Nitrification Inhibitor Dicyandiamide Research Articles

Enhanced efficiency nitrogen fertilizers (EENFs) can reduce nitrous oxide emissions and maintain high grain yields in a rain-fed spring maize cropping system

ContextEnhanced efficiency nitrogen fertilizers (EENFs) have provided opportunities for the simultaneous mitigation of nitrous oxide (N2O) emissions and increases in crop productions. However, the practice of combining EENFs with optimal fertilization under conditions of misaligned water and fertilizer inputs in rain-fed spring maize systems remains inadequately understood. ObjectiveThis study aims to determine the factors controlling N2O emissions under the soil climatic conditions in rain-fed plastic mulching cropping systems treated with different nitrogen (N) fertilization rates and EENFs, and to determine the most effective N fertilization measure to reduce N2O emissions while maintaining crop yields. MethodsA 3-year field experiment was conducted in a spring maize system located in the semiarid rain-fed region of the Loess Plateau, applying two types of EENFs: a nitrification inhibitor dicyandiamide (DCD) and a slow-release fertilizer (SRF). It comprised five fertilization treatments: unfertilized (CK), conventional N rate (CON, 200 kg N ha−1), optimal N rate (OPT, 160 kg N ha−1), OPT with the addition of DCD (OPT+DCD), and OPT using SRF (OPT-SRF). ResultsThe average annual cumulative N2O emissions over the 3-year experimental period ranged from 0.74 to 1.87 kg N2O-N ha–1. Fertilizer N contributed 26−60% to annual N2O emissions. The highest N2O fluxes (50–161 μg N2O-N m–2 h–1) typically occurred within the initial 10 days following fertilization, likely induced by strong nitrification processes, constituting 12–19% of the annual N2O emissions during this brief period. In comparison to the CON treatment, the OPT, OPT+DCD, and OPT-SRF treatments resulted in significant reductions in annual N2O emissions of 24%, 46%, and 34%, respectively, without causing any significant decrease in maize grain yields. The application of DCD led to increased ammonium (NH4+-N) concentrations while reducing nitrate (NO3–-N). Conversely, using SRF resulted in a decrease in both NH4+-N and NO3–-N concentrations. ConclusionsUsing EENFs at the ideal fertilization rate significantly curtailed yearly N2O emissions without adversely affecting maize yields in a rain-fed plastic mulching spring maize system prevalent in the Loess Plateau. Most N2O emissions occurred post-fertilization, with rainfall events further influencing these emissions. ImplicationsOur findings recommend the incorporation of nitrification inhibitors, such as DCD, as the most effective fertilizer measure for reducing N2O emissions in the rain-fed region of the Loess Plateau.

Reduction of Nitrous Oxide Emissions from Urine Patches from Grazed Dairy Pastures in New Zealand: A Preliminary Assessment of ORUN® as an Alternative to the Use of Nitrification Inhibitor Dicyandiamide (DCD)

Agriculture plays a significant role in economic development and livelihood and is a key contributor to food security and nutrition. However, global concerns regarding the sustainability of the agricultural sector (mainly environmental damage) is linked to agricultural activities such as greenhouse gas (GHG) emissions, water pollution, and loss of biodiversity. The aim of this study was to assess the effectiveness of ORUN® (a formulated agricultural chemical mixture) to reduce N2O emissions from urine patches and to improve pasture yield, pasture N uptake, and soil mineral N concentrations. The field trials were conducted during the spring of 2015 on dairy urine patches at Massey University, New Zealand. Treatments consisted of control nil urine, control nil urine + ProGibb®, urine only, urine + ProGibb®, urine + ORUN®, and urine + ORUN PLUS® replicated four times in a randomized complete block design. At 31 days after treatment (DAT), analysis of soil samples in 0–5 cm soil profiles showed that urine + ProGibb® significantly (p = 0.0041) increased the soil nitrate concentration (121.40 kgN/ha) compared with 48.15 kgN/ha from urine only. The urine + ProGib® treatment produced significantly lower herbage N recovery (35% of applied N) compared with the urine only. Throughout the trial period, the urine patches treated with ProGibb® and ORUN® produced significantly higher N2O fluxes compared with urine only and urine + ORUN PLUS®, as well as higher surface soil nitrate and mineral N concentrations.

Mechanistic insights into mitigating N2O emissions by the nitrification inhibitor dicyandiamide (DCD) in a tropical sandy soil after six years of manure amendment

Organic amendments (OM) can profoundly affect soil nitrous oxide (N2O) emissions via changing nitrogen (N) cycles. However, mechanistic insights into how nitrification inhibitors modulate the responses of soil N2O emissions to successive application of OM are currently lacking. In this study, we performed a laboratory-scale experiment to examine N2O emissions from tropical vegetable soils subjected to six years of chemical fertilization (CF) or chemical fertilization combined with manure application (CFM), and further evaluated the mitigation effectiveness of the nitrification inhibitor dicyandiamide (DCD) under each management regime. Isotopocule mapping (δ15NSPN2O and δ18ON2O/H2O map) showed bacterial nitrification/fungal denitrification contributed 77.4%–88.5% of total N2O production among treatments during the emission peak. The cumulative N2O emissions from the CFM-treated soils were nearly 8-fold of those from the CF-treated soils. CFM stimulated N2O production from bacterial nitrification and denitrification by increasing the abundance of genes linked to nitrifers (AOB and comammox) and denitrifiers (nirK/S and qnorB), respectively. Moreover, DCD decreased cumulative N2O emissions by averagely 73.3%, with better mitigation performance observed in the CFM treatment due to stronger inhibited nitrification and increased abundance of nosZ, and altered bacterial community composition. The 16S sequencing further revealed that adding DCD to CFM-treated soils resulted in declines in abundance of bacterial phylum Actinobacteria and Chloroflexi that positively affected N2O emissions; while the opposite pattern prevailed for Gemmatimonadetes that negatively affected N2O emissions. This study highlights manure application coupled with nitrification inhibitors can achieve the dual goals of enhancing soil fertility while reducing environment risk in tropical agricultural soils.

Non-targeted effects of nitrification inhibitors on soil free-living nitrogen fixation modified with weed management

Biological nitrogen fixation and nitrification inhibitor applications contribute to improving soil nitrogen (N) availability, however, free-living N fixation affected by nitrification inhibitors has not been effectively evaluated in soils under different weed management methods. In this study, the effects of the nitrification inhibitors dicyandiamide (DCD) and 3, 4-dimethylpyrazole phosphate (DMPP) on the nitrogenase, nifH gene，and diazotrophic communities in soils under different weed management methods (AMB, weeds growth without mowing or glyphosate spraying; GS, glyphosate spraying; MSG, mowing and removing weeds and glyphosate spraying; and WM, mowing aboveground weeds) were investigated. Compared to the control counterparts, the DCD application decreased soil nitrogenase activity and nifH gene abundance by 4.5 % and 37.9 %, respectively, under the GS management method, and the DMPP application reduced soil nitrogenase activity by 20.4 % and reduced the nifH gene abundance by 83.4 % under the MSG management method. The application of nitrification inhibitors significantly elevated soil NH4+-N contents but decreased NO3−-N contents, which had adverse impacts on soil nifH gene abundance and nitrogenase activity. The nifH gene abundances were also negatively impacted by dissolved organic N and Geobacter but were positively affected by available phosphorus and diazotrophic community structures. Nitrification inhibitors significantly inhibited Methylocella but stimulated Rhizobiales and affected soil diazotrophic communities. The nitrification inhibitors DCD and DMPP significantly altered soil diazotrophic community structures, but weed management outweighed nitrification inhibitors in reshaping soil diazotrophic community structures. The non-targeted effects of the nitrification inhibitors DMPP and DCD on soil free-living N fixation were substantially influenced by the weed management methods.

Effects of combined natural synergists and chemical inhibitors on yield, nitrogen utilization and balance in wheat/maize rotation system.

Urease inhibitors and nitrification inhibitors can enhance nitrogen (N) fertilizer utilization efficiency and reducing N losses through regulating urea-N transformation. Common urease or nitrification inhibitors, however, are predominantly chemically synthesized and high-cost. Furthermore, their inhibitory effects are mediated by soil pro-perties, climatic conditions, and crop systems. In this study, we conducted a field experiment using natural synergists humic acid/zeolite, along with chemical nitrification inhibitor dicyandiamide (DCD) and their combination to elucidate the impacts of natural synergists combined with chemical inhibitors on annual yield, nitrogen utilization efficiency, soil nitrate-N accumulation, and nitrogen balance within the wheat/maize rotation system. The treatments included no nitrogen fertilizer application (CK), single application of urea (N), urea +DCD (ND), urea + humic acid (NH), urea + zeolite (NP), urea + urease inhibitor N-butylthiophosphoric triamide + DCD (NUD), urea + humic acid + DCD (NHD), and urea + zeolite + DCD (NPD). The results showed that, compared to the treatments NH and NP, the integration of humic acid or zeolite with DCD (NHD and NPD) significantly increased maize yield (11268 and 11397 kg·hm-2) and total annual yield (20494 and 20582 kg·hm-2), which were comparable to those of combined chemical urease and nitrification inhibitors (NUD). The NHD and NPD treatments had higher nitrogen utilization efficiency and lower soil nitrate-N accumulation in 80-100 cm soil layer across all seasons relative to the N treatment, which had no significant difference compared to the NUD treatment. Furthermore, a decline in soil nitrogen surplus by 10.7% and 13.9% was observed when comparing the NHD and NPD treatments with the NH and NP treatments, respectively. These findings suggested that combined humic acid or zeolite and chemical nitrification inhibitors could effectively enhance crop yield and N utilization efficiency and meet the requirements of the green and environmental preservation of modern agriculture.

Insight into functional mechanisms of percarbamide and nitrification inhibitors in degrading fungicide residues and shaping microbial communities in soil-plant systems

Fungicides and nitrogen (N) fertilizers are essential to maintain plant yield in current intensive agriculture. Percarbamide is a novel type of N fertilizer with strong oxidizing property, and the nitrification inhibitor is widely used in agricultural production. It may be feasible to apply percarbamide and nitrification inhibitor as N management to promote fungicide dissipations in soil-plant system. This study quantified the effects of percarbamide and nitrification inhibitor dicyandiamide (DCD) and 3, 4-dimethylpyrazole phosphate (DMPP) on carbendazim residues, and microbial communities of soil-plant systems, and relationships among carbendazim residues, soil and endophytic microbial communities and plant yields were also comprehensively quantified. Compared with the control, the percarbamide significantly reduced soil carbendazim residues by 29.4% but enhanced the lettuce yield by 28.0%. Soil carbendazim residues were significantly and negatively correlated with the soil total N and NO3−-N contents. Soil microbial community structures and co-occurrence networks were more sensitive to N management than their endophytic counterparts. In comparison to the percarbamide alone, the DCD significantly increased the nodes of soil fungal community co-occurrence network which were positively correlated with the plant yield. The DCD outweighed DMPP in increasing the lettuce yield and soil fungal community stability and reshaping soil bacterial community structure. Our study suggested that soil microbial communities were more sensitive to percarbamide and nitrification inhibitor applications than their endophytic counterparts under fungicide pressure and that the DCD outweighed DMPP in reshaping microbial communities. The integrated applications of percarbamide and nitrification inhibitors were promising soil N management strategies to promote fungicide removal and stimulate microbial community in the soil-plant systems.

Measurement of grass uptake of the urease inhibitor NBPT and of the nitrification inhibitor dicyandiamide co-applied with granular urea.

Grass uptake and phytoaccumulation factors of N-(n-butyl) thiophosphoric triamide (NBPT) and dicyandiamide (DCD) were quantified. Following the application of urea fertilizer treated with the inhibitors in Irish grassland, grass samples were collected at 5, 10, 15, 20, and 30day time intervals following five application cycles. Uptake of NBPT by grass was below the limit of quantitation of the analytical method (0.010mg NBPT kg-1). Dicyandiamide concentrations in grass ranged from 0.004 to 28mgkg-1 with the highest concentrations measured on days 5 and 10. A reducing trend in concentration was found after day 15. The DCD phytoaccumulation factor was ranged from 0.004% to 1.1% showing that DCD can be taken up by grass at low levels when co-applied with granular urea. In contrast, NBPT was not detected indicating that grass uptake is unlikely when co-applied with granular urea fertilizer. The contrasting results are likely due to very different longevity of DCD and NBPT along with the much lower rate of NBPT, which is used compared with DCD.

Detection of nitrification inhibitor dicyandiamide: A direct electrochemical approach.

A single run approach for rapid detection of nitrification inhibitor, dicyandiamide (DCD) using electrogenerated chlorine assisted polymerization through azo bond, under acidic conditions and at a preanodized screen printed carbon electrode (SPCE*) is presented. The role of chloride containing support electrolyte in acidic medium along with oxygen functionalities/edge sites are found to be crucial for the successful oxidative polymerization and subsequent adsorption of oxidized products on the electrode surface. The SEM, cyclic voltammetry and X-ray photoelectron spectroscopy studies were used to characterize the polymer film formation. The system exhibited a linear range between 20 and 170μM with a detection limit of 3μM (S/N=3). The method was successfully tested for the detection of DCD in dairy and water samples. Simultaneous detection of DCD in the presence of melamine has also been demonstrated.

Modelling potential human exposure to the nitrification inhibitor dicyandiamide through the environment-food pathway

Dicyandiamide (DCD) is a nitrification inhibitor that effectively reduces nitrogen losses from fertiliser applied to pastures and has the potential to reduce greenhouse gas emissions. Detection of DCD in dairy products in New Zealand led to safety concerns of food products from animals grazing pastures treated with DCD at high application rates (10 kg/ha). This study explores potential risks associated with DCD's application to agricultural grasslands wherein DCD is applied with fertiliser-N at recommended rates. Probabilistic risk assessment method was employed to calculate human Estimated Daily Intake (EDI) of DCD from milk following two application methods (co-application of DCD with granular N-fertiliser and liquid N-fertiliser). The exposure assessment model consists of five separate modules covering application/soil conditions, field conditions, animal feeding, milk production and human exposure/risk to characterise risk in humans. The predicted mean DCD concentration in milk was 0.005 mg DCD/kg milk for the granular method and 0.05 mg DCD/kg milk for the liquid method, respectively. Furthermore, EDI of DCD for both the granular and liquid methods (1 × 10−5 mg and 1 × 10−4 mg DCD/kg body weight/ day, respectively) were well below the European Food Safety Authority set Tolerable Daily Intake limit of 1 mg DCD/kg body weight/ day. Back-calculation revealed that under the conditions of co-application of DCD with granular and liquid nitrogen fertilisers at maximum permissible rates, for EDI to exceed TDI of 1 mg DCD/kg body weight/ day, DCD application rate would need to exceed DCD fertiliser limit (2.25–4.5% by mass of urea nitrogen) set in Commission Regulation (EC) No 1107/2008, and the bio-transfer factor would need to be over 100%. From the sensitivity analysis, for both application types it was found that phytoaccumulation factors, milk intake and the bio-transfer factors are critical factors influencing the EDI. From the present study, the Margin Of Safety values for both application types exceed 1; indicating DCD does not present a significant risk to human health.

Winter wheat responses to enhanced efficiency liquid nitrogen fertilizers in the Canadian Prairies

To evaluate how enhanced efficiency liquid nitrogen (N) fertilizers affect winter wheat ( Triticum aestivum L.) production under irrigated and rain-fed environments, experiments were conducted at two irrigated and five rain-fed sites across the Canadian Prairies from 2013 to 2018 (22 site-years). The N fertilizers included urea ammonium nitrate (UAN) treated with ( i) urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT), ( ii) NBPT plus nitrification inhibitor dicyandiamide, and ( iii) nitrification inhibitor nitrapyrin (Nitrapyrin), as well as untreated UAN and urea, and polymer-coated urea (PCU). All fertilizers were applied by banding 50% at planting and 50% in-crop in early-spring, except PCU, where PCU was applied at planting and urea was applied in early-spring. Nitrous oxide (N2O) emissions and methane (CH4) uptake were measured at one rain-fed site from 2014 to 2017. NBPT increased grain yield by 1.2%–14% and 2.8%–4% under irrigated and rain-fed environments, respectively, relative to all the other N sources except untreated urea in the rain-fed environment. Total N uptake with NBPT was between 0% and 12% higher than the other N sources across irrigated and rain-fed environments. The results suggested that both grain yield and N use efficiency were optimized when UAN contained a urease inhibitor. All liquid enhanced efficiency fertilizers produced grain protein content greater than 11%, except Nitrapyrin under irrigated environments. Data from three site-years indicated that greenhouse gas emissions were unaffected by N source under rain-fed conditions. Liquid UAN with a urease inhibitor may have the most potential to optimize winter wheat production and N use efficiency in the Canadian Prairies.

Winter wheat responses to enhanced efficiency granular nitrogen fertilizer in the Canadian Prairies

Optimizing the timing of nitrogen (N) enhanced efficiency fertilizers (EEFs) may maximize winter wheat ( Triticum aestivum L.) grain yield, protein content, and N-use efficiency (NUE). From 2013 to 2018, experiments were conducted at two irrigated and six rain-fed sites across the Canadian Prairies (24 site-years) to evaluate winter wheat responses to N source and timing/placement effects of EEFs. Nitrogen sources included untreated urea, nitrification inhibitor nitrapyrin treated urea, urease inhibitor N-( n-butyl) thiophosphoric triamide (NBPT) plus nitrification inhibitor dicyandiamide (DCD)-treated urea (NBPT + DCD), and polymer-coated urea (PCU). The N sources were all side-banded at planting, 30% side-banded at planting plus 70% broadcast in-crop late-fall (averaged 38 days after planting; split-applied late-fall), or 30% side-banded at planting plus 70% broadcast in-crop early-spring (averaged 224 days after planting; split-applied early-spring). Nitrous oxide and methane emissions were measured at one rain-fed site to test whether N source and timing/placement influenced CO2-equivalents (CO2-eq; nitrous oxide + methane). Under irrigation, NBPT + DCD consistently produced the highest yields regardless of timing/placement; however, the 80% of the recommended rate caused suboptimal protein responses (≤11%) unless split-application of N was adopted. Untreated urea produced the highest net CO2-eq and yield-scaled CO2-eq emissions, with the highest emissions when urea was split-applied early-spring. To optimize winter wheat production and NUE, we conclude that NBPT + DCD all-banded during seeding operations or split-applied early-spring provided similar and often superior results to other sources, including a more typical system of urea side-banded at the time of seeding.

The win-win effects of nitrification inhibitors on soil-crop systems: Decreasing carbendazim residues but promoting soil bacterial community diversities and stabilities and crop yields

Applying nitrogen (N)-cycling inhibitors is an effective measure to improve N fertilizer utilization efficiency, but the effects of N-cycling inhibitors on fungicide residues in soil-crop systems are unclear. In this study, nitrification inhibitors dicyandiamide (DCD) and 3, 4-dimethylpyrazole phosphate (DMPP) and urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT) were applied into agricultural soils with fungicide carbendazim applications. The soil abiotic properties, carrot yields, carbendazim residues, bacterial communities and their comprehensive relationships were also quantified. Compared to the control treatment, the DCD and DMPP significantly decreased soil carbendazim residues by 96.2% and 96.0%, and the DMPP and NBPT significantly reduced carrot carbendazim residues by 74.3% and 60.3%, respectively. The nitrification inhibitor applications also generated significant and positive effects on carrot yields and soil bacterial community diversities. The DCD application significantly stimulated soil Bacteroidota and endophytic Myxococcota and modified soil and endophytic bacterial communities. Meanwhile, the DCD and DMPP applications also positively stimulated the co-occurrence network edges of soil bacterial communities by 32.6% and 35.2%, respectively. The linear correlation coefficients between soil carbendazim residues and pH, ETSA and NH4+-N contents were − 0.84, − 0.57 and − 0.80, respectively. The nitrification inhibitor applications generated win-win effects on the soil-crop systems by decreasing carbendazim residues but promoting soil bacterial community diversities and stabilities and crop yields.

Combined effects of spent mushroom substrate and dicyandiamide on carbendazim dissipation in soils: Double-edged sword effects and potential risk controls

Repeated and high-dose carbendazim applications have caused serious soil carbendazim contamination, and eco-friendly and economical approaches have been suggested to promote carbendazim removal in agricultural soil. Spent mushroom substrate (SMS) is a special recycled resource after harvesting mushrooms and can be utilized in contaminated soil amendment. The SMS application into agricultural soil might increase antibiotic resistance gene abundances, and the health risks of SMS application might be reduced with reasonable management to adjust the related electron transport of soil nitrification or denitrification. In this study, the SMS and nitrification inhibitor dicyandiamide were used to remediate agricultural soil contaminated with the carbendazim, and the carbendazim contents, soil microbial biomass, activities and community and human disease genes were determined. Compared to the control treatment, the combined applications of SMS and dicyandiamide significantly decreased soil carbendazim content by 38.14% but significantly enhanced soil β-glucosidase, chitinase, arylsulfatase, urease and electron transfer system activities. The relative abundances of Proteobacteria and Actinobacteria were increased by 11.0% and 8.2% with the SMS application, respectively. The carbendazim residues were negatively correlated with the soil pH, electron transfer system activities and relative abundances of Proteobacteria and Actinobacteria. The relative abundances of human disease genes were also dramatically increased with the SMS application, but compared to the SMS alone, extra dicyandiamide application significantly reduced the relative abundances of human disease genes in soils. The SMS applications into fungicide-contaminated soils could generate double-edged sword effects of facilitating fungicide dissipation but leading to potential health risk increase, while applying the dicyandiamide with SMS might be an effective strategy to decrease the negative effect of health risk.

Biochar and Nitrification Inhibitor (Dicyandiamide) Combination Had a Double-Win Effect on Saline-Alkali Soil Improvement and Soybean Production in the Yellow River Delta, China

Salt stress and nutrient deficiency strongly limited the productivity of coastal saline-alkali land in the Yellow River Delta. Biochar has been widely used to improve soil health and promote crop yield, and the positive effects of nitrification inhibitors on fertilizer use efficiency, especially nitrogen, were also verified. However, there were few types of research on the combined application of biochar and nitrification inhibitor dicyandiamide (DCD) on saline-alkali soil of the Yellow River Delta, China. In this study, five treatments, including no nitrogen (CK), normal NPK (N), NPK + 1%biochar (B), NPK + 2%DCD (D), and NPK + 1%biochar + 2%DCD (BD) were set to investigate the single and combined effect of biochar and DCD on nitrogen transform, soil properties, bacterial community structure, and soybean production. Results showed that BD application inhibited nitrification and increased the soil’s nitrate supply at the flowering stage, which reduced nitrogen waste and met the nitrogen demand for soybean growth. Biochar addition increased the soil’s pH and decreased the soil’s electrical conductivities and accelerated the soil’s macroaggregates formation, with the soil’s average mass diameter and geometric average diameter increasing by 78.69% and 30% in B, and 71.29% and 29.34% in BD relative to CK. Positive effects of inhibitors on soybean production were found in increasing soybean yield, hundred-grain weight, aboveground biomass, etc. Proteobacteria was the dominant phylum in the bacterial communities detected, and bacterial community diversity was significantly explained by nitrate content and soil aggregates (p < 0.05). Soil pH and DCD addition mainly influenced the abundance of the bacterial community, especially Actinobacteria. Biochar with DCD could be a feasible fertilization scheme for the coastal saline-alkali land in the Yellow River Delta, China.

Comammox Nitrospira Clade B is the most abundant complete ammonia oxidizer in a dairy pasture soil and inhibited by dicyandiamide and high ammonium concentrations.

The recent discovery of comammox Nitrospira, a complete ammonia oxidizer, capable of completing the nitrification on their own has presented tremendous challenges to our understanding of the nitrification process. There are two divergent clades of comammox Nitrospira, Clade A and B. However, their population abundance, community structure and role in ammonia and nitrite oxidation are poorly understood. We conducted a 94-day microcosm study using a grazed dairy pasture soil amended with urea fertilizers, synthetic cow urine, and the nitrification inhibitor, dicyandiamide (DCD), to investigate the growth and community structure of comammox Nitrospira spp. We discovered that comammox Nitrospira Clade B was two orders of magnitude more abundant than Clade A in this fertile dairy pasture soil and the most abundant subcluster was a distinctive phylogenetic uncultured subcluster Clade B2. We found that comammox Nitrospira Clade B might not play a major role in nitrite oxidation compared to the role of canonical Nitrospira nitrite-oxidizers, however, comammox Nitrospira Clade B is active in nitrification and the growth of comammox Nitrospira Clade B was inhibited by a high ammonium concentration (700 kg synthetic urine-N ha-1) and the nitrification inhibitor DCD. We concluded that comammox Nitrospira Clade B: (1) was the most abundant comammox in the dairy pasture soil; (2) had a low tolerance to ammonium and can be inhibited by DCD; and (3) was not the dominant nitrite-oxidizer in the soil. This is the first study discovering a new subcluster of comammox Nitrospira Clade B2 from an agricultural soil.

Co‐occurrences of soil nitrogen cycling and human‐disease genes following spent mushroom substrate and nitrification inhibitor applications: A strategy for decreasing health risk

AbstractSpent mushroom substrates can be utilized as organic fertilizer in agricultural production but may pose a health risk. Nitrification inhibitors dicyandiamide and 3,4‐dimethylpyrazole phosphate have been used to enhance fertilizer nitrogen utilization efficiency. However, the changes of soil abiotic properties, microbial activities, nitrogen cycling and human‐disease genes, and their comprehensive relationships after the spent mushroom substrate and nitrification inhibitor applications were still unknown. In this study, the spent mushroom substrate and nitrification inhibitor were applied into the agricultural soils at the rates of 2.5% (w/w) and 1.0% of nitrogen content in the spent mushroom substrate, respectively, and then lettuce was planted. Compared with the control, spent mushroom substrate application significantly increased the lettuce biomass, soil enzyme activities and chitinase, protease, nitric‐oxide reductase, nitrogenase, and urease gene abundances. However, the antibiotic resistance and human‐disease gene abundances were also significantly enhanced following the spent mushroom substrate application. Relative to spent mushroom substrate alone, the dicyandiamide application significantly decreased the soil pH and human‐disease gene abundances, although extra dicyandiamide and 3,4‐dimethylpyrazole phosphate enhanced 61.4% and 45.0% of dry biomass of aboveground lettuce. The human‐disease gene abundances were positively correlated with soil pH but were negatively correlated with the Acidobacteria and Streptomyces ratios in soils. Spent mushroom substrate application could generate double‐edged sword effects, and the health risk should be considered in the spent mushroom substrate applications. Extra dicyandiamide application could further promote crop growth while decrease the human‐disease risk from the spent mushroom substrate.

Effects of water regimes on soil N2O, CH4 and CO2 emissions following addition of dicyandiamide and N fertilizer

Water regimes strongly impact soil C and N cycling and the associated greenhouse gases (GHGs, i.e., CO2, CH4 and N2O). Therefore, a study was conducted to examine the impacts of flooding-drying of soil along with application of nitrogen (N) fertilizer and nitrification inhibitor dicyandiamide (DCD) on GHGs emissions. This study comprised four experimental treatments, including (i) control (CK), (ii) dicyandiamide, 20 mg kg−1 (DCD), (iii) nitrogen fertilizer, 300 mg kg−1 (N) and (iv) DCD + N. All experimental treatments were kept under flooded condition at the onset of the experiment, and then converted to 60% water filled pore space (WFPS). At flooding stage, N2O emissions were lower as compared to 60% WFPS. The highest cumulative N2O emission was 0.98 mg N2O–N kg−1 in N treated soil due to high substrates of mineral N contents, but lowest (0.009 mg N2O–N kg−1) in the DCD treatment. The highest cumulative CH4 emissions (80.54 mg CH4–C kg−1) were observed in the N treatment, while uptake of CH4 was observed in the DCD treatment. As flooded condition converted to 60% WFPS, CO2 emissions gradually increased in all experimental treatments, but the maximum cumulative CO2 emission was 477.44 mg kg−1 in the DCD + N treatment. The maximum dissolved organic carbon (DOC) contents were observed in N and DCD + N treatments with the values of 57.12 and 58.92 mg kg−1, respectively. Microbial biomass carbon (MBC) contents were higher at flooding while lower at transition phase, and increased at the initiation of 60% WFPS stage. However, MBC contents declined at the later stage of 60% WFPS. The maximum MBC contents were 202.12 and 192.41 mg kg−1 in N and DCD + N treatments, respectively. Results demonstrated that water regimes exerted a dramatic impact on C and N dynamics, subsequently GHGs, which were highly controlled by DCD at both flooding and 60% WFPS conditions.

Niche Differentiation Among Canonical Nitrifiers and N2O Reducers Is Linked to Varying Effects of Nitrification Inhibitors DCD and DMPP in Two Arable Soils.

The efficacy of nitrification inhibitors (NIs) dicyandiamide (DCD) and 3,4-dimethylpyrazole phosphate (DMPP) varies with soil types. Understanding the microbial mechanisms for this variation may lead to better modelling of NI efficacy and therefore on-farm adoption. This study addressed the response patterns of mineral nitrogen, nitrous oxide (N2O) emission, abundances of N-cycling functional guilds and soil microbiota characteristics, in relation to urea application with or without DCD or DMPP in two arable soils (an alkaline and an acid soil). The inhibition of nitrification rate and N2O emission by NI application occurred by suppressing ammonia-oxidizing bacteria (AOB) abundances and increasing the abundances of nosZI-N2O reducers; however, abundances of ammonia-oxidizing archaea (AOA) were also stimulated with NIs-added in these two arable soils. DMPP generally had stronger inhibition efficiency than DCD, and both NIs' addition decreased Nitrobacter, while increased Nitrospira abundance only in alkaline soil. N2O emissions were positively correlated with AOB and negatively correlated with nosZI in both soils and AOA only in acid soil. Moreover, N2O emissions were also positively correlated with nirK-type denitrifiers in alkaline soil, and clade A comammox in acid soil. Amendment with DCD or DMPP altered soil microbiota community structure, but had minor effect on community composition. These results highlight a crucial role of the niche differentiation among canonical ammonia oxidizers (AOA/AOB), Nitrobacter and Nitrospira, as well as nosZI- and nosZII-N2O reducers in determining the varying efficacies of DCD and DMPP in different arable soils.

Use of Symbiotic Fungi to Reduce the Phytotoxic Effect of DCD Nitrification Inhibitors in Lettuce

Nitrification inhibitors are commonly used to prevent nitrate leaching. However, the use of nitrification inhibitors is not free of side-effects. Some may be absorbed by the plant and cause phytotoxicity or even affect the food chain. Therefore, a solution that limits the absorption of nitrification inhibitors and its accumulation by the plant may mitigate health and environmental issues potentially associated with high levels of nitrification inhibitors. This solution may relay in the modulation of the plant’s metabolism through the interaction with specific fungal partners. This work tested the hypothesis that the symbiotic interaction between fungi and plant roots can reduce the destructive effects of the nitrification inhibitor Dicyandiamide (DCD) in plants by reducing the uptake of nitrification inhibitors. A greenhouse experiment was conducted, using a complete randomized block design, to test the effect of symbiotic fungi (plants inoculated with Piriformospora indica, Glomus etunicatum, and Glomus mosseae and noninoculated) on the phytotoxicity of DCD applied at four concentrations (0, 5, 50, and 100 mg kg−1 soil). Latuca sativa, cultivar Siyahoo, was selected for this experiment due to its economic value all over the world. The use of high DCD concentrations (100 mg kg−1 soil) affected the leaf chlorophyll content and plant growth in a manner that was significantly mitigated by the symbiosis of the plant with the fungal partner. These results highlight the benefits of using symbiotic fungal inoculants as plant protectors against the phytotoxic effects of DCD.

Effects of soil moisture on microbial processes of soil nitrogen gases production under anaerobic conditions

Gaseous nitrogen (N) emission [nitric oxide (NO), nitrous oxide (N2O), and nitrogen (N2)] is an important pathway of soil N loss. Nitrification and denitrification are the main processes of gaseous N production in soil. However, the contribution of heterotrophic nitrification, co-denitrification, and anammox to gaseous N production remains uncertain. In a laboratory soil incubation experiment, we used the 15N labelling and pairing technique, combining the nitrification inhibitor dicyandiamide (DCD), to quantify the contribution of different microbial processes to soil NO, N2O and N2 production under anaerobic conditions. The results showed that after 24 h anaerobic incubation, the highest total 15N recovery of three gases occurred at 65% water filled pore space (WFPS), accounting for 20.0% of total added 15N. Denitrification contributed 49.9%-94.1%, 29.0%-84.7%, and 58.2%-85.8% to the production of NO, N2O and N2 respectively, suggesting that denitrification was the predominant process of those three N gases emission. Heterotrophic nitrification was an important pathway of NO and N2O production, particularly at conditions with low soil water content (10% WFPS), with its contribution to those two N gases production being 50.1% and 42.8%, respectively. Co-denitrification contributed 10.6%-30.7% of N2O production. For N2 production, the total contribution of co-denitrification and anammox was 14.2%-41.8%. The role of co-denitrification can not be ignored for N2O and N2 production. Our results demonstrated that the 15N labelling and pairing technique is a promising tool to quantify the contribution of different microbial processes to gaseous N loss.