺-N was still present in the soil where ECC, ATC and DCD were used, respectively. Consequently, nitrification inhibition of applied fertilizer N in both arable crops and flooded rice systems could tremendously minimize N losses and help enhance fertilizer N-use efficiency. These results suggest that for reducing the nitrification rate and resultant N losses in flooded soil systems (e.g. rice lowlands), ECC is more effective than costly commercial NIs. Received: 25 May 2000     

除草剂对不同种植年限柑橘园土壤氮转化过程及温室气体排放的影响

为探讨除草剂施用对柑橘园土壤氮转化及温室气体排放的影响,在实验室培养条件下,研究了0年(林地)、种植10年和30年的柑橘园土壤中分别添加除草剂草甘膦和丁草胺后,尿素态氮含量、硝化和反硝化作用以及温室气体排放的变化。研究结果表明,橘园土壤中尿素第1 d的水解率、氮肥硝化率、反硝化作用损失总量以及N_2O和CO_2排放量显著高于林地土壤(P0.05)。与10年橘园土壤相比,30年橘园土壤显著增加了尿素的水解速率、氮肥硝化率和CO_2排放量(P0.05),但二者的反硝化损失量没有显著差异。施用草甘膦和丁草胺都显著促进了林地土壤的尿素水解(P0.05),第1 d尿素态氮含量分别降低11.20%和12.43%;但对3种土壤氮肥的硝化率均没有明显影响。施用丁草胺显著降低了林地土壤的CO_2排放量(P0.05),对两种橘园土壤的CO_2排放没有明显影响,但明显增加了两种橘园土壤的N_2O排放总量(P0.05),分别比不施除草剂增加56.27%和85.41%;施用草甘膦对3种土壤的N_2O和CO_2排放均没有明显影响。可见,草甘膦和丁草胺的施用不会对柑橘园土壤的氮转化过程产生影响,但丁草胺显著增加了柑橘园土壤的N_2O排放。     

Abundance and community composition of methanotrophs in a Chinese paddy soil under long-term fertilization practices

Background, aim, and scope  As the second most important greenhouse gas, methane (CH₄) is produced from many sources such as paddy fields. Methane-oxidizing bacteria (methanotrophs) consume CH₄ in paddy soil and, therefore, reduce CH₄ emission to the atmosphere. In order to estimate the contribution of paddy fields as a source of CH₄, it is important to monitor the effects of fertilizer applications on the shifts of soil methanotrophs, which are targets in strategies to combat global climate change. In this study, real-time polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA and pmoA genes, respectively, were used to analyze the soil methanotrophic abundance and community diversity under four fertilization treatments: urea (N), urea and potassium chloride (NK), urea, superphosphate, and potassium chloride (NPK), and urea, superphosphate, potassium chloride, and crop residues (NPK+C), compared to an untreated control (CON). The objective of this study was to examine whether soil methanotrophs responded to the long-term, different fertilizer regimes by using a combination of quantitative and qualitative molecular approaches. Materials and methods  Soil samples were collected from the Taoyuan Experimental Station of Agro-ecosystem Observation at Changde (28°55′ N, 111°26′ E), central Hunan Province of China, in July 2006. Soil DNAs were extracted from the samples, then the 16S rRNA genes were quantified by real-time PCR and the pmoA genes were amplified via general PCR followed by DGGE, cloning, sequencing, and phylogenetic analysis. The community diversity indices were assessed through the DGGE profile. Results  Except for NPK, other treatments of N, NK, and NPK+C showed significantly higher copy numbers of type I methanotrophs (7.0–9.6 × 10⁷) than CON (5.1 × 10⁷). The copy numbers of type II methanotrophs were significantly higher in NPK+C (2.8 × 10⁸) and NK (2.5 × 10⁸) treatments than in CON (1.4 × 10⁸). Moreover, the ratio of type II to type I methanotrophic copy numbers ranged from 1.88 to 3.32, indicating that the type II methanotrophs dominated in all treatments. Cluster analyses based on the DGGE profile showed that the methanotrophic community in NPK+C might respond more sensitively to the environmental variation. Phylogenetic analysis showed that 81% of the obtained pmoA sequences were classified as type I methanotrophs. Furthermore, the type I-affiliated sequences were related to Methylobacter, Methylomicrobium, Methylomonas, and some uncultured methanotrophic clones, and those type II-like sequences were affiliated with Methylocystis and Methylosinus genera. Discussion  There was an inhibitory effect on the methanotrophic abundance in the N and a stimulating effect in the NK and NPK+C treatments, respectively. During the rice-growing season, the type II methanotrophs might be more profited from such a coexistence of low O₂ and high CH₄ concentration environment than the type I methanotrophs. However, type I methanotrophs seemed to be more frequently detected. The relatively complex diversity pattern in the NPK+C treatment might result from the strong CH₄ production. Conclusions  Long-term fertilization regimes can both affect the abundance and the composition of the type I and type II methanotrophs. The inhibited effects on methanotrophic abundance were found in the N treatment, compared to the stimulated effects from the NK and NPK+C treatments. The fertilizers of nitrogen, potassium, and the crop residues could be important factors controlling the abundance and community composition of the methanotrophs in the paddy soil. Recommendations and perspectives  Methanotrophs are a fascinating group of microorganisms playing an important role in the biogeochemical carbon cycle and in the control of global climate change. However, it is still a challenge for the cultivation of the methanotrophs, although three isolates were obtained in the extreme environments very recently. Therefore, future studies will be undoubtedly conducted via molecular techniques just like the applications in this study.     

Factors influencing nitrous oxide and methane emissions from minerotrophic fens in northeast Germany

 At two field sites representing northeastern German minerotrophic fens (Rhin-Havelluch, a shallow peat site; Gumnitz, a partially drained peat site) the influence of different factors (N fertilization, groundwater table, temperature) on N₂O and CH₄ emissions was investigated. The degraded fens were sources or sinks of the radiatively active trace gases investigated. The gas fluxes measured were much higher than those found in other terrestrical ecosystems such as forests. Lowering the groundwater table increased the release of N₂O and the oxidation of CH₄. High CH₄ emission rates occurred when the groundwater tables and soil temperatures were high (>12  °C). N fertilization stimulated the release of N₂O only when application rates were very high (480 kg N ha^–1). A moderate N supply (60 or 120 kg N ha^–1) hardly increased the release of N₂O in spite of high soluble soil NO₃ ^– contents. Received: 31 October 1997     

Nitrification and denitrification in the rhizosphere of rice: the detection of processes by a new multi-channel electrode

 N turnover in flooded rice soils is characterized by a tight coupling between nitrification and denitrification. Nitrification is restricted to the millimetre-thin oxic surface layer while denitrification occurs in the adjacent anoxic soil. However, in planted rice soil O₂ released from the rice roots may also support nitrification within the otherwise anoxic bulk soil. To locate root-associated nitrification and denitrification we constructed a new multi-channel microelectrode that measures NH₄ ⁺, NO₂ ^–, and NO₃ ^– at the same point. Unfertilized, unplanted rice microcosms developed an oxic-anoxic interface with nitrification taking place above and denitrification below ca. 1 mm depth. In unfertilized microcosms with rice plants, NH₄ ⁺, NO₂ ^– and NO₃ ^– could not be detected in the rhizosphere. Assimilation by the rice roots reduced the available N to a level where nitrification and denitrification virtually could not occur. However, a few hours after injecting (NH₄)₂HPO₄ or urea, a high nitrification activity could be detected in the surface layer of planted microcosms and in a depth of 20–30 mm in the rooted soil. O₂ concentrations of up to 150 μM were measured at the same depth, indicating O₂ release from the rice roots. Nitrification occurred at a distance of 0–2 mm from the surface around individual roots, and denitrification occurred at a distance of 1.5–5.0 mm. Addition of urea to the floodwater of planted rice microcosms stimulated nitrification. Transpiration of the rice plants caused percolation of water resulting in a mass flow of NH₄ ⁺ towards the roots, thus supporting nitrification. Received: 23 July 1999