Acetylene Inhibition Technique Research Articles

The ratio of denitrification end-products were influenced by soil pH and clay content across different texture classes in Oklahoma soils

Nitrous oxide (N2O) is a potent greenhouse gas that contributes to stratospheric ozone depletion and global climate change. Soil denitrification has two potential end-products, N2O and dinitrogen (N2), and the ratio of these end-products (N2O:(N2O+N2) or the N2O ratio) is controlled by various factors. This study aims to quantify the influence of soil pH on the ratio of denitrification end-products in Oklahoma soils with different soil textures. Six natural grassland soils encompassing three distinct soil textures were incubated in the laboratory under natural and modified pH with an overall tested pH ranging from 2 to 10. Denitrification end-products were measured in the laboratory using the acetylene inhibition technique and further estimated using a process-based biogeochemical model. Both the laboratory and model results showed that soil pH and texture influenced the ratio of the denitrification end-products. Generally, as soil pH increased the N2O ratio decreased, although both lab and model results indicated that this relationship was not linear. Soil texture may have an indirect effect on the N2O ratio, as two soils of the same texture could have different N2O ratios. However, clay percentage of the soil did show a linear positive correlation with the N2O ratio, suggesting components of soil texture may be more influential than others. Overall, soil pH was a controlling factor in the ratio of denitrification end-products and the newly observed nonlinear relationship warrants further study, particularly when considering its effects in different soil textures.

Direct evidence indicates that revegetation improves organic carbon limitation in sediment denitrification in a eutrophic headwater river

Fluvial sediments are one of the hotspots for river nitrogen removal and denitrification is usually considered the main process of nitrogen removal. The availability of nitrogen in headwater rivers is often a limiting factor for sediment denitrification. However, with the increase of N load caused by human activities, we hypothesized that carbon sources might become a key factor controlling the denitrification of sediments in headwater rivers. Here, we measured the denitrification rate using the acetylene inhibition technique and related environmental factors, including both water and sediment features, throughout a year in a subtropical eutrophic river (1st-3rd order streams). The results of our Pearson correlation and multiple regression analyses supported our hypothesis. We found that C sources had a higher explanatory power than N sources, and sediment total carbon (STC) had the greatest influence on the denitrification rate among the different C sources, and that water carbon sources directly affected the sediment denitrification rates according to a structural equation model (SEM). A degraded river reach was selected for revegetation as part of our study, and we found that revegetation increased water DOC from 19.47 mg l−1 to 24.45 mg l−1, reduced water NO3−-N from 0.61 mg l−1 to 0.35 mg l−1, and significantly increased the sediment denitrification rates by an average of 176%. Our study provides direct evidence that revegetation improves organic carbon limitation in sediment denitrification in a eutrophic headwater river.

Effect of tillage state of paddy soils with heavy metal pollution on the nosZ gene of N2O reductase

Paddy soils are an important source of atmospheric nitrous oxide (N2O). However, numerous studies have focused on N2O production during the soil tillage period, neglecting the N2O production during the dry fallow period. In this study, we conducted an incubation experiment using the acetylene inhibition technique to investigate N2O emission and reduction rates of paddy soil profiles (0-1 m) from Guangdong Province and Jinlin Province in China, with different heavy-metal pollution levels. The abundance and community structures of denitrifying bacteria were determined via quantitative-PCR and Illumina MiSeq sequencing of nosZ, nirK, and nirS genes. Our results showed that the potential N2O emission rate, N2O production rate, and denitrification rate have decreased with increasing soil vertical depth and heavy-metal pollution. More importantly, we found that the functional gene type of N2O reductase switched with the tillage state of paddy soils, which clade Ⅱ nosZ genes were the dominant gene during the tillage period, while clade Ⅰ nosZ genes were the dominant gene during the dry fallow period. The heavy-metal pollution has less effect on the niche differentiation of the nosZ gene. The N2O emission rate was significantly regulated by the genus Bradyhizobium, which contains both N2O reductase and nitrite reductase genes. Our findings suggests that the nosZ gene of N2O reductase can significantly impact the N2O emission from paddy soils.

The influence of soil acidification on N2O emissions derived from fungal and bacterial denitrification using dual isotopocule mapping and acetylene inhibition

Denitrification, as both origins and sinks of N2O, occurs extensively, and is of critical importance for regulating N2O emissions in acidified soils. However, whether soil acidification stimulates N2O emissions, and if so for what reason contributes to stimulate the emissions is uncertain and how the N2O fractions from fungal (ffD) and bacterial (fbD) denitrification change with soil pH is unclear. Thus, a pH gradient (6.2, 7.1, 8.7) was set via manipulating cropland soils (initial pH 8.7) in North China to illustrate the effect of soil acidification on fungal and bacterial denitrification after the addition of KNO3 and glucose. For source partitioning, we used and compared SP/δ18O mapping approach (SP/δ18O MAP) and acetylene inhibition technique combined isotope two endmember mixing model (AIT-IEM). The results showed significantly higher N2O emissions in the acidified soils (pH 6.2 and pH 7.1) compared with the initial soil (pH 8.7). The cumulative N2O emissions during the whole incubation period (15 days) ranged from 7.1 mg N kg−1 for pH 8.7–18.9 mg N kg−1 for pH 6.2. With the addition of glucose, relative to treatments without glucose, this emission also increased with the decrement of pH values, and were significantly stimulated. Similarly, the highest N2O emissions and N2O/(N2O + N2) ratios (rN2O) were observed in the pH 6.2 treatment. But the difference was the highest cumulative N2O + N2 emissions, which were recorded in the pH 7.1 treatment based on SP/δ18O MAP. Based on both approaches, ffD values slightly increased with the acidification of soil, and bacterial denitrification was the dominant pathway in all treatments. The SP/δ18O MAP data indicated that both the rN2O and ffD were lower compared to AIT-IEM. It has been known for long that low pH may lead to high rN2O of denitrification and ffD, but our documentation of a pervasive pH-control of rN2O and ffD by utilizing combined SP/δ18O MAP and AIT-IEM is new. The results of the evaluated N2O emissions by acidified soils are finely explained by high rN2O and enhanced ffD. We argue that soil pH management should be high on the agenda for mitigating N2O emissions in the future, particularly for regions where long-term excessive nitrogen fertilizer is likely to acidify the soils.

The potential importance of soil denitrification as a major N loss pathway in intensive greenhouse vegetable production systems

BackgroundAbout 30 % of vegetables in China are produced in intensively managed greenhouses comprising flood irrigation and extreme rates of nitrogen fertilizers. Little is known about denitrification N losses.MethodsSoil denitrification rates were measured by the acetylene inhibition technique applied to anaerobically incubated soil samples. Four different greenhouse management systems were differentiated: Conventional flood irrigation and over-fertilization (CIF, 800 kg N ha−1, 460 mm); CIF plus straw incorporation (CIF+S, 889 kg N ha−1, 460 mm); Drip fertigation with reduced fertilizer application rates (DIF, 314 kg N ha−1, 190 mm); DIF plus straw incorporation (DIF+S, 403 kg N ha−1, 190 mm). Soil denitrification was measured on nine sampling dates during the growing season (Feb 2019-May 2019) for the top-/ subsoil (0 – 20/ 20- 40 cm) and on three sampling dates for deep soils (40-60/ 80-100 cm). Data was used to constrain N-input-output balances of the different vegetable production systems.ResultsRates of denitrification were at least one magnitude higher in topsoil than in sub- and deep soils. Total seasonal denitrification N losses for the 0 – 40 cm soil layer ranged from 76 (DIF) to 422 kg N ha−1 (CIF+S). Straw addition stimulated soil denitrification in top- and subsoil, but not in deep soil layers. Integrating our denitrification data (0-100 cm) with additional data on N leaching, N2O emissions, plant N uptake, and NH3 volatilization showed, that on average 50 % of added N fertilizers are lost due to denitrification.ConclusionsDenitrification is likely the dominant environmental N loss pathway in greenhouse vegetable production systems. Reducing irrigation and fertilizer application rates while incorporating straw in soils allows the reduction of accumulated nitrate.

Soil denitrifying enzyme activity and its influencing factors in a bamboo forest riparian zone in the upper reaches of the Taihu Lake Basin, China

Soil denitrifying enzyme activity (DEA) was measured by acetylene inhibition technique, along with exploration of factors influencing DEA in a bamboo forest riparian zone in the upper reaches of the Taihu Lake Basin during summer. Our aim was to provide important insights into the assessment of ecological functions of bamboo forest riparian zone on reducing nitrogen pollution in rivers. The results showed that the riparian soil DEA ranged from 6.32 to 23.22 μg N·kg-1·h-1, with a mean value of 14.65 μg N·kg-1·h-1. The vertical distribution (0-40 cm soil profile) of DEA was affected by several factors, such as soil organic carbon (SOC), total nitrogen (TN), nitrate nitrogen (NO3--N), soil water content, and activity of carbon and nitrogen hydrolase, which resulted in decreased DEA with increasing soil depth. The horizontal changes in DEA (at the same soil depth but at different distances from river) was mainly governed by the variation in SOC concentration. In this area, the concentration of soil dissolved organic carbon was relatively low, which might inhibit the soil DEA during summer.

Nitrogen removal through sediment denitrification in the Yangtze Estuary and its adjacent East China Sea: A nitrate limited process during summertime

Nitrogen nutrient surplus is the main cause of a series of environmental problems in the Yangtze Estuary and its adjacent East China Sea (ECS). Denitrification plays an important role in controlling nitrate dynamics and fate in estuarine and coastal environments. We investigated the natural and potential rates of denitrification in the sediments of the Yangtze Estuary and ECS via slurry incubation experiments combined with acetylene inhibition techniques to reveal its contributions to total nitrogen reduction in this hypereutrophic continental shelf area. Key environmental factors, such as the sediment grain size, sediment extractable inorganic nitrogen (NH4+, NO3− and NO2−), sediment organic carbon (SOC), total nitrogen (TN), isotopic compositions (δ13C and δ15N), etc., were also investigated to determine the main factors controlling the denitrification processes. The measured rates of denitrification ranged from 0.39 to 28.49 ng N g−1·h−1. The total nitrogen removed by denitrification in the study area was 3.7 × 1010 g during August. In total, at least 3.3% of the external inorganic nitrogen transported annually into the estuary could be removed by the denitrification processes in the study area. The sediment denitrification rates correlated significantly with the extractable ammonium and δ15N values of surface sediments, indicating that coupled nitrification-denitrification processes may play an important role in nitrogen removal. Almost undetectable levels of nitrate in the sediment further revealed that nitrate supply, regardless of diffusion from the overlying water or production by sediment nitrification processes, is the bottleneck for denitrification.

Roles of bulk and rhizosphere denitrifying bacteria in denitrification from paddy soils under straw return condition

Roles of bulk and rhizosphere denitrifying bacteria in paddy soil denitrification under straw return conditions are rarely discriminated, limiting our understanding on nitrogen biogeochemistry in soils amended with straw. The objective of this study was to explore the microbial mechanisms accounting for the altered rhizosphere and bulk soil denitrification with straw amendment. In this study, straw was added into two representative paddy soils (0.5% w/w) from Yixing (YX) and Taizhou (TZ) in Jiangsu city, China, for rice growth over a 37-day period. Denitrification potentials (net N2O emission, total denitrification, and N2O reduction rates) of rhizosphere and bulk soils at the end of rice growth were measured using an acetylene inhibition technique. In addition, denitrifying bacterial community compositions and functional gene abundances were analyzed using Illumina sequencing and quantitative real-time PCR. For control treatment without straw addition, total denitrification and N2O reduction rates were significantly higher in the rhizosphere than that in bulk soils; however, net N2O emission potentials were similar between rhizosphere and bulk for TZ (22.6–53.3 vs. 9.44–46.6 mg N kg−1) and YX (946 ± 126 vs. 699 ± 350 mg N kg−1). Under straw return condition, N2O emission potentials from the bulk of both soils were significantly elevated, corresponding to the increase in the relative abundance of some taxa of denitrifying bacteria (unnamed environmental samples and unclassified proteobacteria) and increase in nirK, nirS, and nosZ genes. This indicates a positive response of bulk soil denitrifying bacteria to straw addition. In contrast, straw addition produced inhibitive effects on the growth of denitrifying bacteria with decreased nirK, nirS, and nosZ gene abundances in the rhizosphere. This led to decreased N2O emission potentials at the end of incubation for rhizosphere soils, which were 98.2% and 25.2% lower for TZ and YX compared to those of bulk soils. These results suggested that the roles of bulk and rhizosphere denitrifying bacteria in paddy soil denitrification shifted with straw addition, which might inspire further studies to target the denitrification hotspots to effectively mitigate greenhouse emissions at the early period of rice growth.

Residual effects of four-year amendments of organic material on N2O production driven by ammonia-oxidizing archaea and bacteria in a tropical vegetable soil

Organic material (OM) applied to cropland not only enhances soil fertility but also profoundly affects soil nitrogen cycling. However, little is known about the relative contributions of soil ammonia-oxidizing archaea (AOA) and bacteria (AOB) to nitrous oxide (N2O) production during ammonia oxidation in response to the additions of diverse types of OMs in the tropical soil for vegetable production. Herein, the soils were sampled from a tropical vegetable field subjected to 4-year consecutive amendments of straw or manure. All the soils were amended with ammonium sulfate ((NH4)2SO4, applied at a dose of 150 mg N kg−1) and incubated aerobically for four weeks under 50% water holding capacity. 1-octyne or acetylene inhibition technique was used to differentiate the relative contributions of AOA and AOB to N2O production. Results showed that AOA dominated N2O production in soil managements of unfertilized control (CK), chemical fertilization (NPK), and NPK with straw (NPKS), whereas AOB contributed more in soil under NPK with manure (NPKM). Straw addition stimulated AOA-dependent N2O production by 94.8% despite the decreased AOA-amoA abundance. Moreover, manure incorporation triggered both AOA- and AOB-dependent N2O production by 147.2% and 233.7%, respectively, accompanied with increased AOA and AOB abundances. Those stimulating effects were stronger for AOB, owing to its sensitivity to the alleviated soil acidification and decreased soil C/N ratio. Our findings highlight the stimulated N2O emissions during ammonia oxidation by historical OM amendments in tropical vegetable soil, with the magnitude of those priming effects dependent on the types of OM, and appropriate measures need to be taken to counter this challenge in tropical agriculture ecosystems.

Advancement of the Acetylene Inhibition Technique Using Time Series Analysis on Air-Dried Floodplain Soils to Quantify Denitrification Potential

Denitrification in floodplain soils is one key process that determines the buffering capacity of riparian zones in terms of diffuse nitrate pollution. One widely used approach to measure the denitrification potential is the acetylene inhibition technique that requires fresh soil samples. We conducted experiments with air-dried soils using a time series analysis to determine the optimal rewetting period. Thus, air-dried soil samples from six different floodplain areas in Germany were rewetted for 1 to 13days to 100% water-filled pore space. We analyzed nitrogen accumulated as N2O in the top of anaerobic flasks with and without acetylene by gas chromatography after four hours of incubation. We observed an overall optimal rewetting of at least seven days for complete denitrification. We also saw the strong influence of pH and field capacity on the denitrification product ratio; in soils with pH < 7, we hardly assumed complete denitrification, whereas the treatments with pH > 7 achieved stable values after seven days of rewetting. This advanced method provides the opportunity to carry out campaigns with large soil sample sizes on the landscape scale, as samples can be stored dry until measurements are taken.

Biological denitrification in a macrophytic lake: implications for macrophytes-dominated lake management in the north of China

Denitrification plays an important role in nitrogen (N) removal in freshwater ecosystems. Aquatic plants might have an impact on the sediment denitrification of water body, especially in macrophytes-dominated lake; however, there were different opinions about it. Our hypothesis was that the sediment denitrification rates differ significantly in different vegetation zones and seasons because of direct and indirect effect of the aquatic plants. Therefore, we studied sediment denitrification in Dongping Lake, a typical macrophytes-dominated lake located in the north of China. The acetylene inhibition technique was used to quantify the sediment denitrification rates (DRs) in the Phragmites communis (P. communis) zone, aquaculture zone, Potamogeton crispus (P. crispus) zone and mixed vegetation zone in July (summer), October (autumn), December (winter) of 2015 and March (spring) of 2016. The results showed that the average DRs were significantly higher in the P. communis zone (69.0 ± 91.6μmolNm-2h-1) than the mixed vegetation zone (8.70 ± 5.44μmolNm-2h-1), and the average DRs represented significant seasonal difference as in the order of winter (74.5 ± 88.3μmolNm-2h-1) > autumn (15.7 ± 18.6μmolNm-2h-1) ≈ summer (10.7 ± 5.90μmolNm-2h-1) > spring (3.85 ± 1.29μmolNm-2h-1). The DRs generally decreased with the increasing of depth; however, significant increase of DRs with depth were found in certain seasons at the vegetated zones except the non-vegetated zone (the aquaculture zone) indicating the possible rhizosphere effect of aquatic plants on denitrification. The higher DRs and cycling rates of nitrate in the P. communis zone might be related to the larger biomass and oxygen transporting capacity of P. communis than those of the other aquatic plants. Winter peaks of DRs might be attributed to the higher NO3- load and the absence of the plant uptake. The high cycling rates of nitrate in Dongping Lake indicated an enhanced internal N cycling by aquatic plants. Sediment denitrification could remove about 537.7tN every year, which was about 26.5% of annual TN loading in Dongping Lake.

Differences in labile soil organic matter explain potential denitrification and denitrifying communities in a long-term fertilization experiment

Content and quality of organic matter (OM) may strongly affect the denitrification potential of soils. In particular, the impact of soil OM fractions of differing bioavailability (soluble, particulate, and mineral-associated OM) on denitrification remains unresolved. We determined the potential N2O and N2 as well as CO2 production for samples of a Haplic Chernozem from six treatment plots (control, mineral N and NP, farmyard manure - FYM, and FYM + mineral N or NP) of the Static Fertilization Experiment Bad Lauchstädt (Germany) as related to OM properties and denitrifier gene abundances. Soil OM was analyzed for bulk chemical composition (13C-CPMAS NMR spectroscopy) as well as water-extractable, particulate, and mineral-associated fractions. Soils receiving FYM had more total OM and larger portions of labile fractions such as particulate and water-extractable OM. Incubations were run under anoxic conditions without nitrate limitation for seven days at 25 °C in the dark to determine the denitrification potential (N2O and N2) using the acetylene inhibition technique. Abundances of nirS, nirK, and nosZ (I + II) genes were analyzed before and after incubation. The denitrification potential, defined as the combined amount of N released as N2O + N2 over the experimental period, was larger for plots receiving FYM (25.9–27.2 mg N kg−1) than pure mineral fertilization (17.1–19.2 mg N kg−1) or no fertilization (12.6 mg N kg−1). The CO2 and N2O production were well related and up to three-fold larger for FYM-receiving soils than under pure mineral fertilization. The N2 production differed significantly only between all manured and non-manured soils. Nitrogenous gas emissions related most closely to water-extractable organic carbon (WEOC), which again related well to free particulate OM. The larger contribution of N2 production in soils without FYM application, and thus, with less readily decomposable OM, coincided with decreasing abundances of nirS genes (NO2− reductase) and increasing abundances of genes indicating complete denitrifying organisms (nosZ I) during anoxic conditions. Limited OM sources, thus, favored a microbial community more efficient in resource use. This study suggests that WEOC, representing readily bioavailable OM, is a straightforward indicator of the denitrification potential of soils.

The co-regulation of nitrate and temperature on denitrification at the sediment-water interface in the algae-dominated ecosystem of Lake Taihu, China

Sediment denitrification is a dominant mechanism for nitrogen removal and can help to minimize lake eutrophication. However, the spatio-temporal variability of denitrification rates and its controlling factors in sediments of large shallow lakes are poorly understood. In this study, we investigated the controlling factors on the temporal and spatial variability of denitrification rates in Meiliang Bay in Taihu Lake, China, to determine the contribution of denitrification on the total lacustrine nitrogen budget. We collected 18 intact monthly sediment cores and an additional 36 seasonal sediment cores from January 2013 to January 2014. Cores were collected from the inner and outer sections of Meiliang Bay for analysis of denitrification rates and sediment properties. We also collected in situ surface water samples for water quality analysis. Denitrification rates at the sediment-water interface (SWI) were measured using acetylene inhibition techniques and intact sediment core incubation. We used a t test to determine the differences in water quality and sediment properties between the two sites and a one-way ANOVA to identify seasonal differences in denitrification rates, water quality, and sediment properties. We also applied Pearson’s correlation, distance-based redundancy analysis (db-RDA) and random forest model to identify the relationships between denitrification rates and environmental factors. Denitrification rates ranged from 0.76 to 40.94 μmol N m−2 h−1 and 0.13 to 52.55 μmol N m−2 h−1, with annual mean values of 19.97 and 17.15 μmol N m−2 h−1 for the Inner and Outer Bay, respectively. Sediment denitrification rates in the Inner and Outer Bay showed similar seasonal variability, with the highest values in spring and summer and the lowest values in autumn. Nitrate addition was shown to significantly increase denitrification rates in summer and autumn (P < 0.05); however, carbon addition showed no significant influence on denitrification rates in the four seasons. Our results imply that denitrification rates were nitrate-limited in summer and autumn. Distance-based redundancy analysis (db-RDA) and random forest model showed that denitrification rates were mainly determined by nitrate, temperature, and chlorophyll a (Chla), and that nitrate in the water column was the most important predictor of denitrification rates. In general, denitrification rates showed significant seasonal variability in Meiliang Bay due to the co-regulation of both water temperature and nitrate concentrations. The dominance of each controlling factor on denitrification rates varied in different seasons. Based on our calculations, nitrogen removal by denitrification accounted for approximately 10.7% of the total nitrogen input to Taihu Lake. Therefore, we suggest the need for effective measures to reduce external nitrogen inputs of to Lake Taihu to prevent on-going eutrophication.

NirS-type N2O-producers and nosZ II-type N2O-reducers determine the N2O emission potential in farmland rhizosphere soils

Denitrification process in agricultural fields is a large source of nitrous oxide (N2O) emitted to the atmosphere. The rhizosphere soils tend to be the hotspots of denitrification in agricultural soils. Recent studies have reported the important role of nosZ II in eliminating N2O. However, little was known about how these more recently discovered N2O-reducing microorganisms together with other N2O-producers affected the N2O emission in agricultural rhizosphere soils. Here, we compared the potential N2O production rate, the denitrification end-product ratio, and the denitrifier communities between rhizosphere and non-rhizosphere soils of two types of crops in winter and summer. The potential activities were measured by acetylene inhibition technique. QPCR analysis was used to quantify the functional genes. High-throughput sequencing and clone library were conducted to analyze the community structure of denitrifiers. The rhizosphere soils had a higher N2O production potential but lower denitrification end-product ratio (N2O/(N2O+N2)) than the non-rhizosphere soils. The potential N2O production rate was correlated to the nirS-bacteria abundance, especially in terms of the genus Azospirillum. The N2O/(N2O+N2) ratio showed a negative correlation with both the diversity and abundance of the nosZ II-type N2O-reducers. Altogether, we propose that nirS-type N2O-producers and nosZ II-type N2O-reducers can affect the N2O emission in agricultural rhizosphere soils, and enhancement of diversity and abundance of nosZ II-type N2O-reducers may help with the N2O mitigation from upland crops.

Effects of long-term increased N deposition on tropical montane forest soil N2 and N2O emissions

Nitrogen (N) deposition is projected to substantially increase in the tropics over the coming decades, which is expected to lead to enhanced N saturation and gaseous N emissions from tropical forests (via NO, N2O, and N2). However, it is unclear how N deposition in tropical forests influences both the magnitude of gaseous loss of nitrogen and its partitioning into the N2 and N2O loss mechanisms. Here, for the first time, we employed the acetylene inhibition technique and the 15N-nitrate labeling method to quantify N2 and N2O emission rates for long-term experimentally N-enriched treatments in primary and secondary tropical montane forest. We found that during laboratory incubation under aerobic conditions long-term increased N addition of up to 100 kg N ha−1 yr−1 at Jianfengling forest, China, did not cause a significant increase in either N2O or N2 emissions, or N2O/N2. However, under anaerobic conditions, N2O emissions decreased and N2 emissions increased with increasing N addition in the secondary forest. These changes may be attributed to substantially greater N2O reduction to N2 during denitrification, further supported by the decreased N2O/N2 ratio with increasing N addition. No such effects were observed in the primary forest. In both forests, N addition decreased the contribution of denitrification while increasing the contribution of co-denitrification and heterotrophic nitrification to N2O production. Denitrification was the predominant pathway to N2 production (98–100%) and its contribution was unaffected by N addition. Despite the changes in the contributions of denitrification to N2O gas emissions, we detected no change in the abundance of genes associated with denitrification. While the mechanisms for these different responses are not yet clear, our results indicate that the effects of N deposition on gaseous N loss were ecosystem-specific in tropical forests and that the microbial processes responsible for the production of N gases are sensitive to N inputs.

Seasonal variation of potential denitrification rate and enhanced denitrification performance via water-lifting aeration technology in a stratified reservoir—A case study of Zhoucun reservoir

Zhoucun reservoir is one of the major water resources in Zao Zhuang city, northern China. The seasonal distribution of surface sediment denitrification rate and enhanced performance of denitrification via water-lifting aeration technology were explored using the acetylene-inhibition technique. Surface sediment denitrification rates ranged from 2.57 ± 1.32 to 923.90 ± 86.81 nmol N2/(g dw·h), with the highest rates in November (ANOVA, p < 0.05) and significantly low rates in June, July, and August (ANOVA, p < 0.05), mainly because of the seasonal differences in nitrate concentration, water temperature, and sediment total nitrogen (STN). Meanwhile, the N2/(N2+N2O) ratio (83.44–91.70% for the highest sediment denitrification period) indicated that N2 accounted for a majority of denitrification. Correlation analysis between various environmental factors and denitrification was conducted, and nitrate concentration, STN, low molecular weight organic carbon, the number of aerobic denitrifying bacteria, and the environmental parameters of oxidation-reduction potential (ORP), pH, electrical conductivity (EC), and chlorophyll a (Chl-a) presented significant relationships during the entire study period. On the basis of the multiple regression model, nitrate and low molecular weight organic carbon concentration were the most influential factors on denitrification variability. Moreover, the denitrification rates of the surface sediment clearly increased, from 5.28 to 13.22 nmol N2/(g dw·h) to 1117.02–3129.47 nmol N2/(g dw·h), which were higher than those in the non-operating year. This suggests that the denitrification in the sediment system could be enhanced in situ by water-lifting and aeration technology in the reservoir ecosystem.

Effects of nitrate and water content on acetylene inhibition technique bias when analysing soil denitrification rates under an aerobic atmosphere

The acetylene inhibition technique (AIT) is the most widely used indirect method of determining denitrification fluxes. Although AIT bias has long been recognised, the contributions of soil denitrification-related parameters to this bias have not yet been well quantified. Using a direct-N2 method as a baseline, we determined the composition of AIT bias and quantified the effects of soil nitrate (NO3−) and water contents on AIT bias under an aerobic atmosphere. The results showed that the AIT severely underestimated the denitrification rate by 5–26 times and failed to capture the dynamic of denitrification with increasing soil water content. The bias increased with increases in the soil NO3− and water contents. Acetylene-catalysed nitric oxide (NO) oxidation accounted on average for 60% (ranging from 31 to 79%) of the bias. The remaining bias was caused by incomplete acetylene inhibition of nitrous oxide (N2O) reduction. Our results indicate that the AIT method is not recommended for denitrification determination under an aerobic atmosphere, especially for soils with high NO3− and water contents.

Biodegradation of biodiesel and toluene under nitrate-reducing conditions and the impact on bacterial community structure

Biodiesel is a renewable fuel that can be mixed with toluene and be accidentally released into anoxic ecosystems and impact soil microbial communities. Therefore, the aim of the present work was to examine, under nitrate-reduction conditions, the biodegradation of toluene in the presence of two different types of biodiesel (sunflower and rapeseed), and their impact on the bacterial community structure. Sediment samples were spiked individually with toluene, biodiesel, and their blends in laboratory-designed microcosms. Sunflower oil biodiesel was produced in the laboratory, while rapeseed oil biodiesel was a commercial product. Degradation of biodiesels and blends was monitored by directly measuring the substrate or indirectly by determining nitrate removal during the course of the experiment. Denitrification rates were estimated with the acetylene inhibition technique. Denaturing gradient gel electrophoresis was used to assess impacts on the bacterial community structure exposed to biodiesels, blends, and toluene. The results of this study showed that toluene and biodiesel were completely degraded within 10 days. Biodiesel significantly affected the bacterial community structure at a similar magnitude, independently of its origin. Additionally, toluene impacted the bacterial community and denitrification process to a lower extent than biodiesel and a clear decrease in the relative bacterial richness and diversity was shown in samples with biodiesel and blends. To the best of our knowledge, this is one of the first reports describing degradation of biodiesel alone and blends under nitrate-reducing conditions, and also the effects of these compounds on the denitrification process. In addition, due to the recently discovered “oxygenic denitrification” process, the acetylene inhibition technique and nitrous oxide quantification may not be the most adequate tool to estimate denitrification rates. Further detailed studies are advised to understand whether the identified bacterial community shift impacts ecosystem functions. Our results help to understand the biodegradation of toluene, biodiesel, and their blends in sediments under nitrate-reducing conditions and might be important in implementing bioremediation strategies in anoxic environments.

Denitrification and the controlling factors in Yunnan Plateau Lakes (China): Exploring the role of enhanced internal nitrogen cycling by algal blooms.

Denitrification plays an important role in nitrogen (N) removal in freshwater ecosystems. This internal process regulates the fluctuations of N concentration, especially for lakes with high nutrients concentrations and long residence time. Lakes in Yunnan plateau (southwestern China) provide typical cases, while studies in this region have been rare. Therefore, we studied denitrification of two lakes (Lake Dianchi in hypereutrophic state and Lake Erhai in mesotrophic) in this region. We used acetylene inhibition technique to quantify potential denitrification rate (PDR) of these lakes in April and August, 2015 and 2016. PDR of the sediments ranged 0-1.21 μmol/(N·m2·hr), and that of overlying water ranged 0-0.24 μmol/(N·L·hr). Then, we used Least Angle Regression to determine the controlling factors for denitrification. Nutrients controlled PDR from two aspects: providing essential nitrogen sources; and affecting the richness and metabolism of denitrifying bacteria. In April, both aspects limited PDR; while only nitrogen sources limited PDR in August, due to depleted nitrate and enhanced denitrifying bacteria activity. Ammonia was most significant to denitrification, indicating that nitrate from nitrification transported to the bottom of well-mixed lake provide major N source by denitrification. The high PDR and low nitrate concentrate in August were evidence of an enhanced internal N cycling by algal blooms.

Application of two contrasting rice-residue-based biochars triggered gaseous loss of nitrogen under denitrification-favoring conditions: A short-term study based on acetylene inhibition technique

Denitrification is the key microbial process that leads to gaseous loss of soil nitrogen in agricultural lands. Most alarmingly, the dominant gas species could be N2O, which is a much stronger greenhouse gas than CO2. In addition to the primary role as tool for soil carbon sequestration, biochar has the potential to suppress N2O emissions. However, the mechanism for the suppression of N2O emissions by biochar remains elusive. To address this, we performed a short-term incubation experiment targeting the impact of two contrasting biochars on gaseous loss of soil N under denitrification-favoring conditions including high load of NO3− (100 µg N/g soil), anoxia and high moisture content (70% WFPS). The acetylene inhibition technique was adopted to differentiate N losses as N2 and N2O. Two biochars produced from rice chaff (600 °C, pH > 10, C:N 70) or rice husk (300 °C, pH < 5, C:N 54) were applied at two rates (5 and 10% w/w). Results exhibited an increase in gaseous loss of N in all biochar treatments compared with an unamended control. The presence of a positive correlation between total denitrification and CO2 emission rates suggested that biochar derived labile carbon played a pivotal role in triggering the loss of N in gaseous forms. Similarly, the abundances of denitrifying genes nosZ and nirS were considerably higher in biochar treatments, indicating denitrifier’s heterotrophic nature. The biochar pyrolysis conditions and application rates played a decisive role in controlling N2O emissions. Rice chaff biochar, primarily characterized by its alkaline pH, significantly suppressed N2O emissions. Taken together, biochar amendment to agricultural soil can trigger N losses via denitrification and only the alkaline biochar suppressed N2O emissions, probably because of the enhanced activity of N2O reductase.