amoA Gene Copy Numbers Research Articles

Abundance and composition dynamics of soil ammonia-oxidizing archaea in an alpine fir forest on the eastern Tibetan Plateau of China

Real-time qPCR and clone library sequencing targeting amoA genes were used to investigate the seasonal dynamics of an ammonia-oxidizing archaea (AOA) community in an alpine fir forest in western China. AOA were detected at all sampling dates, and there were significant variations in archaeal amoA gene copy numbers (7.63×10(5) to 8.35×10(8) per gram of dry soil) throughout the nongrowing season. Compared with ammonia-oxidizing bacteria (AOB), the AOA displayed a higher abundance on the majority of sampling dates during the freeze-thaw period. All of the AOA sequences fell within soil and sediment lineages and were affiliated with 7 clusters. Compared with the other clusters, cluster 1 was more sensitive to low temperature and was the dominant group in August. In contrast, cluster 3 dominated the AOA community in winter and probably represents a group of cold-adapted archaea. Redundancy analysis (RDA) revealed that the seasonality of the AOA community was mainly attributed to changes in soil temperature and nutrient availability (e.g., dissolved organic nitrogen and carbon). Our results indicate that AOA exist in frozen soils in the alpine coniferous forest ecosystem of the eastern Tibetan Plateau. Moreover, soil temperature may directly and (or) indirectly affect AOA abundance and composition and may further influence the soil N cycle during the winter.

Production Processes Affected Prokaryotic amoA Gene Abundance and Distribution in High-Temperature Petroleum Reservoirs

Although the presence and activity of ammonia-oxidation archaea (AOA) and ammonia-oxidation bacteria (AOB) were observed in thermophilic habitats recently, their existence in the geothermal subterranean oil reservoirs is still not available. This study investigated the abundance and distribution of AOA and AOB in the production waters of high-temperature oil reservoirs by using real-time PCR and phylogenetic analysis based on amoA genes. The results indicated the occurrence of both AOA and AOB in 9 out of totally 17 wells. The AOA-like phylotypes are mainly clustered within two major clades of archaeal amoA sequences known from water columns, sediments and soils: clusters A and B, and a few clones are related to the new genera: Candidatus ‘Nitrosocaldus yellowstonii’. The AOB-like phylotypes mainly belong to Nitrosospira and Nitrosomonas clusters, while two of them are deep-branched in Nitrosospira cluster and showed no substantial alignment to the known cultured AOB, indicating the possibility of new AOB phylotypes. The abundance of AOA and AOB-like amoA genes ranged from 2.92 × 103 to 9.21 × 104 and from 2.91 × 102 to 8.12 × 103 amoA gene copy numbers per ml production water with the ratios of AOA to AOB ranging from 5.10 to 95.5. Statistical analysis showed that amoA gene fell into five groups and the distribution of amoA gene is significantly correlated with the environmental factors, e.g., temperature and recovery process. Our study showed distribution of prokaryotic amoA gene in various oil reservoirs was affected by production processes.

Soil particle surface electrochemical property effects on abundance of ammonia-oxidizing bacteria and ammonia-oxidizing archaea, NH 4+ activity, and net nitrification in an acid soil

Soil surface electrochemical properties may have a strong influence on nitrifying microorganisms, H + and NH 4 + activities, and therefore on the nitrification process. A gradient of surface electrochemical parameters was obtained by amendment of a subtropical acid pine soil (Oxisol) with 0% (control), 3%, 5%, 8%, 10% and 12% pure Ca-Montmorillonite by weight. The H + and NH 4 + activities, the abundance of the ammonia-oxidizing bacterial (AOB) and archaeal (AOA) amoA gene copies, and time-dependent kinetics of net nitrification were investigated. Soil particle surface specific area ranged from 53 to 103 m 2 g −1 and increased with increasing montmorillonite application rate. Similar to specific area, surface charge quantity, surface charge density, electric field strength and surface potential increased after montmorillonite amendment. The H + and NH 4 + activities decreased linearly after montmorillonite addition. AOB amoA gene copy number was 1.82 × 10 5 copies g −1 for unamended soil, and the highest AOB amoA gene copy numbers were found for the 10% montmorillonite amendment (3.11 × 10 7 g −1 soil), which was more than 150 times higher than unamended soil. AOA amoA gene copy numbers were 9.19 × 10 3 copies g −1 dry unamended soil, and the highest AOA amoA gene copy numbers were found in the 8% montmorillonite amendment (1.22 × 10 5 g −1 soil). Although pH significantly decreased during the first three weeks of incubation, no significant difference was observed between the unamended control and different rates of montmorillonite addition treatments during the whole incubation. The largest net nitrification (103 mg N kg −1) was observed in the 10% montmorillonite amendment and the lowest in unamended soil (62 mg N kg −1). While montmorillonite did not change the kinetic patterns of net nitrification, the highest nitrification potential (275 mg N kg −1) for the 10% montmorillonite treatment was more than 3 times higher than unamended soil from simulation of time-dependent kinetics. Nitrification was significantly stimulated after montmorillonite amendment in acid soil mainly due to an increase in the quantity and activity of AOB and AOA. We concluded that soil particle surface parameters can significantly influence nitrification, especially in acid soils.

Long‐Term Field Fertilization Significantly Alters Community Structure of Ammonia‐Oxidizing Bacteria rather than Archaea in a Paddy Soil

Long‐term field fertilization experiments may provide useful insight into the relative contributions of NH3–oxidizing bacteria (AOB) and NH3–oxidizing archaea (AOA) to soil nitrification. In this study, the abundance and composition of AOB and AOA were investigated in bulk soil from paddy field plots (4 by 5 m) that received no fertilization (CK), 180 kg urea N ha−1 yr−1 (NPK), or 180 kg urea N plus 4500 kg rice (Oryza sativa L.) straw ha−1 yr−1 (NPK/OM) since 1988 (each with three replicated plots). Our results clearly indicate that long‐term fertilization (22 yr) significantly altered the community structure of AOB rather than AOA in the soil within fertilized plots compared with the CK plot. Quantitative polymerase chain reaction analysis of the amoA genes that encode the ammonia monooxygenase subunit A revealed that the AOB in the fertilized plots were 50 times more abundant than in the CK plot. Denaturing gradient gel electrophoresis fingerprinting of the amoA genes demonstrated a drastic shift in the AOB community structure, resulting in highly enriched AOB similar to Nitrosospira cluster 3 in the fertilized plots. In contrast, the population size and composition of the AOA community remained largely unchanged in all plots. Furthermore, a positive correlation was observed between the bacterial rather than archaeal amoA gene copy numbers and potential nitrification activity among the soils. The distinct responses of AOA and AOB to long‐term field fertilization were probably related to the availability of NH3. Our results suggest that AOB might play major roles in the NH3 oxidation that occurs in bulk soil under the in situ fertilized paddy field conditions tested in this study.

Effects of nitrogen addition on the abundance and composition of soil ammonia oxidizers in Inner Mongolia Grassland

Nitrogen accumulation in soil is increasing in Inner Mongolia which is resulted mainly from fertilization accompanied by conversion of large area of grasslands to croplands. Ammonia-oxidation is the key step of nitrification which is driven by ammonia-oxidizing microorganisms, and study on the response of ammonia-oxidizing microorganisms is necessary for understanding the effects of nitrogen fertilization on ecosystem functions. In this study, the abundance and community structure of soil ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) under long-term N addition of different rates (0, 1.75, 5.25, 10.5, 17.5, and 28 g N m −2 yr −1) in a typical steppe of the Inner Mongolia Grassland were investigated using quantitative real-time PCR, cloning and sequencing based on amoA gene. In addition, soil potential ammonia oxidation rate was analyzed. Our results demonstrated that, with the increase of nitrogen addition rate, soil pH declined gradually from 6.6 to 4.9, and potential ammonia oxidation rate also declined which was positively correlated with soil pH ( P < 0.01), while the copy number of bacterial amoA gene increased and positively ( P < 0.01) correlated with ammonia concentration in soil. The archaeal amoA gene copy number did not change a lot with N nitrogen addition rate below 10.5 g N/m 2, but significantly decreased with addition of 17.5 and 28 g N m −2 yr −1. Sequencing of clone libraries of treatments revealed that in the treatment without N addition, AOB was dominated by Cluster 3a1 of Nitrosospira with a proportion of 87%, while in the treatment with N addition of 28 g N m −2 yr −1, proportion of Cluster 2 increased significantly to 41%. All archaeal amoA sequences were affiliated with the soil/sediment clade, and no significant variation of community structure was found between the treatments without N addition and with 28 g N m −2 yr −1 addition rate. In conclusion, this study demonstrated significant effects of nitrogen addition on potential ammonia oxidation rate and compositions of ammonia-oxidation microorganisms, which may have important implications for evaluating the impacts of N accumulation on ecosystem functioning. Further, the effects of pH and ammonia concentration on the ammonia oxidation rate and compositions of ammonia-oxidation microorganisms were discussed.

Diversity and quantity of ammonia-oxidizing Archaea and Bacteria in sediment of the Pearl River Estuary, China

The diversity and abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in the sediment of the Pearl River Estuary were investigated by cloning and quantitative real-time polymerase chain reaction (qPCR). From one sediment sample S16, 36 AOA OTUs (3% cutoff) were obtained from three clone libraries constructed using three primer sets for amoA gene. Among the 36 OTUs, six were shared by all three clone libraries, two appeared in two clone libraries, and the other 28 were only recovered in one of the libraries. For AOB, only seven OTUs (based on 16S rRNA gene) and eight OTUs (based on amoA gene) were obtained, showing lower diversity than AOA. The qPCR results revealed that AOA amoA gene copy numbers ranged from 9.6 × 106 to 5.1 × 107 copies per gram of sediment and AOB amoA gene ranged from 9.5 × 104 to 6.2 × 105 copies per gram of sediment, indicating that the dominant ammonia-oxidizing microorganisms in the sediment of the Pearl River Estuary were AOA. The terminal restriction fragment length polymorphism results showed that the relative abundance of AOB species in the sediment samples of different salinity were significantly different, indicating that salinity might be a key factor shaping the AOB community composition.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-011-3107-8) contains supplementary material, which is available to authorized users.

Soil type determines the abundance and community structure of ammonia-oxidizing bacteria and archaea in flooded paddy soils

Ammonia oxidation plays an important role in global nitrogen cycle. However, little information is available on ammonia oxidizers in paddy soils. This study aimed to understand the controlling factors of ammonia oxidizers in the paddy soils. Three types of paddy soils were collected from southwest [Chongqing City (CQ)], central [Honghu City (HH)], and northeast [Panjin City (PJ)] of China, respectively. The soils were cultured with unplanted and rice-planted pot experiments for 10 weeks. The abundance and composition of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) of the paddy soil samples were investigated by real-time polymerase chain reaction and denaturing gradient gel electrophoresis approaches based on amoA genes. In this study, minor changes in the abundance and community structure of both AOB and AOA were observed among the treatments of unplanted and rhizosphere and bulk of rice-planted soil samples in the same type of paddy soil. The AOB population sizes in CQ were lower than those in HH and PJ, while the AOA in CQ and HH were lower than those in PJ. The amoA gene copy numbers of AOA were more abundant than those of AOB in all treatments of the three paddy soils. The community structure of both AOB and AOA in all paddy soils was different from each other. The population size and community structure of AOB and AOA in the flooded paddy soils were mainly determined by the soil types, irrespective of rice-planted or unplanted.

The Influence of Different Land Uses on the Structure of Archaeal Communities in Amazonian Anthrosols Based on 16S rRNA and amoA Genes

Soil from the Amazonian region is usually regarded as unsuitable for agriculture because of its low organic matter content and low pH; however, this region also contains extremely rich soil, the Terra Preta Anthrosol. A diverse archaeal community usually inhabits acidic soils, such as those found in the Amazon. Therefore, we hypothesized that this community should be sensitive to changes in the environment. Here, the archaeal community composition of Terra Preta and adjacent soil was examined in four different sites in the Brazilian Amazon under different anthropic activities. The canonical correspondence analysis of terminal restriction fragment length polymorphisms has shown that the archaeal community structure was mostly influenced by soil attributes that differentiate the Terra Preta from the adjacent soil (i.e., pH, sulfur, and organic matter). Archaeal 16S rRNA gene clone libraries indicated that the two most abundant genera in both soils were Candidatus nitrosphaera and Canditatus nitrosocaldus. An ammonia monoxygenase gene (amoA) clone library analysis indicated that, within each site, there was no significant difference between the clone libraries of Terra Preta and adjacent soils. However, these clone libraries indicated there were significant differences between sites. Quantitative PCR has shown that Terra Preta soils subjected to agriculture displayed a higher number of amoA gene copy numbers than in adjacent soils. On the other hand, soils that were not subjected to agriculture did not display significant differences on amoA gene copy numbers between Terra Preta and adjacent soils. Taken together, our findings indicate that the overall archaeal community structure in these Amazonian soils is determined by the soil type and the current land use.

A lysimeter study of nitrate leaching from grazed grassland as affected by a nitrification inhibitor, dicyandiamide, and relationships with ammonia oxidizing bacteria and archaea

AbstractNitrate (NO3−) can contribute to surface water eutrophication and is deemed harmful to human health if present at high concentrations in the drinking water. In grazed grassland, most of the NO3−‐N leaching occurs from animal urine‐N returns. The objective of this study was to determine the effectiveness of a nitrification inhibitor, dicyandiamide (DCD), in decreasing NO3− leaching in three different soils from different regions of New Zealand under two different rainfall conditions (1260 mm and 2145 mm p.a.), and explore the relationships between NO3−‐N leaching loss and ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA). The DCD nitrification inhibitor was found to be highly effective in decreasing NO3−‐N leaching losses from all three soils under both rainfall conditions. Total NO3−‐N leaching losses from the urine patch areas were decreased from 67.7–457.0 kg NO3−‐N/ha to 29.7–257.4 kg NO3−‐N/ha by the DCD treatment, giving an average decrease of 59%. The total NO3−‐N leaching losses were not significantly affected by the two different rainfall treatments. The total NO3−‐N leaching loss was significantly related to the amoA gene copy numbers of the AOB DNA and to nitrification rate in the soil but not to that of the AOA. These results suggest that the DCD nitrification inhibitor is highly effective in decreasing NO3− leaching under these different soil and rainfall conditions and that the amount of NO3−‐N leached is mainly related to the growth of the AOB population in the nitrogen rich urine patch soils of grazed grassland.

Resistance and resilience of Cu-polluted soil after Cu perturbation, tested by a wide range of soil microbial parameters

Copper (Cu)-polluted and unpolluted soils were used to study the effect of initial pollution on soil biological resistance and resilience by measuring the responses to perturbation using different parameters. Microbial biomass carbon, substrate-induced respiration and copy numbers of 16S rRNA gene were grouped as general parameters, while potential ammonia oxidation rate and copy numbers of amoA gene were grouped as specific functions. In addition, to illustrate how initial pollution affects soil biological resistance and resilience following secondary perturbation, the microbial community structure, together with free Cu(2+) activities ([Cu(2+)]) in soil pore water and soil pH were also measured after secondary perturbation. Results showed that general parameters were more stable than specific ones. High [Cu(2+)] and low pH in soil pore water induced by Cu addition may lead to apparently low resistance and resilience, whereas the formation of a tolerant community after Cu pollution, secondary perturbation and Cu aging may contribute to resistance and resilience. Analysis of the phospholipid fatty acids profile showed that microbial community structure shifted along with the [Cu(2+)] gradient. The microbial community structure of the control soil was both resistant and resilient to 400 mg kg(-1) Cu perturbation, whereas other treatments were neither resistant nor resilient.

Bacteria rather than Archaea dominate microbial ammonia oxidation in an agricultural soil

Agricultural ecosystems annually receive approximately 25% of the global nitrogen input, much of which is oxidized at least once by ammonia-oxidizing prokaryotes to complete the nitrogen cycle. Recent discoveries have expanded the known ammonia-oxidizing prokaryotes from the domain Bacteria to Archaea. However, in the complex soil environment it remains unclear whether ammonia oxidation is exclusively or predominantly linked to Archaea as implied by their exceptionally high abundance. Here we show that Bacteria rather than Archaea functionally dominate ammonia oxidation in an agricultural soil, despite the fact that archaeal versus bacterial amoA genes are numerically more dominant. In soil microcosms, in which ammonia oxidation was stimulated by ammonium and inhibited by acetylene, activity change was paralleled by abundance change of bacterial but not of archaeal amoA gene copy numbers. Molecular fingerprinting of amoA genes also coupled ammonia oxidation activity with bacterial but not archaeal amoA gene patterns. DNA-stable isotope probing demonstrated CO(2) assimilation by Bacteria rather than Archaea. Our results indicate that Archaea were not important for ammonia oxidation in the agricultural soil tested.

Growth of ammonia-oxidizing archaea in soil microcosms is inhibited by acetylene

Autotrophic ammonia-oxidizing bacteria were considered to be responsible for the majority of ammonia oxidation in soil until the recent discovery of the autotrophic ammonia-oxidizing archaea. To assess the relative contributions of bacterial and archaeal ammonia oxidizers to soil ammonia oxidation, their growth was analysed during active nitrification in soil microcosms incubated for 30 days at 30 degrees C, and the effect of an inhibitor of ammonia oxidation (acetylene) on their growth and soil nitrification kinetics was determined. Denaturing gradient gel electrophoresis (DGGE) analysis of bacterial ammonia oxidizer 16S rRNA genes did not detect any change in their community composition during incubation, and quantitative PCR (qPCR) analysis of bacterial amoA genes indicated a small decrease in abundance in control and acetylene-containing microcosms. DGGE fingerprints of archaeal amoA and 16S rRNA genes demonstrated changes in the relative abundance of specific crenarchaeal phylotypes during active nitrification. Growth was also indicated by increases in crenarchaeal amoA gene copy number, determined by qPCR. In microcosms containing acetylene, nitrification and growth of the crenarchaeal phylotypes were suppressed, suggesting that these crenarchaea are ammonia oxidizers. Growth of only archaeal but not bacterial ammonia oxidizers occurred in microcosms with active nitrification, indicating that ammonia oxidation was mostly due to archaea in the conditions of the present study.

Bacteria, not archaea, restore nitrification in a zinc-contaminated soil

Biological ammonia oxidation had long been thought to be mediated solely by discrete clades of beta- and gamma-proteobacteria (ammonia-oxidizing bacteria; AOB). However, ammonia-oxidizing Crenarchaeota (ammonia-oxidizing archaea; AOA) have recently been identified and proposed to be the dominant agents of ammonia oxidation in soils. Nevertheless, the dynamics of AOB versus AOA, and their relative contribution to soil ammonia oxidation and ecosystem functioning on stress and environmental perturbation, remain unknown. Using a 3-year longitudinal field study and the amoA gene as a molecular marker, we demonstrate that AOB, but not AOA, mediate recovery of nitrification after zinc (Zn) contamination. Pristine soils showed approximately equal amoA gene copy numbers and transcript levels for AOB and AOA. At an intermediate Zn dose (33.7 mmol Zn per kg), ammonia oxidation was completely inhibited, and the numbers of AOB and AOA amoA gene copies and gene transcripts were reduced. After 2 years, ammonia oxidation in the field soils was fully restored to preexposure levels, and this restoration of function was concomitant with an increase of AOB amoA gene copy and gene transcript numbers. Analysis of the restored community revealed domination by a phylogenetically distinct Zn-tolerant Nitrosospira sp. community. In contrast, the numbers of AOA amoA gene copies and gene transcripts remained 3- and 10(4)-fold lower than recovered AOB values, respectively. Thus, although recent findings have emphasized a dominant role of archaea in soil-borne ammonia oxidation, we demonstrate that a phylogenetic shift within the AOB community drives recovery of nitrification from Zn contamination in this soil.

Major gradients in putatively nitrifying and non-nitrifying Archaea in the deep North Atlantic

Aerobic nitrification of ammonia to nitrite and nitrate is a key process in the oceanic nitrogen cycling mediated by prokaryotes. Apart from Bacteria belonging to the beta- and gamma-Proteobacteria involved in the first nitrification step, Crenarchaeota have recently been recognized as main drivers of the oxidation of ammonia to nitrite in soil as well as in the ocean, as indicated by the dominance of archaeal ammonia monooxygenase (amoA) genes over bacterial amoA. Evidence is accumulating that archaeal amoA genes are common in a wide range of marine systems. Essentially, all these reports focused on surface and mesopelagic (200-1,000 m depth) waters, where ammonia concentrations are higher than in waters below 1,000 m depth. However, Crenarchaeota are also abundant in the water column below 1,000 m, where ammonia concentrations are extremely low. Here we show that, throughout the North Atlantic Ocean, the abundance of archaeal amoA genes decreases markedly from subsurface waters to 4,000 m depth, and from subpolar to equatorial deep waters, leading to pronounced vertical and latitudinal gradients in the ratio of archaeal amoA to crenarchaeal 16S ribosomal RNA (rRNA) genes. The lack of significant copy numbers of amoA genes and the very low fixation rates of dark carbon dioxide in the bathypelagic North Atlantic suggest that most bathypelagic Crenarchaeota are not autotrophic ammonia oxidizers: most likely, they utilize organic matter and hence live heterotrophically.

The effect of hydroxylamine on the activity and aggregate structure of autotrophic nitrifying bioreactor cultures

Addition of hydroxylamine (NH2OH) to autotrophic biomass in nitrifying bioreactors affected the activity, physical structure, and microbial ecology of nitrifying aggregates. When NH2OH is added to nitrifying cultures in 6-h batch experiments, the initial NH3-N uptake rates were physiologically accelerated by a factor of 1.4-13. NH2OH addition caused a 20-40% decrease in the median aggregate size, broadened the shape of the aggregate size distribution by up to 230%, and caused some of the microcolonies to appear slightly more dispersed. Longer term NH2OH addition in fed batch bioreactors decreased the median aggregate size, broadened the aggregate size distribution, and decreased NH3-N removal from >90% to values ranging between 75% and 17%. This altered performance is explained by quantitative fluorescence in situ hybridization (FISH) results that show inhibition of nitrifying populations, and by qPCR results showing that the copy numbers of amoA and nxrA genes gradually decreased by up to an order-of-magnitude. Longer term NH2OH addition damaged the active biomass. This research clarifies the effect of NH2OH on nitrification and demonstrates the need to incorporate NH2OH-related dynamics of the nitrifying biomass into mathematical models, accounting for both ecophysiological and structural responses.

Spatial distribution of Bacteria and Archaea and amoA gene copy numbers throughout the water column of the Eastern Mediterranean Sea

Until recently, ammonia oxidation, a key process in the global nitrogen cycle, was thought to be mediated exclusively by a few bacterial groups. It has been shown now, that also Crenarchaeota are capable to perform this initial nitrification step. The abundance of ammonia oxidizing Bacteria and Archaea was determined using the bacterial and archaeal ammonia monooxygenase-alpha subunit (amoA) gene as functional markers in a quantitative PCR approach and related to the abundance of Bacteria and Archaea in the Eastern Mediterranean Sea. Archaeal amoA copy numbers decreased from 4000-5000 copies ml(-1) seawater from the 200-500 m depth layer to 20 copies ml(-1) at 1000 m depth. Beta-proteobacterial amoA genes were below the detection limit in all the samples. The archaeal amoA copy numbers were correlated with NO(2)(-) concentrations, suggesting that ammonia-oxidizing Archaea may play a significant role in the nitrification in the mesopelagic waters of the Eastern Mediterranean Sea. In the bathypelagic waters, however, archaeal amoA gene abundance was rather low although Crenarchaeota were abundant, indicating that Crenarchaeota might largely lack the amoA gene in these deep waters. Terminal restriction fragment length polymorphism analysis of the archaeal community revealed a distinct clustering with the mesopelagic community distinctly different from the archaeal communities of both, the surface waters and the 3000-4000 m layers. Hence, the archaeal community in the Eastern Mediterranean Sea appears to be highly stratified despite the absence of major temperature and density gradients between the meso- and bathypelagic waters of the Mediterranean Sea.

Abundance and composition of ammonia‐oxidizing bacteria and ammonia‐oxidizing archaea communities of an alkaline sandy loam

The abundance and composition of soil ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) communities under different long-term (17 years) fertilization practices were investigated using real-time polymerase chain reaction and denaturing gradient gel electrophoresis (DGGE). A sandy loam with pH (H(2)O) ranging from 8.3 to 8.7 was sampled in years 2006 and 2007, including seven fertilization treatments of control without fertilizers (CK), those with combinations of fertilizer nitrogen (N), phosphorus (P) and potassium (K): NP, NK, PK and NPK, half chemical fertilizers NPK plus half organic manure (1/2OMN) and organic manure (OM). The highest bacterial amoA gene copy numbers were found in those treatments receiving N fertilizer. The archaeal amoA gene copy numbers ranging from 1.54 x 10(7) to 4.25 x 10(7) per gram of dry soil were significantly higher than those of bacterial amoA genes, ranging from 1.24 x 10(5) to 2.79 x 10(6) per gram of dry soil, which indicated a potential role of AOA in nitrification. Ammonia-oxidizing bacteria abundance had significant correlations with soil pH and potential nitrification rates. Denaturing gradient gel electrophoresis patterns revealed that the fertilization resulted in an obvious change of the AOB community, while no significant change of the AOA community was observed among different treatments. Phylogenetic analysis showed a dominance of Nitrosospira-like sequences, while three bands were affiliated with the Nitrosomonas genus. All AOA sequences fell within cluster S (soil origin) and cluster M (marine and sediment origin). These results suggest that long-term fertilization had a significant impact on AOB abundance and composition, while minimal on AOA in the alkaline soil.

Quantitative analyses of the abundance and composition of ammonia‐oxidizing bacteria and ammonia‐oxidizing archaea of a Chinese upland red soil under long‐term fertilization practices

The abundance and composition of soil ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) were investigated by using quantitative real-time polymerase chain reaction, cloning and sequencing approaches based on amoA genes. The soil, classified as agri-udic ferrosols with pH (H(2)O) ranging from 3.7 to 6.0, was sampled in summer and winter from long-term field experimental plots which had received 16 years continuous fertilization treatments, including fallow (CK0), control without fertilizers (CK) and those with combinations of fertilizer nitrogen (N), phosphorus (P) and potassium (K): N, NP, NK, PK, NPK and NPK plus organic manure (OM). Population sizes of AOB and AOA changed greatly in response to the different fertilization treatments. The NPK + OM treatment had the highest copy numbers of AOB and AOA amoA genes among the treatments that received mineral fertilizers, whereas the lowest copy numbers were recorded in the N treatment. Ammonia-oxidizing archaea were more abundant than AOB in all the corresponding treatments, with AOA to AOB ratios ranging from 1.02 to 12.36. Significant positive correlations were observed among the population sizes of AOB and AOA, soil pH and potential nitrification rates, indicating that both AOB and AOA played an important role in ammonia oxidation in the soil. Phylogenetic analyses of the amoA gene fragments showed that all AOB sequences from different treatments were affiliated with Nitrosospira or Nitrosospira-like species and grouped into cluster 3, and little difference in AOB community composition was recorded among different treatments. All AOA sequences fell within cluster S (soil origin) and cluster M (marine and sediment origin). Cluster M dominated exclusively in the N, NP, NK and PK treatments, indicating a pronounced difference in the community composition of AOA in response to the long-term fertilization treatments. These findings could be fundamental to improve our understanding of the importance of both AOB and AOA in the cycling of nitrogen and other nutrients in terrestrial ecosystems.

Quantitative distribution of presumptive archaeal and bacterial nitrifiers in Monterey Bay and the North Pacific Subtropical Gyre

The recent isolation of the ammonia-oxidizing crenarchaeon Nitrosopumilus maritimus has expanded the known phylogenetic distribution of nitrifying phenotypes beyond the domain Bacteria. To further characterize nitrification in the marine environment and explore the potential crenarchaeal contribution to this process, we quantified putative nitrifying genes and phylotypes in picoplankton genomic libraries and environmental DNA samples from coastal and open ocean habitats. Betaproteobacteria ammonia monooxygenase subunit A (amoA) gene copy numbers were low or undetectable, in stark contrast to crenarchaeal amoA-like genes that were broadly distributed and reached up to 6 x 10(4) copies ml(-1). Unexpectedly, in the North Pacific Subtropical Gyre, a deeply branching crenarchaeal group related to a hot spring clade (pSL12) was at times abundant below the euphotic zone. Quantitative data suggested that the pSL12 relatives also contain archaeal amoA-like genes. In both coastal and open ocean habitats, close relatives of known nitrite-oxidizing Nitrospina species were well represented in genomic DNA libraries and quantitative PCR profiles. Planktonic Nitrospina depth distributions correlated with those of Crenarchaea. Overall, the data suggest that amoA-containing Crenarchaea are more phylogenetically diverse than previously reported. Additionally, distributional patterns of planktonic Crenarchaea and Nitrospina species suggest potential metabolic interactions between these groups in the ocean's water column.

Quantification of Ammonia-Oxidizing Bacteria and Factors Controlling Nitrification in Salt Marsh Sediments

To elucidate the geomicrobiological factors controlling nitrification in salt marsh sediments, a comprehensive approach involving sediment geochemistry, process rate measurements, and quantification of the genetic potential for nitrification was applied to three contrasting salt marsh habitats: areas colonized by the tall (TS) or short (SS) form of Spartina alterniflora and unvegetated creek banks (CBs). Nitrification and denitrification potential rates were strongly correlated with one another and with macrofaunal burrow abundance, indicating that coupled nitrification-denitrification was enhanced by macrofaunal burrowing activity. Ammonia monooxygenase (amoA) gene copy numbers were used to estimate the ammonia-oxidizing bacterial population size (5.6 x 10(4) to 1.3 x 10(6) g of wet sediment(-1)), which correlated with nitrification potentials and was 1 order of magnitude higher for TS and CB than for SS. TS and CB sediments also had higher Fe(III) content, higher Fe(III)-to-total reduced sulfur ratios, higher Fe(III) reduction rates, and lower dissolved sulfides than SS sediments. Iron(III) content and reduction rates were positively correlated with nitrification and denitrification potential and amoA gene copy number. Laboratory slurry incubations supported field data, confirming that increased amounts of Fe(III) relieved sulfide inhibition of nitrification. We propose that macrofaunal burrowing and high concentrations of Fe(III) stimulate nitrifying bacterial populations, and thus may increase nitrogen removal through coupled nitrification-denitrification in salt marsh sediments.