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1.
Soil N fertilization stimulates the activity of the soil bacterial species specialized in performing the different steps of the denitrification processes. Different responses of these bacterial denitrifiers to soil N management could alter the efficiency of reduction of the greenhouse gas N2O into N2 gas in cultivated fields. We used next generation sequencing to show how raising the soil N fertility of Canadian canola fields differentially modifies the diversity and composition of nitrite reductase (nirK and nirS) and nitrous oxide reductase (nosZ) gene-carrying denitrifying bacterial communities, based on a randomized complete blocks field experiment. Raising soil N levels increased up to 60% the ratio of the nirK to nirS genes, the two nitrite reductase coding genes, in the Brown soil and up to 300% in the Black soil, but this ratio was unaffected in the Dark Brown soil. Raising soil N levels also increased the diversity of the bacteria carrying the nitrite reductase gene nirK (Simpson index, P = 0.0417 and Shannon index, 0.0181), and changed the proportions of the six dominant phyla hosting nirK, nirS, and nosZ gene-carrying bacteria. The level of soil copper (Cu) and the abundance of nirK gene, which codes for a Cu-dependent nitrite reductase, were positively related in the Brown (P = 0.0060, R2 = 0.48) and Dark Brown (0.0199, R2 = 0.59) soils, but not in the Black soil. The level of total diversity of the denitrifying communities tended to remain constant as N fertilization induced shifts in the composition of these denitrifying communities. Together, our results indicate that higher N fertilizer rate increases the potential risk of nitrous oxide (N2O) emission from canola fields by promoting the proliferation of the mostly adaptive N2O-producing over the less adaptive N2O-reducing bacterial community.  相似文献   

2.
Winter forage grazing systems in New Zealand cause compaction of soil by grazing animals, especially when the soil is wet. However, there is little information on the effects of animal trampling on denitrifiers in soil, despite their importance for N2O production. Here, we report a field study of the abundance of the denitrifying genes nirS, nirK, and nosZ and N2O emissions following the application of dairy cow urine in a free‐draining stony soil. Importantly, we found that simulated animal trampling altered some of the denitrifying microbial communities, thus leading to increased N2O emissions. Over the 111 day measurement period, the abundance of nitrite (NO2?)‐reducing nirS gene copy numbers increased significantly by 87% in the trampled soil with urine (P < 0.01) and increased by 40% in the trampled soil without urine (P < 0.05), but the nirS gene abundance did not change significantly in the nontrampled soil. The abundance of NO2? reducing nirK gene copy numbers was not affected by trampling, but increased significantly following urine application. The abundance of N2O‐reducing nosZ clade I and nosZ clade II gene copy numbers increased significantly in the trampled soil, but did not change significantly in the nontrampled soil. N2O emissions from the trampled soil were about twice that from the nontrampled soil without urine (1.20 and 0.62 kg N2O‐N per ha, respectively) and about eight times greater (6.24 kg N2O‐N per ha) than from nontrampled soil (0.80 kg N2O‐N per ha) when urine was applied. These results strongly suggest that animal trampling during winter forage grazing can have a major impact on denitrifying communities in soil, which in turn stimulate greater denitrification with increased N2O emissions.  相似文献   

3.
Fungi generally dominate microbial biomass in various soils and play critical roles in ecosystem functioning including nutrient cycling, disease ecology and food production. Therefore, fungal denitrification, phenotypically typified by nitrous oxide (N2O) production, presents another avenue other than N mineralization and heterotrophic nitrification for progress to better understand the multiple roles of fungi in sustaining the biosphere. The discovery of N2O production and consequently denitrification in Fusarium oxysporum Schltdl. in early 1970's has led to identification of many taxonomically diverse species of N2O-producing fungi. This review evaluates the current status of knowledge on species composition of fungal denitrifiers and their N2O-producing activity. Here we describe challenges with assessment of fungal N2O-producing activity across genera and suggest prospects for future studies. We also discuss species diversity in order to gain knowledge of important taxonomic and phylogenetic groups mediating N2O production and provide insight on ecological cues associated with fungal N2O production. Currently, the extent to which species phylogeny and the functional trait, i.e. N2O-producing activity, are linked remains to be determined; even so, it is evident that some related taxa exhibit similar N2O production efficacy than distant relatives.  相似文献   

4.
Soil moisture and nitrogen (N) are two important factors influencing N2O emissions and the growth of microorganisms. Here, we carried out a microcosm experiment to evaluate effects of soil moisture level and N fertilizer type on N2O emissions and abundances and composition of associated microbial communities in the two typical arable soils. The abundances and community composition of functional microbes involved in nitrification and denitrification were determined via quantitative PCR (qPCR) and terminal restriction length fragment polymorphism (T-RFLP), respectively. Results showed that N2O production was higher at 90% water-filled pore (WFPS) than at 50% WFPS. The N2O emissions in the two soils amended with ammonium were higher than those amended with nitrate, especially at relatively high moisture level. In both soils, increased soil moisture stimulated the growth of ammonia-oxidizing bacteria (AOB) and nitrite reducer (nirK). Ammonium fertilizer treatment increased the population size of AOB and nirK genes in the alluvial soil, while reduced the abundances of ammonia-oxidizing archaea (AOA) and denitrifiers (nirK and nosZ) in the red soil. Nitrate addition had a negative effect on AOA abundance in the red soil. Total N2O emissions were positively correlated to AOB abundance, but not to other functional genes in the two soils. Changed soil moisture significantly affected AOA rather than AOB community composition in both soils. The way and extent of N fertilizers impacted on nitrifier and denitrifier community composition varied with N form and soil type. These results indicate that N2O emissions and the succession of nitrifying and denitrifying communities are selectively affected by soil moisture and N fertilizer form in the two contrasting types of soil.  相似文献   

5.
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6.
Drained organic forest soils represent a hotspot for nitrous oxide (N2O) emissions, which are directly related to soil fertility, with generally higher emissions from N-rich soils. Highest N2O emissions have been observed in organic forest soils with low pH. The mechanisms for these high emissions are not fully understood. Therefore, the present study was conducted to gain a deeper insight into the underlying mechanisms that drive high N2O emissions from acid soils. Specifically, we investigated the microbial community structure, by phospholipid fatty acid analysis, along a natural pH gradient in an organic forest soil combined with measurements of physico-chemical soil properties. These were then statistically related to site-specific estimates of annual N2O emissions along the same natural pH gradient. Our results indicate that acidic locations with high N2O emissions had a microbial community with an increased fungal dominance. This finding points to the importance of fungi for N2O emissions from acid soils. This may either be directly via fungal N2O production or indirectly via the effect of fungi on the N2O production by other microorganisms (nitrifiers and denitrifiers). The latter may be due to fungal mediated N mineralization, providing substrate for N2O production, or by creating favourable conditions for the bacterial denitrifier community. Therefore, we conclude that enhanced N2O emission from acid forest soil is related, in addition to the known inhibitory effect of low pH on bacterial N2O reduction, to a soil microbial community with increased fungal dominance. Further studies are needed to reveal the exact mechanisms.  相似文献   

7.
This study evaluated the effect of silicate fertilizer on denitrification and associated gene abundance in a paddy soil. A consecutive trial from 2013 to 2015 was conducted including the following treatments: control (CK), mineral fertilizer (NPK), NPK plus sodium metasilicate (NPK + MSF), and NPK plus slag-based silicate fertilizer (NPK + SSF). Real-time quantitative PCR (qPCR) was used to analyze the abundances of nirS, nirK, and nosZ genes. Potential N2O emissions and ammonium and nitrate concentrations were related to the nirS and nirK gene abundance. Compared with the NPK treatments, the addition of a Si fertilizer decreased N2O emission rates and denitrification potential by 32.4–66.6 and 22.0–59.2%, respectively, which were probably related to increased rice productivity, soil Fe availability, and soil N depletion. The abundances of nirS and nirK genes were decreased by 17.7–35.8% and 21.1–43.5% with addition of silicate fertilizers, respectively. Rates of total N2O and N2O from denitrification (DeN2O) emission were positively correlated with the nirS and nirK gene abundance. Nitrate, exchangeable NH4 +, and Fe concentrations were the main factors regulating the nirS and nirK gene abundance. Silicate fertilization during rice growth may serve as an effective approach to decreasing N2O emissions.  相似文献   

8.
The potential denitrification (PD) rate, NO, N2O, and N2 emission were determined after treatment with 50 mg NO3 ??N kg?1 soil using the acetylene inhibition method, and meanwhile abundance of four denitrifying genes (i.e., narG, nirK, norB, nosZ) was also investigated in subtropical soils of China. Soil samples were collected from conifer forest (C), shrub forest, and farmland. These soils were derived from Quaternary red earth and granite. The PD rate and N gas emissions significantly (p?<?0.05) differed between forest and farmland soils; abundance of denitrifying genes was also significantly affected by the land-use change. Correlation and multiple stepwise regression analyses showed that the PD rate was significantly (p?<?0.05) and positively correlated with soil pH but not with soil organic C and total N contents (p?>?0.05). The norB gene copies in farmland soils were significantly higher than in conifer and shrub forest soils (p?<?0.01). Both norB and nosZ gene copies were linearly correlated with soil pH, and the PD rate and N2 emission rate were significantly correlated with the abundance of norB (p?<?0.05). Probably, soil pH affected denitrifiers targeted by the norB gene, thus decreasing the reduction of NO and N2O.  相似文献   

9.
This study assessed the effects that season and tillage practices have on the diversity of nitrous oxide producing bacteria (nitrifiers and denitrifiers) and to relate this to measured N2O fluxes at our field site. Large-scale field plots (1.5 ha) were established in Elora, Ontario in 2000, and managed using conventional tillage (CT) or no-tillage (NT). Each field plot was instrumented with micrometeorological equipment to determine N2O fluxes on a field scale. Soil samples were taken at four time points between the fall of 2005 and the spring of 2006. The diversity of the nitrifier and denitrifier communities was assessed by PCR–denaturing gradient gel electrophoresis (DGGE) using primer pairs targeting the amoA, nirS and nirK gene. Seasonal variation (a combination of soil temperature, available soil moisture, nutrient levels and other potential factors) had the largest influence on the diversity of nitrifier and denitrifier populations; while tillage practice also influenced the diversity of the microbial community at certain time periods. Tillage significantly affected all communities in March and affected denitrifiers on all other dates except for the nirS community in February. Further statistical analysis revealed that diversity of the nitrifying and denitrifying populations was the lowest in February, in frozen soils, and rapidly increased in March, corresponding with spring thaw N2O emissions. Long-term soil nutrient, temperature and N2O data taken at this site added additional information on the dynamics of the nitrogen cycle.  相似文献   

10.
为揭示不同生物硝化抑制剂(BNIs)对红壤性水稻土N2O排放的影响差异及作用机制,通过21 d的土柱淹水培养试验,比较了三种BNIs 1,9-癸二醇(1,9-D)、亚麻酸(LN)和3-(4-羟基苯基)丙酸甲酯(MHPP)与化学合成硝化抑制剂双氰胺(DCD)对土壤N2O排放及相关硝化、反硝化功能基因的影响。结果表明:不同BNIs(1,9-D、LN、MHPP)可以显著平均降低土壤N2O日排放峰值40.1%;1,9-D和MHPP可分别抑制N2O排放总量44.5%和43.9%,而DCD和LN对N2O排放总量没有显著影响。1,9-D和MHPP对AOA(氨氧化古菌)、AOB(氨氧化细菌)硝化菌和nirS、nirK型反硝化菌的调控均有所不同,1,9-D可以同时抑制AOA、AOB和nirS微生物的生长;MHPP仅可以抑制AOA的生长;其中,AOA-amoA和nirS基因丰度与土壤N2O的排放呈显著正相关关系。同时,1,9-D和MHPP均增加了nosZ基因丰度及其与AOA-...  相似文献   

11.
《Applied soil ecology》2010,46(3):201-208
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12.
Pasture soils used for cattle overwintering may represent significant sources of N2O emissions from soils. Therefore, the long-term effect of cattle overwintering on the abundance and activity of a denitrifying community was explored. The study was performed at a cattle overwintering area in South Bohemia (Czech Republic), where three sites differing in the degree of animal impact were selected: severely impacted (SI) and moderately impacted (MI), as well as a control site with no impact (NI). N2O flux measurement and soil sampling were performed in spring and fall of 2005. The activity was measured in terms of potential denitrification activity. Bacterial nirK, nirS and nosZ genes were used as functional markers of the denitrifying communities; abundance was analyzed using a real-time PCR assay. Surprisingly, in situ N2O emissions were the highest in spring at MI and significantly differed from those at SI and NI, while in autumn, rates of emissions generally decreased. In contrast potential denitrification rates were highest at SI, followed by MI, and the lowest at NI. An overall significant shift in N2O/N2 molar ratio was shown in cattle impacted sites. The highest abundance of all genes measured at both sampling times was found at site SI, whereas at site MI increased numbers were observed only in spring. Our results indicate a strong influence of cattle on the abundance as well as the activity of microbes involved in denitrification.  相似文献   

13.
Since the development of effective N2O mitigation options is a key challenge for future agricultural practice, we studied the interactive effect of tillage systems on fertilizer-derived N2O emissions and the abundance of microbial communities involved in N2O production and reduction. Soil samples from 0–10 cm and 10–20 cm depth of reduced tillage and ploughed plots were incubated with dairy slurry (SL) and manure compost (MC) in comparison with calcium ammonium nitrate (CAN) and an unfertilized control (ZERO) for 42 days. N2O and CO2 fluxes, ammonium, nitrate, dissolved organic C, and functional gene abundances (16S rRNA gene, nirK, nirS, nosZ, bacterial and archaeal amoA) were regularly monitored. Averaged across all soil samples, N2O emissions decreased in the order CAN and SL (CAN?=?748.8?±?206.3, SL?=?489.4?±?107.2 μg kg?1) followed by MC (284.2?±?67.3 μg kg?1) and ZERO (29.1?±?5.9 μg kg?1). Highest cumulative N2O emissions were found in 10–20 cm of the reduced tilled soil in CAN and SL. N2O fluxes were assigned to ammonium as source in CAN and SL and correlated positively to bacterial amoA abundances. Additionally, nosZ abundances correlated negatively to N2O fluxes in the organic fertilizer treatments. Soils showed a gradient in soil organic C, 16S rRNA, nirK, and nosZ with greater amounts in the 0–10 than 10–20 cm layer. Abundances of bacterial and archaeal amoA were higher in reduced tilled soil compared to ploughed soils. The study highlights that tillage system induced biophysicochemical stratification impacts net N2O emissions within the soil profile according to N and C species added during fertilization.  相似文献   

14.
Fungal N2O production results from a respiratory denitrification that reduces NO3−/NO2− in response to the oxidation of an electron donor, often organic C. Despite similar heterotrophic nature, fungal denitrifiers may differ from bacterial ones in exploiting diverse resources. We hypothesized that complex C compounds and substances could favor the growth of fungi over bacteria, and thereby leading to fungal dominance for soil N2O emissions. Effects of substrate quality on fungal and bacterial N2O production were, therefore, examined in a 44-d incubation after soils were amended with four different substrates, i.e., glucose, cellulose, winter pea, and switchgrass at 2 mg C g−1 soil. During periodic measurements of soil N2O fluxes at 80% soil water-filled pore space and with the supply of KNO3, substrate treatments were further subjected to four antibiotic treatments, i.e., no antibiotics or soil addition of streptomycin, cycloheximide or both so that fungal and bacterial N2O production could be separated. Up to d 8 when antibiotic inhibition on substrate-induced microbial activity and/or growth was still detectable, bacterial N2O production was generally greater in glucose- than in cellulose-amended soils and also in winter pea- than in switchgrass-amended soils. In contrast, fungal N2O production was more enhanced in soils amended with cellulose than with glucose. Therefore, fungal-to-bacterial contribution ratios were greater in complex than in simple C substrates. These ratios were positively correlated with fungal-to-bacterial activity ratios, i.e., CO2 production ratios, suggesting that substrate-associated fungal or bacterial preferential activity and/or growth might be the cause. Considering substrate depletion over time and thereby becoming limited for microbial N2O production, measurements of soil N2O fluxes were also carried out with additional supply of glucose, irrespective of different substrate treatments. This measurement condition might lead to potentially high rates of fungal and bacterial N2O production. As expected, bacterial N2O production was greater with added glucose than with added cellulose on d 4 and d 8. However, this pattern was broken on d 28, with bacterial N2O production lower with added glucose than with added cellulose. In contrast, plant residue impacts on soil N2O fluxes were consistent over 44-d, with greater bacterial contribution, lower fungal contribution, and thus lower fungal-to-bacterial contribution ratios in winter pea- than in switchgrass-amended soils. Real-time PCR analysis also demonstrated that the ratios of 16S rDNA to ITS and the copy numbers of bacterial denitrifying genes were greater in winter pea- than in switchgrass-amended soils. Despite some inconsistency found on the impacts of cellulose versus glucose on fungal and bacterial leading roles for N2O production, the results generally supported the working hypothesis that complex substrates promoted fungal dominance for soil N2O emissions.  相似文献   

15.
生物质炭在温室气体减排方面具有很大的发展前景,它不仅能实现固碳,对于在大气中停留时间长且增温潜势大的N2O也能发挥积极作用。本研究采用室内厌氧培养试验,按照生物质炭与土壤质量比（0、1%和5%）加入一定量生物质炭,土壤重量含水率控制在20%。利用Robotized Incubation平台实时检测N2O和N2浓度变化,通过测定土壤中反硝化功能基因丰度（nirK、nirS、nosZ）分析生物质炭对N2O消耗的影响及其微生物方面的影响机理。结果表明：经过20 h厌氧培养后,0生物质炭处理的反硝化功能基因丰度（基因拷贝数·g-1）分别为6.80×107（nirK）、5.59×108（nirS）和1.22×108（nosZ）。与0生物质炭处理相比,1%生物质炭处理的nirS基因丰度由最初的2.65×108基因拷贝数·g-1升至7.43×108基因拷贝数·g-1,nosZ基因丰度则提高了一个数量级,由4.82×107基因拷贝数·g-1升至1.50×108基因拷贝数·g-1,然而nirK基因丰度并无明显变化;5%生物质炭处理的反硝化功能基因丰度并未发生显著变化。试验结束时,添加生物质炭处理的N2/（N2O+N2）比值也明显高于0生物质炭处理。相关性分析结果表明,nirS基因丰度和nosZ基因丰度均与N2O浓度在0.01水平上显著相关。试验末期nirS基因丰度和nosZ基因丰度均随着N2O浓度的降低而升高。因此在本试验中,添加1%生物质炭可显著提高nirS和nosZ基因型反硝化细菌的丰度,增大N2/（N2O+N2）比值,促进N2O彻底还原成N2。生物质炭对于N2O主要影响机理是增大了可以还原氧化亚氮的细菌活性,促进完全反硝化。  相似文献   

16.
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17.
Denitrification represents one of the main microbial processes producing the primary and secondary greenhouse gases nitrous oxide (N2O) and nitric oxide (NO) in soils. It is well established that abiotic factors like the soil water content and the availability of nitrogen (N) are key parameters determining the activity of denitrifiers in soils. However, soils differing regarding their characteristics such as the content of Corg, the soil texture or the pH value may respond in specific manners to equivalent changes in soil moisture and N input. Thus, short-term incubation experiments were performed to test and compare the capacity of two contrasting Austrian forest soils to respond to mineral N application at increased soil water contents. Soils from the pristine Rothwald forest (rich in Corg) and the more acidic Schottenwald forest (poor in Corg) were amended with either NH 4 + -N or NO 3 ? -N and were incubated at 40% and 70% water-filled pore space for 4 days. Changes in mineral N pools, nitrite reductase activity and NO and N2O emission rates were measured, and the abundance and structural community composition of the functional group involved in nitrite reduction were analysed via quantitative real-time polymerase chain reaction and terminal restriction fragment length polymorphism analysis of the nirK gene. Rapid and distinct activity responses to increased soil moisture and altered mineral nitrogen availability were observed in two contrasting forest soils. In both soils, nitrogen oxide emission rates were stimulated by N inputs and, depending on the soil moisture status, either NO or N2O emission was prevailing. However, different N cycling processes appeared to predominate in either soil under equivalent treatment. Nitrogen oxide emissions peaked following NO 3 ? application in Schottenwald soils but were the highest after NH 4 + application in Rothwald soils. Denitrifying (nirK) communities differed significantly in Rothwald and Schottenwald soils; however, changes in the community structure were marginal during the short-term incubation. Abundances of nirK genes remained unaffected by N application in either soil. The soil water content affected nirK gene abundances only in Rothwald soil, indicating a distinct reaction of nitrite reducing communities in the two soils.  相似文献   

18.
The process of denitrification has been studied for decades, with current evidence suggesting that an ecosystem's ability to produce and emit N2O is controlled both by transient ‘proximal’ regulators (e.g. temperature, moisture, N availability) as well as distal regulators (e.g. soil type, microbial functional diversity, geography). In this study we use New Zealand soils as a model system to test the impact of distal regulators (i.e. geography) on microbial communities and their N2O emission potential. Using gas chromatography, soil chemical analyses, 16S amplicon sequencing, terminal restriction fragment length polymorphism (T-RFLP) and quantitative PCR (qPCR) on three denitrifier functional genes (nirS, nirK and nosZ), we assessed the factors linked to N2O emissions across a latitudinal gradient. Results show that soil drainage class, soil texture class, and latitude were powerful regulators of both emissions and emission end products (N2 vs. N2O). Mixed models demonstrate that a few variables (including latitude, texture class, drainage class and denitrifier community data [abundance and diversity] amongst others) were enough to predict both the amount and type of gas emitted. In addition we show that microbial community composition (based on 16S rRNA gene sequencing) can also be used to predict both the gas species and quantity emitted.  相似文献   

19.
The production of nitrous oxide (N2O) is a widespread trait in fungi and is of interest because denitrifying fungi lack the N2O reductase gene (nosZ) that regulates N2O reduction to nitrogen gas (N2). The adaptive ability of soil fungi is better than that of bacteria in acidic soils. We investigated the N2O reduction potential, described by the N2O product ratio (RN2O), N2O/(N2O+N2), in soils of different types of fields under crop cultivation with different fertilizer inputs and a bare fallow field with no fertilization as a control. The fungi-to-bacteria abundance ratio (RF/B) was negatively correlated (P < 0.01) with the natural pH of the soil; however, the high value of RF/B measured in vineyards was due to the large inputs of manure. When the denitrification potential was measured at natural pH values of soils, RN2O was negatively correlated (P < 0.01) with soil pH. When the denitrification potential was measured after short-term modifications of soil pH, however, no significant correlation was found between RN2O and the modified pH. Based on stepwise multiple regression analysis, soil pH and residual nitrate (NO3-) were the key factors regulating N2O reduction in soils at natural pH values (R2=0.88, P < 0.001), whereas the key factor was the soil residual NO3- alone (R2=0.83, P < 0.001) when the soil pH was modified. When the effect of the soil chemical properties was weakened, a high RF/B value had the potential (P < 0.01) to affect N2O reduction; however, the role of fungi was offset by the presence of denitrifying bacteria. These results provide evidence that compared to the indirect effects of RF/B, the direct effects of the soil chemical properties have a greater effect on N2O reduction in fertilized soils.  相似文献   

20.
Denitrification is an important part of the nitrogen cycle in the environment, and diverse bacteria, archaea, and fungi are known to have denitrifying ability. Rice paddy field soils have been known to have strong denitrifying activity, but the microbes responsible for denitrification in rice paddy field soils are not well known. Present study analyzed the diversity and quantity of the nitrite reductase genes (nirS and nirK) in a rice paddy field soil, sampled four times in one rice-growing season. Clone library analyses suggested that the denitrifier community composition varied over sampling time. Although many clones were distantly related to the known NirS or NirK, some clones were related to the NirS from Burkholderiales and Rhodocyclales bacteria, and some were related to the NirK from Rhizobiales bacteria. These denitrifiers may play an important role in denitrification in the rice paddy field soil. The quantitative PCR results showed that nirK was more abundant than nirS in all soil samples, but the nirK/nirS ratio decreased after water logging. These results suggest that both diversity and quantity changed over time in the rice paddy field soil, in response to the soil condition.  相似文献   

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