A two-year field experiment conducted under dryland conditions in semi-humid and drought-prone regions of China aimed to assess the effect of ammonia-oxidizing bacterial on maize water use efficiency and yield.A heter...A two-year field experiment conducted under dryland conditions in semi-humid and drought-prone regions of China aimed to assess the effect of ammonia-oxidizing bacterial on maize water use efficiency and yield.A heterotrophic ammonia-oxidizing bacteria(HAOB)strain S2_8_1 was used.Six treatments were applied:(1)no irrigation+HAOB strain(DI),(2)no irrigation+blank culture medium(DM),(3)no irrigation control(DCK),(4)irrigation+HAOB(WI),(5)irrigation+blank culture medium(WM),and(6)irrigation control(WCK).Results revealed that HAOB treatment increased maize growth,yield,and water use efficiency over controls,regardless of whether the year was wet or dry.This improvement was attributed to the accelerated nitrification in the rhizosphere soil due to HAOB inoculation,which subsequently led to increased levels of leaf cytokinins.Overall,these findings suggest that HAOB inoculation holds promise as a strategy to boost water use efficiency and maize productivity in dryland agriculture.展开更多
Acetochlor is an increasingly used herbicide on corn in North China. Currently, the effect of acetochlor on soil ammonia-oxidizing bacteria (AOB) communities is not well documented. Here, we studied the diversity and ...Acetochlor is an increasingly used herbicide on corn in North China. Currently, the effect of acetochlor on soil ammonia-oxidizing bacteria (AOB) communities is not well documented. Here, we studied the diversity and community composition of AOB in soil amended with three concentrations of acetochlor (50, 150, 250 mg/kg) and the control (0 mg acetochlor/kg soil) in a microcosm experiment by PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and the phylogenetic analysis of excised ...展开更多
A molecular biology method, fluorescent in situ hybridization(FISH), in which the pre-treatment was improved in allusion to the media of the constructed wetlands(CW), e.g. the soil and the grit, was used to invest...A molecular biology method, fluorescent in situ hybridization(FISH), in which the pre-treatment was improved in allusion to the media of the constructed wetlands(CW), e.g. the soil and the grit, was used to investigate the vertical distribution characteristics of ammonia-oxidizing bacteria(AOB) quantity and the relation with oxidation-reduction potential(ORP) in the Typha latifolia constructed wetlands under three different Ioadings in summer from May to September. Results showed that the quantity of the AOB decreased in the Typha latifolia CW with the increase of vertical depth. However, the AOB quantity was 2-4 times the quantity of the control in the root area. Additionally, ORP in the rhizosphere was found to be higher than other areas, which showed that Typha latifolia CW was in an aerobic state in summer when using simulated non-point sewage at the rural area of Taihu Lake in China and small town combined sewage.展开更多
ObjectiveThe aim was to understand the effects of transgenic DREB soybean on the ammonia-oxidizing bacteria. MethodThe diversity of the cto gene in pot-planted transgenic soybean and near-isogenic non-transgenic soybe...ObjectiveThe aim was to understand the effects of transgenic DREB soybean on the ammonia-oxidizing bacteria. MethodThe diversity of the cto gene in pot-planted transgenic soybean and near-isogenic non-transgenic soybean under normal water condition and drought stress was analyzed by PCR-DGGE and sequence analysis. ResultRhizosphere community diversity of ammonia-oxidizing bacteria showed no difference between the treatments of transgenic soybean and its non-transgenic isolines, moreover transgenic soybean under normal water condition and drought stress improved the diversity of the ammonia-oxidizing bacteria in the harvest time. The phylogenetic analysis revealed that all the sequences of excised DGGE bands were closely related to members of the genus Nitrosovibrio and Nitrosospira of the β-subclass Proteobacteria. ConclusionTransgenic DREB soybean has no adverse impact on soil ammonia-oxidizing bacteria.展开更多
We investigated the communities of ammonia-oxidizing bacteria (AOB) in activated sludge collected from eight wastewater treatment systems using polymerase chain reaction (PCR) followed by terminal restriction frag...We investigated the communities of ammonia-oxidizing bacteria (AOB) in activated sludge collected from eight wastewater treatment systems using polymerase chain reaction (PCR) followed by terminal restriction fragment length polymorphism (T-RFLP), cloning, and sequencing of the α-subunit of the ammonia monooxygenase gene (amoA). The T-RFLP fingerprint analyses showed that different wastewater treatment systems harbored distinct AOB communities. However, there was no remarkable difference among the AOB T- RFLP profiles from different parts of the same system. The T-RFLP fingerprints showed that a full-scale wastewater treatment plant (WWTP) contained a larger number of dominant AOB species than a pilot-scale reactor. The source of influent affected the AOB community, and the WWTPs treating domestic wastewater contained a higher AOB diversity than those receiving mixed domestic and industrial wastewater. However, the AOB community structure was little affected by the treatment process in this study. Phylogenetic analysis of the cloned amoA genes clearly indicated that all the dominant AOB in the systems was closely related to Nitrosomonas spp. not to Nitrosospira spp. Members of the Nitrosomonas oligotropha and Nitrosomonas communis clusters were found in all samples, while members of Nitrosomonas europaea cluster occurred in some systems.展开更多
The spatial distribution of ammonia-oxidizing Betaproteobacteria (βAOB) was investigated by FISH (fluorescence in situ hybridization) and DGGE (denaturing gradient get electrophoresis) techniques in the sedimen...The spatial distribution of ammonia-oxidizing Betaproteobacteria (βAOB) was investigated by FISH (fluorescence in situ hybridization) and DGGE (denaturing gradient get electrophoresis) techniques in the sediment off the Changjiang River Estuary. Sediment samples were collected from eight stations in June before the formation of hypoxia zone in 2006. The abundance of βAOB ranged from 1.87× 10^5 to 3.53×10^5 cells/g of sediment. βAOB abundance did not present a negative correlation with salinity, whereas salinity was implicated as the primary factor affecting nitrification rates. The DGGE profiles of PCR-amplified amoA gene fragments revealed that the βAOB community structure of sample S2 separated from other samples at the level of 40% similarity. The variations in composition ofβAOB were significantly correlated with the salinity, temperature, absorption ability of sediments and TOC. The statistical analysis indicates that theβAOB abundance was a main factor to influence nitrification rates with an influence ratio of 87.7% at the level of 40% biodiversity similarity. Considering the good correlation between βAOB abundance and nitrification estimates, the abundance and diversity of βAOB community could be expected as an indirect index of nitrification activity at the study sea area in summer.展开更多
We investigated the changes in communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira during the operation of a pufferfish Takifugu rubripes recirculating aquaculture system by using high-throughput DNA seq...We investigated the changes in communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira during the operation of a pufferfish Takifugu rubripes recirculating aquaculture system by using high-throughput DNA sequencing.Differences in bacterial communities were observed at days 1-32,47-62 and 78-93 of biofilm development by using 16S rRNA gene pyrosequencing.The relative abundance of Proteobacteria(Gammaproteobacteria)increased,while that of Bacteroidetes(Flavobacteria)decreased.The proportions of Nitrosomonas and Nitrospina ranged from 0.02%to 0.30%and from 0.02%to 0.83%,respectively.Ammonia monooxygenase gene pyrosequencing revealed that the top three operational taxonomic units were related to Nitrosomonas aestuarii(17.5%-61.1%),uncultured beta proteobacterium clone B67S-54(1.9%-45.2%),and uncultured bacterium clone AZPa8(3.6%-24.7%).Nitrite oxidoreductase gene pyrosequencing revealed that the relative abundance of the dominant strain Nitrospira sp.Ecomares 2.1 increased,but that of the abundant species Nitrospira marina decreased.Our results demonstrated that the communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira were changing during the operation of the pufferfish recirculating aquaculture system.展开更多
In order to explore the nitrogen removal process in constructed wetlands(CW s),the moisture,ammonia nitrogen(NH4+-N),nitrate nitrogen(NO3"-N)and nitrification intensity in three wetland plant rhizosphere soils(Ac...In order to explore the nitrogen removal process in constructed wetlands(CW s),the moisture,ammonia nitrogen(NH4+-N),nitrate nitrogen(NO3"-N)and nitrification intensity in three wetland plant rhizosphere soils(Acorns calamus,Typha orientalis,Iris pseudacorus)were investigated at a relatively normal temperature range of15to25The relative abundance of ammonia-oxidizing bacteria(AOB)and ammonia-oxidizing archaea(AOA)were also achieved using fluorescence in situ hybridization(FISH).It is found that T.orientalis achieves the highest nitrification intensity of2.03m g(h?kg)while the second is I.pseudacorrs(1.74m g/(h?kg)),and followed by A.calamus(1.65m g/(h?kg))throughout the experiment.FISH reveals that the abundance of bacteria(1010g_1wet soil)is higher than that of archaea(109g_1wet soil),and AOBare the dominant bacteria in the ammonia oxidation process.The abundance of AOB in te rhizosphere soils from high to low T.orientalis(1.88x1010g"1),I pseudacorus(1.23x1010g1),A.calamus(5.07x109g"1)while the abundance of AOA from high to low ae I.pseudacorus(4.00x109g1),A.calamus(3.52x109g"1),T.orientalis(3.48x109g"1).The study provides valuable evidence of plant selection for nitrogen removal in CWs.展开更多
In this study, we investigated the potential nitrification and community structure of soil-based ammonia-oxidizing bacteria (AOB) in apple orchard soil during different growth periods and explored the effects of env...In this study, we investigated the potential nitrification and community structure of soil-based ammonia-oxidizing bacteria (AOB) in apple orchard soil during different growth periods and explored the effects of environmental factors on nitrification activity and AOB community composition in the soil of a Hanfu apple orchard, using a culture-dependent technique and denaturing gradient gel electrophoresis (DGGE). We observed that nitrification activity and AOB abundance were the highest in November, lower in May, and the lowest in July. The results of statistical analysis indicated that total nitrogen (N) content, NH4+-N content, NO3-N content, and pH showed significant correlations with AOB abundance and nitrification activity in soil. The Shannon-Winner diversity, as well as species richness and evenness indices (determined by PCR-DGGE banding patterns) in soil samples were the highest in September, but the lowest in July, when compared to additional sampled dates. The DGGE fingerprints of soil-based 16S rRNA genes in November were apparently distinct from those observed in May, July, and September, possessing the lowest species richness indices and the highest dominance indices among all four growth periods. Fourteen DGGE bands were excised for sequencing. The resulting analysis indicated that all AOB communities belonged to the 13-Proteobacteria phylum, with the dominant AOB showing high similarity to the Nitrosospira genus. Therefore, soil-based environmental factors, such as pH variation and content of NHa+-N and NO3--N, can substantially influence the abundance of AOB communities in soil, and play a critical role in soil-based nitrification kinetics.展开更多
Subterranean estuaries(STEs)are characterized by the mixing of terrestrial fresh groundwater and seawater in coastal aquifers.Although microorganisms are important components of coastal groundwater ecosystems and play...Subterranean estuaries(STEs)are characterized by the mixing of terrestrial fresh groundwater and seawater in coastal aquifers.Although microorganisms are important components of coastal groundwater ecosystems and play critical roles in biogeochemical transformations in STEs,limited information is available about how their community dynamics interact with hydrological,geochemical and environmental characteristics in STEs.Here,we studied bacterial and archaeal diversities and distributions with 16S rRNA-based Illumina MiSeq sequencing technology between surface water and groundwater in a karstic STE.Principal-coordinate analysis found that the bacterial and archaeal communities in the areas where algal blooms occurred were significantly separated from those in other stations without algal bloom occurrence.Canonical correspondence analysis showed that nutrients and salinity can explain the patterns of bacterial and archaeal community dynamics.The results suggest that hydrological,geochemical and environmental characteristics between surface water and groundwater likely control the bacterial and archaeal diversities and distributions in STEs.Furthermore,we found that some key species can utilize terrestrial pollutants such as nitrate and ammonia in STEs,indicating that these species(e.g.,Nitrosopumilus maritimus,Limnohabitans parvus and Simplicispira limi)may be excellent candidates for in situ degradation/remediation of coastal groundwater contaminations concerned with the nitrate and ammonia.Overall,this study reveals the coupling relationship between the microbial communities and hydrochemical environments in STEs,and provides a perspective of in situ degradation/remediation for coastal groundwater quality management.展开更多
Ammonia oxidation, the first and rate-limiting step of nitrification, is carried out by both ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA). However, the relative importance of AOB and AOA to...Ammonia oxidation, the first and rate-limiting step of nitrification, is carried out by both ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA). However, the relative importance of AOB and AOA to nitrification in terrestrial ecosystems is not well understood. The aim of this study was to investigate the effect of the nitrogen input amount on abundance and community composition of AOB and AOA in red paddy soil. Soil samples of 10-20 cm (root layer soil) and 0-5 cm (surface soil) depths were taken from a red paddy. Rice in the paddy was fertilized with different rates of N as urea of N1 (75 kg N ha" yr-1), N2 (150 kg N ha~ yrl), N3 (225 kg N ha1 yrl) and CK (without fertilizers) in 2009, 2010 and 2011. Abundance and community composition of ammonia oxidizers was analyzed by real-time PCR and denaturing gradient gel electrophoresis (DGGE) based on amoA (the unit A of ammonia monooxygenase) gene. Archaeal amoA copies in N3 and N2 were significantly (P〈0.05) higher than those in CK and N1 in root layer soil or in surface soil under tillering and heading stages of rice, while the enhancement in bacterial amoA gene copies with increasing of N fertilizer rates only took on in root layer soil. N availability and soil NO3--N content increased but soil NH4+-N content didn't change with increasing of N fertilizer rates. Otherwise, the copy numbers of archaeal amoA gene were higher (P〈0.05) than those of bacterial amoA gene in root lary soil or in surface soil. Redundancy discriminate analysis based on DGGE bands showed that there were no obvious differs in composition of AOA or AOB communities in the field among different N fertilizer rates. Results of this study suggested that the abundance of ammonia-oxidizers had active response to N fertilizer rates and the response of AOA was more obvious than that of AOB. Similarity in the community composition of AOA or AOB among different N fertilizer rates indicate that the community composition of ammonia-oxidizers was relatively stable in the paddy soil at least in short term for three years.展开更多
Ammonia-oxidizing archaea(AOA),ammonia-oxidizing bacteria(AOB),and anaerobic ammonia-oxidation(anammox)bacteria are very important contributors to nitrogen cycling in natural environments.Functional gene abundances of...Ammonia-oxidizing archaea(AOA),ammonia-oxidizing bacteria(AOB),and anaerobic ammonia-oxidation(anammox)bacteria are very important contributors to nitrogen cycling in natural environments.Functional gene abundances of these microbes were believed to be well relevant to N-cycling in groundwater systems,especially in the Pearl River Delta(PRD)groundwater with unique high intrinsic ammonia concentrations.In this research,20 sediment samples from two in the PRD were collected for porewater chemistry analysis and quantification of N-cycling related genes,including archaeal and bacterial amoA gene and anammox 16S ribosomal Ribonucleic Acid(rRNA)gene.Quantitative Polymerase Chain Reaction(qPCR)results showed that gene abundances of AOA,AOB,and anammox bacteria ranged from 3.13×10^(5)to 3.21×10^(7),1.83×10^(4)to 2.74×10^(6),and 9.27×10^(4)to 8.96×10^(6)copies/g in the sediment of the groundwater system,respectively.Anammox bacteria and AOA dominated in aquitards and aquifers,respectively,meanwhile,the aquitard-aquifer interfaces were demonstrated as ammonium-oxidizing hotspots in the aspect of gene numbers.Gene abundances of nitrifiers were analyzed with geochemistry profiles.Correlations between gene numbers and environmental variables indicated that the gene abundances were impacted by hydrogeological conditions,and microbial-derived ammonium loss was dominated by AOA in the northwest PRD and by anammox bacteria in the southeast PRD.展开更多
The abundance of ammonia-oxidizing bacteria and archaea and their amo A genes from the aerobic activated sludge tanks,recycled sludge and anaerobic digesters of a full-scale wastewater treatment plant(WWTP)was determi...The abundance of ammonia-oxidizing bacteria and archaea and their amo A genes from the aerobic activated sludge tanks,recycled sludge and anaerobic digesters of a full-scale wastewater treatment plant(WWTP)was determined.Polymerase chain reaction and denaturing gradient gel electrophoresis were used to generate diversity profiles,which showed that each population had a consistent profile although the abundance of individual members varied.In the aerobic tanks,the ammonia-oxidizing bacterial(AOB)population was more than 350 times more abundant than the ammonia-oxidizing archaeal(AOA)population,however in the digesters,the AOA population was more than 10 times more abundant.Measuring the activity of the amo A gene expression of the two populations using RT-PCR also showed that the AOA amo A gene was more active in the digesters than in the activated sludge tanks.Using batch reactors and dd PCR,amo A activity could be measured and it was found that when the AOB amo A activity was inhibited in the anoxic reactors,the expression of the AOA amo A gene increased fourfold.This suggests that these two populations may have a cooperative relationship for the oxidation of ammonia.展开更多
Autotrophic ammonia-oxidizing bacteria (AOB) have been widely studied in constructed wetlands systems, while mixotrophic AOB have been less thoroughly examined. Heterotrophic bacteria were isolated from wastewater a...Autotrophic ammonia-oxidizing bacteria (AOB) have been widely studied in constructed wetlands systems, while mixotrophic AOB have been less thoroughly examined. Heterotrophic bacteria were isolated from wastewater and rhizospheres of macrophytes of constructed wetlands, and then cultivated in a mixotrophic medium containing ammonium and acetic acid. A molecular characterization was accomplished using ITS-PCR amplification, and phylogenetic analysis based on 16S rRNA gene Sequences. Results showed the presence of 35 bacteria, among 400 initially heterotrophic isolates, that were able to remove ammonia. These 35 isolates were classified into 10 genetically different groups based on ITS pattern. Then, a collection of 10 isolates were selected because of their relatively high ammonia removal efficiencies (ARE≥ 80%) and their phylogenetic diversity. In conditions of mixotrophy, these strains were shown to be able to grow (increase of optical density OD660 during incubation with assimilation of nitrogen into cellular biomass) and to oxidize ammonia (important ammonia oxidation efficiencies, AOE between 79% and 87%). Among these facultative mixotrophic AOB, four isolates were genetically related to Firmicutes (Bacillus and Exiguobacterium), three isolates were affiliated to Actinobacteria (Arthrobacter) and three other isolates were associated with Proteobacteria (Pseudomonas, Ochrobactrum and Bordetella).展开更多
Ammonia-oxidizing bacteria(AOB)and archaea(AOA)are two microbial groups mediating nitrification,yet little is presently known about their abundances and community structures at the transcriptional level in wastewater ...Ammonia-oxidizing bacteria(AOB)and archaea(AOA)are two microbial groups mediating nitrification,yet little is presently known about their abundances and community structures at the transcriptional level in wastewater treatment systems(WWTSs).This is a significant issue,as the numerical abundance of AOA or AOB at the gene level may not necessarily represent their functional role in ammonia oxidation.Using amo A genes as molecular markers,this study investigated the transcriptional abundance and community structure of active AOA and AOB in 14 WWTSs.Quantitative PCR results indicated that the transcriptional abundances of AOB amo A(averaged:1.6×10^(8)copies g^(-1)dry sludge)were higher than those of AOA(averaged:3.4×10^(7)copies g^(-1)dry sludge)in all WWTSs despite several higher abundances of AOA amo A at the gene level.Moreover,phylogenetic analysis demonstrated that Nitrosomonas europaea and unknown clusters accounted for 37.66%and 49.96%of the total AOB amo A transcripts,respectively,suggesting their dominant role in driving ammonia oxidation.Meanwhile,AOA amo A transcripts were only successfully retrieved from 3 samples,and the Nitrosospaera sister cluster dominated,accounting for 83.46%.Finally,the substrate utilization kinetics of different AOA and AOB species might play a fundamental role in shaping their niche differentiation,community composition,and functional activity.This study provides a basis for evaluating the relative contributions of ammonia-oxidizing microorganisms(AOMs)to nitrogen conversions in WWTSs.展开更多
基金supported by the National Natural Science Foundation of China(U1304326)the Natural Science Foundation of Henan Provincial(242300421242)。
文摘A two-year field experiment conducted under dryland conditions in semi-humid and drought-prone regions of China aimed to assess the effect of ammonia-oxidizing bacterial on maize water use efficiency and yield.A heterotrophic ammonia-oxidizing bacteria(HAOB)strain S2_8_1 was used.Six treatments were applied:(1)no irrigation+HAOB strain(DI),(2)no irrigation+blank culture medium(DM),(3)no irrigation control(DCK),(4)irrigation+HAOB(WI),(5)irrigation+blank culture medium(WM),and(6)irrigation control(WCK).Results revealed that HAOB treatment increased maize growth,yield,and water use efficiency over controls,regardless of whether the year was wet or dry.This improvement was attributed to the accelerated nitrification in the rhizosphere soil due to HAOB inoculation,which subsequently led to increased levels of leaf cytokinins.Overall,these findings suggest that HAOB inoculation holds promise as a strategy to boost water use efficiency and maize productivity in dryland agriculture.
基金the Natural Science Foun-dation for Young Scientists of China (No. 40701088)
文摘Acetochlor is an increasingly used herbicide on corn in North China. Currently, the effect of acetochlor on soil ammonia-oxidizing bacteria (AOB) communities is not well documented. Here, we studied the diversity and community composition of AOB in soil amended with three concentrations of acetochlor (50, 150, 250 mg/kg) and the control (0 mg acetochlor/kg soil) in a microcosm experiment by PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and the phylogenetic analysis of excised ...
文摘A molecular biology method, fluorescent in situ hybridization(FISH), in which the pre-treatment was improved in allusion to the media of the constructed wetlands(CW), e.g. the soil and the grit, was used to investigate the vertical distribution characteristics of ammonia-oxidizing bacteria(AOB) quantity and the relation with oxidation-reduction potential(ORP) in the Typha latifolia constructed wetlands under three different Ioadings in summer from May to September. Results showed that the quantity of the AOB decreased in the Typha latifolia CW with the increase of vertical depth. However, the AOB quantity was 2-4 times the quantity of the control in the root area. Additionally, ORP in the rhizosphere was found to be higher than other areas, which showed that Typha latifolia CW was in an aerobic state in summer when using simulated non-point sewage at the rural area of Taihu Lake in China and small town combined sewage.
基金Supported by the Special Scientific Fund for Non-profit Environmental Industry(2010467038)~~
文摘ObjectiveThe aim was to understand the effects of transgenic DREB soybean on the ammonia-oxidizing bacteria. MethodThe diversity of the cto gene in pot-planted transgenic soybean and near-isogenic non-transgenic soybean under normal water condition and drought stress was analyzed by PCR-DGGE and sequence analysis. ResultRhizosphere community diversity of ammonia-oxidizing bacteria showed no difference between the treatments of transgenic soybean and its non-transgenic isolines, moreover transgenic soybean under normal water condition and drought stress improved the diversity of the ammonia-oxidizing bacteria in the harvest time. The phylogenetic analysis revealed that all the sequences of excised DGGE bands were closely related to members of the genus Nitrosovibrio and Nitrosospira of the β-subclass Proteobacteria. ConclusionTransgenic DREB soybean has no adverse impact on soil ammonia-oxidizing bacteria.
基金supported by the Key Projects in National Science & Technology Pillar Program during the Eleventh Five-Year Plan Period (No.2006BAC19B01-02)the Mega-projects of Science Research for Water (No.2008ZX07313-3)the Program of Introducing Talents of Discipline to Universities
文摘We investigated the communities of ammonia-oxidizing bacteria (AOB) in activated sludge collected from eight wastewater treatment systems using polymerase chain reaction (PCR) followed by terminal restriction fragment length polymorphism (T-RFLP), cloning, and sequencing of the α-subunit of the ammonia monooxygenase gene (amoA). The T-RFLP fingerprint analyses showed that different wastewater treatment systems harbored distinct AOB communities. However, there was no remarkable difference among the AOB T- RFLP profiles from different parts of the same system. The T-RFLP fingerprints showed that a full-scale wastewater treatment plant (WWTP) contained a larger number of dominant AOB species than a pilot-scale reactor. The source of influent affected the AOB community, and the WWTPs treating domestic wastewater contained a higher AOB diversity than those receiving mixed domestic and industrial wastewater. However, the AOB community structure was little affected by the treatment process in this study. Phylogenetic analysis of the cloned amoA genes clearly indicated that all the dominant AOB in the systems was closely related to Nitrosomonas spp. not to Nitrosospira spp. Members of the Nitrosomonas oligotropha and Nitrosomonas communis clusters were found in all samples, while members of Nitrosomonas europaea cluster occurred in some systems.
基金The National Fundamental Project of China under grant No.2006CB400602
文摘The spatial distribution of ammonia-oxidizing Betaproteobacteria (βAOB) was investigated by FISH (fluorescence in situ hybridization) and DGGE (denaturing gradient get electrophoresis) techniques in the sediment off the Changjiang River Estuary. Sediment samples were collected from eight stations in June before the formation of hypoxia zone in 2006. The abundance of βAOB ranged from 1.87× 10^5 to 3.53×10^5 cells/g of sediment. βAOB abundance did not present a negative correlation with salinity, whereas salinity was implicated as the primary factor affecting nitrification rates. The DGGE profiles of PCR-amplified amoA gene fragments revealed that the βAOB community structure of sample S2 separated from other samples at the level of 40% similarity. The variations in composition ofβAOB were significantly correlated with the salinity, temperature, absorption ability of sediments and TOC. The statistical analysis indicates that theβAOB abundance was a main factor to influence nitrification rates with an influence ratio of 87.7% at the level of 40% biodiversity similarity. Considering the good correlation between βAOB abundance and nitrification estimates, the abundance and diversity of βAOB community could be expected as an indirect index of nitrification activity at the study sea area in summer.
基金This research was supported by the National Key R&D Program of China(No.2017YFD0701700)the National Natural Science Foundation of China(Nos.31472312 and 31672673).
文摘We investigated the changes in communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira during the operation of a pufferfish Takifugu rubripes recirculating aquaculture system by using high-throughput DNA sequencing.Differences in bacterial communities were observed at days 1-32,47-62 and 78-93 of biofilm development by using 16S rRNA gene pyrosequencing.The relative abundance of Proteobacteria(Gammaproteobacteria)increased,while that of Bacteroidetes(Flavobacteria)decreased.The proportions of Nitrosomonas and Nitrospina ranged from 0.02%to 0.30%and from 0.02%to 0.83%,respectively.Ammonia monooxygenase gene pyrosequencing revealed that the top three operational taxonomic units were related to Nitrosomonas aestuarii(17.5%-61.1%),uncultured beta proteobacterium clone B67S-54(1.9%-45.2%),and uncultured bacterium clone AZPa8(3.6%-24.7%).Nitrite oxidoreductase gene pyrosequencing revealed that the relative abundance of the dominant strain Nitrospira sp.Ecomares 2.1 increased,but that of the abundant species Nitrospira marina decreased.Our results demonstrated that the communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira were changing during the operation of the pufferfish recirculating aquaculture system.
基金The National Natural Science Foundation of China(No.51479034,50909019)the Fundamental Research Funds for the Central Universities
文摘In order to explore the nitrogen removal process in constructed wetlands(CW s),the moisture,ammonia nitrogen(NH4+-N),nitrate nitrogen(NO3"-N)and nitrification intensity in three wetland plant rhizosphere soils(Acorns calamus,Typha orientalis,Iris pseudacorus)were investigated at a relatively normal temperature range of15to25The relative abundance of ammonia-oxidizing bacteria(AOB)and ammonia-oxidizing archaea(AOA)were also achieved using fluorescence in situ hybridization(FISH).It is found that T.orientalis achieves the highest nitrification intensity of2.03m g(h?kg)while the second is I.pseudacorrs(1.74m g/(h?kg)),and followed by A.calamus(1.65m g/(h?kg))throughout the experiment.FISH reveals that the abundance of bacteria(1010g_1wet soil)is higher than that of archaea(109g_1wet soil),and AOBare the dominant bacteria in the ammonia oxidation process.The abundance of AOB in te rhizosphere soils from high to low T.orientalis(1.88x1010g"1),I pseudacorus(1.23x1010g1),A.calamus(5.07x109g"1)while the abundance of AOA from high to low ae I.pseudacorus(4.00x109g1),A.calamus(3.52x109g"1),T.orientalis(3.48x109g"1).The study provides valuable evidence of plant selection for nitrogen removal in CWs.
基金the National Natural Science Foundation of China(31101504 and 31171917)the Postdoctoral Science Foundation of China(2011M500575)+1 种基金the China Agricultural Research System(CARS-28)the Shenyang Municipal Science and Technology Research Projects,China(F12-109-3-00)for their financial support
文摘In this study, we investigated the potential nitrification and community structure of soil-based ammonia-oxidizing bacteria (AOB) in apple orchard soil during different growth periods and explored the effects of environmental factors on nitrification activity and AOB community composition in the soil of a Hanfu apple orchard, using a culture-dependent technique and denaturing gradient gel electrophoresis (DGGE). We observed that nitrification activity and AOB abundance were the highest in November, lower in May, and the lowest in July. The results of statistical analysis indicated that total nitrogen (N) content, NH4+-N content, NO3-N content, and pH showed significant correlations with AOB abundance and nitrification activity in soil. The Shannon-Winner diversity, as well as species richness and evenness indices (determined by PCR-DGGE banding patterns) in soil samples were the highest in September, but the lowest in July, when compared to additional sampled dates. The DGGE fingerprints of soil-based 16S rRNA genes in November were apparently distinct from those observed in May, July, and September, possessing the lowest species richness indices and the highest dominance indices among all four growth periods. Fourteen DGGE bands were excised for sequencing. The resulting analysis indicated that all AOB communities belonged to the 13-Proteobacteria phylum, with the dominant AOB showing high similarity to the Nitrosospira genus. Therefore, soil-based environmental factors, such as pH variation and content of NHa+-N and NO3--N, can substantially influence the abundance of AOB communities in soil, and play a critical role in soil-based nitrification kinetics.
基金The National Key R&D Program of China under contract No.2022YFE0209300the National Natural Science Foundation of China under contract No.42006152+1 种基金the Zhejiang Provincial Natural Science Foundation of China under contract No.LQ21D060005the 111 Project under contract No.BP0820020.
文摘Subterranean estuaries(STEs)are characterized by the mixing of terrestrial fresh groundwater and seawater in coastal aquifers.Although microorganisms are important components of coastal groundwater ecosystems and play critical roles in biogeochemical transformations in STEs,limited information is available about how their community dynamics interact with hydrological,geochemical and environmental characteristics in STEs.Here,we studied bacterial and archaeal diversities and distributions with 16S rRNA-based Illumina MiSeq sequencing technology between surface water and groundwater in a karstic STE.Principal-coordinate analysis found that the bacterial and archaeal communities in the areas where algal blooms occurred were significantly separated from those in other stations without algal bloom occurrence.Canonical correspondence analysis showed that nutrients and salinity can explain the patterns of bacterial and archaeal community dynamics.The results suggest that hydrological,geochemical and environmental characteristics between surface water and groundwater likely control the bacterial and archaeal diversities and distributions in STEs.Furthermore,we found that some key species can utilize terrestrial pollutants such as nitrate and ammonia in STEs,indicating that these species(e.g.,Nitrosopumilus maritimus,Limnohabitans parvus and Simplicispira limi)may be excellent candidates for in situ degradation/remediation of coastal groundwater contaminations concerned with the nitrate and ammonia.Overall,this study reveals the coupling relationship between the microbial communities and hydrochemical environments in STEs,and provides a perspective of in situ degradation/remediation for coastal groundwater quality management.
基金the National Natural Science Foundation of China(40801097)the Natural Science Foundation of Fujian Province,China(2012J01107)
文摘Ammonia oxidation, the first and rate-limiting step of nitrification, is carried out by both ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA). However, the relative importance of AOB and AOA to nitrification in terrestrial ecosystems is not well understood. The aim of this study was to investigate the effect of the nitrogen input amount on abundance and community composition of AOB and AOA in red paddy soil. Soil samples of 10-20 cm (root layer soil) and 0-5 cm (surface soil) depths were taken from a red paddy. Rice in the paddy was fertilized with different rates of N as urea of N1 (75 kg N ha" yr-1), N2 (150 kg N ha~ yrl), N3 (225 kg N ha1 yrl) and CK (without fertilizers) in 2009, 2010 and 2011. Abundance and community composition of ammonia oxidizers was analyzed by real-time PCR and denaturing gradient gel electrophoresis (DGGE) based on amoA (the unit A of ammonia monooxygenase) gene. Archaeal amoA copies in N3 and N2 were significantly (P〈0.05) higher than those in CK and N1 in root layer soil or in surface soil under tillering and heading stages of rice, while the enhancement in bacterial amoA gene copies with increasing of N fertilizer rates only took on in root layer soil. N availability and soil NO3--N content increased but soil NH4+-N content didn't change with increasing of N fertilizer rates. Otherwise, the copy numbers of archaeal amoA gene were higher (P〈0.05) than those of bacterial amoA gene in root lary soil or in surface soil. Redundancy discriminate analysis based on DGGE bands showed that there were no obvious differs in composition of AOA or AOB communities in the field among different N fertilizer rates. Results of this study suggested that the abundance of ammonia-oxidizers had active response to N fertilizer rates and the response of AOA was more obvious than that of AOB. Similarity in the community composition of AOA or AOB among different N fertilizer rates indicate that the community composition of ammonia-oxidizers was relatively stable in the paddy soil at least in short term for three years.
基金This study was financially supported by the General Research Fund of the Research Grants Council,the Hong Kong Special Administrative Region,China(HKU 702612P and HKU 703010P).
文摘Ammonia-oxidizing archaea(AOA),ammonia-oxidizing bacteria(AOB),and anaerobic ammonia-oxidation(anammox)bacteria are very important contributors to nitrogen cycling in natural environments.Functional gene abundances of these microbes were believed to be well relevant to N-cycling in groundwater systems,especially in the Pearl River Delta(PRD)groundwater with unique high intrinsic ammonia concentrations.In this research,20 sediment samples from two in the PRD were collected for porewater chemistry analysis and quantification of N-cycling related genes,including archaeal and bacterial amoA gene and anammox 16S ribosomal Ribonucleic Acid(rRNA)gene.Quantitative Polymerase Chain Reaction(qPCR)results showed that gene abundances of AOA,AOB,and anammox bacteria ranged from 3.13×10^(5)to 3.21×10^(7),1.83×10^(4)to 2.74×10^(6),and 9.27×10^(4)to 8.96×10^(6)copies/g in the sediment of the groundwater system,respectively.Anammox bacteria and AOA dominated in aquitards and aquifers,respectively,meanwhile,the aquitard-aquifer interfaces were demonstrated as ammonium-oxidizing hotspots in the aspect of gene numbers.Gene abundances of nitrifiers were analyzed with geochemistry profiles.Correlations between gene numbers and environmental variables indicated that the gene abundances were impacted by hydrogeological conditions,and microbial-derived ammonium loss was dominated by AOA in the northwest PRD and by anammox bacteria in the southeast PRD.
基金supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant (RGPIN/227565-2013) to K.A.G
文摘The abundance of ammonia-oxidizing bacteria and archaea and their amo A genes from the aerobic activated sludge tanks,recycled sludge and anaerobic digesters of a full-scale wastewater treatment plant(WWTP)was determined.Polymerase chain reaction and denaturing gradient gel electrophoresis were used to generate diversity profiles,which showed that each population had a consistent profile although the abundance of individual members varied.In the aerobic tanks,the ammonia-oxidizing bacterial(AOB)population was more than 350 times more abundant than the ammonia-oxidizing archaeal(AOA)population,however in the digesters,the AOA population was more than 10 times more abundant.Measuring the activity of the amo A gene expression of the two populations using RT-PCR also showed that the AOA amo A gene was more active in the digesters than in the activated sludge tanks.Using batch reactors and dd PCR,amo A activity could be measured and it was found that when the AOB amo A activity was inhibited in the anoxic reactors,the expression of the AOA amo A gene increased fourfold.This suggests that these two populations may have a cooperative relationship for the oxidation of ammonia.
基金supported by a grant from the Tunisian Higher Education and Scientific Research Ministry
文摘Autotrophic ammonia-oxidizing bacteria (AOB) have been widely studied in constructed wetlands systems, while mixotrophic AOB have been less thoroughly examined. Heterotrophic bacteria were isolated from wastewater and rhizospheres of macrophytes of constructed wetlands, and then cultivated in a mixotrophic medium containing ammonium and acetic acid. A molecular characterization was accomplished using ITS-PCR amplification, and phylogenetic analysis based on 16S rRNA gene Sequences. Results showed the presence of 35 bacteria, among 400 initially heterotrophic isolates, that were able to remove ammonia. These 35 isolates were classified into 10 genetically different groups based on ITS pattern. Then, a collection of 10 isolates were selected because of their relatively high ammonia removal efficiencies (ARE≥ 80%) and their phylogenetic diversity. In conditions of mixotrophy, these strains were shown to be able to grow (increase of optical density OD660 during incubation with assimilation of nitrogen into cellular biomass) and to oxidize ammonia (important ammonia oxidation efficiencies, AOE between 79% and 87%). Among these facultative mixotrophic AOB, four isolates were genetically related to Firmicutes (Bacillus and Exiguobacterium), three isolates were affiliated to Actinobacteria (Arthrobacter) and three other isolates were associated with Proteobacteria (Pseudomonas, Ochrobactrum and Bordetella).
基金supported by grants from the National Natural Science Foundation of China(No.41701278)Shanghai Committee of Science and Technology(No.19DZ1204302)the Fundamental Research Funds for the Central Universities(No.2019MS041)。
文摘Ammonia-oxidizing bacteria(AOB)and archaea(AOA)are two microbial groups mediating nitrification,yet little is presently known about their abundances and community structures at the transcriptional level in wastewater treatment systems(WWTSs).This is a significant issue,as the numerical abundance of AOA or AOB at the gene level may not necessarily represent their functional role in ammonia oxidation.Using amo A genes as molecular markers,this study investigated the transcriptional abundance and community structure of active AOA and AOB in 14 WWTSs.Quantitative PCR results indicated that the transcriptional abundances of AOB amo A(averaged:1.6×10^(8)copies g^(-1)dry sludge)were higher than those of AOA(averaged:3.4×10^(7)copies g^(-1)dry sludge)in all WWTSs despite several higher abundances of AOA amo A at the gene level.Moreover,phylogenetic analysis demonstrated that Nitrosomonas europaea and unknown clusters accounted for 37.66%and 49.96%of the total AOB amo A transcripts,respectively,suggesting their dominant role in driving ammonia oxidation.Meanwhile,AOA amo A transcripts were only successfully retrieved from 3 samples,and the Nitrosospaera sister cluster dominated,accounting for 83.46%.Finally,the substrate utilization kinetics of different AOA and AOB species might play a fundamental role in shaping their niche differentiation,community composition,and functional activity.This study provides a basis for evaluating the relative contributions of ammonia-oxidizing microorganisms(AOMs)to nitrogen conversions in WWTSs.