Potential Secretory Transporters and Biosynthetic Precursors of Biological Nitrification Inhibitor 1,9-Decanediol in Rice as Revealed by Transcriptome and Metabolome Analyses

Biological nitrification inhibitors(BNIs)are released from plant roots and inhibit the nitrification activity of microorganisms in soils,reducing NO_(3)^(‒)leaching and N2O emissions,and increasing nitrogenuse efficiency(NUE).Several recent studies have focused on the identification of new BNIs,yet little is known about the genetic loci that govern their biosynthesis and secretion.We applied a combined transcriptomic and metabolomic analysis to investigate possible biosynthetic pathways and transporters involved in the biosynthesis and release of BNI 1,9-decanediol(1,9-D),which was previously identified in rice root exudates.Our results linked four fatty acids,icosapentaenoic acid,linoleate,norlinolenic acid,and polyhydroxy-α,ω-divarboxylic acid,with 1,9-D biosynthesis and three transporter families,namely the ATP-binding cassette protein family,the multidrug and toxic compound extrusion family,and the major facilitator superfamily,with 1,9-D release from roots into the soil medium.Our finding provided candidates for further work on the genes implicated in the biosynthesis and secretion of 1,9-D and pinpoint genetic loci for crop breeding to improve NUE by enhancing 1,9-D secretion,with the potential to reduce NO_(3)^(‒)leaching and N2O emissions from agricultural soils.

Improving dryland maize productivity and water efficiency with heterotrophic ammonia-oxidizing bacteria via nitrification and cytokinin activity

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.

Alleviated photoinhibition on nitrification in the Indian Sector of the Southern Ocean

Nitrification,a central process in the marine nitrogen cycle,produces regenerated nitrate in the euphotic zone and emits N_(2)O,a potent greenhouse gas as a by-product.The regulatory mechanisms of nitrification in the Southern Ocean,which is a critical region for CO_(2)sequestration and radiative benefits,remain poorly understood.Here,we investigated the in situ and dark nitrification rates in the upper 500 m and conducted substrate kinetics experiments across the Indian Sector in the Cosmonaut and Cooperation seas in the late austral summer.Our findings indicate that light inhibition of nitrification decreases exponentially with depth,exhibiting a light threshold of 0.53%photosynthetically active radiation.A positive relationship between dark nitrification and apparent oxygen utilization suggests a dependence on substrate availability from primary production.Importantly,an increased NH_(4)^(+) supply can act as a buffer against photo-inhibitory damage.Globally,substrate affinity(α)increases with depth and transitions from light to dark,decreases with increasing ambient NH_(4)^(+)and exhibits a latitudinal distribution,reflecting substrate utilization strategies.We also reveal that upwelling in Circumpolar Deep Water(CDW)stimulates nitrification through the introduction of potentially higher iron and deep diverse nitrifying microorganisms with higherα.We conclude that although light is the primary limiting factor for nitrification in summer,coupling between substrate availability and CDW upwelling can overcome this limitation,thereby alleviating photoinhibition by up to 45%±5.3%.

Simultaneous nitrification and autotrophic denitrification in fluidized bed reactors using pyrite and elemental sulfur as electron donors

In this study, simultaneous nitrification and autotrophic denitrification (SNAD) with either elemental sulfur or pyrite were investigated in fluidized bed reactors in mesophilic conditions. The reactor performance was evaluated at different ammonium (12-40 mg/L of NH4+-N), nitrate (35-45 mg/L of NO3--N), and dissolved oxygen (DO) (0.1-1.5 mg/L) concentrations, with a hydraulic retention time of 12 h. The pyrite reactor supported the SNAD process with a maximum nitrogen removal efficiency of 139.5 mg/(L·d) when the DO concentration was in the range of 0.8-1.5 mg/L. This range, however, limited the denitrification efficiency of the reactor, which decreased from 90.0% ± 5.3% in phases II-V to 67.9% ± 7.2% in phases VI and VII. Sulfate precipitated as iron sulfate (FeSO4/Fe2(SO4)3) and sodium sulfate (Na2SO4) minerals during the experiment. The sulfur reactor did not respond well to nitrification with a low and unstable ammonium removal efficiency, while denitrification occurred with a nitrate removal efficiency of 97.8%. In the pyrite system, the nitrifying bacterium Nitrosomonas sp. was present, and its relative abundance increased from 0.1% to 1.1%, while the autotrophic denitrifying genera Terrimonas, Ferruginibacter, and Denitratimonas dominated the community. Thiobacillus, Sulfurovum, and Trichlorobacter were the most abundant genera in the sulfur reactor during the entire experiment.

Advances in Application of Biological Nitrification Inhibitors

Based on current research, the characteristics and action mechanism of biological nitrification inhibitors at home and abroad were reviewed by combining with the latest research progress. The application effects of biological nitrification inhibitors on agricultural production were summarized. Research hotspot and achievements of biological nitrification inhibitors at home and abroad were summarized. The research direction in future was forecasted.

Nitrification and Denitrification Activities and N_2O Emission of Orchard Soils Cultivated for Different Periods of Time

[Objective] The aim was to investigate the differences in nitrification and denitrification activities and the N20 emission of orchard soils cultivated for different periods of time. [Method] Incubation experiment was conducted to determine the ni- trification and denitrification activities and N20 emission of three types of orchard soil samples that had been cultivated for 5, 12 and 20 years, respectively, by using the virgin soil sample as control. [Result] After 26 d of incubation, the nitrification rates of nitrogen fertilizer in the virgin soil sample and the orchard soil samples cultivated for 5, 12 and 20 years were 6.85%, 10.26%, 13.29% and 12.90%, respectively, which were positively correlated with content of soil organic matter, ammonium nitro- gen and total nitrogen （P〈0.05）, and negatively correlated with soil carbon-nitrogen ratio and pH value （P〈0.05）. The denitrification activities of these soil samples in- creased with the increase of cultivation years. The amount of nitrogen loss by deni- trification accounted for 0.01%-3.11% of the amount of fertilizer nitrogen, and had a positive correlation with the content of soil organic matter （P〈0.05）. The N20 emis- sions of orchard soil samples were higher than that of the virgin soil samples （P〈 0.05）. [Conclusion] In South China, the nitrification activity of orchard soil is relatively low, but it has a tendency to increase as the cultivation years increases; the denitri- fication activity is relatively high, and increases significantly with the increase of culti- vation years.

Research on the Factors Affecting Nitrification Inhibition of Dicyandiamide(DCD) in Latosol

[Objective] This research aimed at studying the effects of application amount of DCD and physics and chemistry characteristics of soil such as temperature,moisture content,organic matter content and pH on the inhibition of nitrification when applying nitrification inhibitor DCD in latosol.[Method] The experiment was carried out under aerobic incubated conditions.[Result] A significant delay period of nitrification was observed when the application amount of DCD was 10 mg/kg soil,and the inhibition can at least last for 56 d.When the cultural temperature had increased from 10 to 30 ℃,the duration of nitrification inhibition was shortened from 90 to 30 d.The nitrification inhibition of DCD was reduced with the increasing soil moisture content,organic matter content and soil pH,while the duration of nitrification inhibition was prolonged with the decreasing soil moisture content,organic matter content and soil pH.[Conclusion] Nitrification inhibition of DCD can be improved by increasing the application amount or reducing soil temperature,moisture content,organic matter content and pH.

Effects of chlorothalonil and carbendazim on nitrification and denitrification in soils

The effects of chlorothalonil and carbendazim on nitrification and denitrification in six soils in upland and rice paddy environments were investigated. Laboratory aerobic （60% water holding capacity） and anaerobic （flooded） conditions were studied at 25℃ and fungicide addition rates of 5.5 mg/kg A. I. （field rate, FR）, 20 times （20FR） and 40 times （40FR） field rate, respectively. The results indicated that chlorothalonil at the field rate had a slight inhibitory effect on one soil only, and that soil did not nitrify much in the first place. But chlorothalonil at higher rates inhibited nitrification significantly in all soils. For soils JXP and JXU with a pH of less than 5.0, chlorothalonil almost completely stopped their nitrification at 20FR and 40FR during the whole 14 d incubation period. For soils HNP and HNU with a pH of greater than 8.0, chlorothalonil also significantly inhibit nitrification at 20FR and 40FR （p 〈 0.05）. However, NH4＋ that was added to the soil was also almost completely nitrified by the end of the incubation period in these two soils. The effects of chlorothalonil at 20FR and 40FR on the nitrification of JSP and JSU soils, with a pH of 5.4 and 7.2, respectively, were intermediate between the other soil types. Chlorothalonil had no effect on denitrification at the field rate and had little effect at the higher rates of application in some soils. Carbendazim had essentially no effect on nitrification and denitrification in soils assessed.

Determination of respiration, gross nitrification and denitrification in soil profile using BaPS system

A facility of BaPS （Barometric Process Separation） was used to determine soil respiration, gross nitrification and denitrification in a winter wheat field with depths of 0-7, 7--14 and 14-21 cm. N2O production was determined by a gas chromatograph. Crop root mass and relevant soil parameters were measured. Results showed that soil respiration and gross nitrification decreased with the increase of soil depth, while denitrification did not change significantly. In comparison with no-plowing plot, soil respiration increased significantly in plowing plot, especially in the surface soil of 0-7 cm, while gross nitrification and denitrification rates were not affected by plowing. Cropping practice in previous season was found to affect soil gross nitrification in the following wheat-growing season. Higher gross nitrification rate occurred in the filed plot with preceding crop of rice compared with that of maize for all the three depths of 0-7, 7-14 and 14-21 cm. A further investigation indicated that the nitrification for all the cases accounted for about 76% of the total nitrogen transformation processes of nitrification and denitrification and the N2O production correlated with nitrification significantly, suggesting that nitrification is a key process of soil N2O production in the wheat field. In addition, the variations of soil respiration and gross nitrification were exponentially dependent on root mass （p〈0.00l）.

Temperature effect on aerobic denitrification and nitrification

Nitrogen loss without organic removal in biofilter was observed and its possible reason was explained. A lower hydraulic loading could improve aerobic denitrification rate. Aerobic denitrification was seriously affected by low temperature(below 10℃). However, nitrification rate remained high when the temperature dropped from 15℃ to 5℃. It seemed the autotrophic biofilm in BAF could alleviate the adverse effect of low temperature.

Removal of organic matter and nitrogen from distillery wastewater by a combination of methane fermentation and denitrification/nitrification processes

The distillery wastewater of Guangdong Jiujiang Distillery, which is characteristic of containing high organic matters and rich total nitrogen, was treated by a combination of methane fermentation and denitrification/nitrification processes. 80% of COD in the raw wastewater was removed by methane fermentation at the COD volumetric loading rate of 20 kg COD/（m^3·d） using the expanded granule sludge bed （EGSB） process. However, almost all the organic nitrogen in the raw wastewater was converted into ammonia by ammonification there. Ammonia and volatile fatty acids （VFA） remaining in the anaerobically treated wastewater were simultaneously removed utilizing VFA as an electron donor by denitrification occurring in the other EGSB reactor and nitrification using PEG-immobilized nitrifying bacteria with recirculation process. An aerobic biological contact oxidization reactor was designed between denitrification/nitrification reactor for further COD removal. With the above treatment system, 18000-28000 mg/L of COD in raw wastewater was reduced to less than 100 mg/L. Also, ammonia in the effluent of the system was not detected and the system had a high removal rate for 900-1200 mg/L of TN in the raw wastewater, only leaving 400 mg/L of nitrate nitrogen.

Nitrogen removal influence factors in A/O process and decision trees for nitrification/denitrification system

In order to improve nitrogen removal in anoxic/oxic(A/O) process effectively for treating domestic wastewaters, the influence factors, DO(dissolved oxygen), nitrate recirculation, sludge recycle, SRT(solids residence time), influent COD/TN and HRT(hydraulic retention time) were studied. Results indicated that it was possible to increase nitrogen removal by using corresponding control strategies, such as, adjusting the DO set point according to effluent ammonia concentration; manipulating nitrate recirculation flow according to nitrate concentration at the end of anoxic zone. Based on the experiments results, a knowledge-based approach for supervision of the nitrogen removal problems was considered, and decision trees for diagnosing nitrification and denitrification problems were built and successfully applied to A/O process.

Cultivation of aerobic granules for simultaneous nitrification and denitrification by seeding different inoculated sludge

Cultivation of aerobic granules for simultaneous nitrification and denitrification in two sequencing batch airlift bioreactors was studied. Conventional activated floc and anaerobic granules served as main two inoculated sludge in the systems. Morphological variations of sludge in the reactors were observed. It was found that the cultivation of aerobic granules was closely associated with the kind of inoculated sludge. Round and regular aerobic granules were prevailed in both reactors, and the physical characteristics of the aerobic granules in terms of settling ability, specific gravity, and ratio of water containing were distinct when the inoculate sludge was different. Aerobic granules formed by seeding activated floc are more excellent in simultaneous nitrification and denitrification than that by aerobic granules formed from anaerobic granules. It was concluded that inoculated sludge plays a crucial role in the cultivation of aerobic granules for simultaneous nitrification and denitrification.

Effects of Heavy Metals on Ammonification, Nitrification and Denitrification in Maize Rhizosphere

The ammonification, nitrification and denitrification in maize rhizosphere of alluvial soils were compared with those in the bulk soil after exposure to different kinds of heavy metals. The addition of cadmium at low levels (2 mg kg-1 and 5 mg kg-1) could stimulate the ammonification and nitrification in the soils, while inhibition influences were found at high levels of Cd addition (10 mg kg-1 and 20 mg kg-1). The relationship between microbial activity and cadmium concentration varied with the kind of microorganisms. The nitrifying bacteria were more sensitive to cadmium pollution than the ammonifying bacteria. When Cd(II), Cu(II) and Cr(VI) were compared at the same addition concentration of 20 mg kg-1 soil, Cd(II) was the most effective inhibitor of ammonification and denitrification among the three investigated heavy metals, and Cr(VI) had the most strong inhibitory influence on the nitrifying bacteria. The microbial activities in rhizosphere were higher than those in the bulk soil for most of the treatments. Cr(VI) proved to be the most effective in enhancing the microbial activities in rhizosphere, and this could be caused by the positive reduction of Cr(VI) to Cr(III) in rhizosphere, and the relatively sufficient existence of organic matter which intensified the adsorption of the metal. It seemed that the rhizosphere had some mitigation effect on heavy metal toxicity.

Nitrous oxide concentration and nitrification and denitrification in Zhujiang River Estuary,China

The concentrations of nitrous oxide varies between 57 and 329 nmol/dm3, saturation is 674%～4 134% in the Zhujiang River Estuary. This suggests that the area is an emissive source of nitrous oxide. The acetylene inhibition technique is employed to evaluate the rates of nitrification, denitrification and nitrate reduction by bacterial activities in the sediments at three sites. The average of nitrification, denitrification and nitrate reduction rates ranges from 0.32 to 2.43, 0.03 to 0.84 and 4.17 to 13.06 mmol/(m2·h), respectively. The ver- tical profiles of the sediments show that the nitrification and denitrification processes mainly take place in the depth from 0 to 4 cm and depend on regional conditions. The rates of nitrification, denitrification and nitrate reduction are dominated by Eh, nitrate and ammoni- um concentrations in sediments and DO in overlay water. There is a coupling between nitrification and denitrification.

Wastewater Treatment in a Hybrid Biological Reactor (HBR): Nitrification Characteristics

Objective To investigate the nitrifying characteristics of both suspended- and attached- biomass in a hybrid bioreactor. Methods The hybrid biological reactor was developed by introducing porous ceramic particles into the reactor to provide the surface for biomass attachment. Microorganisms immobilized on the ceramics were observed using scanning electron microscopy (SEM). All chemical analyses were performed in accordance with standard methods. Results The suspended- and attached-biomass had approximately the same nitrification activity. The nitrifying kinetic was independent of the initial biomass concentration, and the attached-biomass had a stronger ability to resist the nitrification inhibitor. Conclusion The attached biomass is superior to suspended-biomass for nitrifying wastewater, especially that containing toxic organic compounds. The hybrid biological reactor consisting of suspended- and attached-biomass is advantageous in such cases.

Mitrification－Denitrification Lossof Added Nitrogenin Flooded Rice Rhizosphere

Nitrification-denitrification losses of ￣15N-labelled nitrate and ammonium applied to the rhizosphere andnonrhizosphere of flooded rice were evaluated in 2 greenhouse rhizobox experiments. The loss of added Nvia denitrification was estimated directly by measuring the total fluxes of (N_2O+N_2)￣15N. It was found that 67% and51%-56% of ￣15N-nitrate added to rice rhizosphere were lost as (N_2O+N_)-￣15N in the 2 experiments, respectively,which were comparable to that added to nonrhizosphere soil (70%and47%, respectively), implying that tbedenitrifying activity in rice rhizosphere was as high as that in nonrhizosphere soil. However, only trace amounts(0-0.3% of added N) were recovered as (N_2O+N_2)-￣15N when ￣15N-ammonium was applied to either rhizosphere ornonrhizosphere, which seems to indicate that the nitrifying activity in the either rhizosphere or nonrhizosphere soilswas quite low. The apparent denitrification calculated from ￣15N balance studies was 10%-47% higher than the totalflux of (N_2O+N_2)-￣15N. Reasons for the large differences can not be explained satisfactorily. Though the denitrifyingactivity in rhizospbere was high and comparable to that in nonrhizosphere soil, presumably due to the low nitrifyingactivity and/ or the strong competition of N uptake against denitrification, the nitrification-denitrification takingplace in rhizosphere could not be an important mechanism of loss of ammonium N in flooded rice-soil system.

Simultaneous nitrification and denitrification conducted by Halomonas venusta MA-ZP17-13 under low temperature conditions

Nitrification is a key step in the global nitrogen cycle.Compared with autotrophic nitrification,heterotrophic nitrification remains poorly understood.In this study,Halomonas venusta MA-ZP17-13,isolated from seawater in shrimp aquaculture (Penaeus vannamei),could simultaneously undertake nitrification and denitrification.With the initial ammonium concentration at 100 mg/L,the maximum ammonium-nitrogen removal rate reached98.7%under the optimal conditions including C/N concentration ratio at 5.95,p H at 8.93,and Na Cl at 2.33%.The corresponding average removal rate was 1.37 mg/(L·h)(according to nitrogen) in 3 d at 11.2℃.By whole genome sequencing and analysis,nitrification-and denitrification-related genes were identified,including ammonia monooxygenase,nitrate reductase,nitrite reductase,nitric oxide dioxygenase and nitric oxide synthase;while no gene encoding hydroxylamine oxidase was identified,it implied the existence of a novel nitrification pathway from hydroxylamine to nitrate.These results indicate heterotrophic bacterium H.venusta MA-ZP17-13 can undertake simultaneous nitrification and denitrification at low temperature and has potential for NH_(4)^(+)-N/NH_(3)-N removal in marine aquaculture systems.

Nitrification and denitrification as sources of gaseous nitrogen emission from different forest soils in Changbai Mountain

The contributions of nitrification and denitrification to N2O and N2 emissions from four forest soils on northern slop of Changbai Mountain were measured with acetylene inhibition methods. In incubation experiments, 0.06% and 3% C2H2 were used to inhibit nitrification and denitrification in these soils, respectively. Both nitrification and denitification existed in these soils except tundra soil, where only denitrification was found. The annually averaged rates of nitrification and denitrification in mountain dark brown forest soil were much higher than that in other three soils. In mountain brown coniferous soil, contributions of different processes to gaseous nitrogen emissions were Denitrification N2O>nitrification N2O>Denitrification N2. The same sequence exists in mountain soddy soil as that in the mountain brown coniferous soil. The sequence in mountain tundra soil was Denitrification N2O>Denitrification N2.

Nitrogen Release Kinetics and Nitrification-Denitrification on Surface Sediments under Aerating Disturbance Condition

[Objective] This study aimed to investigate the nitrogen release kinetics and nitrification-denitrification on surface sediments under aerating disturbance condition, with the purpose to solve the sediment nitrogen release and secondary pollution problems. [Method] The effect of in situ sediments aeration on the release of nitrogen pollutants was investigated, and the nitrogen release kinetics parameters were analyzed. The process of nitrification and denitrification under sediments aeration condition was investigated in laboratory. [Result] The nitrogen released from sediments was enhanced by aeration disturbance. The concentration of NH4＋-N and TN reached the maximum value in 30 min, and release rates were proportional to the disturbance strength. In this study, with the distance of aerator to the sediments surface of 0, 1, 2 and 3 cm, the suspended sediments concentrations were 3.52, 3.41, 3.26 and 3.01 g/L, respectively. Maximum release concentration of NH4＋-N and TN were 14.3, 13.8, 13.2, 12.2 mg/L and 33.21, 30.98, 29.83, 27.30 mg/L, respec- tively. In addition, both NH4＋-N and TN release kinetics could be described by Double Constant Equation as InC=A＋Blnt. Nitrification reaction occurred and was promoted by continued aerating to sediments.The concentration of NH4＋-N dropped down from 12.4 mg/L to 0.2 mg/L in 8 d, with the concentration of NO3--N increased to the maximum value of 10.8 mg/L. In addition, concentration of NO3--N and TN decreased from 10.8 mg/L and 37.4 mg/L to 0.36 mg/L and 23.2 mg/L after the stop of aeration for 12 d, indicating the occurrence of denitdfication reaction. Therefore, sediment aeration could accelerate nitrogen release and nitrification reaction, and with intermittent aeration, nitrogen could be removed from sediments in-situ by nitrification and denitrification. [Conclusion] The results provided technical reference for the in situ sediment remediation for the black-odor rivers in cities.