Ammonia-oxidizing archaea are more important than ammonia-oxidizing bacteria in nitrification and NO3 −-N loss in acidic soil of sloped land

As part of a long-term sloped land use experiment established in 1995 at Taoyuan Agro-ecosystem Research Station (111°26′ E, 28°55′ N) in China, soil samples were collected from three land use types, including cropland (CL), natural forest, and tea plantation. Quantitative polymerase chain reaction and terminal restriction fragment length polymorphism were used to determine the abundance and community composition of amoA-containing bacteria (AOB) and archaea (AOA). The results indicate that land use type induced significant changes in soil potential nitrification rate and community composition, diversity, and abundance of AOB and AOA. Both AOB and AOA community compositions were generally similar between upper and lower slope positions (UP and LP), except within CL. The LP soils had significantly (p?<?0.05) higher diversity and abundance of both AOB and AOA than in the UP. Potential nitrification rate was significantly correlated (p?<?0.05) with diversity and abundance of AOA, but not with AOB. Among land use types, the NO₃ ^? and amoA-containing AOA runoff loss was greatest in CL. Nitrate-N runoff loss was significantly correlated (p?<?0.05) with the loss of AOA amoA copies in the runoff water. Furthermore, relationships between NO₃ ^?-N runoff loss and abundance of AOA but not of AOB at both slope positions were significantly correlated (p?<?0.05). These findings suggest that AOA are more important than AOB in nitrification and NO₃ ^?-N runoff loss in acidic soils across sloped land use types.     

Carbon-iron coupling reduced N2O emissions via promoting the conversion to N2 in paddy soils

The influence of redox reactions involving carbon-iron coupling (organic carbon and iron oxides) on nitrous oxide (N₂O) production in paddy soils remains poorly understood. In this study, two microcosm experiments were conducted to investigate the effects of carbon-iron coupling on N₂O emissions, and the underlying mechanisms were verified using quantitative denitrification functional genes (nirS, nirK, nosZI and nosZII) and high-throughput sequencing. The results showed that ferrihydrite (iron) significantly promoted N₂O-N emissions (p < 0.05) after adding ammonium nitrogen, while glucose (carbon) significantly inhibited N₂O-N emissions (p < 0.05). Carbon-iron coupling significantly decreased N₂O-N emissions (p < 0.05) but did not affect soil total nitrogen loss and increased nitrogen (N₂) emissions. After adding high concentrations of acetylene (10% C₂H₂), the N₂O-N emissions from carbon-iron coupling treatment increased significantly from 6.4 to 11.9 mg N kg⁻¹ (p < 0.05), which confirmed that the carbon-iron coupling reduced the N₂O emissions by promoting the conversion of N₂O to N₂. The mechanisms behind carbon-iron coupling promoting complete denitrification and reducing N₂O emissions were attributed to glucose promoting iron reduction and carbon-iron coupling enhancing the abundance of nosZI (42.7%) and nosZII (16.6%).     

Community composition and activities of denitrifying bacteria from adjacent agricultural soil, riparian soil, and creek sediment in Oregon, USA

We examined denitrifying bacteria from wet soils and creek sediment in an agroecosystem in Oregon, USA that received inputs of nitrogen (N) fertilizer. Our objective was to determine the variation in denitrifying community composition and activities across three adjacent habitats: a fertilized agricultural field planted to perennial ryegrass, a naturally vegetated riparian area, and creek sediment. Using C₂H₂ inhibition, denitrifying enzyme and N₂O-reductase activities were determined in short-term incubations of anaerobic slurries. A key gene in the denitrification pathway, N₂O reductase (nosZ), served as a marker for denitrifiers. Mean denitrifying enzyme activity (DEA) was similar among habitats, ranging from 0.5 to 1.8 μg N g⁻¹ dry soil h⁻¹. However, the ratio of N₂O production, without C₂H₂, to DEA was substantially higher in riparian soil (0.64±0.02; mean±standard error, n=12) than in agricultural soil (0.19±0.02) or creek sediment (0.32±0.03). Mean N₂O-reductase activity ranged from 0.5 to 3.2 μg N g⁻¹ dry soil h⁻¹, with greater activity in agricultural soil than in riparian soil. Denitrifying community composition differed significantly among habitats based on nosZ terminal-restriction fragment length polymorphisms. The creek sediment community was unique. Communities in the agricultural and riparian soil were more closely related but distinct. A number of unique nosZ genotypes were detected in creek sediment. Sequences of nosZ obtained from riparian soil were closely related to nosZ from Bradyrhizobium japonicum. Although nosZ distribution and N₂O-reductase activity differed among habitats, relationships between activity and community composition appeared uncoupled across the agroecosystem.     

Biochar mitigates N2O emissions by promoting complete denitrification in acidic and alkaline paddy soils

Biochar is an efficacious amendment for mitigating nitrous oxide (N₂O) emissions in soils. Nevertheless, the underlying mechanisms responsible for reduced N₂O emissions by biochar in paddy soils remain inadequately elucidated. Here, using two typical paddy soils with contrasting pH values (5.40 and 7.56), the N₂ and N₂O fluxes and the associated functional genes were investigated in soil amended with varying amounts of biochar (0%, 0.5%, and 5%, weight/weight) via soil slurry incubation integrated with the N₂/Ar technique and qPCR analysis. The results showed that N₂O fluxes were significantly (p < 0.05) reduced by 0.65–3.64 times following biochar amendment, concomitant with a significant (p < 0.05) increase in N₂ fluxes (5.47–46.14%) in both acidic and alkaline paddy soils. As a result, the N₂O/(N₂O + N₂) ratios were significantly (p < 0.05) reduced by 1.53–4.65 fold in both soil types. In acidic paddy soils, the enhanced denitrification rates and the decreased N₂O/(N₂O + N₂) ratios exhibited a strong correlation with increased pH values. In alkaline paddy soil, these changes were ascribed to the enhanced nosZ Clade I gene abundance and nosZ/(nirS + nirK) ratio. Our findings reveal that biochar primarily mitigates N₂O emissions in paddy soils by promoting its reduction to N₂.     

Nitrate enhances N2O emission more than ammonium in a highly acidic soil

Purpose

Nitrous oxide (N₂O) is produced naturally in soils through microbial processes of nitrification and denitrification. In recent years, the long-term application of nitrogen-heavy fertilizers has led to the acidification of tea orchard soils with high N₂O emission. The present research aimed at finding out which process (nitrification or denitrification) dominates in N₂O production, whether certain fertilizer managements could reduce N₂O emission, and the effects of fertilizer management on the abundance of functional genes.

Materials and methods

Two nitrification inhibitors, 3, 4-dimethylpyrazole phosphate (DMPP) and dicyandiamide (DCD), combined with different N fertilizers (ammonium sulfate and potassium nitrate) were applied to highly acidic tea orchard soil in an aerobic incubation experiment. Both amoA and nosZ gene abundances from different treatments were determined by quantitative PCR. An anaerobic nitrate effect test was carried out using C₂H₂ inhibition method.

Results and discussion

The application of nitrate fertilizers significantly (P?<?0.05) enhanced total N₂O emission. A linear regression analysis between total N₂O emission and average nitrate contents indicated that denitrification is the dominant source of N₂O in this tea orchard soil. In the anaerobic incubation, no significant difference of N₂O emission was observed between KNO₃ and no KNO₃ treatments before 96 h. Quantitative PCR revealed lower copy numbers of nosZ in nitrate-associated fertilizer-treated soils than the soils from other treatments. Compared with the control, ammonium fertilizers with DCD or DMPP significantly (P?<?0.05) inhibited nitrate production as well as N₂O.

Conclusions

These results showed that denitrification is the dominant source of N₂O in this highly acidic soil. Nitrate addition could significantly inhibit the abundance of nitrous oxide reductase, therefore causing high N₂O emission. The application of ammonium fertilizers with DCD or DMPP could significantly reduce N₂O emission, possibly due to the effective inhibition of nitrate production.     

氮肥水平对稻田细菌群落及N2O排放的影响

作为土壤氮素转化的驱动者,微生物群落结构关系着稻田氮素利用及温室气体N_2O排放等问题。本研究分别基于高通量测序和荧光定量PCR技术,分析了不同氮肥水平[CK(不施氮)、N(施N 180 kg·hm-2)、2/3N(施N 120 kg·hm-2)、1/3N(施N 60 kg·hm-2)]下稻田细菌群落及硝化反硝化关键微生物功能基因丰度的变化。结果显示:氮肥水平提高增加了稻田细菌物种丰富度Chao1指数和群落多样性Shannon指数,改变了细菌群落组成,其中与硝化作用相关的硝化螺菌门Nitrospirae和嗜酸的醋杆菌门Acidobacteria的相对丰度随氮肥水平提高而增加,但甲烷氧化菌Methylosinus的相对丰度随氮肥水平提高而降低。氮肥水平对稻田硝化作用关键微生物氨氧化细菌amo A基因丰度的影响较大,0~5 cm和10~20 cm深度土层中的amo A基因丰度均随氮肥用量增加而提高;反硝化作用关键微生物功能基因nir S、qno B和nos Z的丰度在不施肥处理(CK)中显著低于施肥处理(1/3N、2/3N和N)(P0.05),但1/3N、2/3N和N处理的稻田nir S基因丰度没有明显差异;0~5 cm土层中qno B和nos Z基因丰度存在随氮肥水平提高而增加的趋势,10~20 cm土层中nos Z基因丰度在2/3N和N处理下显著高于1/3N处理(P0.05)。N处理的稻田N_2O排放通量显著高于2/3N及1/3N处理(P0.05),后者又显著高于CK处理(P0.05)。相关分析结果表明稻田N_2O排放通量与0~5 cm土层中硝化螺菌门Nitrospirae相对丰度及10~20 cm土层中amo A基因丰度存在显著相关性(P0.05,n=10)。综上所述,氮肥水平提高增加了稻田细菌群落多样性,促进了稻田N_2O排放,且本研究稻田中硝化作用微生物群落及丰度变化与稻田N_2O排放的关系更为密切。     

Simulated animal trampling of a free‐draining stony soil stimulated denitrifier growth and increased nitrous oxide emissions

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.     

Overwintering management on upland pasture causes shifts in an abundance of denitrifying microbial communities, their activity and N2O-reducing ability

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.     

Incongruous variation of denitrifying bacterial communities as soil N level rises in Canadian canola fields

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.     

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.

     

Straw amendment to paddy soil stimulates denitrification but biochar amendment promotes anaerobic ammonia oxidation

Purpose

Organic matter amendment is usually used to improve soil physicochemical properties and to sequester carbon for counteracting climate change. There is no doubt that such amendment will change microbial activity and soil nitrogen transformation processes. However, the effects of straw and biochar amendment on anammox and denitrification activity and on community structure in paddy soil are unclear.

Materials and methods

We conducted a 30-day pot experiment using rice straw and rice straw biochar to deepen our understanding about the activity, microbial abundance, and community structure associated with soil nitrogen cycling during rice growth.

Results and discussion

Regarding activity, anammox contributed 3.1–8.1% of N₂ production and denitrification contributed 91.9–96.9% of N₂ production; straw amendment resulted in the highest denitrification rate (38.9 nmol N g^?1 h^?1), while biochar amendment resulted in the highest anammox rate (1.60 nmol N g^?1 h^?1). Both straw and biochar amendments significantly increased the hzsB and nosZ gene abundance (p < 0.05). Straw amendment showed the highest nosZ gene abundance, while biochar amendment showed the highest hzsB gene abundance. Phylogenetic analysis of the anammox bacteria 16S rRNA genes indicated that Candidatus Brocadia and Kuenenia were the dominant genera detected in all treatments.

Conclusions

Straw and biochar amendments have different influences on anaerobic ammonia oxidation and denitrification within paddy soil. Our results suggested that the changes in denitrification and anammox rates in the biochar and straw treatments were mainly linked to functional gene abundance rather than microbial community structure and that denitrification played the more major role in N₂ production in paddy soil.

     

Responses of soil nitrous oxide production and abundances and composition of associated microbial communities to nitrogen and water amendment

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.     

Nitrous oxide emissions from a rainfed-cultivated black soil in Northeast China: effect of fertilization and maize crop

Nitrous oxide emission (N₂O) from applied fertilizer across the different agricultural landscapes especially those of rainfed area is extremely variable (both spatially and temporally), thus posing the greatest challenge to researchers, modelers, and policy makers to accurately predict N₂O emissions. Nitrous oxide emissions from a rainfed, maize-planted, black soil (Udic Mollisols) were monitored in the Harbin State Key Agroecological Experimental Station (Harbin, Heilongjiang Province, China). The four treatments were: a bare soil amended with no N (C0) or with 225?kg?N ha^?1 (CN), and maize (Zea mays L.)-planted soils fertilized with no N (P0) or with 225?kg?N ha^?1 (PN). Nitrous oxide emissions significantly (P?<?0.05) increased from 141?±?5?g N₂O-N?ha^?1 (C0) to 570?±?33?g N₂O-N?ha^?1 (CN) in unplanted soil, and from 209?±?29?g N₂O-N?ha^?1 (P0) to 884?±?45?g N₂O-N?ha^?1 (PN) in planted soil. Approximately 75?% of N₂O emissions were from fertilizer N applied and the emission factor (EF) of applied fertilizer N as N₂O in unplanted and planted soils was 0.19 and 0.30?%, respectively. The presence of maize crop significantly (P?<?0.05) increased the N₂O emission by 55?% in the N-fertilized soil but not in the N-unfertilized soil. There was a significant (P?<?0.05) interaction effect of fertilization?×?maize on N₂O emissions. Nitrous oxide fluxes were significantly affected by soil moisture and soil temperature (P?<?0.05), with the temperature sensitivity of 1.73–2.24, which together explained 62–76?% of seasonal variation in N₂O fluxes. Our results demonstrated that N₂O emissions from rainfed arable black soils in Northeast China primarily depended on the application of fertilizer N; however, the EF of fertilizer N as N₂O was low, probably due to low precipitation and soil moisture.     

Relationships between denitrification gene expression,dissimilatory nitrate reduction to ammonium and nitrous oxide and dinitrogen production in montane grassland soils

The montane grassland soils of Europe store significant amounts of nitrogen (N), and climate change might drive their volatilization due to the stimulation of gaseous nitrous oxide (N₂O) and dinitrogen (N₂) losses. Hence, a thorough, mechanistic understanding of the processes responsible for N loss and retention such as denitrification and dissimilatory nitrate reduction to ammonium (DNRA) in these soils is urgently needed. Here we aimed to explore the relationships between denitrifier gene abundance and expression with N₂ and N₂O production and the importance of DNRA versus denitrification in nitrate consumption and N₂O production for typical montane grassland soils of Southern Germany. In a laboratory incubation experiment with glucose and nitrate addition, we combined direct measurements of N₂O and N₂ production with a molecular analysis of the denitrifier communities involved in nitrite, nitric oxide (NO) and N₂O reduction and with the quantification of DNRA. The soils originated from a space-for-time climate change experiment, where intact plant-soil mesocosms were exposed for three years either to ambient conditions at a high elevation site (“HE” control treatment) or to predicted climate change conditions (warming, reduced summer precipitation and reduced winter snow cover) by translocation to lower elevation (“LE” climate change treatment).The abundance (DNA) of cnorB genes was significantly reduced in LE soils, whereas the abundance of nosZ genes did not differ between the HE and LE soils. However, the decreased abundance of cnorB genes unexpectedly resulted in slightly increased rather than decreased potential N₂O emissions. This effect could be explained by the increased levels of cnorB mRNA and, therefore, the higher physiological activity of the NO reducers in the LE soils. In contrast with the DNA levels, the dynamics of the cnorB mRNA levels followed N₂O emission patterns, whereas the nosZ expression was strongly correlated with the N₂ emission (R² = 0.83). The potential rates of DNRA were approximately one-third of the rates of denitrification, and DNRA was not a source for N₂O.We conclude that DNRA significantly competes with denitrification in these soils, thus contributing to N conservation. This work demonstrates that the molecular analysis of nosZ gene expression has great potential to contribute to solving the enigmatic problem of understanding N₂ loss from soil.     

N2O emission mitigation and microbial activity after Biochar and Cao application in a flooded nitrate-rich vegetable soil

Adding easily decomposable organic materials into flooded nitrate-rich soils can effectively decrease the soil nitrate concentration and repair nitrate-rich soil. However, nitrate reduction is usually accompanied with an increase in N₂O emission. This study was conducted to reduce N₂O emission in a nitrate-rich vegetable soil flooded for remediation and amended with biochar. Nitrate-rich vegetable soil was placed in five treatment groups: flooding (F); flooding with rice straw (F?+?RS); flooding with rice straw and 1% biochar (F?+?RS?+?1% biochar); flooding with rice straw and 3% biochar (F?+?RS?+?3% biochar); flooding with rice straw and CaO (F?+?RS?+?CaO). Biochar and CaO reduced the N₂O emission levels relative to the F?+?RS group, with the former being more effective than the latter, achieving reduction of 40.70% (3% biochar) and 17.35% (CaO) of cumulative N₂O emission. The 3% biochar was more effective than the 1% biochar. Regression analysis showed a positive correlation between the abundance of NO reductase gene (norB) and soil N₂O emission flux. In general, biochar and CaO could effectively reduce N₂O emissions from a nitrate-rich vegetable soil during flooding remediation, duo to elevating soil pH and altering denitrifying activity. The norB gene was the most important denitrifying gene driving soil N₂O emission in the remediation.     

Influence of Biochar Addition on the Denitrification Process and N<Subscript>2</Subscript>O Emission in Cd-Contaminated Soil

The aim of this study was to investigate the effect of biochar addition on the denitrification process and N₂O emission in Cd-contaminated soil. Four different biochars, i.e., dairy manure and rice straw pyrolyzed at 350 and 550 °C, respectively, were added into a Cd-contaminated soil and incubation experiments were conducted for 8 weeks. Results showed that Cd had an inhibitory effect on denitrifying reductase enzymes and reduced the abundance of functional genes. On the contrary, amendment with the biochars increased denitrifying enzyme activity and gene abundance, and thus, enhanced the denitrification process. Labile carbon (C) in the biochar-amended soil, which was calculated based on the two-pool exponential model, was the key factor to facilitate this process. As a less important factor, elevated soil pH by biochar addition also increased denitrifying activity as well as the nosZ abundance. Decrease of Cd bioavailability by the biochar addition was beneficial to the denitrification process. Addition of the biochars with higher amount of NO₃ ^?-N, especially the rice straw-derived biochars, increased cumulative N₂O emission by more than ten times relative to the Cd-contaminated soil. With the great amount of labile C and NO₃ ^?-N, the treatment of biochars prepared at 350 °C released the larger amount of CO₂ and N₂O than other treatments. The biochar addition could totally release the heavy metal stress and restore the Cd-contaminated soil in terms of bacterial community.     

生物质炭对土壤N2O消耗的影响及其微生物影响机理

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。     

Denitrification and total nitrogen gas production from forest soils of Eastern China

Reactive forms of nitrogen (Nr) are accumulating at local, regional and global levels largely due to human activities, particularly N-fertilizer production and use as well as fossil fuel combustion. This has resulted in a change in the nitrogen (N) cycle and excess Nr in the environment, which has negative environmental effects. Therefore, characterizing denitrification and the edaphic variables controlling denitrification and its products is the first step in predicting the long-term effects of Nr accumulation. In the present study, six forest soil types in different climatic zones were collected from East China and evaluated for denitrification products following a K¹⁵NO₃ amendment and subsequent incubation. The results showed that denitrification, indicated by production of nitric oxide (NO), nitrous oxide (N₂O) and dinitrogen (N₂), was higher in the studied temperate forest soils than in the studied subtropical and tropical forest soils and was negatively correlated with soil redox potential at the beginning of incubation (r = −0.94, P < 0.01), but not with soil pH. The ratios of NO/total N gas and N₂O/total N gas produced during denitrification varied among the soils, and were generally higher in the subtropical and tropical soils. Spearman's correlation analysis showed that the NO ratio was positively correlated with soil oxidation capacity (OXC) (r = 0.94, P < 0.01) and redox potential at the beginning of incubation (r = 0.86, P < 0.05), but negatively correlated with soil pH (r = −0.83, P < 0.05). The N₂O ratio was not significantly correlated with these edaphic variables, but showed a significant correlation to NO ratio (r = 0.83, P < 0.05). These results suggested that the OXC value might be the key factor affecting denitrification rates in soils. One possible explanation for these effects is that large OXC values would result in a higher level soil redox potential, thus suppressing denitrification and enhancing NO and N₂O ratios during denitrification.