Methanotrophs,organisms that obtain oxygen by oxidizing methane,are recognized as the only known biological sink for atmospheric CH_4,and forest soil methanotrophs play crucial roles in mitigating global warming.The s...Methanotrophs,organisms that obtain oxygen by oxidizing methane,are recognized as the only known biological sink for atmospheric CH_4,and forest soil methanotrophs play crucial roles in mitigating global warming.The succession patterns of methanotrophic communities and functions in Wudalianchi volcano forest soils could provide a basis for the study of evolutionary mechanisms between soil microorganisms,the environment,and carbon cycling of temperate forest ecosystems under climate change.In this study,the characteristics and drivers of methanotrophic community structure and function of two volcanic soils at different stages of development are analyzed,including an old volcano and a new volcano,which most recently erupted 300 years and 17-19×10^(5)years ago,respectively,and a non-volcano hills as control,based on space for time substitution and Miseq sequencing and bioinformation technology.The results showed that CH_(4) fluxes were significantly higher in old-stage volcano forest soils than new-stage forest soils and non-volcano forest soils.There were significant differences in the community composition and diversity of soil methanotrophs from different volcano forest soils.Methylococcus was the dominant genus in all soil samples.Additionally,the relative abundance of Methylococcus,along with Clonothrix,Methyloglobulus,Methylomagum,Methylomonas and Methylosarcina,were the important genera responsible for the differences in methanotrophic community structure in different volcano forest soils.The relative abundance of methanotroph belonging toγ-proteobacteria was significantly higher than that belonging toα-proteobacteria(P<0.05).Chao1,Shannon and Simpson indices of soil methanotrophic community were significantly lower in new-stage volcanos and were significantly affected by bulk density,total porosity,p H,nitrate,dissolved organic carbon and dissolved organic nitrogen.There were significant differences in community structure between new-stage and old-stage volcanoes.Bulk density and p H are important soil properties contributing to the divergence of methanotrophs community structure,and changes in soil properties due to soil development time are important factors driving differences in methanotrophs communities in Wudalianchi volcanic soils.展开更多
Poly-3-hydroxybutyrate (PHB) can be produced by various species of bacteria. Among the possible carbon sources, both methane and methanol could be a suitable substrate for the production of PHB. Methane is cheap and...Poly-3-hydroxybutyrate (PHB) can be produced by various species of bacteria. Among the possible carbon sources, both methane and methanol could be a suitable substrate for the production of PHB. Methane is cheap and plentiful not only as natural gas, but also as biogas. Methanol can also maintain methanotrophic activity in some conditions. The methanotrophic strain Methylosinus trichosporium IMV3011 can accumulate PHB with methane and methanol in a brief nonsterile process. Liquid methanol (0.1%) was added to improve the oxidization of methane. The studies were carried out using shake flasks. Cultivation was performed in two stages: a continuous growth phase and a PHB accumulation phase under the conditions short of essential nutrients (ammonium, nitrate, phosphorus, copper, iron (Ⅲ), magnesium or ethylenediamine tetraacetate (EDTA)) in batch culture. It was found that the most suitable growth time for the cell is 144 h. Then an optimized culture condition for second stage was determined, in which the PHB concentration could be much increased to 0.6 g/L. In order to increase PHB content, citric acid was added as an inhibitor of tricarboxylic acid cycle (TCA). It was found that citric acid is favorable for the PHB accumulation, and the PHB yield was increased to 40% (w/w) from the initial yield of 12% (w/w) after nutrient deficiency cultivation. The PHB produced is of very high quality with molecular weight up to 1.5 × 10^6Da.展开更多
Methanotrophs in marine sediments and overlying water attenuate the emissions of methane into the atmosphere and thus play an important role for the global cycle of this greenhouse gas.However,gas released from natura...Methanotrophs in marine sediments and overlying water attenuate the emissions of methane into the atmosphere and thus play an important role for the global cycle of this greenhouse gas.However,gas released from natural hydrocarbon seeps are not pure methane but commonly mixed hydrocarbons.Currently,how methanotrophic bacteria behave in the co-presence of methane and heavier hydrocarbons remains unknown.In this paper,the bacteria were cultured aerobically in fresh sediment samples(collected from Bohai Bay in eastern China)at 28℃under the atmospheres of pure methane and methane+ethane+propane mixed gas,respec-tively.The prevailing terrigenous n-alkanes and fatty acids in the original sediment samples varied consistently after incubations,confirming the proceeding of aerobic bacterial activities.The real-time quantitative PCR assay and sequencing of the 16S rRNA and particulate methane monooxygenase(pmoA)genes revealed the changes of microbe communities to a methanotroph-dominating structure after incubations.Particularly,after incubations the family Methylococcaceae(typeⅠmethanotrophs)became dominant with proportions higher than 40%,whereas Methylocystaceae(typeⅡmethanotrophs)nearly disappeared in all incubated samples.More-over,the species of methanotrophs from the samples treated with pure methane were dominated by Methylobacter luteus,whereas Methylobacter whittenburyi took the predominant proportion in the samples treated with mixed gas.The phenomenon suggests that some methanotrophs may also utilize ethane and propane.Collectively,this study may help to gain a better understanding of the ef-fects and contributions of microbial activities in marine hydrocarbon seep ecosystems.展开更多
Wetland ecosystems are the most important natural methane(CH_(4))sources,whose fluxes periodically fluctuate.Methanogens(methane producers)and methanotrophs(methane consumers)are considered key factors affecting CH_(4...Wetland ecosystems are the most important natural methane(CH_(4))sources,whose fluxes periodically fluctuate.Methanogens(methane producers)and methanotrophs(methane consumers)are considered key factors affecting CH_(4)fluxes in wetlands.However,the symbiotic relationship between methanogens and methanotrophs remains unclear.To help close this research gap,we collected and analyzed samples from four soil depths in the Dajiuhu subalpine peatland in January,April,July,and October 2019 and acquired seasonal methane flux data from an eddy covariance(EC)system,and investigated relationships.A phylogenetic molecular ecological networks(pMENs)analysis was used to identify keystone species and the seasonal variations of the co-occurrence patterns of methanogenic and methanotrophic communities.The results indicate that the seasonal variations of the interactions between methanogenic and methanotrophic communities contributed to CH_(4)emissions in wetlands.The keystone species discerned by the network analysis also showed their importance in mediating CH_(4)fluxes.Methane(CH_(4))emissions in wetlands were lowest in spring;during this period,the most complex interactions between microbes were observed,with intense competition among methanogens while methanotrophs demonstrated better cooperation.Reverse patterns manifested themselves in summer when the highest CH_(4)flux was observed.Methanoregula formicica was negatively correlated with CH_(4)fluxes and occupied the largest ecological niches in the spring network.In contrast,both Methanocella arvoryzae and Methylocystaceae demonstrated positive correlations with CH_(4)fluxes and were better adapted to the microbial community in the summer.In addition,soil temperature and nitrogen were regarded as significant environmental factors to CH_(4)fluxes.This study was successful in explaining the seasonal patterns and microbial driving mechanisms of CH_(4)emissions in wetlands.展开更多
Methanotrophs play a vital role in the mitigation of methane emission from soils. However, the influences of cover crops incorporation on paddy soil methanotrophic community structure have not been fully understood. I...Methanotrophs play a vital role in the mitigation of methane emission from soils. However, the influences of cover crops incorporation on paddy soil methanotrophic community structure have not been fully understood. In this study, the impacts of two winter cover crops(Chinese milk vetch(Astragalus sinicus L.) and ryegrass(Lolium multiflorum Lam.), representing leguminous and non-leguminous cover crops, respectively) on community structure and abundance of methanotrophs were evaluated by using PCR-DGGE(polymerase chain reaction-denaturing gradient gel electrophoresis) and real-time PCR technology in a double-rice cropping system from South China. Four treatments were established in a completely randomized block design: 1) double-rice cropping without nitrogen fertilizer application, CK; 2) double-rice cropping with chemical nitrogen fertilizer application(200 kg ha^(–1) urea for entire double-rice season), CF; 3) Chinese milk vetch cropping followed by double-rice cultivation with Chinese milk vetch incorporation, MV; 4) ryegrass cropping followed by double-rice cultivation with ryegrass incorporation, RG. Results showed that cultivating Chinese milk vetch and ryegrass in fallow season decreased soil bulk density and increased rice yield in different extents by comparison with CK. Additionally, methanotrophic bacterial abundance and community structure changed significantly with rice growth. Methanotrophic bacterial pmo A gene copies in four treatments were higher during late-rice season(3.18×10^7 to 10.28×10^7 copies g^–1 dry soil) by comparison with early-rice season(2.1×10^7 to 9.62×10^7 copies g^–1 dry soil). Type Ⅰ methanotrophs absolutely predominated during early-rice season. However, the advantage of type Ⅰ methanotrophs kept narrowing during entire double-rice season and both types Ⅰ and Ⅱ methanotrophs dominated at later stage of late-rice.展开更多
Large amounts of gas hydrate are distributed in the northern slope of the South China Sea,which is a potential threat of methane leakage.Aerobic methane oxidation by methanotrophs,significant methane biotransformation...Large amounts of gas hydrate are distributed in the northern slope of the South China Sea,which is a potential threat of methane leakage.Aerobic methane oxidation by methanotrophs,significant methane biotransformation that occurs in sediment surface and water column,can effectively reduce atmospheric emission of hydrate-decomposed methane.To identify active aerobic methanotrophs and their methane oxidation potential in sediments from the Shenhu Area in the South China Sea,multi-day enrichment incubations were conducted in this study.The results show that the methane oxidation rates in the studied sediments were 2.03‒2.36μmol/gdw/d,which were higher than those obtained by sediment incubations from other areas in marine ecosystems.Thus the authors suspect that the methane oxidation potential of methanotrophs was relatively higher in sediments from the Shenhu Area.After the incubations family Methylococcaea(type I methanotrophs)mainly consisted of genus Methylobacter and Methylococcaea_Other were predominant with an increased proportion of 70.3%,whereas Methylocaldum decreased simultaneously in the incubated sediments.Collectively,this study may help to gain a better understanding of the methane biotransformation in the Shenhu Area.展开更多
Background:Methane(CH4)oxidation driven by soil aerobic methanotrophs demonstrates the capacity of grassland as a CH4 sink.Methods:In this study,we compared the oxidation characteristics of atmospheric-level and eleva...Background:Methane(CH4)oxidation driven by soil aerobic methanotrophs demonstrates the capacity of grassland as a CH4 sink.Methods:In this study,we compared the oxidation characteristics of atmospheric-level and elevated concentration(10%)CH4 in a typical grassland(steppe)on the Loess Plateau,an alpine meadow(meadow)on the Qinghai-Tibet Plateau,and an inland arid-area artificial grassland(pasture)in northwest China and investigated the communities of active methanotrophs and their contribution to CH4 oxidation using DNA-based stable-isotope probing and Illumina Miseq sequencing.Results:The results showed that the oxidation of atmospheric CH4 only occurred in steppe and meadow soils where the USCγgroup of methanotrophs was numerically dominant in the methanotroph community.Pasture soils,with their very low relative abundance of USCγ,did not show atmospheric CH4 oxidation.However,a DNA-stable isotope probing experiment with 10%CH4 indicated that conventional CH4 oxidizers(Methylocaldum and Methylocystis)rather than USCγcommunities assimilated significant amounts of 13CH4 for growth.Conclusions:The CH4 oxidation mechanisms in the three experimental grassland soils varied significantly.The USCγgroup may be obligate oligotrophic microorganisms or their growth requires specific unknown conditions.展开更多
Methane(CH_(4))is a potent greenhouse gas that has a substantial impact on global warming due to its substantial influence on the greenhouse effect.Increasing extreme precipitation events,such as drought,attributable ...Methane(CH_(4))is a potent greenhouse gas that has a substantial impact on global warming due to its substantial influence on the greenhouse effect.Increasing extreme precipitation events,such as drought,attributable to global warming that caused by greenhouse gases,exert a profound impact on the intricate biological processes associated with CH_(4) uptake.Notably,the timing of extreme drought occurrence emerges as a pivotal factor influencing CH_(4) uptake,even when the degree of drought remains constant.However,it is still unclear how the growing season regulates the response of CH_(4) uptake to extreme drought.In an effort to bridge this knowledge gap,we conducted a field manipulative experiment to evaluate the impact of extreme drought on CH_(4) uptake during early,middle,and late growing stages in a temperate steppe of Inner Mongolia Autonomous Region,China.The result showed that all extreme drought consistently exerted positive effects on CH_(4) uptake regardless of seasonal timing.However,the magnitude of this effect varied depending on the timing of season,as evidenced by a stronger effect in early growing stage than in middle and late growing stages.Besides,the pathways of CH_(4) uptake were different from seasonal timing.Extreme drought affected soil physical-chemical properties and aboveground biomass(AGB),consequently leading to changes in CH_(4) uptake.The structural equation model showed that drought both in the early and middle growing stages enhanced CH_(4) uptake due to reduced soil water content(SWC),leading to a decrease in NO_(3)–-N and an increase in pmoA abundance.However,drought in late growing stage primarily enhanced CH_(4) uptake only by decreasing SWC.Our results suggested that seasonal timing significantly contributed to regulate the impacts of extreme drought pathways and magnitudes on CH_(4) uptake.The findings can provide substantial implications for understanding how extreme droughts affect CH_(4) uptake and improve the prediction of potential ecological consequence under future climate change.展开更多
Zoige wetland is one of the most important methane emission centers in China. The oxidation of methane in the wetland affects global warming, soil ecology and atmospheric chemistry. Despite their global significance, ...Zoige wetland is one of the most important methane emission centers in China. The oxidation of methane in the wetland affects global warming, soil ecology and atmospheric chemistry. Despite their global significance, microorganisms that consume methane in Zoige wetland remain poorly characterized. In this study, we investigated methanotrophs diversity in soil samples from both anaerobic site and aerobic site in Zoige wetland using pmoA gene as a molecular marker. The cloning library was constructed according to the pmoA sequences detected. Four clusters of methanotrophs were detected. The phylogenetic tree showed that all four clusters detected were affiliated to type I methanotrophs. Two novel clusters (cluster 1, cluster 2) were found to relate to none of the recognized genera of methanotrophs. These clusters have no cultured representatives and reveal an ecological adaptation of particular uncultured methanotrophs in Zoige wetland. Two clusters were belonging to Methylobacter and Methylococcus separately. Denaturing gradient gel electrophoresis gel bands pattern retrieved from these two samples revealed that the community compositions of anaerobic soil and aerobic soil were different from each other while anaerobic soil showed a higher metanotrophs diversity. Real-time PCR assays of the two samples demonstrated that aerobic soil sample in Zoige wetland was 1.5 times as much copy numbers as anaerobic soil. These data illustrated that methanotrophs are a group of microorganisms influence the methane consumption in Zoige wetland.展开更多
Global warming, as a result of an increase in the mean temperature of the planet, might lead to catastrophic events for humanity. This temperature increase is mainly the result of an increase in the atmospheric greenh...Global warming, as a result of an increase in the mean temperature of the planet, might lead to catastrophic events for humanity. This temperature increase is mainly the result of an increase in the atmospheric greenhouse gases (GHG) concentration. Water vapor, carbon dioxide (CO2), methane (CH4) and nitrous oxide (N20) are the most important GHG, and human activities, such as industry, livestock and agriculture, contribute to the production of these gases. Methane, at an atmospheric concentration of 1.7 gmol tool-1 currently, is responsible for 16% of the global warming due to its relatively high global warming potential. Soils play an important role in the CH4 cycle as methanotrophy (oxidation of CH4) and methanogenesis (production of CH4) take place in them. Understanding methanogenesis and methanotrophy is essential to establish new agriculture techniques and industrial processes that contribute to a better balance of GHG. The current knowledge of methanogenesis and methanotrophy in soils, anaerobic CH4 oxidation and methanotrophy in extreme environments is also discussed.展开更多
Soil samples were collected from three plots under different land utilization patterns including degradation, farming, and restoration. The abundances of methanotrophs were quantified using real-time polymerase chain ...Soil samples were collected from three plots under different land utilization patterns including degradation, farming, and restoration. The abundances of methanotrophs were quantified using real-time polymerase chain reaction (PCR) based on the pmoA and 16S rRNA genes, and the community fingerprint was analyzed using denaturing gradient gel electrophoresis (DGGE) aiming at pmoA gene. Significantly lower 16S rRNA and pmoA genes copies were found in the degradation treatment than in farming and restoration. Higher abundances of Type I than those of Type II methanotrophs were detected in all treatments, The treatment of farming was clearly separated from degradation and restoration according to the DGGE profile by cluster analysis. The lowest diversity indices were observed in the F (farming plot), suggesting that the community structure was strongly affected by farming activities. There were significantly positive correlations between the copy numbers of pmoA also Type II-related 16S rRNA genes and soil available K content. Strong negative and positive correlations were found between Type I and soil pH, and available P content, respectively. We concluded that the vegetation cover or not, soil characteristics including pH and nutrients of P and K as a result of anthropogenic disturbance may be key factors affecting methanotrophic communities in upland soil.展开更多
Aged refuse from waste landfills closed for eight years was examined and found to contain rich methanotrophs capable of biooxidation for methane. Specially, community structure and methane oxidation capability of meth...Aged refuse from waste landfills closed for eight years was examined and found to contain rich methanotrophs capable of biooxidation for methane. Specially, community structure and methane oxidation capability of methanotrophs in the aged refuse were studied. The amount of methanotrophs ranged 61.97×10^3-632.91×10^3 cells/g (in dry basis) in aged refuse from Shanghai Laogang Landfill. Type I and II methanotrophs were found in the aged refuse in the presence of sterilized sewage sludge and only Type I methanotrophs were detected in the presence of nitrate minimal salt medium (NMS). The clone sequences of the pmoA gene obtained from the aged refuse were similar to the pmoA gene of Methylobacter, Methylocaldum, and Methylocystis, and two clones were distinct with known genera of Type I methanotrophs according to phylogenetic analysis. Aged refuse enriched with NMS was used for methane biological oxidation and over 93% conversions were obtained.展开更多
Methanotrophic–heterotrophic communities were selectively enriched from sewage sludge to obtain a mixed culture with high levels of poly-β-hydroxybutyrate(PHB)accumulation capacity from methane.Methane was used as...Methanotrophic–heterotrophic communities were selectively enriched from sewage sludge to obtain a mixed culture with high levels of poly-β-hydroxybutyrate(PHB)accumulation capacity from methane.Methane was used as the carbon source,N2as sole nitrogen source,and oxygen and Cu content were varied.Copper proved essential for PHB synthesis.All cultures enriched with Cu could accumulate high content of PHB(43.2%–45.9%),while only small amounts of PHB were accumulated by cultures enriched without Cu(11.9%–17.5%).Batch assays revealed that communities grown with Cu and a higher O2content synthesized more PHB,which had a wider optimal CH4:O2range and produced a high PHB content(48.7%)even though in the presence of N2.In all methanotrophic–heterotrophic communities,both methanotrophic and heterotrophic populations showed the ability to accumulate PHB.Although methane was added as the sole carbon source,heterotrophs dominated with abundances between 77.2%and 85.6%.All methanotrophs detected belonged to type II genera,which formed stable communities with heterotrophs of different PHB production capacities.展开更多
Methanotrophic bacteria are entities with innate biocatalytic potential to biofilter and oxidize methane into simpler compounds concomitantly conserving energy,which can contribute to copious industrial applications.T...Methanotrophic bacteria are entities with innate biocatalytic potential to biofilter and oxidize methane into simpler compounds concomitantly conserving energy,which can contribute to copious industrial applications.The future and efficacy of such industrial applications relies upon acquiring and/or securing robust methanotrophs with taxonomic and phenotypic diversity.Despite several dramatic advances,isolation of robust methanotrophs is still a long-way challenging task with several lacunae to be filled in sequentially.Methanotrophs with high tolerance to methane can be isolated and cultivated by mimicking natural environs,and adopting strategies like adaptive metabolic evolution.This review summarizes existent and innovative methods for methanotrophic isolation and purification,and their respective applications.A comprehensive description of new insights shedding light upon how to isolate and concomitantly augment robust methanotrophic metabolism in an orchestrated fashion follows.展开更多
Methanotrophic based process can be the remedy to offset the wastewater treatment facilities increasing energy requirements due to methanotroph's unique ability to integrate methane assimilation with multiple biotech...Methanotrophic based process can be the remedy to offset the wastewater treatment facilities increasing energy requirements due to methanotroph's unique ability to integrate methane assimilation with multiple biotechnological applications like biological nitrogen removal and methanol production. Regardless of the methanotrophic process end product, the challenge to maintain stable microbial growth in the methanotrophs cultivation bioreactor at higher cell densities is one of the major obstacles facing the process upscaling. Therefore, a series of consecutive batch tests were performed to attentively investigate the biomass density influence on type I methanotrophs bacterial growth. In addition, food to microorganisms(F/M), carbon to nitrogen(C/N) and nitrogen to microorganisms(N/M) ratio effect on the microbial activity was studied for the first time. It was clarified that the F/M ratio is the most influencing factor on the microbial growth at higher biomass densities rather than the biomass density increase, whereas C/N and N/M ratio change, while using nitrate as the nitrogen source,does not influence methanotrophs microbial growth. These study results would facilitate the scaling up of methanotrophic based biotechnology by identifying that F/M ratio as the key parameter that influences methanotrophs cultivation at high biomass densities.展开更多
Biological methane oxidation is a crucial process in the global carbon cycle that reduces methane emissions from paddy fields and natural wetlands into the atmosphere.However,soil organic carbon accumulation associate...Biological methane oxidation is a crucial process in the global carbon cycle that reduces methane emissions from paddy fields and natural wetlands into the atmosphere.However,soil organic carbon accumulation associated with microbial methane oxidation is poorly understood.Therefore,to investigate methane-derived carbon incorporation into soil organic matter,paddy soils originated from different parent materials(Inceptisol,Entisol,and Alfisol) were collected after rice harvesting from four major rice-producing regions in Bangladesh.Following microcosm incubation with 5%(volume/volume)^(13) CH_(4),soil^(13) C-atom abundances significantly increased from background level of 1.08% to 1.88%–2.78%,leading to a net methane-derived accumulation of soil organic carbon ranging from 120 to 307 mg kg^(-1).Approximately 23.6%–60.0% of the methane consumed was converted to soil organic carbon during microbial methane oxidation.The phylogeny of^(13) C-labeled pmoA(enconding the alpha subunit of the particulate methane monooxygenase) and 16 S rRNA genes further revealed that canonical α(type II) and γ(type I) Proteobacteria were active methane oxidizers.Members within the Methylobacter-and Methylosarcina-affiliated type Ia lineages dominated active methane-oxidizing communities that were responsible for the majority of methane-derived carbon accumulation in all three paddy soils,while Methylocystis-affiliated type IIa lineage was the key contributor in one paddy soil of Inceptisol origin.These results suggest that methanotroph-mediated synthesis of biomass plays an important role in soil organic matter accumulation.This study thus supports the concept that methanotrophs not only consume the greenhouse gas methane but also serve as a key biotic factor in maintaining soil fertility.展开更多
A simulated landfill biocover microcosm consisting of a modifying ceramsite material and compost were investigated.Results show that the mixture can improve the material porosity and achieve a stable and highly effici...A simulated landfill biocover microcosm consisting of a modifying ceramsite material and compost were investigated.Results show that the mixture can improve the material porosity and achieve a stable and highly efficient (100%) methane oxidation over an extended operating period.The diversity of the methanotrophic community in the microcosm was assessed.Type I methanotrophs were enhanced in the microcosm due to the increased air diffusion and distribution,whereas the microbial diversity and population density of type II methanotrophs were not significantly affected.Moreover,the type I methanotrophic community structure significantly varied with the reactor height,whereas that of type II methanotrophic communities did not exhibit a spatial variation.Phylogenetic analysis showed that type I methanotroph-based nested polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) resulted in the detection of eight different populations,most of which are related to Methylobacter sp.,whereas that of type II resulted in the detection of nine different populations,most of which are related to Methylocystaceae.Methanotrophic community analysis also indicated that a number of new methanotrophic genera not closely related to any known methanotrophic populations were present.展开更多
基金the Special Projects for the Central Government to guide the development of local science and technology(ZY20B15)the Key Research&Development Program funding project of Heilongjiang Province(GA21C030)the Research Funds of Provincial Research Institutes of Heilongjiang Province(ZNBZ2022ZR07)。
文摘Methanotrophs,organisms that obtain oxygen by oxidizing methane,are recognized as the only known biological sink for atmospheric CH_4,and forest soil methanotrophs play crucial roles in mitigating global warming.The succession patterns of methanotrophic communities and functions in Wudalianchi volcano forest soils could provide a basis for the study of evolutionary mechanisms between soil microorganisms,the environment,and carbon cycling of temperate forest ecosystems under climate change.In this study,the characteristics and drivers of methanotrophic community structure and function of two volcanic soils at different stages of development are analyzed,including an old volcano and a new volcano,which most recently erupted 300 years and 17-19×10^(5)years ago,respectively,and a non-volcano hills as control,based on space for time substitution and Miseq sequencing and bioinformation technology.The results showed that CH_(4) fluxes were significantly higher in old-stage volcano forest soils than new-stage forest soils and non-volcano forest soils.There were significant differences in the community composition and diversity of soil methanotrophs from different volcano forest soils.Methylococcus was the dominant genus in all soil samples.Additionally,the relative abundance of Methylococcus,along with Clonothrix,Methyloglobulus,Methylomagum,Methylomonas and Methylosarcina,were the important genera responsible for the differences in methanotrophic community structure in different volcano forest soils.The relative abundance of methanotroph belonging toγ-proteobacteria was significantly higher than that belonging toα-proteobacteria(P<0.05).Chao1,Shannon and Simpson indices of soil methanotrophic community were significantly lower in new-stage volcanos and were significantly affected by bulk density,total porosity,p H,nitrate,dissolved organic carbon and dissolved organic nitrogen.There were significant differences in community structure between new-stage and old-stage volcanoes.Bulk density and p H are important soil properties contributing to the divergence of methanotrophs community structure,and changes in soil properties due to soil development time are important factors driving differences in methanotrophs communities in Wudalianchi volcanic soils.
基金New Century Excellent Talents in University of China(NCET-05-0358)the National Natural Science Foundation of China(20625308)
文摘Poly-3-hydroxybutyrate (PHB) can be produced by various species of bacteria. Among the possible carbon sources, both methane and methanol could be a suitable substrate for the production of PHB. Methane is cheap and plentiful not only as natural gas, but also as biogas. Methanol can also maintain methanotrophic activity in some conditions. The methanotrophic strain Methylosinus trichosporium IMV3011 can accumulate PHB with methane and methanol in a brief nonsterile process. Liquid methanol (0.1%) was added to improve the oxidization of methane. The studies were carried out using shake flasks. Cultivation was performed in two stages: a continuous growth phase and a PHB accumulation phase under the conditions short of essential nutrients (ammonium, nitrate, phosphorus, copper, iron (Ⅲ), magnesium or ethylenediamine tetraacetate (EDTA)) in batch culture. It was found that the most suitable growth time for the cell is 144 h. Then an optimized culture condition for second stage was determined, in which the PHB concentration could be much increased to 0.6 g/L. In order to increase PHB content, citric acid was added as an inhibitor of tricarboxylic acid cycle (TCA). It was found that citric acid is favorable for the PHB accumulation, and the PHB yield was increased to 40% (w/w) from the initial yield of 12% (w/w) after nutrient deficiency cultivation. The PHB produced is of very high quality with molecular weight up to 1.5 × 10^6Da.
基金This work was supported by the Natural Science Foun-dation of Shandong Province(No.ZR2020QD070)the National Natural Science Foundation of China(No.41876051)the China Geological Survey Project(No.DD 20190221).
文摘Methanotrophs in marine sediments and overlying water attenuate the emissions of methane into the atmosphere and thus play an important role for the global cycle of this greenhouse gas.However,gas released from natural hydrocarbon seeps are not pure methane but commonly mixed hydrocarbons.Currently,how methanotrophic bacteria behave in the co-presence of methane and heavier hydrocarbons remains unknown.In this paper,the bacteria were cultured aerobically in fresh sediment samples(collected from Bohai Bay in eastern China)at 28℃under the atmospheres of pure methane and methane+ethane+propane mixed gas,respec-tively.The prevailing terrigenous n-alkanes and fatty acids in the original sediment samples varied consistently after incubations,confirming the proceeding of aerobic bacterial activities.The real-time quantitative PCR assay and sequencing of the 16S rRNA and particulate methane monooxygenase(pmoA)genes revealed the changes of microbe communities to a methanotroph-dominating structure after incubations.Particularly,after incubations the family Methylococcaceae(typeⅠmethanotrophs)became dominant with proportions higher than 40%,whereas Methylocystaceae(typeⅡmethanotrophs)nearly disappeared in all incubated samples.More-over,the species of methanotrophs from the samples treated with pure methane were dominated by Methylobacter luteus,whereas Methylobacter whittenburyi took the predominant proportion in the samples treated with mixed gas.The phenomenon suggests that some methanotrophs may also utilize ethane and propane.Collectively,this study may help to gain a better understanding of the ef-fects and contributions of microbial activities in marine hydrocarbon seep ecosystems.
基金supported by the National Science Foundation of China(Grant No.31971490).
文摘Wetland ecosystems are the most important natural methane(CH_(4))sources,whose fluxes periodically fluctuate.Methanogens(methane producers)and methanotrophs(methane consumers)are considered key factors affecting CH_(4)fluxes in wetlands.However,the symbiotic relationship between methanogens and methanotrophs remains unclear.To help close this research gap,we collected and analyzed samples from four soil depths in the Dajiuhu subalpine peatland in January,April,July,and October 2019 and acquired seasonal methane flux data from an eddy covariance(EC)system,and investigated relationships.A phylogenetic molecular ecological networks(pMENs)analysis was used to identify keystone species and the seasonal variations of the co-occurrence patterns of methanogenic and methanotrophic communities.The results indicate that the seasonal variations of the interactions between methanogenic and methanotrophic communities contributed to CH_(4)emissions in wetlands.The keystone species discerned by the network analysis also showed their importance in mediating CH_(4)fluxes.Methane(CH_(4))emissions in wetlands were lowest in spring;during this period,the most complex interactions between microbes were observed,with intense competition among methanogens while methanotrophs demonstrated better cooperation.Reverse patterns manifested themselves in summer when the highest CH_(4)flux was observed.Methanoregula formicica was negatively correlated with CH_(4)fluxes and occupied the largest ecological niches in the spring network.In contrast,both Methanocella arvoryzae and Methylocystaceae demonstrated positive correlations with CH_(4)fluxes and were better adapted to the microbial community in the summer.In addition,soil temperature and nitrogen were regarded as significant environmental factors to CH_(4)fluxes.This study was successful in explaining the seasonal patterns and microbial driving mechanisms of CH_(4)emissions in wetlands.
基金supported by the National Natural Science Foundation of China (31171509 and 30671222)the Special Fund for Agro-scientific Research in the Public Interest, China (201103001)the National Key Technology R&D Program for the 12th Five-Year Plan period (2011BAD16B15)
文摘Methanotrophs play a vital role in the mitigation of methane emission from soils. However, the influences of cover crops incorporation on paddy soil methanotrophic community structure have not been fully understood. In this study, the impacts of two winter cover crops(Chinese milk vetch(Astragalus sinicus L.) and ryegrass(Lolium multiflorum Lam.), representing leguminous and non-leguminous cover crops, respectively) on community structure and abundance of methanotrophs were evaluated by using PCR-DGGE(polymerase chain reaction-denaturing gradient gel electrophoresis) and real-time PCR technology in a double-rice cropping system from South China. Four treatments were established in a completely randomized block design: 1) double-rice cropping without nitrogen fertilizer application, CK; 2) double-rice cropping with chemical nitrogen fertilizer application(200 kg ha^(–1) urea for entire double-rice season), CF; 3) Chinese milk vetch cropping followed by double-rice cultivation with Chinese milk vetch incorporation, MV; 4) ryegrass cropping followed by double-rice cultivation with ryegrass incorporation, RG. Results showed that cultivating Chinese milk vetch and ryegrass in fallow season decreased soil bulk density and increased rice yield in different extents by comparison with CK. Additionally, methanotrophic bacterial abundance and community structure changed significantly with rice growth. Methanotrophic bacterial pmo A gene copies in four treatments were higher during late-rice season(3.18×10^7 to 10.28×10^7 copies g^–1 dry soil) by comparison with early-rice season(2.1×10^7 to 9.62×10^7 copies g^–1 dry soil). Type Ⅰ methanotrophs absolutely predominated during early-rice season. However, the advantage of type Ⅰ methanotrophs kept narrowing during entire double-rice season and both types Ⅰ and Ⅱ methanotrophs dominated at later stage of late-rice.
基金jointly supported by the National Natural Science Foundation of China (42106052)Shandong Provincial Natural Science Foundation (ZR2020QD070)the project of China Geological Survey (DD20190221)。
文摘Large amounts of gas hydrate are distributed in the northern slope of the South China Sea,which is a potential threat of methane leakage.Aerobic methane oxidation by methanotrophs,significant methane biotransformation that occurs in sediment surface and water column,can effectively reduce atmospheric emission of hydrate-decomposed methane.To identify active aerobic methanotrophs and their methane oxidation potential in sediments from the Shenhu Area in the South China Sea,multi-day enrichment incubations were conducted in this study.The results show that the methane oxidation rates in the studied sediments were 2.03‒2.36μmol/gdw/d,which were higher than those obtained by sediment incubations from other areas in marine ecosystems.Thus the authors suspect that the methane oxidation potential of methanotrophs was relatively higher in sediments from the Shenhu Area.After the incubations family Methylococcaea(type I methanotrophs)mainly consisted of genus Methylobacter and Methylococcaea_Other were predominant with an increased proportion of 70.3%,whereas Methylocaldum decreased simultaneously in the incubated sediments.Collectively,this study may help to gain a better understanding of the methane biotransformation in the Shenhu Area.
基金National Natural Science Foundation of China,Grant/Award Numbers:42277114,91751204,41877062。
文摘Background:Methane(CH4)oxidation driven by soil aerobic methanotrophs demonstrates the capacity of grassland as a CH4 sink.Methods:In this study,we compared the oxidation characteristics of atmospheric-level and elevated concentration(10%)CH4 in a typical grassland(steppe)on the Loess Plateau,an alpine meadow(meadow)on the Qinghai-Tibet Plateau,and an inland arid-area artificial grassland(pasture)in northwest China and investigated the communities of active methanotrophs and their contribution to CH4 oxidation using DNA-based stable-isotope probing and Illumina Miseq sequencing.Results:The results showed that the oxidation of atmospheric CH4 only occurred in steppe and meadow soils where the USCγgroup of methanotrophs was numerically dominant in the methanotroph community.Pasture soils,with their very low relative abundance of USCγ,did not show atmospheric CH4 oxidation.However,a DNA-stable isotope probing experiment with 10%CH4 indicated that conventional CH4 oxidizers(Methylocaldum and Methylocystis)rather than USCγcommunities assimilated significant amounts of 13CH4 for growth.Conclusions:The CH4 oxidation mechanisms in the three experimental grassland soils varied significantly.The USCγgroup may be obligate oligotrophic microorganisms or their growth requires specific unknown conditions.
基金This study was funded by the National Natural Science Foundation of China(42041005,U20A2050,U21A20240)the Weiqiao-UCAS(University of Chinese Academy of Sciences)Special Projects on Low-Carbon Technology Development(GYY-DTFZ-2022-006)the Fundamental Research Funds for the Central Universities(E1E40607).
文摘Methane(CH_(4))is a potent greenhouse gas that has a substantial impact on global warming due to its substantial influence on the greenhouse effect.Increasing extreme precipitation events,such as drought,attributable to global warming that caused by greenhouse gases,exert a profound impact on the intricate biological processes associated with CH_(4) uptake.Notably,the timing of extreme drought occurrence emerges as a pivotal factor influencing CH_(4) uptake,even when the degree of drought remains constant.However,it is still unclear how the growing season regulates the response of CH_(4) uptake to extreme drought.In an effort to bridge this knowledge gap,we conducted a field manipulative experiment to evaluate the impact of extreme drought on CH_(4) uptake during early,middle,and late growing stages in a temperate steppe of Inner Mongolia Autonomous Region,China.The result showed that all extreme drought consistently exerted positive effects on CH_(4) uptake regardless of seasonal timing.However,the magnitude of this effect varied depending on the timing of season,as evidenced by a stronger effect in early growing stage than in middle and late growing stages.Besides,the pathways of CH_(4) uptake were different from seasonal timing.Extreme drought affected soil physical-chemical properties and aboveground biomass(AGB),consequently leading to changes in CH_(4) uptake.The structural equation model showed that drought both in the early and middle growing stages enhanced CH_(4) uptake due to reduced soil water content(SWC),leading to a decrease in NO_(3)–-N and an increase in pmoA abundance.However,drought in late growing stage primarily enhanced CH_(4) uptake only by decreasing SWC.Our results suggested that seasonal timing significantly contributed to regulate the impacts of extreme drought pathways and magnitudes on CH_(4) uptake.The findings can provide substantial implications for understanding how extreme droughts affect CH_(4) uptake and improve the prediction of potential ecological consequence under future climate change.
基金supported by the Knowledge InnovationGrant of the Chinese Academy of Sciences (No. kzcx2-yw-418-03)
文摘Zoige wetland is one of the most important methane emission centers in China. The oxidation of methane in the wetland affects global warming, soil ecology and atmospheric chemistry. Despite their global significance, microorganisms that consume methane in Zoige wetland remain poorly characterized. In this study, we investigated methanotrophs diversity in soil samples from both anaerobic site and aerobic site in Zoige wetland using pmoA gene as a molecular marker. The cloning library was constructed according to the pmoA sequences detected. Four clusters of methanotrophs were detected. The phylogenetic tree showed that all four clusters detected were affiliated to type I methanotrophs. Two novel clusters (cluster 1, cluster 2) were found to relate to none of the recognized genera of methanotrophs. These clusters have no cultured representatives and reveal an ecological adaptation of particular uncultured methanotrophs in Zoige wetland. Two clusters were belonging to Methylobacter and Methylococcus separately. Denaturing gradient gel electrophoresis gel bands pattern retrieved from these two samples revealed that the community compositions of anaerobic soil and aerobic soil were different from each other while anaerobic soil showed a higher metanotrophs diversity. Real-time PCR assays of the two samples demonstrated that aerobic soil sample in Zoige wetland was 1.5 times as much copy numbers as anaerobic soil. These data illustrated that methanotrophs are a group of microorganisms influence the methane consumption in Zoige wetland.
基金Supported by the Centro de Investigación y de Estudios Avanzados del IPN,Mexico and the Consejo Nacional de Ciencia y Tecnología,Mexico(Nos.153216,232468 and 245119)
文摘Global warming, as a result of an increase in the mean temperature of the planet, might lead to catastrophic events for humanity. This temperature increase is mainly the result of an increase in the atmospheric greenhouse gases (GHG) concentration. Water vapor, carbon dioxide (CO2), methane (CH4) and nitrous oxide (N20) are the most important GHG, and human activities, such as industry, livestock and agriculture, contribute to the production of these gases. Methane, at an atmospheric concentration of 1.7 gmol tool-1 currently, is responsible for 16% of the global warming due to its relatively high global warming potential. Soils play an important role in the CH4 cycle as methanotrophy (oxidation of CH4) and methanogenesis (production of CH4) take place in them. Understanding methanogenesis and methanotrophy is essential to establish new agriculture techniques and industrial processes that contribute to a better balance of GHG. The current knowledge of methanogenesis and methanotrophy in soils, anaerobic CH4 oxidation and methanotrophy in extreme environments is also discussed.
基金supported by the Knowledge Innovation Program of Chinese Academy of Sciences(No. KSCX2-YW-Z-1020,KZCX2-YW-JC401)the Natural Science Foundation of China(No. 40871129)
文摘Soil samples were collected from three plots under different land utilization patterns including degradation, farming, and restoration. The abundances of methanotrophs were quantified using real-time polymerase chain reaction (PCR) based on the pmoA and 16S rRNA genes, and the community fingerprint was analyzed using denaturing gradient gel electrophoresis (DGGE) aiming at pmoA gene. Significantly lower 16S rRNA and pmoA genes copies were found in the degradation treatment than in farming and restoration. Higher abundances of Type I than those of Type II methanotrophs were detected in all treatments, The treatment of farming was clearly separated from degradation and restoration according to the DGGE profile by cluster analysis. The lowest diversity indices were observed in the F (farming plot), suggesting that the community structure was strongly affected by farming activities. There were significantly positive correlations between the copy numbers of pmoA also Type II-related 16S rRNA genes and soil available K content. Strong negative and positive correlations were found between Type I and soil pH, and available P content, respectively. We concluded that the vegetation cover or not, soil characteristics including pH and nutrients of P and K as a result of anthropogenic disturbance may be key factors affecting methanotrophic communities in upland soil.
基金supported by the Hi-Tech Research and Development Program (863) of China (No. 2007AA06Z349)the Science and Technology Commission of Shanghai Municipality (No. 09DZ2251700)
文摘Aged refuse from waste landfills closed for eight years was examined and found to contain rich methanotrophs capable of biooxidation for methane. Specially, community structure and methane oxidation capability of methanotrophs in the aged refuse were studied. The amount of methanotrophs ranged 61.97×10^3-632.91×10^3 cells/g (in dry basis) in aged refuse from Shanghai Laogang Landfill. Type I and II methanotrophs were found in the aged refuse in the presence of sterilized sewage sludge and only Type I methanotrophs were detected in the presence of nitrate minimal salt medium (NMS). The clone sequences of the pmoA gene obtained from the aged refuse were similar to the pmoA gene of Methylobacter, Methylocaldum, and Methylocystis, and two clones were distinct with known genera of Type I methanotrophs according to phylogenetic analysis. Aged refuse enriched with NMS was used for methane biological oxidation and over 93% conversions were obtained.
基金supported by the National Key Scientific and Technology Project for Water Pollution Treatment of China (No.2012ZX07202006)
文摘Methanotrophic–heterotrophic communities were selectively enriched from sewage sludge to obtain a mixed culture with high levels of poly-β-hydroxybutyrate(PHB)accumulation capacity from methane.Methane was used as the carbon source,N2as sole nitrogen source,and oxygen and Cu content were varied.Copper proved essential for PHB synthesis.All cultures enriched with Cu could accumulate high content of PHB(43.2%–45.9%),while only small amounts of PHB were accumulated by cultures enriched without Cu(11.9%–17.5%).Batch assays revealed that communities grown with Cu and a higher O2content synthesized more PHB,which had a wider optimal CH4:O2range and produced a high PHB content(48.7%)even though in the presence of N2.In all methanotrophic–heterotrophic communities,both methanotrophic and heterotrophic populations showed the ability to accumulate PHB.Although methane was added as the sole carbon source,heterotrophs dominated with abundances between 77.2%and 85.6%.All methanotrophs detected belonged to type II genera,which formed stable communities with heterotrophs of different PHB production capacities.
基金This work is supported by the National Key R&D Program of China(2018YFA0901500)National Natural Science Foundation of China(21878241)and Open Funding Project of the State Key Laboratory of Bioreactor Engineering.
文摘Methanotrophic bacteria are entities with innate biocatalytic potential to biofilter and oxidize methane into simpler compounds concomitantly conserving energy,which can contribute to copious industrial applications.The future and efficacy of such industrial applications relies upon acquiring and/or securing robust methanotrophs with taxonomic and phenotypic diversity.Despite several dramatic advances,isolation of robust methanotrophs is still a long-way challenging task with several lacunae to be filled in sequentially.Methanotrophs with high tolerance to methane can be isolated and cultivated by mimicking natural environs,and adopting strategies like adaptive metabolic evolution.This review summarizes existent and innovative methods for methanotrophic isolation and purification,and their respective applications.A comprehensive description of new insights shedding light upon how to isolate and concomitantly augment robust methanotrophic metabolism in an orchestrated fashion follows.
基金Natural Science and Engineering Research Council of Canada (NSERC)Ontario Center of Excellence (OCE), Canada, Seed Fund
文摘Methanotrophic based process can be the remedy to offset the wastewater treatment facilities increasing energy requirements due to methanotroph's unique ability to integrate methane assimilation with multiple biotechnological applications like biological nitrogen removal and methanol production. Regardless of the methanotrophic process end product, the challenge to maintain stable microbial growth in the methanotrophs cultivation bioreactor at higher cell densities is one of the major obstacles facing the process upscaling. Therefore, a series of consecutive batch tests were performed to attentively investigate the biomass density influence on type I methanotrophs bacterial growth. In addition, food to microorganisms(F/M), carbon to nitrogen(C/N) and nitrogen to microorganisms(N/M) ratio effect on the microbial activity was studied for the first time. It was clarified that the F/M ratio is the most influencing factor on the microbial growth at higher biomass densities rather than the biomass density increase, whereas C/N and N/M ratio change, while using nitrate as the nitrogen source,does not influence methanotrophs microbial growth. These study results would facilitate the scaling up of methanotrophic based biotechnology by identifying that F/M ratio as the key parameter that influences methanotrophs cultivation at high biomass densities.
基金financially supported by the National Natural Science Foundation of China (Nos. 91751204, 41630862, 41701302, 41530857, and 41877062)The first author, Ms. Nasrin Sultana, gratefully acknowledges the Organization for Women in Science for the Developing World (OWSD) Ph.D. Fellowship。
文摘Biological methane oxidation is a crucial process in the global carbon cycle that reduces methane emissions from paddy fields and natural wetlands into the atmosphere.However,soil organic carbon accumulation associated with microbial methane oxidation is poorly understood.Therefore,to investigate methane-derived carbon incorporation into soil organic matter,paddy soils originated from different parent materials(Inceptisol,Entisol,and Alfisol) were collected after rice harvesting from four major rice-producing regions in Bangladesh.Following microcosm incubation with 5%(volume/volume)^(13) CH_(4),soil^(13) C-atom abundances significantly increased from background level of 1.08% to 1.88%–2.78%,leading to a net methane-derived accumulation of soil organic carbon ranging from 120 to 307 mg kg^(-1).Approximately 23.6%–60.0% of the methane consumed was converted to soil organic carbon during microbial methane oxidation.The phylogeny of^(13) C-labeled pmoA(enconding the alpha subunit of the particulate methane monooxygenase) and 16 S rRNA genes further revealed that canonical α(type II) and γ(type I) Proteobacteria were active methane oxidizers.Members within the Methylobacter-and Methylosarcina-affiliated type Ia lineages dominated active methane-oxidizing communities that were responsible for the majority of methane-derived carbon accumulation in all three paddy soils,while Methylocystis-affiliated type IIa lineage was the key contributor in one paddy soil of Inceptisol origin.These results suggest that methanotroph-mediated synthesis of biomass plays an important role in soil organic matter accumulation.This study thus supports the concept that methanotrophs not only consume the greenhouse gas methane but also serve as a key biotic factor in maintaining soil fertility.
基金supported by the National High Technology Research and Development Program (863) of China (No.2007AA06Z350)the State Key Project for National Science and Technology (No. 2006BAC06B05)
文摘A simulated landfill biocover microcosm consisting of a modifying ceramsite material and compost were investigated.Results show that the mixture can improve the material porosity and achieve a stable and highly efficient (100%) methane oxidation over an extended operating period.The diversity of the methanotrophic community in the microcosm was assessed.Type I methanotrophs were enhanced in the microcosm due to the increased air diffusion and distribution,whereas the microbial diversity and population density of type II methanotrophs were not significantly affected.Moreover,the type I methanotrophic community structure significantly varied with the reactor height,whereas that of type II methanotrophic communities did not exhibit a spatial variation.Phylogenetic analysis showed that type I methanotroph-based nested polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) resulted in the detection of eight different populations,most of which are related to Methylobacter sp.,whereas that of type II resulted in the detection of nine different populations,most of which are related to Methylocystaceae.Methanotrophic community analysis also indicated that a number of new methanotrophic genera not closely related to any known methanotrophic populations were present.