Nitrogen-cycling processes under long-term compound heavy metal(loids) pressure around a gold mine: Stimulation of nitrite reduction.

Mining and tailings deposition can cause serious heavy metal(loids) pollution to the surrounding soil environment. Soil microorganisms adapt their metabolism to such conditions, driving alterations in soil function. This study aims to elucidate the response patterns of nitrogen-cycling microorganisms under long-term heavy metal(loids) exposure. The results showed that the diversity and abundance of nitrogen-cycling microorganisms showed negative feedback to heavy metal(loids) concentrations. Denitrifying microorganisms were shown to be the dominant microorganisms with over 60% of relative abundance and a complex community structure including 27 phyla. Further, the key bacterial species in the denitrification process were calculated using a random forest model, where the top three key species (Pseudomonas stutzei, Sphingobium japonicum and Leifsonia rubra) were found to play a prominent role in nitrite reduction. Functional gene analysis and qPCR revealed that nirK, which is involved in nitrite reduction, significantly accumulated in the most metal-rich soil with the increase of absolute abundance of 63.86%. The experimental results confirmed that the activity of nitrite reductase (Nir) encoded by nirK in the soil was increased at high concentrations of heavy metal(loids). Partial least squares-path model identified three potential modes of nitrite reduction processes being stimulated by heavy metal(loids), the most prominent of which contributed to enhanced nirK abundance and soil Nir activity through positive stimulation of key species. The results provide new insights and preliminary evidence on the stimulation of nitrite reduction processes by heavy metal(loids).

Nitrifying niche in estuaries is expanded by the plastisphere.

The estuarine plastisphere, a novel ecological habitat in the Anthropocene, has garnered global concerns. Recent geochemical evidence has pointed out its potential role in influencing nitrogen biogeochemistry. However, the biogeochemical significance of the plastisphere and its mechanisms regulating nitrogen cycling remain elusive. Using 15N- and 13C-labelling coupled with metagenomics and metatranscriptomics, here we unveil that the plastisphere likely acts as an underappreciated nitrifying niche in estuarine ecosystems, exhibiting a 0.9 ~ 12-fold higher activity of bacteria-mediated nitrification compared to surrounding seawater and other biofilms (stone, wood and glass biofilms). The shift of active nitrifiers from O2-sensitive nitrifiers in the seawater to nitrifiers with versatile metabolisms in the plastisphere, combined with the potential interspecific cooperation of nitrifying substrate exchange observed among the plastisphere nitrifiers, collectively results in the unique nitrifying niche. Our findings highlight the plastisphere as an emerging nitrifying niche in estuarine environment, and deepen the mechanistic understanding of its contribution to marine biogeochemistry.

[Vertical Distribution Characteristics and Driving Factors of Bacterioplankton and Nitrogen Phosphorus Cycle Genes in Danjiangkou Reservoir].

Danjiangkou Reservoir is a critical water source for the South-to-North Water Diversion Project, which harbors a diverse bacterioplankton community with varying depths, and the understanding of its nitrogen and phosphorus cycle and associated driving factors remains limited. In this study, we selected five ecological sites within Danjiangkou Reservoir and conducted metagenomics analysis to investigate the vertical distribution of bacterioplankton communities in the surface, middle, and bottom layers. Furthermore, we analyzed and predicted the function of nitrogen and phosphorus cycles, along with their driving factors. Our findings revealed the dominance of Proteobacteria, Actinobacteria, and Planctomycetes in the Danjiangkou Reservoir. Significant differences were observed in the structure of bacterioplankton communities across different depths, with temperature （T）, oxidation-reduction potential （ORP）, dissolved oxygen （DO）, and Chla identified as primary factors influencing the bacterioplankton composition. Analysis of nitrogen cycle functional genes identified 39 genes, including gltB, glnA, gltD, gdhA, NRT, etc., which were involved in seven main pathways, encompassing nitrogen fixation, nitrification, denitrification, and dissimilatory nitrate reduction. Phosphorus cycle function gene analysis identified 54 genes, including pstS, ppx-gppA, glpQ, ppk1, etc., primarily participating in six main pathways, including organic P mineralization, inorganic P solubilization, and regulatory. Cluster analysis indicated that different depths were significant factors influencing the composition and abundance of nitrogen and phosphorus cycle functional genes. The composition and abundance of nitrogen and phosphorus cycle functional genes in the surface and bottom layers differed and were generally higher than those in the middle layer. Deinococcus, Hydrogenophaga, Limnohabitans, Clavibacter, and others were identified as key species involved in the nitrogen and phosphorus cycle. Additionally, we found significant correlations between nitrogen and phosphorus cycle functional genes and environmental factors such as DO, pH, T, total dissolved solids （TDS）, electrical conductivity （EC）, and Chla. Furthermore, the content of these environmental factors exhibited depth-related changes in the Danjiangkou Reservoir, resulting in a distinct vertical distribution pattern of bacterioplankton nitrogen and phosphorus cycle functional genes. Overall, this study sheds light on the composition, function, and influencing factors of bacterioplankton communities across different layers of Danjiangkou Reservoir, offering valuable insights for the ecological function and diversity protection of bacterioplankton in this crucial reservoir ecosystem.

Effect of nitrogen addition on soil net nitrogen mineralization in topsoil and subsoil regulated by soil microbial properties and mineral protection: Evidence from a long-term grassland experiment.

Soil net nitrogen mineralization (Nmin), a microbial-mediated conversion of organic to inorganic N, is critical for grassland productivity and biogeochemical cycling. Enhanced atmospheric N deposition has been shown to substantially increase both plant and soil N content, leading to a major change in Nmin. However, the mechanisms underlying microbial properties, particularly microbial functional genes, which drive the response of Nmin to elevated N deposition are still being discussed. Besides, it is still uncertain whether the relative importance of plant carbon (C) input, microbial properties, and mineral protection in regulating Nmin under continuous N addition would vary with the soil depth. Here, based on a 13-year multi-level field N addition experiment conducted in a typical grassland on the Loess Plateau, we elucidated how N-induced changes in plant C input, soil physicochemical properties, mineral properties, soil microbial community, and the soil Nmin rate (Rmin)-related functional genes drove the responses of Rmin to N addition in the topsoil and subsoil. The results showed that Rmin increased significantly in both topsoil and subsoil with increasing rates of N addition. Such a response was mainly dominated by the rate of soil nitrification. Structural equation modeling (SEM) revealed that a combination of microbial properties (functional genes and diversity) and mineral properties regulated the response of Rmin to N addition at both soil depths, thus leading to changes in the soil N availability. More importantly, the regulatory impacts of microbial and mineral properties on Rmin were depth-dependent: the influences of microbial properties weakened with soil depth, whereas the effects of mineral protection enhanced with soil depth. Collectively, these results highlight the need to incorporate the effects of differential microbial and mineral properties on Rmin at different soil depths into the Earth system models to better predict soil N cycling under further scenarios of N deposition.

[Spatial Patterns of Soil Bacterial Communities and N-cycling Functional Groups Along an Altitude Gradient in Datong River Basin].

The altitude distribution patterns of soil microorganisms and their driving mechanisms are crucial for understanding the consequences of climate change on terrestrial ecosystems. There is an obvious altitude difference in Datong River Basin in the Qilian Mountains. Two spatial scale transections were set up along the mountain slope (with altitude spanning 1 000 m) and the mainstream direction (with altitude spanning 300-500 m), respectively. The distribution characteristics of the soil bacterial community structure and diversity along the altitude gradients were examined using high-throughput sequencing technology. Based on the FAPROTAX database, the altitude distribution patterns of nitrogen cycling functional groups were analyzed to investigate the major environmental factors influencing the altitude distribution patterns of soil bacterial communities. The findings revealed that:① Soil physicochemical characteristics varied significantly with altitude. The content of total nitrogen (TN) and nitrate nitrogen (NO3-) were positively correlated with the altitude (P < 0.01), whereas the soil bulk density and pH were negatively connected (P < 0.001). ② The abundance of OTU increased significantly along the altitude (P < 0.01), and the richness and diversity indices increased along the altitude, although the trend was not statistically significant (P > 0.05). ③ The predominant bacterial communities were Acidobacteria, Proteobacteria, and Bacteroidetes, and as altitude climbed, their relative abundances varied between increasing, decreasing, and slightly decreasing, respectively. ④ The nitrogen cycling processes involved 13 functional groups, primarily nitrification, aerobic ammonia oxidation, aerobic nitrite oxidation, etc. As the altitude increased, the response law changed, with an increase in the abundance of nitrobacteria (P < 0.01), a slight increase in the abundance of aerobic ammonia-oxidizing bacteria and nitrite-oxidizing bacteria, and a hump-back tendency in bacteria abundance for nitrogen respiration. ⑤ Redundancy analysis revealed that the key determinants influencing soil bacterial populations at the phylum level were altitude, pH, and the content of NH4+. Mantel analysis showed that the dominant groups of soil bacterial nitrogen cycling were all statistically and significantly driven by altitude (P < 0.01). ⑥ The α-diversity of the bacterial community with increasing altitude were both increased along the mountain slope and the mainstream direction, but the soil properties, the abundance of N-cycling functional groups, and the main environmental factors differed. Therefore, it is of great significance to explore the altitude distribution pattern of soil microorganisms at different spatial scales.

Aridity threshold for alpine soil nitrogen isotope signature and ecosystem nitrogen cycling.

Determination of tipping points in nitrogen (N) isotope (δ15N) natural abundance, especially soil δ15N, with increasing aridity, is critical for estimating N-cycling dynamics and N limitation in terrestrial ecosystems. However, whether there are linear or nonlinear responses of soil δ15N to increases in aridity and if these responses correspond well with soil N cycling remains largely unknown. In this study, we investigated soil δ15N and soil N-cycling characteristics in both topsoil and subsoil layers along a drought gradient across a 3000-km transect of drylands on the Qinghai-Tibetan Plateau. We found that the effect of increasing aridity on soil δ15N values shifted from negative to positive with thresholds at aridity index (AI) = 0.27 and 0.29 for the topsoil and subsoil, respectively, although soil N pools and N transformation rates linearly decreased with increasing aridity in both soil layers. Furthermore, we identified markedly different correlations between soil δ15N and soil N-cycling traits above and below the AI thresholds (0.27 and 0.29 for topsoil and subsoil, respectively). Specifically, in wetter regions, soil δ15N positively correlated with most soil N-cycling traits, suggesting that high soil δ15N may result from the "openness" of soil N cycling. Conversely, in drier regions, soil δ15N showed insignificant relationships with soil N-cycling traits and correlated well with factors, such as soil-available phosphorus and foliage δ15N, demonstrating that pathways other than typical soil N cycling may dominate soil δ15N under drier conditions. Overall, these results highlight that different ecosystem N-cycling processes may drive soil δ15N along the aridity gradient, broadening our understanding of N cycling as indicated by soil δ15N under changing drought regimes. The aridity threshold of soil δ15N should be considered in terrestrial N-cycling models when incorporating 15N isotope signals to predict N cycling and availability under climatic dryness.

Biochar immobilized hydrolase degrades PET microplastics and alleviates the disturbance of soil microbial function via modulating nitrogen and phosphorus cycles.

Microplastics (MPs) pose an emerging threat to soil ecological function, yet effective solutions remain limited. This study introduces a novel approach using magnetic biochar immobilized PET hydrolase (MB-LCC-FDS) to degrade soil polyethylene terephthalate microplastics (PET-MPs). MB-LCC-FDS exhibited a 1.68-fold increase in relative activity in aquatic solutions and maintained 58.5 % residual activity after five consecutive cycles. Soil microcosm experiment amended with MB-LCC-FDS observed a 29.6 % weight loss of PET-MPs, converting PET into mono(2-hydroxyethyl) terephthalate (MHET). The generated MHET can subsequently be metabolized by soil microbiota to release terephthalic acid. The introduction of MB-LCC-FDS shifted the functional composition of soil microbiota, increasing the relative abundances of Microbacteriaceae and Skermanella while reducing Arthobacter and Vicinamibacteraceae. Metagenomic analysis revealed that MB-LCC-FDS enhanced nitrogen fixation, P-uptake and transport, and organic-P mineralization in PET-MPs contaminated soil, while weakening the denitrification and nitrification. Structural equation model indicated that changes in soil total carbon and Simpson index, induced by MB-LCC-FDS, were the driving factors for soil carbon and nitrogen transformation. Overall, this study highlights the synergistic role of magnetic biochar-immobilized PET hydrolase and soil microbiota in degrading soil PET-MPs, and enhances our understanding of the microbiome and functional gene responses to PET-MPs and MB-LCC-FDS in soil systems.

Extending grazing time during the warm season can reduce P limitation and increase the N cycling rate in arid desert steppes.

Ecological stoichiometry serves as a valuable tool for comprehending biogeochemical cycles within grassland ecosystems. The impact of grazing time on the concentration and stoichiometric characteristics of carbon (C), nitrogen (N), and phosphorus (P) in desert steppe ecosystems remains ambiguous. This research was carried out in a desert grassland utilizing a completely randomized experimental design. Four distinct grazing time treatments were implemented: fenced grassland (FG, control), delay to start and early to end grazing grassland (DEG), delay to start grazing grassland (DG), and traditional grazing grassland (TG). The patterns of C, N, and P concentrations and their stoichiometry in various components of the ecosystem, as well as their driving factors under different grazing times were examined. The results showed that grazing time positively influenced C and N concentrations in leaves, while negatively affecting N concentrations in roots. TG had a significant positive effect on soil P concentrations but a negative effect on soil C:P and N:P ratios. Plant C:N, C:P, and N: P ratios were mainly influenced by N and P. The soil C:N ratio was primarily influenced by soil N, the soil C:P ratio was affected by both soil C and P, and the soil N:P ratio was influenced by both soil N and P. The growth of plants in desert steppes is mainly limited by P; however, as grazing time increased, P limitation gradually decreased and the N cycling rate increased. C-N, C-P, and N-P in various plant organs and soils demonstrated significant anisotropic growth relationships at different grazing times. Soil organic carbon, pH, and soil total phosphorus were the main driving factors that affected changes in ecological C:N:P stoichiometry. These results will help improve grassland management and anticipate the response of grassland systems to external disturbances with greater accuracy.

Differential impacts of polyethylene microplastic and additives on soil nitrogen cycling: A deeper dive into microbial interactions and transformation mechanisms.

The impact of microplastics and their additives on soil nutrient cycling, particularly through microbial mechanisms, remains underexplored. This study investigated the effects of polyethylene microplastics, polyethylene resin, and plastic additives on soil nitrogen content, physicochemical properties, nitrogen cycling functional genes, microbial composition, and nitrogen transformation rates. Results showed that all amendments increased total nitrogen but decreased dissolved total nitrogen. Polyethylene microplastics and additives increased dissolved organic nitrogen, while polyethylene resin reduced it and exhibited higher microbial biomass. Amendments reduced or did not change inorganic nitrogen levels, with additives showing the lowest values. Polyethylene resin favored microbial nitrogen immobilization, while additives were more inhibitory. Amendment type and content significantly interacted with nitrogen cycling genes and microbial composition. Distinct functional microbial biomarkers and network structures were identified for different amendments. Polyethylene microplastics had higher gross ammonification, nitrification, and immobilization rates, followed by polyethylene resin and additives. Nitrogen transformation was driven by multiple functional genes, with Proteobacteria playing a significant role. Soil physicochemical properties affected nitrogen content through transformation rates, with C/N ratio having an indirect effect and water holding capacity directly impacting it. In summary, plastic additives, compared to polyethylene microplastics and resin, are less conducive to nitrogen degradation and microbial immobilization, exert significant effects on microbial community structure, inhibit transformation rates, and ultimately impact nitrogen cycling.

Organic fertilizer application promotes the soil nitrogen cycle and plant starch and sucrose metabolism to improve the yield of Pinellia ternata.

Pinellia ternata (Thunb.) Breit is a traditional Chinese medicine with important pharmacological effects. However, its cultivation is challenged by soil degradation following excessive use of chemical fertilizer. We conducted an experiment exploring the effects of replacing chemical fertilizers with organic fertilizers (OF) on the growth and yield of P. ternata, as well as on the soil physicochemical properties and microbial community composition using containerized plants. Six fertilization treatments were evaluated, including control (CK), chemical fertilizer (CF), different proportions of replacing chemical fertilizer with organic fertilizer (OM1-4). Containerized P. ternata plants in each OF treatment had greater growth and yield than the CK and CF treatments while maintaining alkaloid content. The OM3 treatment had the greatest yield among all treatments, with an increase of 42.35% and 44.93% compared to the CK and CF treatments, respectively. OF treatments improved soil quality and fertility by enhancing the activities of soil urease (S-UE) and sucrase (S-SC) enzymes while increasing soil organic matter and trace mineral elements. OF treatments increased bacterial abundance and changed soil community structure. In comparison to the CK microbial groups enriched in OM3 were OLB13, Vicinamibacteraceae, and Blrii41. There were also changes in the abundance of gene transcripts among treatments. The abundance of genes involved in the nitrogen cycle in the OM3 has increased, specifically promoting the transformation of N-NO3- into N-NH4+, a type of nitrogen more easily absorbed by P. ternata. Also, genes involved in "starch and sucrose metabolism" and "plant hormone signal transduction" pathways were positively correlated to P. ternata yield and were upregulated in the OM3 treatment. Overall, OF in P. ternata cultivation is a feasible practice in advancing sustainable agriculture and is potentially profitable in commercial production.

Hypoxia exerts greater impacts on shallow groundwater nitrogen cycling than seawater mixture in coastal zone.

There is no doubt that hypoxia and seawater mixture are profoundly affecting the global nitrogen (N) cycle. However, their mechanisms for altering N cycling patterns in shallow coastal groundwater are largely unknown. Here, we examined shallow groundwater N transformation characteristics (dissolved inorganic N and related chemical properties) in the coastal area of east and west Shenzhen City. Results showed that common hypoxic conditions exist in this study area. Ions/Cl- ratios indicated varying levels of saltwater mixture and sulfide formation across this study area. Dissolved oxygen (DO) affects the N cycle process by controlling the conditions of nitrification and the formation of sulfides. Salinity affects nitrification and denitrification processes by physiological effects, while sulfide impacts nitrification, denitrification, and dissimilatory nitrate reduction to ammonium (DNRA) processes through its own toxicity mechanism and the provision of electron donors for DNRA organisms. Redundancy analysis (RDA) results indicate that the influence magnitude is in the following order: DO > sulfide > salinity. Seawater mixture weakened the nitrification and denitrification of groundwater by changing salinity, while hypoxia and its controlled sulfide formation not only weaken nitrification and denitrification but also stimulated the DNRA process and promotes N regeneration. In this study area, hypoxia is considered to exert greater impacts on N cycling in the coastal shallow groundwater than seawater mixture. These findings greatly improve our understanding of the consequences of hypoxia and seawater mixture on coastal groundwater N cycling.

Removal efficiency and abundance of nitrogen cycling microorganism in three bio-matrix materials to treat swine wastewater.

A bio-matrix material (BMM) system is used to pretreat swine wastewater and reduce the nitrogen (N) concentration to the tolerance range of plants in constructed wetlands. In this study, rice straw (RS), wheat straw (WS), and corn stalk (CS) were applied to treat pollutants from swine wastewater, respectively. This one year-long field experiment make up for the lack of long-term experiments and mechanistic investigations of BMM. The pollutant removal efficiency, degradation process of crop straw, and the abundance of nitrogen cycling genes were determined in different BMM systems. The results showed that the removal efficiency of COD, TN, NH4+, and NO3- was the best in the initial 6 months. Furthermore, RS and WS exhibited favorable annual removal efficiency of TN and NH4+, which were 32.81% and 32.99%, 35.3% and 34.97%, respectively. Moreover, the removal efficiency of COD was 30.81% in three BMM systems. Meanwhile, it was found that the dry matter (DM) degradation of crop straws was fast in the first 4-5 months. The degradation rates of cellulose, hemicellulose, and lignin were 94.19%, 94.36%, and 87.32%, respectively, in 1 year. The abundance of nitrogen cycling genes significantly increased by adding BMM, compared with CK (P < 0.05). This showed the abundance of the hzsB gene in RS was the highest, while nirK, nirS, AOA, and AOB were the highest in WS. The addition of RS and WS was better than that of CS in promoting the abundance of nitrogen cycling microorganisms. The results indicated that adding BMM could enhance the anaerobic ammonia oxidation, nitrification, and denitrification. This study not only extends our comprehension of BMM mechanisms in swine wastewater treatment but also serves as a guiding light for numerous farms in similar climate regions.

Distinct microbial nitrogen cycling processes in the deepest part of the ocean.

The Mariana Trench (MT) is the deepest part of the ocean on Earth. Previous studies have described the microbial community structures and functional potential in the seawater and surface sediment of MT. Still, the metabolic features and adaptation strategies of the microorganisms involved in nitrogen cycling processes are poorly understood. In this study, comparative metagenomic approaches were used to study microbial nitrogen cycling in three MT habitats, including hadal seawater [9,600-10,500 m below sea level (mbsl)], surface sediments [0-46 cm below seafloor (cmbsf) at a water depth between 7,143 and 8,638 mbsl], and deep sediments (200-306 cmbsf at a water depth of 8,300 mbsl). We identified five new nitrite-oxidizing bacteria (NOB) lineages that had adapted to the oligotrophic MT slope sediment, via their CO2 fixation capability through the reductive tricarboxylic acid (rTCA) or Calvin-Benson-Bassham (CBB) cycle; an anammox bacterium might perform aerobic respiration and utilize sedimentary carbohydrates for energy generation because it contains genes encoding type A cytochrome c oxidase and complete glycolysis pathway. In seawater, abundant alkane-oxidizing Ketobacter species can fix inert N2 released from other denitrifying and/or anammox bacteria. This study further expands our understanding of microbial life in the largely unexplored deepest part of the ocean. IMPORTANCE: The metabolic features and adaptation strategies of the nitrogen cycling microorganisms in the deepest part of the ocean are largely unknown. This study revealed that anammox bacteria might perform aerobic respiration in response to nutrient limitation or O2 fluctuations in the Mariana Trench sediments. Meanwhile, an abundant alkane-oxidizing Ketobacter species could fix N2 in hadal seawater. This study provides new insights into the roles of hadal microorganisms in global nitrogen biogeochemical cycles. It substantially expands our understanding of the microbial life in the largely unexplored deepest part of the ocean.

Effects of climate warming on soil nitrogen cycles and bamboo growth in core giant panda habitat.

Climate change can pose a significant threat to terrestrial ecosystems by disrupting the circulation of soil nitrogen. However, experimental analyses on the effect of climate change on soil nitrogen cycles and the implications for the conservation of key wildlife species (i.e., the giant panda, Ailuropoda melanoleuca) remain understudied. We investigated the effects of a 1.5 °C, 3 °C, and 4.5 °C temperature increase on nitrogen distribution in different soil layers of bamboo forest via an in-situ experiment and assessed the implications for the growth and survival of arrow bamboo (Bashania faberi), a critical food resource for giant pandas. Our results showed that warming treatments generally increased soil N content, while effects differed between surface soil and subsurface soil and at different warming treatments. Particularly an increase of 1.5 °C raised the subsurface soil NO3-N content, as well as the content of N in bamboo leaves. We found a significant positive correlation between the subsurface soil NO3-N content and the N content of arrow bamboo. An increase of 3-4.5 °C raised the content of total N and NO3-N in the surface soil and led to a reduction in the total aboveground biomass and survival rate of arrow bamboo. Limited warming (e.g., the increase of 0-1.5 °C) may promote the soil N cycle, raise the N-acetylglucosaminidase (NAG) enzyme activity, increase NO3-N in subsurface soil, increase the N content of bamboo, and boost the biomass of bamboo - all of which could be beneficial to giant panda survival. However, higher warming (e.g., an increase of 3-4.5 °C) resulted in mass death of bamboo and a large reduction in aboveground biomass. Our findings provide a cautiously optimistic scenario for bamboo forest ecosystems under low levels of warming over a short period of time, but risks from higher levels of warming may be serious, especially considering the unpredictability of global climatic change.

Nitrate reduction to ammonium: a phylogenetic, physiological, and genetic aspects in Prokaryotes and eukaryotes.

The microbe-mediated conversion of nitrate (NO3-) to ammonium (NH4+) in the nitrogen cycle has strong implications for soil health and crop productivity. The role of prokaryotes, eukaryotes and their phylogeny, physiology, and genetic regulations are essential for understanding the ecological significance of this empirical process. Several prokaryotes (bacteria and archaea), and a few eukaryotes (fungi and algae) are reported as NO3- reducers under certain conditions. This process involves enzymatic reactions which has been catalysed by nitrate reductases, nitrite reductases, and NH4+-assimilating enzymes. Earlier reports emphasised that single-cell prokaryotic or eukaryotic organisms are responsible for this process, which portrayed a prominent gap. Therefore, this study revisits the similarities and uniqueness of mechanism behind NO3- -reduction to NH4+ in both prokaryotes and eukaryotes. Moreover, phylogenetic, physiological, and genetic regulation also shed light on the evolutionary connections between two systems which could help us to better explain the NO3--reduction mechanisms over time. Reports also revealed that certain transcription factors like NtrC/NtrB and Nit2 have shown a major role in coordinating the expression of NO3- assimilation genes in response to NO3- availability. Overall, this review provides a comprehensive information about the complex fermentative and respiratory dissimilatory nitrate reduction to ammonium (DNRA) processes. Uncovering the complexity of this process across various organisms may further give insight into sustainable nitrogen management practices and might contribute to addressing global environmental challenges.

Organic fertilization reduces nitrous oxide emission by altering nitrogen cycling microbial guilds favouring complete denitrification at soil aggregate scale.

Agricultural management practices can induce changes in soil aggregation structure that alter the microbial nitrous oxide (N2O) production and reduction processes occurring at the microscale, leading to large-scale consequences for N2O emissions. However, the mechanistic understanding of how organic fertilization affects these context-dependent small-scale N2O emissions and associated key nitrogen (N) cycling microbial communities is lacking. Here, denitrification gas (N2O, N2) and potential denitrification capacity N2O/(N2O + N2) were assessed by automated gas chromatography in different soil aggregates (>2 mm, 2-0.25 and <0.25 mm), while associated microbial communities were assessed by sequencing and qPCR of N2O-producing (nirK and nirS) and reducing (nosZ clade I and II) genes. The results indicated that organic fertilization reduced N2O emissions by enhancing the conversion of N2O to N2 in all aggregate sizes. Moreover, potential N2O production and reduction hotspots occurred in smaller soil aggregates, with the degree depending on organic fertilizer type and application rate. Further, significantly higher abundance and diversity of nosZ clades relative to nirK and nirS revealed complete denitrification promoted through selection of denitrifying communities at microscales favouring N2O reduction. Communities associated with high and low emission treatments form modules with specific sequence types which may be diagnostic of emission levels. Taken together, these findings suggest that organic fertilizers reduced N2O emissions through influencing soil factors and patterns of niche partitioning between N2O-producing and reducing communities within soil aggregates, and selection for communities that overall are more likely to consume than emit N2O. These findings are helpful in strengthening the ability to predict N2O emissions from agricultural soils under organic fertilization as well as contributing to the development of net-zero carbon strategies for sustainable agriculture.

Analysis of effects and factors linked to soil microbial populations and nitrogen cycling under long-term biosolids application.

Information about impacts of long-term biosolids application on soil microbial populations and functional groups and N cycling is important for evaluating soil health and agroecosystem sustainability under long-term biosolids application. Mine spoil plots received annual biosolids application from 1973 to 2010 at low (16.8 Mg ha-1 yr-1), medium (33.6 Mg ha-1 yr-1), and high rates (67.2 Mg ha-1 yr-1). A no-biosolids control received chemical fertilizer at the agronomic rate. Soil samples were collected in three seasons per year spanning 2003-2005 for measuring soil moisture, pH, soil organic C (SOC), total and extractable heavy metals (Cd, Cu, Ni, Zn), NO3-, N mineralization potential (NMP), microbial biomass C (MBC), and populations of three N-cycling bacteria (NCB) groups: ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), and denitrifying bacteria (DNB). Soil samples were collected again in 2008 and 2010 for quantifying total and extractable heavy metals, and in 2018 (eight years after biosolids applications ended) for measuring SOC, MBC, NMP, and microbial respiration. During 2003-2005, mean MBC was 315, 554, 794, and 1001 mg kg-1 in the control, low, medium, and high biosolids treatments, respectively. Populations of NCB did not differ among treatments. Biosolids application increased total and extractable metal concentrations but the effect of biosolids rates were much lower on extractable than total concentrations. Soil extractable Cd and Cu concentrations decreased from medium to high applications, likely due to complexing with biosolids organic matter. Partial least squares regression analysis identified a strong positive effect on MBC of SOC and a weak negative effect of Cu, explaining the strong net positive effect of biosolids on MBC. In 2018, the medium and high biosolids treatments maintained higher SOC, MBC, NMP, and microbial respiration than the control. This study provided further evidence that long-term biosolids application has positive effects on soil microbes that persist for years after ending application.

Biochar and peat amendments affect nitrogen retention, microbial capacity and nitrogen cycling microbial communities in a metal and polycyclic aromatic hydrocarbon contaminated urban soil.

Soil contaminants may restrict soil functions. A promising soil remediation method is amendment with biochar, which has the potential to both adsorb contaminants and improve soil health. However, effects of biochar amendment on soil-plant nitrogen (N) dynamics and N cycling microbial guilds in contaminated soils are still poorly understood. Here, a metal- and polycyclic aromatic hydrocarbon (PAH) contaminated soil was amended with either biochar (0, 3, 6 % w/w) and/or peat (0, 1.5, 3 % w/w) in a full-factorial design and sown with perennial ryegrass in an outdoor field trial. After three months, N and the stable isotopic ratio δ15N was measured in soil, roots and leaves, along with microbial responses. Aboveground grass biomass decreased by 30 % and leaf N content by 20 % with biochar, while peat alone had no effect. Peat in particular, but also biochar, stimulated the abundance of microorganisms (measured as 16S rRNA gene copy number) and basal respiration. Microbial substrate utilization (MicroResp™) was altered differentially, as peat increased respiration of all carbon sources, while for biochar, respiration of carboxylic acids increased, sugars decreased, and was unaffected for amino acids. Biochar increased the abundance of ammonia oxidizing archaea, while peat stimulated ammonia oxidizing bacteria, Nitrobacter-type nitrite oxidizers and comB-type complete ammonia oxidizers. Biochar and peat also increased nitrous oxide reducing communities (nosZI and nosZII), while peat alone or combined with biochar also increased abundance of nirK-type denitrifiers. However, biochar and peat lowered leaf δ15N by 2-4 ‰, indicating that processes causing gaseous N losses, like denitrification and ammonia volatilization, were reduced compared to the untreated contaminated soil, probably an effect of biotic N immobilization. Overall, this study shows that in addition to contaminant stabilization, amendment with biochar and peat can increase N retention while improving microbial capacity to perform important soil functions.

Does long-term drought or repeated defoliation affect seasonal leaf N cycling in young beech trees?

Forest trees adopt effective strategies to optimize nitrogen (N) use through internal N recycling. In the context of more recurrent environmental stresses due to climate change, the question remains of whether increased frequency of drought or defoliation threatens this internal N recycling strategy. We submitted 8-year-old beech trees to 2 years of either severe drought (Dro) or manual defoliation (Def) to create a state of N starvation. At the end of the second year before leaf senescence, we labeled the foliage of the Dro and Def trees, as well as that of control (Co) trees, with 15N-urea. Leaf N resorption, winter tree N storage (total N, 15N, amino acids, soluble proteins) and N remobilization in spring were evaluated for the three treatments. Defoliation and drought did not significantly impact foliar N resorption or N concentrations in organs in winter. Total N amounts in Def tree remained close to those in Co tree, but winter N was stored more in the branches than in the trunk and roots. Total N amount in Dro trees was drastically reduced (-55%), especially at the trunk level, but soluble protein concentrations increased in the trunk and fine roots compared with Co trees. During spring, 15N was mobilized from the trunk, branches and twigs of both Co and Def trees to support leaf growth. It was only provided through twig 15N remobilization in the Dro trees, thus resulting in extremely reduced Dro leaf N amounts. Our results suggest that stress-induced changes occur in N metabolism but with varying severity depending on the constraints: within-tree 15N transport and storage strategy changed in response to defoliation, whereas a soil water deficit induced a drastic reduction of the N amounts in all the tree organs. Consequently, N dysfunction could be involved in drought-induced beech tree mortality under the future climate.