Role of amendments on N cycling in Mediterranean abandoned semiarid soils

Soils found in semiarid areas of the Mediterranean Basin are particularly prone to degradation due to adverse climatic conditions with annual rainfall <300 mm and high temperatures being responsible for the scant vegetal growth and the consequent lack of organic matter. A three-year field experiment was conducted to test the potential of two organic amendments (sludge and compost) to improve soil quality and plant growth in a semiarid degraded Mediterranean ecosystem. Since little is known about N dynamics in such assisted ecosystems, we investigated the effects of this practice on key processes of the global N cycle. Besides soil chemical and biological parameters and vegetation cover, we measured absolute and specific potential nitrification and denitrification rates and quantified the size of the ammonia oxidising and denitrifying bacterial populations via quantitative PCR (amoA and nirS genes). At the end of the experiment soil fertility, microbial activity and plant growth had improved in treated plots. Amendments increased the amount of ammonia oxidisers and denitrifiers in soil, but the relative proportion of these groups varied in relation to the total microbial community, being higher in the case of ammonia oxidisers but not in the case of denitrifiers. As a consequence, significantly higher potential nitrification and denitrification rates were measured on a global basis in amended soils. Yet specific activities (potential rate/gene copy numbers) were lower for ammonia oxidisers in amended soils and for denitrifiers in sludge treated soils than those observed in control plots. Organic amendments influenced resource availability, the size and the activity patterns of microbial populations involved in long-term N dynamics. Therefore N cycling processes may play a key role to assist sustainable restoration practices in semiarid degraded areas.     

Response of ammonia oxidizing archaea and bacteria to changing water filled pore space

This study examined the effect of water filled pore space (WFPS) on gross N fluxes and community structure and abundance of ammonia oxidizing archaea and bacteria in a semi-arid soil. Different WFPS altered the community structure of both AOA and AOB. Ammonia oxidizer communities (for both archaea and bacteria) from ‘wet’ soils (95, 85 and 75% WFPS) and ‘dry’ soils (25, 45 and 55% WFPS) were distinctly different from one another. Additionally there was a significant relationship between community structure and gross rates of nitrification. There was also a significant relationship between WFPS and bacterial amoA abundance but not archaeal amoA abundance suggesting that bacterial ammonia oxidizers are more responsive to changes in soil water availability. These results are in agreement with other studies suggesting that both groups of ammonia oxidizers have distinct physiological characteristics and ecological niches with consequences for nitrification in response to WFPS. Overall findings from this study indicate that nitrification, both in terms of process rates and populations responsible for nitrification activity, is highly responsive to soil water availability.     

Responses of ammonia-oxidizing bacteria and archaea to nitrogen fertilization and precipitation increment in a typical temperate steppe in Inner Mongolia

As the first and rate-limiting step of nitrification, ammonia oxidation can be realized either by ammonia-oxidizing bacteria (AOB) or archaea (AOA). However, the key factors driving the abundance, community structure and activity of ammonia oxidizers are still unclear, and the relative importance of AOA and AOB in ammonia oxidation is unresolved. In the present study, we examined the effects of long-term (6 years) nitrogen (N) addition and simulated precipitation increment on the abundance and community composition of AOA and AOB based on a field trial in a typical temperate steppe of northern China. We used combined approaches of quantitative PCR, terminal-restriction fragment length polymorphism (T-RFLP) and clone library analyses of amoA genes. The study objective was to determine (1) AOA and AOB diversity and activity in response to N addition and increased precipitation and (2) the relative contributions of AOA and AOB to soil ammonia oxidation in the typical temperate steppe. The results showed that the potential nitrification rate (PNR) increased with N addition, but decreased with increased precipitation. Both N addition and increased precipitation significantly increased AOB but not AOA abundance, and a significant correlation was only observed between PNR and AOB amoA gene copies. The T-RFLP analysis showed that both N and precipitation were key factors in shaping the composition of AOB, while AOA were only marginally influenced. Phylogenetic analysis indicated that all AOA clones fell within the soil and sediment lineage while all AOB clones fell within the Nitrosospira. The study suggested that AOA and AOB had distinct physiological characteristics and ecological niches. AOB were shown to be more sensitive to N and precipitation than AOA, and the ammonia oxidation process was therefore supposed to be mainly driven by AOB in this temperate steppe.     

Nitrapyrin decreased nitrification of nitrogen released from soil organic matter but not amoA gene abundance at high soil temperature

Water pulses have a significant impact on nitrogen (N) cycling, making management of N challenging in agricultural soils that are exposed to episodic rainfall. In hot, dry environments, wetting of dry soil during summer fallow causes a rapid flush of organic matter mineralisation and subsequent nitrification, which may lead to N loss via nitrous oxide emission and nitrate leaching. Here we examined the potential for the nitrification inhibitor nitrapyrin to decrease gross nitrification at elevated temperature in soils with contrasting soil organic matter contents, and the consequent effects on ammonia oxidiser populations. Soil was collected during summer fallow while dry (water content 0.01 g g⁻¹ soil) from a research site with two management treatments (tilled soil and tilled soil with long-term additional crop residues) by three field replicates. The field dry soil (0–10 cm) was wet with or without nitrapyrin, and incubated (20 or 40 °C) at either constant soil water content or allowed to dry (to simulate summer drying after a rainfall event). Gross N transformation rates and inorganic N pools sizes were determined on six occasions during the 14 day incubation. Bacterial and archaeal amoA gene abundance was determined on days 0, 1, 7 and 14. Nitrapyrin increased ammonium retention and decreased gross nitrification rates even with soil drying at 40 °C. Nitrification was likely driven by bacterial ammonia oxidisers, as the archaeal amoA gene was below detection in the surface soil layer. Bacterial ammonia oxidiser gene abundances were not affected by nitrapyrin, despite the decrease in nitrifier activity. Increased soil organic matter from long-term additional crop residues diminished the effectiveness of nitrapyrin. The present study highlights the potential for nitrapyrin to decrease nitrification and the risk of N loss due to mineralisation of soil organic matter under summer fallow conditions.     

Fluctuations in the abundance of ammonia oxidizers and the contents of inorganic nitrogen in solarized soil

Solarization makes a great impact on the abundance of ammonia oxidizers and nitrifying activity in soil. To elucidate fluctuations in the abundance of ammonia oxidizers and nitrification in solarized soil, copy numbers of amoA gene of ammonia-oxidizing bacteria (AOB) and archaea (AOA), viable number of ammonia oxidizers and inorganic nitrogen contents were investigated in greenhouse experiments. The copy number of amoA gene and the viable number of ammonia oxidizers were determined by the quantitative polymerase chain reaction and most probable number methods, respectively. Abundance of AOB based on the estimation of amoA gene copy numbers and viable counts of ammonia oxidizers was decreased by the solarization treatment and increased during the tomato (Solanum lycopersicum L.) cultivation period following the solarization. Effect of solarization on the copy number of amoA gene of AOA was less evident than that on AOB. The proportion of nitrate in inorganic nitrogen contents was declined by the solarization and increased during the tomato cultivation period following the solarization. Positive correlations were found between the proportion of nitrate in inorganic nitrogen content and the copy number of bacterial or archaeal amoA gene or the viable number of ammonia oxidizers; the copy number of bacterial amoA gene showed a strong correlation with the viable number of ammonia oxidizers. The present study revealed influences of solarization on the fluctuation in the abundance of ammonia oxidizers and dynamics of inorganic nitrogen contents in soil and the results indicate that the determination of amoA gene of AOB is possibly a quick and useful diagnostic technique for evaluating suppression and restoration of nitrification following solarization.     

两种农田土壤中的氨氧化细菌和氨氧化古菌对硝化抑制剂DCD和DMPP的响应-盆栽试验

Taking two important agricultural soils with different pH, brown soil （Hap-Udic Luvisol） and cinnamon soil （Hap-Ustic Luvisol）, from Northeast China, a pot culture experiment with spring maize （Zea mays L.） was conducted to study the dynamic changes in the abundance and diversity of soil ammonia-oxidizing bacteria （AOB） and ammonia-oxidizing archaea （AOA） populations during maize growth period in response to the additions of nitrification inhibitors dicyandiamide （DCD） and 3,4-dimethylpyrazole phosphate （DMPP） by the methods of real-time polymerase chain reaction （PCR） assay, PCR-denaturing gradient gel electrophoresis （DGGE）, and construction of clone library targeting the amoA gene. Four treatments were established, i.e., no urea （control）, urea, urea plus DCD, and urea plus DMPP. Both DCD and DMPP inhibited growth of AOB significantly, compared to applying urea alone. Soil bacterial amoA gene copies had a significant positive linear correlation with soil nitrate content, but soil archaeal amoA gene copies did not. In both soils, all AOB sequences fell within Nitrosospira or Nitrosospira-like groups, and all AOA sequences belonged to group 1.1b crenaxchaea. With the application of DCD or DMPP, community composition of AOB and AOA in the two soils had less change except that the AOB community composition in Hap-Udic Luvisol changed at the last two growth stages of maize under the application of DCD. AOB rather than AOA likely dominated soil ammonia oxidation in these two agricultural soils.     

Nitrification activity and putative ammonia-oxidizing archaea in acidic red soils

Purpose  

Acidic red soils account for 21% of land area in China and contain low ammonia concentration due to ionization to ammonium. The unusual high affinity for ammonia of marine Nitrosopumilus maritimus and acidophilic soil Nitrosotalea devanaterra has suggested that ammonia-oxidizing archaea (AOA) may have greater selective advantage over ammonia-oxidizing bacteria (AOB) in ammonia-limited environment because ammonia rather than ammonium is thought to be the actual substrate for oxidation. The aim of this study was to assess whether nitrification activity can be attributed to AOA and/or AOB by relating community structures of AOA and AOB to nitrification activity in acidic red soils in southern China.     

Effects of irrigation sources on ammonia-oxidizing bacterial communities in a managed turf-covered aridisol

Ammonia-oxidizing bacteria (AOB) perform the rate-limiting step of nitrification, a key process in the global nitrogen cycle. In this study, chemical factors controlling AOB activity, diversity, and composition in a turfgrass-covered aridisol irrigated with groundwater, Colorado River water, or reclaimed wastewater were examined. Activity of AOB contributed an average of 96% of potential nitrification activity in four soils examined, and this activity correlated positively with ammonium concentration and negatively with salinity of the irrigation water. AOB abundance, as determined by quantitative polymerase chain reaction, also correlated positively with ammonium concentration in the irrigation water but negatively with soil salinity. Characterization of AOB communities by denaturing gradient gel electrophoresis showed the presence in every soil of AOB taxa, most commonly found in high-ammonia environments. The soil with the fewest years of management had the least diverse AOB population, compared to the other three soils, and much lower specific nitrification activity. This soil was irrigated with highly saline Colorado River water, which likely exerted acute negative effects on the activity of AOB. In summary, this study revealed that, although AOB activity and growth responded positively to ammonium availability in irrigation water, the salinity of the water and soil had strong negative effects on these aspects of the AOB community.     

基于宏基因组学的酸性森林土壤氨氧化微生物群落特征研究

全球30%以上陆地面积是酸性土壤(pH5.5),而酸性土壤中氨氧化微生物群落特征研究是破译其硝化过程微生物学机理的基础。尤其随着完全硝化微生物(Complete ammonia oxidizer,comammox)的发现,亟需重新认知酸性土壤中氨氧化微生物类群。以酸性马尾松林为研究对象,综合利用荧光定量PCR(qPCR)、凝胶电泳半定量和宏基因组测序等技术研究土壤中氨氧化古菌(Ammonia-oxidizing archaea,AOA)、氨氧化细菌(Ammonia-oxidizing bacteria,AOB)和Comammox的相对丰度以及群落组成特征。研究发现AOA和AOB amoA基因丰度分别为2.61×106 copies·g~(-1)和1.45×106copies·g~(-1);而comammoxamoA基因qPCR结果存在显著的非特异性扩增,导致其丰度被高估,而经凝胶电泳半定量矫正后,约为(1.38～1.47)×106copies·g~(-1),该结果和土壤宏基因测序揭示的comammox相对丰度基本吻合。此外,宏基因组分析发现经典嗜酸group1.1a-associated仅占AOA总类群的12%,而group1.1b则占88%,尽管目前仍未有嗜酸group 1.1b AOA纯菌株的报道。AOB主要类群为Nitrosospira(约64%),而Nitrosomonas约占36%。Comammox主要类群为clade B(约64%),而clade A仅占36%且均隶属于clade A.1亚枝,这暗示clade B与已报道的嗜中性comammox clade A纯菌株有极大的生理代谢差异。总之,本研究提供了综合利用qPCR、半定量和宏基因组分析土壤氨氧化微生物群落的策略,并建议优化comammox的qPCR引物,同时本研究系统分析了酸性马尾松林土壤中氨氧化微生物的相对丰度和群落组成特征。     

Ammonia-oxidizing archaea are more important than ammonia-oxidizing bacteria in nitrification and NO3 −-N loss in acidic soil of sloped land

As part of a long-term sloped land use experiment established in 1995 at Taoyuan Agro-ecosystem Research Station (111°26′ E, 28°55′ N) in China, soil samples were collected from three land use types, including cropland (CL), natural forest, and tea plantation. Quantitative polymerase chain reaction and terminal restriction fragment length polymorphism were used to determine the abundance and community composition of amoA-containing bacteria (AOB) and archaea (AOA). The results indicate that land use type induced significant changes in soil potential nitrification rate and community composition, diversity, and abundance of AOB and AOA. Both AOB and AOA community compositions were generally similar between upper and lower slope positions (UP and LP), except within CL. The LP soils had significantly (p?<?0.05) higher diversity and abundance of both AOB and AOA than in the UP. Potential nitrification rate was significantly correlated (p?<?0.05) with diversity and abundance of AOA, but not with AOB. Among land use types, the NO₃ ^? and amoA-containing AOA runoff loss was greatest in CL. Nitrate-N runoff loss was significantly correlated (p?<?0.05) with the loss of AOA amoA copies in the runoff water. Furthermore, relationships between NO₃ ^?-N runoff loss and abundance of AOA but not of AOB at both slope positions were significantly correlated (p?<?0.05). These findings suggest that AOA are more important than AOB in nitrification and NO₃ ^?-N runoff loss in acidic soils across sloped land use types.     

pH regulates key players of nitrification in paddy soils

Increasing lines of evidence have suggested the functional importance of ammonia-oxidizing archaea (AOA) rather than bacteria (AOB) for nitrification in upland soils with low pH. However, it remains unclear whether niche specialization of AOA and AOB occurs in rice paddy wetlands constrained by oxygen availability. Using DNA-based stable isotope probing, we conclude that AOA dominated nitrification activity in acidic paddy soils (pH 5.6) while AOB dominated in alkaline soils (pH 8.2). Nitrification activity was stimulated by urea fertilization and accompanied by a significant increase of AOA in acid soils and AOB in alkaline soils. DNA-based stable isotope probing indicated significant assimilation of ¹³CO₂ for AOA only in acidic paddy soil, while AOB was the solely responsible for ammonia oxidation in the alkaline paddy soil. Phylogenetic analysis further indicated that AOA members within the soil group 1.1b lineage dominated nitrification in acid soils. Ammonia oxidation in the alkaline soil was catalyzed by Nitrosospira cluster 3-like AOB, suggesting that the physiological diversity of AOA is more complicated than previously thought, and soil pH plays important roles in shaping the community structures of ammonia oxidizers in paddy field.     

Nitrapyrin affects the abundance of ammonia oxidizers rather than community structure in a yellow clay paddy soil

Purpose

Yellow clay paddy soil (Oxisols) is a low-yield soil with low nitrogen use efficiency (NUE) in southern China. The nitrification inhibitor nitrapyrin (2-chloro-6- (tricholoromethyl)-pyridine, CP) has been applied to improve NUE and reduce environmental pollution in paddy soil. However, the effects of nitrapyrin combined with nitrogen fertilizers on ammonia oxidizers in yellow clay paddy soil have not been examined.

Materials and methods

A randomized complete block design was set with three treatments: (1) without nitrogen fertilizer (CK), (2) common prilled urea (PU), and (3) prilled urea with nitrapyrin (NPU). Soil samples were collected from three treatments where CK, PU, and NPU had been repeatedly applied over 5 years. Soil samples were analyzed by quantitative PCR and 454 high-throughput pyrosequencing of the amoA gene to investigate the influence of nitrapyrin combined with nitrogen on the abundance and community structure of ammonia oxidizers in yellow clay paddy soil.

Results and discussion

The potential nitrification rate (PNR) of the soil was significantly correlated with the abundances of both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). Application of urea significantly stimulated AOA and AOB growth, whereas nitrapyrin exhibited inhibitory effects on AOA. Phylogenetic analysis showed that the most dominant operational taxonomic units (OTUs) of AOA and AOB were affiliated with the Nitrosotalea cluster and Nitrosospira cluster 12, respectively. AOA and AOB community structures were not altered by urea and nitrapyrin application.

Conclusions

Nitrogen fertilization stimulated nitrification and increased the population sizes of AOA and AOB. Nitrapyrin affected the abundance, but not community structure of ammonia oxidizers in yellow clay soil. Our results suggested that nitrapyrin improving NUE and inhibiting PNR was attributable to the inhibition of AOA growth.

     

Do climate warming and plant species richness affect potential nitrification, basal respiration and ammonia-oxidizing bacteria in experimental grasslands?

Ammonia-oxidizing bacteria (AOB) are key organisms in the N cycle, as they control the first, rate-limiting step of the nitrification process. The question whether current environmental disturbances, such as climate warming and plant diversity losses, select for a particular community structure of AOB and/or influence their activity remains open. The purpose of this research was to study the impact of a 3 °C warming and of plant species richness (S) on microbial activity and diversity in synthesized grasslands, with emphasis on the nitrification process and on the diversity (community structure and richness) of ammonia-oxidizing bacteria (AOB). We measured soil chemical characteristics, basal respiration, potential nitrification and AOB diversity in soils under increasing plant species richness (S = 1, S = 3, S = 9) at ambient and (ambient +3 °C) temperature. Species were drawn from a 9-species pool, belonging to three functional groups: forbs, legumes and grasses. Mixtures comprised species from each of the three functional groups. Warming did not affect AOB diversity and increased potential nitrification at S = 3 only. Under warmed conditions, higher plant species richness resulted in increased potential nitrification rates. AOB richness increased with plant species richness. AOB community structure of monocultures under legumes differed from those under forbs and grasses. Clustering analysis revealed that AOB community structure under legume monocultures and mixtures of three and nine species grouped together. These results indicate that functional group identity rather than plant species richness influenced AOB community structure, especially through the presence of legumes. No clear relationship emerged between AOB richness and potential nitrification whatever plant species richness and temperature treatment. Our findings show a link between aboveground and belowground diversity, namely plant species richness, AOB richness and community structure. AOB richness was not related to soil processes, supporting the idea that increased diversity does not necessarily lead to increased rates of ecosystem processes.     

Different roles of rhizosphere effect and long-term fertilization in the activity and community structure of ammonia oxidizers in a calcareous fluvo-aquic soil

Ammonia oxidation is a critical step in the soil nitrogen (N) cycle and can be affected by the application of mineral fertilizers or organic manure. However, little is known about the rhizosphere effect on the function and structure of ammonia-oxidizing bacterial (AOB) and archaeal (AOA) communities, the most important organisms responsible for ammonia oxidation in agricultural ecosystems. Here, the potential nitrification activity (PNA), population size and composition of AOB and AOA communities in both the rhizosphere and bulk soil from a long-term (31-year) fertilizer field experiment conducted during two seasons (wheat and maize) were investigated using the shaken slurry method, quantitative real-time polymerase chain reaction and denaturing gradient gel electrophoresis. N fertilization greatly enhanced PNA and AOB abundance, while manure application increased AOA abundance. The community structure of AOB exhibited more obvious shifts than that of AOA after long-term fertilization, resulting in more abundant AOB phylotypes similar to Nitrosospira clusters 3 and 4 in the N-fertilized treatments. Moreover, PNA was closely correlated with the abundance and community structure of AOB rather than that of AOA among soils during both seasons, indicating that AOB play an active role in ammonia oxidation. Conversely, the PNA and population sizes of AOB and AOA were typically higher in the rhizosphere than the bulk soil, implying a significant rhizosphere effect on ammonia oxidation. Cluster and redundancy analyses further showed that this rhizosphere effect played a more important role in shaping AOA community structure than long-term fertilization. Overall, the results indicate that AOB rather than AOA functionally dominate ammonia oxidation in the calcareous fluvo-aquic soil, and that rhizosphere effect and fertilization regime play different roles in the activity and community structures of AOB and AOA.     

The relative contribution of nitrifiers to autotrophic nitrification across a pH-gradient in a vegetable cropped soil

Purpose

Microbial nitrification plays an important role in nitrogen cycling in ecosystems. Nitrification is performed by ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), and nitrite-oxidizing bacteria (NOB) including complete ammonia oxidizers. However, the relative importance of nitrifiers in autotrophic nitrification in relation to soil pH is still unclear.

Materials and methods

Combining DNA-based stable isotope probing (SIP) and molecular biological techniques, we investigated the abundance, structure, and activity of AOA, AOB, and NOB along a pH-gradient (3.97–7.04) in a vegetable cropped soil.

Results and discussion

We found that AOA abundance outnumbered AOB abundance and had a significantly negative relationship with soil pH. The abundances of NOB Nitrospira 16S rRNA, nxrB gene, and Nitrobacter nxrA gene were affected by soil pH. Incubation of soil with ¹³CO₂ and DNA-SIP analysis demonstrated that significant ¹³CO₂ assimilation by AOA rather than by AOB occurred in the acidic soils, whereas the labeled ¹³C level of AOA was much less in the neutral soil than in the acidic soils. There was no evidence of ¹³CO₂ assimilation by NOB except for Nitrobacter with NxrB gene at pH 3.97. Phylogenetic analysis of AOA amoA gene in the ¹³C- and ¹²C-labeled treatments showed that the active AOA mainly belonged to Nitrososphaera in the acidic soils.

Conclusions

These results suggested that the main performer of nitrification was AOA in the acidic soils, but both AOA and AOB participated in nitrification in the neutral soil with low nitrification activity. NOB Nitrospira and Nitrobacter did not grow in the soils with pH 4.82–7.04 and other populations of NOB were probably involved in nitrite oxidation in the vegetable cropped soil.

     

Contrasting response of nitrification capacity in three agricultural soils to N addition during short-term incubation

Purpose

Human disturbance is a major culprit driving imbalances in the biological transformation of nitrogen from the nonreactive to the reactive pool and is therefore one of the greatest concerns for nitrogen (N) cycling. The objective of this study was to compare potential nitrification rates and the abundance of ammonia oxidizers responsible for nitrification, with the amendment of external N in different agricultural soils.

Materials and methods

Three typical Chinese agricultural soils, QiYang (QY) acid soil, ShenYang (SY) neutral soil, and FengQiu (FQ) alkaline soil, were amended with 0, 20, 150, and 300 μg NH₄ ⁺-N g^?1 soil and incubated for 40 days. The abundance of ammonia oxidizing bacteria (AOB) and archaea (AOA) at the end of incubation in the soil microcosms was determined using the real-time PCR.

Results and discussion

There was a significant decrease in ammonium concentration in the QY soil from the highest to the lowest N-loading treatments, while no significant difference in ammonium concentrations was detected among the different N-loading treatments for the SY and FQ soils. A significantly higher potential nitrification rate (PNR) was observed in the FQ soil while lowest PNR was found in the QY soil. Quantitative PCR analysis of AOB amoA genes demonstrated that AOB abundance was significantly higher in the high N-loading treatments than in the control for the QY soil only, while no significant difference among treatments in the SY and FQ soils. A significant positive correlation between PNR and AOB amoA abundance, however, was found for the SY and FQ soils, but not for the QY soil. Little difference in AOA amoA abundance between different N-loading treatments was observed for all the soils.

Conclusions

This study suggested that ammonia oxidation capacity in the FQ and SY soils was higher than those in the QY soil with the addition of ammonium fertilizer for a short-term. These findings indicated that understanding the differential responses of biological nitrification to varying input levels of ammonium fertilizer is important for maximizing N use efficiency and thereby improving agricultural fertilization management.     

Impact of irrigation water quality on soil nitrifying and total bacterial communities

Disturbance induced by two contrasting irrigation regimes (groundwater versus urban wastewater) was evaluated on a sandy agricultural soil through chemical and microbial analyses. Contrary to wastewater, groundwater displayed very high nitrate contents but small amounts of ammonium and organic matter. Despite these strong compositional shifts, soil organic carbon and nitrogen, nitrate and ammonium contents were not significantly different in both types of irrigated plot. Moreover, neither microbial biomass nor its activity, determined as fluorescein diacetate hydrolysis activity, was influenced by irrigation regimes. Bacterial community structure, assessed by denaturing gradient gel electrophoresis (DGGE) of 16S ribosomal DNA fragments, was also weakly impacted as molecular fingerprints shared an overall similarity of 85%. Ammonia-oxidizing bacterial community (AOB) was monitored by DGGE of the functional molecular marker amoA gene (alpha subunit of the ammonia monooxygenase). Surprisingly, no amoA signals were obtained from plots irrigated with groundwater, whereas signal intensities were high in all plots under wastewater. Among the last, compositional shifts of the AOB community were weak. Overall, impact of irrigation water quality on soil chemistry could not be evidenced, whereas effects were low on the total bacterial compartment but marked on the AOB community.     

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.     

Net nitrification rate and presence of Nitrosospira cluster 2 in acid coniferous forest soils appear to be tree species specific

The impact of four coniferous tree species and their corresponding soil factors on N transformation rates and presence of ammonia-oxidising bacteria (AOB) was studied in an acid pine forest soil (Appelscha, The Netherlands). Pine soil had a relatively low net nitrification rate, while spruce, fir and larch soils showed high net nitrification rates. 16S rRNA and amoA sequences were only found in soils with high nitrification rates and belonged solely to Nitrosospira cluster 2. We conclude that tree species, possibly through their effects on soil C/N ratios, determines the presence of Nitrosospira cluster 2. Whenever AOB are present, however, the AOB community composition appears to be similar.     

Management practices have a major impact on nitrifier and denitrifier communities in a semiarid grassland ecosystem

Purpose

Nitrification and denitrification, two of the key nitrogen (N) transformation processes in the soil, are carried out by a diverse range of microorganisms and catalyzed by a series of enzymes. Different management practices, such as continuous grazing, mowing, and periodic fencing off from grazing, dramatically influenced grassland ecosystems. This study aimed to examine the effects of management practices on the abundance and community structure of nitrifier and denitrifier communities in grassland ecosystems.

Materials and methods

Soil samples were collected from a semiarid grassland ecosystem in Xilingol region, Inner Mongolia, where long-term management practices including free-grazing, different periods of enclosure from grazing, and different frequencies of mowing were conducted. Real-time quantitative polymerase chain reaction (Q-PCR), denaturing gradient gel electrophoresis (DGGE), sequencing, and phylogenetic analysis were applied to estimate the abundance and composition of amoA, nirS, nirK, and nosZ genes.

Results and discussion

The ammonia-oxidizing archaea (AOA) amoA copies were in the range 5.99?×?10⁸ to 8.60?×?10⁸, while those of ammonia-oxidizing bacteria (AOB) varied from 3.02?×?10⁷ to 4.61?×?10⁷. The abundance of AOA was substantially higher in the light grazing treatment (LG) than in the mowing treatments. The quantity and intensity of DGGE bands of AOA varied with pasture management. In stark contrast, AOB population abundance and community structure remained largely unchanged in all the soils irrespective of the management practices. All these results suggested that ammonia oxidizers were dominated by AOA. The higher gene abundance and greater intensity of DGGE bands of nirS and nosZ under the enclosure treatments would suggest greater stimulated denitrification. The ratio of nosZ/(nirS?+?nirK) was higher in mowing treatments than in the free-grazing and enclosure treatments, possibly leading to more complete denitrification. Correlation analysis indicated that soil moisture and inorganic nitrogen content were the two main soil environmental variables that influence the community structure of nitrifiers and denitrifiers.

Conclusions

In this semiarid neutral to alkaline grassland ecosystem under low temperature conditions, AOA mainly affiliated with Nitrososphaera dominated nitrification. These results clearly demonstrate that grassland management practices can have a major impact on nitrifier and denitrifier communities in this semiarid grassland ecosystem, under low temperature conditions.