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.

     

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.

     

Light inhibition of carbon mineralization associated with iron redox processes in calcareous paddy soil

Purpose

The iron redox cycle is closely tied to the fate of carbon in terrestrial ecosystems, especially paddy soils. Varies diurnally and seasonally, light—the crucial environmental factor—may be a fundamental factor elucidating temporal and spatial variabilities of carbon-containing gases emission. The role of sunlight in the iron-mediated carbon cycle, however, has not been fully elucidated. We conduct this study to test the role of light in the iron-mediated carbon cycling.

Materials and methods

In this study, we conducted anaerobic incubation experiments of a calcareous paddy soil in serum vials under alternating dark and light conditions. The dynamic evolution of the carbon and iron contents was evaluated by measuring the CO₂, CH₄, and O₂ concentrations in the headspace of the vials, as well as the water-soluble inorganic carbon, microbial biomass carbon, and HCl-extractable ferrous iron contents in soil slurries. We also analyzed the soil microbial community structure by high-throughput 16S rRNA gene sequencing.

Results and discussion

The results highlighted the positive correlation between carbon mineralization and ferric iron reduction under dark conditions. Under light conditions, however, ferrous iron was oxidized by the O₂ generated via oxygenic photosynthesis of phototrophic bacteria such as Cyanobacteria, along with a decreased production of CO₂, CH₄, and water-soluble inorganic carbon. The abundance of Cyanobacteria positively correlated to O₂ levels and MBC content significantly. Light-induced periodic variations in the redox conditions facilitated carbon fixation in microbial biomass and up to 31.79 μmol g^?1 carbon was sequestrated during 30 days light incubation.

Conclusions

These results indicate that light inhibits the emission of carbon-containing greenhouse gases associated with the iron redox cycle in calcareous paddy soil. Assimilation of inorganic carbon by phototrophs may responsible for the inhibition of carbon mineralization. Our study suggests that procedures allowing more light to reach the soil surface, for instance, reducing the planting density, may mitigate greenhouse gas emissions and promote carbon sequestration in paddy soils.

     

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.     

不同类型水稻土微生物群落结构特征及其影响因素

选取基于我国土壤地理发生分类的不同类型土壤发育的四种水稻土,利用¹⁵N₂气体示踪法测定生物固氮速率,采用实时荧光定量PCR（Real-time PCR）技术测定细菌丰度,通过16S rRNA基因高通量测序分析微生物群落组成和多样性。结果表明：变形菌门（Proteobacteria）、酸杆菌门（Acidobacteria）、绿弯菌门（Chloroflexi）和蓝藻门（Cyanobacteria）是水稻土中优势微生物类群。四种类型土壤发育的水稻土细菌群落结构差异显著（Stress<0.001）,群落结构分异（NMDS1）与土壤pH存在极显著正相关关系（P<0.01）。土壤有机碳和碱解氮含量显著影响水稻土中细菌丰度和群落多样性（P<0.01）。红壤发育的水稻土细菌16S rRNA基因拷贝数显著高于其他三种类型水稻土,但OTU数量、Chao1指数和PD指数均低于其他三种类型水稻土。土壤pH对水稻土生物固氮速率有显著影响（P<0.01）,紫色土发育的水稻土具有最高的生物固氮速率（3.2±0.7 mg×kg^-1×d^-1）,其中优势类群细鞘丝藻属（Leptolyngbya）可能是生物固氮的主要贡献者。研究结果丰富了对水稻土微生物多样性的认识,为通过调控土壤pH和微生物群落组成来提高稻田生物固氮潜力提供了理论依据。     

Long-term submergence of non-methanogenic oxic upland field soils helps to develop the methanogenic archaeal community as revealed by pot and field experiments

The community structure of methanogenic archaea is relatively stable,i.e.,it is sustained at a high abundance with minimal changes in composition,in paddy field soils irrespective of submergence and drainage.In contrast,the abundance in non-methanogenic oxic soils is much lower than that in paddy field soils.This study aimed to describe methanogenic archaeal community development following the long-term submergence of non-methanogenic oxic upland field soils in pot and field experiments.In the pot experiment,a soil sample obtained from an upland field was incubated under submerged conditions for 275 d.Soil samples periodically collected were subjected to culture-dependent most probable number(MPN)enumeration,polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE)analysis of archaeal 16 S r RNA gene,and quantitative PCR analysis of the methyl-coenzyme M reductase alpha subunit gene(mcr A)of methanogenic archaea.The abundance of methanogenic archaea increased from 102 to 103 cells g-1 dry soil and 104 to 107 copies of mcr A gene g-1 dry soil after submergence.Although no methanogenic archaeon was detected prior to incubation by the DGGE analysis,members from Methanocellales,Methanosarcinaceae,and Methanosaetaceae proliferated in the soils,and the community structure was relatively stable once established.In the field experiment,the number of viable methanogenic archaea in a rice paddy field converted from meadow(reclaimed paddy field)was monitored by MPN enumeration over five annual cycles of field operations.Viability was also determined simultaneously in a paddy field where the plow layer soil from a farmer’s paddy field was dressed onto the meadow(dressed paddy field)and an upland crop field converted from the meadow(reclaimed upland field).The number of viable methanogenic archaea in the reclaimed paddy field was below the detection limit before the first cultivation of rice and in the reclaimed upland field.Then,the number gradually increased over five years and finally reached 103–104 cells g-1 dry soil,which was comparable to that in the dressed paddy field.These findings showed that the low abundance of autochthonous methanogenic archaea in the non-methanogenic oxic upland field soils steadily proliferated,and the community structure was developed following repeated and long-term submergence.These results suggest that habitats suitable for methanogenic archaea were established in soil following repeated and long-term submergence.     

间隙灌溉和控释肥施用对稻田土壤产甲烷微生物的影响

间隙灌溉和控释肥施用影响稻田CH_4的产生和排放,然而其微生物机理尚不清楚。本研究通过采集稻季田间原位试验新鲜土样,采用核酸定量技术(qPCR)和末端限制性片段长度多态性(T-RFLP)技术,研究间隙灌溉和控释肥施用对稻田土壤产甲烷微生物群落丰度和结构的影响。结果表明,稻季CH_4排放量与古菌、产甲烷菌(mcr A基因)和甲烷氧化菌(pmo A基因)数量均呈极显著正相关关系(P0.01),而与细菌数量无显著相关性。间隙灌溉显著影响产甲烷菌和甲烷氧化菌数量的季节变化,其中烤田抑制产甲烷菌生长,而对甲烷氧化菌数量没有显著影响。与尿素相比,施用控释肥增加了稻田土壤细菌、古菌和产甲烷菌数量,降低了甲烷氧化菌数量。土壤古菌群落的优势T-RFs片段为184bp和391bp,其中184bp片段的相对丰度随着间隙灌溉的进行由45%~55%降低到23%~30%;而391bp片段则相反,其相对丰度由12%~18%增加到23%~26%。典型相关性分析(CCA)表明间隙灌溉显著影响土壤古菌群落结构(P0.001),而控释肥施用对土壤古菌群落结构没有明显影响。     

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.     

Effects of biochar and 3,4-dimethylpyrazole phosphate (DMPP) on soil ammonia-oxidizing bacteria and nosZ-N2O reducers in the mitigation of N2O emissions from paddy soils

Purpose

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.

Methods

An 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.

Results

The 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.

Conclusion

These 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.

     

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.     

Analysis of the microbial communities in soils of different ages following volcanic eruptions

Volcanism is a primary process of land formation.It provides a model for understanding soil-forming processes and the role of pioneer bacteria and/or archaea as early colonizers in those new environments.The objective of this study was to identify the microbial communities involved in soil formation.DNA was extracted from soil samples from the Llaima volcano in Chile at sites destroyed by lava in different centuries(1640,1751,and 1957).Bacterial and archaeal 16 S r RNA genes were analyzed using quantitative polymerase chain reaction(q PCR)and Illumina Mi Seq sequencing.Results showed that microbial diversity increased with soil age,particularly between the 1751 and 1640 soils.For archaeal communities,Thaumarchaeota was detected in similar abundances in all soils,but Euryarchaeota was rare in the older soils.The analysis of bacterial 16 S r RNA genes showed high abundances of Chloroflexi(37%),Planctomycetes(18%),and Verrucomicrobia(10%)in the youngest soil.Proteobacteria and Acidobacteria were highly abundant in the older soils(16%in 1640 and 15%in 1751 for Acidobacteria;38%in 1640 and 27%in 1751 for Proteobacteria).The microbial profiles in the youngest soils were unusual,with a high abundance of bacteria belonging to the order Ktedonobacterales(Chloroflexi)in the 1957 soil(37%)compared with the 1751(18%)and 1640(7%)soils.In this study,we show that there is a gradual establishment of the microbial community in volcanic soils following an eruption and that specific microbial groups can colonize during the early stages of recovery.     

Changes in proteolytic bacteria in paddy soils in response to organic management

Proteolytic bacterial communities, which mineralize organic nitrogen, play a key role in agricultural systems. In this study, alkaline metalloprotease (apr) gene fragments from proteolytic bacteria were investigated in bulk and rhizosphere paddy soil from four fields under organic management (for 2, 3, 5, and 9 years), and from one field under conventional management (for 2 years). We analyzed the abundance and structure of the proteolytic bacterial communities using real-time quantitative PCR and denaturing gradient gel electrophoresis. Our results showed that the abundance of proteolytic bacteria ranged from 1.57?×?10⁸ to 8.02?×?10⁸?copies/g of soil. In addition, the abundance of the proteolytic bacteria in the paddy soils under organic management was significantly higher than those in the paddy soil under conventional management. Moreover, the gene copy numbers in the rhizosphere soils were significantly higher than those in the bulk soils. The abundance of proteolytic bacteria tended to increase with the duration of organic management, with the highest abundance being found in the soil that had been under organic management for 5 years. However, the proteolytic bacteria communities in the paddy soils were not significantly affected by management practices. Phylogeny analysis showed that all gel bands obtained represented genes from Pseudomonas. Additionally, correlation analysis and canonical correspondence analysis showed that C/N, C, and N were important factors that influenced the abundance and community structure of the proteolytic bacteria. These results suggest that proteolytic bacteria are indicators in organic management systems, depolymerize organic N and hence maintain soil sustainability.

Abbreviations: CM: conventional management; OM: organic management; DGGE: denaturing gradient gel electrophoresis; qPCR: real-time quantitative PCR detecting system; COFCC: China organic food certification center; CCA: canonical correspondence analysis     

Effect of 7-year application of a nitrification inhibitor, dicyandiamide (DCD), on soil microbial biomass, protease and deaminase activities, and the abundance of bacteria and archaea in pasture soils

Purpose

The nitrification inhibitor dicyandiamide (DCD) has been shown to be highly effective in reducing nitrate (NO₃ ^?) leaching and nitrous oxide (N₂O) emissions when used to treat grazed pasture soils. However, there have been few studies on the possible effects of long-term DCD use on other soil enzyme activities or the abundance of the general soil microbial communities. The objective of this study was to determine possible effects of long-term DCD use on key soil enzyme activities involved in the nitrogen (N) cycle and the abundance of bacteria and archaea in grazed pasture soils.

Materials and methods

Three field sites used for this study had been treated with DCD for 7 years in field plot experiments. The three pasture soils from three different regions across New Zealand were Pukemutu silt loam in Southland in the southern South Island, Horotiu silt loam in the Waikato in the central North Island and Templeton silt loam in Canterbury in the central South Island. Control and DCD-treated plots were sampled to analyse soil pH, microbial biomass C and N, protease and deaminase activity, and the abundance of bacteria and archaea.

Results and discussion

The three soils varied significantly in the microbial biomass C (858 to 542 μg C g^?1 soil) and biomass N (63 to 28 μg N g^?1), protease (361 to 694 μg tyrosine g^?1 soil h^?1) and deaminase (4.3 to 5.6 μg NH₄ ⁺ g^?1 soil h^?1) activity, and bacteria (bacterial 16S rRNA gene copy number: 1.64?×?10⁹ to 2.77?×?10⁹ g^?1 soil) and archaea (archaeal 16S rRNA gene copy number: 2.67?×?10⁷ to 3.01?×?10⁸ g^?1 soil) abundance. However, 7 years of DCD use did not significantly affect these microbial population abundance and enzymatic activities. Soil pH values were also not significantly affected by the long-term DCD use.

Conclusions

These results support the hypothesis that DCD is a specific enzyme inhibitor for ammonia oxidation and does not affect other non-target microbial and enzyme activities. The DCD nitrification inhibitor technology, therefore, appears to be an effective mitigation technology for nitrate leaching and nitrous oxide emissions in grazed pasture soils with no adverse impacts on the abundance of bacteria and archaea and key enzyme activities.     

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.     

Effects of straw and its biochar applications on the abundance and community structure of CO2-fixing bacteria in a sandy agricultural soil

Purpose

Crop straw and biochar application can potentially increase carbon sequestration and lead to changes in the microbial community in agricultural soils. Sequestration of CO₂ by autotrophic microorganisms is key to biogeochemical carbon cycling in soil ecosystems. The effects of straw and its biochar, derived from slow pyrolysis, on CO₂ fixation bacteria in sandy soils, remain unclear. Therefore, this study compared the response of abundance and community of CO₂ fixation bacteria to the two straw application methods in a sandy agricultural soil. The overall aim of the study was to achieve an efficient use of straw residues for the soil sustainablility.

Materials and methods

We investigated the soil organic carbon content and autotrophic bacteria over four consecutive years (2014–2018) in a field experiment, including the following four treatments: whole maize straw amendment (S), whole maize straw translated biochar amendment (B), half biochar and half straw amendment (BS), and control (CK) without straw or biochar amendment. The autotrophic bacterial abundance and community structure were measured using molecular methods of real-time PCR, terminal restriction fragment length polymorphisms (T-RFLP), and a clone library targeting the large subunit gene (cbbL) of ribulose-1,5-bisphosphate carboxylase/oxygenase.

Results and discussion

The results showed that the content of soil total organic carbon (TOC), dissolved organic carbon (DOC), and microbial biomass carbon (MBC) in B, S, and BS treatments was significantly increased compared with the CK treatment. Soil TOC and available potassium (AK) in the B treatment significantly increased by 15.4% and 23.3%, respectively, but soil bulk density, DOC, and MBC significantly decreased by 8.5%, 10.6%, and 14.5%, respectively, compared with the S treatment. The abundance of the cbbL gene as well as of the bacterial 16S rRNA gene increased significantly in straw or biochar application treatments as compared to the CK treatment. The B treatment, but not the BS treatment, significantly increased the cbbL gene abundance when compared to the S treatment. No significant differences were observed in the bacterial 16S rRNA gene abundance among the three straw or biochar applications. The application of straw biochar could increase the diversity of the autotrophic bacteria, which also altered the overall microbial composition. Physicochemical properties of the soil, such as soil pH, SOC, and bulk density, can help explain the shift in soil microbial composition observed in the study.

Conclusions

Taken together, our results suggest that straw biochar, rather than straw application, leads to an increase in the abundance and diversity of CO₂-fixing bacteria, which would be advantageous for soil autotrophic CO₂ fixation.

     

Spatio-temporal variability of microbial abundance and community structure in the puddled layer of a paddy soil cultivated with wetland rice (Oryza sativa L.)

The puddled layer of paddy soils represents a highly dynamic environment regarding the spatio-temporal variability of biogeochemical conditions. To study these effects on the abundance and community structure of microbial populations, a rhizotron experiment was conducted throughout an entire growing season of wetland rice. Soil samples were taken from selected areas of the puddled layer (bulk soil, oxidized layer, rhizosphere) at main plant developmental stages such as (i) the initial stage, (ii) tillering, (iii) panicle initiation, (iv) flowering, and (v) maturity. Cell numbers of archaea, bacteria, and selected phyla were assessed by catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH). The structure and diversity of microbial communities was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) along with sequencing of selected bands. Following submergence of the paddy soil, shifts of bacterial community structure were observed in the oxidized layer and the rhizosphere. Members of the β-Proteobacteria became predominant in the rhizosphere at tillering stage and were affiliated with aerobic, iron-oxidizing bacteria of the genus Sideroxydans. Seasonal effects were mainly visible in the rhizosphere, as several phylogenetic subgroups including methanotrophic bacteria showed increased cell numbers at flowering stage. Cell numbers of methanogenic archaea were also highest at flowering stage (bulk soil, rhizosphere) and members of the Methanocellales were identified as predominant archaeal populations in areas of oxic and anoxic conditions. In contrast to bacteria, the communities of archaea in the puddled layer of the studied paddy soil were less influenced by spatio-temporal variations of biogeochemical conditions.     

Soil fertility,enzyme activity,and microbial community structure diversity among different soil textures under different land use types in coastal saline soil

Purpose

This investigation assessed the responses of soil fertility, enzyme activity, and microbial community diversity to soil texture and land use type.

Materials and methods

The tested soils included five soil textures (sandy loam, medium loam, heavy loam, light clay, and medium clay soils) with two land use types (uncultivated and paddy soils) in the coastal zone of Zhejiang Province, China.

Results and discussion

Soil texture had a significant effect on soil pH, electrical conductivity (EC), organic carbon (OC), total nitrogen (TN), available nitrogen (AN), phosphorus (AP) and potassium (AK), catalase and protease activities, total phospholipid fatty acids (PLFAs), bacterial and actinomycetes PLFAs, and microbial diversity (MD). The clay content was significantly positively correlated to soil EC, OC, TN, AN, AP, AK, catalase activity, total PLFAs, bacterial and actinomycetes PLFAs, and MD but significantly negatively associated with soil pH and protease activity. Land use type also had significantly influenced soil pH, EC, OC, TN, AN, AP, AK, catalase, protease and urease activities, total PLFAs, bacterial, actinomycetes, and fungal PLFAs, and MD. The paddy soil had higher OC, TN, AN, AP, catalase, protease and urease activities, total PLFAs, bacterial and actinomycetes PLFAs, and MD but lower soil pH, EC, and AK than the uncultivated soil. The interaction with soil texture and land use type had significantly affected soil pH, EC, OC, TN, AN, AP, AK, catalase and protease activities, total PLFAs, bacterial and actinomycetes PLFAs, and MD.

Conclusions

Soil texture and land use type could be considered important factors in improving soil fertility, enzyme activity, and microbial diversity in coastal saline soils.

     

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.

     

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.