Abundance Of Denitrification Genes Research Articles

Nanoparticles of Zinc Oxides Mitigated N2O Emissions in Tea Plantation Soil

The excessive application of nitrogen in tea plantations leads to severe soil acidification and N2O emission boosting. To promote sustainable agriculture, nanoparticles (NPs) have emerged as alternative fertilizers, but their effects on soil nitrification and greenhouse gas emissions in tea plantations remain unclear. In this study, the effects of NP type (ZnO-NPs and Fe2O3-NPs) and dose (0, 1, 10, and 100 mg·kg⁻1) on soil N2O emissions were investigated via a lab incubation trial. Soil pH, ammonium, and nitrate changes were also monitored during the incubation period. The abundance of functional genes related to nitrification and denitrification processes was analyzed as well. The results showed that ZnO-NPs led to a decrease in N2O emissions. The reduction effect was stronger with increasing dose and resulted in a 33% reduction at an addition rate of 100 mg·kg−1. The cumulative N2O emissions had significantly positive correlations with NH4+-N and NO3−-N. ZnO-NP addition showed a significantly negative effect on Ammonia-Oxidizing Archaea (AOA) but a positive effect on Ammonia-Oxidizing Bacteria (AOB) gene abundance. In contrast, Fe2O3-NPs showed an insignificant impact on N2O emissions and soil N content, as well as nitrification–denitrification gene abundance, regardless of different doses. These results imply that the application of ZnO-NPs may inhibit nitrification through the retarding of AOA activity. This study provided us with a potential practice to reduce N2O emissions in tea plantations by applying ZnO-NPs, but the efficiency of this reduction needs further examination under ambient conditions before field application.

Soil gross N2O emission and uptake under two contrasting agroforestry systems: riparian tree buffer versus alley-cropping tree row

AbstractIn addition to the removal of excess mineral nitrogen (N) via root uptake, trees in agroforestry systems may mitigate negative effects of high N fertilization of adjacent crops by enhancing complete denitrification of excess mineral N aside from root uptake. Presently, little is known about the potential for NO3− reduction through denitrification (conversion to greenhouse gas N2O and subsequently to non-reactive N2) in contrasting agroforestry systems: riparian tree buffer versus tree row of an upland alley-cropping system. Our study aimed to (1) quantify gross N2O emissions (both N2O + N2 emissions) and gross N2O uptake (N2O reduction to N2), and (2) determine their controlling factors. We employed the 15N2O pool dilution technique to quantify gross N2O fluxes from 0 to 5 cm (topsoil) and 40 to 60 cm (subsoil) depths with seasonal field measurements in 2019. The riparian tree buffer exhibited higher topsoil gross N2O emissions and uptake than the alley-cropping tree row (P < 0.03). Gross N2O emissions were regulated by N and carbon (C) availabilities and aeration status rather than denitrification gene abundance. Gross N2O uptake was directly linked to available C and nirK gene abundance. In the subsoil, gross N2O emission and uptake were low in both agroforestry systems, resulting from low mineral N contents possibly due to N uptake by deep tree roots. Nonetheless, the larger available C and soil moisture in the subsoil of riparian tree buffer than in alley-cropping tree row (P < 0.05) suggest its large potential for N2O uptake whenever NO3− is transported to the subsoil.

Insight into effect of polyethylene microplastic on nitrogen removal in moving bed biofilm reactor: Focusing on microbial community and species interactions

Microplastics (MPs) pollution has emerged as a global concern, and wastewater treatment plants (WWTPs) are one of the potential sources of MPs in the environment. However, the effect of polyethylene MPs (PE) on nitrogen (N) removal in moving bed biofilm reactor (MBBR) remains unclear. We hypothesized that PE would affect N removal in MBBR by influencing its microbial community. In this study, we investigated the impacts of different PE concentrations (100, 500, and 1000 μg/L) on N removal, enzyme activities, and microbial community in MBBR. Folin–phenol and anthrone colorimetric methods, oxidative stress and enzyme activity tests, and high-throughput sequencing combined with bioinformation analysis were used to decipher the potential mechanisms. The results demonstrated that 1000 μg/L PE had the greatest effect on NH4+-N and TN removal, with a decrease of 33.5 % and 35.2 %, and nitrifying and denitrifying enzyme activities were restrained by 29.5–39.6 % and 24.6–47.4 %. Polysaccharide and protein contents were enhanced by PE, except for 1000 μg/L PE, which decreased protein content by 65.4 mg/g VSS. The positive links of species interactions under 1000 μg/L PE exposure was 52.07 %, higher than under 500 μg/L (51.05 %) and 100 μg/L PE (50.35 %). Relative abundance of some metabolism pathways like carbohydrate metabolism and energy metabolism were restrained by 0.07–0.11 % and 0.27–0.4 %. Moreover, the total abundance of nitrification and denitrification genes both decreased under PE exposure. Overall, PE reduced N removal by affecting microbial community structure and species interactions, inhibiting some key metabolic pathways, and suppressing key enzyme activity and functional gene abundance. This paper provides new insights into assessing the risk of MPs to WWTPs, contributing to ensuring the health of aquatic ecosystems.

Enhancing nitrogen removal through macrophyte harvest and installation of woodchips-based floating beds in surface-flow constructed wetlands

Wetland management maintains nitrogen (N) removal capacity in mature and overgrown constructed wetlands (CWs). We evaluated whether CW management by macrophyte harvesting, and subsequent installation of woodchips-based floating beds (WFBs) planted with Glyceria maxima and Filipendula ulmaria improved N removal. In sixteen heavily overgrown experimental CWs, we applied four treatments: i) only macrophyte harvesting, ii) 5% of the harvested-CW surface covered with WFBs, iii) 20% WFBs cover, and iv) a control treatment (heavily overgrown). N removal was determined in all wetlands at nine occasions. Plant biomass accrual, N assimilation, and denitrification genes nirS, nirK, nosZI and nosZII on plant roots and woodchips from WFBs were estimated. Macrophyte harvesting improved N removal of heavily overgrown CWs, whereas subsequent WFB installation only sometimes improved N removal. Mean N removal efficiencies (± standard deviation) overall were 41 ± 15 %, 45 ± 20 %, 46 ± 16 % and 27 ± 8.3 % for treatments i to iv, respectively. Relative biomass production, root length and root surface area for G.maxima (mean ± standard deviation: 234 ± 114 %, 40 ± 6.5 cm, 6308 ± 1059 cm2g-1, respectively) were higher than those for F. ulmaria (63 ± 86 %, 28 ± 12 cm, 3131 ± 535 cm2g-1, respectively) whereas biomass N assimilation was higher for F. ulmaria (1.8 ± 0.9 gNm−2 of WFB) than for G. maxima (1.3 ± 0.5 gNm−2 of WFB). Denitrification gene abundance was higher on plant roots than on woodchips while G. maxima hosted higher root denitrification gene abundance than F. ulmaria. We conclude that macrophyte harvesting improves N removal in heavily overgrown CWs. WFBs installation has the potential to support plant growth and denitrification in surface-flow constructed wetlands. Further studies need to evaluate the long-term effects of macrophyte harvesting and WFB installation on N removal in CWs.

Insights into the abundance, expression and diversity of key denitrification genes in an ecologically managed greenhouse agricultural soil

Understanding the bacteria associated with nitrification and denitrification is crucial for comprehending the processes that lead to nitrous oxide emissions in agricultural greenhouse soils. Therefore, it is important to determine their abundance and expression to gain insight into these processes.The aim of this study was to explore the bacterial communities associated with denitrification in a greenhouse agricultural soil amended with crop residues and manure for six years. For this purpose, we proceeded to detect and quantify the genes nirK and nirS and the gene nosZ through clone library construction, sequencing, phylogenetic analysis, and quantitative polymerase chain reaction (qPCR). Sequence analysis based on the clone library revealed that many of the nirS or nirK genes detected were not closely related to known denitrifier bacteria, but some of the nosZ sequences were related to the genera such as Pseudomonas, Halomonas, and Marinobacter. Furthermore, the qPCR revealed a high abundance of DNA copies in nirK, 6.08 × 109 ± 1.16 × 109, while nirS and nosZ showed lower values, 9.05 × 106 ± 1.65 × 106 and 8.71 × 106 ± 1.44 × 106, respectively. However, the highest expression rate was observed for nirS (mRNA/DNA ratio = 3.10 × 10− 3), while nirK and nosZ showed 10-fold lower expression rates (4.4 × 10− 4 and 3.5 × 10− 4, respectively). The results of this work provide a preliminary overview of the diversity, abundance and expression of key genes associated with the denitrification process in this type of soil and are a starting point for further studies to understand how this type of soil management can influence the denitrification process.

Biodegradable film mulching increases soil microbial network complexity and decreases nitrogen-cycling gene abundance

Biodegradable plastic films have emerged as a substitute for conventional plastic films. Nevertheless, responses of plant-associated microbiomes to the application of biodegradable film mulching at field scale have received little attention. A field experiment was conducted to assess the influence of different film mulching treatments on various microbial attributes and nitrogen (N) cycling functional genes in bulk and rhizosphere soils. Biodegradable film mulching raised the bacterial Shannon index in bulk soils but not in rhizosphere soils. Biodegradable film mulching has led to an increase in the complexity and connectivity of microbial networks, as well as an enhancement in the positive association among microorganisms owing to raised soil nutrients and increased crop biomass. In biodegradable film-treated soils, both bacterial and fungal communities were primarily influenced by stochastic processes associated with dispersal limitation. Moreover, conventional plastic film mulching increased denitrification, anammox, N fixation, and dissimilatory nitrate-reduction (DNRA) gene abundance in bulk soils. In rhizosphere soils, biodegradable film mulching reduced nitrification, denitrification, anammox, N fixation, and DNRA gene abundance. Furthermore, keystone genera (e.g., Nitrosospira, Truepera, Adhaeribacter, Opitutus, and Fusarium) were affected by edaphic variables, contributing to decreased N-cycling gene abundance in biodegradable film-treated soils. Collectively, biodegradable film mulching transformed soil microbiome assembly and functional adaptation, and soil nutrient availability and plant biomass were the critical factors influencing the microbial community. These findings present a novel perspective on the diverse impacts of biodegradable and conventional film mulching on soil microbiome and N-cycling processes.

Effects of sulfamethoxazole/trimethoprim on corncob-supported solid-phase denitrification and removal performance: Microbial community, biotransformation pathways, and antibiotic resistance genes

The widespread use of antibiotics inevitably results in their discharge into the water environment from the wastewater treatment plant (WWTP) effluents. Corncob-supported solid-phase denitrification (CC-SPD) has been identified as a promising technology for nitrogen removal of WWTP effluents. The effects of sulfamethoxazole (SMX)/trimethoprim (TMP) on nitrogen removal in up-flow CC-SPD reactors and the influencing mechanisms are studied at microbial and genetic levels, and the antibiotic removal performance, degradation pathways, and antibiotic resistant genes (ARGs) are also extensively investigated. Long-term experimental results indicated that 0.05 and 2 mg/L SMX/TMP, either individually or in combination, had no inhibition on nitrogen removal performance in CC-SPD reactors. The NO3\\_\\_-N removal capacity was approximately 125.60–142.91 mg NO3\\_\\_-N/g CC. Although the microbial community structure evolved to adapt to the 0.05 mg/L SMX/TMP, the relative abundances (RAs) of denitrification genes and genes related to electron donor behaviours did not significantly alter. Furthermore, SMX, with a higher removal efficiency of 42.17 %, was more easily degradable compared to TMP, and the degradation products of SMX were more readily mineralized. Additionally, microorganisms degrade antibiotics through processes such as ring opening, hydrogenation, and bond cleavage, with the toxicity of intermediates often being lower than that of parent compounds. Moreover, 0.05 mg/L SMX/TMP did not significantly change the RAs of ARGs and movable genetic elements (MGEs). This study provides important basis for the antibiotic risk prevention in WWTP effluents and the practical application of SPD technology.

Control of nitrogen and odor emissions during chicken manure composting with a carbon-based microbial inoculant and a biotrickling filter

Although aerobic composting is usually utilized in livestock manure disposal, the emission of odorous gases from compost not only induces harm to the human body and the environment, but also causes loss of nitrogen, sulfur, and other essential elements, resulting in a decline in product quality. The impact of biotrickling filter (BTF) and insertion of carbon-based microbial agent (CBMA) on compost maturation, odor emissions, and microbial population during the chicken manure composting were assessed in the current experiment. Compared with the CK group, CBMA addition accelerated the increase in pile temperature (EG group reached maximum temperature 10 days earlier than CK group), increased compost maturation (GI showed the highest increase of 41.3% on day 14 in EG group), resulted in 36.59% and 14.60% increase in NO3−-N content and the total nitrogen retention preservation rate after composting. The deodorization effect of biotrickling filter was stable, and the removal rates of NH3, H2S, and TVOCs reached more than 90%, 96%, and 56%, respectively. Furthermore, microbial sequencing showed that CBMA effectively changed the microbial community in compost, protected the ammonia-oxidizing microorganisms, and strengthened the nitrification of the compost. In addition, the nitrifying and denitrifying bacteria were more active in the cooling period than they were in the thermophilic period. Moreover, the abundance of denitrification genes containing nirS, nirK, and nosZ in EG group was lower than that in CK group. Thus, a large amount of nitrogen was retained under the combined drive of BTF and CBMA during composting. This study made significant contributions to our understanding of how to compost livestock manure while reducing releases of odors and raising compost quality.

Interaction of reed litter and biochar presences on performances of constructed wetlands

Constructed wetlands (CWs) are frequently used for effective biological treatment of nitrogen-rich wastewater with external carbon source addition; however, these approaches often neglect the interaction between plant litter and biochar in biochar-amended CW environments. To address this, we conducted a comprehensive study to assess the impacts of single or combined addition of common reed litter and reed biochar (pyrolyzed at 300 and 500 °C) on nitrogen removal, greenhouse gas emission, dissolved organic matter (DOM) dynamics, and microbial activity. The results showed that combined addition of reed litter and biochar to CWs significantly improved nitrate and total nitrogen removal compared with biochar addition alone. Compared to those without reed litter addition, CWs with reed litter addition had more low-molecular-weight and less aromatic DOM and more protein-like fluorescent DOM, which favored the enrichment of bacteria associated with denitrification. The improved nitrogen removal could be attributed to increases in denitrifying microbes and the relative abundance of functional denitrification genes with litter addition. Moreover, the combined addition of reed litter and 300 °C-heated biochar significantly decreased nitrous oxide (30.7 %) and methane (43.9 %) compared to reed litter addition alone, while the combined addition of reed litter and 500 °C-heated biochar did not. This study demonstrated that the presences of reed litter and biochar in CWs could achieve both high microbial nitrogen removal and relatively low greenhouse gas emissions.

Synergistic denitrification mechanism of domesticated aerobic denitrifying bacteria in low-temperature municipal wastewater treatment

To address the problems of low efficacy and low microbial activity in low-temperature municipal wastewater treatment, this study utilized an air-lift micro-pressure internal circulation integrated reactor (AMICIR). Through controlling the amount of aeration and dissolved oxygen (DO) in the reactor, AMICIR creates alternating aerobic and anaerobic environments, explores the enrichment conditions of aerobic denitrifying bacteria, examines the changes in pollutant removal efficiency and the characteristics of bacterial colony structure during the process of enrichment of aerobic denitrifying bacteria in the system, and reveals the mechanism of nitrogen removal by aerobic denitrifying bacteria cooperating with anaerobic denitrifying bacteria in the low-temperature municipal wastewater treatment system. Experimental results showed average removal rates of NH4+-N, chemical oxygen demand (COD), total phosphorus (TP), and total nitrogen (TN) reaching 93.85%, 89.30%, 92.75%, and 75.4%, respectively. The microorganisms secreted large amounts of proteins and polysaccharides, forming zoogloea and anaerobic microenvironments conducive to traditional denitrification reactions. IlluminaMiSeq sequencing analysis revealed the presence of anaerobic phyla. The system was enriched with a large number of microorganisms, and aerobic denitrifying bacteria (Flavobacterium, Rhodoferax, and Pseudomonas) were successfully cultured. Flavobacterium emerged as the dominant species, with relative abundance ranging from 18.56% to 22.60%. Functional gene prediction indicated high abundance of aerobic denitrification genes, such as napA. Aerobic denitrifying bacteria were successfully enriched in the system to improve nitrogen removal from municipal wastewater at low temperatures.

Denitrification regulates spatiotemporal pattern of N2O emission in an interconnected urban river-lake network

Urban rivers are hotspots of N2O production and emission. Interconnected river-lake networks are constructed to improve the water quality and hydrodynamic conditions of urban rivers in many cities of China. However, the impact of the river-lake connectivity project on N2O production and emission remains unclear. This study investigated dissolved N2O and emission of the river-lake network in Wuhan City, China from March 2021 to December 2021. The results showed that river-lake connection greatly decreased riverine Nitrogen (N) concentration and increased dissolved oxygen (DO) concentration compare to traditional urban rivers. N2O emissions from the urban river interconnected with lakes (LUR: 67.3 ± 92.6 μmol/m2/d) were much lower than those from the traditional urban rivers (UR: 467.3 ± 1075.7 μmol/m2/d) and agricultural rivers (AR: 20.4 ± 15.3μmol/m2/d). Regression tree analysis suggested that the N2O concentrations were extremely high when hypoxia exists (DO < 1.6 mg/L), and TDN was the primary factor regulating N2O concentrations when hypoxia does not occur. Thus, we ascribe the low N2O emission in the LUR and AR to the lower N contents and higher DO concentrations. The microbial process of N2O production and consumption were quantitatively estimated by isotopic models. The mean proportion of denitrification derived N2O (fbD) was 63.5 %, 55.6 %, 42.3 % and 42.7 % in the UR, LUR, lakes and AR, suggested denitrification dominated N2O production in the urban rivers, but nitrification dominated N2O production in the lakes and AR. The positive correlation between logN2O and fbD suggested that denitrification is the key process to regulate the N2O production and emission. The abundance of denitrification genes (nirS and nirK) was much higher than that of nitrification genes (amoA and amoB), also evidenced that denitrification was the main N2O source. Therefore, river-lake interconnected projects changed the nutrients level and hypoxic condition, leading to the inhibition of denitrification and nitrification, and ultimately resulting in a decrease of N2O production and emission. These results advance the knowledge on the microbial processes that regulate N2O emissions in inland waters and illustrate the integrated management of water quality and N2O emission.

Differential Nitrous oxide emission and microbiota succession in constructed wetlands induced by nitrogen forms

Nitrous oxide (N2O) emission during the sewage treatment process is a serious environmental issue that requires attention. However, the N2O emission in constructed wetlands (CWs) as affected by different nitrogen forms in influents remain largely unknown. This study investigated the N2O emission profiles driven by microorganisms in CWs when exposed to two typical nitrogen sources (NH4+-N or NO3–-N) along with different carbon source supply (COD/N ratios: 3, 6, and 9). The results showed that CWs receiving NO3–-N caused a slight increase in total nitrogen removal (by up to 11.8 %). This increase was accomplished by an enrichment of key bacteria groups, including denitrifiers, dissimilatory nitrate reducers, and assimilatory nitrate reducers, which enhanced the stability of microbial interaction. Additionally, it led to a greater abundance of denitrification genes (e.g., nirK, norB, norC, and nosZ) as inferred from the database. Consequently, this led to a gradual increase in N2O emission from 66.51 to 486.77 ug-N/(m2·h) as the COD/N ratio increased in CWs. Conversely, in CWs receiving NH4+-N, an increasing influent COD/N ratio had a negative impact on nitrogen biotransformation. This resulted in fluctuating trend of N2O emissions, which decreased initially, followed by an increase at later stage (with values of 122.87, 44.00, and 148.59 ug-N/(m2·h)). Furthermore, NH4+-N in the aquatic improved the nitrogen uptake by plants and promoted the production of more root exudates. As a result, it adjusted the nitrogen-transforming function, ultimately reducing N2O emissions in CWs. This study highlights the divergence in microbiota succession and nitrogen transformation in CWs induced by nitrogen form and COD/N ratio, contributing to a better understanding of the microbial mechanisms of N2O emission in CWs with NH4+-N or NO3–-N at different COD/N ratios.

Metagenomic analysis reveals the microbial response to petroleum contamination in oilfield soils

The response of the microbes to total petroleum hydrocarbons (TPHs) in three types of oilfield soils was researched using metagenomic analysis. The ranges of TPH concentrations in the grassland, abandoned well, working well soils were 1.16 × 102–3.50 × 102 mg/kg, 1.14 × 103–1.62 × 104 mg/kg, and 5.57 × 103–3.33 × 104 mg/kg, respectively. The highest concentration of n-alkanes and 16 PAHs were found in the working well soil of Shengli (SL) oilfield compared with those in Nanyang (NY) and Yanchang (YC) oilfields. The abandoned well soils showed a greater extent of petroleum biodegradation than the grassland and working well soils. Α-diversity indexes based on metagenomic taxonomy showed higher microbial diversity in grassland soils, whereas petroleum-degrading microbes Actinobacteria and Proteobacteria were more abundant in working and abandoned well soils. RDA demonstrated that low moisture content (MOI) in YC oilfield inhibited the accumulation of the petroleum-degrading microbes. Synergistic networks of functional genes and Spearman's correlation analysis showed that heavy petroleum contamination (over 2.10 × 104 mg/kg) negatively correlated with the abundance of the nitrogen fixation genes nifHK, however, in grassland soils, low petroleum content facilitated the accumulation of nitrogen fixation genes. A positive correlation was observed between the abundance of petroleum-degrading genes and denitrification genes (bphAa vs. nirD, todC vs. nirS, and nahB vs. nosZ), whereas a negative correlation was observed between alkB (alkane- degrading genes) and amo (ammonia oxidation), hao (nitrification). The ecotoxicity of petroleum contamination, coupled with petroleum hydrocarbons (PH) degradation competing with nitrifiers for ammonia inhibited ammonia oxidation and nitrification, whereas PH metabolism promoted the denitrification process. Moreover, positive correlations were observed between the abundance of amo gene and MOI, as well as between the abundance of the dissimilatory nitrate reduction gene nirA and clay content. Thus, improving the soil physicochemical properties is a promising approach for decreasing nitrogen loss and alleviating petroleum contamination in oilfield soils.

Induced and natural moss soil crusts accelerate the C, N, and P cycles of Pb[sbnd]Zn tailings

Nutrient deficiency is the primary obstacle in tailing ecological restoration besides high heavy metal content. Biological soil crusts (BSCs) are known for their C and N fixation capabilities and play a crucial role in soil P cycle. BSCs are widespread in tailings and provide a potential ecological restoration approach. In 2022, we carried out an on-site restoration on a PbZn tailing pond in Yunnan Province, China. BSCs were propagated by natural moss crust fragment inoculation. The induced moss crusts (IMCs) were monitored at 0, 45, 90, and 135 days and compared with natural moss crusts (NMCs). The chlorophyll-a content and abundance of biotic organisms increased over time, reaching a peak at 135 days and surpassing that of NMCs. Moss crusts increased the content of C, N, and P nutrients and enzyme activities in the 0.5 cm surface soil. They also reduced the DTPA-extractable Pb content. Moss crusts significantly increased the content of fulvic/humic and protein-like/polyphenol substances, thereby raising the humic index of soil dissolved organic matter (especially NMCs). Furthermore, moss crusts also raised the abundance of nitrification (AOB and Nsr), denitrification (narG, napA, qnorB, and nosZ), and P-cycling (gcd, appA, phoC, phoA, and phoD) genes, especially IMCs after a 135-day inoculation. NMCs exhibited higher moss abundance measured via eukaryotic photoautotrophs. Moss crusts increased photosynthetic bacteria abundance (e.g., Leptolyngbya and Nostocales) and reduced the dominance of chemoautotrophic bacteria, especially the dark sulfide oxidation bacteria (Betaproteobacteriales). This trend was more pronounced in NMCs. Overall, IMCs can recover the functions of NMCs, and in some cases (e.g., abundance and diversity of biotic community, soil nutrient and N & P cycle genes), even surpass them. Our research provides new insights into the tailing ecological restoration.

Impacts of biostimulation and bioaugmentation on woodchip bioreactor microbiomes.

Woodchip bioreactors (WBRs) are used to remove nutrients, especially nitrate, from subsurface drainage. The nitrogen removal efficiency of WBRs, however, is limited by low temperatures and the availability of labile carbon. Bioaugmentation and biostimulation are potential approaches to enhance nitrate removal of WBRs under cold conditions, but their effectiveness is still unclear. Here, we clarified the effects of bioaugmentation and biostimulation on the microbiomes and nitrate removal rates of WBRs. As a bioaugmentation treatment, we inoculated WBR-borne cold-adapted denitrifying bacteria Cellulomonas cellasea strain WB94 and Microvirgula aerodenitrificans strain BE2.4 into the WBRs located at Willmar, MN, USA. As a biostimulation treatment, acetate was added to the WBRs to promote denitrification. Woodchip samples were collected from multiple locations in each WBR before and after the treatments and used for the microbiome analysis. The 16S rRNA gene amplicon sequencing showed that the microbiomes changed by the treatments and season. The high-throughput quantitative PCR for nitrogen cycle genes revealed a higher abundance of denitrification genes at locations closer to the WBR inlet, suggesting that denitrifiers are unevenly present in WBRs. In addition, a positive relationship was identified between the abundance of M. aerodenitrificans strain BE2.4 and those of norB and nosZ in the WBRs. Based on generalized linear modeling, the abundance of norB and nosZ was shown to be useful in predicting the nitrate removal rate of WBRs. Taken together, these results suggest that the bioaugmentation and biostimulation treatments can influence denitrifier populations, thereby influencing the nitrate removal of WBRs. IMPORTANCE Nitrate pollution is a serious problem in agricultural areas in the U.S. Midwest and other parts of the world. Woodchip bioreactor is a promising technology that uses microbial denitrification to remove nitrate from agricultural subsurface drainage, although the reactor's nitrate removal performance is limited under cold conditions. This study showed that the inoculation of cold-adapted denitrifiers (i.e., bioaugmentation) and the addition of labile carbon (i.e., biostimulation) can influence the microbial populations and enhance the reactor's performance under cold conditions. This finding will help establish a strategy to mitigate nitrate pollution.

Microplastics enhance nitrogen loss from a black paddy soil by shifting nitrate reduction from DNRA to denitrification and Anammox

Microplastics (MPs) are frequently detected emerging pollutants in soil that can endanger farmland ecosystems; however, little is known about their impacts on dissimilatory nitrate reduction processes in paddy soil. Here, using the 15N-tracer and microbial molecular techniques, we investigated the effects of MPs (200–400 μm) made of polystyrene (PS), polyvinyl chloride (PVC), and polyethylene (PE) on denitrification, anaerobic ammonium oxidation (Anammox), and dissimilatory nitrate reduction to ammonium (DNRA) and the associated microbial community in a black paddy soil. All MPs increased the Anammox rate by 6.6 %–745 % and decreased the DNRA rate by 15.1 %–74.2 %, while MPs of PS and PE significantly increased the denitrification rate by 79.3 %–102.3 % and 34.8 %–62.1 %, respectively. The MPs promoted the partitioning of NO3− towards denitrification and Anammox while inhibiting DNRA, as suggested by the decreased relative contributions of DNRA from 24.1 % to 5.4 %–14.2 % following MPs amendment. This was attributed to the increased denitrification gene abundance and the enriched specific denitrifier taxa, as well as the decreased DNRA gene abundance. Our findings suggest that the stimulated denitrification and Anammox by MPs, accompanied by the suppression of DNRA, may lead to substantial nitrogen loss in paddy fields, underscoring the need to further evaluate the environmental behaviors of MPs in agricultural ecosystems.

Nitrification and Denitrification Gene Abundances under Stable Soil Chemical Properties Established by Long-Term Compost Fertilization

Rice paddies are dynamic areas for microbe-mediated nitrogen cycling and this could be driven by the long-term alteration of paddy soil edaphic factors. The objective of this study is to evaluate the lasting impact of long-term compost fertilization on the soil chemical properties of paddy fields, determining the size of the microbial guilds involved in nitrification and denitrification. Soil sampling was done on paddy fields without fertilizer, with NPK + compost, and with compost application, and the soil chemical properties of the fields were determined from 2018 to 2021. The abundance of genes related to nitrogen cycling was measured using quantitative PCR (qPCR). Annual analyses showed that the studied soils have attained stable, baseline chemical properties with significantly increased phosphorus (P2O5), potassium (K), SOM, and total nitrogen (TN) due to decades of fertilization with NPK + compost and compost. Consequently, the genes related to bacterial amoA, nosZI, and nosZII were significantly increased in Com- and NPKCom-amended soils compared to the NF paddy field. The nirK gene abundance was unaffected after long-term amendment with compost. A positive correlation was found between the archaeal amoA gene abundance and DOC, while SOM and TN were consistently positively correlated with the abundances of bacterial amoA, nosZI, and nosZII genes, in addition to interactions with potassium and DOC. Principal component analysis (PCA) indicated soil variabilities across treatments, where the unfertilized paddy field contained lower SOM and nutrient contents with a characteristic nirK gene abundance. Similar variabilities in terms of the SOM, TN, K, nosZI, and nosZII gene abundances were observed in the Com and NPKCom paddy fields. Long-term amendment with NPK + compost and compost created soil paddy fields with stable soil chemical properties with higher SOM and nutrient contents, which established higher abundances of genes associated with denitrification and nitrification that were observed during the fallow period.

The effect of antecedent soil moisture conditions on soil nitrous oxide and dinitrogen dynamics after wetting: An intact soil core study

Ruminant excreta urinary‑nitrogen (N) deposited onto pasture soil generates inorganic-N concentrations in excess of the pasture's immediate requirements. Consequently, nitrous oxide (N2O) and dinitrogen (N2) may be emitted from the urine-affected soil via denitrification. Soil moisture plays a key role in determining the end-product of denitrification. Although the effects of soil moisture and drying-wetting cycles on soil N2O production have been widely studied using sieved soils in a controlled environment, these experiments may not reflect the actual conditions in the field due to the lack of plant cover and the disturbed soil structure. To address this an experiment was performed using intact soil cores (7.5 cm depth), with pasture plants present, under controlled environmental conditions (20 °C, 8 h of light per day). The 15N flux method was used to investigate how antecedent soil moisture affected soil N2O production and consumption in a pasture soil after wetting with a nitrate (NO3−) solution. The results showed that the antecedent dry condition (25 % WFPS, Dp/Do = 0.1783) increased the magnitude of the N2O flux peak after the soil was wetted (75 % WFPS, Dp/Do = 0.006) when compared to antecedent wet conditions (50 % WFPS, Dp/Do = 0.0503). There were no differences in the denitrification and nitrification related functional gene abundances between the two antecedent conditions post-wetting, indicating that the effect was not regulated by microbial changes but by the intensity of the “Birch effect” induced by soil wetting. The N2O reduction related gene (nosZII) abundance decreased after wetting with the NO3− solution, indicating a suppression effect on N2O consumption from the NO3− applied that overrode the potential “Birch effect” on N2 emission. The results of the study indicate that rewetting of antecedent dry soil increase N2O emissions but not N2 emissions when the soil NO3− concentration is high.

New insights into the effects of wetland plants on nitrogen removal pathways in constructed wetlands with low C/N ratio wastewater: Contribution of partial denitrification-anammox.

Nitrogen (N) removal in constructed wetlands (CWs) was often challenged by limited denitrification due to the lack of carbon source, and wetland plants would be more important in carbon (C) and N cycling in CWs with influent of low carbon to nitrogen (C/N) ratio. In this study, the underlying mechanisms of nitrate nitrogen (NO3--N) removal under different low C/N ratios were revealed by constructing microcosm CWs, and the unplanted group was set as the control to explore the role of plants in N removal. The results showed that plants and the concentration of influent carbon significantly affected NO3--N and total nitrogen (TN) removal (p < 0.05). The presence of plants significantly increased the concentration of DO and wetland plant-derived DOM (p < 0.05). The enhanced NO3--N and TN removal with increased C/N ratio attributed to high denitrification activity reflected in the abundance of denitrification microbes and genes. However, the contribution of partial denitrification-anammox (PDN/AMX) to N removal in CWs decreased from more than 75.3% at the C/N ratio of 0 to 70.4% and 22.3% with the C/N ratio increased to 1.5 and 3, respectively. Furthermore, the PDN/AMX process was negatively correlated with favorable oxygen environment in the planted group and plants roots carbon secretion, but the overall N removal efficiency of the CWs was enhanced by increased abundance of N removal-related functional genes in the presence of plants. Abovementioned results provided new insights to explain the mechanism of N removal in CWs under low C/N ratio.

Diluted pyroligneous vinegar promoted Rhododendron growth by changing functional genes involved in N cycling in the rhizosphere

Excessive chemical fertilization in protected horticulture has caused nutrient accumulation, reduction of soil biodiversity, and subsequently soil degradation. Pyroligneous vinegar is a byproduct of pyrolysis for biochar production. While there has been an increase in interest in using pyroligneous vinegar as an amendment to improve soil quality, a paucity of data exists of its effect on the growth of acidophilous plants such as azalea (Rhododendron). To address this, we investigated the changes in the growth of azalea and soil microbial abundance, specifically those involved in nitrogen cycling in the rhizosphere with amendment of conventional inorganic NPK fertilizer (NPK) and four dilutions of pyroligneous vinegar each with matched NPK content. The results showed that the treatments of pyroligneous vinegar increased plant biomass (51%-99%), root length (29%-81%) and surface area (63%-151%), and nitrogen use efficiency (83%-299%) compared to NPK alone. The pyroligneous vinegar treatments decreased soil pH and increased the contents of rhizosphere soil dissolved organic carbon and nitrate, and changed potential denitrification activity. Compared with NPK, pyroligneous vinegar increased archaeal and bacterial amoA genes (AOA and AOB) abundances (45% − 79% and 39% − 75%, respectively) in nitrification, but inhibited the denitrification gene abundances (i.e. nirS and nirK abundance up to 21% lower). Pyroligneous vinegar increased the ratio of nosZ/(nirS + nirK), suggesting an increased potential to lower nitrous oxide emissions.Correlation analysis indicated that nitrogen use efficiency correlated positively with AOA and AOB, but negatively with nirS and nirK abundance. Therefore, our study suggests that pyroligneous vinegar could improve the growth of acidophilous plants and increase their nitrogen use efficiency by changing nitrogen cycling related functional genes, and could be a potential soil amendment in protected horticulture.