Denitrification Potential Research Articles

Responses of Alpine Soil Nitrification and Denitrification Rates to Nitrogen Addition Gradient—The Role of Functional Genes

Increases in nitrogen (N) deposition affect the nitrification and denitrification processes that are regulated by microorganisms. With the aim of understanding what happened to the soil nitrification and denitrification rates under the N application gradients, we set a field experiment treated with N at 6 different rates (0, 2, 4, 8, 16, and 32 g N·m−2·yr−1) in 2014. We determined the physico-chemical properties, abundances and community structures of the nitrifiers and denitrifiers, the net nitrification rate (NNR) and the potential denitrification rate (PDR) of soil samples that were collected in 2020. We found that the abundances of ammonia-oxidizing bacteria amoA (AOB amoA), the sum of ammonia-oxidizing archaea amoA (AOA amoA) and AOB amoA increased with the increase in N application rate, but the abundances of nosZ decreased with that. The microorganisms that dominate the ammonia oxidation process could shift from AOA to AOB under high N application rates. Furthermore, the soil microorganisms respond to the N addition preferentially with the abundance changes rather than the community composition changes. Moreover, the NNR increased with the N input, while the decrease in the PDR was due to the decrease in the pH value caused by high N application. The results also showed that the amoA gene abundance explained most (46.3%) of the variation in the NNR. Moreover, the soil moisture and pH explained 44.0% and 27.1% of the variation in the PDR, respectively. The results demonstrated that the NNR and PDR were mainly explained by functional genes abundances and environmental factors, respectively, in alpine meadow soil under sustained N deposition.

Microbial chemolithoautotrophs are abundant in salt marsh sediment following long-term experimental nitrate enrichment.

Long-term anthropogenic nitrate (NO3-) enrichment is a serious threat to many coastal systems. Nitrate reduction coupled with the oxidation of reduced forms of sulfur is conducted by chemolithoautotrophic microbial populations in a process that decreases nitrogen (N) pollution. However, little is known about the diversity and distribution of microbes capable of carbon fixation within salt marsh sediment and how they respond to long-term NO3- loading. We used genome-resolved metagenomics to characterize the distribution, phylogenetic relationships, and adaptations important to microbial communities within NO3--enriched sediment. We found NO3- reducing sulfur oxidizers became dominant members of the microbial community throughout the top 25cm of the sediment following long-term NO3- enrichment. We also found that most of the chemolithoautotrophic genomes recovered contained striking metabolic versatility, including the potential for complete denitrification and evidence of mixotrophy. Phylogenetic reconstruction indicated that similar carbon fixation strategies and metabolic versatility can be found in several phylogenetic groups, but the genomes recovered here represent novel organisms. Our results suggest that the role of chemolithoautotrophy within NO3--enriched salt marsh sediments may be quantitatively more important for retaining carbon and filtering NO3- than previously indicated and further inquiry is needed to explicitly measure their contribution to carbon turnover and removal of N pollution.

Soil nitrification and denitrification in Cunninghamia lanceolata plantations with different stand ages.

The variations in soil nitrification and denitrification processes, together with the abundances of functional microbes were investigated in Cunninghamia lanceolata plantations with different stand ages of 5, 8, 21, 27, and 40 years old. The results showed that the net nitrification rate fluctuated with increasing forest ages, with that of 8-year- and 27-year-old C. lanceolata plantations being significantly lower than other stand ages. The abundance of ammonia-oxidizing archaea (AOA) amoA in the 27-year-old plantation was significantly lower than that of the 40-year-old plantation, while there was no significant difference among the other stand ages. There was no significant difference in the abundance of AOB amoA gene, denitrifying functional genes or soil denitrification potential among different stand ages. The results of stepwise regression analysis showed that the abundance of AOA amoA gene was not significantly affected by soil physical and chemical properties. In addition, the abundance of AOB was positively associated with soil total carbon content and soil pH. The abundance of denitrifying functional genes including narG, nirK and nosZ increased with increasing soil pH. The abundance of nirK and nirS was influenced by soil total carbon. Stand age influenced soil net nitrification rate through the AOA amoA abundance. Moreover, soil denitrification potential was directly affected by stand age, or indirectly affected by stand age through soil microbial biomass carbon, soil pH and denitrifying gene abundance of narG and nirK. Compared with the denitrification process, soil nitrification and associated AOA amoA gene abundance were more sensitive to the development of C. lanceolata plantations. The rotation period sould be appropriately extended to reduce the risk of nitrogen losses resulting from soil nitrification.

Metagenomic analysis reveals the influence of pH and hydraulic loading on thiosulfate-driven denitratation: Insight into efficient performance and microbial mechanism

This study evaluated the performance of thiosulfate-driven denitratation (TDD) systems. The results demonstrated that a cycle time of 4.00 h was adequate for NO3--N removal and NO2--N accumulation (average NO3--N removal efficiencies (NREs): 95%; average NO3--N accumulation efficiencies (NAEs): 79%) under pH range of 6.50–9.50. According to metagenomic research, the enrichment of dominant denitrifiers is essential for the pH responsiveness of the TDD system. Furthermore, interspecific microorganism cooperation and the potential selective advantage of NO3--N reducing bacteria over NO2--N reducing bacteria contributed to a higher NO2--N accumulation efficiency. In addition, the results revealed that the reduction in hydraulic loading time (HRT) improved metabolic activities (i.e., energy metabolism). Overall, this study broadened the application potential of the TDD system and enhanced the understanding of microbial niches and the metabolic potential of autotrophic denitrification from electron transfer and metagenomic insights.

Groundwater flow in saturated riparian buffers and implications for nitrate removal.

A saturated riparian buffer (SRB) is an edge-of-field conservation practice that intercepts tile drainage and reduces nitrate flux to nearby streams by redistributing the flow as shallow groundwater. In this study, a three-dimensional, finite-difference groundwater model representative of SRBs in central Iowa was developed to assess the flow of groundwater and implications for nitrate removal during spring conditions, when flow to the SRB is highest. The model reproduces field observations of water level with Nash-Sutcliffe efficiency of 0.68, which is deemed acceptable for hydrologic models. The modeling shows that groundwater flow is three-dimensional near the distribution pipe and the stream and primarily one-dimensional in the rest of the buffer. The path the water takes in flowing toward the stream depends on where it exits the distribution pipe. When nitrate is not limiting, the potential for nitrate removal depends on the length of the path-and thus travel time-and depth because denitrification potential varies with depth. Travel time Tt can be estimated well with slight modifications to a one-dimensional approximation: Tt = 1.11Lx /vx , where Lx is the buffer width and vx is a one-dimensional approximation of the average linear velocity of groundwater. Refining knowledge of SRB function is an important step toward enhancing design for improving water quality.

Stormwater biofilter response to high nitrogen loading under transient flow conditions: Ammonium and nitrate fates, and nitrous oxide emissions

Nitrogen (N) in urban runoff is often treated with green infrastructure including biofilters. However, N fates across biofilters are insufficiently understood because prior studies emphasize low N loading under laboratory conditions, or use “steady-state” flow regimes over short time scales. Here, we tested field scale biofilter N fates during simulated storms delivering realistic transient flows with high N loading. Biofilter outflow ammonium (NH4+-N) was 60.7 to 92.3% lower than that of the inflow. Yet the characteristic times for nitrification (days to weeks) and denitrification (days) relative to N residence times (7 to 30 h) suggested low N transformation across the biofilters. Still, across 7 successive storms, total outflow nitrate (NO3−-N) greatly exceeded (3100 to 3900%) inflow nitrate, a result only explainable by biofilter soil N nitrification occurring between storms. Archaeal, and bacterial amoA gene copies (2.1 × 105 to 1.2 × 106 gc g soil−1), nitrifier presence by16S rRNA gene sequencing, and outflow δ18O-NO3− values (-3.0 to 17.1 ‰) reinforced that nitrification was occurring. A ratio of δ18O-NO3− to δ15N-NO3− of 1.83 for soil eluates indicated additional processes: N assimilation, and N mineralization. Denitrification potential was suggested by enzyme activities and soil denitrifying gene copies (nirK + nirS: 3.0 × 106 to 1.8 × 107; nosZ: 5.0 × 105 to 2.2 × 106 gc g soil−1). However, nitrous oxide (N2O-N) emissions (13.5 to 84.3 μg N m − 2 h − 1) and N2O export (0.014 g N) were low, and soil nitrification enzyme activities (0.45 to 1.63 mg N kg soil−1day−1) exceeded those for denitrification (0.17 to 0.49 mg N kg soil−1 day−1). Taken together, chemical, bacterial, and isotopic metrics evidenced that storm inflow NH4+sorbs and, along with mineralized soil N, nitrifies during biofilter dry-down; little denitrification and associated N2O emissions ensue, and thus subsequent storms export copious NO3−-N. As such, pulsed pass-through biofilters require redesign to promote plant assimilation and/or denitrification of mineralized and nitrified N, to minimize NO3−-N generation and export.

Relationships between denitrification rates and functional gene abundance in a wetland: The roles of single- and multiple-species plant communities

Wetland soil denitrification removes excess inorganic nitrogen (N) and prevents eutrophication in aquatic ecosystems. Wetland plants have been considered the key factors determining the capacity of wetland soil denitrification to remove N pollutants in aquatic ecosystems. However, the influences of various plant communities on wetland soil denitrification remain unknown. In the present study, we measured variations in soil denitrification under different herbaceous plant communities including single Phragmites karka (PK), single Paspalum thunbergia (PT), single Zizania latifolia (ZL), a mixture of Paspalum thunbergia plus Phragmites karka (PTPK), a mixture of Paspalum thunbergia plus Zizania latifolia (PTZL), and bare soil (CK) in the Estuary of Nantiaoxi River, the largest tributary of Qingshan Lake in Hangzhou, China. The soil denitrification rate was significantly higher in the surface (0–10 cm) than the subsurface (10–20 cm) layer. Wetland plant growth increased the soil denitrification rate by significantly increasing the soil water content, nitrate concentration, and ln(nirS) + ln(nirK). A structural equation model (SEM) showed that wetland plants indirectly regulated soil denitrification by altering the aboveground and belowground plant biomass, nitrate concentration, abundances of denitrifying functional genes, and denitrification potential. There was no significant difference in soil denitrification rates among PT, PK and ZL. The soil denitrification rate was significantly lower in PTZL than PTPK. Two-plant communities did not necessarily enhance the denitrification rate compared to single planting, the former had a greater competitiveness on N uptake and consequently reduced the amount of nitrate available for denitrification. As PTPK had the highest denitrification rate, co-planting P. thunbergia and P. karka could effectively improve N removal efficiency and help mitigate eutrophication in adjacent aquatic ecosystems. The results of this investigation provide useful information guiding the selection of appropriate wetland herbaceous plant species for wetland construction and the removal of N pollutants in aquatic ecosystems.

Soil texture is an easily overlooked factor affecting the temperature sensitivity of N2O emissions

As a potent greenhouse gas, soil nitrous oxide (N2O) is strongly stimulated by rising temperature, triggering a positive feedback effect of global warming. However, its temperature sensitivity varies greatly among soils with different physical and chemical characteristics, while associated mechanisms remain unknown. Here we performed a meta-analysis of the effect of warming on N2O emission and found distinctions in the response of N2O to temperature increase in soils with different textures. Then, we conducted an incubation experiment on 11 arable soils with varying textures sampled across China. The results show that the temperature sensitivity of N2O emissions was lower as soil texture became more clayey and was consistent with the outcome of meta-analysis. Further analysis was conducted by classifying the soils into clay and loam subgroups. As shown in the clay soil subgroup, N2O emission was significantly correlated with both inorganic nitrogen contents and potential denitrification and nitrification activities. Correlation analysis and partial least square (PLS) path model revealed that temperature mediated N2O emission by regulating nosZ gene abundance indirectly. In loam soils, however, the indirect effect of temperature on N2O production was achieved mainly through nirS gene abundance. Additionally, soil DON content strongly correlated with N2O emission in both subgroups and affected N2O emissions by influencing the abundance of denitrifiers under warming conditions. Our findings suggest that (i) soil texture was an important factor affecting temperature sensitivity of N2O emission and (ii) variable efficacy of warming in soil N2O production might originate from the enriching DON and nitrate content and its different indirect effects on nirS- or nosZ-type denitrifiers.

Simultaneous heterotrophic nitrification and aerobic denitrification potential of Paenibacillus sp. strain GLM-08 isolated from lignite mine waste and its role ammonia removal from mine waste water.

Paenibacillus sp. strain GLM-08 was isolated from a lignite mine waste site in the Barmer basin, Rajasthan, India. The strain is efficient in heterotrophic nitrification and aerobic denitrification. This bacterium could remove approximately more than 95% of NH4+, NO3-, and NO2- in 24 h. The average nitrogen (N) removal rate of the strain was found to be 4.775 mg/L/H, 5.66 mg/L/H, and 5.01 mg/L/H for NH4+, NO3-, and NO2-, respectively. Bioaugmentation of mine wastewater with Paenibacillus sp. strain GLM-08 demonstrated N removal of 86.6% under conditions of a high load of NH4+. The presence of potential genetic determinants (nxrB, nirS, and nosZ) having role in heterotrophic nitrification and aerobic denitrification was confirmed by PCR based analysis. The findings show that this bacterium performs simultaneous nitrification and denitrification and has a high nitrogen removal efficiency indicating the potential application of the strain in the treatment of wastewater.

Brachiaria humidicola Cultivation Enhances Soil Nitrous Oxide Emissions from Tropical Grassland by Promoting the Denitrification Potential: A 15N Tracing Study

Biological nitrification inhibition (BNI) in the tropical grass Brachiaria humidicola could reduce net nitrification rates and nitrous oxide (N2O) emissions in soil. To determine the effect on gross nitrogen (N) transformation processes and N2O emissions, an incubation experiment was carried out using 15N tracing of soil samples collected following 2 years of cultivation with high-BNI Brachiaria and native non-BNI grass Eremochloa ophiuroide. Brachiaria enhanced the soil ammonium (NH4+) supply by increasing gross mineralization of recalcitrant organic N and the net release of soil-adsorbed NH4+, while reducing the NH4+ immobilization rate. Compared with Eremochloa, Brachiaria decreased soil gross nitrification by 37.5% and N2O production via autotrophic nitrification by 14.7%. In contrast, Brachiaria cultivation significantly increased soil N2O emissions from 90.42 μg N2O-N kg−1 under Eremochloa cultivation to 144.31 μg N2O-N kg−1 during the 16-day incubation (p < 0.05). This was primarily due to a 59.6% increase in N2O production during denitrification via enhanced soil organic C, notably labile organic C, which exceeded the mitigated N2O production rate during nitrification. The contribution of denitrification to emitted N2O also increased from 9.7% under Eremochloa cultivation to 47.1% in the Brachiaria soil. These findings confirmed that Brachiaria reduces soil gross nitrification and N2O production via autotrophic nitrification while efficiently stimulating denitrification, thereby increasing soil N2O emissions.

Insight into characteristics of sulphur-based autotrophic denitrifying microbiota in the nitrate removal

ABSTRACT Mariculture wastewater is characterized by low organic carbon to nitrogen ratio (C/N) but high nitrate concentration, which makes it difficult to remove nitrate by the completely heterotrophic denitrification. However, high nitrate discharge poses a threat to the natural environment and human health. Thus, we enriched sulphur-based autotrophic denitrifying (SAD) microbiota and optimized the nitrate removal under different environmental factors and electron donor conditions. The results showed that the dominant genera in the enriched microbiota were previously confirmed autotrophic denitrifiers, Sulfurovum, Thioalkalispira-Sulfurivermis, and Sedimenticola, with a high relative abundance of 41.14%, 21.01%, and 6.17%. Among the environmental factors, pH was the key factor affecting SAD microbiota, and pH 7–9 favoured nitrate removal. However, high pH led to nitrite accumulation (e.g. 10 mg/L at pH = 9), which should be strictly avoided. With regard to electron donors, the optimal concentrations of thiosulphate and nitrate were 50 and 5 mg/L, respectively. The best organic carbon is acetate with an optimal concentration of 10 mg/L. Meanwhile, the initial concentration of thiosulphate was proportional to the nitrate removal rate, while higher concentrations of organic carbon stimulated the heterotrophic denitrification potential of microbiota and thus benefited to dentrification. This study showed that the enriched SAD microbiota was able to achieve efficient nitrate removal under suitable environmental conditions and mixed electron donors and thus presented the potential for application in the treatment of mariculture wastewater.

Trade-offs on carbon and nitrogen availability lead to only a minor effect of elevated CO2 on potential denitrification in soil

The global surplus of reactive nitrogen (Nr) and elevated atmospheric CO2 are both major threats to current and future ecosystem integrity. Denitrification is the primary sink for Nr by the conversion to inert N2 and is also the predominant source of N2O emission. Yet, it is unclear how and to what extent soil denitrification would respond to elevated CO2 (eCO2), which is a major part of the uncertainties on the estimation of terrestrial nitrogen (N) cycle budget under the influence of climate changes. Herein, we provided quantitative results of a global meta-analysis with 127 observations that included data on soil potential denitrification rates (PDR) and denitrifying gene abundance, and 305 observations that included data on N2O emission and soil available N content under eCO2. Averaged across all studies, we found that globally eCO2 had an overall minor effect on soil PDR, but showed a large variation with different edaphic and environmental factors. The response was primarily explained by soil organic carbon (SOC) and mean annual precipitation (MAP), i.e., the higher SOC content or MAP, the lower response of soil PDR to eCO2. Further, eCO2 significantly decreased soil NO3- content (-15.3%) and denitrifying bacteria (-31.3%), but increased nirS (14.7%), napA (24.4%) genes and N2O emission (34.1%). We speculate that eCO2 has both positive and negative effects on soil potential denitrification, i.e., a higher soil C input induced by eCO2 supplies more energy for denitrifiers and thus increases denitrification in N-rich ecosystems, while unbalanced C:N input decreases soil NO3- content, causing substrate deficiency in N-limiting ecosystems under eCO2. Whilst the response of soil PDR to eCO2 is governed by trade-offs on soil C and N availability, this may serve as a buffer on soil available N content in terrestrial ecosystems under eCO2. Collectively, based on our findings, we built a comprehensive conceptual model defining the key regulators, which has significant implications for expanding the understanding and predicting the responses of the terrestrial N cycle to eCO2 in future scenario evaluation.

Conversion of coastal wetland to aquaculture ponds decreased N2O emission: Evidence from a multi-year field study

Land reclamation is a major threat to the world's coastal wetlands, and it may influence the biogeochemical cycling of nitrogen in coastal regions. Conversion of coastal marshes into aquaculture ponds is common in the Asian Pacific region, but its impacts on the production and emission of nitrogen greenhouse gases remain poorly understood. In this study, we compared N2O emission from a brackish marsh and converted shrimp aquaculture ponds in the Shanyutan wetland, the Min River Estuary in Southeast China over a three-year period. We also measured sediment and porewater properties, relevant functional gene abundance, sediment N2O production potential and denitrification potential in the two habitats. Results indicated that the pond sediment had lower N-substrate availability, lower ammonia oxidation (AOA and comammox Nitrospira amoA), nitrite reduction (nirK and nirS) and nitrous oxide reduction (nosZ Ⅰ and nosZ Ⅱ) gene abundance and lower N2O production and denitrification potentials than in marsh sediments. Consequently, N2O emission fluxes from the aquaculture ponds (range 5.4–251.8 μg m–2 h–1) were significantly lower than those from the marsh (12.6–570.7 μg m–2 h–1). Overall, our results show that conversion from marsh to shrimp aquaculture ponds in the Shanyutan wetland may have diminished nutrient input from the catchment, impacted the N-cycling microbial community and lowered N2O production capacity of the sediment, leading to lower N2O emissions. Better post-harvesting management of pond water and sediment may further mitigate N2O emissions caused by the aquaculture operation.

Effects of different drainage conditions on nitrogen losses of an agricultural sandy loam soil

Prolonged waterlogging in agricultural fields has severe consequences for the crop development and growth, and could potentially lead to higher N losses. In this study, a 3.93 ha agricultural field in Denmark was separated into two parts of well-drained (WD) and poorly-drained (PD) based on the installation depth of the tile drains. The field was continuously monitored for drainage, soil water dynamics, nitrogen leaching through the drains, and grain dry matter and nitrogen yields in a 4-year period (2017–2020). Furthermore, denitrification potential of the top 1 m of the soil at both parts of the field was measured through the denitrifying enzyme activity assay, and a 1D Daisy model was utilized to capture the differences between water and nitrogen balances at WD and PD. Results indicated that on average over the 4 years, annual harvested nitrogen in the crops at PD was 14% lower compared to WD, with a significant reduction of 33% in 2017–2018, that coincided with the longest period of waterlogging at PD. Moreover, greater losses of nitrogen through leaching from drainage and other pathways were measured at the PD (109 kg N ha−1 ya−1) compared to the WD (95 kg N ha−1 ya−1). Based on the simulations, losses through preferential flow pathways to the drains dominated at PD and most of the denitrification is expected to occur within the topsoil. Future studies could significantly benefit from monitoring the redox dynamics in the top 30 cm of the PD soils, and increasing the depth of tiles drains by redrainage could reduce the N losses of poorly drained agricultural soils.

Long-term nitrogen addition increases denitrification potential and functional gene abundance and changes denitrifying communities in acidic tea plantation soil

The response of soil denitrification to nitrogen (N) addition in the acidic and perennial agriculture systems and its underlying mechanisms remain poorly understood. Therefore, a long-term (12 years) field trial was conducted to explore the effects of different N application rates on the soil denitrification potential (DP), functional genes, and denitrifying microbial communities of a tea plantation. The study found that N application to the soil significantly increased the DP and the absolute abundance of denitrifying genes, such as narG, nirK, norB, and nosZ. The diversity of denitrifying communities (genus level) significantly decreased with increasing N rates. Moreover, the denitrifying communities composition significantly differed among the soils with different rates of N fertilization. Further variance partitioning analysis (VPA) revealed that the soil (39.04%) and pruned litter (32.53%) properties largely contributed to the variation in the denitrifying communities. Dissolved organic carbon (DOC) and soil pH, pruned litter's total crude fiber (TCF) content and total polyphenols to total N ratio (TP/TN), and narG and nirK abundance significantly (VIP >1.0) influenced the DP. Finally, partial least squares path modeling (PLS-PM) revealed that N addition indirectly affected the DP by changing specific soil and pruned litter properties and functional gene abundance. Thus, the findings suggest that tea plantation is a major source of N2O emissions that significantly enhance under N application and provide theoretical support for N fertilizer management in an acidic tea plantation system.

Neonicotinoid pesticide and nitrate mixture removal and persistence in floating treatment wetlands.

Mesocosm and microcosm experiments were conducted to explore the applicability of floating treatment wetlands (FTWs), an ecologically based management technology, to remove neonicotinoid insecticides and nitrate from surface water. The mesocosm experiment evaluated three treatments in triplicate over a 21-d period. Floating treatment wetland mesocosms completely removed nitrate-N over the course of the experiment even when neonicotinoid insecticides were present. At the completion of the experiment, 79.6% of imidacloprid and degradation byproducts and 68.3% of thiamethoxam and degradation byproducts were accounted for in the water column. Approximately 3% of imidacloprid and degradation byproducts and 5.0% of thiamethoxam and degradation byproducts were observed in above-surface biomass, while ∼24% of imidacloprid and degradation byproducts, particularly desnitro imidacloprid, and<0.1% of thiamethoxam and degradation byproducts were found in the below surface biomass. Further, 1 yr after the experiments, imidacloprid, thiamethoxam, and degradation byproducts persisted in biomass but at lower concentrations in both the above- and below-surface biomass. Comparing the microbial communities of mature FTWs grown in the presence and absence of neonicotinoids, water column samples had similar low abundances of nitrifying Archaeal and bacterial amoA genes (below detection to 104 ml-1 ) and denitrifying bacterial nirK, nirS, and nosZ genes (below detection to 105 ml-1 ). Follow-up laboratory incubations found the highest denitrification potential activities in FTW plant roots compared with water column samples, and there was no effect of neonicotinoid addition (100 ng L-1 ) on potential denitrification activity. Based on these findings, (a) FTWs remove neonicotinoids from surface water through biomass incorporation, (b) neonicotinoids persist in biomass long-term(>1 yr after exposure), and (c) neonicotinoids do not adversely affect nitrate-N removal via microbial denitrification.

The active functional microbes contribute differently to soil nitrification and denitrification potential under long-term fertilizer regimes in North-East China.

Nitrogen (N) cycling microorganisms mediate soil nitrogen transformation processes, thereby affecting agricultural production and environment quality. However, it is not fully understood how active N-cycling microbial community in soil respond to long-term fertilization, as well as which microorganisms regulate soil nitrogen cycling in agricultural ecosystem. Here, we collected the soils from different depths and seasons at a 29-year fertilization experimental field (organic/chemical fertilizer), and investigated the transcriptions of N-cycling functional genes and their contribution to potential nitrification and denitrification. We found that long-term fertilization exerted significant impacts on the transcript abundances of nitrifiers (AOA amoA, AOB amoA and hao) and denitrifiers (narG and nosZ), which was also notably influenced by season variation. The transcriptions of AOA amoA, hao, and narG genes were lowest in autumn, and AOB amoA and nosZ transcript abundances were highest in autumn. Compared to no fertilization, soil potential nitrification rate (PNR) was reduced in fertilization treatments, while soil potential denitrification rate (PDR) was significantly enhanced in organic combined chemical fertilizer treatment. Both PNR and PDR were highest in 0–20 cm among the tested soil depths. Path model indicated active nitrifiers and denitrifiers had significant impact on soil PNR and PDR, respectively. The transcriptions of AOA amoA and nxr genes were significantly correlated with soil PNR (Pearson correlation, r > 0.174, p < 0.05). Significant correlation of napA and nosZ transcriptions with soil PDR (Pearson correlation, r > 0.234, p < 0.05) was also revealed. Random forest analysis showed that SOC content and soil pH were the important factors explaining the total variance of active nitrifers and denitrifiers, respectively. Taken together, long-term fertilization regimes reduced soil PNR and enhanced PDR, which could be attributed to the different responses of active N-cycling microorganisms to soil environment variations. This work provides new insight into the nitrogen cycle, particularly microbial indicators in nitrification and denitrification of long-term fertilized agricultural ecosystems.

Nitrogen Sinks or Sources? Denitrification and Nitrogen Removal Potential in Riparian Legacy Sediment Terraces Affected by Milldams

AbstractRiparian zones are key ecotones that buffer aquatic ecosystems through removal of nitrogen (N) via processes such as denitrification. However, how dams alter riparian N cycling and buffering capacity is poorly understood. Here, we hypothesized that elevated groundwater and anoxia due to the backup of stream water above milldams may enhance denitrification. We assessed denitrification rates (using denitrification enzyme assays) and potential controlling factors in riparian sediments at various depths upstream and downstream of two relict U.S. mid‐Atlantic milldams. Denitrification was not significantly different between upstream and downstream, although was greater per river km upstream considering deeper and wider geometries. Further, denitrification typically occurred in hydrologically variable shallow sediments where nitrate‐N and organic matter were most concentrated. At depths below 1 m, both denitrification and nitrate‐N decreased while ammonium‐N concentrations substantially increased, indicating suppression of ammonium consumption or dissimilatory nitrate reduction to ammonium. These results suggest that denitrification occurs where dynamic groundwater levels result in higher rates of nitrification and mineralization, while another N process that produces ammonium‐N competes with denitrification for limited nitrate‐N at deeper, more stagnant/poorly mixed depths. Ultimately, while it is unclear whether relict milldams are sources of N, limited denitrification rates indicate that they are not always effective sinks; thus, milldam removal—especially accompanied by removal of ammonium‐N rich legacy sediments—may improve riparian N buffering.

Heterogeneity of nitrate reduction indicators across a tile drained agricultural catchment in East Jutland, Denmark

Detailed knowledge of the nitrate (NO3−) attenuation capacity of different landscapes is essential to support current aspirations of locally differentiated fertilizer nitrogen (N) regulation for protection of aquatic ecosystems. In this context, we studied the spatial heterogeneity of NO3−, ammonium (NH4+), and total N concentrations in pore water collected in an artificially drained sub-catchment and complemented this with measurements of apparent electrical conductivity (ECa), potential denitrification rates, and basic soil physico-chemical properties. Piezometer nests were installed in transects to represent a range of ECa and slopes in the sub-catchment and were then categorized according to predetermined classifications to delineate the different redox environments. Water samples were collected (February to August 2017) from piezometers that were installed in nests with screen depths at 25–75 cm, 85–115 cm and 125–165 cm. For transects with an average ECa higher than 20 mS m−1, NO3− concentrations were <5 mg N L−1 at the depth slightly above the drainage depth, whereas transects with an average ECa of <20 mS m−1 had NO3− concentrations ranging from 0.2 to 15 mg N L−1. Transects with ECa higher than 20 mS m−1 had the highest average clay content and estimated water contents and, possibly, higher macropore connectivity across the soil profile, which could enhance NO3− transport and the potential for development of anaerobic microsites facilitating microbial denitrification. Potential denitrification rates correlated with the presence of amorphous and poorly crystalline iron (Fe) oxides, yet the results were inconclusive regarding the possible role of Fe(II) mediated NO3− reduction. By categorizing the piezometer transects according to their average ECa values, patterns of N concentrations were found that reflected differences in nitrate attenuation capacity, thereby supporting the further pursuit of ECa as a simple and operational tool for mapping the variability in nitrate reduction patterns in the unsaturated zone on a sub-catchment scale.

Grazing Intensity Has More Effect on the Potential Nitrification Activity Than the Potential Denitrification Activity in An Alpine Meadow

On the Qinghai–Tibet Plateau, nitrogen (N) cycling, such as nitrification and denitrification, in the alpine meadow soils have been considerably affected by grazing, with possible consequences for nitrous oxide (N2O) emissions. However, there is a lack of understanding about how the potential nitrification activity (PNA) and the potential denitrification activity (PDA) might be affected by the grazing intensity. We collected the soil samples in alpine meadow in the east of the Qinghai–Tibet Plateau that was grazed at different intensities from 2015 in peak growing season 2021. We determined the soil physical and chemical properties, the functional gene abundances of nitrifiers and denitrifiers, and the soil PNA and PDA to explore the relationships between a range of abiotic and biotic factors and the PNA and PDA. We found that the PNA and the nitrifiers were significantly affected by the grazing intensity but that the PDA and the denitrifiers were not. The ammonia-oxidizing archaea (AOA) abundance was highest but the ammonia-oxidizing bacteria (AOB)abundance was lower than the control significantly at the highest grazing intensity. The AOA abundance and the soil NH4+-N explained most of the variation in the PNA. The pH was the main predictor of the PDA and controlled the nirS abundance but not the nirK and nosZ abundances. Overall, the PNA was more responsive to the grazing intensity than the PDA. These findings can improve estimations of the nitrification and denitrification process and N2O emissions in alpine meadow.