Abundance Of nosZ Genes Research Articles

Synergistic effects of heterogeneous mature compost and aeration rate on humification and nitrogen fixing during kitchen waste composting.

Sludge mature compost (SMC) is notable for its high production, easy accessibility, and stable supply. This study investigated the impact of the SMC addition and different aeration rates on the humification and nitrogen fixing process during kitchen waste composting. The results demonstrated that addition of SMC prolonged the thermophilic phase, as a comparison, increased aeration shortened this phase. The addition of SMC and increased aeration enhanced humus formation and nitrogen retention. SMC introduced more amide and polysaccharide compounds into the compost, promoting the Maillard humification pathway. Additionally, both SMC and high aeration inhibited denitrification: the SMC reduced the abundance of the nirK gene, while high aeration decreased the abundance of nosZ gene. Network analysis revealed that higher aeration enhanced fungal interactions but diminished bacterial interactions. Conversely, SMC addition bolstered both bacterial and fungal interactions. The final compost product with SMC addition showed a 11.56%-44.19% reduction in antibiotic resistance gene content compared with the control group, and heavy metal contents remained within safe application limits. The combination of high SMC addition and high aeration achieved optimal humification and nitrogen retention, underscoring its potential as a promising solution for kitchen waste composting.

Freshwater snails (Bellamya aeruginosa) bioturbation to enhance nitrogen removal and associated mechanism in constructed wetlands

In this study, total nitrogen (TN) removal efficiency of Pontederia cordata and Myriophyllum elatinoides in surface flow constructed wetlands (SFCWs) with Bellamya aeruginosa were 6.43% and 3.54% higher, respectively, than those in non-B. aeruginosa SFCWs. Further, bioturbation could promote N uptake by plants and release from sediment. In summer and autumn, potential nitrification rate was significantly higher in SFCWs with snails than that in SFCWs without snails. In each season, potential denitrification rate was significantly higher in SFCWs with snails than that in SFCWs without snails. Additionally, ammonia oxidizing archaea, narG, nirS, nirK and nosZ gene abundances were significantly higher in SFCWs with snails than those in SFCWs without snails. Structural equation model analysis revealed a strong positive correlation between nitrifiers and denitrifiers in SFCWs with snails, suggesting that bioturbation enhanced N removal by increasing synergistic effect of nitrifying and denitrifying bacteria.

Combined metabolomic and microbial community analyses reveal that biochar and organic manure alter soil C-N metabolism and greenhouse gas emissions

The use of biochar to reduce the gas emissions from paddy soils is a promising approach. However, the manner in which biochar and soil microbial communities interact to affect CO2, CH4, and N2O emissions is not clearly understood, particularly when compared with other amendments. In this study, high-throughput sequencing, soil metabolomics, and quantitative real-time PCR were utilized to compare the effects of biochar (BC) and organic manure (OM) on soil microbial community structure, metabolomic profiles and functional genes, and ultimately CO2, CH4, and N2O emissions. Results indicated that BC and OM had opposite effects on soil CO2 and N2O emissions, with BC resulting in lower emissions and OM resulting in higher emissions, whereas BC, OM, and their combined amendments increased cumulative CH4 emissions by 19.5 %, 31.6 %, and 49.1 %, respectively. BC amendment increased the abundance of methanogens (Methanobacterium and Methanocella) and denitrifying bacteria (Anaerolinea and Gemmatimonas), resulting in an increase in the abundance of mcrA, amoA, amoB, and nosZ genes and the secretion of a flavonoid (chrysosplenetin), which caused the generation of CH4 and the reduction of N2O to N2, thereby accelerating CH4 emissions while reducing N2O emissions. Simultaneously, OM amendment increased the abundance of the methanogen Caldicoprobacter and denitrifying Acinetobacter, resulting in increased abundance of mcrA, amoA, amoB, nirK, and nirS genes and the catabolism of carbohydrates [maltotriose, D-(+)-melezitose, D-(+)-cellobiose, and maltotetraose], thereby enhancing CH4 and N2O emissions. Moreover, puerarin produced by Bacillus metabolism may contribute to the reduction in CO2 emissions by BC amendment, but increase in CO2 emissions by OM amendment. These findings reveal how BC and OM affect greenhouse gas emissions by modulating soil microbial communities, functional genes, and metabolomic profiles.

Evaluating the role of high N2O affinity complete denitrifiers and non-denitrifying N2O reducing bacteria in reducing N2O emissions in river

Freshwater rivers are hotspots of N2O greenhouse gas emissions. Dissolved organic carbon (DOC) is the dominant electron donor for microbial N2O reduction, which can reduce N2O emission through enriching high N2O affinity denitrifiers or enriching non-denitrifying N2O-reducing bacteria (N2ORB), but the primary regulatory pathway remains unclear. Here, field study indicated that high DOC concentration in rivers enhanced denitrification rate but reduced N2O flux by improving nosZ gene abundance. Then, four N2O-fed membrane aeration biofilm reactors inoculated with river sediments from river channel, estuary, adjacent lake, and a mixture were continuously performed for 360 days, including low, high, and mixed DOC stages. During enrichment stages, the (nirS+nirK)/nosZ ratio showed no significant difference, but the community structure of denitrifiers and N2ORB changed significantly (p < 0.05). In addition, N2ORB strains isolated from different enrichment stages positioned in different branches of the phylogenetic tree. N2ORB strains isolated during high DOC stage showed significant higher maximum N2O-reducing capability (Vmax: 0.6 ± 0.4 ×10−4 pmol h−1 cell−1) and N2O affinity (a0: 7.8 ± 7.7 ×10−12 L cell−1 h−1) than strains isolated during low (Vmax: 0.1 ± 0.1 ×10−4 pmol h−1 cell−1, a0: 0.7 ± 0.4 ×10−12 L cell−1 h−1) and mixed DOC stages (Vmax: 0.1 ± 0.1 ×10−4 pmol h−1 cell−1, a0: 0.9 ± 0.9 ×10−12 L cell−1 h−1) (p < 0.05). Hence, under high DOC concentration conditions, the primary factor in reducing N2O emissions in rivers is the enrichment of complete denitrifiers with high N2O affinity, rather than non-denitrifying N2ORB.

Converse Responses of Biochar Application on N2O Emissions in Soils at Different pH Values in a Subtropical Citrus Orchard

The aim of this study was to explore the effect of biochar on N2O emissions in soils with different pH levels. Soils with five pH levels (4.0, 5.1, 5.8, 6.6, and 7.2) were incubated in two conditions, with 0% biochar (CK) and 1% biochar (BC), for 23 days. N2O emissions were measured at nine time points, and soil chemical properties, AOA-amoA, AOB-amoA, nirK, nirS, and nosZ, were analyzed. Partial least squares path modelling (PLS-PM) was used to assess the effect of nitrification and denitrification pathways on potential N2O emissions. The results showed that biochar reduced N2O emissions in highly acidic soil (pH 4.0) but increased emissions in soils with pH values ranging from 5.1 to 7.2. In highly acidic soils, decreased N2O emission was associated with increased soil pH (p &lt; 0.05) and decreased dissolved organic carbon content (p &lt; 0.05), leading to higher nosZ gene abundance (p &lt; 0.05). Meanwhile, in acidic to neutral soils, biochar application increased soil pH (6.6–11.7%), dissolved organic nitrogen (5.9–29.5%), dissolved organic carbon (8.6–41.0%), stimulated AOB-amoA, nirK, nirS gene abundance (p &lt; 0.05), and thus increased N2O emissions. The results verified the influence of nitrification and denitrification genes on N2O production in soils with different pH values. In conclusion, biochar had different effects on N2O emissions based on soil pH, highlighting the need to consider pH when using biochar to mitigate N2O emissions in subtropical citrus orchards.

The electrochemical mechanism of biochar for mediating the product ratio of N2O/(N2O + N2) in the denitrification process

The biochar electrochemical properties and surface functional groups significantly impact N2O production and reduction during denitrification process. However, its effects on N2O emissions during the denitrification process and its electrochemical mechanisms remain unclear. The study examined the impact of pristine and oxidized biochar combined with two types of nitrogen fertilizers on the N2O/(N2O + N2) ratio and N2O emissions in an incubation experiment with seven treatments: (1) CK (no application of chemical fertilizer); (2) N1 (applying (NH4)2SO4); (3) N1B ((NH4)2SO4 + pristine biochar); (4) N1BO ((NH4)2SO4 + oxidized biochar); (5) N2 (applying KNO3); (6) N2B (KNO3 + pristine biochar); (7) N2BO (KNO3 + oxidized biochar). The study found that in comparison with applying nitrogen fertilizer alone, combining pristine biochar decreased soil N2O concentration by 7.1 %–85.8 %, while combining oxidized biochar increased it by 15.7 %–125.6 %. Applying pristine biochar reduced N2O/(N2O + N2) ratio by 10.4 %–86.2 %, whereas applying oxidized biochar increased it by 12.9 %–121.6 %. The application of pristine biochar increased the nosZ gene abundance and decreased the (nirS + nirK)/nosZ ratio, which contributed to reducing N2O to N2. Compared with oxidized biochar, the oxygen-containing functional groups of pristine biochar decreased by 46.6 %, and it possessed a higher specific surface area (23.01 m2 g−1) and electrical conductivity (0.003 mS cm−1). The correlation analysis showed that DOC and inorganic nitrogen were the key environmental factors affecting N2O emissions. Additionally, the electrical conductivity, specific capacitance, and oxygen-containing functional groups of the biochar were identified as the main factors driving N2O emissions. The SEM analysis suggested that the indirect influence of biochar electrochemical properties on N2O emissions was greater than its direct influence. Our work provides fresh perspectives on reducing soil N2O emissions and establishes a theoretical foundation for the subsequent preparation of biochar materials with enhanced N2O reduction capabilities.

Rice-crayfish integrated system enhances global warming potential via increasing methane emission mainly driven by continuous deep flooding

The conversion from rice monoculture (RM) to rice-crayfish integrated system (RCIS) could change soil physicochemical properties and microorganisms related to greenhouse gas (GHG) emissions. Nevertheless, it is still unclear the responses of GHG emissions and global warming potential (GWP) from paddy fields to RCIS. Here, we conducted a field experiment to investigate the changes in the emissions of methane (CH4) and nitrous oxide (N2O), GWP, soil physicochemical properties and the associated microbial abundances (methanogen, methanotrophs, denitrifier and nitrifier) under RCIS in Gaoyou City, Jiangsu Province, China from 2022 to 2023. There were three treatments, including RM, rice monoculture with continuous deep flooding (RMF) and RCIS. Compared with RM, RCIS and RMF all significantly increased CH4 emission and decreased N2O emission. But the effectiveness of increasing CH4 emission and decreasing N2O emission was larger under RCIS than RMF. The increased CH4 emission was due to the significantly higher mcrA gene abundance and ratio of mcrA/pmoA under RCIS. While the significantly higher nosZ gene abundance and ratio of nosZ/(amoA+nirK+nirS) under RCIS were responsible for its reduced N2O emission. Furthermore, the increases in gene abundances of mcrA and nosZ under RCIS were closely correlated with the significantly lower redox potential and pH as well as the significantly higher contents of dissolved organic carbon, ammonium and nitrate in soil. Averaged across two years, GWP under RCIS and RMF were 4.6-fold and 3.4-fold that under RM, respectively. This revealed that continuous deep flooding had a greater effectiveness in increasing GWP than crayfish culture under RCIS. Notably, the contribution rates of CH4 emission to the total GWP decreased in the order of RCIS (98.6%) > RMF (97.5%) > RM (86.2%). Besides, greenhouse gas intensity under RCIS was 5.1-fold that of RM due to its enhanced GWP and reduced rice grain yield. In summary, RCIS could enhance GWP from paddy fields through increasing CH4 emission mainly caused by the continuous deep flooding.

New insights for simultaneous nitrogen removal and greenhouse gas reduction by biological-electrolysis constructed wetland in the treatment of aquaculture wastewater under varied C/N

Aiming to address the challenges of poor nitrogen (N) removal and greenhouse gas (GHG) emissions in aquaculture wastewater treatment, biological-electrolysis constructed wetland (BECW) and constructed wetland (CW) were established to comparatively evaluate the N removal efficiency, the greenhouse gas (GHG) emissions, and specific microbial gene abundances under various influent C/N ratios. Increasing influent C/N ratio markedly enhanced the removal rates of COD, NH4+-N, NO3−-N, and TN, while markedly reduced N2O fluxes in CW and BECW. The integration of an electric field with CW (BECW) significantly compensated for the shortcomings of the poor NO3−-N removal efficiency and high N2O emissions when C/N < 6. However, this compensating effect gradually weakened with the continue increase of C/N. BECW simultaneously achieved the high COD (89.26 ± 6.08 %), NO3−N (97.03 ± 1.22 %), and TN (83.17 ± 3.19 %) removal rates, and low GHG emissions (486.00 ± 36.61 mg/m2/h) at C/N = 6. The BECW significantly decreased the abundance of mcrA and pmoA genes while increased the abundance of nirK, nirS, and nosZ genes, and the ratios of pmoA/mcrA and nosZ/(nirS + nirK) across all C/N condition. The emission flux of CH4 in BECW was positively correlated with C/N and the mcrA abundance but significantly negatively correlated with pmoA abundance and pmoA/mcrA. The emission flux of N2O was positively correlated with nirS and nirK abundance but significantly negatively correlated with C/N, NO3−-N removal rates, nosZ abundance, and the index of nosZ/(nirS + nirK). This research offers an innovative approach to get both low GHG emissions and high NO3−-N removal rate in aquaculture wastewater treatment.

Deficit irrigation interacting with biochar mitigates N2O emissions from farmland in a wheat–maize rotation system

Biochar application to agricultural fields is an effective carbon sequestration measure that has the potential to reduce N2O emissions and increase soil water holding capacity. However, the interaction mechanisms of biochar under deficit irrigation on N2O emissions remain unclear. A two-year field experiment is conducted in the Guanzhong Plain, China, in order to quantify the effects of biochar and deficit irrigation on N2O emissions from winter wheat–summer maize crop rotation and to investigate the potential mechanisms of nitrification and denitrification. According to the combination of biochar application and actual evapotranspiration-based irrigation scheduling, four treatments are designed (B1W100: biochar 30 t·ha−1 + ET; B1W80: biochar 30 t·ha−1+ 0.8 ET; B0W100: no biochar + ET; B0W80: no biochar + 0.8ET). The soil N2O flux, soil physical and chemical properties, and key functional gene abundance related to N2O emissions in nitrification and denitrification at different growth stages are investigated and discussed. Results show that the interaction between deficit irrigation and biochar significantly reduces soil N2O emissions. During the wheat and maize season, the application of biochar reduces the N2O emissions by an average of 12.9% and 15.2%, respectively. Deficit irrigation also reduces the N2O emissions by an average of 17.4% and 15.5%, respectively. Pearson correlation analysis shows that soil N2O is significantly correlated with soil water-filled pore space during the phase with intense N2O emissions. Soil functional gene abundance is determined at different growth stages for both wheat and maize. Maximum soil denitrification functional gene abundance is observed at the time when wheat and maize enter the stage of their peak growth at the jointing stage. With biochar addition and deficit irrigation, the abundance of nirK and nosZ genes increases and AOB amoA genes decreases. These results suggest that biochar with deficit irrigation is a better solution to reduce N2O emissions from agricultural soils.

Agricultural fertilization near marshes impacts the potential for greenhouse gas emissions from wetland ecosystems by modifying microbial communities

Ensuring agricultural security and preserving the health of wetland ecosystems are crucial concerns facing northeast China. However, the adverse effects of environmental pollution, especially nitrogen (N), caused by prolonged agricultural development on the health of marsh wetlands cannot be systematically recognized. To address this issue, an 18-year trial with four different levels of N application was carried out in a typical area of the Northeast region: 0, 6, 12, and 24 gN·m−2·a−1 (referred to as CK, N6, N12, and N24, respectively) to investigate changes in wetland ecological functioning. The results showed that long-term N input significantly enhanced soil N availability. High-level of N addition (N24) significantly reduced soil bacterial richness in October, while fungal diversity was significantly higher in June than in October for both control and N6 treatments. The main environmental factors affecting microorganisms in June were TN, NH4+, and EC, while bacterial and fungal communities were influenced by TN and Leaf Area Index (LAI), respectively, in October. It was found that the AN16S gene was significantly higher in June than in October, indicating that summer is the critical time for N removal in the wetland. N addition significantly reduced the abundance of the NIFH gene and decreased the N fixation potential of the wetland. In June, low and medium levels of N inputs promoted denitrification processes in the wetland and elevated the wetland N2O emission potential. The abundance of NARG, NIRK, and NOSZ genes decreased significantly in October compared to June, indicating a decrease in the wetland N2O emission potential. Additionally, it was observed that soil methanotrophs were positively affected by NH4+ and TN in October, thereby reducing the wetland CH4 emission potential. Our research provides a systematic understanding of the impact of agricultural N pollution on marsh wetlands, which can inform strategies to protect wetland health.

Regulation of nitrogen transformation and microbial community by inoculation during livestock manure composting.

This study examined the effects of three Bacillus strains and one Saccharomyces cerevisiae strain on nitrogen transformation and microbial communities in pig and chicken manure compost. The findings revealed that the use of compound microbial inoculants increased the compost temperature, accelerated moisture reduction, enhanced cellulase activity, and stimulated the accumulation of NH4 +-N, NO3 --N, and total nitrogen (TN), resulting in a 9% increase in TN content. The abundance of Firmicutes decreased by 3.95% at the maturation phase, while Actinobacteria and Bacteroidetes increased by 1.64% and 1.85%, respectively. Inoculation led to an increase in amoA, nxrA and nifH gene copy numbers, while simultaneously reducing the abundance of nirK, nosZ and nirS genes. It also resulted in an increase in functional enzyme levels, specifically nif and amo, with a corresponding decrease in nor. Clostridium, Phascolarctobacterium, Eubacterium and Faecalibacterium from the class Clostridium, which have a significant correlation with nifH and nxrA genes, suggest their likely crucial role in nitrogen retention and fixation. Inoculation aided in the removal of pathogenic bacteria and antibiotic resistance genes (ARGs) like fluoroquinolones, nucleosides and nitroimidazole. This study provides effective theoretical support for the mechanism of nitrogen retention and fixation, and for improving the quality of compost.

Shallow wet irrigation reduces nitrogen leaching loss rate in paddy fields by microbial regulation and lowers rate of downward migration of leaching water: a 15N-tracer study.

China consumes 35% of the world's fertilizer every year; however, most of the nitrogen fertilizers, which are essential for rice cultivation, are not used effectively. In this study, factors affecting the nitrogen leaching loss rate were studied in typical soil and rice varieties in South China. The effects of various irrigation measures on rice growth and nitrogen leaching loss were investigated by conducting experiments with eight groups. These groups included traditional irrigation (TI) and shallow wet irrigation (SWI). The TI is a common irrigation method for farmers in South China, maintaining a water layer of 5-8cm depth. For SWI, after establishing a shallow water layer usually maintaining at 1-2cm, paddy is irrigated when the field water level falls to a certain depth, then this process is then repeat as necessary. The nitrogen distribution characteristics were determined using 15N isotope tracing. In addition, the effects of nitrification, denitrification, and microbial composition on soil nitrogen transformation at different depths were studied by microbial functional gene quantification and high-throughput sequencing. The results revealed that in the SWI groups, the total nitrogen leaching loss rate reduced by 0.3-0.8% and the nitrogen use efficiency (NUE) increased by 2.18-4.43% compared with those in the TI groups. After the 15N-labeled nitrogen fertilizer was applied, the main pathways of nitrogen were found to be related to plant absorption and nitrogen residues. Furthermore, paddy soil ammonia-oxidizing archaea were more effective than ammonia-oxidizing bacteria for soil ammonia oxidation by SWI groups. The SWI measures increased the relative abundance of Firmicutes in paddy soil, enhancing the ability of rice to fix nitrogen to produce ammonium nitrogen, thus reducing the dependence of rice on chemical fertilizers. Moreover, SWI enhanced the relative abundance of nirS and nosZ genes within surface soil bacteria, thereby promoting denitrification in the surface soil of paddy fields. SWI also promoted ammonia oxidation and denitrification by increasing the abundance and activity of Proteobacteria, Nitrospirae, and Bacteroidetes. Collectively, SWI effectively reduced the nitrogen leaching loss rate and increase NUE.

The Impact of Bacillus coagulans X3 on Available Nitrogen Content, Bacterial Community Composition, and Nitrogen Functional Gene Levels When Composting Cattle Manure

Nitrogen loss is an unavoidable problem during organic waste composting, while exogenous microbial inoculation is a promising strategy for reducing nitrogen loss and improving compost quality. This study was designed to probe available nitrogen levels, bacterial community composition, and the levels of nitrogen functional genes present when composting cattle manure with or without the addition of Bacillus coagulans X3. Bacterial supplementation was associated with the prolongation of the thermophilic stage and improved maturity of the resultant compost. At the maturity stage, samples to which B. coagulans X3 had been added exhibited significant increases in ammonium nitrogen, nitrate nitrogen, and total nitrogen levels. The dominant bacterial phyla observed in these composting samples were Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteriota, and Chloroflexi. B. coagulans X3 addition resulted in significant increases in relative Firmicutes abundance during the thermophilic and cooling stages while also increasing amoA and nosZ gene abundance and reducing nirS gene levels over the course of composting. Together, these data suggest that B. coagulans X3 supplementation provides an effective means of enhancing nitrogen content in the context of cattle manure composting through the regulation of nitrification and denitrification activity.

The effects and mechanisms of deep straw incorporation and denitrifying bacterial agents on mitigating nitrate leaching and N2O emissions in four soil types in the North China Plain

The agricultural practice of straw incorporation is often employed to enhance soil fertility, improve soil properties and reduce nitrate leaching via deep burying. Nevertheless, the tradeoff between nitrate leaching and N2O emissions caused by deep straw incorporation in various soil types remains uncertain. To address this knowledge gap, a mesocosm column experiment was established to evaluate the impact of straw amendment (NS) and of straw plus application of denitrifying bacterial agents (NSS) at column depths of 20–60 cm on NO3--N leaching and N2O emission across four soil types (brown, cinnamon, fluvo-aquic and mortar black soil) in the North China plain. The results showed that the NS and NSS treatments significantly reduced NO3--N leaching by 47%–73% and 37%–81%, respectively, across four soil types, in comparison to the control without straw and bacterial agents (N), with the greatest reduction recorded in the mortar black soil (by 73% for NS and 81% for NSS). Meanwhile, the NS and NSS treatments resulted in a significant increase in cumulative N2O emissions (by 289% for NS and 598% for NSS) in the acidic brown soil with a soil pH ranging between 4.8 and 6.5, but showed no significant effect in the remaining three soil types in which soil pH is above 7.5. When compared to the NS treatment, NSS treatment reduced the cumulative N2O emissions in the fluvo-aquic soil by 42%. We further showed that soil type is the primary driver of bacterial community composition (R2 = 62.3%; P < 0.001), followed by soil depth (R2 = 5.2%; P < 0.001). The incorporation of straw and denitrifying bacterial agents induced significant change in the bacterial community in the middle depth layers, and increases in nosZ gene abundance and the relative abundance of inoculant-similar OTUs significantly by 0.5–37 folds and 2.8–6900 folds, respectively, in all soil types, suggesting the colonization of denitrifying bacterial agents. Our results further suggested that the Streptomycetales were the most important predictors for N2O emissions, and the relative abundance of Streptomycetales was highly positively correlated with N2O emissions across the four soil types. Overall, the study suggests that deep straw incorporation as soil amendment is effective in mitigating nitrate leaching in all four soil types, especially in the fluvo-aquic soil, due to its dual effects on controlling nitrate leaching and N2O emissions. However, it is not recommended for the acidic brown soils as it may cause increased N2O emissions.

Mitigation of N2O emissions in water-saving paddy fields: Evaluating organic fertilizer substitution and microbial mechanisms

Water-saving irrigation strategies can successfully alleviate methane emissions from rice fields, but significantly stimulate nitrous oxide (N2O) emissions because of variations in soil oxygen level and redox potential. However, the relationship linking soil N2O emissions to nitrogen functional genes during various fertilization treatments in water-saving paddy fields has rarely been investigated. Furthermore, the mitigation potential of organic fertilizer substitution on N2O emissions and the microbial mechanism in rice fields must be further elucidated. Our study examined how soil N2O emissions were affected by related functional microorganisms (ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), nirS, nirK and nosZ) to various fertilization treatments in a rice field in southeast China over two years. In this study, three fertilization regimes were applied to rice cultivation: a no nitrogen (N) (Control), an inorganic N (Ni), and an inorganic N with partial N substitution with organic manure (Ni+No). Over two rice-growing seasons, cumulative N2O emissions averaged 0.47, 4.62 and 4.08 kg ha−1 for the Control, Ni and Ni+No treatments, respectively. In comparison to the Ni treatment, the Ni+No fertilization regime considerably reduced soil N2O emissions by 11.6% while maintaining rice yield, with a lower N2O emission factor (EF) from fertilizer N of 0.95%. Nitrogen fertilization considerably raised the AOB, nirS, nirK and nosZ gene abundances, in comparison to the Control treatment. Moreover, the substitution of organic manure for inorganic N fertilizer significantly decreased AOB and nirS gene abundances and increased nosZ gene abundance. The AOB responded to N fertilization more sensitively than the AOA. Total N2O emissions significantly correlated positively with AOB and nirS gene abundances while having a negative correlation with nosZ gene abundance and the nosZ/nirS ratio across N-fertilized plots. In summary, we conclude that organic manure substitution for inorganic N fertilizer decreased soil N2O emissions primarily by changing the soil NO3−-N, pH and DOC levels, thus inhibiting the activities of ammonia oxidation in nitrification and nitrite reduction in denitrification, and strengthening N2O reduction in denitrification from water-saving rice paddies.

Comparative analysis of three next-generation sequencing techniques to measure nosZ gene abundance in Missouri claypan soils

Quantitative next-generation sequencing techniques have been critical in gaining a better understanding of microbial ecosystems. In soils, denitrifying microorganisms are responsible for dinitrogen (N2) production. The nosZ gene codes for nitrous oxide reductase, the enzyme facilitating the reduction of nitrous oxide (N2O) to N2. The objectives of this research were to: 1) understand how soil depth influences RNA concentration and nosZ gene abundance; 2) assess the spatial dependence of nosZ gene abundance in two claypan soil fields; and 3) compare and evaluate multiple RNA-based sequencing methods for quantifying nosZ gene abundance in soils in relation to dinitrogen (N2) production. Research sites consisted of two intensively studied claypan soil fields in Central Missouri, USA. Soil cores were collected from two landscape transects across both fields and analyzed for extractable soil RNA at two depths (0–15 cm and 15–30 cm). Measurements of nosZ gene abundance were obtained using real-time quantitative polymerase chain reaction (RT-qPCR), droplet digital polymerase chain reaction (ddPCR), and nanostring sequencing (NS). In both fields, soil RNA concentrations were significantly greater at 0–15 cm depth compared to 15–30 cm. These data indicated low overall soil microbial activity below 15 cm. Due to low quantities of extractable soil RNA in the subsoil, nosZ gene abundance was only determined in the 0–15 cm depth. Sequencing method comparisons of average nosZ gene abundance showed that NS results were constrained to a narrow range and were 10-20-fold lower than ddPCR and RT-qPCR at each landscape position within each field. Droplet digital PCR appears to be the most promising method, as it reflected changes in N2 production across landscape position.

Introduction of soybean into maize field reduces N2O emission intensity via optimizing nitrogen source utilization

The application of synthetic nitrogen (N) fertilizer has increased anthropogenic N2O emission in global croplands. Due to the advantage of biological nitrogen fixation (BNF), the introduction of legume may lessen N2O emission intensity (emission/yield) in cereal field. Yet, this speculation lacks strong evidences and the relevant mechanistic explanations. A two-year field study was conducted using static chamber method (15N isotope labeling in fertilizer) in the mono- and inter-cropping fields with soybean introduced in maize field in semiarid rainfed Loess Plateau. The results indicated that maize-soybean intercropping showed 13 % lower N2O emission intensity than monocropping, attributed to 14 % and 20 % lower annually N2O emissions in intercropped zone than that of sole maize and soybean zones respectively. Also, a 15 % higher grain yield was observed in intercropped maize relative to that in sole maize. Particularly, during the peak time of emission following N fertilization and BNF within growing season, soybean introduction reduced 25 % and 24 % of N2O emissions in intercropped zone than sole maize and soybean zone respectively. It also favored maize plant with an additional 17 % more fertilizer-derived N, 21 % less soil-derived N and 30 kg N ha−1 soybean transferred-N from BNF. The contributions of N fertilization, BNF and soil initial organic N to N2O production were lessened in intercropping. In this case, the capacity of substrates N transformation into N2O emissions was lowered. Critically, soybean introduction reduced soil ammonia oxidation (as evidenced from amoA AOA and AOB gene abundance) in intercrops’ rhizosphere, and increased soil nitrite reduction (nirS and nirK gene abundance) in maize rhizosphere, and improved soil N2O reduction to N2 (nosZ gene abundance) in soybean rhizosphere. Therefore, soybean introduction optimized N source use for lower N2O emission intensity, via reducing the transformation of substrate N into N2O, and modifying the expression of nitrification and denitrification functional genes.

Effects of biochar application methods on greenhouse gas emission and nitrogen use efficiency in paddy fields

Biochar application in rice production reduces nitrogen loss and greenhouse gases. We conducted in situ experiments for 3 years, with N210B0 (210 kg N ha−1) as the control. Two biochar application methods (B1:15 t ha−1 biochar applied once and B2: biochar applied three times at 5 t ha−1 yr−1) combined with two nitrogen levels (N210: 210 kg N ha−1 and N168: 168 kg N ha−1) were used. Soil physicochemical properties, CH4 and N2O emissions, functional gene abundance, rice yield, and nitrogen use efficiency were analyzed. Both methods improved the physicochemical properties of the soil, however, B1 was less effective than B2 in increasing soil pH, bulk density, organic carbon, total nitrogen, and microbial biomass nitrogen in year 3. B1 had a higher CH4 emission mitigation effect than B2 in 3 consecutive years, mainly due to the higher pmoA gene abundance. B1 showed a higher reduction effect of N2O emissions compared to B2 in year 1, but the opposite was observed in years 2 and 3. B2 had a higher abundance of AOB, nirK, and nosZ genes compared to B1 in year 3. Compared with N210B0, rice yields were increased by 9.1 %, 9.6 %, and 3.6 % with N210B1, N210B2, and N168B2, respectively, over 3 years, while N168B1 improved yields in the previous 2 years. Biochar improved nitrogen use efficiency over 3 consecutive years directly due to increased use efficiency of panicle fertilizer; the effect of B1 was greater than that of B2 during years 1 and 2, while the opposite was observed in year 3. Both Biochar applied once and three times appeared to be promising practices to increase yield and mitigate GHGs. From the GHGI perspective, the biochar applied once combined with 168 kg N ha−1 can further improve nitrogen use efficiency, and reduce GHGs without hindering improvements in rice yield.

Elucidating the links between N2O dynamics and changes in microbial communities following saltwater intrusions

Saltwater intrusion in estuarine ecosystems alters microbial communities as well as biogeochemical cycling processes and has become a worldwide problem. However, the impact of salinity intrusion on the dynamics of nitrous oxide (N2O) and associated microbial community are understudied. Here, we conducted field microcosms in a tidal estuary during different months (December, April and August) using dialysis bags, and microbes inside the bags encountered a change in salinity in natural setting. We then compared N2O dynamics in the microcosms with that in natural water. Regardless of incubation environment, saltwater intrusion altered the dissolved N2O depending on the initial saturation rates of N2O. While the impact of saltwater intrusion on N2O dynamics was consistent across months, the dissolved N2O was higher in summer than in winter. The N-related microbial communities following saltwater intrusion were dominated by denitrifers, with fewer nitrifiers and bacterial taxa involved in dissimilatory nitrate reduction to ammonium. While denitrification was a significant driver of N2O dynamics in the studied estuary, nitrifier-involved denitrification contributed to the additional production of N2O, evidenced by the strong associations with amoA genes and the abundance of Nitrospira. Higher N2O concentrations in the field microcosms than in natural water limited N2O consumption in the former, given the lack of an association with nosZ gene abundance. The differences in the N2O dynamics observed between the microcosms and natural water could be that the latter comprised not only indigenous microbes but also those accompanied with saltwater intrusion, and that immigrants might be functionally rich individuals and able to perform N transformation in multiple pathways. Our work provides the first quantitative assessment of in situ N2O concentrations in an estuary subjected to a saltwater intrusion. The results highlight the importance of ecosystem size and microbial connectivity in the source-sink dynamics of N2O in changing environments.

Slow-release fertilizer deep placement increased rice yield and reduced the ecological and environmental impact in Southeast China: A life-cycle perspective

An unreasonable application of nitrogen fertilizer leads to increased agricultural greenhouse gas (GHG) emissions and carbon footprint (CF). To solve this issue, slow-release fertilizer and deep placement of nitrogen fertilizer are recommended, and have been proven effective in improving nitrogen utilization efficiency and reducing nitrogen loss. However, the deep placement of fertilizer increases the cost of mechanical fuel and labor, especially in the hilly and mountainous areas of southeast China and the comprehensive effects throughout the life cycle of crop production, including economic benefits, energy flow, and environmental and ecological advantages, resulting from different application strategies for nitrogen fertilizer, are still largely unknown. In this work, a two-year field trial was conducted in Fujian Province, China, from 2021 to 2022. The main objectives of the study were to investigate the yield and GHG emissions resulting from the deep placement of slow-release fertilizer (DSK) and to assess the carbon footprint, energy use efficiency, and net economic and ecological benefits from a life-cycle perspective. The results showed that compared with deep placement of urea (DCK), slow-release fertilizer broadcasting (SK), and the traditional fertilization mode practiced by farmers (CK), the average annual yield of rice under DSK treatment significantly increased by 8.78%, 15.18%, and 23.54% respectively. Compared with CK, the contents of indole-3-acetic acid (IAA) and trans-Zeatin Riboside (t-ZR) under DSK were significantly increased and the dry matter translocation amount, Non-Structural Carbohydrates (NSC) translocation amount and Nitrogen translocation amount were significantly increased by 22.79%, 49.60% and 41.47% respectively. GHG emissions were indirectly reduced by the transport of above-ground substances to grains. The average cumulative CH4 emissions were significantly reduced by 25.34% in the DSK treatment, compared to the CK treatment, primarily as a result of decreased methanogenic gene abundance of mcra gene. Additionally, N2O emissions decreased significantly by 48.97%, compared to the CK, mainly due to a decrease in nirK gene abundance and an increase in nosZ gene abundance in denitrifying bacteria. Furthermore, the carbon footprint of the DSK treatment significantly decreased by 38.74% compared to the CK treatment, while net energy income and net ecological and economic benefits increased by 43.83% and 76.43%, respectively. Comprehensive analysis showed that deep placement of slow-release fertilizer is a sustainable nitrogen fertilizer management mode that helps to balance grain yield, environmental footprint, and economic benefits in southeast China.