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 methodsCombining 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 discussionWe 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.
ConclusionsThese 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.
相似文献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 methodsA 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 discussionThe 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.
ConclusionsNitrogen 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.
相似文献Nitrification and denitrification in the N cycle are affected by various ammonia oxidizers and denitrifying microbes in intensive vegetable cultivation soils, but our current understanding of the effect these microbes have on N2O emissions is limited. The nitrification inhibitor, 3,4-dimethylpyrazole phosphate (DMPP), acts by slowing nitrification and is used to improve fertilizer use efficiency and reduce N losses from agricultural systems; however, its effects on nitrifier and denitrifier activities in intensive vegetable cultivation soils are unknown.
Materials and methodsIn this study, we measured the impacts of DMPP on N2O emissions, ammonia oxidizers, and denitrifying microbes in two intensive vegetable cultivation soils: one that had been cultivated for a short term (1 year) and one that had been cultivated over a longer term (29 years). The quantitative PCR technique was used in this study. Three treatments, including control (no fertilizer), urea alone, and urea with DMPP, were included for each soil. The application rates of urea and DMPP were 1800 kg ha?1 and 0.5% of the urea-N application rate.
Results and discussionThe application of N significantly increased N2O emissions in both soils. The abundance of ammonia-oxidizing bacteria (AOB) increased significantly with high rate of N fertilizer application in both soils. Conversely, there was no change in the growth rate of ammonia-oxidizing archaea (AOA) in response to the applied urea despite the presence of larger numbers of AOA in these soils. This suggests AOB may play a greater role than AOA in the nitrification process, and N2O emission in intensive vegetable cultivation soils. The application of DMPP significantly reduced soil NO3?-N content and N2O emission, and delayed ammonia oxidation. It greatly reduced AOB abundance, but not AOA abundance. Moreover, the presence of DMPP was correlated with a significant decrease in the abundance of nitrite reductase (nirS and nirK) genes.
ConclusionsLong-term intensive vegetable cultivation with heavy N fertilization altered AOB and nirS abundance. In vegetable cultivation soils with high N levels, DMPP can be effective in mitigating N2O emissions by directly inhibiting both ammonia oxidizing and denitrifying microbes.
相似文献Complete ammonia oxidizers (comammox), which convert ammonia to nitrate through nitrite, are a newly discovered nitrifying group. In recent years, comammox Nitrospira have been discovered in various natural and engineered ecosystems. However, little is known about the distribution dynamics of comammox Nitrospira in estuary tidal flat sediments.
Materials and methodsChongming eastern tidal flat, the largest tidal flat in the Yangtze River Estuary, was selected as the research area. Through a combination of molecular biology assays, phylogenetic analysis based on functional gene sequences and statistical correlation with physicochemical properties, we determined the distribution and diversity of comammox Nitrospira in Chongming eastern tidal flat and analyzed the potential influencing environmental factors.
Results and discussionThe results indicated comammox Nitrospira were widely distributed in Chongming eastern tidal flat, while with lower abundance than the ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA). Clade A1 comammox Nitrospira showed adaptation to relatively high salinity, and was more distributed in middle and low tidal flats, while clade A2 and clade A3 were mostly distributed in high tidal flats with low salinity. The abundance and community structure of comammox Nitrospira were mainly affected by salinity, ammonia concentration, and temperature.
ConclusionThis study showed the general existence of comammox Nitrospira in Chongming eastern intertidal sediments and indicated that the differences of tidal locations, which lead to a gradient in the physicochemical properties of the sediments, in turn affects the spatial distribution of comammox Nitrospira in estuary tidal flats.
相似文献Purpose
Nitrification, the microbial oxidation of ammonia to nitrate via nitrite, is a pivotal component of the biogeochemical nitrogen cycle. Nitrification was conventionally assumed as a two-step process in which ammonia oxidation was thought to be catalyzed by ammonia-oxidizing archaea (AOA) and bacteria (AOB), as well as nitrite oxidation by nitrite-oxidizing bacteria (NOB). This long-held assumption of labour division between the two functional groups, however, was challenged by the recent unexpected discovery of complete ammonia oxidizers within the Nitrospira genus that are capable of converting ammonia to nitrate in a single organism (comammox). This breakthrough raised fundamental questions on the niche specialization and differentiation of comammox organisms with other canonical nitrifying prokaryotes in terrestrial ecosystems.Materials and methods
This article provides an overview of the recent insights into the genomic analysis, physiological characterization and environmental investigation of the comammox organisms, which have dramatically changed our perspective on the aerobic nitrification process. By using quantitative PCR analysis, we also compared the abundances of comammox Nitrospira clade A and clade B, AOA, AOB and NOB in 300 forest soil samples from China spanning a wide range of soil pH.Results and discussion
Comammox Nitrospira are environmentally widespread and numerically abundant in natural and engineered habitats. Physiological data, including ammonia oxidation kinetics and metabolic versatility, and comparative genomic analysis revealed that comammox organisms might functionally outcompete other canonical nitrifiers under highly oligotrophic conditions. These findings highlight the necessity in future studies to re-evaluate the niche differentiation between ammonia oxidizers and their relative contribution to nitrification in various terrestrial ecosystems by including comammox Nitrospira in such comparisons.Conclusions
The discovery of comammox and their broad environmental distribution added a new dimension to our knowledge of the biochemistry and physiology of nitrification and has far-reaching implications for refined strategies to manipulate nitrification in terrestrial ecosystems and to maximize agricultural productivity and sustainability.Paddy fields are an important source of nitrous oxide (N2O) emission. The application of biochar or the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) to paddy soils have been proposed as technologies to mitigate N2O emissions, but their mechanisms remain poorly understood.
MethodsAn experiment was undertaken to study the combined and individual effects of biochar and DMPP on N2O emission from a paddy field. Changes in soil microbial community composition were investigated. Four fertilized treatments were established as follows: fertilizer only, biochar, DMPP, and biochar combined with DMPP; along with an unfertilized control.
ResultsThe application of biochar and/or DMPP decreased N2O emission by 18.9–39.6% compared with fertilizer only. The combination of biochar and DMPP exhibited higher efficiency at suppressing N2O emission than biochar alone but not as effective as DMPP alone. Biochar promoted the growth of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB), while DMPP suppressed AOB and increased AOA. Applying biochar with DMPP reduced the impact of DMPP on AOB. The nirS-/nirK- denitrifiers were decreased and nosZ-N2O reducers were increased by DMPP and the combination of DMPP and biochar. The abundance of the nirK gene was increased by biochar at the elongation and heading stages of rice development. Compared with fertilizer only, the application of biochar and/or DMPP promoted the abundance of nosZ genes.
ConclusionThese results suggest that applying biochar and/or DMPP to rice paddy fields is a promising strategy to reduce N2O emissions by regulating the dynamics of ammonia oxidizers and N2O reducers.
相似文献Ammonia oxidation is the limiting step in soil nitrification and critical in the global nitrogen cycle. The discovery of ammonia-oxidizing archaea (AOA) has improved our knowledge of microbial mechanisms for ammonia oxidation in complex soil environments. However, the relative contributions of AOA and ammonia-oxidizing bacteria (AOB) to ammonia oxidation remain unclear.
Materials and methodsIn this study, through large geographical scale sampling in China, totally nine samples representing various types of arable land soils were selected for analyzing the ammonia oxidation activity. The AOA and AOB activities were separately determined by using the dicyandiamide and 1-octyne inhibition method. High-throughput pyrosequencing and DNA stable-isotope probing (DNA-SIP) analysis were applied to investigate the distribution and activity of Candidatus Nitrosocosmicus franklandus in the arable land soils.
Results and discussionIn this study, AOA abundance (3.2?×?107–3.4?×?109 copies g?1) and activity (0.01–1.33 mg N kg?1 dry soil day?1) were evaluated for nine selected arable land soils and accounted for 4–100% of ammonia oxidation. By separately determining AOA and AOB rates, we observed that archaeal ammonia oxidation dominated the ammonia oxidation process in six soils, revealing a considerable contribution of AOA in ammonia oxidation in arable land soils. Based on high-throughput pyrosequencing analysis, the AOA species Ca. N. franklandus with relatively low abundance (0.6–13.5% in AOA) was ubiquitously distributed in all the tested samples. Moreover, according to the DNA-SIP analysis for Urumqi sample, the high activity and efficiency of Ca. N. franklandus in using CO2 suggests that this species plays an important role in archaeal ammonia oxidation in arable land soils.
ConclusionsThrough determining the AOA activity and analyzing the potential predominant functional AOA species, this study greatly improves our understanding of ammonia oxidation in arable land soils.
相似文献