amoA Gene Abundance Research Articles

Chamomile plant material effects on soil nitrogen dynamics and ammonia-oxidizers to mitigate greenhouse gas emissions from maize fields

Commonly, agricultural practices utilize urea as a nitrogen (N) source, which can lead to N losses to the environment. Application of urease inhibitors (UIs) or nitrification inhibitors (NIs) can mitigate these losses to some extent but not completely. A preliminary study showed that 2-Cyclopenten-1-one (CCO), an organic compound derived from chamomile (Chamomila recutita L.) can inhibit soil urease activity and also nitrification and therefore has the potential to act as a dual-function inhibitor for decreasing fertilizer-induced N losses and increasing N use efficiency. In this study, a field experiment was conducted to evaluate the effect of CCO on N transformations and NH3, N2O and CH4 emissions from soil, and monitor changes in the ammonia oxidizer amoA gene abundance. Three replicates of five fertilized treatments were included: Urea (fertilizer only), thiophosphoric triamide (NBPT) (a urease inhibitor), dicyandiamide (DCD) (a widely used synthetic nitrification inhibitor), NBPT combined with DCD, and CCO, along with an unfertilized control.The results indicated that, the ability of CCO to reduce N losses and improve maize yield was potentially superior to DCD, NBPT, or the combination of DCD and NBPT. Compared with the urea treatment, the addition of CCO significantly reduced urease activity and nitrification, which subsequently increased the content of NH4+-N and decreased the content of NO3--N in the soil during the maize growth period. Moreover, the cumulative NH3 and N2O emissions, global warming potential (GWP) and greenhouse gas intensity (GHGI) of the maize field were lowest from the CCO applied plots during the growth season, decreasing by 32.5 %, 21.94 %, 7.69 % and 20.92 %, respectively, compared to urea treatment. Notably, CCO application significantly reduced the abundance of ammonia-oxidizing archaea and bacteria in soil. These results suggest that CCO is effective for suppressing urease activity and the nitrification rate in soil and has the potential to be a novel naturally sourced dual-purpose inhibitor.

Disclosing the ecological implications of heavy metal disturbance on the microbial N-transformation process in the ocean tidal flushing urban estuary

Anthropogenic source of heavy metals in the estuary-coastal sediment could cause serious environmental concerns. Thus, much needs to be concerned about the linkages between bacterial community, microbial nitrogen processes and heavy metals accumulation in the ocean tidal flush urban estuary where the river meet the sea. In this study, Illumina sequencing analysis and quantitative PCR (qPCR) were adopted to detect the effect of heavy metals on the bacterial community and nitrogen transformation genes from vertical sediment profiles in the four tidal flush estuaries. Results indicated that bacterial community and structure varied greatly from the vertical and spatial distribution. The vertical distribution of heavy metals varied greatly in the different groups. It was concluded that the accumulation of heavy metals mainly occurred in the medium layer of the depth from 40 cm to 70 cm. Heavy metals were found to pose a significant influence on the bacterial community and nitrogen transformation genes. The abundance of denitrification genes was highly associated with Cr, Zn, As and the abundance of amoA was vulnerable to the influence of Ni. Both denitrification and nitrification genes were greatly affected by C and N content, which indicated that the enhanced nitrogen transformation activities mainly occurred in the top layer sediment. Our results indicated that the bacterial community and nitrogen transformation functional genes were sensitive to the variation of heavy metals and nutrient content, which will provide new insight into the microbial N-cycling process influenced by pollutant and show a potential strategy for discharge management in the circumstances of estuary-coastal zones.

Microplastics drive nitrification by enriching functional microorganisms in aquaculture pond waters

The plastisphere refers to biofilm formation on the microplastic (MP) surface, but its subsequent functions, especially driving the nitrogen biogeochemical cycle, are rarely studied. Here, MPs were incubated in the pelagic water and benthic water-sediment interface of an aquaculture pond, and the two corresponding microcosms amended with incubated plastisphere were simulated. The results showed decreased ammonia concentrations and increased nitrification rates in microcosms with either pelagic or benthic plastispheres. To uncover the possible mechanisms, the community structure and function of the plastisphere were investigated. As clarified by 16S rRNA, the community diversity of the pelagic plastisphere was significantly higher than that of the corresponding hydrosphere. Plastisphere communities, especially those incubated in pelagic water, were separated from the hydrosphere. Moreover, the abundance of Proteobacteria increased while the abundance of Cyanobacteria decreased in both plastispheres. Metagenome further revealed that the abundance of amoA and annotated Nitrososphaeraceae_archaeon and hao and affiliated Nitrosomonas_europaea, which contributed to ammonia oxidation to nitrite, was higher in the benthic plastisphere. Comparing the pelagic plastisphere with the corresponding hydrosphere, however, the abundance of nxrA and annotated Nitrobacter hamburgensis and nxrB and the affiliated Nitrospira moscoviensis, which are involved in nitrite oxidation, was more abundant in the plastisphere. These findings suggest that the plastisphere might selectively enrich functional microorganisms and genes in a habitat-dependent manner to promote nitrification in aquaculture ponds.

Temporal profiling resolves the drivers of microbial nitrogen cycling variability in coastal sediments

Here we describe the potential for sediment microbial nitrogen-cycling gene (DNA) and activity (RNA) abundances to spatially resolve coastal areas impacted by seasonal variability in external nutrient inputs. Three sites were chosen within a nitrogen-limited embayment, Port Phillip Bay (PPB), Australia that reflect variability in both proximity to external nutrient inputs and the dominant form of available nitrogen. At three sediment depths (0–1; 1–5; 5–10 cm) across a 2 year study key genes involved in nitrification (archaeal amoA and bacterial β-amoA), nitrite reduction (clade I nirS and cluster I nirK, archaeal nirK-a), anaerobic oxidation of ammonium (anammox 16S rRNA phylogenetic marker) and nitrogen fixation (nifH) were quantified. Sediments impacted by a dominance of organic nitrogen inputs were characterised at all time-points and to sediment depths of 10 cm by the highest transcript abundances of archaeal amoA and archaeal nirk-a. Proximity to a dominance of external nitrate inputs was associated with the highest transcript abundances of nirS which temporally co-varied with seasonal changes in sediment nitrate. Sediments isolated from external inputs displayed the greatest depth-specific decrease in quantifiable transcript abundances. In these isolated sediments bacterial β-amoA transcripts were temporally associated with increased sediment ammonium levels. Across this nitrogen limited system variability in the abundance of bacterial β-amoA, archaeal amoA, archaeal nirk-a or nirS transcripts from the sediment surface (0–1 and 5 cm) demonstrated a capacity to improve our ability to monitor coastal zones impacted by anthropogenic nitrogen inputs. Specifically, the spatial detection sensitivity of bacterial β-amoA transcripts could be developed as a metric to determine spatiotemporal impacts of large external loading events. This temporal study demonstrates a capacity for microbial activity metrics to facilitate coastal management strategies through greater spatial resolution of areas impacted by external nutrient inputs.

Snow Exclusion Does Not Affect Soil Ammonia-Oxidizing Bacteria and Archaea Communities

Soil ammonia-oxidizing microorganisms play important roles in nitrogen (N) cycling in cold ecosystems, but how changes in snow cover will affect their distribution and associated functional characteristics remains unclear. A snow manipulation experiment was conducted to explore the effects of snow exclusion on soil ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) communities and functional characteristics in a spruce forest in the eastern Tibet Plateau. Results showed that the amoA gene abundance and community composition of AOA and AOB did not differ between snow regimes but varied among winter periods. AOA and AOB gene abundances showed a decreasing trend during the snow cover melting period. During the deep snow cover period, Thaumarchaeota and Crenarchaeota in the AOA community decreased significantly, while Proteobacteria and Nitrosospira in the AOB community increased significantly. The main factors affecting the changes in AOA and AOB community diversity and composition were soil MBN, nitrate nitrogen, and temperature, while AOA and AOB community diversity and composition were also significantly correlated with soil enzyme activities related to N cycling. These results recommend that the season-driven variations strongly affected soil ammonia-oxidizing community and functional characteristics more than momentary snow cover change. Such findings offer new insights into how soil N-cycling processes would respond to reduced snowfall in high-altitude regions.

Manure combined with biochar reduces rhizosphere nitrification potential and amoA gene abundance of ammonia-oxidizing microorganisms in acid purple soil

Fertilization and soil environmental factors determine the niche differentiation of ammonia-oxidizing microorganisms, thereby affecting the ammonia-oxidation process. However, the characteristics of rhizosphere potential nitrification rate (PNR), ammonia-oxidizing archaea (AOA), and ammonia-oxidizing bacteria (AOB) population under manure combined biochar are unclear. In this study, a pot experiment growing lemon was conducted, setting up six treatments, namely no fertilization (CK), chemical fertilizer (CF), manure (M), chemical fertilizer combined with biochar (CFBC), manure combined with biochar (MBC), fresh manure combined with biochar (FMBC). We investigated the effects of manure, chemical fertilizer and manure combined with biochar on soil physical-chemical properties, PNR, and the amoA gene abundance, diversity and structure of AOA and AOB in the rhizosphere. Our results showed that compared with CK treatment, CF and CFBC increases PNR by 19.3–34.7 % in the rhizosphere, M, MBC and FMBC decrease PNR about 60.0 %; CF and CFBC increased AOA amoA gene copies by 40.2–101.7 %, while M, MBC and FMBC treatments decreased it by 66.5–81.9 %, five fertilization treatments decreased the AOB amoA gene copy by 33.1–81.9 %. The redundancy analysis showed that chemical fertilizer and manure combined with biochar affects the population structure of AOA and AOB by pH, available phosphorus (AP), ratio of carbon to nitrogen (C/N), soil moisture content (SMC), nitrate nitrogen (NO3−-N) and ammonium nitrogen (NH4+-N) in the rhizosphere. The results of partial least squares indicated that pH, total nitrogen (TN), AOA amoA gene copy and AOA Shannon index were main factors affecting nitrification potential in the rhizosphere. Therefore, AOA dominates the ammonia oxidation process in the acid purple soil under the condition of chemical fertilizer and pig manure combined with rice husk biochar. Meanwhile, fertilization affected ammonia oxidation process by regulating soil pH and TN in lemon rhizosphere soil.

Nutrient status of integrated rice-crayfish system impacts the microbial nitrogen-transformation processes in paddy fields and rice yields.

Increasing rice yield is essential for alleviating global food crisis. High soil nutrient level guarantees high rice yields in conventional rice monoculture (RM) systems, but excessive unconsumed nutrients act as pollutants and can even threaten rice growth. The integrated rice-crayfish (IRC) system aims to transfer the excess nutrients from crayfish to paddy fields to improve the comprehensive utilization rate of nutrients and create additional profits, while the responding characteristics of IRC microbial communities in paddy fields and rice yields to the nutrient status remain unclear. Considering the crucial roles of microbiomes in promoting nutrient cycling for crop absorption in rice production progresses, the composition and functional characteristics of soil microbial communities from six IRC farms with variant nutrient statuses in the Yangtze River Delta were surveyed in this study. Compared with RM systems, IRC systems with appropriately improved (p < 0.05) soil quality created favorable nutrient (FN) status accompanied by 15% rice yields increase, while IRC systems with extremely high nutrients (HN) status (p < 0.01) accompanied by 14% rice yields reduction. Soil microbial diversity and network complexity were maintained in FN-IRC systems, but declined in HN-IRC systems, with the Shannon index significantly decreased by 9.2% and network density decreased from 0.135 (in RM) to 0.062. In the FN-IRC systems, the keystone taxa identified by co-occurrence networks displayed inextricably positive correlations with soil nitrification potential (calculated by normalization of amoA gene abundance) and rice yields. While in HN-IRC systems, the large loss of keystone taxa might limit soil nitrogen fixation potential (calculated by normalization of nifH gene abundance), and further rice yields. Our study indicates that soil nutrient management in IRC systems claim attention, and the improvement of nitrogen metabolism is the key to realize agricultural cleaner production.

Nitrogen process in stormwater bioretention: effect of the antecedent dry days on the relative abundance of nitrogen functional genes.

In this study, we evaluated the relative abundance of nitrogen functional genes (amoA, nirK and nirS) involved in ammonia oxidation and denitrification bacteria in laboratory-scale bioretention columns in response to environmental factors (e.g., moisture content, pH, soil organic matter, soil nitrogen) under different antecedent dry days (ADDs). We observed a decrease tendency of the relative abundance of ammonia-oxidizing bacteria at first and then increased when increasing ADDs from 1 to 22 day, while the relative abundance of denitrifying bacteria showed a downward trend. The abundance of bacteria gene amoA was positively associated with soil ammonia nitrogen concentration (r2 = 0.389, p < 0.05) and soil organic matter concentration (r2 = 0.334, p < 0.05), while the abundance of bacteria gene nirS was positively correlated with soil ammonia nitrogen (r2 = 0.730, p < 0.01), soil organic matter (r2 = 0.901, p < 0.01) and soil total nitrogen (r2 = 0.779, p < 0.01). Furthermore, gene counts for bacteria gene nirS were correlated negatively with plant root length (r2 = 0.364, p < 0.05) and plant biomass (r2 = 0.381, p < 0.05). Taken together, these results suggest that both nitrification and denitrification can occur in bioretention systems, which can be affected by environmental factors.

Nitrogen transformation genes and ammonia emission from soil under biochar and urease inhibitor application

Ammonia (NH3) volatilization is the main pathway, by which nitrogen (N) losses from calcareous and alkaline soils occur, and is responsible for economic losses and environmental hazards. We hypothesize that the application of biochar and urease inhibitor (NBPT) can mitigate NH3 production from urea by reducing its hydrolysis and the abundance of related microbial genes. To clarify this hypothesis, soil N fluxes and the genes controlling hydrolysis and nitrification were investigated under the combined application of urea (0, 75, 150, 225 and 300 kg N ha−1), biochar (7.5 Mg ha−1) and NBPT (2 %) for maize under field conditions. Soil moisture, temperature, pH, mineral N, NH3, urease activity (UA) and N cycle related genes (ureC, amoB and amoA) were quantified each 6–17 days during two growing seasons. The combined application of NBPT and biochar reduced NH3 by 13% of the control, which is twofold higher than the separate additions. All investigated parameters were raised with increasing N rate except pH, which was reduced. Biochar and NBPT mediated these effects of N fertilizer by increasing the soil pH, genes abundance, ammonium and nitrate contents, and reducing urea hydrolysis. Biochar maximized the ammonium and nitrate contents one week and three weeks following the N urea application, respectively. Accordingly, NH3 and UA peaked directly after N fertilization; nevertheless, NBPT delayed the peaks for three weeks and reduced their intensity by 24 % and 36 %, respectively, under the addition of 300 kg N ha−1. NBPT reduced ureC and amoB genes abundance by 14 % and 8 %, respectively, and increased amoA by 3 %. Biochar increased amoA and amoB abundance and nitrate content, while reduced the ureC gene. Biochar and/or NBPT reduced the negative effects of the soil pH on ureC gene. NBPT mitigated the reduction effects of pH on the amoB and amoA genes. In conclusion, NBPT mitigated NH3 emission by decreasing the ureC gene and UA, whereas biochar had a synergistic effect with NBPT by increasing amoB and amoA abundance and accelerating nitrification.

A meta-analysis to examine whether nitrification inhibitors work through selectively inhibiting ammonia-oxidizing bacteria.

Nitrification inhibitor (NI) is often claimed to be efficient in mitigating nitrogen (N) losses from agricultural production systems by slowing down nitrification. Increasing evidence suggests that ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) have the genetic potential to produce nitrous oxide (N2O) and perform the first step of nitrification, but their contribution to N2O and nitrification remains unclear. Furthermore, both AOA and AOB are probably targets for NIs, but a quantitative synthesis is lacking to identify the “indicator microbe” as the best predictor of NI efficiency under different environmental conditions. In this present study, a meta-analysis to assess the response characteristics of AOB and AOA to NI application was conducted and the relationship between NI efficiency and the AOA and AOB amoA genes response under different conditions was evaluated. The dataset consisted of 48 papers (214 observations). This study showed that NIs on average reduced 58.1% of N2O emissions and increased 71.4% of soil concentrations, respectively. When 3, 4-dimethylpyrazole phosphate (DMPP) was applied with both organic and inorganic fertilizers in alkaline medium soils, it had higher efficacy of decreasing N2O emissions than in acidic soils. The abundance of AOB amoA genes was dramatically reduced by about 50% with NI application in most soil types. Decrease in N2O emissions with NI addition was significantly correlated with AOB changes (R2 = 0.135, n = 110, P < 0.01) rather than changes in AOA, and there was an obvious correlation between the changes in concentration and AOB amoA gene abundance after NI application (R2 = 0.037, n = 136, P = 0.014). The results indicated the principal role of AOB in nitrification, furthermore, AOB would be the best predictor of NI efficiency.

Machine Learning Prediction of Nitrification From Ammonia- and Nitrite-Oxidizer Community Structure.

Accurately modeling nitrification and understanding the role specific ammonia- or nitrite-oxidizing taxa play in it are of great interest and importance to microbial ecologists. In this study, we applied machine learning to 16S rRNA sequence and nitrification potential data from an experiment examining interactions between cropping systems and rhizosphere on microbial community assembly and nitrogen cycling processes. Given the high dimensionality of microbiome datasets, we only included nitrifers since only a few taxa are capable of ammonia and nitrite oxidation. We compared the performance of linear and nonlinear algorithms with and without qPCR measures of bacterial and archaea ammonia monooxygenase subunit A (amoA) gene abundance. Our feature selection process facilitated the identification of taxons that are most predictive of nitrification and to compare habitats. We found that Nitrosomonas and Nitrospirae were more frequently identified as important predictors of nitrification in conventional systems, whereas Thaumarchaeota were more important predictors in diversified systems. Our results suggest that model performance was not substantively improved by incorporating additional time-consuming and expensive qPCR data on amoA gene abundance. We also identified several clades of nitrifiers important for nitrification in different cropping systems, though we were unable to detect system- or rhizosphere-specific patterns in OTU-level biomarkers for nitrification. Finally, our results highlight the inherent risk of combining data from disparate habitats with the goal of increasing sample size to avoid overfitting models. This study represents a step toward developing machine learning approaches for microbiome research to identify nitrifier ecotypes that may be important for distinguishing ecotypes with defining roles in different habitats.

Application of Reeds as Carbon Source for Enhancing Denitrification of Low C/N Micro-Polluted Water in Vertical-Flow Constructed Wetland

Constructed wetlands have been applied to micro-polluted rivers and lakes. However, they often show poor nitrogen removal efficiency due to insufficient carbon sources for complete denitrification in the waters. In this study, a vertical-flow wetland system was built, in which reeds as a carbon source were added in the middle layer of the substrate. Thereby, the effect of the reed carbon source on denitrification of micro-polluted rivers and lakes with a low C/N ratio in the wetland and the denitrification mechanism were studied. The results showed that the concentrations of NH4+-N, NO3−-N and NO2−-N in the effluent of the constructed wetland were reduced to 0.17–0.35, 0.20–0.49 and 0.01–0.02 mg/L after adding the reed carbon source, and the removal efficiencies of the system for NH4+-N and NO3−-N reached 93.84% and 84.69%, respectively. The abundances of nirK, nirS, hzo and nrfA genes in the wetland substrate increased by 95.51%, 54.96%, 52.89% and 731.95%, respectively, which was considered to be related to the enhanced denitrification, anammox and dissimilatory nitrate reduction to ammonium of the wetland system. Reed planting promoted the increased abundances of amoA and nxrB genes, which might play a positive role in enhancing nitrification in wetland systems. The result of this study may provide a theoretical basis for the ecological restoration of low C/N micro-polluted water bodies.

Non-target impacts of pesticides on soil N transformations, abundances of nitrifying and denitrifying genes, and nitrous oxide emissions

Agriculture is the leading contributor to global nitrous oxide (N2O) emissions, mostly from soils. We examined the non-target impacts of four pesticides on N transformations, N cycling genes and N2O emissions from sugarcane-cropped soil. The pesticides, including a herbicide glyphosate (GLY), an insecticide imidacloprid (IMI), a fungicide methoxy ethyl mercuric chloride (MEMC) and a fumigant methyl isothiocyanate (MITC), were added to the soil and incubated in laboratory at 25 °C. The soil microcosms were maintained at two water contents, 55 % and 90 % water holding capacity (WHC), to simulate aerobic and partly anaerobic conditions, respectively. Half of the soil samples received an initial application of KNO3 and were then maintained at 90 % WHC for 38 d, whilst the other half received (NH4)2SO4 and were maintained at 55 % WHC for 28 d followed by 10 d at 90 % WHC to favour denitrification. Responses of individual functional genes involved in nitrification and denitrification to the pesticides and their relationships to N2O emissions varied with time and soil water. Overall, MITC had pronounced repressive effects on AOA and AOB amoA gene abundances and gross nitrification. Under 55 % WHC during the initial 28 d, N2O emissions were low for all treatments (≤62 μg N kg−1 soil). However, under 90 % WHC (either during the first 28 d or the increase in water content from 55 to 90 % WHC after 28 d) the cumulative N2O emissions increased markedly. Overall, under 90 % WHC the cumulative N2O emissions were 19 (control) to 79-fold (MITC) higher than under 55% WHC; with the highest emissions observed in the MITC treatment (3140 μg N kg−1 soil). This was associated with increases in gross nitrate consumption rates and abundances of denitrifying genes (nirK, nirS and qnorB). Therefore, to minimise N2O emissions, MITC should not be applied to field under wet conditions favouring denitrification.

Nitrogen deposition stimulated winter nitrous oxide emissions from bare sand more than biological soil crusts in cold desert ecosystem

Dryland ecosystems are often nitrogen-limited, and small nitrogen inputs may produce large responses to dryland ecological processes, such as gaseous nitrogen emission. The effect of increased anthropogenic nitrogen deposition on N2O and NO emissions in desert ecosystems is unclear, especially in non-growing seasons when the surface is covered with snow. In this study, nitrogen applications were performed on biological soil crusts (lichen crust and moss crust, bare sand for control) in the Gurbantunggut Desert, Northwest China. We measured the fluxes of N2O and NO and related nitrogen cycle functional gene abundances in winter for three-years period. Nitrogen addition significantly affected N2O emissions and increased the abundances of key functional gene for nitrogen cycle, while it only slightly influenced NO emissions. These effects of nitrogen addition depended on composition of biological soil crusts. For bare sand and lichen crust, nitrogen addition significantly increased N2O emissions, whereas for moss crust, only a negligible effect was observed. Meanwhile, significant differences in nitrogen cycle functional gene abundances were found among different composition of biological soil crusts. Abundance of amoA, narG, and nosZ genes were highly related to N2O and NO emissions. Thus, our results indicate that gaseous nitrogen emissions were generally increased by nitrogen addition through their effects on related functional microbial groups. The effects were regulated by composition of biological soil crusts which can buffer the effects of increasing nitrogen addition during winter.

Improving Nitrogen Use Efficiency in Aerobic Rice Based on Insights Into the Ecophysiology of Archaeal and Bacterial Ammonia Oxidizers.

The abundance and structural composition of nitrogen (N) transformation-related microbial communities under certain environmental conditions provide sufficient information about N cycle under different soil conditions. This study aims to explore the major challenge of low N use efficiency (NUE) and N dynamics in aerobic rice systems and reveal the agronomic-adjustive measures to increase NUE through insights into the ecophysiology of ammonia oxidizers. Water-saving practices, like alternate wetting and drying (AWD), dry direct seeded rice (DDSR), wet direct seeding, and saturated soil culture (SSC), have been evaluated in lowland rice; however, only few studies have been conducted on N dynamics in aerobic rice systems. Biological ammonia oxidation is majorly conducted by two types of microorganisms, ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). This review focuses on how diversified are ammonia oxidizers (AOA and AOB), whose factors affect their activities and abundance under different soil conditions. It summarizes findings on pathways of N cycle, rationalize recent research on ammonia oxidizers in N-cycle, and thereby suggests adjustive agronomic measures to reduce N losses. This review also suggests that variations in soil properties significantly impact the structural composition and abundance of ammonia oxidizers. Nitrification inhibitors (NIs) especially nitrapyrin, reduce the nitrification rate and inhibit the abundance of bacterial amoA without impacting archaeal amoA. In contrast, some NIs confine the hydrolysis of synthetic N and, therefore, keep low NH4+-N concentrations that exhibit no or very slight impact on ammonia oxidizers. Variations in soil properties are more influential in the community structure and abundance of ammonia oxidizers than application of synthetic N fertilizers and NIs. Biological nitrification inhibitors (BNIs) are natural bioactive compounds released from roots of certain plant species, such as sorghum, and could be commercialized to suppress the capacity of nitrifying soil microbes. Mixed application of synthetic and organic N fertilizers enhances NUE and plant N-uptake by reducing ammonia N losses. High salt concentration promotes community abundance while limiting the diversity of AOB and vice versa for AOA, whereas AOA have lower rate for potential nitrification than AOB, and denitrification accounts for higher N2 production. Archaeal abundance, diversity, and structural composition change along an elevation gradient and mainly depend on various soil factors, such as soil saturation, availability of NH4+, and organic matter contents. Microbial abundance and structural analyses revealed that the structural composition of AOA was not highly responsive to changes in soil conditions or N amendment. Further studies are suggested to cultivate AOA and AOB in controlled-environment experiments to understand the mechanisms of AOA and AOB under different conditions. Together, this evaluation will better facilitate the projections and interpretations of ammonia oxidizer community structural composition with provision of a strong basis to establish robust testable hypotheses on the competitiveness between AOB and AOA. Moreover, after this evaluation, managing soils agronomically for potential utilization of metabolic functions of ammonia oxidizers would be easier.

Uptake of individual and mixed per- and polyfluoroalkyl substances (PFAS) by soybean and their effects on functional genes related to nitrification, denitrification, and nitrogen fixation

In this study, we set up a soil-microbe-soybean system spiked with PFOA, PFOS, or a PFAS mixture of eight PFAS and investigated the distribution of PFAS in the system and impacts on the abundance and expression level of genes involved in the nitrogen (N) cycle. When soybean was exposed to the PFAS mixtures, synergistic uptake by shoots was detected. PFAS exhibited remarkable impacts on abundance of nitrification and denitrification genes in both bulk soil and rhizosphere as well as expression of N fixation gene in soybean nodules. The abundance of nitrification genes AOA and AOB amoA and denitrification gene nirK was significantly reduced (p < 0.05) in almost all treatments in bulk soil, except PFOA at 10 μg/kg. The abundance of other functional genes, such as nirS and norZ was affected differently depending on PFAS concentrations and sample location, either bulk soil or the rhizosphere. Interestingly, the N fixation gene nifH in soybean nodules was overexpressed by a PFAS mixture at 100 μg/kg. Hence, this work provided in-depth knowledge regarding the distribution of PFAS and their impacts on the N cycle for the studied system. Results from this study provide insights on assessing risks posed by individual or mixed PFAS to soybean.

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.

Impact of anaerobic digestion on reactive nitrogen gas emissions from dairy slurry storage

Biogas digesters are commonly used to treat animal manure/slurry, and abundant digested slurry is generated during the digestion process. Gas emissions from digested and raw slurry may vary with the change in slurry parameters after digestion, but the mechanism is not well understood. Gas emissions from raw dairy slurry (RS) and digested dairy slurry (BS) during 98 days of storage were investigated in this study to evaluate the effects of anaerobic digestion on reactive nitrogen emissions from slurry storage. Results showed that much higher N2O and NO emission and lower NH3 emission was achieved in BS than in RS. The mean gaseous emission of RS and BS accounted for 27.8% ± 6.9% and 17.1% ± 2.3% of the initial TN for NH3, 0.1% ± 0.1% and 3.5% ± 1.6% of the initial TN for N2O, and 0.0% ± 0.0% and 0.2% ± 0.0% of the initial TN for NO, respectively. Among all detected N2O-forming and reducing microbial genes, the abundance of amoA genes was the most closely related to N2O flux (r = 0.54, p < 0.01). More aerobic conditions occurred in BS, and dissolved oxygen (DO) increased to 0.4–1.6 mg L−1 after 35 days because the low organic matter of BS resulted in good infiltration of surface air into the slurry. The increased DO stimulated the growth of Nitrosomonas and the increase in amoA gene copies and contributed to the high N2O and NO emissions in BS through the nitrification process. Vulcanibacillus, Thauera, Castellaniella, and Thermomonas were the major denitrifying bacteria that occurred in BS and caused an incomplete denitrification process, which could be another reason for the increase in N2O and NO emissions from BS. Our study indicated that anaerobic digestion reduced the organic matter content of the slurry and caused an active microbial environment that facilitated the transformation of slurry N to N2O in BS storage, thus lowering the NH3 emission compared with RS storage. Therefore, aside from NH3, N2O should also be preferentially mitigated during BS storage because N2O is a greenhouse gas with high global warming potential.

Soil Microbial Legacy Overrides the Responses of a Dominant Grass and Nitrogen-Cycling Functional Microbes in Grassland Soil to Nitrogen Addition.

Both atmospheric nitrogen (N) deposition and soil microbial legacy (SML) can affect plant performance, the activity of soil N-cycling functional microbes and the relative abundance of N-cycling functional genes (NCFGs). In the grassland vegetation successional process, how the interaction of SML and N deposition affects the performance of dominant grass and NCFGs remains unclear. Therefore, we planted Leymus chinensis, a dominant grass in the Songnen grassland, in the soil taken from the early, medium, late, and stable successional stages. We subjected the plants to soil sterilization and N addition treatments and measured the plant traits and NCFG abundances (i.e., nifH, AOB amoA, nirS, and nirK). Our results showed the biomass and ramet number of L. chinensis in sterilized soil were significantly higher than those in non-sterilized soil, indicating that SML negatively affects the growth of L. chinensis. However, N addition increased the plant biomass and the AOB amoA gene abundance only in sterilized soils, implying that SML overrode the N addition effects because SML buffered the effects of increasing soil N availability on NCFGs. Therefore, we emphasize the potential role of SML in assessing the effects of N deposition on dominant plant performance and NCFGs in the grassland vegetation succession.

Ammonia oxidizing bacteria dominate soil nitrification under different fertilization regimes in black soils of northeast China

Although autotrophic ammonia oxidizing archaea (AOA) and bacteria (AOB) and comammox coexist in soils, their abundances and contributions to nitrification are not well addressed, especially in arable black soils. In this study, using the nitrification inhibitors acetylene and octyne, the contributions to nitrification potential provided by different ammonia oxidizers and heterotrophic nitrifiers in four long-term fertilization treatments, i.e., no fertilization (NoF), manure fertilization (M), chemical fertilization (CF), and chemical fertilization plus manure (CFM), were investigated in samples collected from the northern, middle and southern black soil regions of northeast China. The amoA gene abundances of AOA, AOB, comammox Clade A and comammox Clade B in these soils were quantified by qPCR. The results showed that the amoA gene abundances of the three ammonia oxidizers tended to increase from the northern to southern sites, while the opposite trend was observed in the ratios of AOA/AOB and comammox Clade B/Clade A; the different fertilization treatments significantly influenced the amoA abundances, and the effects varied with the ammonia oxidizers and across the sites. Autotrophic ammonia oxidizers accounted for 62%–100% of the gross soil nitrification potential across all soils. Among them, AOB dominated soil autotrophic nitrification in the fertilized soils, while no fertilization may have increased the nitrification potential provided by AOA plus comammox. Pairwise analysis showed that the amoA gene abundances of AOA and AOB were significantly and positively correlated with their respective nitrification-driven potentials. Correlation heatmaps indicated that soil pH and inorganic nitrogen content were the main soil factors related to nitrification activities and the abundances of the ammonia oxidizers. Our findings highlight that AOB are the dominant ammonia oxidizers contributing to nitrification in arable fertilized black soils, although AOA and comammox are highly abundant.