Temporal characteristics of algae-denitrifying bacteria co-occurrence patterns and denitrifier assembly in epiphytic biofilms on submerged macrophytes in Caohai Lake,SW China

Denitrifying bacteria in epiphytic biofilms play a crucial role in nitrogen cycle in aquatic habitats.However,little is known about the connection between algae and denitrifying bacteria and their assembly processes in epiphytic biofilms.Epiphytic biofilms were collected from submerged macrophytes(Patamogeton lucens and Najas marina L.)in the Caohai Lake,Guizhou,SW China,from July to November 2020 to:(1)investigate the impact of abiotic and biotic variables on denitrifying bacterial communities;(2)investigate the temporal variation of the algae-denitrifying bacteria co-occurrence networks;and(3)determine the contribution of deterministic and stochastic processes to the formation of denitrifying bacterial communities.Abiotic and biotic factors influenced the variation in the denitrifying bacterial community,as shown in the Mantel test.The co-occurrence network analysis unveiled intricate interactions among algae to denitrifying bacteria.Denitrifying bacterial community co-occurrence network complexity(larger average degrees representing stronger network complexity)increased continuously from July to September and decreased in October before increasing in November.The co-occurrence network complexity of the algae and nirS-encoding denitrifying bacteria tended to increase from July to November.The co-occurrence network complexity of the algal and denitrifying bacterial communities was modified by ammonia nitrogen(NH_(4)^(+)-N)and total phosphorus(TP),pH,and water temperature(WT),according to the ordinary least-squares(OLS)model.The modified stochasticity ratio(MST)results reveal that deterministic selection dominated the assembly of denitrifying bacterial communities.The influence of environmental variables to denitrifying bacterial communities,as well as characteristics of algal-bacterial co-occurrence networks and the assembly process of denitrifying bacterial communities,were discovered in epiphytic biofilms in this study.The findings could aid in the appropriate understanding and use of epiphytic biofilms denitrification function,as well as the enhancement of water quality.

Responses of N_2O reductase gene(nosZ)-denitrifier communities to long-term fertilization follow a depth pattern in calcareous purplish paddy soil

The effect of long-term fertilization on soil denitrifying communities was analysed by measuring the abundance and diversity of the nitrous oxide （N2O） reductase gene, nosZ. Soil samples were collected from plots of a long-term fertilization experiment established in 1982 in Suining City, China. The fertilizer treatments were no fertilizer （CK）, three chemical fertilizer （CF） treatments （N, NP, NPK）, manure （M） alone, and manure with chemical fertilizers （NM, NPM, NPKM）. The abundance and diversity of the denitrifying bacteria were assessed by real-time quantitative PCR, terminal restriction fragment length polymorphism （T-RFLP）, and cloning and sequencing of nosZ genes. The diversity and abundance of nosZ-denitrifiers was higher in soil amended with manure and chemical fertilizers （CFM） than in soil amended with CF alone, and the highest in topsoil （0-20 cm）. The nosZ-denitrifier community composition was more complex in CFM soil than in CF soil： Specific species were detected only in the CFM soil. The abundance of nosZ-denitrifier in the NPKM treatment was approximately two times higher than that in the CK, N, and NPK treatments. Most of the cloned nosZ sequences were closely related to nosZ sequences from Bradyrhizobiaceae and Rhodospirillaceae in Alphaproteobacteria. Of the measured abiotic factors, soil organic matter correlated significantly with the abundance （P〈0.01）; available phosphorus correlated significantly with the topsoil community composition （P〈0.01）, whereas soil organic matter correlated significantly with the subsoil （20-90 cm） community composition （P〈0.01）. This study demonstrated that long-term CFM fertilization affected both the abundance and composition of the nosZ-denitrifier community.

Effects of organic carbon consumption on denitrifier community composition and diversity along dissolved oxygen vertical profiles in lake sediment surface

At present,the understanding of the dynamics of denitrifiers at different dissolved oxygen(DO)layers under organic carbon consumption within the surface sediments remains inadequate.In this study,high-throughput sequencing and quantitative PCR targeting nirS gene were used to analyze the denitrifier abundance dynamics,community composition,and structure for aerobic(DO 0.5-6.9 mg/L),hypoxic-anoxic(DO 0-0.5 mg/L),and anoxic(DO 0 mg/L)layers in surface sediments under organic carbon consumption.Based on the analysis of nirS gene abundance,the values of denitrifying bacteria decreased with organic carbon consumption at different DO layers.When the bacterial species abundance at the genus level were compared between the high-carbon and low-carbon sediments,there was significant increase in 6 out of 36,7 out of 36 and 6 out of 36 genera respectively for the aerobic,hypoxic-anoxic and anoxic layers.On the other hand,14 out of 36,9 out of 36 and 15 out of 36 genera showed significant decrease in bacterial species abundance respectively for the aerobic,hypoxic-anoxic and anoxic layers.Additionally,14 out of 36,20 out of 36,and 15 out of 36 genera had no change in bacterial species abundance respectively for the aerobic,hypoxic-anoxic,and anoxic layers.This indicates that the carbon utilization ability of different denitrifiers on each DO layers was generally different from each other.Diversity of denitrifying bacteria also presented significant differences in different DO layers between the high-and low-carbon content sediment layers.Moreover,under the high-carbon and low-carbon content,the abundance of nirS gene showed a high peak within the hypoxic-anoxic regions,suggesting that this region might be the main distribution area for the denitrifying bacteria within the surface sediments.Furthermore,community of unique denitrifiers occurred in different DO layers and the adaptive changes of the denitrifier community followed the organic carbon consumption.

Isolation of aerobic denitrifier and characteristics of enhanced nitrogen removal in concentric-circles reactor with diversion wall

Three strains of aerobic denitrifiers,named as AT3,AT6 and AT7,were isolated from concentric-circles reactor with diversion wall possessing simultaneous nitrification and denitrification (SND) effect of 69%. The three strains are all gram-positive and rod-shaped,and their colonial colors are pale yellow,milk white and pink,respectively. Combined with 16SrDNA sequence homology comparison and biochemical tests,AT3 and AT7 were identified to belong to Rhodococcus,and AT6 to Gordonia. These bacterial strains could grow well in the medium with potassium nitrate as nitrogen source and sodium citrate as carbon source. Based on the enhanced nitrogen removal experiments of selected bacteria mixture for activated sludge,the inoculum amount of 5% was supposed to be proper. The mixed biomass suspension of selected strains with PVA immobilization was put into the concentric-circles reactor in order to study the characteristics of enhanced nitrogen removal after amplifying cultivation with inoculated amount of 5%. The experimental results show that the average removal efficiencies of ammonia nitrogen (NH3-N) and total nitrogen (TN) in the reactor enhanced with aerobic denitrifying bacteria using PVA are 92.18% and 79.14% respectively,increasing by 5.29% and 7.83% respectively compared with removal effects of control group without strains enhancement.

Study on separation and induction of high efficient denitrifier

A kind of denitrifier HY-1 initially obtained from activated sludge was domesticated and inducted with UV and illumination to a new species bacterium HY-2 that has high bioactivity at low temperature. HY-2 was most active at 13 ℃. Nitrate and CODcr removal efficiency was investigated under different temperature and C/N ratio.

Chronic effects of different fertilization regimes on nirS-type denitrifier communities across the black soil region of Northeast China

Denitrification is one of the major processes causing nitrogen loss from arable soils.This study aimed to investigate the responses of nir S-type denitrifier communities to different chronic fertilization regimes across the black soil region of Northeast China.Soil samples were collected from sites located in the north(NB),middle(MB),and south(SB)of the black soil region of Northeast China,each with four chronic fertilization regimes:no fertilizer(No F),chemical fertilizer(CF),manure(M),and chemical fertilizer plus manure(CFM).Methods of quantitative polymerase chain reaction(q PCR)and Illumina Mi Seq sequencing were applied to assess the abundance and composition of denitrifier communities by targeting the nir S gene.The results showed that the M and CFM regimes significantly increased the abundances of nir S-type denitrifiers compared with No F at the three locations.The majority of nir S sequences were grouped as unclassified denitrifiers,and the different fertilizers induced little variation in the relative abundance of known nir S-type denitrifier taxa.Over 90%of the sequences were shared among the four fertilization regimes at each location,but none of the abundant operational taxonomic units(OTUs)were shared among the three locations.Principal coordinate analysis(PCo A)revealed that the communities of nir S-type denitrifier were separated into three groups that corresponded with their locations.Although similar fertilization regimes did not induce consistent changes in the nir S-type denitrifier communities,soil p H and NO-3-N content simultaneously and significantly influenced the structure of nir S-type denitrifier communities at the three locations.Our results highlight that geographical separation rather than chronic fertilization was the dominant factor determining the nir S-type denitrifier community structures,and similar chronic fertilization regimes did not induce consistent shifts of nir S-type denitrifier communities in the black soils.

Factors influencing the accuracy of the denitrifier method for determining the oxygen isotopic composition of nitrate

The denitrifier method is widely used as a novel pretreatment method for the determination of nitrogen and oxygen isotope ratios as it can provide quantitative and high-sensitivity measurements. Nevertheless, the method is limited by relatively low measurement accuracy for δ18 O. In this study, we analyzed the factors influencing the accuracy of δ18 O determination, and then systematically investigated the effects of dissolved oxygen concentrations and nitrate sample sizes on estimates of the δ15 N and δ18 O of nitrate reference materials. The δ18 O contraction ratio was used to represent the relationship between the measured difference and true difference between two reference materials. We obtained the following main results:(1) a gas-liquid ratio of 3:10(v/v) in ordinary triangular flasks and a shaking speed of 120 r/min produced an optimal range(1.9 to 2.6 mg/L) in the concentration of dissolved oxygen for accurately determining δ18 O, and(2) the δ18 O contraction ratio decreased as nitrate sample size decreased within a certain range(1.0 to 0.1 μmol). Our results suggested that δ18 O contraction is influenced mainly by dissolved oxygen concentrations in pure culture, and provided a model for improving the accuracy of oxygen isotope analysis.

Study on the Microorganisms Related with Nitrogen Cycle in Sediments of Hongfeng Lake

Hongfeng Lake,a key drinking-water source located in Karst areas,was selected to analyze the microorganisms associated with nitrogen cycle.Dilution plate method and most probable number method were used to determine the distribution condition of ammonifier,ammonium-oxidizing bacteria,nitrite-oxidizing bacteria and denitrifier in surface sediment in eight sites.The results showed that the amount of ammonifier,ammonium-oxidizing bacteria,nitrite-oxidizing bacteria and denitrifier in eight sites were 104-106 CFU/g,105-107 MPN/g,105-108 MPN/g and 105-109 MPN/g,respectively.The sampling site with the largest amount of denitrifier was Daposhang,and the other sampling sites had no significant difference;the sampling site of Huyudong bridge had the largest amount of ammonium-oxidizing bacteria;the amount of nitrite-oxidizing bacteria in Xinzhuang was higher than that of other sampling sites;the sampling site of Daposhang had the larger amount of denitrifier.

缢蛏滩涂养殖环境的细菌群落组成及分析

In this study,the bacteria from the mud in tidal-flat Sinonovacula constricta aquaculture area were isolated each month from March to December,2002,and the temporal and spatial distribution of heterotrophic bacteria,ammonifying bacteria,denitrifying bacteria,and sulphate reducing bacteria were analyzed.The results showed that all the 515 isolated bacteria mainly belonged to 1 family and 13 genera.The bacterial flora in different layers of the mud was almost consistent,while the composition was different.The predominant genera were Clostridium,Bacillus,Corynebacterium,Photobacterium,and some Enterobacteriaceae.The number of heterotrophic bacteria in the surface layer and the bottom fluctuated in 7.6×103 cfu·g-1～2.0×105 and 1.6×103～1.0×105 cfu·g-1,ammonifying bacteria fluctuated in 1.5×106～9.0×107 and 9.0×105～1.0×107 cfu·g-1,denitrifying bacteria fluctuated in 9.0×103～4.0×106 and 5.0×102～1.9×106 cfu·g-1,and sulphate reducing bacteria fluctuated in 5.0×104～5.0×106 and 1.9×104～2.0×106 cfu·g-1,respectively.The detection rates of ammonifying bacteria,denitrifying bacteria and sulphate reducing bacteria in the mud were all 100%,and these bacteria increased significantly in the second half of the year,indicating that the environment of the Sinonovacula constricta aquaculture area was deteriorated due to the accumulation of NH3,nitrite and H2S,and it is important to regulate the breed capacity and redistribute the breeding environment.

Effects of long-term elevated CO_2 on N_2-fixing,denitrifying and nitrifying enzyme activities in forest soils under Pinus sylvestriformis in Changbai Mountain

A study was conducted to determine the effects of elevated CO2 on soil N process at Changbai Mountain in Jilin Province, northeastern China （42°24＂N, 128°06＂E, and 738 m elevation）. A randomized complete block design of ambient and elevated CO2 was established in an open-top chamber facility in the spring of 1999. Changpai Scotch pine （Pinus sylvestris var. sylvestriformis seeds were sowed in May, 1999 and CO2 fumigation treatments began after seeds germination. In each year, the exposure started at the end of April and stopped at the end of October. Soil samples were collected in June and August 2006 and in June 2007, and soil nitrifying, denitrifying and N2-fixing enzyme activities were measured. Results show that soil nitrifying enzyme activities （NEA） in the 5-10 cm soil layer were significantly increased at elevated CO2 by 30.3% in June 2006, by 30.9% in August 2006 and by 11.3% in June 2007. Soil denitrifying enzyme activities （DEA） were significantly decreased by elevated CO2 treatment in June 2006 （P 〈 0.012） and August 2006 （P 〈 0.005） samplings in our study; no significant difference was detected in June 2007, and no significant changes in N2-fixing enzyme activity were found. This study suggests that elevated CO2 can alter soil nitrifying enzyme and denitrifying enzyme activities.

污水处理反硝化除磷-诱导结晶磷回收工艺中除磷微生物特性

为探究反硝化除磷-诱导结晶磷回收工艺中缺氧池污泥释磷、吸磷以及微生物特征,利用荧光原位杂交(fluorescence in situ hybridization,FISH)技术、电子扫描显微镜(scanning electron microscope,SEM)观察了微生物的数量、分布和形态;通过批次试验考察了污泥在厌氧/好氧和厌氧/缺氧2种模式下的释磷和吸磷特征。结果表明:该双污泥系统缺氧池中聚磷菌占总细菌比例的69.7%,明显高于单污泥系统中富集的聚磷菌比例,污泥中的微生物多呈杆状;厌氧/好氧、厌氧/缺氧模式下单位污泥浓度(mixed liquor suspended solids,MLSS)总吸磷量(以PO43--P计)分别为22.84、18.60 mg/g,反硝化聚磷菌(denitrifying polyphosphate-accumulating organisms,DPAO)占聚磷菌(polyphosphate-accumulating organisms,PAO)的比例为81.44%,表明在长期的厌氧/缺氧运行条件下可以富集到以硝酸盐为电子受体的反硝化聚磷菌,同时还存在着仅以氧气为电子受体的聚磷菌;通过pH值和氧化还原电位(oxidation reduction potential,ORP)的实时监测可以快速地了解污水生物处理系统中各类反应的进程,对调控工艺参数有着重要的意义。综上所述,为保证污水生物处理工艺的正常稳定运行,将微生物分析与常规的化学参数分析结合起来考察将是未来发展的必然趋势。

Isolation of Denitrifying Bacteria in Sea Sediment and Simulated Experiment of Removing Nitrate-N from Seawater

The objective of this research was to isolate denitrifying bacteria from sea sediment and simulate the removal efficiency of nitrate-N by denitrifying bacteria from seawater. The result showed that the isolated denitrifying bacteria could effectively remove nitrate-N from seawater. About 90 % of nitrate-N was removed by denitrifying bacteria from seawater within a week in the simulated experiment I （the initial concentration of nitrate-N was 100 mg/L）. The removal efficiency of nitrate-N reached about 70 % within one day in the simulated experiment Ⅱ （initial concentration of nitrate-N was 1 mg/L）. The final removal efficiency was about 98 % and 85 % in the simulated experiments Ⅰ and Ⅱ, respectively. It was found that there was positive correlation between the concentration of nitrate-N and the number of denitrifying bacteria in seawater. Lots of denitrifying bacteria would disappear and the seawater would become transparent once the process of bioremediation was completed.

Microbial Characterization of Denitrifying Sulfide Removal Sludge Using High-Throughput Amplicon Sequencing Method

The denitrifying sulfide removal(DSR) process has recently been studied extensively from an engineering perspective. However, the importance of microbial communities of this process was generally underestimated. In this study, the microbial community structure of a lab-scale DSR reactor was characterized in order to provide a comprehensive insight into the key microbial groups in DSR system. Results from high-throughput sequencing analysis revealed that the fraction of autotrophic denitrifiers increased from 2.34 % to 10.93% and 44.51% in the DSR system when the influent Na Cl increased from 0 g/L, to 4 g/L and 30 g/L, respectively. On the contrary, the fraction of heterotrophic denitrifiers decreased from 61.74% to 39.57%, and 24.12%, respectively. Azoarcus and Thiobacillus were the main autotrophic denitrifiers, and Thauera was the main hetetrophic denitrifier during the whole process. This study could be useful for better understanding the interaction between autotrophs and heterotrophs in DSR system.

Characteristics of anoxic phosphors removal in sequence batch reactor

The characteristics of anaerobic phosphorus release and anoxic phosphorus uptake were investigated in sequencing batch reactors using denitrifying phosphorus removing bacteria （DPB） sludge. The lab-scale experiments were accomplished under conditions of various nitrite concentrations （5.5, 9.5, and 15 mg/L） and mixed liquor suspended solids （MLSS） （1844, 3231, and 6730 mg/L）. The results obtained confirmed that nitrite, MLSS, and pH were key factors, which had a significant impact on anaerobic phosphorus release and anoxic phosphorus uptake in the biological phosphorous removal process. The nitrites were able to successfully act as electron acceptors for phosphorous uptake at a limited concentration between 5.5 and 9.5 mg/L. The denitrification and dephosphorous were inhibited when the nitrite concentration reached 15 mg/L. This observation indicated that the nitrite would not inhibit phosphorus uptake before it exceeded a threshold concentration. It was assumed that an increase of MLSS concentration from 1844 mg/L to 6730 mg/L led to the increase of denitrification and anoxic P-uptake rate. On the contrary, the average P-uptake/N denitrifying reduced from 2.10 to 1.57 mg PO4^3--P/mg NO3^--N. Therefore, it could be concluded that increasing MLSS of the DEPHANOX system might shorten the reaction time of phosphorus release and anoxic phosphorus uptake. However, excessive MLSS might reduce the specific denitrifying rate. Meanwhile, a rapid pH increase occurred at the beginning of the anoxic conditions as a result of denitrification and anoxic phosphate uptake. Anaerobic P release rate increased with an increase in pH. Moreover, when pH exceeded a relatively high value of 8.0, the dissolved P concentration decreased in the liquid phase, because of chemical precipitation. This observation suggested that pH should be strictly controlled below 8.0 to avoid chemical precipitation if the biological denitrifying phosphorus removal capability is to be studied accurately.

Denitrifying phosphorus removal in a step-feed CAST with alternating anoxic-oxic operational strategy

A bench-scale cyclic activated sludge technology （CAST） was operated to study the biological phosphorus removal performance and a series of batch tests was carried out to demonstrate the accumulation of denitrifying polyphosphate-accumulating organisms （DNPAOs） in CAST system. Under all operating conditions, step-feed CAST with enough carbon sources in influent had the highest nitrogen and phosphorus removal efficiency as well as good sludge settling performance. The average removal rate of COD, NH4^＋-N, PO4^3--P and total nitrogen （TN） was 88.2%, 98.7%, 97.5% and 92.1%, respectively. The average sludge volume index （SVI） was 133 mL/g. The optimum anaerobic/aerobic/anoxic （AOA） conditions for the cultivation of DNPAOs could be achieved by alternating anoxic/oxic operational strategy, thus a significant denitrifying phosphorus removal occurred in step-feed CAST. The denitrification of NO^x--N completed quickly due to step-feed operation and enough carbon sources, which could enhance phosphorus release and further phosphorus uptake capability of the system. Batch tests also proved that polyphosphate-accumulating organisms （PAOs） in the step-feed process had strong denitrifying phosphorus removal capacity. Both nitrate and nitrite could be used as electron acceptors in denitrifying phosphorus removal. Low COD supply with step-feed operation strategy would favor DNPAOs accumulation.

Influence of pH on short-cut denitrifying phosphorus removal

Through a series of experiments using denitrifying phosphorus-accumulating sludge in sequencing batch reactors （SBRs）, the variations of the intracellular polymers during the anaerobic phosphorus release process at different pH values were compared, the probable reasons for different performances of phosphorus removal were examined, and system operations in a typical cycle were investigated. The results show that the phosphorus removal rate was positively correlated with pH values in a range of 6.5-8.5. When the pH value was 8.0, the anaerobic phosphorus release rate and anoxic phosphorus uptake rate of the activated sludge were 20.95 mg/（g, h） and 23.29 mg/（g, h）, respectively; the mass fraction of poly-13-hydroxybutyrate （PHB） increased to 62.87 mg/g under anaerobic conditions; the mass fraction of polyphosphate was 92.67 mg/g under anoxic conditions; and the effluent concentration of total phosphorus （TP） was 1.47 mg/L. With the increase of pH, the mass fraction of acetic acid and PHB also increased, and the absorption rate of acetic acid was equal to the disintegration rate of polyphosphate. When the pH value was above 8.0, biological phosphorus removal was achieved by chemical phosphorus precipitation, and the phosphorus removal rate decreased.

Removal of nitrogen and phosphorus in a combined A^2/O-BAF system with a short aerobic SRT

A bench-scale anaerobic/anoxic/aerobic process-biological aerated filter （A^2/O-BAF） combined system was carded out to treat wastewater with lower C/N and C/P ratios. The A^2/O process was operated in a short aerobic sludge retention time （SRT） for organic pollutants and phosphorus removal, and denitrification. The subsequent BAF process was mainly used for nitrification. The BAF effluent was partially returned to anoxic zone of the A^2/O process to provide electron acceptors for denitrification and anoxic P uptake. This unique system formed an environment for reproducing the denitdfying phosphate-accumulating organisms （DPAOs）. The ratio of DPAOs to phosphorus accumulating organisms （PAOs） could be maintained at 28% by optimizing the organic loads in the anaerobic zone and the nitrate loads into the anoxic zone in the A^2/O process. The aerobic phosphorus over-uptake and discharge of excess activated sludge was the main mechanism of phosphorus removal in the combined system. The aerobic SRT of the A^2/O process should meet the demands for the development of aerobic PAOs and the restraint on the nitrifiers growth, and the contact time in the aerobic zone of the A^2/O process should be longer than 30 min, which ensured efficient phosphorus removal in the combined system. The adequate BAF effluent return rates should be controlled with 1--4 mg/L nitrate nitrogen in the anoxic zone effluent of A^2/O process to achieve the optimal nitrogen and phosphorus removal efficiencies.

Preliminary investigation on the role of microorganisms in the production of phosphine

The relationships between the phosphine content and various microbial populations,activities of different enzymes were investigated firstly.The results indicated that the phosphine content of samples from various environments was positively related to total anaerobic microorganisms,organic phosphate compound-dissolving bacteria,denitrifying bacteria,and the activities of alkaline phosphatase and dehydrogenase,with correlation coefficients (R^2) up to 0.93,0.90,0.69,0.79,and 0.82,respectively.Results also sh...

Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria

To investigate the characteristics and metabolic mechanism of short-cut denitrifying phospho- rus-removing bacteria （SDPB） that are capable of enhanced biological phosphorus removal （EBPR） using nitrite as an electron acceptor, an aerobic/anoxic sequencing batch reactor was operated under three phases. An SDPB-strain YC was screened after the sludge enrichment and was identified by morphological, physiological, biochemical properties and 16S rDNA gene sequence analysis. Denitrifying phosphorus-removing experiments were conducted to study anaerobic and anoxic metabolic mechanisms by analyzing the changes of chemical oxygen demand （COD）, phosphate, nitrite, poly-fl-hydroxybutyrate （PHB）, and glycogen. The results show that strain YC is a non-fermentative SDPB similar to Paracoccus denitrificans. As a kind of non-fermentative bacteria, the energy of strain YC was mainly generated from phosphorus release （96.2%） under anaerobic conditions with 0.32 mg P per mg synthesized PHB. Under anoxic conditions, strain YC accumulated 0.45 mg P per mg degraded PHB, which produced most of energy for phosphate accumulation （91.3%） and a little for glycogen synthesis （8.7%）. This metabolic mechanism of strain YC is different from that of traditional phosphorus-accumulating organisms （PAOs）. It is also found that PHB, a kind of intracellular polymer, plays a very important role in denitrifying and accumulating phosphorus by supplying sufficient energy for phosphorous accumulation and carbon sources for denitrification. Therefore, monitoring AP/APHB and ANO2 -N/APHB is more necessary than monitoring AP/ACOD, ANO2 -N/ACOD, or AP / ANO2 -N.

Impact of Long-Term Fertilization on Community Structure of Ammonia Oxidizing and Denitrifying Bacteria Based on amoA and nirK Genes in a Rice Paddy from Tai Lake Region,China

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.