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
Background With the global expansion of aquaculture and the increasing demand for fish meal,identifying appropriate and sustainable alternative protein sources for aquafeeds has become essential.Single-cell protein(SC...Background With the global expansion of aquaculture and the increasing demand for fish meal,identifying appropriate and sustainable alternative protein sources for aquafeeds has become essential.Single-cell protein(SCP),derived from methanotrophic bacteria,presents a promising alternative by converting methane into protein,potentially addressing both the need for alternative protein sources and reducing industrial greenhouse gas emissions.This study aimed to evaluate the effects of different levels of SCP inclusion(0%,25%,50%,and 75%fish meal replacement)on the health,gene expression,and gut microbiome of yellowtail kingfish(YTK,Seriola lalandi)following a 35-day growth trial.Results The study found that SCP inclusion at the highest level of fishmeal replacement(75%)induced a mild inflammatory response in the hindgut of the fish.However,micromorphological assessments of the hindgut,serum biochemistry,and gene expression analyses revealed no significant detrimental effects from SCP replacement.Notably,there were indications of improved lipid digestibility with SCP.Furthermore,SCP inclusion significantly enhanced microbial richness and altered the composition of the gut microbiome,introducing beneficial bacterial taxa that may contribute to improved gut health and resilience.Conclusions This study highlights SCP as a viable and sustainable alternative to fish meal in YTK diets.The findings suggest that SCP can be included in YTK diets without adverse health effects at moderate levels and may even offer benefits in terms of lipid digestibility and gut microbiome diversity.These results contribute to the advancement of more sustainable aquaculture practices.展开更多
Anaerobic oxidation of methane(AOM)can contribute to reducing methane emissions in landfills;however,the AOM rates vary depending on the inoculum source.This study addressed the capacity of AOM of a fermentative micro...Anaerobic oxidation of methane(AOM)can contribute to reducing methane emissions in landfills;however,the AOM rates vary depending on the inoculum source.This study addressed the capacity of AOM of a fermentative microbial community derived from a reactor treatingmunicipal solidwastes.First,the inoculum’s autotrophic capacitywas verified using a gasmixture of 75% CO_(2) and 25% H_(2).Results demonstrated that the fermentative microbial community reached amaximum CO_(2) consumption rate of 22.5±1.2 g CO_(2)/(m^(3)·h),obtaining acetate as the main product.Then,the inoculum was grown on a gas mixture of 50%CH_(4),35%CO_(2),and 15%N_(2),using iron(Fe^(3+))as the electron acceptor.The AOM rates increased over time and peaked at 3.1±0.9 g CH_(4)/(m^(3)·h)by 456 h with the simultaneous consumption of CO_(2).Acetate was the main product,with amaximum concentration of 180±9mg/L.By 408 h,a bacterial cluster of indicator species correlated with the AOM rates,including to Rhodobactereceae(r=0.80),Oceanicola(r=0.80),Propionicicella(r=0.77),Christensenellaceae(r=0.58),Oscillospiraceae(r=0.53),Mobilitalea(r=0.66),Hungateiclostridiaceae(r=0.46),and Izemoplasmatales(r=0.77).Methanosarcina,Methanobacterium,and Methanoculleus correlated with the AOM and CO_(2) consumption rates.A co-occurrence network analysis showed that Methanosarcina positively interacted with syntrophic bacteria like Christensenellaceae and Acinetobacter and diverse heterotrophic bacteria.This study demonstrated the feasibility of obtaining a CH_(4)-oxidizing microbial community in 16 days,exhibiting AOM rates higher than those reported for soils.展开更多
Global efforts to avert climate change cannot succeed without tackling the emission of methane from soil and other ecosystems.Methane is a greenhouse gas that retains heat in the atmosphere and causes global warming.I...Global efforts to avert climate change cannot succeed without tackling the emission of methane from soil and other ecosystems.Methane is a greenhouse gas that retains heat in the atmosphere and causes global warming.Its production is the last step of organic matter decomposition,and it is produced by methanogenic archaea bearing the functional gene mcrA(encoding methyl-coenzyme M reductase).Methane production involves the reduction of acetate or carbon dioxide in a microaerophilic or anaerobic environment under the catalytic actions of methyl-coenzyme M to generate methane.On the other hand,methane-oxidizing bacteria,also known as methanotrophs,through the catalytic action of particulate methane monooxygenase(pMMO),oxidize methane and reduce its emission to the atmosphere.In essence,both production and consumption of methane happen within the soil.Methanotrophs and methanogens inhabit the same soil environment.In fact,a shift in the balance between methanogen and methanotroph activities and abundances could influence soil methane emission and global warming.In this review,we highlight recent advances in drivers of methane flux,pmoA(encoding pMMO)and mcrA gene abundances,methane emission and control,relationships between microbial functional gene abundances and soil functions,and methods for studying the pmoA and mcrA gene abundances in soil.We also highlight gaps that need to be filled and the impact of the mcrA/pmoA gene abundance ratio in driving the methane emission rate in soil.We also discuss the various abiotic factors that control pmoA and mcrA gene abundances.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
Wetlands are one of the important natural sources of atmospheric methane (CH_4),as an important part of wetlands,floating plants can be expected to affect methane release.However,the effects of floating plants on meth...Wetlands are one of the important natural sources of atmospheric methane (CH_4),as an important part of wetlands,floating plants can be expected to affect methane release.However,the effects of floating plants on methane release are limited.In this study,methane fluxes,physiochemical properties of the overlying water,methane oxidation potential and rhizospheric bacterial community were investigated in simulated wetlands with floating plants Eichhornia crassipes,Hydrocharis dubia,and Trapa natans.We found that E.crassipes,H.dubia,and T.natans plants could inhibit 84.31%-97.31%,4.98%-88.91%and 43.62%-92.51%of methane fluxes at interface of water-atmosphere compared to Control,respectively.Methane fluxes were negatively related to nutrients concentration in water column but positively related to the aerenchyma proportions of roots,stems,and leaves.At the same biomass,root of E.crassipes (36.44%) had the highest methane oxidation potential,followed by H.dubia (12.99%) and T.natans (11.23%).Forty-five bacterial phyla in total were identified on roots of three plants and 7 bacterial genera (2.10%-3.33%) were known methanotrophs.Type I methanotrophs accounted for 95.07%of total methanotrophs.The pmoA gene abundances ranged from 1.90×10^(16)to 2.30×10^(18)copies/g fresh weight of root biofilms.Abundances of pmoA gene was significantly positively correlated with environmental parameters.Methylotrophy (5.40%) and methanotrophy (3.75%) function were closely related to methane oxidation.This study highlights that floating plant restoration can purify water and promote carbon neutrality partially by reducing methane fluxes through methane oxidation in wetlands.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
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
A new landfill cover system,biochar-methanotrophs-clay(BMC)cover is recommended for reducing methane emissions at landfills.It also contributes to decreasing soil permeability and improving soil water retention in a l...A new landfill cover system,biochar-methanotrophs-clay(BMC)cover is recommended for reducing methane emissions at landfills.It also contributes to decreasing soil permeability and improving soil water retention in a long time,due to highly porous structure of biochar and the growth metabolism of methanotrophs.To determine the effects of biochar content,oxidation aging times and methane-filled days on hydraulic properties,a total of 60 groups of experiments were conducted.The saturated hydraulic conductivity(k_(sat))was obtained by flexible wall permeameter with controllable hydraulic head pressure.The results showed that the k_(sat)of BMC increased with increasing biochar content and oxidation aging times,while decreased with adding methane-filled days.The soil-water characteristic curves(SWCCs)were obtained with soil suction measured by the filter paper method.The results indicated the water retention capacity of MBC reduced with increasing oxidation aging times but increased with adding methane-filled days.Detected by mercury intrusion porosimetry(MIP),fourier transform infrared spectroscopy(FTIR)and scanning electron microscope(SEM),the differences displayed the changes of pore structures and extracellular polymeric substances(EPS).The oxidation aging of biochar increased the volume of pores,resulting in the increased k_(sat)and the decreased water retention capacity.However,the growing of methanotrophs decreased the volume of pores,resulting in the k_(sat)decreased and the water retention capacity increased due to EPS.No matter how many times the oxidation aging process was experienced,the BMC with longer methane-filled days exhibited relatively lower k_(sat)and better water retention capacity.This implied a more stable barrier capacity to reduce water infiltration in the long term.By combing a series of macro and micro experiments,this paper provides theoretical guidance for the application of biochar-methanotroph-clay mixture to landfill covers.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
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
In traditional hydrometeorology and ice physics, when analyzing the development of ice, only degree-days of frost are taken into account. Despite the presence of pores in the ice formed during inhomogeneous crystalliz...In traditional hydrometeorology and ice physics, when analyzing the development of ice, only degree-days of frost are taken into account. Despite the presence of pores in the ice formed during inhomogeneous crystallization and dynamic loads, ice is universally considered monolithic. Situations where ice cannot withstand design loads in frosty conditions are academically inexplicable, although it is common knowledge, for example, porous ice in reeds. Proof of methane accumulations under the ice<span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">-</span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">fountains of fire over holes in sw</span><span style="font-family:Verdana;">ampy waters. “Culprit” is methane. Methanotrophic microorganis</span><span style="font-family:Verdana;">ms, structurally and functionally specialize in using it as a source of carbon and energy, and turn monolithic ice into porous, practically without changing its thi</span><span style="font-family:Verdana;">ckness. When rushing, dark products of methanotrophy and detrit</span><span style="font-family:Verdana;">us raised by gas bubbles appear on the surface of the ice. The albedo of the ice surface decreases, contributing to its melting and the formation of dilutions. The early melting of ice in dilutions and their transformation into vast wormwood is due to methanotrophy products and seismogenic small detritus, which gravitationally slowly settling out of the cold photic layer, thinning it, contributes to an additional insolation warm-up of 2<span style="white-space:nowrap;">°</span>C ÷ 3<span style="white-space:nowrap;">°</span>C, freeing the waters from ice months earlier than normal. <p> <br /> </p> </span></span></span></span>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
To accelerate the efficiency of methane biodegradation in landfills, a Gram-negative, rod-shaped, non-motile, non-spore-forming bacterium, JTA1, which can utilize methane as well as acetate, was isolated from the Laog...To accelerate the efficiency of methane biodegradation in landfills, a Gram-negative, rod-shaped, non-motile, non-spore-forming bacterium, JTA1, which can utilize methane as well as acetate, was isolated from the Laogang MSW landfills, Shanghai, China. Strain JTA1 was a member of genus Methylocystis on the basis of 16S rRNA and pmoA gene sequence similarity. The maximum specific cell growth rates （μmax= 0.042 hr-1, R2= 0.995） was derived through Boltzmann simulation, and the apparent half-saturation constants （Km（app） = 7.08 mmol/L, R2 = 0.982） was calculated according to Michaelis-Menton hyperbolic model, indicating that Methylocystis strain JTA1 had higher-affinity potential for methane oxidation than other reported methanotrophs. By way of adding the strain JTA1 culture, the methane consumption of aged refuse reached 115 mL, almost two times of control experiment. In addition, high tolerance of Methylocystis strain JTA1 to chloroform could facilitate the methane oxidation of aged refuse bio-covers. At the chloroform concentration of 50 mg/L, the methane-oxidation rate of bio-cover reached 0.114 mL/（day.g）, much higher than the highest rate, 0.0135 mL/（day.g）, of reported bio-covers. In conclusion, strain JTA1 opens up a new possibility for environmental biotechnology, such as soil or landfills bioremediation and wastewater decontamination.展开更多
The coupled effects of nitrogen source and methane monooxygenase（MMO） on the growth and poly-β-hydroxybutyrate（PHB） accumulation capacity of methanotrophs were explored.The ammonia-supplied methanotrophs expressi...The coupled effects of nitrogen source and methane monooxygenase（MMO） on the growth and poly-β-hydroxybutyrate（PHB） accumulation capacity of methanotrophs were explored.The ammonia-supplied methanotrophs expressing soluble MMO（s MMO） grew at the highest rate, while N2-fixing bacteria expressing particulate MMO（p MMO） grew at the lowest rate. Further study showed that more hydroxylamine and nitrite was formed by ammonia-supplied bacteria containing p MMO, which might cause their slightly lower growth rate. The highest PHB content（51.0%） was obtained under nitrogen-limiting conditions with the inoculation of nitrate-supplied bacteria containing p MMO. Ammoniasupplied bacteria also accumulated a higher content of PHB（45.2%） with the expression of p MMO, while N2-fixing bacteria containing p MMO only showed low PHB production capacity（32.1%）. The maximal PHB contents of bacteria expressing s MMO were low, with no significant change under different nitrogen source conditions. The low MMO activity,low cell growth rate and low PHB production capacity of methanotrophs continuously cultivated with N2 with the expression of p MMO were greatly improved in the cyclic NO3-N2 cultivation regime, indicating that long-term deficiency of nitrogen sources was detrimental to the activity of methanotrophs expressing pMMO.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
Elevated levels of atmospheric CO_(2)(eCO_(2))promote rice growth and increase methane(CH_(4))emissions from rice paddies,because increased input of plant photosynthate to soil stimulates methanogenic archae.However,t...Elevated levels of atmospheric CO_(2)(eCO_(2))promote rice growth and increase methane(CH_(4))emissions from rice paddies,because increased input of plant photosynthate to soil stimulates methanogenic archae.However,temporal trends in the effects of eCO_(2)on rice growth and CH_(4)emissions are still unclear.To investigate changes in the effects of eCO_(2)over time,we conducted a two-season pot experiment in a walk-in growth chamber.Positive effects of eCO_(2)on rice leaf photosynthetic rate,biomass,and grain yield were similar between growing seasons.However,the effects of eCO_(2)on CH_(4) emissions decreased over time.Elevated CO_(2)increased CH_(4)emissions by 48%-101%in the first growing season,but only by 28%-30%in the second growing season.We also identified the microbial process underlying the acclimation of CH4 emissions to atmospheric CO_(2)enrichment:eCO_(2)stimulated the abundance of methanotrophs more strongly in soils that had been previously exposed to eCO_(2)than in soils that had not been.These results emphasize the need for long-term eCO_(2)experiments for accurate predictions of terrestrial feedbacks.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
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Fisheries Research Development Corporation,Grant/Award Number:2017-030.
文摘Background With the global expansion of aquaculture and the increasing demand for fish meal,identifying appropriate and sustainable alternative protein sources for aquafeeds has become essential.Single-cell protein(SCP),derived from methanotrophic bacteria,presents a promising alternative by converting methane into protein,potentially addressing both the need for alternative protein sources and reducing industrial greenhouse gas emissions.This study aimed to evaluate the effects of different levels of SCP inclusion(0%,25%,50%,and 75%fish meal replacement)on the health,gene expression,and gut microbiome of yellowtail kingfish(YTK,Seriola lalandi)following a 35-day growth trial.Results The study found that SCP inclusion at the highest level of fishmeal replacement(75%)induced a mild inflammatory response in the hindgut of the fish.However,micromorphological assessments of the hindgut,serum biochemistry,and gene expression analyses revealed no significant detrimental effects from SCP replacement.Notably,there were indications of improved lipid digestibility with SCP.Furthermore,SCP inclusion significantly enhanced microbial richness and altered the composition of the gut microbiome,introducing beneficial bacterial taxa that may contribute to improved gut health and resilience.Conclusions This study highlights SCP as a viable and sustainable alternative to fish meal in YTK diets.The findings suggest that SCP can be included in YTK diets without adverse health effects at moderate levels and may even offer benefits in terms of lipid digestibility and gut microbiome diversity.These results contribute to the advancement of more sustainable aquaculture practices.
基金This work was supported by the DGAPA-UNAM(PAPIIT project,No.IN102721)the support from CONAHCYT through the Investigadoras e Investigadores por Mexico program(Researcher ID 6407,Project 265).
文摘Anaerobic oxidation of methane(AOM)can contribute to reducing methane emissions in landfills;however,the AOM rates vary depending on the inoculum source.This study addressed the capacity of AOM of a fermentative microbial community derived from a reactor treatingmunicipal solidwastes.First,the inoculum’s autotrophic capacitywas verified using a gasmixture of 75% CO_(2) and 25% H_(2).Results demonstrated that the fermentative microbial community reached amaximum CO_(2) consumption rate of 22.5±1.2 g CO_(2)/(m^(3)·h),obtaining acetate as the main product.Then,the inoculum was grown on a gas mixture of 50%CH_(4),35%CO_(2),and 15%N_(2),using iron(Fe^(3+))as the electron acceptor.The AOM rates increased over time and peaked at 3.1±0.9 g CH_(4)/(m^(3)·h)by 456 h with the simultaneous consumption of CO_(2).Acetate was the main product,with amaximum concentration of 180±9mg/L.By 408 h,a bacterial cluster of indicator species correlated with the AOM rates,including to Rhodobactereceae(r=0.80),Oceanicola(r=0.80),Propionicicella(r=0.77),Christensenellaceae(r=0.58),Oscillospiraceae(r=0.53),Mobilitalea(r=0.66),Hungateiclostridiaceae(r=0.46),and Izemoplasmatales(r=0.77).Methanosarcina,Methanobacterium,and Methanoculleus correlated with the AOM and CO_(2) consumption rates.A co-occurrence network analysis showed that Methanosarcina positively interacted with syntrophic bacteria like Christensenellaceae and Acinetobacter and diverse heterotrophic bacteria.This study demonstrated the feasibility of obtaining a CH_(4)-oxidizing microbial community in 16 days,exhibiting AOM rates higher than those reported for soils.
文摘Global efforts to avert climate change cannot succeed without tackling the emission of methane from soil and other ecosystems.Methane is a greenhouse gas that retains heat in the atmosphere and causes global warming.Its production is the last step of organic matter decomposition,and it is produced by methanogenic archaea bearing the functional gene mcrA(encoding methyl-coenzyme M reductase).Methane production involves the reduction of acetate or carbon dioxide in a microaerophilic or anaerobic environment under the catalytic actions of methyl-coenzyme M to generate methane.On the other hand,methane-oxidizing bacteria,also known as methanotrophs,through the catalytic action of particulate methane monooxygenase(pMMO),oxidize methane and reduce its emission to the atmosphere.In essence,both production and consumption of methane happen within the soil.Methanotrophs and methanogens inhabit the same soil environment.In fact,a shift in the balance between methanogen and methanotroph activities and abundances could influence soil methane emission and global warming.In this review,we highlight recent advances in drivers of methane flux,pmoA(encoding pMMO)and mcrA gene abundances,methane emission and control,relationships between microbial functional gene abundances and soil functions,and methods for studying the pmoA and mcrA gene abundances in soil.We also highlight gaps that need to be filled and the impact of the mcrA/pmoA gene abundance ratio in driving the methane emission rate in soil.We also discuss the various abiotic factors that control pmoA and mcrA gene abundances.
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supported by the National Natural Science Foundation of China (No. 51879084)the Technology Program for Water Pollution Control and Treatment of China (No. 2018ZX07208-004)。
文摘Wetlands are one of the important natural sources of atmospheric methane (CH_4),as an important part of wetlands,floating plants can be expected to affect methane release.However,the effects of floating plants on methane release are limited.In this study,methane fluxes,physiochemical properties of the overlying water,methane oxidation potential and rhizospheric bacterial community were investigated in simulated wetlands with floating plants Eichhornia crassipes,Hydrocharis dubia,and Trapa natans.We found that E.crassipes,H.dubia,and T.natans plants could inhibit 84.31%-97.31%,4.98%-88.91%and 43.62%-92.51%of methane fluxes at interface of water-atmosphere compared to Control,respectively.Methane fluxes were negatively related to nutrients concentration in water column but positively related to the aerenchyma proportions of roots,stems,and leaves.At the same biomass,root of E.crassipes (36.44%) had the highest methane oxidation potential,followed by H.dubia (12.99%) and T.natans (11.23%).Forty-five bacterial phyla in total were identified on roots of three plants and 7 bacterial genera (2.10%-3.33%) were known methanotrophs.Type I methanotrophs accounted for 95.07%of total methanotrophs.The pmoA gene abundances ranged from 1.90×10^(16)to 2.30×10^(18)copies/g fresh weight of root biofilms.Abundances of pmoA gene was significantly positively correlated with environmental parameters.Methylotrophy (5.40%) and methanotrophy (3.75%) function were closely related to methane oxidation.This study highlights that floating plant restoration can purify water and promote carbon neutrality partially by reducing methane fluxes through methane oxidation in wetlands.
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National Natural Science Foundation of China(Grant No.41977214)the Research Platform Open Fund Project of Zhejiang Industry and Trade Vocation College(Kf202202).
文摘A new landfill cover system,biochar-methanotrophs-clay(BMC)cover is recommended for reducing methane emissions at landfills.It also contributes to decreasing soil permeability and improving soil water retention in a long time,due to highly porous structure of biochar and the growth metabolism of methanotrophs.To determine the effects of biochar content,oxidation aging times and methane-filled days on hydraulic properties,a total of 60 groups of experiments were conducted.The saturated hydraulic conductivity(k_(sat))was obtained by flexible wall permeameter with controllable hydraulic head pressure.The results showed that the k_(sat)of BMC increased with increasing biochar content and oxidation aging times,while decreased with adding methane-filled days.The soil-water characteristic curves(SWCCs)were obtained with soil suction measured by the filter paper method.The results indicated the water retention capacity of MBC reduced with increasing oxidation aging times but increased with adding methane-filled days.Detected by mercury intrusion porosimetry(MIP),fourier transform infrared spectroscopy(FTIR)and scanning electron microscope(SEM),the differences displayed the changes of pore structures and extracellular polymeric substances(EPS).The oxidation aging of biochar increased the volume of pores,resulting in the increased k_(sat)and the decreased water retention capacity.However,the growing of methanotrophs decreased the volume of pores,resulting in the k_(sat)decreased and the water retention capacity increased due to EPS.No matter how many times the oxidation aging process was experienced,the BMC with longer methane-filled days exhibited relatively lower k_(sat)and better water retention capacity.This implied a more stable barrier capacity to reduce water infiltration in the long term.By combing a series of macro and micro experiments,this paper provides theoretical guidance for the application of biochar-methanotroph-clay mixture to landfill covers.
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 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.
文摘In traditional hydrometeorology and ice physics, when analyzing the development of ice, only degree-days of frost are taken into account. Despite the presence of pores in the ice formed during inhomogeneous crystallization and dynamic loads, ice is universally considered monolithic. Situations where ice cannot withstand design loads in frosty conditions are academically inexplicable, although it is common knowledge, for example, porous ice in reeds. Proof of methane accumulations under the ice<span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">-</span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">fountains of fire over holes in sw</span><span style="font-family:Verdana;">ampy waters. “Culprit” is methane. Methanotrophic microorganis</span><span style="font-family:Verdana;">ms, structurally and functionally specialize in using it as a source of carbon and energy, and turn monolithic ice into porous, practically without changing its thi</span><span style="font-family:Verdana;">ckness. When rushing, dark products of methanotrophy and detrit</span><span style="font-family:Verdana;">us raised by gas bubbles appear on the surface of the ice. The albedo of the ice surface decreases, contributing to its melting and the formation of dilutions. The early melting of ice in dilutions and their transformation into vast wormwood is due to methanotrophy products and seismogenic small detritus, which gravitationally slowly settling out of the cold photic layer, thinning it, contributes to an additional insolation warm-up of 2<span style="white-space:nowrap;">°</span>C ÷ 3<span style="white-space:nowrap;">°</span>C, freeing the waters from ice months earlier than normal. <p> <br /> </p> </span></span></span></span>
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 National Natural Science Foundation of China (No. 51008322)the Natural Science Foundation Project of Chongqing (No. CSTC 2010BB7300,5309)the State Key Laboratory of Pollution Control and Resource Reuse (No. PCRRF10001)
文摘To accelerate the efficiency of methane biodegradation in landfills, a Gram-negative, rod-shaped, non-motile, non-spore-forming bacterium, JTA1, which can utilize methane as well as acetate, was isolated from the Laogang MSW landfills, Shanghai, China. Strain JTA1 was a member of genus Methylocystis on the basis of 16S rRNA and pmoA gene sequence similarity. The maximum specific cell growth rates （μmax= 0.042 hr-1, R2= 0.995） was derived through Boltzmann simulation, and the apparent half-saturation constants （Km（app） = 7.08 mmol/L, R2 = 0.982） was calculated according to Michaelis-Menton hyperbolic model, indicating that Methylocystis strain JTA1 had higher-affinity potential for methane oxidation than other reported methanotrophs. By way of adding the strain JTA1 culture, the methane consumption of aged refuse reached 115 mL, almost two times of control experiment. In addition, high tolerance of Methylocystis strain JTA1 to chloroform could facilitate the methane oxidation of aged refuse bio-covers. At the chloroform concentration of 50 mg/L, the methane-oxidation rate of bio-cover reached 0.114 mL/（day.g）, much higher than the highest rate, 0.0135 mL/（day.g）, of reported bio-covers. In conclusion, strain JTA1 opens up a new possibility for environmental biotechnology, such as soil or landfills bioremediation and wastewater decontamination.
基金supported by the National Key Scientific and Technology Project for Water Pollution Treatment of China(No.2012ZX07202006)the National Natural Science Foundation of China(No.21477014)
文摘The coupled effects of nitrogen source and methane monooxygenase（MMO） on the growth and poly-β-hydroxybutyrate（PHB） accumulation capacity of methanotrophs were explored.The ammonia-supplied methanotrophs expressing soluble MMO（s MMO） grew at the highest rate, while N2-fixing bacteria expressing particulate MMO（p MMO） grew at the lowest rate. Further study showed that more hydroxylamine and nitrite was formed by ammonia-supplied bacteria containing p MMO, which might cause their slightly lower growth rate. The highest PHB content（51.0%） was obtained under nitrogen-limiting conditions with the inoculation of nitrate-supplied bacteria containing p MMO. Ammoniasupplied bacteria also accumulated a higher content of PHB（45.2%） with the expression of p MMO, while N2-fixing bacteria containing p MMO only showed low PHB production capacity（32.1%）. The maximal PHB contents of bacteria expressing s MMO were low, with no significant change under different nitrogen source conditions. The low MMO activity,low cell growth rate and low PHB production capacity of methanotrophs continuously cultivated with N2 with the expression of p MMO were greatly improved in the cyclic NO3-N2 cultivation regime, indicating that long-term deficiency of nitrogen sources was detrimental to the activity of methanotrophs expressing pMMO.
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supported by the National Key Research and Development Program of China(2017YFD0300104,2016YFD0300903,2015BAC02B02)the National Natural Science Foundation of China(32022061)+3 种基金the Special Fund for Agroscientific Research in the Public Interest(201503118,201503122)the Agricultural Science and Technology Innovation Program of CAAS(Y2016PT12,Y2016XT01)the Modern Agricultural Development of Jiangsu Province(2019-SJ-039-07)the GEF Project of Climate Smart Staple Crop Production in China(P144531)。
文摘Elevated levels of atmospheric CO_(2)(eCO_(2))promote rice growth and increase methane(CH_(4))emissions from rice paddies,because increased input of plant photosynthate to soil stimulates methanogenic archae.However,temporal trends in the effects of eCO_(2)on rice growth and CH_(4)emissions are still unclear.To investigate changes in the effects of eCO_(2)over time,we conducted a two-season pot experiment in a walk-in growth chamber.Positive effects of eCO_(2)on rice leaf photosynthetic rate,biomass,and grain yield were similar between growing seasons.However,the effects of eCO_(2)on CH_(4) emissions decreased over time.Elevated CO_(2)increased CH_(4)emissions by 48%-101%in the first growing season,but only by 28%-30%in the second growing season.We also identified the microbial process underlying the acclimation of CH4 emissions to atmospheric CO_(2)enrichment:eCO_(2)stimulated the abundance of methanotrophs more strongly in soils that had been previously exposed to eCO_(2)than in soils that had not been.These results emphasize the need for long-term eCO_(2)experiments for accurate predictions of terrestrial feedbacks.
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.
