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Biological nitrogen (N) fixation, an important pathway of N, inputs from the atmosphere to Earth's ecosystems, is well demonstrated to decline under N input. However, it remains unclear why N fixers sustain N fixation in many forests under high atmospheric N deposition. To address this knowledge gap, we analyzed the response of the diazotroph community to low N loads (short-term and low N addition; 3-year N addition at the rates of 25-50 kg N ha-1 year-1) vs high loads (chronic and high N addition; 9-year N addition at the rate of 150 kg N ha-1 year-1) in forest soils using high-throughput sequencing. Rates of N fixation decreased under low and high N loads (by 13%-27% and 10%-12%, respectively). Richness and alpha diversity (ACE and Chao1) of the soil diazotroph community decreased under low but not high N loads. Approximately 67.1%-74.4% of the nifH gene sequences at the OTU level overlapped between the control and low N loads, but only 52.0%-53.6% of those overlapped between the control and high N loads, indicating a larger shift of diazotroph community composition under high N loads. Low N loads increased soil NH4+ concentrations, which decreased diazotroph community richness, diversity, and N fixation rates, whereas the increased soil NH4+ concentrations under high N loads did not have negative impacts on the structure and function of the diazotroph community. These findings indicate that diazotrophs sustain N fixation under high N deposition via adjustment of their community composition in forest soils. IMPORTANCE: This study examined the changes in soil diazotroph community under different loads of simulated N deposition and analyzed its relationship with N fixation rates in in five forests using high-throughput sequencing. The magnitudes of N fixation rates reduced by low N loads were higher than those by high N loads. Low N loads decreased richness and diversity of diazotroph community, whereas diazotroph community structure remained stable under high N loads. Compared with low N loads, high N loads resulted in a less similarity and overlap of nifH gene sequences among the treatments and a larger adjustment of diazotroph community. Low N loads increased soil NH4+ concentrations, which decreased diazotroph community richness, diversity, and N fixation rates, whereas the increased soil NH4+ under high N loads did not have negative impacts on diazotroph community structure and N fixation. Based on these findings, it is urgently needed to incorporate the loads of N deposition and the composition of diazotroph community into terrestrial N-cycling models for accurate understanding of N inputs in forest ecosystems.

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Nitrogen (N) addition can greatly influence soil inorganic phosphorus (Pi) and organic phosphorus (Po) transformations. However, whether and how the N compound forms may differentially affect the soil P fractions remain unclear. Here, we investigated the responses of soil Pi (labile Pi, moderately-occluded Pi, and recalcitrant Pi) and Po fractions (labile Po and stable Po) to varying addition rates of three N compounds ((NH4)2SO4, NH4NO3, and urea) in a meadow steppe in northern China. Our studies revealed that with increasing N addition rate, soil labile and moderately-occluded Pi increased, accompanied by decreases in soil recalcitrant Pi. This shift was attributed to N-induced soil acidification, which accelerated the conversion of recalcitrant Pi into labile and moderately-occluded Pi. Soil labile Po decreased with increasing rate of N addition, whilst soil stable Po was not affected. Regardless of the compound forms, N addition increased soil Olsen-P, suggesting a potential alleviation of P limitation in this grassland ecosystem. The effect of N addition on soil labile Pi was significantly greater with addition of urea than with addition of either (NH4)2SO4 or NH4NO3, indicating that urea was more efficient in enhancing soil P availability. Addition of (NH4)2SO4 imposed a more pronounced positive effect on soil moderately-occluded Pi than the addition of either NH4NO3 or urea, mainly due to the greater mobilization of recalcitrant Pi as a result of higher soil acidification strength of (NH4)2SO4. These findings underscore the importance of considering the distinct effects of different N compounds when studying grassland soil P dynamics and availability in response to N addition.

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Eutrophication resulting from anthropogenic activities has been recognized as a significant driver of changes in ecosystem functioning. Furthermore, it may exacerbate the top-down effect and thus exert an important impact on plant growth. To test this hypothesis, we conducted a 3-year manipulative field experiment to investigate the impacts of nitrogen addition and crab herbivory on the growth of Phragmites australis in the salt marsh of the Yellow River Delta. The results demonstrated that a 3-year nitrogen addition can significantly increase the total nitrogen and carbon content of P. australis leaves, thereby enhancing their nutritional value and palatability, as well as increasing the proportion of leaves consumed by crabs. Therefore, nitrogen addition together with crab herbivory had a significant negative effect on P. australis height, leaf length, and leaf breadth in the ambient crab and procedural crab cage treatment compared to the crab exclusion treatment. The structural equation modeling further substantiated these findings. The model revealed a direct and positive correlation between nitrogen addition and leaf nutrient content (path coefficient = 0.34). Additionally, it demonstrated a direct and positive relationship between leaf nutrient content and the proportion of leaves consumed by crabs (path coefficient = 0.22). Simultaneously, there was an observed negative correlation (path coefficient = - 0.37) between the proportion of leaves consumed by crabs and plant functional traits, represented by leaf length in the model, during 2018. Moreover, the crab exclusion treatment significantly reduced the proportion of leaves consumed by crabs and thus enhanced the P. australis individuals, leaf number, and biomass. Overall, crab herbivory had a significant detrimental top-down effect on the growth of P. australis, and nitrogen enrichment may exacerbate this top-down effect. The findings of our study highlight the combined adverse effects of nutrient enrichment and top-down on plant functional traits and plant growth. The findings of this study will contribute to a comprehensive understanding of the underlying factors influencing vegetation degradation in coastal wetland, thereby establishing a solid theoretical framework for the conservation and management of wetland ecosystems within the context of global environmental change.

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Soil respiration (Rs) is projected to be substantially affected by climate change, impacting the storage, equilibrium, and movement of terrestrial carbon (C). However, uncertainties surrounding the responses of Rs to climate change and soil nitrogen (N) enrichment are linked to mechanisms specific to diverse climate zones. A comprehensive meta-analysis was conducted to address this, evaluating the global effects of warming, increased precipitation, and N enrichment on Rs across various climate zones and ecosystems. Data from 123 studies, encompassing a total of 10,377 worldwide observations, were synthesized for this purpose. Annual Rs were modeled and their uncertainties were associated with a 1-km2 resolution global Rs database spanning from 1961 to 2022. Calibrating Rs using ensemble machine learning (EML) and employing 10-fold cross-validation, 13 environmental covariates were utilized. The meta-analysis findings revealed an upsurge in Rs rates in response to warming, with tropical, arid, and temperate climate zones exhibiting increases of 12 %, 13 %, and 16 %, respectively. Furthermore, increased precipitation led to stimulated Rs rates of 11 % and 9 % in tropical and temperate zones, respectively, while N deposition affected Rs in cold (+6 %) and tropical (+5 %) climate zones. The machine learning technique estimated the global soil respiration to range from 91 to 171 Pg C yr-1, with an average Rs of 700 ± 300 g C m-2 yr-1. The values ranged between 314 and 2500 g C m-2 yr-1, with the lowest and highest values observed in cold and tropical zones, respectively. Spatial variation in Rs was most pronounced in low-latitude areas, particularly in tropical rainforests and monsoon zones. Temperature, precipitation, and N deposition were identified as crucial environmental factors exerting significant influences on Rs rates worldwide. These factors underscore the interconnectedness between climate and ecosystem processes, therefore requiring explicit considerations of different climate zones when assessing responses of Rs to global change.

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Nitrogen (N) and carbon (C) inputs substantially affect soil microbial functions. However, the influences of long-term N and C additions on soil microbial resource limitation and heterotrophic respiration-fundamental microbial functional traits-remain unclear, impeding the understanding of how soil C dynamics respond to global change. In this study, the responses of soil microbial resource limitation and heterotrophic respiration (Rh) to 7-year N and biochar (BC) additions in a subtropical Moso bamboo (Phyllostachys edulis) plantation were investigated. We used eight treatments: Control, no N and BC addition; N30, 30 kg N (ammonium nitrate)·hm-2·a-1; N60, 60 kg N·hm-2·a-1; N90, 90 kg N·hm-2·a-1; BC20, 20 t BC (originating from Moso bamboo chips) hm-2; N30 + BC20, 30 kg N·hm-2·a-1 + 20 t BC hm-2; N60 + BC20, 60 kg N·hm-2·a-1 + 20 t BC hm-2; and N90 + BC20, 90 kg N·hm-2·a-1 + 20 t BC hm-2. Soil microbes were co-limited by N and phosphorus (P) and not limited by C in the control treatments. Long-term N addition enhanced soil microbial N and P limitation but significantly reduced soil Rh by 15.1 %-20.0 % relative to that in the control treatments. BC amendment alleviated soil microbial N and P limitation and significantly decreased C use efficiency by 10.9 %-42.1 % but increased Rh by 33.6 %-91.6 % in the long-term N-free and N-supplemented treatments (P < 0.05). Soil C- and N-acquisition enzyme activities were the dominant drivers of soil microbial resource limitation. Furthermore, microbial resource limitation was a more reliable predictor of Rh than soil resources or microbial biomass. The results suggested that long-term N and BC additions affect Rh by regulating microbial resource limitation, highlighting its significance in understanding soil C cycling under environmental change.

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Nitrogen (N) in the atmosphere frequently affects plant growth, ecological stoichiometric equilibrium, and homeostasis stability. However, the effect of N addition application on the growth of Hippophae rhamnoides seedlings remains ambiguous. We investigated the effects of N addition on the ecological stoichiometry and homeostatic characteristics of H. rhamnoides seedlings. Greenhouse cultivation experiments were conducted at five N application levels: 0 kg ha-1 yr-1(CK), 100 kg ha-1 yr-1 (N10), 200 kg ha-1 yr-1 (N20), 400 kg ha-1 yr-1 (N40), 800 kg ha-1 yr-1 (N80). The results showed that pH and available phosphorus (AP) significantly decreased with increasing N, whereas soil C:P and N:P ratios significantly increased under the N40 and N80 treatments. The leaf C:N ratio significantly decreased with increasing N, whereas the N:P ratio increased. With N addition, the C:N ratio of plant stems and roots significantly decreased, whereas the C:P and N:P ratios significantly increased. N addition was significantly correlated with the ecological stoichiometry of plant leaves and soil properties (0.38 and 0.84, respectively). Homeostasis of the organs of H. rhamnoides seedlings exhibited an absolute steady state. The C, N, and C:P ratios of the roots exhibited insensitive states under the N40 treatment. N addition significantly modified both the soil ecological stoichiometry and the stoichiometry of H. rhamnoides seedlings. However, it did not demonstrate a pronounced negative effect on the homeostasis of H. rhamnoides seedlings. This study offers new insights into the ecological adaptation process of H. rhamnoides, particularly concerning its nutrient distribution, utilization strategies, and stability.

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Biogenic volatile organic compounds (BVOCs) significantly impact atmospheric chemistry, with emissions potentially influenced by nitrogen (N) deposition. The response of BVOC emissions to increasing N deposition remains debated. In this study, we examined Eucalyptus urophylla (E. urophylla) using three N treatments: N0, N50, and N100 (0, 50, and 100 kg N hm-2 yr-1 N addition). These treatments were applied to mature E. urophylla trees in a plantation subjected to over 10 years of soil N addition in southern China, a region with severe N deposition. Seventeen BVOCs were measured, with isoprene (36.99 %), α-pinene (38.80 %), and d-limonene (14.27 %) being the predominant compounds under natural conditions. Total BVOC emissions under N50 were nearly double those under N0 and N100, with leaf net CO2 assimilation identified as the most critical photosynthetic parameter. Isoprene and α-pinene emissions significantly increased under N50 compared to N0, while d-limonene emission decreased under N100. Stronger correlations for individual BVOCs under N50 and N100 compared to N0 might be due to differences in BVOC biosynthetic pathways and storage structures. The localized canopy-scale emission factors (EFs) under N50 were significantly higher than the default values in the Model of Emissions of Gases and Aerosols from Nature (MEGAN), suggesting the model might underestimate BVOC emissions from Eucalyptus in southern China under increased N deposition. Additionally, the secondary pollutant formation potentials of BVOCs were evaluated, identifying isoprene and monoterpenes as primary precursors of ozone and secondary organic aerosols. This study provides insights into the impacts of increased N deposition on BVOC emissions and their contribution to secondary atmospheric pollution. Updating localized BVOC EFs for subtropical tree species in southern China is crucial to reduce uncertainties in BVOC estimations under current and future N deposition scenarios.

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Nitrogen application significantly affects microorganisms in agricultural ecosystems. However, it is still unclear how nitrogen application affects soil chemical properties and microbial communities in purple mudstone weathering products. In this study, a field soil column experiment was conducted in a typical purple soil area with four nitrogen fertilizer application gradients of 0 [CK], 280 [N1], 560 [N2], and 840 [N3] N kg ha-1. Nitrogen addition decreased the bacterial chao1 value and increased the bacterial evenness index. For both α- and ß-diversity, the effect of nitrogen addition on bacteria was much greater than that on fungi. Nitrogen addition significantly increased the relative abundance of Proteobacteria, Gemmatimonadetes, Bacteroidetes, and Ascomycota and decreased the relative abundance of Actinobacteria, Cyanobacteria, and Basidiomycota. Both pH and TC are the most important soil chemical properties influencing the bacterial and fungal communities. With the increases in the nitrogen application rate, the co-occurrence network complexity increased and then decreased. In summary, nitrogen fertilizer application could significantly change the soil chemical properties, microbial community diversity, composition, and co-occurrence network of purple mudstone weathering products. Among them, the N2 treatment (560 N kg∙ha-1) can more effectively stimulate the soil nutrients, enhance microbial network complexity, and promote further weathering of purple mudstone.

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Salt marsh has an important 'purification' role in coastal ecosystems by removing excess nitrogen that could otherwise harm aquatic life and reduce water quality. Recent studies suggest that salt marsh root exudates might be the 'control centre' for nitrogen transformation, but empirical evidence is lacking. Here we sought to estimate the direction and magnitude of nitrogen purification by salt marsh root exudates and gain a mechanistic understanding of the biogeochemical transformation pathway(s). To achieve this, we used a laboratory incubation to quantify both the root exudates and soil nitrogen purification rates, in addition to the enzyme activities and functional genes under Phragmites australis populations with different nitrogen forms addition (NO3-, NH4+ and urea). We found that NO3- and urea addition significantly stimulate P. australis root exudation of total acids, amino acids, total sugars and total organic carbon, while NH4+ addition only significantly increased total acids, amino acids and total phenol exudation. High total sugars, amino acids and total organic carbon concentrations enlarged nitrogen purification potential by stimulating the nitrogen purifying bacterial activities (including enzyme activities and related genes expression). Potential denitrification rates were not significantly elevated under NH4+ addition in comparison to NO3- and urea addition, which should be ascribed to total phenol self-toxicity and selective inhibition. Further, urea addition stimulated urease and protease activities with providing more NH4+ and NO2- substrates for elevated anaerobic ammonium oxidation rates among the nitrogen addition treatments. Overall, this study revealed that exogenous nitrogen could increase the nitrogen purification-associated bacterial activity through accelerating the root exudate release, which could stimulate the activity of nitrogen transformation, and then improve the nitrogen removal capacity in salt marsh.

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Atmospheric nitrogen (N) deposition in forests can affect soil microbial growth and turnover directly through increasing N availability and indirectly through altering plant-derived carbon (C) availability for microbes. This impacts microbial residues (i.e., amino sugars), a major component of soil organic carbon (SOC). Previous studies in forests have so far focused on the impact of understory N addition on microbes and microbial residues, but the effect of N deposition through plant canopy, the major pathway of N deposition in nature, has not been explicitly explored. In this study, we investigated whether and how the quantities (25 and 50 kg N ha-1 year-1) and modes (canopy and understory) of N addition affect soil microbial residues in a temperate broadleaf forest under 10-year N additions. Our results showed that N addition enhanced the concentrations of soil amino sugars and microbial residual C (MRC) but not their relative contributions to SOC, and this effect on amino sugars and MRC was closely related to the quantities and modes of N addition. In the topsoil, high-N addition significantly increased the concentrations of amino sugars and MRC, regardless of the N addition mode. In the subsoil, only canopy N addition positively affected amino sugars and MRC, implying that the indirect pathway via plants plays a more important role. Neither canopy nor understory N addition significantly affected soil microbial biomass (as represented by phospholipid fatty acids), community composition and activity, suggesting that enhanced microbial residues under N deposition likely stem from increased microbial turnover. These findings indicate that understory N addition may underestimate the impact of N deposition on microbial residues and SOC, highlighting that the processes of canopy N uptake and plant-derived C availability to microbes should be taken into consideration when predicting the impact of N deposition on the C sequestration in temperate forests.

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Background: Grass-legume mixture can effectively improve productivity and stimulate overyielding in artificial grasslands, but may be N-limited in semi-arid regions. This study investigated the effects of N addition on chlorophyll fluorescence and production in the grass-legume mixtures community. Methods: An N addition experiment was conducted in the Bothriochloa ischaemum and Lespedeza davurica mixture community, with seven mixture ratios (B0L10, B2L8, B4L6, B5L5, B6L4, B8L2, and B10L0) according to the sowing abundance of B.ischaemum and L.davurica and four N addition levels, N0, N25, N50, and N75 (0,25,50,75kgNhm-2 a-1), respectively. We analyzed the response of chlorophyll fluorescence parameters of the two species, the rapid light-response curves of chlorophyll fluorescence, as well as aboveground biomass (AGB) and overyielding. Results: Our results showed that the two species showed different photosynthetic strategies, with L.davurica having significantly higher initial fluorescence (Fo), effective photochemical quantum yield of PSII (ΦPSII), and coefficient of photochemical fluorescence quenching (qP) than B. ischaemum, consisting with results of rapid light-response curves. N addition and mixture ratio both had significant effects on chlorophyll fluorescence and AGB (p<0.001). The ΦPSII and qP of L.davurica were significantly lowest in B5L5 and B6L4 under N addition, and the effect of N varied with mixture ratio. The photosynthetic efficiency of B. ischaemum was higher in mixture than in monoculture (B10L0), and ΦPSII was significantly higher in N50 than in N25 and N50 at mixture communities except at B5L5. The community AGB was significantly higher in mixture communities than in two monocultures and highest at B6L4. In the same mixture ratio, the AGB was highest under the N50. The overyielding effects were significantly highest under the N75 and B6L4 treatments, mainly attributed to L.davurica. The partial least squares path models demonstrated that adding N increased soil nutrient content, and complementary utilization by B.ischaemum and L.davurica increased the photosynthetic efficiency. However, as the different photosynthetic strategies of these two species, the effect on AGB was offset, and the mixture ratio's effects were larger than N. Our results proposed the B6L4 and N50 treatments were the optimal combination, with the highest AGB and overyielding, moderate grass-legume ratio, optimal community structure, and forage values.

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A community functional structure may respond to environmental changes such as nitrogen (N) enrichment by altering intraspecific and interspecific trait variations. However, the relative contributions of both components in determining the community response to N enrichment are unclear. In this study, we measured the plant height (H), leaf area (LA), leaf dry matter content (LDMC), and specific leaf area (SLA) based on a nine-year N addition gradient experiment in an alpine meadow on the Tibetan Plateau. We examined the intraspecific and interspecific variations within and among the communities, the responses of traits in terms of community weighted mean (CWM) and non-weighted mean (CM) to N addition, and the effects of these trait variations on aboveground net primary productivity (ANPP). Our results show that N addition increased the interspecific variation in H while decreasing that of LA within the community, whereas it had no significant effects on the intraspecific variations in the four traits within the community. In contrast, N addition significantly increased the intraspecific variation in H and decreased that of LA among the communities. Moreover, the contribution of intraspecific variation was greater than that of the interspecific variation in terms of CWM for all traits, while the opposite contribution was observed in terms of CM, suggesting that the dominant species would have greater resilience while subdominant species would become less resistant to N addition. Further, intraspecific variations of LA and LDMC within the community played an important role in explaining community productivity. Our results highlight the importance of both intraspecific and interspecific variations in mediating functional trait responses to N enrichment, and intraspecific variation within the communities has important implications for community functioning that should be considered to better understand and predict the responses of the alpine grasslands to N enrichment.

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Introduction: The impact of nitrogen (N) deposition on the soil N-transforming process in grasslands necessitates further investigation into how N input influences the structural composition and diversity of soil N-cycling microbial communities across different grassland types. Methods: In this study, we selected two types of grassland soils in northwest Liaoning, temperate steppe and warm-temperate shrub, and conducted short-term N addition experiments using organic N, ammonium N, and nitrate N as sources with three concentration gradients to simulate N deposition. Illumina MiSeq sequencing technology was employed to sequence genes associated with N-cycling microbes including N-fixing, ammonia-oxidizing and denitrifying bacteria, and ammonia-oxidizing archaea. Results and discussion: The results revealed significant alterations in the structural composition and diversity of the N-cycling microbial community due to N addition, but the response of soil microorganisms varied inconsistent among different grassland types. Ammonium transformation rates had a greater impact on soils from temperate steppes while nitrification rates were more influential for soils from warm-temperate shrubs. Furthermore, the influence of the type of N source on soil N-cycling microorganisms outweighed that of its quantity applied. The ammonium type of nitrogen source is considered the most influential driving factor affecting changes in the structure of the microbial community involved in nitrogen transformation, while the amount of low nitrogen applied primarily determines the composition of soil bacterial communities engaged in nitrogen fixation and nitrification. Different groups of N-cycling microorganisms exhibited distinct responses to varying levels of nitrogen addition with a positive correlation observed between their composition, diversity, and environmental factors examined. Overall findings suggest that short-term nitrogen deposition may sustain dominant processes such as soil-N fixation within grasslands over an extended period without causing significant negative effects on northwestern Liaoning's grassland ecosystems within the next decade.

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Enhanced nitrogen (N) input is expected to influence the soil phosphorus (P) cycling through biotic and abiotic factors. Among these factors, soil microorganisms play a vital role in regulating soil P availability. However, the divergent contribution of functional microorganisms to soil P availability in the rhizosphere and bulk soil under N addition remains unclear. We conducted an N addition experiment with four N input rates (0, 5, 10, and 15 g N m-2 year-1) in an alpine meadow over three years. Metagenomics was employed to investigate the functional microbial traits in the rhizosphere and bulk soil. We showed that N addition had positive effects on microbial functional traits related to P-cycling in the bulk and rhizosphere soil. Specifically, high N addition significantly increased the abundance of most microbial genes in the bulk soil but only enhanced the abundance of five genes in the rhizosphere soil. The soil compartment, rather than the N addition treatment, was the dominant factor explaining the changes in the diversity and network of functional microorganisms. Furthermore, the abundance of functional microbial genes had a profound effect on soil available P, particularly in bulk soil P availability driven by the ppa and ppx genes, as well as rhizosphere soil P availability driven by the ugpE gene. Our results highlight that N addition stimulates the microbial potential for soil P mobilization in alpine meadows. Distinct microbial genes play vital roles in soil P availability in bulk and rhizosphere soil respectively. This indicates the necessity for models to further our knowledge of P mobilization processes from the bulk soil to the rhizosphere soil, allowing for more precise predictions of the effects of N enrichment on soil P cycling.

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Increasing nitrogen (N) input to coastal ecosystems poses a serious environmental threat. It is important to understand the responses and feedback of N removal microbial communities, particularly nitrifiers including the newly recognized complete ammonia-oxidizers (comammox), to improve aquaculture sustainability. In this study, we conducted a holistic evaluation of the functional communities responsible for nitrification by quantifying and sequencing the key functional genes of comammox Nitrospira-amoA, AOA-amoA, AOB-amoA and Nitrospira-nxrB in fish ponds with different fish feeding levels and evaluated the contribution of nitrifiers in the nitrification process through experiments of mixing pure cultures. We found that higher fish feeding dramatically increased N-related concentration, affecting the nitrifying communities. Compared to AOA and AOB, comammox Nitrospira and NOB were more sensitive to environmental changes. Unexpectedly, we detected an equivalent abundance of comammox Nitrospira and AOB and observed an increase in the proportion of clade A in comammox Nitrospira with the increase in fish feeding. Furthermore, a simplified network and shift of keystone species from NOB to comammox Nitrospira were observed in higher fish-feeding ponds. Random forest analysis suggested that the comammox Nitrospira community played a critical role in the nitrification of eutrophic aquaculture ponds (40-70 µM). Through the additional experiment of mixing nitrifying pure cultures, we found that comammox Nitrospira is the primary contributor to the nitrification process at 200 µM ammonium. These results advance our understanding of nitrifying communities and highlight the importance of comammox Nitrospira in driving nitrification in eutrophic aquaculture systems.

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The soil microbial carbon pump (MCP) is increasingly acknowledged as being directly linked to soil organic carbon (SOC) accumulation and stability. Given the close coupling of carbon (C) and nitrogen (N) cycles and the constraints imposed by their stoichiometry on microbial growth, N addition might affect microbial growth strategies with potential consequences for necromass formation and carbon stability. However, this topic remains largely unexplored. Based on two multi-level N fertilizer experiments over 10 years in two soils with contrasting soil fertility located in the North (Cambisol, carbon-poor) and Southwest (Luvisol, carbon-rich), we hypothesized that different resource demands of microorganism elicit a trade-off in microbial growth potential (Y-strategy) and resource-acquisition (A-strategy) in response to N addition, and consequently on necromass formation and soil carbon stability. We combined measurements of necromass metrics (MCP efficacy) and soil carbon stability (chemical composition and mineral associated organic carbon) with potential changes in microbial life history strategies (assessed via soil metagenomes and enzymatic activity analyses). The contribution of microbial necromass to SOC decreased with N addition in the Cambisol, but increased in the Luvisol. Soil microbial life strategies displayed two distinct responses in two soils after N amendment: shift toward A-strategy (Cambisol) or Y-strategy (Luvisol). These divergent responses are owing to the stoichiometric imbalance between microbial demands and resource availability for C and N, which presented very distinct patterns in the two soils. The partial correlation analysis further confirmed that high N addition aggravated stoichiometric carbon demand, shifting the microbial community strategy toward resource-acquisition which reduced carbon stability in Cambisol. In contrast, the microbial Y-strategy had the positive direct effect on MCP efficacy in Luvisol, which greatly enhanced carbon stability. Such findings provide mechanistic insights into the stoichiometric regulation of MCP efficacy, and how this is mediated by site-specific trade-offs in microbial life strategies, which contribute to improving our comprehension of soil microbial C sequestration and potential optimization of agricultural N management.

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Tall clonal grasses commonly display competitive advantages with nitrogen (N) enrichment. However, it is currently unknown whether the height is derived from the vegetative or reproductive module. Moreover, it is unclear whether the height of the vegetative or reproductive system regulates the probability of extinction and colonization, and determines species diversity. In this study, the impacts on clonal grasses were studied in a field experiment employing two frequencies (twice a year vs. monthly) crossing with nine N addition rates in a temperate grassland, China. We found that the N addition decreased species frequency and increased extinction probability, but did not change the species colonization probability. A low frequency of N addition decreased species frequency and colonization probability, but increased extinction probability. Moreover, we found that species reproductive height was the best index to predict the extinction probability of clonal grasses in N-enriched conditions. The low frequency of N addition may overestimate the negative effect from N deposition on clonal grass diversity, suggesting that a higher frequency of N addition is more suitable in assessing the ecological effects of N deposition. Overall, this study illustrates that reproductive height was associated with the clonal species extinction probability under N-enriched environment.

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Atmospheric nitrogen deposition can alter soil microbial communities and further impact the structure and function of forest ecosystems. However, most studies are focused on positive or negative effects after nitrogen addition, and few studies pay attention to its interruption. In order to investigate whether interruption after different levels of short-term N additions still benefit soil health, we conducted a 2-year interruption after a 4-year short-term nitrogen addition (10 and 20 kg N·hm-2·yr-1) experiment; then, we compared soil microbial diversity and structure and analyzed soil physicochemical properties and their correlations before and after the interruption in Larix olgensis forest soil in northeast China. The results showed that soil ecological stabilization of Larix olgensis forest further improved after the interruption compared to pre-interruption. The TN, C:P, N:P, and C:N:P ratios increased significantly regardless of the previous nitrogen addition concentration, and soil nutrient cycling was further promoted. The relative abundance of the original beneficial microbial taxa Gemmatimonas, Sphingomonas, and Pseudolabrys increased; new beneficial bacteria Ellin6067, Massilia, Solirubrobacter, and Bradyrhizobium appeared, and the species of beneficial soil microorganisms were further improved. The results of this study elucidated the dynamics of the bacterial community before and after the interruption of short-term nitrogen addition and could provide data support and a reference basis for forest ecosystem restoration strategies and management under the background of global nitrogen deposition.

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Nitrogen (N) deposition is a global environmental issue that can have significant impacts on the community structure and function in ecosystems. Fungi play a key role in soil biogeochemical cycles and their community structures are tightly linked to the health and productivity of forest ecosystems. Based on high-throughput sequencing and ergosterol extraction, we examined the changes in community structure, composition, and biomass of soil ectomycorrhizal (ECM) and saprophytic (SAP) fungi in 0-10 cm soil layer after 8 years of continuous N addition and their driving factors in a temperate Korean pine plantation in northeast China. Our results showed that N addition increased fungal community richness, with the highest richness and Chao1 index under the low N treatment (LN: 20 kg N ha-1 yr-1). Based on the FUN Guild database, we found that the relative abundance of ECM and SAP fungi increased first and then decreased with increasing N deposition concentration. The molecular ecological network analysis showed that the interaction between ECM and SAP fungi was enhanced by N addition, and the interaction was mainly positive in the ECM fungal network. N addition increased fungal biomass, and the total fungal biomass (TFB) was the highest under the MN treatment (6.05 ± 0.3 mg g-1). Overall, we concluded that N addition changed soil biochemical parameters, increased fungal activity, and enhanced functional fungal interactions in the Korean pine plantation over an 8-year simulated N addition. We need to consider the effects of complex soil conditions on soil fungi and emphasize the importance of regulating soil fungal community structure and biomass for managing forest ecosystems. These findings could deepen our understanding of the effects of increased N deposition on soil fungi in temperate forests in northern China, which can provide the theoretical basis for reducing the effects of increased N deposition on forest soil.

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Soil nitrogen (N) availability affects plant carbon (C) utilization. However, it is unclear how various tree functional types respond to N addition in terms of C assimilation, allocation, and storage. Here, a microcosm experiment with dual 13C and 15N labeling was conducted to study the effects of N addition (i.e., control, 0 g N kg-1; moderate N addition, 1.68 g N kg-1; and high N addition, 3.36 g N kg-1 soil) on morphological traits, on changes in nonstructural carbohydrates (NSC) in different organs, as well as on C and N uptake and allocation in three European temperate forest tree species (i.e., Acer pseudoplatanus, Picea abies and Abies alba). Our results demonstrated that root N uptake rates of the three tree species increased by N addition. In A. pseudoplatanus, N uptake by roots, N allocation to aboveground organs, and aboveground biomass allocation significantly improved by moderate and high N addition. In A. alba, only the high N addition treatment considerably raised aboveground N and C allocation. In contrast, biomass as well as C and N allocation between above and belowground tissues were not altered by N addition in P. abies. Meanwhile, NSC content as well as C and N coupling (represented by the ratio of relative 13C and 15N allocation rates in organs) were affected by N addition in A. pseudoplantanus and P. abies but not in A. alba. Overall, A. pseudoplatanus displayed the highest sensitivity to N addition and the highest N requirement among the three species, while P. abies had a lower N demand than A. alba. Our findings highlight that the responses of C and N allocation to soil N availability are species-specific and vary with the amount of N addition.

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