N-cycling Microbial Communities Research Articles

Groundwater flow regime shapes nitrogen functional traits by affecting microbial community assembly processes in the subsurface

The complex nitrogen (N) cycle in groundwater systems is affected by both biological and environmental factors. The interactions between hydrogeological conditions and the microbial community assembly processes that impact N-cycling processes remain poorly understood. We explored the assembly patterns of N-cycling microbial communities along the groundwater flow path. The environmental heterogeneity in different hydrological phases increased along the flow path (mean Ed: 0.16–0.49), accompanied by different microbial community assembly patterns. The assembly patterns that engaged in dissimilatory nitrate reduction to ammonium (DNRA) and denitrification changed across the water-sediment phases. Nitrifying microorganisms in the discharge area were mainly influenced by heterogeneous selection (41–69 %), and were closely correlated with dissolved oxygen (DO) concentrations. Homogeneity along flow-through increased stochastic assemblies, such as downstream drift of anammox bacterial (AnAOB) communities. Thus, the N removal pathway changed from “nitrification-denitrification” in the recharge area to “partial nitrification-anammox” in the discharge area. The increasing environmental heterogeneity brought more deterministic assembly patterns of N-cycling communities, linked to higher community turnover along the groundwater flow path. This study indicated that groundwater flow regime determined microbial community assembly patterns, providing valuable insight into the response of N transitions to environmental variations in groundwater systems.

Mineral and organic fertilisation influence ammonia oxidisers and denitrifiers and nitrous oxide emissions in a long-term tillage experiment

Nitrous oxide (N2O) emissions from different agricultural systems have been studied extensively to understand the mechanisms underlying their formation. While a number of long-term field experiments have focused on individual agricultural practices in relation to N2O emissions, studies on the combined effects of multiple practices are lacking. This study evaluated the effect of different tillage [no-till (NT) vs. conventional plough tillage (CT)] in combination with fertilisation [mineral (MIN), compost (ORG), and unfertilised control (CON)] on seasonal N2O emissions and the underlying N-cycling microbial community in one maize growing season. Rainfall events after fertilisation, which resulted in increased soil water content, were the main triggers of the observed N2O emission peaks. The highest cumulative emissions were measured in MIN fertilisation, followed by ORG and CON fertilisation. In the period after the first fertilisation CT resulted in higher cumulative emissions than NT, while no significant effect of tillage was observed cumulatively across the entire season. A higher genetic potential for N2O emissions was observed under NT than CT, as indicated by an increased (nirK + nirS)/(nosZI + nosZII) ratio. The mentioned ratio under NT decreased in the order CON > MIN > ORG, indicating a higher N2O consumption potential in the NT-ORG treatment, which was confirmed in terms of cumulative emissions. The AOB/16S ratio was strongly affected by fertilisation and was higher in the MIN than in the ORG and CON treatments, regardless of the tillage system. Multiple regression has revealed that this ratio is one of the most important variables explaining cumulative N2O emissions, possibly reflecting the role of bacterial ammonia oxidisers in minerally fertilised soil. Although the AOB/16S ratio aligned well with the measured N2O emissions in our experimental field, the higher genetic potential for denitrification expressed by the (nirK + nirS)/(nosZI + nosZII) ratio in NT than CT was not realized in the form of increased emissions. Our results suggest that organic fertilisation in combination with NT shows a promising combination for mitigating N2O emissions; however, addressing the yield gap is necessary before incorporating it in recommendations for farmers.

Carbohydrate addition strategy affects nitrogen dynamics, budget and utilization, and its microbial mechanisms in biofloc-based Penaeus vannamei culture

Biofloc technology (BFT) is a microbial biotechnology that has shown great potential in water quality control and nutrient re-utilization towards sustainable aquaculture. Carbohydrate addition is the main operation means for the application of BFT; however, we are still unclear how it affects microbial-driven nitrogen (N) transformation and utilization in BFT systems. In this study, autotrophic- and heterotrophic-dominated BFT systems (AS vs. HS) in triplicates were operated by two molasses addition strategies for a 13-week production trial of P. vannamei. Throughout the trial, more amount of bioflocs (in terms of VSS) were produced in HS than in AS. Higher concentrations of NH4+-N, NO2−-N, NO3−-N, TIN and TN while lower concentrations of TON were monitored in AS compared to HS. N budget analysis showed that the harvested shrimp deposited >34.4% of input N; and higher N retention in the final culture water while lower N loss and N discharge from the systems were found in AS compared to HS. Higher efficiencies of gross and feed N utilization were achieved in HS (36.6% and 38.8%) than in AS (34.5% and 34.9%). Metagenomic analysis revealed that abundant and diverse N-cycling microbial communities and functional genes in bioflocs constituted a complete and complex N-cycling network, which was the driving force of N transformation and recycling in the culture water of systems. The lower abundances of nitrifying bacteria and associated genes led to the inhibition of nitrification under high molasses addition in HS, which accounted for the differences in N dynamics, budget and utilization efficiency between AS and HS. Together, a quantitative model of N migration and transformation driven by N-cycling microbial communities of bioflocs was depicted in closed BFT systems for shrimp aquaculture. This study firstly revealed the N-cycling microbiome and functional effects in BFT systems, which provides theoretical and technical foundations for efficient N management in intensive aquaculture.

Nitrogen-cycling microbial communities respond differently to nitrogen addition under two contrasting grassland soil types.

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. 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. 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.

氮磷添加下典型温带、亚热带森林土壤氮循环功能基因丰度和微生物群落属性数据集

Nitrogen (N) and phosphorus (P) are essential nutrients for soil microbial growth and activities. There are notable differences in the concentrations of N and P and the characteristics of microbial communities observed between temperate and subtropical forests soils, and this is also true for soil vertical profiles. As a consequence of the rise in global atmospheric N deposition, soil nutrient imbalances and P limitation are increasingly aggravated in eastern China. However, there is a lack of systematic dataset showing how N deposition and P addition affect soil N cycling microbes and soil physicochemical properties in eastern China; hence a comprehensive dataset is needed to a better understanding of the impact of N deposition and P addition on soil nitrogen cycle. This dataset is constructed based on the field experimental setup in the forests of Chinese Ecosystem Research Network (CERN). To obtain a systematic dataset on N-cycling microbial communities, we conducted measurements and collected data of soil N-cycling functional genes abundance, N-cycling microbial community diversities and compositions and enzyme activities in three typical forests with N and/or P additions. We collected 0-10 cm top soils from a temperate forest (Changbai Mountain) and two subtropical forests (Qianyanzhou and Dinghu Mountain) with field N and/or P additions, and the soils along 0-80 cm vertical profiles without nutrient additions. The N-cycling functional genes involved in the datasets cover ammonia oxidizing archaea amoA, ammonia oxidizing bacteria amoA, nirK, nirS, nosZ, qnorB, narG, nir, nifH and chiA. The potential activities included in the datasets cover nitrification activity, denitrification activity, nitrogen fixation activity and organic nitrogen decomposition enzyme activity. The datasets also include soil physiochemical properties, including soil pH, moisture content, N2O emission, net nitrification rate and the concentrations of organic carbon, NH4+ , NO3− , available phosphorus, total nitrogen, total carbon, total phosphorus and dissolved organic carbon. The creation and sharing of the datasets can provide data and support for understanding the microbial mechanisms of soil N-cycling and the modelling processes under the scenario of aggravated N deposition and P addition. The datasets will also provide support for forest management efforts with regard to greenhouse gas N2O emission, N and nutrient loss in forest ecosystems.

Straw return of maize and soybean enhances soil biological nitrogen fixation by altering the N-cycling microbial community

Crop straw return can enhance soil biological nitrogen (N) fixation (BNF) by improving soil properties, but the impacts of several crop straw mixture return on BNF and the N-cycling microbial community and their relationships are still unclear. Here, we investigated BNF and N-cycling microbial communities in soil with maize straw and maize-soybean straw mixtures, respectively. The results showed that soil BNF was highest (2.51 μg g−1 DW) in the 10 mg g−1 straw mixture treatment, being 25.1-fold and 250-fold higher than that in the 10 mg g−1 maize straw treatment and control, respectively. The copy numbers of nifH, nirS and nosZ significantly increased, and the relative abundances of the dominant microbial communities carrying the above genes were remarkably improved by 10 mg g−1 straw mixture return. The increase in soil BNF after straw return was significantly associated not only with N-fixing microbial communities but also with other N transformation microbial communities. In addition, 7, 8, 11 and 5 genera for the chiA, nirK, nirS and nosZ microbial communities, respectively, were positively associated with BNF. Therefore, straw return affected the expression of the functional genes and microbial communities involved in soil N-cycling by altering the soil properties (soil organic matter, pH, NO3− and available potassium), which ultimately enhanced the soil N fixation rate. Moreover, soil properties and N-cycling functional microbial communities have the largest common explanation (58.6%) for the differences in BNF. Our findings contribute to a better understanding of the mechanistic link between N-cycling microbial communities and BNF improvement after straw return, which provides theoretical guidance for the sustainable development of agriculture.

Long-Term Pollution Does Not Inhibit Denitrification and DNRA by Adapted Benthic Microbial Communities

Denitrification in sediments is a key microbial process that removes excess fixed nitrogen, while dissimilatory nitrate reduction to ammonium (DNRA) converts nitrate to ammonium. Although microorganisms are responsible for essential nitrogen (N) cycling, it is not yet fully understood how these microbially mediated processes respond to toxic hydrophobic organic compounds (HOCs) and metals. In this study, we sampled long-term polluted sediment from the outer harbor of Oskarshamn (Baltic Sea), measured denitrification and DNRA rates, and analyzed taxonomic structure and N-cycling genes of microbial communities using metagenomics. Results showed that denitrification and DNRA rates were within the range of a national reference site and other unpolluted sites in the Baltic Sea, indicating that long-term pollution did not significantly affect these processes. Furthermore, our results indicate an adaptation to metal pollution by the N-cycling microbial community. These findings suggest that denitrification and DNRA rates are affected more by eutrophication and organic enrichment than by historic pollution of metals and organic contaminants.

Insight into the microbial nitrogen cycle in riparian soils in an agricultural region

Riparian zones are considered as an effective measure on preventing agricultural non-point source nitrogen (N) pollution. However, the mechanism underlying microbial N removal and the characteristics of N-cycle in riparian soils remain elusive. In this study, we systematically monitored the soil potential nitrification rate (PNR), denitrification potential (DP), as well as net N2O production rate, and further used metagenomic sequencing to elucidate the mechanism underlying microbial N removal. As a whole, the riparian soil had a very strong denitrification, with the DP 3.17 times higher than the PNR and 13.82 times higher than the net N2O production rate. This was closely related to the high soil NO3−-N content. In different profiles, due to the influence of extensive agricultural activities, the soil DP, PNR, and net N2O production rate near the farmland edge were relatively low. In terms of N-cycling microbial community composition, the taxa of denitrification, dissimilatory nitrate reduction, and assimilatory nitrate reduction accounted for a large proportion, all related to NO3−-N reduction. The N-cycling microbial community in waterside zone showed obvious differences to the landside zone. The abundances of N-fixation and anammox genes were significantly higher in the waterside zone, while the abundances of nitrification (amoA&B&C) and urease genes were significantly higher in the landside zone. Furthermore, the groundwater table was an important biogeochemical hotspot in the waterside zone, the abundance of N-cycle genes near the groundwater table was at a relative higher level. In addition, compared to different soil depths, greater variation in N-cycling microbial community composition was observed between different profiles. These results reveal some characteristics of the soil microbial N-cycle in the riparian zone in an agricultural region and are helpful for restoration and management of the riparian zone.

Metagenomics reveals elevated temperature causes nitrogen accumulation mainly by inhibiting nitrate reduction process in polluted water

Determining the response of functional genes and microbiota involved in the nitrogen (N) cycle to warming in the face of global climate change is a hotpot topic. However, whether and how elevated temperature affects the N-cycle genes in polluted water remains unclear. Based on metagenomics, we investigated the responses of the whole N-cycling genes and their microbial communities to the temperature gradients (23, 26, 29, 32, and 35 °C) using animal cadavers as an N-pollution model. We found that the abundance of gene families involved in glutamate metabolism, assimilatory nitrate reduction to nitrite (ANRN), and denitrification pathways decreased with temperature. Moreover, warming reduced the diversity of N-cycling microbial communities. Ecological network analysis indicated that elevated temperature intensified the mutual competition of N-cycle genes. The partial least squares path model (PLS-PM) showed that warming directly suppressed most N-cycle pathways, especially glutamate metabolism, denitrification, and ANRN pathways. Corpse decay also indirectly inhibited N-cycling via regulating N content and microbial communities. Our results highlight warming leads to N accumulation by inhibiting the ANRN and denitrification pathways, which may jeopardize ecological environment security. Our study is expected to provide valuable insights into the complex N-cycle process and N-pollution in warmer aquatic ecosystems.

Nitrogen species and microbial community coevolution along groundwater flowpath in the southwest of Poyang Lake area, China

Nitrate and ammonia overload in groundwater can lead to eutrophication of surface water in areas where surface water is recharged by groundwater. However, this process remained elusive due to the complicated groundwater N cycling, which is governed by the co-evolution of hydrogeochemical conditions and N-cycling microbial communities. Herein, this process was studied along a generalized groundwater flowpath in Ganjing Delta, Poyang Lake area, China. From groundwater recharge to the discharge area near the lake, oxidation-reduction potential (ORP), NO3–N, and NO2–N decreased progressively, while NH3–N, total organic carbon (TOC), Fe2+, sulfide, and TOC/NO3− ratio accumulated in the lakeside samples. The anthropogenic influences such as sewage and agricultural activities drove the nitrate distribution, as observed by Cl− vs. NO3−/Cl− ratio and isotopic composition of nitrate. The hydrogeochemical evolution was intimately coupled with the changes in microbial communities. Variations in microbial community structures was significantly explained by Fe2+, NH3–N, and sulfide, while TOC/NO3− controlled the distribution of predicted N cycling gene. The absence of NH3–N in groundwater upstream was accompanied by the enrichment in Acinetobacter capable of nitrification and aerobic denitrification. These two processes were also supported by Ca2+ + Mg2+ vs. HCO3− ratio and isotopic composition of NO3−. The DNRA process downstream was revealed by both the presence of DNRA-capable microbes such as Arthrobacter and the isotopic composition of NH4+ in environments with high concentrations of NH3–N, TOC/NO3−, Fe2+, and sulfide. This coupled evolution of N cycling and microbial community sheds new light on the N biogeochemical cycle in areas where surface water is recharged by groundwater.

Soil Microbial Community Involved in Nitrogen Cycling in Rice Fields Treated with Antiozonant under Ambient Ozone.

Ethylenediurea (EDU) can effectively mitigate the crop yield loss caused by ozone (O3), a major, phytotoxic air pollutant. However, the relevant mechanisms are poorly understood, and the effect of EDU on soil ecosystems has not been comprehensively examined. In this study, a hybrid rice variety (Shenyou 63) was cultivated under ambient O3 and sprayed with 450 ppm EDU or water every 10 days. Real time quantitative polymerase chain reaction (RT-qPCR) showed that EDU had no significant effect on the microbial abundance in either rhizospheric or bulk soils. By applying both metagenomic sequencing and the direct assembly of nitrogen (N)-cycling genes, EDU was found to decrease the abundance of functional genes related to nitrification and denitrification processes. Moreover, EDU increased the abundance of genes involved in N-fixing. Although the abundance of some functional genes did not change significantly, nonmetric multidimensional scaling (NMDS) and a principal coordinates analysis (PCoA) suggested that the microbial community structure involved in N cycling was altered by EDU. The relative abundances of nifH-and norB-harboring microbial genera in the rhizosphere responded differently to EDU, suggesting the existence of functional redundancy, which may play a key role in sustaining microbially mediated N-cycling under ambient O3. IMPORTANCE Ethylenediurea (EDU) is hitherto the most efficient phytoprotectant agent against O3 stress. However, the underlying biological mechanisms of its mode of action are not clear, and the effects of EDU on the environment are still unknown, limiting its large-scale application in agriculture. Due to its sensitivity to environmental changes, the microbial community can be used as an indicator to assess the environmental impacts of agricultural practices on soil quality. This study aimed to unravel the effects of EDU spray on the abundance, community structure, and ecological functions of microbial communities in the rhizosphere of rice plants. Our study provides a deep insight into the impact of EDU spray on microbial-mediated N cycling and the structure of N-cycling microbial communities. Our findings help to elucidate the mode of action of EDU in alleviating O3 stress in crops from the perspective of regulating the structure and function of the rhizospheric soil microbial community.

Combined application of organic manure and chemical fertilizers stabilizes soil N-cycling microflora

• The abundance of N-cycling genes differently responded to NPK application.• Chemical NPK application greatly altered the N-cycling microbial community structure.• Soil acidification was the main driver for the variation in the N-cycling microbial community.• Manure addition was beneficial for stabilizing the N-cycling microbial community.Straw and manure are widely applied to agricultural systems, and greatly shape soil N-cycling microflora. However, we still lack a comprehensive understanding of how these organic materials structure soil N-cycling microbial communities. In this study, metagenomic analysis was performed to investigate the compositional variation in N-cycling microbial communities in a 30-year long-term experiment under five fertilization regimes: no fertilization (Control), chemical fertilization only (NPK), and NPK with wheat straw (NPK + HS), pig manure (NPK + PM), and cow manure (NPK + CM). Long-term NPK application differentially changed N-cycling gene abundance and greatly altered N-cycling microbial community structure. NPK + HS resulted in a similar pattern to NPK in terms of gene abundance and community structure. However, NPK + PM and NPK + CM significantly increased most genes and resulted in a community similar to that of the Control. Further analysis revealed that serious soil acidification caused by long-term NPK fertilization was a major factor for the variation in N-cycling microbial communities. The addition of alkaline manure, rather than wheat straw, stabilized the N-cycling microbial community structure presumably by alleviating soil acidification. These results revealed the strong impact of soil acidification on microbial N-cycling communities and illustrated the possibility of resolving nitrogen-related environmental problems by manipulating pH in acidified agricultural soils.

Conversion of coastal wetland to aquaculture ponds decreased N2O emission: Evidence from a multi-year field study

Land reclamation is a major threat to the world's coastal wetlands, and it may influence the biogeochemical cycling of nitrogen in coastal regions. Conversion of coastal marshes into aquaculture ponds is common in the Asian Pacific region, but its impacts on the production and emission of nitrogen greenhouse gases remain poorly understood. In this study, we compared N2O emission from a brackish marsh and converted shrimp aquaculture ponds in the Shanyutan wetland, the Min River Estuary in Southeast China over a three-year period. We also measured sediment and porewater properties, relevant functional gene abundance, sediment N2O production potential and denitrification potential in the two habitats. Results indicated that the pond sediment had lower N-substrate availability, lower ammonia oxidation (AOA and comammox Nitrospira amoA), nitrite reduction (nirK and nirS) and nitrous oxide reduction (nosZ Ⅰ and nosZ Ⅱ) gene abundance and lower N2O production and denitrification potentials than in marsh sediments. Consequently, N2O emission fluxes from the aquaculture ponds (range 5.4–251.8 μg m–2 h–1) were significantly lower than those from the marsh (12.6–570.7 μg m–2 h–1). Overall, our results show that conversion from marsh to shrimp aquaculture ponds in the Shanyutan wetland may have diminished nutrient input from the catchment, impacted the N-cycling microbial community and lowered N2O production capacity of the sediment, leading to lower N2O emissions. Better post-harvesting management of pond water and sediment may further mitigate N2O emissions caused by the aquaculture operation.

The active functional microbes contribute differently to soil nitrification and denitrification potential under long-term fertilizer regimes in North-East China.

Nitrogen (N) cycling microorganisms mediate soil nitrogen transformation processes, thereby affecting agricultural production and environment quality. However, it is not fully understood how active N-cycling microbial community in soil respond to long-term fertilization, as well as which microorganisms regulate soil nitrogen cycling in agricultural ecosystem. Here, we collected the soils from different depths and seasons at a 29-year fertilization experimental field (organic/chemical fertilizer), and investigated the transcriptions of N-cycling functional genes and their contribution to potential nitrification and denitrification. We found that long-term fertilization exerted significant impacts on the transcript abundances of nitrifiers (AOA amoA, AOB amoA and hao) and denitrifiers (narG and nosZ), which was also notably influenced by season variation. The transcriptions of AOA amoA, hao, and narG genes were lowest in autumn, and AOB amoA and nosZ transcript abundances were highest in autumn. Compared to no fertilization, soil potential nitrification rate (PNR) was reduced in fertilization treatments, while soil potential denitrification rate (PDR) was significantly enhanced in organic combined chemical fertilizer treatment. Both PNR and PDR were highest in 0–20 cm among the tested soil depths. Path model indicated active nitrifiers and denitrifiers had significant impact on soil PNR and PDR, respectively. The transcriptions of AOA amoA and nxr genes were significantly correlated with soil PNR (Pearson correlation, r > 0.174, p < 0.05). Significant correlation of napA and nosZ transcriptions with soil PDR (Pearson correlation, r > 0.234, p < 0.05) was also revealed. Random forest analysis showed that SOC content and soil pH were the important factors explaining the total variance of active nitrifers and denitrifiers, respectively. Taken together, long-term fertilization regimes reduced soil PNR and enhanced PDR, which could be attributed to the different responses of active N-cycling microorganisms to soil environment variations. This work provides new insight into the nitrogen cycle, particularly microbial indicators in nitrification and denitrification of long-term fertilized agricultural ecosystems.

Urea fertilization and grass species alter microbial nitrogen cycling capacity and activity in a C4 native grassland.

Soil microbial transformation of nitrogen (N) in nutrient-limited native C4 grasslands can be affected by N fertilization rate and C4 grass species. Here, we report in situ dynamics of the population size (gene copy abundances) and activity (transcript copy abundances) of five functional genes involved in soil N cycling (nifH, bacterial amoA, nirK, nirS, and nosZ) in a field experiment with two C4 grass species (switchgrass (Panicum virgatum) and big bluestem (Andropogon gerardii)) under three N fertilization rates (0, 67, and 202 kg N ha−1). Diazotroph (nifH) abundance and activity were not affected by N fertilization rate nor grass species. However, moderate and high N fertilization promoted population size and activity of ammonia oxidizing bacteria (AOB, quantified via amoA genes and transcripts) and nitrification potential. Moderate N fertilization increased abundances of nitrite-reducing bacterial genes (nirK and nirS) under switchgrass but decreased these genes under big bluestem. The activity of nitrous oxide reducing bacteria (nosZ transcripts) was also promoted by moderate N fertilization. In general, high N fertilization had a negative effect on N-cycling populations compared to moderate N addition. Compared to big bluestem, the soils planted with switchgrass had a greater population size of AOB and nitrite reducers. The significant interaction effects of sampling season, grass species, and N fertilization rate on N-cycling microbial community at genetic-level rather than transcriptional-level suggested the activity of N-cycling microbial communities may be driven by more complex environmental factors in native C4 grass systems, such as climatic and edaphic factors.

Limited Impacts of Cover Cropping on Soil N-Cycling Microbial Communities of Long-Term Corn Monocultures.

Cover cropping (CC) is a promising in-field practice to mitigate soil health degradation and nitrogen (N) losses from excessive N fertilization. Soil N-cycling microbial communities are the fundamental drivers of these processes, but how they respond to CC under field conditions is poorly documented for typical agricultural systems. Our objective was to investigate this relationship for a long-term (36 years) corn [Zea mays L.] monocultures under three N fertilizer rates (N0, N202, and N269; kg N/ha), where a mixture of cereal rye [Secale cereale L.] and hairy vetch [Vicia villosa Roth.] was introduced for two consecutive years, using winter fallows as controls (BF). A 3 × 2 split-plot arrangement of N rates and CC treatments in a randomized complete block design with three replications was deployed. Soil chemical and physical properties and potential nitrification (PNR) and denitrification (PDR) rates were measured along with functional genes, including nifH, archaeal and bacterial amoA, nirK, nirS, and nosZ-I, sequenced in Illumina MiSeq system and quantified in high-throughput quantitative polymerase chain reaction (qPCR). The abundances of nifH, archaeal amoA, and nirS decreased with N fertilization (by 7.9, 4.8, and 38.9 times, respectively), and correlated positively with soil pH. Bacterial amoA increased by 2.4 times with CC within N269 and correlated positively with soil nitrate. CC increased the abundance of nirK by 1.5 times when fertilized. For both bacterial amoA and nirK, N202 and N269 did not differ from N0 within BF. Treatments had no significant effects on nosZ-I. The reported changes did not translate into differences in functionality as PNR and PDR did not respond to treatments. These results suggested that N fertilization disrupts the soil N-cycling communities of this system primarily through soil acidification and high nutrient availability. Two years of CC may not be enough to change the N-cycling communities that adapted to decades of disruption from N fertilization in corn monoculture. This is valuable primary information to understand the potentials and limitations of CC when introduced into long-term agricultural systems.

Arbuscular mycorrhizal fungi and plant diversity drive restoration of nitrogen-cycling microbial communities.

Soil microbial communities, key players of many crucial ecosystem functions, are susceptible to environmental disturbances, which might cause the loss of microbial diversity and functions. However, few ecological concepts and practices have been developed for rescuing stressed soil microbial communities. Here, we manipulated an experiment with or without arbuscular mycorrhizal fungi (AMF) inoculation and at three levels (one, three and six species) of plant diversity to disentangle how the AMF and vegetation rescue soil nitrogen (N) -cycling microbial loop from simulated degraded soil ecosystem. Our results showed that AMF inoculation improved the restoration of soil N-cycling microbial communities. This improved restoration was related to the role of AMF in enhancing interactions within the N-cycling microbial loop. Furthermore, increased plant diversity strengthened the role of AMF in rescuing N-cycling microbial communities. Our findings provide novel insights into the roles of AMF and plant diversity in facilitating the rescue of microbial communities in degraded terrestrial ecosystems.

Soil and temperature effects on nitrification and denitrification modified N2O mitigation by 3,4-dimethylpyrazole phosphate

Nitrification inhibitors have often been found to reduce nitrous oxide (N2O) emissions from agricultural soils, but N2O mitigation is variable and may depend on soil and climate variables. This study investigated how key abiotic factors modified the effect of Vizura®, a new formulation with the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) for livestock slurry, on N2O emissions. Two different incubations were carried out to investigate the main and interactive effects of selected factors on N2O emissions after manure application to soil. Experiment 1 studied two contrasting soil textures (coarse sandy vs sandy loam soil) and temperatures (10 vs 20 °C); Experiment 2 used the coarse sandy soil to compare two levels of soil pH (4.9 vs 6.4) and nitrate availability (0 vs 50 mg N kg−1). Nitrous oxide emissions and spatial distribution of mineral N were determined in both experiments, while temporal dynamics of mineral N pools and N-cycling microbial communities were investigated in Experiment 2 only. In sandy loam soil, the efficiency of DMPP to mitigate N2O emissions after manure application was similar at 10 and 20 °C, while in the coarse sandy soil DMPP was not effective at 10 °C, possibly because of low nitrification activity. Nitrous oxide emissions from the coarse sandy soil were higher at pH 4.9 than at pH 6.4, which was accompanied by lower abundance of nosZ genes and therefore potentially higher N2O/(N2+N2O) product ratios. DMPP significantly reduced the abundance of ammonia-oxidizing bacteria (AOB) on all sampling days, but had no effect on ammonia-oxidizing archaea (AOA). The inhibitor also reduced the abundance of comammox, which are newly discovered complete ammonia oxidizers. This study showed how soil conditions (soil texture, temperature, pH), through effects on nitrifiers and denitrifiers, and mineral N distribution, can modify the efficiency of nitrification inhibitors in reducing N2O emissions from livestock slurry.

Effects of submerged vegetation on sediment nitrogen-cycling bacterial communities in Honghu Lake (China)

Sediment nitrogen (N) cycling is an important biological removal process for N permanently and driven by N-cycling microbial community. There is a growing interest in interactions between submerged vegetation (SV) and sediment N-cycling bacterial community, because of the close link between rooted aquatic plants and the sediment microbes. However, the effects of SV on the sediment N-cycling bacterial community are still controversial. Furthermore, the discrimination of direct and indirect effects of SV on the N-cycling bacterial community remains unclear. Here, we investigated the biomass and species richness of SV and determined the corresponding environment factors (water quality and sediment properties) in Honghu Lake (China). We also used functional genes as markers to unveil the bacterial diversity and community composition and abundance in lake sediments. Our results showed that biomass and species richness of SV affected the composition, diversity and abundance of sediment N-cycling bacterial communities through improving lake water quality and sediment properties. With the increasing richness and abundance of SV, the diversity of most N-cycling bacterial assemblages including nitrifying, denitrifying and DNRA bacteria decreased, while the abundance increased. However, the anammox bacterial assemblage in sediments showed inverse trends. Sediment carbon vs. nitrogen (C:N) ratio negatively affected the abundance of amoA and nirS + nirK + nosZ bacterial assemblages. Additionally, due to the presence of SV, positive interactions among N-cycling bacterial assemblages were found, such as amoA and nrfA bacterial assemblages. Overall, our findings confirmed the significant effects of SV on the N-cycling bacterial community structure and abundance. Moreover, the direct effects of SV on the N-cycling bacterial community and the indirect effects through altering the sediment C were clarified in our study. Our results casted a new light on the negative effects of high C:N ratio. From the study, we made a conclusion that the better SV develops, the greater nitrogen removal occurs in lake sediments.

Fertilization strategies affect soil properties and abundance of N-cycling functional genes in an acidic agricultural soil

The nitrogen (N) cycle process in agricultural soil is affected by fertilization. However, there is limited information on the abundance of N-cycling microbial community and its response to the different fertilization strategies in acidic soil. Using quantitative PCR, we investigated the microbial abundance involving 11 functional genes of the N cycle (nifH, chiA, aprA, archaeal and bacterial amoA, napA, narG, nirS, nirK, norB and nosZ) in an acidic agricultural soil. The soils were subjected to no fertilizer (Control), chemical NPK fertilizer (NPK), NPK combined with quicklime (NPKCa), NPK combined with crop residues (NPKS), NPK combined with quicklime and crop residues (NPKSCa), NPK combined with manure (NPKM) and only manure (M) fertilization for 27 years. Results showed that fertilization improved crop yield and soil nutrient availability, changed soil pH levels and affected the abundance of N-cycling functional genes. Manure application markedly increased the abundance of all genes and was associated with the largest shift in community structure. The variation of microbial community in the fertilization treatments (NPK, NPKCa, NPKS, NPKSCa, M and NPKM) resulted from the distinct contribution of functional genes, in which the nifH, napA and archaeal amoA genes highly contributed to the community variations between manure fertilization and Control. Soil variables, particularly pH, NO3−-N and AP, significantly affected the N-cycling microbial community structure. Overall, the different fertilization strategies demonstrated varying influential patterns on the N-cycling microbial abundance and community by altering soil properties and affecting specific functional gene abundance; in particular, manure treatments had a greater influence compared to chemical fertilizers.