不同施肥方式下土壤氨氧化细菌的群落特征

为了研究长期定位施肥对棕壤中氨氧化细菌（ammonia-oxidizing bacteria,AOB）种群结构多样性和垂直分布特征的影响,本研究采用化学分析、荧光定量PCR（qPCR）和变性梯度凝胶电泳（PCR-DGGE）技术,针对沈阳农业大学试验区不同施肥方式（不施肥、低量无机氮肥、高量无机氮肥、无机氮肥与有机肥配施）下不同土壤深度（0~20 cm、20~40 cm、40~60 cm）的土壤理化性质、AOB丰度及种群多样性进行分析,比较不同施肥方式对土壤AOB种群的影响。结果显示,与不施肥相比,施肥会降低土壤pH,增加土壤铵态氮（70.5%~939.21%）和硝态氮（253.20%~625.48%）含量。随土壤深度增加,土壤pH升高,铵态氮和硝态氮含量除低量无机氮肥处理外,多呈降低趋势。土壤增施氮肥可提高AOB丰度,降低总细菌丰度。其中,0~20 cm土层中AOB丰度较高,且高量无机氮肥处理的AOB数量最高,为9.65×10⁵拷贝数·g^-1（干土）。DGGE图谱分析显示,不同处理下,AOB群落结构多样性指数存在明显差异（P<0.05）,各多样性指数均在表层（0~20 cm）最高,增施氮肥则显著降低AOB的多样性。聚类分析表明,4个施肥处理中,高量无机氮肥处理聚为一类,其他处理则因土壤深度不同而异;3个土壤深度中,除不施肥处理外,所有施肥处理均表现为0~20 cm、20~40 cm土层发生聚类,40~60 cm则明显与其他两层分开。冗余梯度分析（RDA）显示,硝态氮（P=0.027）是造成影响AOB群落结构差异的主要原因。上述研究结果表明,长期定位施肥土壤AOB的数量和群落结构多样性受施肥方式显著影响,并表现出明显的垂直分布特征。与无机氮肥相比,有机无机配施处理有助于改善土壤pH,维持不同土壤深度下AOB群落结构多样性。     

Controls on nitrification in a water-limited ecosystem: experimental inhibition of ammonia-oxidising bacteria in the Patagonian steppe

We studied controls on nitrification in an undisturbed water-limited ecosystem by inhibiting autotrophic nitrifying bacteria in soils with varying levels of vegetative cover. The activity of nitrifying bacteria was disrupted using nitrapyrin, 2-chloro-6-(trichloromethyl)-pyridine, under field conditions in three microenvironments (underneath shrubs, next to grasses and in bare soil). Ammonia-oxidising bacteria were detected by PCR analysis of DNA in soils. The inhibition of nitrification changed the concentrations of NO₃⁻ and NH₄⁺ in the soil, while the microenvironment was most important in determining the response of bacteria to the inhibitor. Nitrapyrin application resulted in a significant (p<0.05) reduction in soil NO₃⁻ concentration (39%) and a significant increase (p<0.001) in soil NH₄⁺ concentration (41%). Untreated bare-soil microenvironments had the lowest concentrations of NH₄⁺ (1.57 μg/g of dry soil) and NO₃⁻ (0.49 μg/g of dry soil) when compared to the other microenvironments, and showed the highest impacts of nitrification inhibition. For example, NH₄⁺ concentrations increased 288% and NO₃⁻ concentrations decreased 60% in inhibited bare-soil microenvironments. In contrast, untreated microenvironments underneath shrubs had the highest levels of NH₄⁺ (10.01 μg/g of dry soil) and NO₃⁻ (0.69 μg/g of dry soil), but showed no significant effects of inhibition of nitrification on soil nitrogen concentrations.     

Relationships between ammonia oxidizers and N2O and CH4 fluxes in agricultural fields with different soil types

Nitrous oxide (N₂O) is a greenhouse gas that contributes to the destruction of stratospheric ozone, and agricultural soil is an important source of N₂O. Aerobic soils are sinks for atmospheric methane (CH₄), a greenhouse gas. Ammonia monooxygenase (AMO) can oxidize CH₄, but CH₄ is mostly oxidized by methane monooxygenase (MMO), and CH₄ oxidation by AMO is generally negligible in the soil. We monitored the N₂O and CH₄ fluxes after urea application in fields containing different soils using an automated sampling system to determine the effects of environmental and microbial factors on the N₂O and CH₄ fluxes. The soil types were Low-humic Andosol (Gleyic Haplic Andosol), yellow soil (Gleyic Haplic Alisol) and gray lowland soil (Entric Fluvisol). Cumulative N₂O emissions from the yellow soil were higher than those from other soil types, although the difference was not significant. The CH₄ uptake level by Andosol was one order of magnitude higher than that by other soils. There were significant relationships between the ammonia oxidation potential, AOB and AOA amoA copy numbers, and the CH₄ uptake. In contrast, the gene copy numbers of methane-oxidizing bacteria (MOB) pmoA were below the detection limit. Our results suggested that the AMOs of AOB and AOA may have more important roles than those previously considered during CH₄ oxidation in agricultural soils treated with N fertilizers.     

脱硫废弃物改良宁夏盐渍化土壤对细菌和氨氧化微生物丰度的影响

为探明脱硫废弃物改良盐渍化土壤对微生物群落的影响效果,在2009~2010年,采用田间试验,施用不同量的脱硫废弃物(0、0.74、1.49、2.25、3.00 kg·m-2),研究了脱硫废弃物对盐渍化土壤细菌、氨氧化细菌和氨氧化古菌的影响。试验结果表明:0~20 cm土层,Ca2+和NO-3-N含量随着施用量增加而增加;土壤p H值、电导率值显著下降。实时荧光定量PCR(q PCR)分析结果表明,微生物丰度随着脱硫废弃物的施用发生变化,但这种变化并不与脱硫废弃物的施用量呈线性关系。在0~20 cm土壤层,施脱硫废弃物使得细菌16S rRNA基因拷贝数处理组显著高于对照组。氨氧化古菌与氨氧化细菌基因拷贝数在T2和T4处理高于其它处理。20~40 cm土层各处理间微生物群落没有显著变化,或没有出现规律的变化趋势。因此,脱硫废弃物增加了土壤细菌和氨氧化功能基因丰度,且对上层土壤影响更为显著。本研究中施用脱硫废弃物1.49 kg·m-2(T2)是引起细菌和氨氧化功能基因丰度增加的施用量。     

Assessing the potential of ammonia oxidizing bacteria to produce nitrous oxide in soils of a high arctic lowland ecosystem on Devon Island, Canada

The contribution of nitrifiers (ammonia-oxidizing bacteria (AOB)) and denitrifiers to nitrous oxide (N₂O) emission from arctic soils remains inconclusive. Based on preliminary experiments, we hypothesized that AOB are the primary producers of N₂O in a high arctic lowland ecosystem on Devon Island, Nunavut, Canada. In part 1 of the study, flux chambers were installed in a catena to determine in situ fluxes of gases (N₂O and carbon dioxide (CO₂)) from 16 June to 13 July 2004. Although fluxes were low, N₂O production occurred in the wettest area of the landscape when ammonium levels were high. As ammonium, but not nitrate, levels declined in the wet sedge meadow, N₂O emissions correspondingly decreased. In part 2, the contribution of nitrification and denitrification to N₂O production was assessed by Acetylene Inhibition Assay and ¹⁵N isotopically enriched incubations. Ammonium fertilization stimulated N₂O emissions to a greater extent than nitrate, and acetylene had a greater impact on N₂O emissions in ammonium-fertilized soils than in nitrate-amended soils. Stable isotope analysis indicated that at 50-55% water filled pore space, nitrification was the dominant (>80%) N₂O emitting process. In part 3, molecular analyses of the two N₂O producing groups indicated the both nitrifiers and denitrifiers did not differ between landforms. Our results suggest nitrifier denitrification is the dominant process occurring in these arctic soils and that the role of denitrifiers in N₂O release from arctic soils needs to be re-evaluated.     

Traceability of ammonia-oxidizing bacteria in compost-treated soils

Composts are increasingly used as environmentally safe biofertilizers in sustainable agriculture all over the world. Although it is well known that composts may contribute to soil vitality and sustainability, and in the enhancement of various soil microbiological processes, little is known about their direct or indirect effects on a microbial-community or population level. Ammonia oxidation by autotrophic ammonia-oxidizing bacteria (AOB) is a key process in agricultural and natural ecosystems and plays an important role in the global nitrogen cycle. Here, we studied the diversity and community composition of ammonia oxidizers in a long-term crop rotation field experiment (>10 years) where four major types of compost (from organic waste, cattle manure, green waste and sewage sludge) had been applied annually. The methods used ranged from PCR-DGGE (denaturing gradient gel electrophoresis) and cloning of 16S rDNA fragments to quantitative real-time PCR. Cluster analysis of DGGE profiles differentiated between the microbial communities of composts, compost-treated soils and mineral-fertilized soils. The community composition of the composts was not reflected in the community composition of the compost-treated soils. Sequencing of screened clones revealed a characteristic AOB community structure for the representative soil sample and the four composts. All AOB-like sequences grouped within the Nitrosospira cluster 3 and 4 and within the Nitrosomonas cluster 6 and 7. The average AOB abundance in compost-treated soils was two times higher than in mineral-fertilized soils (4.3×10⁷ and 1.9×10⁷, respectively). Our data suggest that composts do not leave direct microbial imprints in soils after long-term amendment, but an indirect effect on the AOB community was evident.     

脲酶抑制剂与硝化抑制剂对稻田土壤硝化、反硝化功能菌的影响

[目的]在农业生产中,脲酶抑制剂(urease inhibitor,UI)与硝化抑制剂(nitrification inhibitor,NI)常作为氮肥增效剂来提高肥料利用率。本文研究了在我国南方红壤稻田施用脲酶抑制剂与硝化抑制剂后,土壤中氨氧化细菌(ammonia oxidizing bacteria,AOB)、氨氧化古菌(ammonia-oxidizing archaea,AOA)以及反硝化细菌的丰度以及群落结构的变化特征,旨在揭示抑制剂的作用机理及其对土壤环境的影响。[方法]试验在我国南方红壤稻田进行,共设5个处理:1)不施氮肥(CK);2)尿素(U);3)尿素+脲酶抑制剂(U+UI);4)尿素+硝化抑制剂(U+NI);5)尿素+脲酶抑制剂+硝化抑制剂(U+UI+NI),3次重复。脲酶抑制剂与硝化抑制剂分别为NBPT[N-(n-butyl)thiophosphrictriamide,N-丁基硫代磷酰三胺]和DMPP(3,4-dimethylpyrazole phosphate,3,4-二甲基吡唑磷酸盐)。通过荧光定量PCR(Real-time PCR)研究水稻分蘖期与孕穗期抑制剂对三类微生物标记基因拷贝数的影响,并分析土壤铵态氮、硝态氮与三种菌群丰度的相关性;利用变性梯度凝胶电泳(DenaturingGradient Gel Electrophoresis,DGGE)分析抑制剂对土壤AOB、AOA以及反硝化细菌群落结构的影响,并对优势菌群进行系统发育分析。[结果]1)荧光定量PCR结果表明,施用氮肥对两个时期土壤中AOB的amoA基因与反硝化细菌nirK基因的拷贝数均有显著提高,而对AOA的amoA基因始终没有明显影响;AOB与nirK反硝化细菌的丰度与两个时期的铵态氮含量、分蘖期的硝态氮含量呈极显著正相关,与孕穗期的硝态氮含量相关性不显著;DMPP仅在分蘖期显著减少了AOB的amoA基因拷贝数,表明DMPP主要通过限制AOB的生长来抑制稻田土壤硝化过程;NBPT对三类微生物的丰度无明显影响;2)DGGE图谱表明,在分蘖期与孕穗期,施用氮肥均明显增加了图谱中AOB的条带数,而对AOA却没有明显影响;氮肥明显增加了孕穗期反硝化细菌的条带数;与氮肥的影响相比,抑制剂NBPT与DMPP对AOA、AOB以及反硝化菌的群落结构影响甚微;系统发育分析结果表明,与土壤中AOB的优势菌群序列较为接近的有亚硝化单胞菌和亚硝化螺菌。[结论]在南方红壤稻田中,施入氮肥可显著提高AOB与反硝化细菌的丰度,明显影响两种菌群的群落结构,而AOA较为稳定;NBPT对三类微生物的群落结构丰度无明显影响;硝化抑制剂DMPP可抑制AOB的生长但仅表现在分蘖期,这可能是其缓解硝化反应的主要途径;这也说明二者对土壤生态环境均安全可靠。     

Effect of soil aggregate size and dicyandiamide on N2O emissions and ammonia oxidizer abundance in a grazed pasture soil

Nitrous oxide (N₂O) is a potent greenhouse gas, which is mainly produced from agricultural soils. Ammonia oxidation is the rate‐determining step in N₂O production, and the process is carried out by ammonia oxidizers, bacteria and archaea. Soil aggregate size has been shown to alter soil properties, which affect N₂O emissions and bacterial communities. However, the effect of aggregate size on temporal and total N₂O emissions and ammonia‐oxidizing bacteria (AOB) and archaea (AOA) is not fully understood. This incubation study investigated the effect of three different soil aggregate sizes on N₂O emissions and ammonia oxidizer abundance under high urine‐N concentrations and the effectiveness of a nitrification inhibitor, dicyandiamide (DCD), at reducing N₂O emissions in different aggregate soils. It was found that temporal patterns of N₂O emissions were affected by aggregate size with higher peak emissions in the large and medium aggregates. However, the total emissions were the same due to a ‘switch’ in emissions at day 66, after which smaller aggregates produced higher N₂O emissions. It is suggested that the switch was caused by an increase in aggregate disruption in the small aggregates, following the urine application, due to their higher surface area to volume ratio. AOB and AOA abundances were not significantly affected by aggregate size. DCD was effective in reducing N₂O emissions in all aggregate sizes by an average of 79%. These results suggest that similar ammonia oxidizer abundance is found in soils of different aggregate sizes, and the efficacy of DCD in reducing N₂O emissions was not affected by aggregate size of the soil.     

Characterization of ammonia oxidizing bacteria associated with weeds in a Japanese paddy field using amoA gene fragments

Abstract

Ammonia oxidizing bacteria (AOB) are important microorganisms in rice paddy field ecosystems because they play a key role in the nitrogen (N) cycle by converting ammonia (NH₃) to nitrite (NO^? ₂). In this study, we investigated AOB associated with three types of weeds in a Japanese paddy field (semi-aquatic Echinochloa oryzicola Vasing, floating Lemna paucicostata Hegelm and submerged Najas graminea Delile) using molecular techniques polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and DNA sequencing targeting ammonia monooxygenase (amoA) gene. This work confirmed that rice paddy weeds harbor AOB and that the community composition is different for each type of weed. However, all AOB sequences associated with the tested weeds were closely related to known species of Nitrosospira-like AOB isolated from soil, suggesting that AOB associated with weeds were not specific to weeds and can also be found in the soil. Nitosomanas-like AOB were not detected on any of the weeds tested. In addition, the most dominant AOB strains present in the tested weeds were closely related to Nitrosospira sp. Ka3 and Nitrosospira sp. CT2F. The phylogenetic tree revealed that most of the AOB detected in the present study belonged to amoA cluster 1.     

Humic acids buffer the effects of urea on soil ammonia oxidizers and potential nitrification

Humic acids (HAs) play an important role in the global nitrogen cycle by influencing the distribution, bioavailability, and ultimate fate of organic nitrogen. Ammonium oxidation by autotrophic ammonia-oxidizing bacteria (AOB) is a key process in ecosystems and is limited, in part, by the availability of NH₄⁺. We evaluated the impact of HAs on soil AOB in microcosms by applying urea (1.0%, equal to 10 mg urea/g soil) with 0.1% bHA (biodegraded lignite humic acids, equal to 1 mg/g soil), 0.1% cHA (crude lignite humic acids) or no amendment. AOB population size, ammonium and nitrate concentrations were monitored for 12 weeks after urea and HA application. AOB densities (quantified by real-time PCR targeting the amoA) in the Urea treatments increased about ten-fold (the final abundance: 5.02 × 10⁷ copies (g of dry soil)⁻¹) after one week of incubation and decreased to the initial density after 12 weeks incubation; the population size of total bacteria (quantified by real-time PCR with a universal bacterial probe) decreased from 1.12 × 10¹⁰ to 2.59 × 10⁹ copies (g of dry soil)⁻¹ at week one and fluctuated back to the initial copy number at week 12. In the Urea + bHA and Urea + cHA treatments, the AOB densities were 4 and 6 times higher, respectively, than the initial density of approximately 5.07 × 10⁶ copies (g of dry soil)⁻¹ at week 1 and did not change much up to week 4; the total bacteria density changed little over time. The AOB and total bacteria density of the controls changed little during the 12 weeks of incubation. The microbial community composition of the Urea treatment, based on T-RFLP using CCA (canonical correspondence analysis) and pCCA (partial CCA) analysis, was clearly different from those of other treatments, and suggested that lignite HAs buffered the change in diversity and quantity of total bacteria caused by the application of urea to the soil. We hypothesize that HAs can inhibit the change in microbial community composition and numbers, as well as AOB population size by reducing the hydrolysis rate from urea to ammonium in soils amended with urea.     

Impact of long-term continuous soybean cropping on ammonia oxidizing bacteria communities in the rhizosphere of soybean in Northeast China

Continuous cropping can be a serious problem in Chinese soybean production. This can result in yield reduction, root diseases, and changes in microbial community structure. We studied community structure, clone libraries, and abundance of ammonia oxidizing bacteria (AOB) in soybean fields there were in continuous soybean production for up to 17 years (SC17). Results showed that the potential nitrification rate (PNR) and amoA gene abundance of soybean in continuous cultivation for seven years (SC7) was 0.34 µg NO₃^? g^?1 and 4.71 × 10^?5 amoA gene copies/g dry soil, respectively. These values were lower than other treatments. Phylogenetic affiliation analysis based on blast results of amoA gene clone sequencing showed that the sequences belonged to seven clusters: Cluster 1, Cluster 3b, Cluster 3a.1, Cluster 3a.2, Cluster 9, Cluster 10, and Cluster 4. Correlation of AOB community compositions with environmental factors was performed using canonical correspondence analysis (CCA). Results indicated that the composition of AOB communities in maize–soybean (MS) rotation and continuous cropping of soybean for two years (SC2) were positively related to the PNR of soil, soil moisture, and soil total nitrogen content. Soybean fields continuously cropped for 11 years (SC11) and SC7 fields had AOB community compositions that were negatively related to these factors. The AOB community composition of SC17 was positively correlated to the soil total carbon content of soil. The results in this study indicate that the potential activity and abundance of AOB community in soil significantly changed after seven years continuous cropping compared to other continuous cropping intervals. Cropping systems have important effect on the diversity of functional microorganisms and associated nitrogen cycles.     

The abundance of nitrogen cycle genes amoA and nifH depends on land-uses and soil types in South-Eastern Australia

Nitrogen is a critical nutrient in plant-based primary production systems, therefore measurements of N cycling by microorganisms may add value to agricultural soil monitoring programs. Bacterial-mediated nitrogen cycling was investigated in soils from two broad land-uses (managed and remnant vegetation) across different Soil Orders from three geomorphic zones in Victoria, Australia, by examining the abundance of the genes amoA and nifH using quantitative polymerase chain reaction (qPCR). The aim of the study was to identify parameters influencing bacterial populations possessing the genes nifH and amoA, and examine their distribution at a regional scale across different management treatments. The gene amoA was most abundant in the neutral to slightly alkaline surface soils from Calcarosols in North-West Victoria. There was a highly significant (P < 0.001) interaction between land-use and geomorphic zones in terms of the abundance of amoA. Detection of the gene nifH was site specific with low copy number (less than 100 copies per nanogram of DNA) observed for some strongly acidic surface soil sites in North-East Victoria (Dermosols) and South-West Victoria (Sodosols/Chromosols), while nifH was more abundant in selected Calcarosols of North-West Victoria. The gene amoA was detected across more sites than nifH and was strongly influenced by land-use, with almost consistently greater abundance in managed compared to remnant sites, particularly for North-West and South-West Victoria. The abundance of nifH was not related to land-use, with similar copy numbers observed for both managed and remnant sites at some locations. For the gene nifH, there was no significant interaction between land-use and geomorphic zones, between managed and remnant sites or between the three geomorphic zones. Regression tree analysis revealed a number of likely soil chemical and microbial variables which may act as drivers of gene abundance of amoA and nifH. Variables identified as drivers for amoA included pH, Olsen P, microbial biomass carbon, nitrate and total nitrogen while for nifH the variables were microbial biomass carbon, electrical conductivity, microbial biomass nitrogen, total nitrogen and total potassium. Measures of N cycling genes could be used as an additional indicator of soil health to assess potential ecosystem functions. The spatial scale of the current study demonstrates that a landscape approach may assist soil health monitoring programs by evaluating N cycle gene abundance in the context of the different microbial and chemical conditions related to Soil Order and land-use management.     

Survival of bacterial DNA and culturable bacteria in archived soils from the Rothamsted Broadbalk experiment

Dried soil samples from many sources have been stored in archives world-wide over the years, but there has been little research on their value for studying microbial populations. Samples collected since 1843 from the Broadbalk field experiment on crop nutrition at Rothamsted have been used to document changes in the structure and composition of soils as agricultural practices evolve, also offering an invaluable record of environmental changes from the pre- to post-industrial era in the UK. To date, the microbial communities of these soils have not been studied, in part due to the well-documented drop in bacterial culturability in dried soils. However, modern molecular methods based on PCR amplification of DNA extracted directly from soil do not require bacterial cells to be viable or intact and may allow investigations into the legacy of bacteria that were present at the time of sample collection.

In a preliminary study, to establish if dried soils can provide a historical record of bacterial communities, samples from the Broadbalk soil archive dating back to 1868 were investigated and plots treated with either farmyard manure (FYM) or inorganic fertilizer (NPK) were compared. As anticipated, the processes of air-drying and milling greatly reduced bacterial viability whilst DNA yields declined less and may be preserved by desiccation. A higher proportion of culturable bacteria survived the archiving process in the FYM soil, possibly protected by the increased soil organic matter. The majority of surviving bacteria were firmicutes, whether collected in 2003 or in 1914, but a wide range of genera was detected in DNA extracted from the samples using PCR and DGGE of 16S rRNA genes. Analysis of DGGE band profiles indicated that the two plots maintained divergent populations. Sequence analysis of bands excised from DGGE gels, from a sample collected in 1914, revealed DNA from - and β-proteobacteria as well as firmicutes. PCR using primers specific for ammonia oxidizing bacteria showed similar band profiles across the two treatments in recently collected samples, however older samples from the NPK plot showed greater divergence. Primers specific for the genus Pseudomonas were designed and used in real-time quantitative PCR to indicate that archived soil collected in 1868 contained 10-fold less pseudomonad DNA than fresh soil, representing around 10⁵ genomes g⁻¹ soil. Prior to milling, dramatically less pseudomonad DNA was extracted from recently collected air-dried soil from the NPK compared to the FYM plot; otherwise, the two plots followed similar trends. Overall bacterial abundance, diversity and survival during the archiving process differed in the two soils, possibly due to differences in clay and soil organic matter content. Nevertheless, the results demonstrate that air-dried soils can protect microbial DNA for more than 150 years and offer an invaluable resource for future research.     

The role of ammonium oxidizing communities in mediating effects of an invasive plant on soil nitrification

Invasive plants often benefit from changes that they impose on soil microbes via positive plant–soil feedback, but the mechanisms that underlie these changes, and the legacy of their effects, remain poorly quantified. We investigated the impacts of an invasive annual grass, Microstegium vimineum, on the structure and functioning of soil microbial communities in a multi-year, field-based common garden experiment. Given previous reports that M. vimineum can both elevate nitrification rates in soil and benefit from enhanced nitrate availability, we sought to answer the following questions: 1) Does M. vimineum alter the abundance or composition of soil nitrifying microbial communities (ammonia oxidizing archaea and bacteria, AOA and AOB, respectively)? 2) Are such effects reversible or do soil legacy effects persist after M. vimineum is no longer present? After three years, invaded plots had greater AOA abundances than uninvaded native dominated plots, as well as different AOA community structure. However, after seven years, and following a period of M. vimineum replacement by native plants in the invaded plots, AOA abundances and nitrification rates declined toward levels found in uninvaded plots. Collectively, our results suggest that while the impacts of M. vimineum invasions on nitrogen cycling likely relate to their association with AOA, these effects may not persist if M. vimineum declines over time and native plants and their associated microbes are able to re-establish.     

长期施用含氯化肥对棕壤硝化作用及氨氧化微生物的影响

【目的】氨氧化微生物是氨氧化过程的主要驱动者,氨氧化过程作为硝化作用的限速步骤对氮循环具有重要作用。本研究以沈阳农业大学棕壤含氯化肥长期定位试验的土壤为研究对象,探讨了连续34年施用高氯和低氯化肥对棕壤硝化作用及氨氧化微生物的影响。【方法】该长期试验在等量氮、磷、钾条件下,设置高氯和低氯处理,共8个处理:T1(不施肥);T2(单施尿素);T3(尿素+氯化钾);T4(尿素+过磷酸钙);T5(尿素+过磷酸钙+氯化钾);T6(尿素+磷酸一铵+氯化钾);T7(尿素+氯磷铵+氯化钾);T8(硝酸磷肥+过磷酸钙+氯化钾),T7为高氯处理。采集0—20cm土壤样品,利用荧光定量PCR技术测定氨氧化细菌(AOB)和古菌(AOA)丰度,并结合土壤硝化潜势和基本化学性质,分析长期施用含氯化肥对棕壤硝化作用及氨氧化微生物丰度的影响及影响氨氧化微生物丰度的主要环境因素。【结果】长期施肥降低了土壤pH值,高氯处理降低得最多,显著低于其他处理;高氯处理的土壤硝化潜势也显著低于其他处理,且除高氯处理外,配施磷肥的处理土壤硝化潜势显著高于不施磷处理。各处理土壤中AOA丰度均显著高于AOB,高氯处理土壤中AOA、AOB丰度均显著低于其他处理,土壤硝化潜势与AOA和AOB均呈显著正相关关系。【结论】连续施用高氯化肥34年显著降低了棕壤AOA和AOB丰度,抑制了硝化潜势。该结果可为通过含氯化肥的合理施用来调节土壤AOA和AOB,进而调控土壤氮素循环提供参考。     

土壤亚硝酸气体(HONO)排放过程及其驱动机制

气态亚硝酸(HONO)是大气中氢氧自由基(OH·)的重要来源,直接影响到大气氧化能力和空气质量。通过比较外场测定和模型计算的HONO浓度,发现白天时存在未知的大气HONO来源。研究表明,土壤可以向大气中排放HONO。其机理可能是土壤亚硝态氮和氢离子的化学平衡作用;或土壤夜间吸附和白天解吸附的动态物理化学过程;或氨氧化细菌等微生物的直接排放;也可能是硝化过程中产生的羟胺,在土壤颗粒物等表面的化学反应。因此,土壤HONO排放通量与土壤亚硝态氮浓度、pH、氨氧化细菌丰度、土壤矿物、土壤湿度及C/N值等相关。目前对于土壤HONO排放的研究尚在起步阶段,国内亦少见相关成果报道。本文综述了土壤HONO排放的研究背景、探讨了土壤HONO排放的机理及影响因素,以期为减少氮素损失、提高氮肥利用率、评估氮肥的环境效应及城市空气质量等提供理论依据和科学指导。     

Ammonia-oxidizing communities in agricultural soil incubated with organic waste residues

The impact of organic compounds present in different kinds of organic fertilizers, i.e., anaerobically digested household waste, composted organic household waste, swine manure, and cow manure, on microbial communities in arable soil was investigated using microcosms. Soil was amended with dried residues or organic extracts of the residues and incubated for 12 weeks at 25°C. The microbial community composition was investigated by phospholipid fatty acid (PLFA) analysis, and the community of ammonia-oxidizing bacteria (AOB) was assessed by denaturing gradient gel electrophoresis (DGGE) of 16S rDNA fragments, followed by sequencing. All dried residues increased the AOB activity, determined as potential ammonia oxidation, whereas the organic extracts from the thermophilically digested waste and the swine manure caused a decreased potential activity. However, no differences in the DGGE banding patterns were detected, and the same AOB sequences were present in all samples treated with the residue extracts. Moreover, the PLFA composition showed that none of the residue additions affected the overall microbial community structure in the soil. We conclude that the AOB community composition was not affected by the organic compounds in the fertilizers, although the activity in some cases was.