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1.
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.  相似文献   

2.
It has been known that nitrogenous fertilizers can either stimulate or inhibit methane oxidation in soils. The mechanism, however, remains unclear. Here we conducted laboratory incubation experiments to evaluate the effects of ammonium versus nitrate amendment on CH4 oxidation in a rice field soil. The results showed that both N forms stimulated CH4 oxidation. But nitrate stimulated CH4 oxidation to a greater extent than ammonium per unit N base. The 16S rRNA genes and the pmoA genes were analyzed to determine the dynamics of total bacterial and methanotrophic populations, respectively. The methanotrophic community consisted of type I and type II methanotrophs and was dominated by type I group after two weeks of incubation. Nitrate promoted both types of methanotrophs, but ammonium promoted only type I. DNA-based stable isotope probing confirmed that ammonium stimulated the incorporation of 13CH4 into type I methanotrophs but not type II, while nitrate caused almost homogenous distribution of 13CH4 in type I and type II methanotrophs. Our study suggests that nitrate can promote CH4 oxidation more significantly than ammonium and is probably a better N source for both types of methanotrophs in rice field soil. More investigations, e.g. using 15N labeling, are necessary to elucidate this possibility.  相似文献   

3.
The abundance, activity, and temperature response of aerobic methane-oxidizing bacteria were studied in permafrost-affected tundra soils of northeast Siberia. The soils were characterized by both a high accumulation of organic matter at the surface and high methane concentrations in the water-saturated soils. The methane oxidation rates of up to 835 nmol CH4 h−1 g−1 in the surface soils were similar to the highest values reported so far for natural wetland soils worldwide. The temperature response of methane oxidation was measured during short incubations and revealed maximum rates between 22 °C and 28 °C. The active methanotrophic community was characterized by its phospholipid fatty acid (PLFA) concentrations and with stable isotope probing (SIP). Concentrations of 16:1ω8 and 18:1ω8 PLFAs, specific to methanotrophic bacteria, correlated significantly with the potential methane oxidation rates. In all soils, distinct 16:1 PLFAs were dominant, indicating a predominance of type I methanotrophs. However, long-term incubation of soil samples at 0 °C and 22 °C demonstrated a shift in the composition of the active community with rising temperatures. At 0 °C, only the concentrations of 16:1 PLFAs increased and those of 18:1 PLFAs decreased, whereas the opposite was true at 22 °C. Similarly, SIP with 13CH4 showed a temperature-dependent pattern. When the soils were incubated at 0 °C, most of the incorporated label (83%) was found in 16:1 PLFAs and only 2% in 18:1 PLFAs. In soils incubated at 22 °C, almost equal amounts of 13C label were incorporated into 16:1 PLFAs and 18:1 PLFAs (33% and 36%, respectively). We concluded that the highly active methane-oxidizing community in cold permafrost-affected soils was dominated by type I methanotrophs under in situ conditions. However, rising temperatures, as predicted for the future, seem to increase the importance of type II methanotrophs, which may affect methane cycling in northern wetlands.  相似文献   

4.
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.  相似文献   

5.
Emissions of N2O and CH4 and CH4 oxidation rates were measured from Lolium perenne swards in a short-term study under ambient (36 Pa) and elevated (60 Pa) atmospheric CO2 at the Free Air Carbon dioxide Enrichment experiment, Eschikon, Switzerland. Elevated pCO2 increased (P<0.05) N2O emissions from high N fertilised (11.2 g N m−2) swards by 69%, but had no significant effect on net emissions of CH4. Application of 13C-CH4 (11 μl l−1; 11 at.% excess 13C) to closed chamber headspaces in microplots enabled determination of rates of 13C-CH4 oxidation even when net CH4 fluxes from main plots were positive. We found a significant interaction between fertiliser application rate and atmospheric pCO2 on 13C-CH4 oxidation rates that was attributed to differences in gross nitrification rates and C and N availability. CH4 oxidation was slower and thought to be temporarily inhibited in the high N ambient pCO2 sward. The most rapid CH4 oxidation of 14.6 μg 13C-CH4 m−2 h−1 was measured in the high fertilised elevated pCO2 sward, and we concluded that either elevated pCO2 had a stimulatory effect on CH4 oxidation or inhibition of oxidation following fertiliser application was lowered under elevated pCO2. Application of 14NH415NO3 and 15NH415NO3 (10 at.% excess 15N) to different replicates enabled determination of the respective contributions of nitrification and denitrification to N2O emissions. Inhibition of CH4 oxidation in the high fertilised ambient pCO2 sward, due to competition between NH3 and CH4 for methane monooxygenase enzymes or toxic effects of NH2OH or NO2− produced during nitrification, was hypothesised to increase gross nitrification (12.0 mg N kg dry soil−1) and N2O emissions during nitrification (327 mg 15N-N2O m−2 over 11 d). Our results indicate that increasing atmospheric concentrations of CO2 may increase emissions of N2O by denitrification, lower nitrification rates and either increase or decrease the ability of soil to act as a sink for atmospheric CH4 depending on fertiliser management.  相似文献   

6.
Emission of N2O and CH4 oxidation rates were measured from soils of contrasting (30-75%) water-filled pore space (WFPS). Oxidation rates of 13C-CH4 were determined after application of 10 μl 13C-CH4 l−1 (10 at. % excess 13C) to soil headspace and comparisons made with estimates from changes in net CH4 emission in these treatments and under ambient CH4 where no 13C-CH4 had been applied. We found a significant effect of soil WFPS on 13C-CH4 oxidation rates and evidence for oxidation of 2.2 μg 13C-CH4 d−1 occurring in the 75% WFPS soil, which may have been either aerobic oxidation occurring in aerobic microsites in this soil or anaerobic CH4 oxidation. The lowest 13C-CH4 oxidation rate was measured in the 30% WFPS soil and was attributed to inhibition of methanotroph activity in this dry soil. However, oxidation was lowest in the wetter soils when estimated from changes in concentration of 12+13C-CH4. Thus, both methanogenesis and CH4 oxidation may have been occurring simultaneously in these wet soils, indicating the advantage of using a stable isotope approach to determine oxidation rates. Application of 13C-CH4 at 10 μl 13C-CH4 l−1 resulted in more rapid oxidation than under ambient CH4 conditions, suggesting CH4 oxidation in this soil was substrate limited, particularly in the wetter soils. Application of and (80 mg N kg soil−1; 9.9 at.% excess 15N) to different replicates enabled determination of the respective contributions of nitrification and denitrification to N2O emissions. The highest N2O emission (119 μg 14+15N-N2O kg soil−1 over 72 h) was measured from the 75% WFPS soil and was mostly produced during denitrification (18.1 μg 15N-N2O kg soil−1; 90% of 15N-N2O from this treatment). Strong negative correlations between 14+15N-N2O emissions, denitrified 15N-N2O emissions and 13C-CH4 concentrations (r=−0.93 to −0.95, N2O; r=−0.87 to −0.95, denitrified 15N-N2O; P<0.05) suggest a close relationship between CH4 oxidation and denitrification in our soil, the nature of which requires further investigation.  相似文献   

7.
We describe experiments to better understand how CH4 oxidation rates by different methanotroph communities respond to changing CH4 concentrations. We used a novel system of automatically monitored chambers to investigate the response of CH4 oxidation rates in a New Zealand pasture and adjacent pine forest soil exposed to varying atmospheric CH4 concentrations.Type II methanotrophs that dominate CH4 oxidation in the forest soil became progressively saturated as CH4 concentrations rose from ambient (1.8 ppmv) to 570 ppmv, as shown by a decrease in uptake efficiency from 20% to 2% removal. By contrast, CH4 oxidation in the pasture soil where Type I methanotrophs dominate increased in proportion to the increase in CH4 inlet concentration, oxidising about 2% of the inlet CH4 flux throughout. Modelling based on Michaelis-Menten kinetics revealed that low-affinity (Type I) methanotrophs were solely responsible for CH4 oxidation in pasture soils, whereas high affinity (Type II) methanotrophs only contributed about 10% of the CH4 oxidation in the forest soil. Increased aeration status using a soil–perlite (1:1) mixture doubled CH4 oxidation rates at both ambient (1.8 ppmv) and 40 ppmv atmospheric CH4. A similar volcanic soil previously exposed for 8 y to high CH4 fluxes from a landfill had removal efficiencies consistently above 95% for atmospheric CH4 concentrations up to 7500 ppmv when the CH4 oxidation rate was7000 μg CH4 kg−1soil h−1.  相似文献   

8.
Methane (CH4) is a critical greenhouse gas, with wetlands and flooded rice soils contributing >38% of the global emissions of CH4. A key process that offsets CH4 production is biological oxidation by methanotrophs that can consume as much as 90% of the CH4 produced in aerobic soils. The objective was to review the literature on the biochemical pathways, ecology of methanotrophs, research methods, and environmental and management controls on CH4 oxidation in wetlands. Methanotrophs have been extensively studied using phospholipid fatty acids (PLFA) but much less so using nucleic acid analysis or with the powerful stable-isotope probing technique of 13C-PLFA analysis. Of the environmental factors that affect CH4 oxidation, temperature, water content and redox potential are the most important whereas a wide range of soil pH enables oxidation. Methane oxidation varies with season as functions of temperature and plant community shifts (changes with growth stage of C input chemistry and oxygenation from aquatic rhizospheres). Indirect evidence suggests there is greater CH4 oxidation in pulsing than permanently flooded soils based on observations that static wetlands have greater emissions than pulsing wetlands. Basic research is needed on: the role of inorganic N species in controlling CH4 oxidation pathways; phylogenetic analysis of methanotrophs; and environmental and edaphic effects on methanotroph growth and activity. Research is needed to develop wetland systems that optimize CH4 oxidation relative to wetland type and environments and hydrology while maintaining and/or promoting other ecosystem services (e.g. C sequestration, pollution remediation, wildlife habitat, flood control).  相似文献   

9.

Purpose

Methane-oxidizing bacteria (methanotrophs) biologically consume and consequently affect the concentration of atmospheric methane (CH4), the second most prominent greenhouse gas, and therefore play critical roles in the mitigation of global warming effect. Long-term fertilization often affects the methanotrophic community and CH4 oxidation in various soils. Here, the immediate effects of nitrogen (N), phosphorus (P), and potassium (K) amendments on the CH4 oxidation activity and methanotrophic community structure were evaluated.

Materials and methods

Paddy soil samples were collected from the Taoyuan Experimental Station of the Chinese Academy of Sciences in central Hunan Province of China. A laboratory-based incubation experiment was conducted to investigate the immediate effects of N, P, and K amendments on the methanotrophs in soil. The CH4 oxidation rates and methanotrophic activities were determined by measuring the dynamic changes of CH4 concentration in the incubation system. The methanotrophic abundance and community changes in all of the seven treatments with and without nutrients addition were studied using real-time PCR and denaturing gradient gel electrophoresis, respectively.

Results and discussion

All of the N, P, and K treatments significantly decreased the CH4 oxidation activities. Compared with the control, the P and K amendments significantly increased the methanotrophic population size, but the N treatments have no effect on the methanotrophic abundance. A negative correlation was found between methanotrophic activity and methanotrophic abundance. We suggested that methanotrophic activity may not be inferred through the pmoA gene copies, especially in the short-term simulation experiments. Investigation of the methanotrophic population size and diversity is not enough to evaluate the soil CH4 sink accurately.

Conclusions

We concluded that the additions of N, P, and K reduce the activity but enhance the abundance of methanotrophs in a Chinese paddy soil through a short-term incubation experiment. Additionally, we found that the CH4 oxidation activity could be completely inhibited by Cl? toxicity. Our results implied that caution should be exercised in the types and amounts of fertilizers, especially KCl in agricultural systems to control the instantaneous increase in CH4 emission from the field.  相似文献   

10.
Agricultural factors affecting methane oxidation in arable soil   总被引：9，自引：0，他引：9  
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.  相似文献   

11.
Human activity has increased the amount of N entering terrestrial ecosystems from atmospheric NO3− deposition. High levels of inorganic N are known to suppress the expression of phenol oxidase, an important lignin-degrading enzyme produced by white-rot fungi. We hypothesized that chronic NO3− additions would decrease the flow of C through the heterotrophic soil food web by inhibiting phenol oxidase and the depolymerization of lignocellulose. This would likely reduce the availability of C from lignocellulose for metabolism by the microbial community. We tested this hypothesis in a mature northern hardwood forest in northern Michigan, which has received experimental atmospheric N deposition (30 kg NO3−-N ha−1 y−1) for nine years. In a laboratory study, we amended soils with 13C-labeled vanillin, a monophenolic product of lignin depolymerization, and 13C-labeled cellobiose, a disaccharide product of cellulose degradation. We then traced the flow of 13C through the microbial community and into soil organic carbon (SOC), dissolved organic carbon (DOC), and microbial respiration. We simultaneously measured the activity of enzymes responsible for lignin (phenol oxidase and peroxidase) and cellobiose (β-glucosidase) degradation. Nitrogen deposition reduced phenol oxidase activity by 83% and peroxidase activity by 74% when compared to control soils. In addition, soil C increased by 76%, whereas microbial biomass decreased by 68% in NO3− amended soils. 13C cellobiose in bacterial or fungal PLFAs was unaffected by NO3− deposition; however, the incorporation of 13C vanillin in fungal PLFAs extracted from NO3− amended soil was 82% higher than in the control treatment. The recovery of 13C vanillin and 13C cellobiose in SOC, DOC, microbial biomass, and respiration was not different between control and NO3− amended treatments. Chronic NO3− deposition has stemmed the flow of C through the heterotrophic soil food web by inhibiting the activity of ligninolytic enzymes, but it increased the assimilation of vanillin into fungal PLFAs.  相似文献   

12.
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.  相似文献   

13.
In a Japanese forest, CH4 uptake rate and methanotrophic community structure in the soil were investigated at four sites of different vegetation. At two of these sites, undergrowth was dominated by Sasa senanensis, and that of another was dominated by Sasa kurilensis. At the rest site, undergrowth had been removed artificially. The tree-layer composition differed between the two sites with S. senanensis, but tree layer of the other two sites were dominated by the same species. At the site lacking undergrowth, observed CH4 uptake rate was twice as high as that at the other sites. Under laboratory conditions, soil sample from the site lacking undergrowth exhibited CH4 consumption rate higher than that of the adjacent site with the same dominating tree species. The community structures of methanotrophs were investigated with denaturing gradient gel electrophoresis (DGGE) of the gene encoding particulate methane monooxygenase (pmoA). The banding patterns observed were different depending on the type of undergrowth vegetation. The sequences of the DGGE bands were closely related to each other and belonged to the “upland soil cluster alpha” (USCα). These results imply possible close relationship between the undergrowth vegetation and methanotrophic communities in forest soils.  相似文献   

14.
Methane oxidising activity and community structure of 11, specifically targeted, methanotrophic species have been examined in an arable soil. Soils were sampled from three different field plots, receiving no fertilisation (C), compost (G) and mineral fertiliser (M), respectively. Incubation experiments were carried out with and without pre-incubation at elevated CH4 mixing ratios (100 ml CH4 l−1) and with and without ammonium (100 mg N kg−1) pre-incubation. Four months after fertilisation, plots C, G and M did not show significant differences in physicochemical properties and CH4 oxidising activity. The total number of methanotrophs (determined as the sum the 11 specifically targeted methanotrophs) in the fresh soils was 17.0×106, 13.7×106 and 15.5×106 cells g−1 for treatment C, G and M, respectively. This corresponded to 0.11 to 0.32% of the total bacterial number. The CH4 oxidising activity increased 105-fold (20–26 mg CH4 g−1 h−1), the total number of methanotrophs doubled (28–76×106 cells g−1) and the methanotrophic diversity markedly increased in treatments with a pre-incubation at elevated CH4 concentrations. In all soils and treatments, type II methanotrophs (62–91%) outnumbered type I methanotrophs (9–38%). Methylocystis and Methylosinus species were always most abundant. After pre-incubation with ammonium, CH4 oxidation was completely inhibited; however, no change in the methanotrophic community structure could be detected.  相似文献   

15.
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.  相似文献   

16.
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.  相似文献   

17.
Methane production and consumption in a cultivated humisol   总被引：5，自引：0，他引：5  
Summary Laboratory studies were conducted on a cultivated humisol containing populations of both methanotrophs and methanogens. The molar ratio CO2 produced : O2 consumed :CH4 consumed was 0.27:1.0:1.0. Methane oxidation showed typical Michaelis-Menten kinetics with apparent K m values for CH4 and O2 of 66.2 M and 37.0 M, respectively. The low CO2 yields and the effects of low dissolved oxygen indicated the presence of aerobic obligate methanotrophs. It is suggested that the methanotrophs in this soil are not entirely dependent on atmospheric CH4 for growth and survival in situ.  相似文献   

18.
Root-derived rhizodeposits of recent photosynthetic carbon (C) are the foremost source of energy for microbial growth and development in rhizosphere soil. A substantial amount of photosynthesized C by the plants is translocated to belowground and is released as root exudates that influence the structure and function of soil microbial communities with potential inference in nutrient and C cycling in the ecosystem. We applied the 13C pulse chase labeling technique to evaluate the incorporation of rhizodeposit-C into the phospholipid fatty acids (PLFAs) in the bulk and rhizosphere soils of switchgrass (Panicum virgatum L.). Soil samples of bulk and rhizosphere were taken at 1, 5, 10 and 20 days after labeling and analyzed for 13C enrichment in the microbial PLFAs. Temporal differences of 13C enrichment in PLFAs were more prominent than spatial differences. Among the microbial PLFA biomarkers, fungi and Gram-negative (GM-ve) bacterial PLFAs showed rapid enrichment with 13C compared to Gram-positive (GM+ve) and actinomycetes in rhizosphere soil. The 13C enrichment of actinomycetes biomarker PLFA significantly increased along with sampling time in both soils. PLFAs indicative to fungi, GM-ve and GM+ve showed a significant decrease in 13C enrichment over sampling time in the rhizosphere, but a decrease was also observed in GM-ve (16:1ω5c) and fungal biomarker PLFAs in the bulk soil. The relative 13C concentration in fungal PLFA decreased on day 10, whereas those of GM-ve increased on day 5 and GM+ve remained constant in the rhizosphere soil. However, the relative 13C concentrations of GM-ve and GM+ve increased on days 5 and 10, respectively, and those of fungal remain constant in the bulk soil. The present study demonstrates the usefulness of 13C pulse chase labeling together with PLFA analysis to evaluate the active involvement of microbial community groups for utilizing rhizodeposit-C.  相似文献   

19.
A 13C natural abundance experiment including GC-c-IRMS analysis of phospholipid fatty acids (PLFAs) was conducted to assess the temporal dynamics of the soil microbial community and carbon incorporation during the mineralization of plant residues under the impact of heavy metals and acid rain. Maize straw was incorporated into (i) control soil, (ii) soil irrigated with acid rain, (iii) soil amended with heavy metal-polluted filter dust and (iv) soil with both, heavy metal and acid rain treatment, over a period of 74 weeks. The mineralization of maize straw carbon was significantly reduced by heavy metal impact. Reduced mineralization rate of the added carbon likely resulted from a reduction of the microbial biomass due to heavy metal stress, while the efficiency of 13C incorporation into microbial PLFAs was hardly affected. Since acid rain did not significantly change soil pH, little impact on soil microorganisms and mineralization rate was found. Temporal dynamics of labelling of microbial PLFAs were different between bacterial and fungal PLFA biomarkers. Utilization of maize straw by bacterial PLFAs peaked immediately after the application (2 weeks), while labelling of the fungal biomarker 18:2ω6,9 was most pronounced 5 weeks after the application. In general, 13C labelling of microbial PLFAs was closely linked to the amounts of maize carbon present in the soil. The distinct higher labelling of microbial PLFAs in the heavy metal-polluted soils 74 weeks after application indicated a large fraction of available maize straw carbon still present in the soil.  相似文献   

20.
Abstract

Methane flux was measured monthly from August 2002 to July 2003 at an oil palm plantation on tropical peatland in Sarawak, Malaysia, using a closed chamber technique. Urea was applied twice, once in November 2002 and once in May 2003. The monthly CH4 flux ranged from ?32.78 to 4.17 µg C m?2 h?1. Urea applications increased CH4 emissions in the month of application and emissions remained slightly higher a month later before the effect disappeared in the third month after application (i.e. back to CH4 uptake). This effect was the result of increased soil NH+ 4 content that was not immediately absorbed by the oil palm following urea application, which reduced the oxidation of CH4, resulting in its enhanced emission. By using the Cate–Nelson linear-plateau model, the critical soil NH+ 4 content causing CH4 emissions in the oil palm ecosystem was 42.75 mg kg?1 soil. However, the inhibitory effect of NH+ 4 on the oxidation of CH4 was mitigated by low rainfall and the pyrophosphate solubility index (PSI), where the former might increase oxidation of CH4 and the latter was a reflection of the low soluble substrate for methane production. Thus, the splitting and timing of urea applications are important not only to optimize oil palm yield, but also to reduce soil NH+ 4 content to minimize CH4 emissions and, therefore, its potential negative impact on the environment.  相似文献   

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