Ammonia-oxidizing Archaea Populations Research Articles

Dynamic Responses of Ammonia-Oxidizing Archaea and Bacteria Populations to Organic Material Amendments Affect Soil Nitrification and Nitrogen Use Efficiency.

Organic material amendments have been proposed as an effective strategy to promote soil health by enhancing soil fertility and promoting nitrogen (N) cycling and N use efficiency (NUE). Thus, it is important to investigate the extent to which the structure and function of ammonia-oxidizing archaea (AOA) and bacteria (AOB) differentially respond to the organic material amendments in field settings. Here, we conducted a 9-year field experiment to track the responses of AOA and AOB populations to the organic material amendments and measured the potential nitrification activity (PNA), plant productivity, and NUE in the plant rhizosphere interface. Our results revealed that the organic material amendments significantly enhanced the abundance and diversity of AOA and AOB populations. Further, significant differences were observed in the composition and co-occurrence network of AOA and AOB. A higher occurrence of potential competitive interactions between taxa and enumerated potential keystone taxa was observed in the AOA-AOB network. Moreover, we found that AOA was more important than AOB for PNA under the organic material amendments. Structural equation modeling suggested that the diversity of AOA and AOB populations induced by the potential competitive interactions with keystone taxa dynamically accelerated the rate of PNA, and positively affected plant productivity and NUE under the organic material amendments. Collectively, our study offers new insights into the ecology and functioning of ammonia oxidizers and highlights the positive effects of organic material amendments on nitrogen cycling dynamics.

From rags to enriched: metagenomic insights into ammonia-oxidizing archaea following ammonia enrichment of a denuded oligotrophic soil ecosystem.

SummaryStored topsoil acts as a microbial inoculant for ecological restoration of land after disturbance, but the altered circumstances frequently create unfavourable conditions for microbial survival. Nitrogen cycling is a critical indicator for ecological success and this study aimed to investigate the cornerstone taxa driving the process. Previous in silico studies investigating stored topsoil discovered persistent archaeal taxa with the potential for re‐establishing ecological activity. Ammonia oxidization is the limiting step in nitrification and as such, ammonia‐oxidizing archaea (AOA) can be considered one of the gatekeepers for the re‐establishment of the nitrogen cycle in disturbed soils. Semi‐arid soil samples were enriched with ammonium sulfate to promote the selective enrichment of ammonia oxidizers for targeted genomic recovery, and to investigate the microbial response of the microcosm to nitrogen input. Ammonia addition produced an increase in AOA population, particularly within the genus Candidatus Nitrosotalea, from which metagenome‐assembled genomes (MAGs) were successfully recovered. The Ca. Nitrosotalea archaeon candidates' ability to survive in extreme conditions and rapidly respond to ammonia input makes it a potential bioprospecting target for application in ecological restoration of semi‐arid soils and the recovered MAGs provide a metabolic blueprint for developing potential strategies towards isolation of these acclimated candidates.

Diverse ecophysiological adaptations of subsurface Thaumarchaeota in floodplain sediments revealed through genome-resolved metagenomics.

The terrestrial subsurface microbiome contains vastly underexplored phylogenetic diversity and metabolic novelty, with critical implications for global biogeochemical cycling. Among the key microbial inhabitants of subsurface soils and sediments are Thaumarchaeota, an archaeal phylum that encompasses ammonia-oxidizing archaea (AOA) as well as non-ammonia-oxidizing basal lineages. Thaumarchaeal ecology in terrestrial systems has been extensively characterized, particularly in the case of AOA. However, there is little knowledge on the diversity and ecophysiology of Thaumarchaeota in deeper soils, as most lineages, particularly basal groups, remain uncultivated and underexplored. Here we use genome-resolved metagenomics to examine the phylogenetic and metabolic diversity of Thaumarchaeota along a 234 cm depth profile of hydrologically variable riparian floodplain sediments in the Wind River Basin near Riverton, Wyoming. Phylogenomic analysis of the metagenome-assembled genomes (MAGs) indicates a shift in AOA population structure from the dominance of the terrestrial Nitrososphaerales lineage in the well-drained top ~100 cm of the profile to the typically marine Nitrosopumilales in deeper, moister, more energy-limited sediment layers. We also describe two deeply rooting non-AOA MAGs with numerous unexpected metabolic features, including the reductive acetyl-CoA (Wood-Ljungdahl) pathway, tetrathionate respiration, a form III RuBisCO, and the potential for extracellular electron transfer. These MAGs also harbor tungsten-containing aldehyde:ferredoxin oxidoreductase, group 4f [NiFe]-hydrogenases and a canonical heme catalase, typically not found in Thaumarchaeota. Our results suggest that hydrological variables, particularly proximity to the water table, impart a strong control on the ecophysiology of Thaumarchaeota in alluvial sediments.

Ammonia-oxidizing archaea have similar power requirements in diverse marine oxic sediments

Energy/power availability is regarded as one of the ultimate controlling factors of microbial abundance in the deep biosphere, where fewer cells are found in habitats of lower energy availability. A critical assumption driving the proportional relationship between total cell abundance and power availability is that the cell-specific power requirement keeps constant or varies over smaller ranges than other variables, which has yet to be validated. Here we present a quantitative framework to determine the cell-specific power requirement of the omnipresent ammonia-oxidizing archaea (AOA) in eight sediment cores with 3–4 orders of magnitude variations of organic matter flux and oxygen penetration depth. Our results show that despite the six orders of magnitude variations in the rates and power supply of nitrification and AOA abundances across these eight cores, the cell-specific power requirement of AOA from different cores and depths overlaps within the narrow range of 10−19–10−17 W cell−1, where the lower end may represent the basal power requirement of microorganisms persisting in subseafloor sediments. In individual cores, AOA also exhibit similar cell-specific power requirements, regardless of the AOA population size or sediment depth/age. Such quantitative insights establish a relationship between the power supply and the total abundance of AOA, and therefore lay a foundation for a first-order estimate of the standing stock of AOA in global marine oxic sediments.

Variation, distribution, and diversity of canonical ammonia-oxidizing microorganisms and complete-nitrifying bacteria in highly contaminated ecological restoration regions in the Siding mine area

Canonical ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB) and complete-nitrifying bacteria (comammox) exist in a variety of ecosystems. However, little is known about AOA, AOB and comammox or their contributions to nitrification in the soils of heavily degraded and acidic mine regions. In the present study, the activity, richness, diversity and distribution patterns of AOA, AOB and comammox in the Siding mine area were investigated. Nemerow's multifactor pollution index (PN) values indicated that the soil in all three areas in the Siding mine area was highly contaminated by Cd, Pb, Zn, Mn and Cu. The AOA, AOB and comammox amoA gene copy numbers exhibited significant positive correlations with Pb and Zn levels and PN values, which indicated that the populations of AOA, AOB and comammox underwent adaptation and reproduction in response to pollution from multiple metals in the Siding mine area. Among them, the abundance of AOA was the highest, and AOA may survive better than AOB and comammox under such severely pollution-stressed and ammonia-limited conditions. The phyla Thaumarchaeota and Crenarchaeota may play vital roles in the soil ammonia oxidation process. Unlike AOA, AOB may use soil available phosphorus to help them compete for NH3 and other limiting nutrients with AOA and heterotrophs. Moreover, soil organic matter was the main factor influencing the species diversity of AOB, the β-diversity of AOB and comammox, and the community composition of AOA, AOB and comammox. Our research will help to explain the role and importance of AOA, AOB and comammox in the different ecological restoration regions in the Siding mine area.

Seasonal Prevalence of Ammonia-Oxidizing Archaea in a Full-Scale Municipal Wastewater Treatment Plant Treating Saline Wastewater Revealed by a 6-Year Time-Series Analysis.

Although several molecular-based studies have demonstrated the involvement of ammonia-oxidizing archaea (AOA) in ammonia oxidation in wastewater treatment plants (WWTPs), factors affecting the persistence and growth of AOA in these engineered systems have not been resolved. Here, we show a seasonal prevalence of AOA in a full-scale WWTP (Shatin, Hong Kong SAR) over a 6-year period of observation, even outnumbering ammonia-oxidizing bacteria in the seasonal peaks in 3 years, which may be due to the high bioavailable copper concentrations. Comparative analysis of three metagenome-assembled genomes of group I.1a AOA obtained from the activated sludge and 16S rRNA gene sequences recovered from marine sediments suggested that the seawater used for toilet flushing was the primary source of the WWTP AOA. A rare AOA population in the estuarine source water became transiently abundant in the WWTP with a metagenome-based relative abundance of up to 1.3% over three seasons of observation. Correlation-based network analysis revealed a robust co-occurrence relationship between these AOA and organisms potentially active in nitrite oxidation. Moreover, a strong correlation between the dominant AOA and an abundant proteobacterial organism suggested that capacity for extracellular polymeric substance production by the proteobacterium could provide a niche for AOA within bioaggregates. Together, the study highlights the importance of long-term observation in identifying biotic and abiotic factors governing population dynamics in open systems such as full-scale WWTPs.

The Contribution of Root Turnover on Biological Nitrification Inhibition and Its Impact on the Ammonia-Oxidizing Archaea under Brachiaria Cultivations

Aims: Biological nitrification inhibition (BNI) has been reported as an emerging technology to control soil nitrifier activity for effective N-utilization in cropping systems. Brachiaria have been reported to suppress nitrifier populations by releasing nitrification inhibitors from roots through exudation. Substantial BNI activity has been reported to be present in the root tissues of Brachiaria grasses; however, BNI contribution, such as root turnover, has not been addressed in previous studies. The present study aimed to clarify the contribution of root turnover on BNI under Brachiaria cultivations and its impact on nitrifier populations. Methods: We monitored root growth, changes in ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) numbers, nitrification rate, and available nitrogen (N) content under seven germplasm lines of Brachiaria, for 18 months with seasonal profile sampling. Results: Brachiaria cultivation increased soil NH4+-N, available N, and total soil carbon levels. Though we did not find any correlation between the changes in AOB populations and potential nitrification, the potential nitrification rate decreased when AOA populations decreased. Multiple regression analysis indicated that BNI substances from root tissue turnover had a significant contribution to the BNI function in the field. Conclusion Results indicated that the inhibitory effect of BNI was mostly evident in AOA, and not in AOB, in this study. Brachiaria cvs. ‘Marandu’, ‘Mulato’, and ‘Tupy’ had the most substantial BNI effect among the seven cultivars evaluated. The estimated total BNI activities and available N content of root tissue explained the observed nitrification inhibition. In conclusion, the release of BNI substances through plant decomposition contributes to the decrease in the abundance of AOA, and thus the inhibition of nitrification under Brachiaria cultivation.

Sorgoleone release from sorghum roots shapes the composition of nitrifying populations, total bacteria, and archaea and determines the level of nitrification

Sorgoleone is a secondary sorghum metabolite released from roots. It has allelopathic properties and is considered to inhibit ammonia-oxidizing archaea (AOA) and bacteria (AOB) responsible for the rate-limiting step (ammonia oxidation) in nitrification. Low activity of these microorganisms in soil may contribute to slow down nitrification and reduce nitrogen loss via denitrification and NO3− leaching. The potential nitrification rate (PNR) and the composition of microbial communities were monitored in rhizosphere soil to investigate the growth effect sorghum on biological nitrification inhibition (BNI). A greenhouse pipe experiment was conducted using sorghum lines IS20205 (high-sorgoleone release ability), IS32234 (medium-sorgoleone release ability), 296B (low-sorgoleone release ability), and a control (no plants) combined with fertilization application of 0 or 120 kg N ha−1. We applied nitrogen as ammonium sulfate at 16 days (20 N), 37 days (40 N), and 54 days (60 N). We collected soil solutions at 7.5 cm depths every 3 days and measured the pH and nitrate levels. At 1 and 2.3 months, we sampled the bulk and rhizosphere soils and roots in the 0–10 cm, 10–30 cm, and 30–80 cm depths to determine NO2, mineral N, total N, total C, sorgoleone, the composition of AOA, AOB, and total bacteria and archaea. Sorgoleone was continuously released throughout the 2.3 months’ growth and was significantly higher in IS20205, followed by IS32234 then 296B, which showed shallow levels. The IS2020 5rhizosphere showed lower NO2 and nitrate levels and significant inhibition of AOA populations. However, we did not find significant differences in the abundance of AOB between plant treatments. Multivariate analysis and Spearman’s correlations revealed that sorgoleone as well as environmental factors such as soil pH, soil moisture, NO3−-N, and NH4+-N shape the composition of microbial communities. This study demonstrated that the release of higher amounts of sorgoleone has great potential to inhibit the abundance of AOA and soil nitrification. The breeding of sorghum lines with the ability to release higher amounts of sorgoleone could be a strategic way to improve the biological nitrification inhibition during cultivation.

The dominance of non-halophilic archaea in autotrophic ammonia oxidation of activated sludge under salt stress: A DNA-based stable isotope probing study

Dynamics of nitrification activity, ammonia-oxidizing archaea (AOA) and bacteria (AOB) abundance and active ammonia oxidizers of activated sludge were explored under different salinities. Results showed that specific ammonium oxidation rates were significantly negative with increasing salinity. The responses of AOA and AOB populations to salt stress were distinct. AOA abundance decreased at moderate salinities (2.5, 5 and 7 g L−1) and increased at high salinities (10, 15, 20 and 30 g L−1), while AOB abundance showed opposite tendency. DNA-based stable isotope probing assays indicated AOA exclusively dominated active ammonia oxidation of test samples under different salinities. The active AOA communities retrieved were all non-halophilic and regulated by salinities. Candidatus Nitrosocosmicus exaquare and Ca. Nitrosocosmicus franklandus were the predominantly active AOA in both salt-free and salt-containing microcosms, while 13C-labeled Nitrososphaera viennensis and Ca. Nitrososphaera gargensis were only retrieved from the microcosms amended with 0 and 30 g L−1 salinity, respectively.

Functional relationship between ammonia-oxidizing bacteria and ammonia-oxidizing archaea populations in the secondary treatment system of a full-scale municipal wastewater treatment plant

The abundance of ammonia-oxidizing bacteria and archaea and their amoA genes from the aerobic activated sludge tanks, recycled sludge and anaerobic digesters of a full-scale wastewater treatment plant (WWTP) was determined. Polymerase chain reaction and denaturing gradient gel electrophoresis were used to generate diversity profiles, which showed that each population had a consistent profile although the abundance of individual members varied. In the aerobic tanks, the ammonia-oxidizing bacterial (AOB) population was more than 350 times more abundant than the ammonia-oxidizing archaeal (AOA) population, however in the digesters, the AOA population was more than 10 times more abundant. Measuring the activity of the amoA gene expression of the two populations using RT-PCR also showed that the AOA amoA gene was more active in the digesters than in the activated sludge tanks. Using batch reactors and ddPCR, amoA activity could be measured and it was found that when the AOB amoA activity was inhibited in the anoxic reactors, the expression of the AOA amoA gene increased fourfold. This suggests that these two populations may have a cooperative relationship for the oxidation of ammonia.

Responses of nitrification and denitrification to nitrogen and phosphorus fertilization: does the intrinsic soil fertility matter?

Nitrogen (N) is not only a major regulator of productivity in terrestrial systems but can also be a pollutant. While the effects of fertilizer addition to soil N cycling processes and the associated soil microbial groups have been extensively studied, little attention has been paid to the interaction of fertilizer amendment and the intrinsic fertility of the soil. We conducted a filed study on the Chinese Loess Plateau with soils of two fertility levels (low fertility and high fertility), amended with or without synthetic fertilizer (N and phosphorus, NP). Soil samples were collected three times in a continuous winter wheat cropping system and analyzed for soil potential nitrification (PNA) and denitrification (DEA) rates, nitrifiers and denitrifiers. Populations of ammonia-oxidizing archaea (AOA) and bacteria (AOB) and the resulting PNA were increased by the NP application regardless of intrinsic soil fertility. The NP application increased DEA by 15–228% in the low fertility soil, but decreased DEA by 18–46% in the high fertility soil. Soil denitrifiers also showed divergent responses to NP application in low and high fertility soils, i.e., the abundance of nirS-nitrite reducers increased in the low fertility soil, while nirS- and nirK-nitrite reducers were unchanged in the high fertility soil. The narG-nitrate reducers, AOB and AOA were more responsive to the NP application in the tillering stage (BBCH 25–26) than in other periods, whereas PNA, DEA and denitrifier gene abundance (nirS and nirK) varied more in the flowering stage (BBCH 65). Our results suggest that the responses of soil nitrification/nitrifiers and denitrification /denitrifiers to NP application varied with intrinsic soil fertility levels. The contrasting responses of nitrification and denitrification to NP application in high fertility soil indicate a decoupling of both processes.

Spatial and temporal responses of ammonia-oxidizing bacteria and archaea to organic amendments in rice-wheat rotation system

To determine the effects of organic amendment on communities of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) in rice-wheat rotation ecosystem, a long-term plot experiment with 5 treatments was conducted, and the temporal dynamics (10 growth stages) and spatial distribution (0–100 cm) of AOB and AOA abundance were determined using qPCR. The five treatments were as follows: no fertilizer control (CK), regular fertilization (RT), straw returning (SR), straw biochar returning (SB) and organic-chemical mixed fertilization (MF). The results showed the following. 1) Organic addition did not affect the temporal dynamics of AOB abundance but altered those of AOA abundance. Relative to RT, SR and MF significantly lowered AOB population sizes across the 10 sampling time points by 34.49% and 16.42%, respectively, whereas no significant differences in total AOA population size were observed among the 4 fertilization regimes. 2) Compared with the other treatments, treatments involving organic addition had higher total AOB abundance in the 0–100 cm soil layer. However, SB was the only treatment with organic addition that had a significantly higher total AOA abundance than RT (by 24.91%). 3) Under organic addition, the potential nitrification rate (PNR) was still highly correlated with AOB abundance (P < 0.01) but not AOA abundance.

Seasonal variation and ex-situ nitrification activity of ammonia oxidizing archaea in biofilm based wastewater treatment processes

The abundance of ammonia oxidizing archaea (AOA) and ammonia oxidizing bacteria (AOB) was investigated in full-scale two-stage trickling filters (TF) and moving bed bioreactor (MBBR) treating municipal wastewater. Biofilm samples were collected for 17months from nitrifying TF (NTF), biochemical oxygen demand TF (BTF), and MBBR media. The abundance of AOA and AOB was determined using a quantitative PCR approach targeting the ammonia monooxygenase subunit A gene of archaea and bacteria. AOA were dominant in the NTF and MBBR, while AOB dominated in the BTF. AOA and AOB were more abundant during warmer months, and AOA were detected in the BTF only during warmer months. In laboratory nitrification activity experiments, ammonia oxidation to nitrite decreased when AOA populations from the NTF and MBBR were inhibited, demonstrating that AOA contributed to nitrification. This study has shown that AOA outnumber AOB and contribute to ammonia oxidation in full-scale nitrifying biofilm processes.

Effects of hexaconazole application on soil microbes community and nitrogen transformations in paddy soils

The ecological risks of widely used hexaconazole on soil microbes remain obscure. Thus, a 3-month-long experiment using two typical paddy soils in China (red soil and black soil) was conducted to assess the effects of hexaconazole (0.6 (T1) and 6 (T10) mgkg−1 soil) on the overall microbial biomass, respiratory activity, bacterial abundance and community structure, and nitrogen transformations. Soil was sampled after 7, 15, 30, 60, and 90days of incubation. The half-lives of the two doses of hexaconazole varied from 122 to 135d in the black soil and from 270 to 845d in the red soil. Both dosages of hexaconazole did not affect NH+4-N content, N2-fixing bacterial populations, total bacterial diversity, and community structure, but transitorily decreased the populations of total bacteria in both soil types. In the black soil, T10 negatively affected microbial biomass carbon (MBC) and soil basal respiration (RB), but transitorily increased NO−3–N concentration and ammonia-oxidizing bacteria populations, while T1 had almost no effect on most of the indicators. As for red soil, both concentrations of fungicide significantly, but transitorily, inhibited MBC and RB, while only T10 had a relatively long stimulatory effect on NO−3–N concentration and ammonia-oxidizing archaea populations. This study showed that over application of hexaconazole is indeed harmful to soil microorganisms and may reduce soil quality and increase the risk of nitrogen loss in paddy soils.

Ammonia oxidizer populations vary with nitrogen cycling across a tropical montane mean annual temperature gradient.

Functional gene approaches have been used to better understand the roles of microbes in driving forest soil nitrogen (N) cycling rates and bioavailability. Ammonia oxidation is a rate limiting step in nitrification, and is a key area for understanding environmental constraints on N availability in forests. We studied how increasing temperature affects the role of ammonia oxidizing archaea (AOA) and bacteria (AOB) in soil N cycling and availability by using a highly constrained natural mean annual temperature (MAT) elevation gradient in a tropical montane wet forest. We found that net nitrate (NO3- ) bioavailability is positively related to MAT (r2 =0.79, P=0.0033), and AOA DNA abundance is positively related to both NO3- availability (r2 =0.34, P=0.0071) and MAT (r2 =0.34, P<0.001). In contrast, AOB DNA was only detected in some soils across the gradient. We identified three distinct phylotypes within the AOA which differed from one another in abundance and relative gene expression. In addition, one AOA phylotype increased in abundance with MAT, while others did not. We conclude that MAT is the primary driver of ecosystem N availability across this gradient, and AOA population size and structure appear to mediate the relationship between the nitrification and N bioavailability. These findings hold important implications for nutrient limitation in forests and feedbacks to primary production under changing climate.

Unraveling the active microbial populations involved in nitrogen utilization in a vertical subsurface flow constructed wetland treating urban wastewater.

The dynamics of the active microbial populations involved in nitrogen transformation in a vertical subsurface flow constructed wetland (VF) treating urban wastewater was assessed. The wetland (1.5m2) operated under average loads of 130gCODm-2d-1 and 17gTNm-2d-1 in Period I, and 80gCODm-2d-1 and 19gTNm-2d-1 in Period II. The hydraulic loading rate (HLR) was 375mmd-1 and C/N ratio was 2 in both periods. Samples for microbial characterization were collected from the filter medium (top and bottom layers) of the wetland, water influent and effluent at the end of Periods I (Jun-Oct) and II (Nov-Jan). The combination of qPCR and high-throughput sequencing (NGS, MiSeq) assessment at DNA and RNA level of 16S rRNA genes and nitrogen-based functional genes (amoA and nosZ-clade I) revealed that nitrification was associated both with ammonia-oxidizing bacteria (AOB) (Nitrosospira) and ammonia-oxidizing archaea (AOA) (Nitrososphaeraceae), and nitrite-oxidizing bacteria (NOB) such as Nitrobacter. Considering the active abundance (based in amoA transcripts), the AOA population revealed to be more stable than AOB in both periods and depths of the wetland, being less affected by the organic loading rate (OLR). Although denitrifying bacteria (nosZ copies and transcripts) were actively detected in all depths, the denitrification process was low (removal of 2gTNm-2d-1 for both periods) concomitant with NOx-N accumulation in the effluent. Overall, AOA, AOB and denitrifying bacteria (nosZ) were observed to be more active in bottom than in top layer at lower OLR (Period II). A proper design of OLR and HLR seems to be crucial to control the activity of microbial biofilms in VF wetlands on the basis of oxygen, organic-carbon and NOx-N forms, to improve their capacity for total nitrogen removal.

Variable responses of ammonia oxidizers across soil particle-size fractions affect nitrification in a long-term fertilizer experiment

We used soils from a 33-year fertilizer experiment and performed high-throughput pyrosequencing analyses targeting the ammonia monooxygenase alpha subunit (amoA) gene (also qPCR and terminal restricted fragment length polymorphism (T-RFLP)) to determine whether and how the composition and structure of ammonia-oxidizing microbes in different particle-size fractions (>2000, 2000–200, 200–63, 63–2 and 2–0.1 μm) have changed in response to chronic inorganic (NPK) and organic (NPKM) fertilizer additions under a rice-wheat rotation. We found fertilization and particle-size fractions had strong effects on abundance and structure of ammonia-oxidizers and potential nitrification activity (PNA) (P < 0.05) which was significantly enhanced under NPKM across all soil fractions. PNA was higher in clay fraction and lowest in silt fraction which was largely correlated to total N and C:N ratio. Aggregated boosted tree emphasized the role that total N and C:N ratio play on ammonia oxidizer abundance. Pyrosequencing data revealed that the Nitrososphaera and Nitrosospira were the dominant clusters of ammonia-oxidizing archaea (AOA) and bacteria (AOB), respectively. Even though AOA population was hundreds of times greater than AOB, it seemed AOB responded more sensitively to ambient shifts which was convinced by NMDS analyses based on Bray-Curtis distance of TRFs and also of OTUs from 454 pyrosequencing.

Effects of bioaugmentation in para-nitrophenol-contaminated soil on the abundance and community structure of ammonia-oxidizing bacteria and archaea.

Pseudomonas sp. strain WBC-3 mineralizes the priority pollutant para-nitrophenol (PNP) and releases nitrite (NO2 (-)), which is probably involved in the nitrification. In this study, the rate of PNP removal in soil bioaugmented with strain WBC-3 was more accelerated with more NO2 (-) accumulation than in uninoculated soils. Strain WBC-3 survived well and remained stable throughout the entire period. Real-time polymerase chain reaction (real-time PCR) indicated a higher abundance of ammonia-oxidizing bacteria (AOB) than ammonia-oxidizing archaea (AOA), suggesting that AOB played a greater role in nitrification in the original sampled soil. Real-time PCR and multivariate analysis based on the denaturing gradient gel electrophoresis showed that PNP contamination did not significantly alter the abundance and community structure of ammonia oxidizers except for inhibiting the AOB abundance. Bioaugmentation of PNP-contaminated soil showed a significant effect on AOB populations and community structure as well as AOA populations. In addition, ammonium (NH4 (+)) variation was found to be the primary factor affecting the AOB community structure, as determined by the correlation between the community structures of ammonia oxidizers and environmental factors. It is here proposed that the balance between archaeal and bacterial ammonia oxidation could be influenced significantly by the variation in NH4 (+) levels as caused by bioaugmentation of contaminated soil by a pollutant containing nitrogen.

Long-term fertilization effects on active ammonia oxidizers in an acidic upland soil in China

The effects of long-term fertilization of acidic soils on ammonia-oxidizing archaea (AOA) and bacteria (AOB) communities and its ecological implications remain poorly understood. We chose an acidic upland soil site under long-term (27-year) fertilization to investigate ammonia oxidizer communities under four different regimes: mineral N fertilizer (N), mineral NPK fertilizer (NPK), organic manure (OM) and an unfertilized control (CK). Soil net nitrification rates were significantly higher in OM soils than in CK, N or NPK soils. Quantitative analysis of the distribution of amoA genes by DNA-based stable isotope probing revealed that AOA dominate in CK, N and NPK soils, while AOB dominate in OM soils. Denaturing gradient gel electrophoresis and clone library analyses of amoA genes revealed that Group 1.1a-associated AOA (also referred to as Nitrosotalea) were the most dominant active AOA population (>92%), while Nitrosospira Cluster 3 and Cluster 9 were predominant among active AOB communities. The functional diversity of active ammonia oxidizers in acidic soils is affected by long-term fertilization practices, and the responses of active ammonia oxidizers to mineral fertilizer and organic manure are clearly different. Our results provide strong evidence that AOA are more highly adapted to growth at low pH and low substrate availability than AOB, and they suggest that the niche differentiation and metabolic diversity of ammonia oxidizers in acidic soils are more complex than previously thought.

High abundances of potentially active ammonia-oxidizing bacteria and archaea in oligotrophic, high-altitude lakes of the Sierra Nevada, California, USA.

Nitrification plays a central role in the nitrogen cycle by determining the oxidation state of nitrogen and its subsequent bioavailability and cycling. However, relatively little is known about the underlying ecology of the microbial communities that carry out nitrification in freshwater ecosystems—and particularly within high-altitude oligotrophic lakes, where nitrogen is frequently a limiting nutrient. We quantified ammonia-oxidizing archaea (AOA) and bacteria (AOB) in 9 high-altitude lakes (2289–3160 m) in the Sierra Nevada, California, USA, in relation to spatial and biogeochemical data. Based on their ammonia monooxygenase (amoA) genes, AOB and AOA were frequently detected. AOB were present in 88% of samples and were more abundant than AOA in all samples. Both groups showed >100 fold variation in abundance between different lakes, and were also variable through time within individual lakes. Nutrient concentrations (ammonium, nitrite, nitrate, and phosphate) were generally low but also varied across and within lakes, suggestive of active internal nutrient cycling; AOB abundance was significantly correlated with phosphate (r2 = 0.32, p<0.1), whereas AOA abundance was inversely correlated with lake elevation (r2 = 0.43, p<0.05). We also measured low rates of ammonia oxidation—indicating that AOB, AOA, or both, may be biogeochemically active in these oligotrophic ecosystems. Our data indicate that dynamic populations of AOB and AOA are found in oligotrophic, high-altitude, freshwater lakes.