Ammonia oxidizing (AOB) and denitrifying bacteria (DNB) play an important role in soil nitrogen transformation in natural and agricultural ecosystems. Effects of long-term fertilization on abundance and community ...Ammonia oxidizing (AOB) and denitrifying bacteria (DNB) play an important role in soil nitrogen transformation in natural and agricultural ecosystems. Effects of long-term fertilization on abundance and community composition of AOB and DNB were studied with targeting ammonia monooxygenase (amoA) and nitrite reductase (nirK) genes using polymerase chain reaction- denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR, respectively. A field trial with different fertilization treatments in a rice paddy from Tai Lake region, centre East China was used in this study, including no fertilizer application (NF), balanced chemical fertilizers (CF), combined organic/inorganic fertilizer of balanced chemical fertilizers plus pig manure (CFM), and plus rice straw return (CFS). The abundances and riehnesses of amoA and nirK were increased in CF, CFM and CFS compared to NF. Principle component analysis of DGGE profiles showed significant difference in nirK and amoA genes composition between organic amended (CFS and CFM) and the non-organic amended (CF and NF) plots. Number of amoA copies was significantly positively correlated with normalized soil nutrient richness (NSNR) of soil organic carbon (SOC) and total nitrogen (T-N), and that of nirK copies was with NSNR of SOC, T-N plus total phosphorus. Moreover, nitrification potential showed a positive correlation with SOC content, while a significantly lower denitrification potential was found under CFM compared to under CFS. Therefore, SOC accumulation accompanied with soil nutrient richness under long-term balanced and organic/inorganic combined fertilization promoted abundance and diversity of AOB and DNB in the rice paddy.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
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
This study examines the impacts of short-term(6 months) fertilization on the community structure and abundance of ammonia-oxidizing betaproteobacteria(β-AOB) and the potential nitrification rate in sediment colonized...This study examines the impacts of short-term(6 months) fertilization on the community structure and abundance of ammonia-oxidizing betaproteobacteria(β-AOB) and the potential nitrification rate in sediment colonized by S uaeda heteroptera in a saltmarsh located in Shuangtai estuary, China. The sediment samples were collected from plots treated with different amounts of an N fertilizer(urea supplied at 0.1, 0.2, 0.4, and 0.8 g/kg(nitrogen content in dry sediment)), and with different forms of N fertilizers(urea,(NH4) 2 SO 4, and NH_4NO_3, each supplied at 0.2 g/kg). The fertilizers were applied 1–4 times during the plant-growing season in May, July, August and September of 2013. Untreated plots were included as a control. As revealed in denaturing gradient gel electrophoresis of the 16 S r RNA gene, the β-AOB community responded to both the amount and form of N. Real-time quantitative PCR indicated that both abundance and potential nitrification rate of β-AOB increased after N addition, regardless of concentration and form(except NH_4NO_3). These results provide evidence that short-term N application influences the sediment β-AOB community, β-AOB abundance and potential nitrification rate in a saltmarsh ecosystem.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
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
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
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
基金supported by the National Natural Science Foundation of China(40830528 and 40710019002)
文摘Ammonia oxidizing (AOB) and denitrifying bacteria (DNB) play an important role in soil nitrogen transformation in natural and agricultural ecosystems. Effects of long-term fertilization on abundance and community composition of AOB and DNB were studied with targeting ammonia monooxygenase (amoA) and nitrite reductase (nirK) genes using polymerase chain reaction- denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR, respectively. A field trial with different fertilization treatments in a rice paddy from Tai Lake region, centre East China was used in this study, including no fertilizer application (NF), balanced chemical fertilizers (CF), combined organic/inorganic fertilizer of balanced chemical fertilizers plus pig manure (CFM), and plus rice straw return (CFS). The abundances and riehnesses of amoA and nirK were increased in CF, CFM and CFS compared to NF. Principle component analysis of DGGE profiles showed significant difference in nirK and amoA genes composition between organic amended (CFS and CFM) and the non-organic amended (CF and NF) plots. Number of amoA copies was significantly positively correlated with normalized soil nutrient richness (NSNR) of soil organic carbon (SOC) and total nitrogen (T-N), and that of nirK copies was with NSNR of SOC, T-N plus total phosphorus. Moreover, nitrification potential showed a positive correlation with SOC content, while a significantly lower denitrification potential was found under CFM compared to under CFS. Therefore, SOC accumulation accompanied with soil nutrient richness under long-term balanced and organic/inorganic combined fertilization promoted abundance and diversity of AOB and DNB in the rice paddy.
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 ...
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Supported by the National Natural Science Foundation of China(No.41171389)the Public Science and Technology Research Funds Projects of Ocean(No.201305043)the Program for Liaoning Excellent Talents in University(No.LR2013035)
文摘This study examines the impacts of short-term(6 months) fertilization on the community structure and abundance of ammonia-oxidizing betaproteobacteria(β-AOB) and the potential nitrification rate in sediment colonized by S uaeda heteroptera in a saltmarsh located in Shuangtai estuary, China. The sediment samples were collected from plots treated with different amounts of an N fertilizer(urea supplied at 0.1, 0.2, 0.4, and 0.8 g/kg(nitrogen content in dry sediment)), and with different forms of N fertilizers(urea,(NH4) 2 SO 4, and NH_4NO_3, each supplied at 0.2 g/kg). The fertilizers were applied 1–4 times during the plant-growing season in May, July, August and September of 2013. Untreated plots were included as a control. As revealed in denaturing gradient gel electrophoresis of the 16 S r RNA gene, the β-AOB community responded to both the amount and form of N. Real-time quantitative PCR indicated that both abundance and potential nitrification rate of β-AOB increased after N addition, regardless of concentration and form(except NH_4NO_3). These results provide evidence that short-term N application influences the sediment β-AOB community, β-AOB abundance and potential nitrification rate in a saltmarsh ecosystem.
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.