Denitrifying Activity Research Articles

Bio-augmentation with dissimilatory nitrate reduction to ammonium (DNRA) driven sulfide-oxidizing bacteria enhances the durability of nitrate-mediated souring control

Biological souring (producing sulfide) is a global challenge facing anaerobic water bodies, especially the oil reservoir fluids. Nitrate injection has demonstrated great potential in souring control, and dissimilatory nitrate reduction to ammonium (DNRA) bacteria was proposed to play crucial roles in the process. How to durably control souring with nitrate amendment, however, remains undiscovered. Herein, Gordonia sp. TD-4, a DNRA-driven sulfide-oxidizing bacterium, was used to elucidate the effects of bio-augmentation with DNRA bacteria on the durability of nitrate-mediated souring control. The results revealed that nitrate amendment combined with bio-augmentation with TD-4 after souring could effectively control souring and enhance the durability of nitrate-mediated souring control, while nitrate amendment before souring failed to persistently control souring. Nitrate amendment before and after souring resulted in different evolution dynamics of nitrate-reducing bacteria. Denitrifying bacteria were enriched in reactors amended with nitrate before souring or in dissolved sulfide exhausted reactors amended with nitrate after souring. The heterotrophic denitrifying activity of denitrifying bacteria, however, decreased the durability of nitrate-mediated souring control. Comparative and functional genomics analysis identified potential niche adaptation mechanisms (autotrophic and heterotrophic nitrate/nitrite reduction, including DNRA and denitrification) of predominant SRB in nitrate-amended environments, which were responsible for the rapid resumption of sulfide accumulation after the depletion of nitrate and nitrite. Pulsed injection of nitrate combined with bio-augmentation with DNRA-driven sulfide-oxidizing bacteria was proposed as a potential method to enhance the durability of nitrate-mediated souring control. The findings were innovatively applied to simultaneous bio-demulsification and souring control of emulsified and sour produced water from the petroleum industry.

Spring barley performance benefits from simultaneous shallow straw incorporation and top dressing as revealed by rhizotrons with resealable sampling ports

Plant residues with larger carbon (C) to nitrogen (N) ratios can stimulate microbial growth and thereby protect soil nutrients from leaching. In poorly fertilized soil, excessive immobilization may limit nutrient availability and thus plant growth. Little is known about the impact of a shallow straw incorporation on soil microbial regulation of top-dressing fertilizer nutrients and spring crop establishment. We aimed to evaluate if wheat straw in combination with mineral fertilizer has more positive effects on plant performance than mineral fertilization alone and if this relates to changes of the extractable C:N:P ratio and microbial activity close to the roots. In order to conduct small-scale sampling with minimal disturbance during growth of spring barley (Hordeum vulgare L.), we developed rhizotrons with resealable ports. Rhizotrons were filled with loamy-sandy soil and fertilized with an equivalent of 150 kg N and 80 kg P ha−1. Half of the rhizotrons received the top dressing together with 4500 kg wheat straw-C ha−1. Throughout a 90-day greenhouse experiment, we analyzed soil C:N:P dynamics, and carbon dioxide (CO2) and nitrous oxide (N2O) emission, together with microbial biomass, selected bacterial genes (abundance), and transcripts (activity) in bulk and root-affected soil at multiple times. We focused on nitrifiers and denitrifiers and linked our data to barley growth. Interactions between straw and roots caused shifts towards larger C:P and C:N ratios in root-affected soil. These shifts were associated with increased 16S rRNA transcripts and denitrifier activities. Straw increased microbial biomass by 124% in the topsoil and at the same time increased root biomass by 125% and number of tillers by 80%. We concluded that microbial activation at the root-straw interface may positively feed back on soil nutrient regulation and plant performance. Further research has to evaluate if plant roots actively prime mining of previously immobilized nutrients in the straw detritusphere or if effects of pathogen suppression and growth promotion are dominating.

Organic matter removal in a simultaneous nitrification–denitrification process using fixed-film system

Swine wastewater treatment is a complex challenge, due to the high organic matter (OM) and nitrogen (N) concentrations which require an efficient process. This study focused on evaluating two different support media for OM and N removal from an Upflow Anaerobic Sludge Blanket (UASB) reactor fed with swine wastewater. Maximum specific nitrification (MSNA) and denitrification (MSDA) activity test for both biofilm and suspended biomass were carried out using as supports: polyurethane foam (R1) and polyethylene rings (R2). The results showed that R2 system was more efficiently than R1, reaching OM removal of 77 ± 8% and N of 98 ± 4%, attributed to higher specific denitrifying activity recorded (5.3 ± 0.34 g NO3-N/g TVS∙h). Furthermore, 40 ± 5% of the initial N in the wastewater could have been transformed into molecular nitrogen through SND, of which only 10 ± 1% was volatilized. In this sense, MSDA tests indicated that suspended biomass was responsible for at least 70% of N removal and only 20% can be attributed to biofilm. SND could be confirmed with the analysis of microbial diversity, due to the presence of the genus Pseudomonas dominated the prokaryotic community of the system in 54.4%.

Physiological and kinetic evaluation of ampicillin oxidation as unique electron source by a denitrifying sludge

AbstractBACKGROUNDDenitrification allows the simultaneous oxidation of recalcitrant compounds and the reduction of nitrate. Scarce information on consumption and oxidation of ampicillin (AMP) as sole electron source is found in the literature. The physiological and kinetic ability of a denitrifying sludge without previous exposure to antibiotics for oxidizing several AMP concentrations was evaluated.RESULTSComplete denitrification was accomplished in batch reference assays with acetate (20–100 mg C L−1) and nitrate (C/N 1.1) within 4 h of culture, resulting in yields of HCO3− (YHCO3−) and N2 (YN2) of 0.89–1.0 mg C or N produced mg−1 C or N consumed, respectively. Assays amended with different concentrations of AMP (5–100 mg C L−1) as unique electron source at stoichiometric C/N ratio (0.86) showed that within 8–12 h of culture the antibiotic was totally consumed and oxidized to HCO3− and CO2 (YHCO3− between 0.27 and 0.67 and YCO2 between 0.35 and 0.54 mg C produced mg−1 C consumed), while nitrate was mostly reduced to N2 (YN2 = 0.77–1.0 mg N produced mg−1 N consumed). At AMP concentrations higher than 20 mg C L−1, N2O and carbon intermediates production was detected. Decreases of 20–90% in denitrifying specific rates were determined regarding the denitrifying reference. After exposure to the antibiotic, the sludge was able to recover its denitrifying activity in the absence of AMP with no N2O accumulation, whereas only 39% of the denitrification rates were recuperated.CONCLUSIONIt was clearly evidenced that AMP was used as electron donor for conducting denitrification; however, the denitrifying process occurred at lower specific rates. © 2021 Society of Chemical Industry (SCI).

Comparison of Compaction Alleviation Methods on Soil Health and Greenhouse Gas Emissions

Soil compaction can occur due to trafficking by heavy equipment and be exacerbated by unfavourable conditions such as wet weather. Compaction can restrict crop growth and increase waterlogging, which can increase the production of the greenhouse gas nitrous oxide. Cultivation can be used to alleviate compaction, but this can have negative impacts on earthworm abundance and increase the production of the greenhouse gas carbon dioxide. In this study, a field was purposefully compacted using trafficking, then in a replicated plot experiment, ploughing, low disturbance subsoiling and the application of a mycorrhizal inoculant were compared as methods of compaction alleviation, over two years of cropping. These methods were compared in terms of bulk density, penetration resistance, crop yield, greenhouse gas emissions and earthworm abundance. Ploughing alleviated topsoil compaction, as measured by bulk density and penetrometer resistance, and increased the crop biomass in one year of the study, although no yield differences were seen. Earthworm abundance was reduced in both years in the cultivated plots, and carbon dioxide flux increased significantly, although this was not significant in summer months. Outside of the summer months, nitrous oxide production increased in the non-cultivated treatments, which was attributed to increased denitrifying activity under compacted conditions.

Formation of links between microorganisms of certain groups in haplic luvisol under the influence of mineral fertilization and liming

The aim of the research was to form connections between separate groups of microorganisms as a part of groups of gray forest soil (Haplic Luvisol) during the cultivation of spring wheat with the use of various fertilizers and liming systems. Research methods: microbiological, laboratory-analytical, statistical. The research was carried out in the landfill monitoring system on the basis of a stationary experiment of the Department of Agrosoil Science and Soil Microbiology at the NSC “Institute of Agriculture of NAAS” - Development and improvement of intensive cultivation technologies of agricultural crops on the basis of the expanded soil fertility reproduction. The main results of the study: The number of ammonifiers is positively correlated with the total number of microorganisms (0,955), the number of mineral nitrogen immobilizers, denitrifiers, nitrifiers, cellulose-destroying bacteria, actin- and micromycetes, the coefficient of nitrogen mineralization compounds, the total biological activity (0,911). The conclusions of previous research are confirmed: Azotobacter is not a diagnostic microorganism for effective soil fertility, as evidenced by the insignificance of the correlation coefficients between the number of Azotobacter and the yield of spring wheat (0,265), winter wheat (0,131), soybeans (0,303). The number of Azotobacter correlates inversely with most of the studied indicators, especially significant - with the number of polysaccharide-synthesizing microorganisms, actinomycetes, melanin-synthesizing micromycetes (-0,719), acid-forming microorganisms (- 0,611), physiological and biochemical activity of own cells, pedotrophic index. The direct nature of the relationship is established between the number of polysaccharide-synthesizing microorganisms as a diagnostic group for the optimal mineral nutrition of plants and the number of mineral nitrogen immobilizers (0,854), pedotrophs (0,970), cellulose-destructive (0,724) and autochthonous (0,878) microorganisms, actinomycetes, micromycetes, the total number of microorganisms (0,588), probability of colony formation of denitrifiers, autochthonous microorganisms and micromycetes, nitrogen mineralization coefficient, total biological activity (0,646). Indigenous microorganisms show a high level of direct dependence on the number of mineral nitrogen immobilizers, pedotrophs, cellulose-destroying and polysaccharide-synthesizing microorganisms, the total number of microorganisms, physiological and biochemical activity of denitrifiers and own cells (0,935).

Revealing the effects of static magnetic field on the anoxic/oxic sequencing batch reactor from the perspective of electron transport and microbial community shifts

The effects of static magnetic field (SMF) on an anoxic/oxic sequencing batch reactor were investigated from the perspective of electron transport via determining the variations of reduced/oxidized nicotinamide adenine dinucleotide (NADH/NAD+) ratio, NADH concentration, electron transport system activity (ETSA), poly-β-hydroxybutyrate (PHB), extracellular polymeric substances (EPS), as well as the gene expression under different conditions. Moreover, the shifts of microbial community were also analyzed. The application of SMF with an appropriate intensity significantly improved the performance of the process, the abundance of the anoxic denitrifiers, and the activity of the aerobic denitrifiers. The NADH content, as well as ETSA were also enhanced, therefore, the total nitrogen removal efficiency of the process was increased. However, the overhigh SMF intensity resulted in the change of microbial community, meanwhile, had negative effects on the metabolism of microorganisms. Selecting a proper intensity is crucial for the SMF-enhanced biological wastewater treatment process.

Effect of the Oxidative Phosphorylation Uncoupler Para-Nitrophenol on the Activated Sludge Community Structure and Performance of a Submerged Membrane Bioreactor

In this work, the metabolic uncoupler para-nitrophenol (pNP) was applied to suppress excess sludge production and to investigate its effects on the system’s performance and activated sludge community structure. The COD removal efficiency decreased from 99.0% to 89.5% prior to and after pNP addition, respectively. Application of pNP transiently reduced NH4+-N, NO3−-N and NO2−-N removal efficiencies, suggesting partial inhibition of both nitrifying and denitrifying activity. However, no changes in the relative abundance of the nitrifying bacteria occurred. Phosphorus removal efficiency was sharply reduced after pNP addition, as the consequence of hydrolysis of stored cell reserves. Tetrasphaera, a key polyphosphate accumulating organism, was also affected by the addition of pNP, a fact that highly influenced system’s ability to remove phosphorus. A drastic drop in Soluble Microbial Products (SMP) and Extracellular Polymeric Substances (EPS) was also detected shortly after the introduction of the uncoupler. On the other hand, MBR’s physicochemical parameters were restored to initial values a week after the addition of pNP. Moreover, remarkable changes in beta-diversity were noted after pNP addition. An increase of Bacteroidetes, Gammaproteobacteria and Firmicutes over Actinobacteria and Alphaproteobacteria was also observed after pNP addition.

Seasonal changes in the abundance and activity of bacterial and fungal denitrifying communities associated with different compost amendments.

Composts can be efficient organic amendments in potato culture as they can supply carbon and nutrients to the soil. However, more information is required on the effects of composts on denitrification and nitrous oxide emissions (N2O) and emission-producing denitrifying communities. The effects of three compost amendments (municipal source separated organic waste compost (SSOC), forestry waste mixed with poultry manure compost (FPMC), and forestry residues compost (FRC)) on fungal and bacterial denitrifying communities and activity was examined in an agricultural field cropped to potatoes during the fall, spring, and summer seasons. The denitrification enzyme activity (DEA), N2O emissions, and respiration were measured in parallel. N2O emission rates were greater in FRC-amended soils in the fall and summer, whereas soil respiration was highest in the SSOC-amended soil in the fall. A large number of nirK denitrifying fungal transcripts were detected in the fall, coinciding with compost application, while the greatest nirK bacterial transcripts were measured in the summer when plants were actively growing. Denitrifying community and transcript levels were poor predictors of DEA, N2O emissions, or respiration rates in compost-amended soil. Overall, the sampling date was driving the population and activity levels of the three denitrifying communities under study.

Co-culturing Hyphomicrobium nitrativorans strain NL23 and Methylophaga nitratireducenticrescens strain JAM1 allows sustainable denitrifying activities under marine conditions.

BackgroundHyphomicrobium nitrativorans strain NL23 and Methylophaga nitratireducenticrescens strain JAM1 are the principal bacteria involved in the denitrifying activities of a methanol-fed, fluidized-bed marine denitrification system. Strain NL23 possesses the complete denitrification pathway, but cannot grow under marine conditions in pure cultures. Strain JAM1 is a marine bacterium that lacks genes encoding a dissimilatory nitrite (NO2−) reductase and therefore cannot reduce NO2−. Here, we report the characterization of some of their physiological traits that could influence their co-habitation. We also perform co-cultures to assess the potential synergy between the two strains under marine and denitrifying conditions.MethodologyAnoxic planktonic pure cultures of both strains were grown with different concentrations of nitrate (NO3−). Anoxic planktonic co-cultures could only be cultured on low NaCl concentrations for strain NL23 to grow. Biofilm co-cultures were achieved in a 500-mL bioreactor, and operated under denitrifying conditions with increasing concentrations of NaCl. NO3− and NO2− concentrations and the protein content were measured to derive the denitrification rates. The concentrations of both strains in co-cultures were determined by quantitative PCR (qPCR). Ectoine concentration was measured by mass spectrometry in the biofilm co-culture. The biofilm was visualized by fluorescence in situ hybridization. Reverse-transcription-qPCR and RNA-seq approaches were used to assess changes in the expression profiles of genes involved in the nitrogen pathways in the biofilm cultures.ResultsPlanktonic pure cultures of strain JAM1 had a readiness to reduce NO3− with no lag phase for growth in contrast to pure cultures of strain NL23, which had a 2-3 days lag phase before NO3− starts to be consumed and growth to occur. Compared to strain NL23, strain JAM1 has a higher µmax for growth and higher specific NO3− reduction rates. Denitrification rates were twice higher in the planktonic co-cultures than those measured in strain NL23 pure cultures. The biofilm co-cultures showed sustained denitrifying activities and surface colonization by both strains under marine conditions. Increase in ectoine concentrations was observed in the biofilm co-culture with the increase of NaCl concentrations. Changes in the relative transcript levels were observed in the biofilm culture with genes encoding NapA and NapGH in strain NL23. The type of medium had a great impact on the expression of genes involved in the N-assimilation pathways in both strains.ConclusionsThese results illustrate the capacity of both strains to act together in performing sustainable denitrifying activities under marine conditions. Although strain JAM1 did not contribute in better specific denitrifying activities in the biofilm co-cultures, its presence helped strain NL23 to acclimate to medium with NaCl concentrations >1.0%.

Copper nitrite reductase from Sinorhizobium meliloti 2011: Crystal structure and interaction with the physiological versus a nonmetabolically related cupredoxin-like mediator.

We report the crystal structure of the copper-containing nitrite reductase (NirK) from the Gram-negative bacterium Sinorhizobium meliloti 2011 (Sm), together with complex structural alignment and docking studies with both non-cognate and the physiologically related pseudoazurins, SmPaz1 and SmPaz2, respectively. S. meliloti is a rhizobacterium used for the formulation of Medicago sativa bionoculants, and SmNirK plays a key role in this symbiosis through the denitrification pathway. The structure of SmNirK, solved at a resolution of 2.5Å, showed a striking resemblance with the overall structure of the well-known Class I NirKs composed of two Greek key β-barrel domains. The activity of SmNirK is ~12% of the activity reported for classical NirKs, which could be attributed to several factors such as subtle structural differences in the secondary proton channel, solvent accessibility of the substrate channel, and that the denitrifying activity has to be finely regulated within the endosymbiont. In vitro kinetics performed in homogenous and heterogeneous media showed that both SmPaz1 and SmPaz2, which are coded in different regions of the genome, donate electrons to SmNirK with similar performance. Even though the energetics of the interprotein electron transfer (ET) process is not favorable with either electron donors, adduct formation mediated by conserved residues allows minimizing the distance between the copper centers involved in the interprotein ET process.

How tree species with contrasting biological nitrification inhibition capacity influence denitrifier activity and abundance? Insights from reciprocal transfers of soil

How tree species with contrasting biological nitrification inhibition capacity influence denitrifier activity and abundance? Insights from reciprocal transfers of soil

Stimulation of N2 O emission via bacterial denitrification driven by acidification in estuarine sediments.

Ocean acidification in nitrogen-enriched estuaries has raised global concerns. For decades, biotic and abiotic denitrification in estuarine sediments has been regarded as the major ways to remove reactive nitrogen, but they occur at the expense of releasing greenhouse gas nitrous oxide (N2 O). However, how these pathways respond to acidification remains poorly understood. Here we performed a N2 O isotopocules analysis coupled with respiration inhibition and molecular approaches to investigate the impacts of acidification on bacterial, fungal, and chemo-denitrification, as well as N2 O emission, in estuarine sediments through a series of anoxic incubations. Results showed that acidification stimulated N2 O release from sediments, which was mainly mediated by the activity of bacterial denitrifiers, whereas in neutral environments, N2 O production was dominated by fungi. We also found that the contribution of chemo-denitrification to N2 O production cannot be ignored, but was not significantly affected by acidification. The mechanistic investigation further demonstrated that acidification changed the keystone taxa of sedimentary denitrifiers from N2 O-reducing to N2 O-producing ones and reduced microbial electron-transfer efficiency during denitrification. These findings provide novel insights into how acidification stimulates N2 O emission and modulates its pathways in estuarine sediments, and how it may contribute to the acceleration of global climate change in the Anthropocene.

Molecular Screening of Nitrate Reductase Enzyme in Native Halophilic Bacteria of Iran

With the increased usage of nitrate fertilizers, the removal of their stable ionic and water-soluble end products is a challenge for human health. Several physicochemical methods have been examined for nitrate removal of water, but biological treatments are mostly preferred due to a higher efficiency and lower cost. To remove nitrogen from water, we investigated the potential of nitrate-reducing halophilic and halotolerant bacteria. A total of 50 strains from different saline and hypersaline environments of Iran, including the Incheboron wetland, Aran-Bidgol salt-lake, and Urmia endorheic salt-lake, were screened for nitrate reductase production. Among investigated bacteria, 60% and 19% of strains obtained from Urmia lake, and Incheboron wetland produced nitrate reductases, respectively. The nitrate reductase coding genes narG, and napA were analyzed in all strains with confirmed nitrate-reducing capacity. The napA gene was successfully amplified from a gram-negative halophilic strain, and the narG gene was detected in ten halophilic strains. Among nitrate-reducing isolates with the narG gene expression, the Kocuria rosea strain R3A34 showed the highest nitrate reductase production level. This strain was selected to optimize for its denitrifying activity. Results showed that 32°C, pH 7.0, NaCl 8% (w/v), and mannitol (as a carbon source) provide the optimal environmental conditions for the efficient production of nitrate reductase by the Kocuria rosea strain R3A34. As these are compatible with wastewaters conditions, this bacterium can be a proper candidate for bioremediation of wastewaters from nitrate pollutants.

Soil phosphorus accumulation in mountainous alpine grassland contributes to positive climate change feedback via nitrifier and denitrifier community

Mountainous alpine ecosystems are sensitive to global change, where soil nutrient content would potentially vary under current climate change background, and thus possibly influence the activity of nitrifiers and denitrifiers, as well as N2O emissions. However, within mountainous alpine ecosystems, the potential variation of soil nutrients under current global change and the consequence to N2O emission from nitrification and denitrification are still unclarified, hampering a comprehensive understanding of the feedback mechanisms between the nitrogen cycle and climate change. In order to fill this knowledge gap, we selected alpine grasslands at three different elevations and investigated the distribution and environmental drivers of nitrifiers and denitrifiers. The results showed that the lowest elevation site tended to have higher total phosphorus (TP) accumulation within the topsoil. The abundance of functional groups, emission of CO2 and N2O, and the N2O/CO2 ratio showed a decreasing trend along elevation. TP was the greatest influence on denitrifier composition (nosZ/narG and nirS/nirK ratios) and considerably influenced nitrifier composition (AOA/AOB ratio), and was significantly correlated to the N2O/CO2 ratio. In microcosms of soils from the highest elevation site, TP addition decreased the ratios of nosZ/narG, nirS/nirK, and AOA/AOB, and increased N2O/CO2 ratio and N2O emission, thus contributing to positive climate change feedback. This study indicates the potential for change within the nitrifier and denitrifier communities under current climate change, and highlights the role TP plays in governing nitrification and denitrification in mountainous alpine ecosystems.

Response of denitrification rate and denitrifiers to human-disturbance intensity and physicochemical factors of overlying water in a subtropical stream, China

Human activities have greatly increased the availability of bioactive nitrogen and, consequently, have altered the nitrogen cycle in riverine systems. Here, we studied denitrification rates and denitrifiers in river sediment and their relationships with aquatic physicochemical variables along a gradient of human-disturbance intensity in a subtropical stream in China. We used high-throughput sequencing to characterize denitrifying assemblages (denitrification genes narG, nirK, nirS and nosZ as functional markers). We found that denitrification rates were positively correlated with water temperature, and the primary factors affecting denitrifier assemblage species richness were water temperature and NO3−. Denitrification rate was negatively associated with narG and nirS gene-coding denitrifier species richness and associated (both positively and negatively) with the relative abundances of many of the dominant denitrifiers with narG and nirS genes. The potential active denitrifiers with narG and nirS genes predominantly occurred in the most intensely disturbed area within the catchment (which also recorded the highest temperatures), but denitrification rates in the intensely and slightly disturbed areas were similar. Our research found relationships between denitrifier assemblage alpha diversity and relative abundances of dominant species, environmental factors, and denitrification rates. In this study, we did not find human-disturbance intensity to be a direct influence on denitrification, likely because denitrification is a complex process influenced by both the environment and the microbial assemblage.

Coupling Between the Responses of Plants, Soil, and Microorganisms Following Grazing Exclusion in an Overgrazed Grassland.

Grazing exclusion is an effective management practice to restore grassland ecosystem functioning. However, little is known about the role of soil microbial communities in regulating grassland ecosystem functioning during long-term ecosystem restorations. We evaluated the recovery of a degraded semiarid grassland ecosystem in northern China by investigating plant and soil characteristics and the role of soil microbial communities in ecosystem functioning after 22 years of grazing exclusion. Grazing exclusion significantly increased the alpha diversity and changed the community structure of bacteria, but did not significantly affect the alpha diversity or community structure of fungi. The higher abundance of copiotrophic Proteobacteria and Bacteroidetes with grazing exclusion was due to the higher carbon and nutrient concentrations in the soil, whereas the high abundance of Acidobacteria in overgrazed soils was likely an adaptation to the poor environmental conditions. Bacteria of the Sphingomonadaceae family were associated with C cycling under grazing exclusion. Bacteria of the Nitrospiraceae family, and especially of the Nitrospira genus, played an important role in changes to the N cycle under long-term exclusion of grazing. Quantitative PCR further revealed that grazing exclusion significantly increased the abundance of nitrogen fixing bacteria (nifH), ammonia oxidizers (AOA and AOB), and denitrifying bacteria (nirK and nosZ1). Denitrifying enzyme activity (DEA) was positively correlated with abundance of denitrifying bacteria. The increase in DEA under grazing exclusion suggests that the dependence of DEA on the availability of NO3– produced is due to the combined activity of ammonia oxidizers and denitrifiers. Our findings indicate that decades-long grazing exclusion can trigger changes in the soil bacterial diversity and composition, thus modulating the restoration of grassland ecosystem functions, carbon sequestration and soil fertility.

Slow pyrolysis pine wood-derived biochar reduces nitrous oxide production from surface but not subsurface soil

Soil amended with biochar is expected to produce less nitrous oxide (N2O), although this may depend on nitrate (NO3-N) availability. Our objective was to determine how pine wood biochar, slow pyrolyzed at 500 °C, affects N2O production in soil having different denitrification potentials with variable NO3-N concentrations under controlled laboratory conditions. Sandy loam surface soils (0–30 cm, pH 5.7) and sandy clay loam subsurface soils (40–60 cm, pH 5.6) were amended with four biochar rates (0, 10, 20, and 30 g·kg−1), two nitrogen fertilizer rates (0 and 100 mg·kg−1 NO3-N) and two acetylene levels (0% and 10% headspace), arranged as a full factorial. Soil moisture content was adjusted to 80% water-filled pore space, and flasks were incubated at 20 °C for 30 h. Headspace gas was collected from each flask at 25, 26, 28, and 30 h. There was a significant reduction in N2O production with an increasing rate of biochar in the surface soil but not in the subsurface soil. On average, less N2O was produced in the subsurface soil than in the surface soil. As the NO3-N concentration was not a limiting factor for denitrification, the most likely explanation was that denitrifier activity was influenced by the availability of soluble organic carbon in the soil–biochar mixtures. We recommend further study of the coupled carbon–nitrogen transformations during denitrification to understand how biochar influences soil N2O production in sandy loam soils.

Differential responses of soil nitrogen‐oxide emissions to organic substitution for synthetic fertilizer and biochar amendment in a subtropical tea plantation

AbstractTropical and subtropical acidic soils have been well documented as hotspots of global soil nitrogen (N) oxide (i.e., nitrous oxide (N2O) and nitric oxide (NO) emissions). While the effectiveness of possible mitigation options has been extensively examined in croplands, little is known about their effectiveness in reducing N‐oxide emissions from acidic soils of rapidly expanding tea plantations in China. Here, we conducted a 2‐year field experiment to investigate how organic substitution for synthetic fertilizer and biochar amendment affect soil N‐oxide emissions from a subtropical tea plantation. Across the 2‐year measurement period, full organic substitution for synthetic fertilizer significantly increased N2O emissions by an average of 17% while had a lower NO emission compared to synthetic fertilizer alone. Our global meta‐analysis further revealed that full or partial organic fertilizer substitution resulted in a 29% (95% confidence interval: 5%–60%) increase of N2O emissions from acidic soils. In contrast, irrespective of fertilizer type, biochar amendment significantly reduced N2O emissions by 14% in the first but not second experimental year, suggesting a transient effect. The trade‐off effect of full organic substitution on N2O and NO emissions may be attributed to the favorable conditions for N2O production due to the stimulated activity of nitrifiers and denitrifiers. The suppression of N2O emission following biochar amendment was probably due to promoted further reduction of N2O to dinitrogen. The fertilizer‐induced emission factor (EF) of N2O (2.1%) in the tea plantation was greater than the current IPCC default value, but the EF of NO (0.8%) was comparable to the global estimate. Taken together, while biochar amendment could have mitigation potential, cautions are needed when applying organic substitution for synthetic fertilizer as mitigation options for acidic soils as hotspots of N‐oxide emissions.

Diversity of active anaerobic ammonium oxidation (ANAMMOX) and nirK-type denitrifying bacteria in macrophyte roots in a eutrophic wetland

Wetland eutrophication is a global ecological problem. Anaerobic oxidation of ammonium (ANAMMOX) and denitrification are the two major pathways of nitrogen removal in wetlands. However, little is known about the associated bacteria that are present around emergent macrophytes and their transcriptional activities. Here, we examine the transcriptional activity of bacteria that perform ANAMMOX and denitrification in the roots and vegetated sediments of three emergent macrophytes and in unvegetated sediment from a eutrophic wetland. Roots and vegetated sediments of three emergent macrophytes (Phragmites australis, Typha angustifolia, and Scirpus triqueter) and unvegetated sediment were collected from Wuliangsuhai wetland, China. The community structure and transcriptional activity of ANAMMOX and denitrifying bacteria were examined at the DNA and RNA levels. Additionally, the sediment physicochemical properties were determined. ANAMMOX (hzsB) and denitrifying bacteria (nirK, nirS, and nirSII) were detected in all sediments and roots at the DNA level. However, at the RNA level, root-associated ANAMMOX bacteria, including Candidatus Jettenia and Candidatus Brocadia, were detected only in the roots of P. australis. In contrast, nirK-carrying active denitrifiers, such as uncultured bacteria, Sinorhizobium, Rhizobiales, Mesorhizobium, Rhizobium, Devosia, and Bosea (all Rhizobiales), were detected in the roots of T. angustifolia and S. triqueter. The reduction of nitrogen concentration in the sediments of the vegetation zone was probably associated with active ANAMMOX bacteria and denitrifiers. Active root-associated ANAMMOX bacteria and nirK-type denitrifiers were influenced by plant species, and they likely contribute to nitrogen removal in eutrophic wetlands.