Ammonia-oxidizing Bacteria Community Composition Research Articles

Influence of the nitrification inhibitor DMPP on the community composition of ammonia-oxidizing bacteria at microsites with increasing distance from the fertilizer zone

The effect of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) on N transformations and composition of ammonia-oxidizing bacteria (AOB) communities was investigated at the centimeter scale in a microcosm experiment under laboratory conditions. After 28 days, samples were collected from soil treated with urea or urea and DMPP at increasing distance from the fertilizer zone; this distance ranged from 0 to 5 cm in both horizontal and vertical directions. The results showed that DMPP application significantly increased soil pH and NH4+-N and mineral N (NH4+-N, NO3−-N, and NO2−-N) concentrations but decreased (NO3− + NO2−)-N concentration, and such effect was decreased by increasing the distance from the fertilizer zone. Fingerprint profiles of denaturing gradient gel electrophoresis showed that the number of bands decreased by increasing the distance from the fertilizer zone due to decreasing NH4+-N concentrations in the urea treatment. Compared to urea applied alone, DMPP application increased NH4+-N concentrations and decreased AOB diversity from 0 to 3 cm but promoted diversity from 3 to 5 cm distance from the fertilizer zone. A phylogenetic analysis showed that AOB communities were dominated by Nitrosospira cluster 3. Therefore, the nitrification inhibitor DMPP modified the composition of AOB communities by increasing the distance from the fertilizer zone and this probably was related to the changes in soil pH and inorganic N concentration.

Responses of bacterial and archaeal ammonia oxidisers of an acidic luvisols soil to different nitrogen fertilization rates after 9 years

It is still not clear which group of ammonia-oxidizing microorganisms plays the most important roles in nitrification in soils. Change in abundances and community compositions of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) under long-term different nitrogen (N) fertilization rates were investigated in an acidic luvisols soil using real-time polymerase chain reaction and denaturing gradient gel electrophoresis, respectively, based on the ammonia monooxygenase a-subunit gene. The experimental plan included the following treatments: control without N fertilization (NCK), low N fertilization rate, middle N fertilization rate, and high N fertilization rate as 0, 100, 150, and 250 kg urea-N ha−1, respectively. Long-term different N fertilization rates did not significantly alter the total C and N contents of soil while it significantly decreased soil pH, which ranged from 5.60 to 5.20. The AOB abundance was more abundant in the N fertilization treatments than the NCK treatment; the AOA abundance decreased by the increasing N fertilization rates, as did the ratios of AOA/AOB. The large differences in the potential nitrification rates among four treatments depended on the changes in AOA abundance but not to changes in AOB abundance. Phylogenetic analysis showed that the AOB communities were dominated by Nitrosospira clusters 1, 3, and 9 while all AOA sequences were grouped into soil/sediment cluster except for one sequence. Taken together, these results indicated that AOB and AOA preferred different soil N conditions and AOA were functionally more important in the nitrification than AOB in the acidic luvisols soil.

Relationship between ammonia oxidizing bacteria and bioavailable nitrogen in harvested forest soils of central Alberta

Forest soils are commonly limited in nitrogen (N), and the removal of aboveground biomass in harvesting operations can exacerbate the problem. Thus, the soil organisms that facilitate the rate-limiting step in the N cycle, the oxidation of ammonium (NH 4 +), are of special interest in harvested environments. The objective of this study was to investigate the changes in ammonia oxidizing bacteria (AOB) communities that occurred in the years following clear cutting, and link those community shifts to availability of inorganic N forms NH 4 + and nitrate (NO 3 −). Genetic fingerprinting targeting the amoA gene coupled with denaturing gel gradient electrophoresis was carried out over two summers on forest floor (LFH) and mineral (Ae) soils of three similar cutblocks harvested during different years. In- situ NH 4 + and NO 3 − availability was measured over the growing seasons of 2009 and 2010, as well as a suite of physical soil characteristics. Results indicated that the AOB community composition differed in younger vs. older cutblocks, but not by soil horizon. The changes seen in the AOB paralleled the change in N bioavailability across sites, soil horizons, and sampling years, thus indicating that N bioavailability may be directly linked to AOB community composition. This link may provide the basis for the use of AOB as indicators of nutrient availability in the future.

Nitrogen Fertilization Changes Abundance and Community Composition of Ammonia-Oxidizing Bacteria

The effects of N fertilization rates on the abundance and community composition of NH3–oxidizing bacteria (AOB) are poorly understood, particularly in greenhouse-based high-N-input vegetable soils. The abundance and community composition of AOB from these vegetable soils under five N (urea) rates in eastern China were evaluated by polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE). These rates were: the conventional high N rate (N4) as 270 or 300 kg N ha−1 season−1 for cucumber (Cucumis sativus L.) or tomato (Solanum lycopersicum L.), respectively, and 80% (N3), 60% (N2), 40% (N1), and 0% (N0) of the N4 rate. After 2 yr of a cucumber–tomato rotation, N fertilization had significant effects on the abundance and community composition of AOB. The abundance of AOB amoA gene was significantly higher under the N2 (2.08 × 1010 kg−1 dry weight soil) than under the N0 or N4 (8.74 or 8.75 × 109 kg−1 soil). The DGGE patterns were more diverse under N1 and N2 than under N3 and N4, with the greatest DGGE band numbers (15) under N2. Phylogenetic analyses of amoA gene fragments showed that AOB sequences from different N fertilization rates were affiliated with Nitrosospira species and grouped into Clusters 2, 3, 8, and 12. Our results indicate that a reduction to 40% of the conventional high N rate (N4) is practical to reduce N inputs while maintaining the abundance and community composition of AOB in such greenhouse-based vegetable soils.

Bacterial and archaeal amoA gene distribution covaries with soil nitrification properties across a range of land uses

Ammonia-oxidizing bacteria and ammonia-oxidizing archaea are commonly found together in soils, yet the factors influencing their relative distribution and activity remain unclear. We examined archaeal and bacterial amoA gene distribution, and used a novel bioassay to assess archaeal and bacterial contributions to nitrification potentials in soils spanning a range of land uses (forest, pasture, cultivated and long-term fallowed cropland) along a 10 km transect. The assay, which quantifies the extent to which acetylene-inactivated soil nitrification potential recovers (RNP) in the presence of bacterial protein synthesis inhibitors, indicated a significant archaeal contribution to the nitrification potentials of the pasture and long-term fallowed soils. Archaeal amoA gene abundance did not vary significantly among the soils, but bacterial amoA gene abundance did, resulting in archaeal : bacterial amoA abundance ratios ranging from 1.1 ± 0.8 in cultivated soils to 396 ± 176 in pasture soils. Both archaeal and bacterial amoA gene compositions were heterogeneous across the landscape, but differed in their patterns of variability. Archaeal amoA gene distributions were distinct among each of the three main land-use types: forest, pasture and cropland soils. In contrast, bacterial amoA gene composition was distinct in forest and in cultivated cropland, while pasture and long-term fallowed cropland soils were similar. In both pasture and long-term fallowed cropland soils, one phylotype of Nitrosospira cluster 3a was highly abundant. This distinct bacterial amoA gene fingerprint correlated with significant contributions of archaea to RNP of both soils, despite differences in archaeal amoA gene composition between the pasture and fallowed soils. This observation suggests that the factors driving the development of ammonia-oxidizing bacteria community composition might influence the extent of archaeal contribution to soil nitrification.

Influence of operational parameters on nitrogen removal efficiency and microbial communities in a full-scale activated sludge process

To improve the efficiency of total nitrogen (TN) removal, solid retention time (SRT) and internal recycling ratio controls were selected as operating parameters in a full-scale activated sludge process treating high strength industrial wastewater. Increased biomass concentration via SRT control enhanced TN removal. Also, decreasing the internal recycling ratio restored the nitrification process, which had been inhibited by phenol shock loading. Therefore, physiological alteration of the bacterial populations by application of specific operational strategies may stabilize the activated sludge process. Additionally, two dominant ammonia oxidizing bacteria (AOB) populations, Nitrosomonas europaea and Nitrosomonas nitrosa, were observed in all samples with no change in the community composition of AOB. In a nitrification tank, it was observed that the Nitrobacter populations consistently exceeded those of the Nitrospira within the nitrite oxidizing bacteria (NOB) community. Through using quantitative real-time PCR (qPCR), nirS, the nitrite reducing functional gene, was observed to predominate in the activated sludge of an anoxic tank, whereas there was the least amount of the narG gene, the nitrate reducing functional gene.

Nitrification and nitrifying bacteria in the Chengdu section of middle Min River (China)

The Min River is one of the most important tributaries of the upper Yangtze River. Its middle reach, below Chengdu city, receives large numbers of effluents from Sanwayao wastewater treatment plant (WWTP), where ammonia concentration raises significantly. In this study, the situation of ammonia pollution, nitrification and nitrifying bacteria in the Min River was studied during the year 2007 to 2008. Nitrifying activity was assessed under optimum conditions based on the background ammonia concentration and abundance of nitrifying bacteria was measured by the real time quantitative-polymerase chain reaction (RTQ-PCR). The community composition of ammonia-oxidizing bacteria (AOB) was investigated with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S ribosomal DNA fragments. The results showed that the ammonia contamination of the Chengdu section of the middle Min River was severe. Nitrification mainly takes place in the 30 km reach of the lower river, where the numbers of nitrifying bacteria are relatively more than those in the upper river do. The sequence analysis of the DGGE bands reveals that the majority of AOB presenting in the river belong to lineage 2, represented by Nitrosomonas oligotropha-and Nitrosomonas ureae-like bacteria. The rest of the AOB population is represented in minor proportions by Nitrosospira-like species.

Rapid and dissimilar response of ammonia oxidizing archaea and bacteria to nitrogen and water amendment in two temperate forest soils

Biochemical processes relevant to soil nitrogen (N) cycling are performed by soil microorganisms affiliated with diverse phylogenetic groups. For example, the oxidation of ammonia, representing the first step of nitrification, can be performed by ammonia oxidizing bacteria (AOB) and, as recently reported, also by ammonia oxidizing archaea (AOA). However, the contribution to ammonia oxidation of the phylogenetically separated AOA versus AOB and their respective responsiveness to environmental factors are still poorly understood. The present study aims at comparing the capacity of AOA and AOB to momentarily respond to N input and increased soil moisture in two contrasting forest soils. Soils from the pristine Rothwald forest and the managed Schottenwald forest were amended with either NH 4 +–N or NO 3 −–N and were incubated at 40% and 70% water-filled pore space (WFPS) for four days. Nitrification rates were measured and AOA and AOB abundance and community composition were determined via quantitative PCR (qPCR) and terminal restriction length fragment polymorphism (T-RFLP) analysis of bacterial and archaeal amoA genes. Our study reports rapid and distinct changes in AOA and AOB abundances in the two forest soils in response to N input and increased soil moisture but no significant effects on net nitrification rates. Functional microbial communities differed significantly in the two soils and responded specifically to the treatments during the short-term incubation. In the Rothwald soil the abundance and community composition of AOA were affected by the water content, whereas AOB communities responded to N amendment. In the Schottenwald soil, by contrast, AOA responded to N addition. These results suggest that AOA and AOB may be selectively influenced by soil and management factors.

Impacts of Spartina alterniflora invasion on abundance and composition of ammonia oxidizers in estuarine sediment

Spartina alterniflora widely invades coastal wetland in China and might change nitrification in sediment. Both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) are involved in nitrification in this environment. The objective of this study was to examine the effect of S. alterniflora invasion on abundance and composition of AOA and AOB. The abundance and composition of AOA, AOB, and total bacteria in the sediments from S. alterniflora-invaded native mangrove vegetated and unvegetated zones at two depths of 0–5 cm (O) and 5–20 cm (R) were investigated using quantitative real-time polymerase chain reaction and denaturing gradient gel electrophoresis. Relationships were also determined between sediment properties and the AOB and AOA population sizes. Compared with native mangrove vegetated zone, the archaeal amoA gene abundance was reduced by 11.3-fold (O) and 46.1-fold (R), but the bacterial amoA gene abundance was increased by 9.8-fold (O) and 1.8-fold (R), respectively, in the S. alterniflora-invaded zone. The AOA abundance was always higher than AOB, especially in the native mangrove zone. Both AOA and AOB population sizes in the upper layer (O) were bigger than those in the deeper layer (R). Little difference was found in the AOB community composition among different zones, while diversity of AOA community was increased by the presence of S. alterniflora. This study demonstrated that the S. alterniflora invasion affects the abundance of both AOA and AOB, but only affects the community composition of AOA in the tidal sediments.

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.

Abundance and community structure of ammonia-oxidizing archaea and bacteria in an acid paddy soil

Nitrification is essential to the nitrogen cycle in paddy soils. However, it is still not clear which group of ammonia-oxidizing microorganisms plays more important roles in nitrification in the paddy soils. The changes in the abundance and composition of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) were investigated by real-time PCR, terminal restriction fragment length polymorphism, and clone library approaches in an acid red paddy soil subjected to long-term fertilization treatments, including treatment without fertilizers (CT); chemical fertilizer nitrogen (N); N and potassium (NK); N and phosphorus (NP); N, P, and K (NPK); and NPK plus recycled crop residues (NPK+C). The AOA population size in NPK+C was higher than those in CT, while minor changes in AOB population sizes were detected among the treatments. There were also some changes in AOA community composition responding to different fertilization treatments. Still few differences were detected in AOB community composition among the treatments. Phylogenetic analysis showed that the AOA sequences fell into two main clusters: cluster A and cluster soil/sediment. The AOB composition in this paddy soil was dominated by Nitrosospira cluster 12. These results suggested that the AOA were more sensitive than AOB to different fertilization treatments in the acid red paddy soil.

Influence of bacterial-feeding nematodes on nitrification and the ammonia-oxidizing bacteria (AOB) community composition

The effects of bacterial-feeding nematodes on nitrification and the ammonia-oxidizing bacteria (AOB) community composition were studied in soil microcosms. Sterilized soils were inoculated with mixed soil bacteria (obtained by filtering) or with bacteria and bacterial-feeding nematodes, after which the dynamic inorganic nitrogen concentration was measured weekly. After 28 days of incubation, denaturing gradient gel electrophoresis (DGGE) based on PCR amplification of the amoA gene was used to analyze the AOB community composition. In addition, a clone library from the amoA gene fragments was established using clones randomly selected and sequenced from the two treatments. The results showed that the presence of bacterial-feeding nematodes led to significantly greater NH 4 + and NO 3 − contents over the entire incubation period, indicating that bacterial-feeding nematodes promoted both N mineralization and nitrification. The results of DGGE showed that the AOB community composition was significantly changed in the presence of bacterial-feeding nematodes. Furthermore, the sequencing results suggested that Nitrosospira sp. was the dominant species in the treatment without nematodes, while Nitrosomonas sp. and Nitrosospira sp. were the dominant species in the treatment with nematodes. Such changes in the AOB community may be one of explanation of the important role that nematodes play in promoting nitrification.

Effect of common bean ( Phaseolus vulgaris L.) on the community composition of ammonia-oxidizing bacteria in soil previously cultivated with Medicago sativa

The community composition of ammonia-oxidizing bacteria (AOB) was studied during four stages of plant development in soil cultivated with Phaseolus vulgaris in comparison to unplanted soil, using an alfisol previously harboring the legume Medicago sativa. Denaturing gradient gel electrophoresis (DGGE) patterns of 16S rRNA gene and clone libraries of the same gene suggested that bacteria related to Nitrosospira cluster 3 were dominant in both planted and unplanted soil. Bacteria related to Nitrosomonas cluster 8 (Nitrosomonas communis cluster) were found at all times in planted soil, but appeared only randomly in unplanted soil. Analysis of PCR products of the gene encoding the alpha-subunit of ammonia monooxygenase (amoA) by DGGE and clone libraries only detected Nitrosospira cluster 3-like organisms, but failed to detect sequences related to Nitrosomonas. The results suggest that P. vulgaris does not affect the dominant members of AOB communities (Nitrosospira cluster 3), but could have an effect on the prevalence of Nitrosomonas cluster 8 in this type of legume-planted alfisol.

Community composition of ammonia-oxidizing bacteria and archaea in rice field soil as affected by nitrogen fertilization

Little information is available on the ecology of ammonia-oxidizing bacteria (AOB) and archaea (AOA) in flooded rice soils. Consequently, a microcosm experiment was conducted to determine the effect of nitrogen fertilizer on the composition of AOB and AOA communities in rice soil by using molecular analyses of ammonia monooxygenase gene ( amoA) fragments. Experimental treatments included three levels of N (urea) fertilizer, i.e. 50, 100 and 150 mg N kg −1 soil. Soil samples were operationally divided into four fractions: surface soil, bulk soil deep layer, rhizosphere and washed root material. NH 4 +-N was the dominant form of N in soil porewater and increased with N fertilization. Cloning and sequencing of amoA gene fragments showed that the AOB community in the rice soil consisted of three major groups, i.e. Nitrosomonas communis cluster, Nitrosospira cluster 3a and cluster 3b. The sequences related to Nitrosomonas were predominant. There was a clear effect of N fertilizer and soil depth on AOB community composition based on terminal restriction fragment length polymorphism fingerprinting. Nitrosomonas appeared to be more abundant in the potentially oxic or micro-oxic fractions, including surface soil, rhizosphere and washed root material, than the deep layer of anoxic bulk soil. Furthermore, Nitrosomonas increased relatively in the partially oxic fractions and that of Nitrosospira decreased with the increasing application of N fertilizer. However, AOA community composition remained unchanged according to the denaturing gradient gel electrophoresis analyses.

Niche separation of ammonia‐oxidizing bacteria across a tidal freshwater marsh

Like many functional groups or guilds of microorganisms, the group of ammonia-oxidizing bacteria (AOB) consists of a number of physiologically different species or lineages. These physiological differences suggest niche differentiation among these bacteria depending on the environmental conditions. Species of AOB might be adapted to different zones in the flooding gradient of a tidal marsh. This issue has been studied by sampling sediments from different sites and depths within a tidal freshwater marsh along the river Scheldt near the village of Appels in Belgium. Samples were taken in February, April, July and October 1998. Communities of AOB in the sediment were analysed on the basis of the 16S rRNA gene by application of polymerase chain reaction in combination with denaturing gradient gel electrophoresis (DGGE). In addition, moisture content and concentrations of ammonium and nitrate were determined as well as the potential ammonia-oxidizing activities. Six different DGGE bands belonging to the beta-subclass of the Proteobacteria were observed across the marsh. The community composition of AOB was determined by the elevation in the flooding gradient as well as by the sampling depth. The presence of plants was less important for the community composition of AOB. DGGE bands affiliated with the Nitrosospira lineage were mostly found in the upper part of the marsh and in the deeper layers of the sediment. Two of the three DGGE bands related to the Nitrosomonas oligotropha lineage were more broadly distributed over the marsh, but were predominantly found in the upper layers of the sediment. Members of the environmental Nitrosomonas lineage 5 were predominantly detected in the deeper layers in the lower parts of the marsh. Potential driving factors for niche differentiation are discussed.

Community composition of ammonia‐oxidizing bacteria and archaea in soils under stands of red alder and Douglas fir in Oregon

This study determined nitrification activity and nitrifier community composition in soils under stands of red alder (Alnus rubra) and Douglas fir (Pseudotsuga menziesii) at two sites in Oregon. The H.J. Andrews Experimental Forest, located in the Cascade Mountains of Oregon, has low net N mineralization and gross nitrification rates. Cascade Head Experimental Forest, in the Coast Range, has higher net N mineralization and nitrification rates and soil pH is lower. Communities of putative bacterial [ammonia-oxidizing bacteria (AOB)] and archaeal [ammonia-oxidizing archaea (AOA)] ammonia oxidizers were examined by targeting the gene amoA, which codes for subunit A of ammonia monooxygenase. Nitrification potential was significantly higher in red alder compared with Douglas-fir soil and greater at Cascade Head than H.J. Andrews. Ammonia-oxidizing bacteria amoA genes were amplified from all soils, but AOA amoA genes could only be amplified at Cascade Head. Gene copy numbers of AOB and AOA amoA were similar at Cascade Head regardless of tree type (2.3-6.0 x 10(6)amoA gene copies g(-1) of soil). DNA sequences of amoA revealed that AOB were members of Nitrosospira clusters 1, 2 and 4. Ammonia-oxidizing bacteria community composition, determined by terminal restriction fragment length polymorphism (T-RFLP) profiles, varied among sites and between tree types. Many of the AOA amoA sequences clustered with environmental clones previously obtained from soil; however, several sequences were more similar to clones previously recovered from marine and estuarine sediments. As with AOB, the AOA community composition differed between red alder and Douglas-fir soils.

Epiphyton as a Niche for Ammonia-Oxidizing Bacteria: Detailed Comparison with Benthic and Pelagic Compartments in Shallow Freshwater Lakes

Next to the benthic and pelagic compartments, the epiphyton of submerged macrophytes may offer an additional niche for ammonia-oxidizing bacteria in shallow freshwater lakes. In this study, we explored the potential activities and community compositions of ammonia-oxidizing bacteria of the epiphytic, benthic, and pelagic compartments of seven shallow freshwater lakes which differed in their trophic status, distribution of submerged macrophytes, and restoration history. PCR-denaturing gradient gel electrophoresis analyses demonstrated that the epiphytic compartment was inhabited by species belonging to cluster 3 of the Nitrosospira lineage and to the Nitrosomonas oligotropha lineage. Both the ammonia-oxidizing bacterial community compositions and the potential activities differed significantly between compartments. Interestingly, both the ammonia-oxidizing bacterial community composition and potential activity were influenced by the restoration status of the different lakes investigated.

Impact of acetochlor on ammonia-oxidizing bacteria in microcosm soils

Acetochlor is an increasingly used herbicide on corn in North China. Currently, the effect of acetochlor on soil ammonia-oxidizing bacteria (AOB) communities is not well documented. Here, we studied the diversity and community composition of AOB in soil amended with three concentrations of acetochlor (50, 150, 250 mg/kg) and the control (0 mg acetochlor/kg soil) in a microcosm experiment by PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and the phylogenetic analysis of excised DGGE bands. DGGE profiles showed that acetochlor had a stimulating effect on AOB at the early stage after acetochlor amended, and the order of intensity and duration is medium-acetochlor amended samples (AM) > low-acetochlor amended samples (AL) > high-acetochlor amended samples (AH). At the end of 60 d microcosm, acetochlor had a negative effect on the diversity of AOB. Cluster analysis of DGGE profiles showed that acetochlor had a greater effect on the community structure of AOB on day 60 than on day 1. The phylogenetic analysis revealed that all the sequences of excised DGGE bands were closely related to members of the genus Nitrosospira and formed two separate subclusters designated as subcluster 1 and subcluster 2 affiliated respectively with clusters 3 and 4 in Nitrosospira as defined by Stephen. Some dominant AOB had a change from subcluster 2 to subcluster 1 with the incubation. The results showed that acetochlor had an effect on the AOB on a long-term basis and the chronic effect of acetochlor should be paid more attention in future.

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.

Community composition of ammonia-oxidizing bacteria in the rhizosphere of intercropped wheat (Triticum aestivum L.), maize (Zea mays L.), and faba bean (Vicia faba L.)

Cereal/cereal and cereal/legume intercropping systems are popular in the north, northwest, and southwest of China and often result in yield increases compared to monocropping. Rhizosphere interactions may play a significant role in the yield increases, particularly with respect to nutrient availability. The aim of this study was to investigate the effects of intercropping on N availability and community composition of ammonia-oxidizing bacteria in the rhizosphere of wheat, maize, and faba bean at different growth stages. Denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes was used to analyze the community composition of bacterial ammonia oxidizers belonging to β-proteobacteria. The results showed that intercropping with faba bean significantly increased nitrate concentrations in the rhizosphere of wheat and maize at the second sampling time (20 June) compared to monocropping or intercropping between maize and wheat. Intercropping significantly affected the community composition of ammonia-oxidizing bacteria in the rhizosphere compared to monocropping, and the effects were most pronounced in the maize/faba bean and wheat/maize intercropping systems when faba bean and wheat were at anthesis and maize was in seedling stage. In wheat/faba bean intercropping, the effects of intercropping on community composition of ammonia-oxidizing bacteria were less pronounced at the seedling stage of the two species but were significant at anthesis.