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.     

Soil pH and plant diversity drive co-occurrence patterns of ammonia and nitrite oxidizer in soils from forest ecosystems

In this study, we investigated how co-occurrence patters of ammonia and nitrite oxidizers, which drive autotrophic nitrification, are influenced by tree species composition as well as soil pH in different forest soils. We expected that a decline of ammonia oxidizers in coniferous forests, as a result of excreted nitrification inhibitors and at acidic sites with low availability of ammonia, would reduce the abundance of nitrite-oxidizing bacteria (NOB). To detect shifts in co-occurrence patterns, the abundance of key players was measured at 50 forest plots with coniferous respectively deciduous vegetation and different soil pH levels in the region Schwäbische Alb (Germany). We found ammonia-oxidizing archaea (AOA) and Nitrospira-like NOB (NS) to be dominating in numbers over their counterparts across all forest types. AOA co-occurred mostly with NS, while bacterial ammonia oxidizers (AOB) were correlated with Nitrobacter-like NOB (NB). Co-occurrence patterns changed from tight significant relationships of all ammonia and nitrite oxidizers in deciduous forests to a significant relationship of AOB and NB in coniferous forests, where AOA abundance was reduced. Surprisingly, no co-occurrence structures between ammonia and nitrite oxidizers could be determined at acidic sites, although abundances were correlated to the respective nitrogen pools. This raises the question whether interactions with heterotrophic nitrifiers may occur, which needs to be addressed in future studies.     

The short-term effects of nitrification inhibitors on the abundance and expression of ammonia and nitrite oxidizers in a long-term field experiment comparing land management

Microcosms were set up to evaluate the effect of nitrification inhibitors (DCD, c-PTiO, and NaClO₃) on the abundance and expression of ammonia-oxidizing bacteria (AOB) and archaea (AOA), as well as the nitrite-oxidizing bacteria (NOB) Nitrospira and Nitrobacter. Both DCD and NaClO₃ inhibited the net nitrification rate, while c-PTiO had no significant effects, and NaClO₃ had a much greater inhibitory effect (> 60%) in all soils than DCD. No significant changes in total microbial abundance were observed with DCD and NaClO₃. DCD limited only the growth of AOB; however, NaClO₃ inhibited growth of both AOA and Nitrospira-NOB with no significant effects on AOB and Nitrobacter-NOB. Probably NaClO₃ inhibited both ammonia oxidation and nitrite oxidation. This is the first report to reveal the inhibitory effects of NaClO₃ on a specific nitrification process, helping to clarify the ecological niche of nitrifiers and the potential of nitrification inhibitors applied to soil.     

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 (N_CK), 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 N_CK 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.     

High contribution of ammonia-oxidizing archaea (AOA) to ammonia oxidation related to a potential active AOA species in various arable land soils

Purpose

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 methods

In 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 discussion

In 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.

Conclusions

Through 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.

     

Evidence for niche differentiation of nitrifying communities in grassland soils after 44 years of different field fertilization scenarios

Long-term nitrogen(N)fertilization imposes strong selection on nitrifying communities in agricultural soil,but how a progressively changing niche affects potentially active nitrifiers in the field remains poorly understood.Using a 44-year grassland fertilization experiment,we investigated community shifts of active nitrifiers by DNA-based stable isotope probing(SIP)of field soils that received no fertilization(CK),high levels of organic cattle manure(HC),and chemical N fertilization(CF).Incubation of DNA-SIP microcosms showed significant nitrification activities in CF and HC soils,whereas no activity occurred in CK soils.The 44 years of inorganic N fertilization selected only 13C-ammonia-oxidizing bacteria(AOB),whereas cattle slurry applications created a niche in which both ammonia-oxidizing archaea(AOA)and AOB could be actively13C-labeled.Phylogenetic analysis indicated that Nitrosospira sp.62-like AOB dominated inorganically fertilized CF soils,while Nitrosospira sp.41-like AOB were abundant in organically fertilized HC soils.The 13C-AOA in HC soils were affiliated with the 29i4 lineage.The 13C-nitrite-oxidizing bacteria(NOB)were dominated by both Nitrospira-and Nitrobacter-like communities in CF soils,and the latter was overwhelmingly abundant in HC soils.The 13C-labeled nitrifying communities in SIP microcosms of CF and HC soils were largely similar to those predominant under field conditions.These results provide direct evidence for a strong selection of distinctly active nitrifiers after 44 years of different fertilization regimes in the field.Our findings imply that niche differentiation of nitrifying communities could be assessed as a net result of microbial adaption over 44 years to inorganic and organic N fertilization in the field,where distinct nitrifiers have been shaped by intensified anthropogenic N input.     

Growth of comammox Nitrospira is inhibited by nitrification inhibitors in agricultural soils

Purpose

The discovery of comammox Nitrospira being capable of complete oxidising ammonia to nitrate radically challenged the conventional concept of two-step nitrification. However, the response of comammox Nitrospira to nitrification inhibitors (NIs) and their role in soil nitrification remain largely unknown, which has hindered our ability to predict the efficiency of NIs in agroecosystems.

Materials and methods

We evaluated the effect of four NIs, 2-chloro-6-(trichloromethyl) pyridine (nitrapyrin), 3,4-dimethylpyrazole phosphate (DMPP), allylthiourea (ATU) and dicyandiamide (DCD) on the growth of comammox Nitrospira, ammonia-oxidising archaea (AOA) and ammonia-oxidising bacteria (AOB) in two pasture and arable soils.

Results and discussion

The amendment of nitrogen fertiliser significantly increased soil nitrate concentrations over time, indicating a sustaining nitrification activity in both soils. The addition of all the four NIs effectively reduced the production of nitrate in both soils, but to varying degrees during incubation. The abundances of comammox Nitrospira clade A were significantly increased by addition of nitrogen fertilisers and significantly impeded by the four NIs in the pasture soil, but their abundances were only remarkably hindered by nitrapyrin in the arable soil. All the four NIs obviously inhibited the AOB abundances in both soils. Except for DMPP, the other three NIs effectively suppressed the AOA abundances in both soils.

Conclusions

We provided new evidence that growth of comammox Nitrospira clade A can be stimulated by nitrogen fertilisers and inhibited by various nitrification inhibitors, suggesting their potential role in nitrification of agricultural soils.

     

Effects of 3,4-dimethylpyrazole phosphate (DMPP) on the abundance of ammonia oxidizers and denitrifiers in two different intensive vegetable cultivation soils

Purpose

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 methods

In 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 discussion

The 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.

Conclusions

Long-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.

     

亚热带松林中强酸性土壤自养和异养硝化作用的研究

The occurrence of nitrification in some acidic forest soils is still a subject of debate. Identification of main nitrification pathways in acidic forest soils is still largely unknown. Acidic yellow soil (Oxisol) samples were selected to test whether nitrification can occur or not in acidic subtropical pine forest ecosystems. Relative contributions of autotrophs and heterotrophs to nitrification were studied by adding selective nitrification inhibitor nitrapyrin. Soil NH₄⁺-N concentrations decreased, but NO₃^--N concentrations increased significantly for the no-nitrapyrin control during the first week of incubation, indicating that nitrification did occur in the acidic subtropical soil. The calculated net nitrification rate was 0.49 mg N kg^-1 d^-1 for the no-nitrapyrin control during the first week of incubation. Nitrapyrin amendment resulted in a significant reduction of NO₃^--N concentration. Autotrophic nitrification rate averaged 0.28 mg N kg^-1 d^-1 and the heterotrophic nitrification rate was 0.21 mg N kg^-1 d^-1 in the first week. Ammonia-oxidizing bacteria (AOB) abundance increased slightly during incubation, but nitrapyrin amendment significantly decreased AOB amoA gene copy numbers by about 80%. However, the ammonia-oxidizing archaea (AOA) abundance showed significant increases only in the last 2 weeks of incubation and it was also decreased by nitrapyrin amendment. Our results indicated that nitrification did occur in the present acidic subtropical pine forest soil, and autotrophic nitrification was the main nitrification pathway. Both AOA and AOB were the active biotic agents responsible for autotrophic nitrification in the acidic subtropical pine forest soil.     

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.     

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。     

亚硝化球菌属(Nitrososphaera)可能是酸性土壤硝化作用的重要驱动者

选择初始pH相近的两个酸性土壤(JX-3和JX-7)样品进行培养试验,探讨了氨氧化古菌(ammonia-oxidizing archaea,AOA)和氨氧化细菌(ammonia-oxidizing bacteria,AOB)在酸性土壤硝化过程中所发挥的作用。结果显示,经过50 d的培养,JX-7样品硝化速率显著高于JX-3,且明显降低土壤pH。培养后,两个土壤样品AOB丰度均增加,但样品间没有显著差异;JX-7土壤AOA丰度显著增加,而JX-3无显著变化。两个土壤样品AOA群落组成本身存在分异,但对于同一样品培养前后均无显著分异;AOB群落组成在两土壤间没有分异,但培养前后分别有分异。培养后,JX-7样品中AOA优势属Nitrososphaera和某些未知微生物的个别OTUs绝对丰度显著增加,而两样品AOB中Nitrosospira属的一些OTUs的绝对丰度均显著增加。因此,所研究的酸性土壤样品中AOA是硝化作用的主要贡献者,而且AOA主要通过提高Nitrososphaera属中个别OTUs的丰度,而不是整个群落来调控硝化作用。     

Contributions of ammonia-oxidizing archaea and bacteria to nitrification in Oregon forest soils

Ammonia oxidation, the first step of nitrification, is mediated by both ammonia-oxidizing archaea (AOA) and bacteria (AOB); however, the relative contributions of AOA and AOB to soil nitrification are not well understood. In this study we used 1-octyne to discriminate between AOA- and AOB-supported nitrification determined both in soil-water slurries and in unsaturated whole soil at field moisture. Soils were collected from stands of red alder (Alnus rubra Bong.) and Douglas-fir (Pseudotsuga menziesii Mirb. Franco) at three sites (Cascade Head, the H.J. Andrews, and McDonald Forest) on acidic soils (pH 3.9–5.7) in Oregon, USA. The abundances of AOA and AOB were measured using quantitative PCR by targeting the amoA gene, which encodes subunit A of ammonia monooxygenase. Total and AOA-specific (octyne-resistant) nitrification activities in soil slurries were significantly higher at Cascade Head (the most acidic soils, pH < 5) than at either the H.J. Andrews or McDonald Forest, and greater in red alder compared with Douglas-fir soils. The fraction of octyne-resistant nitrification varied among sites (21–74%) and was highest at Cascade Head than at the other two locations. Net nitrification rates of whole soil without NH₄⁺ amendment ranged from 0.4 to 3.3 mg N kg⁻¹ soil d⁻¹. Overall, net nitrification rates of whole soil were stimulated 2- to 8-fold by addition of 140 mg NH₄⁺-N kg⁻¹ soil; this was significant for red alder at Cascade Head and the H.J. Andrews. Red alder at Cascade Head was unique in that the majority of NH₄⁺-stimulated nitrifying activity was octyne-resistant (73%). At all other sites, NH₄⁺-stimulated nitrification was octyne-sensitive (68–90%). The octyne-sensitive activity—presumably AOB—was affected more by soil pH whereas the octyne-resistant (AOA) activity was more strongly related to N availability.     

土地利用方式对万木林土壤氨氧化微生物丰度的影响

以我国亚热带地区典型花岗岩发育酸性红壤为研究对象,选取福建建瓯万木林自然保护区封禁保护下5种自然植被和1种人工种植植被土壤,采用荧光实时定量PCR(Real-time PCR)技术测定了土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)的群落丰度,采用15N稳定同位素成对标记和数值模型相结合的方法测定了土壤初级硝化速率。结果显示,长期封禁保护下的自然植被土壤pH低,土壤AOB数量偏低。人为种植和管理显著提高了土壤pH,促进了AOB的生长,其丰度比自然条件下提高了2个数量级,土壤初级硝化速率也显著提高,并与AOB数量存在显著的相关性,表明AOB是硝化作用的主要贡献者。5种自然植被条件下AOA的amoA基因拷贝数占泉古菌16S rRNA基因的比例都小于1%(0.01%～0.64%),在农业利用方式下上升到5.32%,表明并非所有泉古菌都具备氨氧化功能基因amoA,氮肥施用可能促进了氨氧化古菌的生长。     

石灰和双氰胺对红壤酸化和硝化作用的影响及其机制

施用石灰是改良酸性土壤的重要措施,但其对土壤硝化作用的增强不仅加速土壤酸化,也增加硝态氮流失风险。传统的硝化抑制剂双氰胺(Dicyandiamide,DCD)能否在石灰改变pH的条件下始终有效抑制硝化是当前红壤区生产中亟需解决的问题。采用短期土壤培养试验,探讨了不同用量石灰与DCD配合施用对土壤酸化和硝化作用的影响及其机制。结果表明:施用一定量的石灰(≤4 g·kg^–1)显著提高土壤pH,通过促进氨氧化细菌的生长以促进硝化作用。在不同pH条件下,DCD对红壤硝化过程均有显著抑制效果。在较高pH(pH 7.0~7.8)条件下,DCD主要通过降低氨氧化细菌的丰度以抑制硝化,而在低pH(pH<6.0)条件下,DCD对氨氧化古菌和氨氧化细菌的丰度均有抑制作用。此外,DCD通过抑制土壤硝化,显著提高了土壤pH。上述结果表明,适宜量(2~4 g·kg^–1)的石灰和DCD结合施用不仅能够减缓红壤酸化,而且能够抑制硝化作用,降低硝态氮的潜在环境风险。     

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.

     

Current insights into the autotrophic thaumarchaeal ammonia oxidation in acidic soils

Recent studies of ammonia-oxidizing archaea (AOA) suggested their significant contributions to global nitrogen cycling, and phylogenetic analysis categorized AOA into a novel archaeal phylum, the Thaumarchaeota. AOA are ubiquitous in terrestrial ecosystems, have unique mechanisms for nitrification, better adaptation to low-pH pressures, and strikingly lower ammonia requirements compared with ammonia-oxidizing bacteria (AOB). Previous perceptions that microbial ammonia oxidation in acidic soils was minimal, and entirely meditated by autotrophic bacteria and occasionally by heterotrophic nitrifiers have been dramatically challenged, and the dominant nitrifying groups urgently called for re-assessment. Controversially, the relative contributions of AOA and AOB to autotrophic ammonia oxidation have been reported to vary in different soils, but ammonia substrate availability, which was largely restricted under acidic conditions, seemed to be the key driver. Theoretically predicted ammonia concentrations in acidic soils below the substrate threshold of AOB and remarkably high ammonia affinity of AOA raised the supposition that thaumarchaea could represent the dominant ammonia-oxidizing group in ammonia-limited acidic environments. Recently, the functional dominance of thaumarchaea over its bacterial counterpart and autotrophic thaumarchaeal ammonia oxidation in acidic soils has been compellingly confirmed by DNA-stable isotope probing (SIP) experiments and the cultivation of an obligate acidophilic thaumarchaeon, Nitrosotalea devanaterra. Here, we review the currently available knowledge concerning the history and progress in our understanding of the ammonia-oxidizing microorganisms (AOB and AOA) and the mechanisms of nitrification in nutrient-depleted acidic soils, present the possible mechanisms shaping the distinct niches of AOA and AOB, and thus strengthen the assumption that AOA dominate over AOB in ammonia oxidation of acidic soils.     

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.     

The effect of soil pH and dicyandiamide (DCD) on N2O emissions and ammonia oxidiser abundance in a stimulated grazed pasture soil

Purpose

Nitrous oxide (N₂O) is a potent greenhouse gas which is mainly produced from agricultural soils through the processes of nitrification and denitrification. Although denitrification is usually the major process responsible for N₂O emissions, N₂O production from nitrification can increase under some soil conditions. Soil pH can affect N₂O emissions by altering N transformations and microbial communities. Bacterial (AOB) and archaeal (AOA) ammonia oxidisers are important for N₂O production as they carry out the rate-limiting step of the nitrification process.

Material and methods

A field study was conducted to investigate the effect of soil pH changes on N₂O emissions, AOB and AOA community abundance, and the efficacy of a nitrification inhibitor, dicyandiamide (DCD), at reducing N₂O emissions from animal urine applied to soil. The effect of three pH treatments, namely alkaline treatment (CaO/NaOH), acid treatment (HCl) and native (water) and four urine and DCD treatments as control (no urine or DCD), urine-only, DCD-only and urine + DCD were assessed in terms of their effect on N₂O emissions and ammonia oxidiser community growth.

Results and discussion

Results showed that total N₂O emissions were increased when the soil was acidified by the acid treatment. This was probably due to incomplete denitrification caused by the inhibition of the assembly of the N₂O reductase enzyme under acidic conditions. AOB population abundance increased when the pH was increased in the alkaline treatment, particularly when animal urine was applied. In contrast, AOA grew in the acid treatment, once the initial inhibitory effect of the urine had subsided. The addition of DCD decreased total N₂O emissions significantly in the acid treatment and decreased peak N₂O emissions in all pH treatments. DCD also inhibited AOB growth in both the alkaline and native pH treatments and inhibited AOA growth in the acid treatment.

Conclusions

These results show that N₂O emissions increase when soil pH decreases. AOB and AOA prefer different soil pH environments to grow: AOB growth is favoured in an alkaline pH and AOA growth favoured in more acidic soils. DCD was effective in inhibiting AOB and AOA when they were actively growing under the different soil pH conditions.     

Impacts of continuous excessive fertilization on soil potential nitrification activity and nitrifying microbial community dynamics in greenhouse system

Purpose

Sampling and analysis of greenhouse soils were conducted in Shouguang, China, to study continuous excessive fertilization effect on nitrifying microbial community dynamics in greenhouse environment.

Materials and methods

Potential nitrification activity (PNA), abundance, and structure of nitrifying microbial communities as well as the correlations with soil properties were investigated.

Results and discussion

Short-term excessive fertilization increased soil nutrient contents and the diversity of nitrifying microbial communities under greenhouse cultivation. However, the abundance and diversity of nitrifying communities decreased greatly due to the increase of soil acidity and salinity after 14 years of high fertilization in greenhouse. There was a significant positive correlation between soil PNA and the abundance of ammonia-oxidizing bacteria (AOB) but not that of ammonia-oxidizing archaea (AOA) in topsoil (0–20 cm) when pH ≥7. Soil PNA and AOB were strongly influenced by soil pH. The groups of Nitrososphaeraceae, Nitrosomonadaceae, and Nitrospiraceae were predominant in the AOA, AOB, and nitrite-oxidizing bacteria (NOB) communities, respectively. Nitrifying community structure was significantly correlated with soil electrical salinity (EC), organic carbon (OC), and nitrate nitrogen (NO₃ ^?–N) content by redundancy analysis (RDA).

Conclusions

Nitrification was predominated by AOB in greenhouse topsoil with high fertilizer loads. Soil salinity, OC, NO₃ ^?–N content, and pH affected by continuous excessive fertilization were the major edaphic factors in shaping nitrifying community structure in greenhouse soils.