Abundance Of nirS Research Articles

Impacts of Ag and Ag2S nanoparticles on the nitrogen removal within vertical flow constructed wetlands treating secondary effluent.

In this study, the effects of silver (Ag NPs) and sliver sulfide nanoparticles (Ag2S NPs) on nitrogen removal and nitrogen functional microbes in constructed wetlands were investigated. The obtained results demonstrated that inhibition extent on nitrogen removal relied on NPs types and high concentrations NPs showed higher negative effects. 0.5mg/L Ag NPs had no influence on NH4+-N removal, amoA and nxrA gene copies, whereas Ag2S NPs and Ag+ decreased NH4+-N removal by reducing abundances of nitrifying genes. The concentrations of NO3--N and TN in all 0.5mg/L obviously increased compared with control, resulting from decreasing functional genes and denitrifying bacteria. And 0.5mg/L Ag NPs exhibited largest inhibitory effects, with the highest NO3--N effluent concentrations. 2mg/L Ag NPs decreased NH4+-N removal, but adverse effects gradually vanished with extension of time, whereas both Ag2S NPs and Ag+ at 2mg/L influenced NH4+-N transformation and decreased the abundance of amoA and nxrA genes and the AOB Nitrosomonas in CWs. Moreover, 2mg/L of Ag NPs reduced NO3--N removal by decreasing abundance of nirS and key denitrifying bacteria. To sum up, the inhibition mechanisms concluded from current results were possibly in that Ag NPs exhibited nanotoxicity rather than ionic toxicity.

Cd bioavailability and nitrogen cycling microbes interaction affected by mixed amendments under paddy-pak choi continued planting.

Cadmium (Cd) is the most concerning soil pollutant, and a threat to human health, especially in China. The in-situ immobilization of Cadmium by amendments is one of the most widely adopted methods to remedy soil contamination. The study was designed to evaluate the effect of organo-chemical amendments on soil Cd bioavailability and nitrogen cycling microbes under continuous planting of rice (Oryza sativa) and pak choi (Brassica chinensis L.). The experiment was carried out using four amendments, Lime, Zeolite, Superphosphate, and Biochar, at two different ratios; M1: at the ratio of 47:47:5:1, and M2 at the ratio of 71:23:5:1, respectively. Moreover, both M1 and M2 were enriched at four levels (T1: 0.5%; T2: 1%; T3: 2%; T4: 4%). Results showed that compared with CK (Cd enriched soils), the yield of rice under treatments of M1T1 and M2T1 increased by 8.93% and 8.36%, respectively. While the biomass (fresh weight) of pak choi under M1 and M2 amendments increased by 2.52-2.98 times and 0.76-2.89 times respectively, under enrichment treatments T1, T2, and T3. The total Cd concentrations in rice grains treated with M1T3 and M2T3 decreased by 89.25% and 93.16%, respectively, compared with CK. On the other hand, the total Cd concentrations in pak choi under M1T3 and M2T2 decreased by 92.86% and 90.23%, respectively. The results showed that soil pH was the main factor affecting Cd bioavailability in rice and pak choi. The Variance partitioning analysis (VPA) of rice and pak choi showed that soil pH was the most significant contributing factor. In the rice season, the contribution of soil pH (P) on Cd bioavailability was 10.14% (P=0.102), and in the pak choi season, the contribution of soil pH was 8.38% (P=0.133). Furthermore, the abundance of ammonia oxidation and denitrifying microorganisms had significantly correlation with soil pH and exchange Cd. In rice season, when the enrichment level of amendments increased from 0.5% (T1) to 2% (T3), the gene abundance of AOA, AOB, nirK, nirS and nosZ (І) tended to decrease. While in pak choi season, when the enrichment level increased at the level of 0.5% (T1), 1% (T2), and 2% (T3), the gene abundance of AOB, nirS, and nosZ (І) increased. Additionally, the gene abundance of AOA and nirK showed a reduction in the pak choi season contrasting to rice. And the mixed amendment M2 performed better at reducing Cd uptake than M1, which may have correlation with the ratio of lime and zeolite in them. Finally, we conclude that between these two amendments, when applied at a moderate level M2 type performed better than M1 in reducing Cd uptake, and also showed positive effects on both gene abundance and increase soil pH.

Intact and washed biochar caused different patterns of nitrogen transformation and distribution in a flooded paddy soil

Biochar soil amendment has been proposed as a novel way of mitigating greenhouse gas emissions and improving crop productivity. However, the effects of biochar on N2O emissions and rice growth are inconsistent. Here, we investigated how biochar and its residue after washing affected nitrogen (N) transformation and rice growth and how far these effects remain from biochar using novel three-compartment pots. Intact biochar, washed biochar, washed biochar plus extract were placed in the central compartment of the pot, while rice plants were grown in the near-char zone (<5 cm, from the central compartment) or far-char zone (>5 cm). The addition of biochar and its extract generally increased N concentration in the roots and grains for the plants grown in the near-char zone but decreased the N concentrations for those in the far-char zone. The intact biochar increased the aboveground biomass, especially for the plant grown in the near-char zone, whereas the washed biochar had little influence on plant growth at the maturity and on dissolved organic C, total organic N, NH4+-N and available P. However, washed biochar, as well as intact biochar, still lowered N2O emissions from the near- and far-char zones. The treatments, sampling times and rice growth all played important roles in regulating N-related functional genes, while sampling zones only affected the abundances of amoA and nifH genes. The 16S rRNA, bacterial amoA and narG gene abundances were highest with the intact biochar. The abundance of nirS and nirK genes increased with the washed biochar plus its extract but not with washed biochar alone. The N-functional microorganisms responded to the intact biochar, its soluble component and washed biochar differently and were closely correlated to microbial biomass C, NH4+-N, available P and electrical conductivity. Future research should be devoted to studying the effects of soluble component from biochar and the residual effects of biochar loading on plant growth, soil nutrients and N2O emissions.

Low N2O emissions from wheat in a wheat-rice double cropping system due to manure substitution are associated with changes in the abundance of functional microbes

Fertilization has been shown to affect nitrogen (N) cycling and its related functional microbes in agricultural soils. However, the linkage between soil N2O emissions and N-related functional genes under different fertilization strategies from wheat in a wheat-rice double cropping system is rarely examined. Here, we carried out a two-year field study to examine the response of soil N2O emissions driven by N cycling functional genes [archaeal and bacterial amoA (AOA + AOB), nirS, nirK and nosZ] to different fertilization strategies in a wheat cropland of subtropical China. Three fertilizer treatments were established consisting of chemical phosphorus (P) and potassium (K) fertilizer application (PK), chemical N (urea) and PK fertilizer application (NPK), and chemical NPK fertilizer application with chemical N partially replaced with manure (composted pig manure) (NPKM). Over the two wheat seasons, seasonal total N2O emissions averaged 0.66, 3.60 and 3.11 kg N ha−1 for PK, NPK and NPKM plots, respectively. Relative to the NPK treatment, the NPKM treatment significantly decreased N2O emissions by 14 % without compromising grain yield, with a lowered combined fertilizer-induced emission factor (EF) of 1.02 %. Compared with the PK treatment, N fertilization consistently and significantly increased the abundance of ammonium-oxidation bacteria (AOB), nirS, nirK and nosZ genes. The nosZ gene, which drives N2O reduction during denitrification, showed a greater extent under NPKM with manure N combination. The AOB had a more sensitive response than ammonium-oxidation archaea (AOA) to chemical N fertilization. Seasonal N2O emissions showed significant positive correlations with AOB gene abundance and the ratio of (nirK+nirS)/nosZ, while had a negative correlation with nosZ gene abundance across N fertilized treatments. The N2O-related microbial composition of functional genes was significantly changed by N fertilizer application and also showed contrasting patterns between treatments of chemical N fertilizer with and without manure N substitution. Together, partially replacing chemical N fertilizer with manure reduced N2O emissions from wheat in a wheat-rice double cropping system, mainly through decreasing AOB associated-nitrifying potential, and particularly stimulating N2O reduction.

Effects of alpine meadow degradation on nitrifying and denitrifying microbial communities, and N

Meadow degradation is often accompanied by significant changes in nitrogen (N)-cycling and nitrous oxide (N2O) emission potential, and leads to challenges in meadow management. However, the mechanisms of soil N-cycling and N2O emissions remain poorly understood, especially in alpine ecosystems. In this study, we investigated the soil N-cycling process in four alpine meadows on the Tibetan Plateau along a degradation gradient using real-time quantitative polymerase chain reaction and amplicon sequencing to elucidate the mechanisms. Compared to non-degraded meadows, meadow degradation reduced N2O emissions by 38.5–140.2%. Meadow degradation reduced the abundance of amoA of ammonia-oxidising archaea (AOA) and ammonia-oxidising bacteria (AOB), whereas light and moderate degradation increased the abundance of genes nirS, nirK, and nosZ. Lightly degraded meadow exhibited the highest α-diversity of both nitrifiers and denitrifiers, but this higher diversity was not accompanied by higher N2O emissions, with only 32.3% of the microbial taxa identified as predictors of N2O emissions, suggesting that functional redundancy exists in the N-cycling process in meadow ecosystems. Nitrosospira and Mesorhizobium from the AOB and nirK communities, respectively, were identified as the key taxa that may contribute to N2O emissions. Soil properties, especially N reaction substrates, including ammonium-N, nitrate-N, dissolved organic N, and total N, were the primary drivers for N2O emissions via mediation of the N-cycling community, especially nitrifiers. Our results emphasised the importance of environmental factors in shaping nitrifying, denitrifying, and N2O emissions, providing insights for the restoration of degraded meadow ecosystems.

Main controls on the denitrification rates during cropland revegetation in the southwest China Karst Critical Zone Observatory

The revegetation of karst ecosystems in subtropical southwest China that became severely degraded because of intensive agriculture relies on the persistence and recovery of soil functions such as nutrient cycling. Denitrification is a nitrogen (N) removal process that is carried out by nitrifying and denitrifying microorganisms. We still do not have a comprehensive understanding of denitrification and what drives it during revegetation of cropland in degraded karst ecosystems. We used the space-for-time chronosequence method to establish a restoration gradient (sloping cropland, abandoned sloping cropland, and secondary and primary forest) within the Karst Critical Zone Observatory in southwest China. We quantified the abundances of the nitrifier and denitrifier microbial communities and measured the potential denitrification rates (PDR) and basal denitrification rates (BDR) in incubated soils to investigate the denitrification activity in soils at different stages of restoration. The PDR increased through the cropland revegetation phases, and the BDR was lowest in the abandoned sloping cropland (p < 0.05). Both the PDR and BDR were positively correlated with the denitrifier abundances (i.e., nitrite reductase, nirK, and nirS), but were negatively correlated with nitrifier abundances (i.e., ammonia oxidation, AOA, and AOB). The BDR/PDR ratio was 84 % in the sloping cropland but was only 1% in the primary forest, which suggests that the N cycling was inefficient (‘leaky’) in actively managed farmland soils but was ‘tight’ in forest soils. The AOB and nirS abundances explained most of the PDR in the sloping cropland and abandoned sloping cropland, while the soil phosphorus (P) contents explained most of the variation in BDR and PDR in land that was forested in later phases of revegetation. Progressive increases in the PDR as the revegetation progressed was directly controlled by the soil available P and total P contents. Available P was also associated with increases in the carbon (C) and N contents, which influenced the BDR indirectly through increased abundances of nirK and nirS. We conclude that revegetation of cropland helped to facilitate ‘tight’ N-cycling in poor karst soils, and we suggest that judicious P fertilization on sloping cropland would reduce the denitrification activity in degraded ecosystems.

Nir gene-based co-occurrence patterns reveal assembly mechanisms of soil denitrifiers in response to fire.

Denitrification causes nitrogen losses from terrestrial ecosystems. The magnitude of nitrogen loss depends on the prevalence of denitrifiers, which show ecological differences if they harbour nirS or nirK genes encoding nitrite reductases with the same biological function. Thus, it is relevant to understand the mechanisms of co-existence of denitrifiers, including their response to environmental filters and competition due to niche similarities. We propose a framework to analyse the co-existence of denitrifiers across multiple assemblages by using nir gene-based co-occurrence networks. We applied it in Mediterranean soils before and during 1 year after an experimental fire. Burning did not modify nir community structure, but significantly impacted co-occurrence patterns. Bacteria with the same nir co-occurred in space, and those with different nir excluded each other, reflecting niche requirements: nirS abundance responded to nitrate and salinity, whereas nirK to iron content. Prior to fire, mutual exclusion between bacteria with the same nir suggested competition due to niche similarities. Burning provoked an immediate rise in mineral nitrogen and erased the signals of competition, which emerged again within days as nir abundances peaked. nir co-occurrence patterns can help infer the assembly mechanisms of denitrifying communities, which control nitrogen losses in the face of ecological disturbance.

Comparative analysis of upward and downward vertical flow constructed wetlands on the nitrogen removal and functional microbes treating wastewater containing Ag nanoparticles

This study investigated impacts of silver nanoparticles (AgNPs) on nitrogen removal within constructed wetlands (CWs) with different flow directions. The obtained results showed that addition of AgNPs at 0.5 and 2 mg/L significantly inhibited NH4+-N removal, resulting from lower abundances of functional genes (amoA and nxrA) within CWs. And higher abundances of amoA and nxrA genes at 0.5 mg/L were observed in downward flow CW, leading to better NH4+-N removal, compared to upward flow CW. Besides, nitrifying genes amoA and nxrA in upward flow CW at 2.0 mg/L exhibited higher than downward flow CW, explaining better NH4+-N removal in upward flow CW. 0.5 mg/L AgNPs significantly declined NO3−-N and TN removal, resulted from decreasing abundances of nirK, nirS and nosZ. In contrast, abundances of nirK, nirS and nosZ genes had slightly lower or higher than before adding AgNPs in upward flow CW, leading to lower NO3−-N and TN effluent concentrations. High throughput sequencing also indicated the changes of functional bacterial community after exposing to AgNPs.

Shift in nitrogen transformation in peatland soil by nitrogen inputs

Inputs of nitrogen (N) to peatlands in the form of fertilizers have rapidly increased due to the intensification of agricultural systems, impacting ecological processes, and the carbon storage function of peatland. However, detailed information on the impacts of long-term N inputs on the individual steps of N transformation processes in peatland soils still needs to be fully understood. We investigated N mineralization and nitrification rates as well as nitrite dependent anaerobic methane oxidation (n-damo), anaerobic ammonium oxidation (anammox), denitrification, and dissimilatory nitrate reduction to ammonium (DNRA) in a peatland affected by N inputs for >50 years, using isotope tracing technique and quantitative PCR. Based on the results, N inputs increased N mineralization and nitrification rates by 77 and 43%, respectively. Notably, the contributions of n-damo and anammox to N2 production were enhanced by 242 and 170%, accounting for 30 and 12%, respectively. The contributions of denitrification and DNRA to N2 production decreased by 27 and 52%, accounting for 48 and 10% of N2 production, respectively. Nitrifier abundance increased significantly, with AOA being the dominant prokaryote (from 696 to 1090 copies g−1), but AOB responded more strongly to N inputs (from 5 to 68 copies g−1). The N inputs also promoted the growth of n-damo and anammox bacteria, whose abundances increased by 3.7% (from 565 to 586 copies g−1) and 85.7% (from 305 to 567 copies g−1), respectively, while denitrifier abundance was significantly reduced, with nirK and nirS abundances decreasing by 58% (from 738 to 308 copies g−1) and 50% (from 218 to 109 copies g−1), respectively. Soil pH was the key environmental factor influencing N transformations. We show that n-damo plays important roles in N cycling in peatland subjected to N inputs, providing a scientific basis for improved peatland management.

Effects of Sea Animal Activities on Tundra Soil Denitrification and nirS- and nirK-Encoding Denitrifier Community in Maritime Antarctica.

In maritime Antarctica, sea animals, such as penguins or seals, provide a large amount of external nitrogen input into tundra soils, which greatly impact nitrogen cycle in tundra ecosystems. Denitrification, which is closely related with the denitrifiers, is a key step in nitrogen cycle. However, effects of sea animal activities on tundra soil denitrification and denitrifier community structures still have received little attention. Here, the abundance, activity, and diversity of nirS‐ and nirK-encoding denitrifiers were investigated in penguin and seal colonies, and animal-lacking tundra in maritime Antarctica. Sea animal activities increased the abundances of nirS and nirK genes, and the abundances of nirS genes were significantly higher than those of nirK genes (p < 0.05) in all tundra soils. Soil denitrification rates were significantly higher (p < 0.05) in animal colonies than in animal-lacking tundra, and they were significantly positively correlated (p < 0.05) with nirS gene abundances instead of nirK gene abundances, indicating that nirS-encoding denitrifiers dominated the denitrification in tundra soils. The diversity of nirS-encoding denitrifiers was higher in animal colonies than in animal-lacking tundra, but the diversity of nirK-encoding denitrifiers was lower. Both the compositions of nirS‐ and nirK-encoding denitrifiers were similar in penguin or seal colony soils. Canonical correspondence analysis indicated that the community structures of nirS‐ and nirK-encoding denitrifiers were closely related to tundra soil biogeochemical processes associated with penguin or seal activities: the supply of nitrate and ammonium from penguin guano or seal excreta, and low C:N ratios. In addition, the animal activity-induced vegetation presence or absence had an important effect on tundra soil denitrifier activities and nirK-encoding denitrifier diversities. This study significantly enhanced our understanding of the compositions and dynamics of denitrifier community in tundra ecosystems of maritime Antarctica.

Rapid detection of cytochrome cd1-containing nitrite reductase encoding gene nirS of denitrifying bacteria with loop-mediated isothermal amplification assay

The cytochrome cd1-containing nitrite reductase, nirS, plays an important role in biological denitrification. Consequently, investigating the presence and abundance of nirS is a commonly used approach to understand the distribution and potential activity of denitrifying bacteria, in addition to denitrifier communities. Herein, a rapid method for detecting nirS gene with loop-mediated isothermal amplification (LAMP) was developed, using Pseudomonas aeruginosa PAO1 (P. aeruginosa PAO1) as model microorganism to optimize the assay. The LAMP assay relied on a set of four primers that were designed to recognize six target sequence sites, resulting in high target specificity. The limit of detection for the LAMP assay under optimized conditions was 1.87 pg/reaction of genomic DNA, which was an order of magnitude lower than that required by conventional PCR assays. Moreover, it was validated that P. aeruginosa PAO1 cells as well as genomic DNA could be directly used as template. Only 1 h was needed from the addition of bacterial cells to the reaction to the verification of amplification success. The nirS gene of P. aeruginosa PAO1 in spiked seawater samples could be detected with both DNA-template based LAMP assay and cell-template based LAMP assay, demonstrating the practicality of in-field use.

Regulation of the product stoichiometry of denitrification in intensively managed soils

AbstractCrop residue amendment in conjunction with synthetic nitrogen (N) fertilization is a common agricultural practice that increases soil fertility and crop yield. However, such a practice may also change soil denitrification process. Here, we conducted an incubation experiment with a robotized continuous flow N2 free incubation system [using helium (He) and oxygen (O2)] and measured fluxes of N2O and N2 at a high resolution in an intensively managed soil to examine the interaction effect of straw amendment and N fertilization on soil denitrification. Four treatments were set consisting of (a) a nonamended treatment (Control); (b) a Straw treatment (2 g straw kg−1 dry soil); (c) a KNO3 treatment (KNO3, 15 mM KNO3); and (d) a Straw + KNO3 treatment (2 g straw kg−1 dry soil and 15 mM KNO3). During the oxic phase (80% He plus 20% O2) of the experiment (initial 2 days), flux rates of N2O were 0.54 and 0.38 kg N/ha day−1 in the Control and KNO3 treatments, respectively. Meanwhile, straw amendment triggered N2O fluxes immediately after the onset of treatments, which was more evident in the Straw + KNO3 treatment. During the anoxic phase (100% He), both N2O and N2 emissions increased in all treatments, with the effect being more pronounced in the Straw and Straw + KNO3 treatments. In line with the observed differences in gas fluxes, the abundances of nirK, nirS, and nosZ genes increased clearly in these treatments. Overall, the mean N2O/(N2O + N2) ratio (0.69 ± 0.03) in the Control treatment was significantly lower compared to the KNO3 treatment (18.8%) and higher than the straw‐amended soils (31.9% and 17.4% compared to the Straw and Straw + KNO3 treatments, respectively). Taken together, our results suggest straw amendment significantly altered N2O and N2 fluxes by decreasing the N2O/(N2O + N2) ratio; however, the effects of straw amendment depended on soil nitrate content.

Effects of submerged vegetation on sediment nitrogen-cycling bacterial communities in Honghu Lake (China)

Sediment nitrogen (N) cycling is an important biological removal process for N permanently and driven by N-cycling microbial community. There is a growing interest in interactions between submerged vegetation (SV) and sediment N-cycling bacterial community, because of the close link between rooted aquatic plants and the sediment microbes. However, the effects of SV on the sediment N-cycling bacterial community are still controversial. Furthermore, the discrimination of direct and indirect effects of SV on the N-cycling bacterial community remains unclear. Here, we investigated the biomass and species richness of SV and determined the corresponding environment factors (water quality and sediment properties) in Honghu Lake (China). We also used functional genes as markers to unveil the bacterial diversity and community composition and abundance in lake sediments. Our results showed that biomass and species richness of SV affected the composition, diversity and abundance of sediment N-cycling bacterial communities through improving lake water quality and sediment properties. With the increasing richness and abundance of SV, the diversity of most N-cycling bacterial assemblages including nitrifying, denitrifying and DNRA bacteria decreased, while the abundance increased. However, the anammox bacterial assemblage in sediments showed inverse trends. Sediment carbon vs. nitrogen (C:N) ratio negatively affected the abundance of amoA and nirS + nirK + nosZ bacterial assemblages. Additionally, due to the presence of SV, positive interactions among N-cycling bacterial assemblages were found, such as amoA and nrfA bacterial assemblages. Overall, our findings confirmed the significant effects of SV on the N-cycling bacterial community structure and abundance. Moreover, the direct effects of SV on the N-cycling bacterial community and the indirect effects through altering the sediment C were clarified in our study. Our results casted a new light on the negative effects of high C:N ratio. From the study, we made a conclusion that the better SV develops, the greater nitrogen removal occurs in lake sediments.

Efficacy of heated tourmaline in reducing biomass clogging within woodchip bioreactors

Woodchip bioreactors can effectively remove waterborne nitrates from subsurface agricultural drainage and prevent the eutrophication of receiving water, but rapid biofilm growth can severely reduce water flux and denitrification efficiency of this practice within a few years. Tourmaline minerals with thermal excitation could generate reactive oxygen species which would inhibit bacterial growth. In this study, laboratory scale woodchip bioreactors were set up to test the anti-clogging and denitrification efficiency of heated woodchips with tourmaline, heated woodchips without tourmaline, and unheated woodchips. The results showed that the heated tourmaline treatment could reduce the clogging and optimize the nitrate removal rate (47.6 g N/m3/day) under all three hydrologic retention times tested (1, 4, and 8 h). Dissolved oxygen and pH values fluctuated with the removal rate and temperature change, while temperature was identified as the key factor impacting the tourmaline treatment. The heated tourmaline treatment had the lowest biofilm growth (lowest DNA concentration), while the 16S rRNA and a higher abundance of nirS-, nirK-, and nosZ-encoding denitrifying bacteria (based on qPCR) confirmed the higher denitrification efficiency of the heated tourmaline treatment.

Modified cornstalk biochar can reduce ammonia emissions from compost by increasing the number of ammonia-oxidizing bacteria and decreasing urease activity

This study examined how the addition of modified cornstalk biochar (CB) affected ammonia (NH3) emissions during composting. Four treatments were established, including a control (CK) with layer manure and sawdust only, and the CK mixtures adding 10% HNO3 CB (NA), 10% H2O2 CB (HP) and 10% HNO3– H2O2 CB (MI). As the results showed, NH3 emissions was reduced by 47.83% (NA), 61.69% (HP) and 45.69% (MI) when the modified CB used as a compost additive (P < 0.05). According to the data analysis, the addition of modified CB significantly increased the number of ammonia-oxidizing bacteria (AOB), inhibited urease activity and decreased the abundance of narG and nirS at rising temperatures and high temperatures (P < 0.05). Redundancy analysis demonstrated a negative correlation between NH3 emissions and AOB and a positive correlation with urease activity, narG and nirS. Thus, the modified CB helped reduce NH3 emissions by regulating nitrification processes.

Synthetic Fertilizer Increases Denitrifier Abundance and Depletes Subsoil Total N in a Long-Term Fertilization Experiment.

Chronic synthetic nitrogen (N) application can result in a significant accumulation of nitrate in the subsoil, which could alter subsoil N cycle and subsequently affect subsoil N levels. To understand how elemental interactions affect the cycle and storage of subsoil N, we examined the soils receiving no fertilizer control (CK), 30-year applications of synthetic fertilizer (CF), and CF plus organic manure (CF + OM). The N cycling microbial groups and activity were investigated through analyzing abundance of bacteria, nitrifiers and denitrifiers, potential nitrification (PNA) and denitrification (DEA) rates in the topsoil (0–20 cm) and subsoil depths (20–80 cm). Compared with the CK, the CF application increased subsoil nitrate but reduced or did not change subsoil microbial biomass N and total N. Corresponding to the increased nitrate, the abundances of denitrifiers increased in the CF subsoils. By contrast, the abundances of nitrifiers increased in the CF topsoil. Significant correlation between the abundances of nitrifiers and soil PNA was found in the topsoil, while significant correlation was also found in the subsoil between the abundances of nirS- and/or nirK-type denitrifiers and DEA. These results suggest that the depleted or less changed subsoil total N by CF application might be partly related to the enriched denitrifiers groups and the related potential activity. The contrasting responses of nitrifiers and denitrifiers in the CF subsoil indicate a decoupling of both processes. Our findings highlight that the leached nitrate by synthetic fertilizer addition not only occurs as an environmental risk causing groundwater contamination but may also alter the subsoil N cycle through the denitrifier groups.

Nitrogen removal and abundances of associated functional genes in rhizosphere and non-rhizosphere of a vertical flow constructed wetland in response to salinity

It investigated the nitrogen removal performance and the abundances of nitrogen functional genes in the rhizosphere and non-rhizosphere when vertical flow constructed wetland (VFCW) was used to treat brackish eutrophic water. The results demonstrated that both total nitrogen (TN) and ammonia nitrogen (NH4-N) removal efficiencies decreased with the salinity increasing from 0.05% to 1.0%. However, the removal efficiency of nitrate nitrogen (NO3-N) remained more than 93%. It was found by quantitative real-time polymerase chain reaction (qPCR) that abundances of amoA, nosZ, napA, nirK and nirS were greater in the rhizosphere than in the non-rhizosphere at the salinities of 0.05%, 0.5% and 1.0%. As the salinity increased, the abundance of amoA reduced. It indicated that the salinity inhibited the growth of ammonia oxidizing bacteria (AOB) and further hindered the nitrification, which resulted in the deterioration of TN removal performance. The abundances of nosZ, napA, nirK and nirS in the rhizosphere and non-rhizosphere were the highest at 1.0% salinity. The denitrifying bacteria might adapt to the salinity stress, which guaranteed high NO3-N removal efficiency.

Denitrification Potential of Paddy and Upland Soils Derived From the Same Parent Material Respond Differently to Long-Term Fertilization

To investigate the effect of land use and fertilization regimes on soil denitrification potential (SDP), two more than 30-year long-term fertilization experiments derived from same parent material (paddy soil and upland soil) were selected. Generally, the SDP in paddy soil was 6.82 times higher than the upland soil, which was due to the higher abundances of narG, nirS, and nirK genes and nirS-denitrifying bacteria (Bradyrhizobium, Cupriavidus, and Herbaspirillum) in paddy soil. Inorganic fertilization regimes in upland soil did not significantly affect the SDP over Control, while the SDP in NPK and 2NPK of paddy soil decreased by 26.48% and 75.65%, respectively. Compared with Control, NPKOM consistently yielded the highest SDP in both two soils, with 2.47 times and 2.86 times higher for paddy soil and upland soil. The SDP of paddy soil showed correlation with narG and nirS genes mainly regulated by Alo, while the difference in the SDP of upland soil largely depended on the change of nirS-denitrifying bacteria at genus level (Herbaspirillum, Sulfuritalea, and Cupriavidus) and species level, which was mainly controlled by soil pH. PLS path modeling further demonstrated that direct effect of functional genes on the SDP was the greatest for paddy soil, while nirS-denitrifying bacterial communities for upland soil. The results presented herein represent a key step toward understanding the mechanisms that govern SDP under land use and long-term fertilization.

Microbial driving mechanism of biochar and bean dregs on NH3 and N2O emissions during composting

In this study, the effect of biochar (BC) and bean dregs (BD) on nitrifiers and denitrifiers as well as the contributions to the NH3 and N2O emissions were investigated. Compared with the BD treatment, the maximum value of NH3 and N2O emission was decreased by 32.92% and 46.61% in the BD + BC treatment, respectively. The production of NH3 and N2O was closely associated with the abundance and structure of nitrogen functional genes. BD + BC increased the abundance of AOB amoA gene to decrease the NH3 emission. The abundance of nirS was more closely associated with N2O. The abundance of nirS in the BD + BC was lowered by 18.93% compared with the BD treatment, thereby decreasing the N2O emission after composting. Besides, the nosZ-type gene was the more functional denitrification bacterial communities to effect the N2O emissions.

Application of biofertilizer containing Bacillus subtilis reduced the nitrogen loss in agricultural soil

Nitrogen losses caused by excess fertilizer application in agriculture are one of the main sources of non-point pollution. Biofertilizer has been considered as an effective alternative to synthetic nitrogen fertilizer, but the effectiveness and mechanism for controlling non-point pollution remains unclear. In this study, the effect of biofertilizer containing Bacillus subtilis on nitrogen loss in agricultural soil was investigated. Compared with the application of urea alone, the strategy of substituting 50% urea with biofertilizer reduced the nitrogen loss from farmland soil by 54%. Moreover, this strategy also increased nitrogen use efficiency by 11.2% and achieved a 5.0% increase in crop yield. Application of biofertilizer decreased the abundance of bacterial amoA gene and increased the abundance of narG, nirS, nirK, and nosZ genes in soil. This implied a decrease in nitrification and an increase in denitrification. Thus, it reduced the accumulation of NO3−-N in soil and greatly reduced nitrogen runoff and leaching loss. In addition, biofertilizer decreased the abundance of nitrogen-fixing gene nifH by up to 2 times. Application of biofertilizer also increased the abundance of Bacteroidetes and Chloroflexi, which play important roles in degradation of soil organic matter. In conclusion, the biofertilizer regulated the microbial nitrogen transformation process in soil and reduced nitrogen loss from agroecosystems.