Effects of urease inhibitors on nitrification in soils

The effects of 10 urease inhibitors on nitrification in soils were studied by determining the effects of 10 and 50 parts/10⁶ (soil basis) of each inhibitor on the amounts of nitrate and nitrite produced when soils treated with ammonium sulfate (200 μg of ammonium N/g of soil) were incubated (30°C) under aerobic conditions for 14 days. The urease inhibitors used (catechol. hydroquinone, p-benzoquinone, 2,3-dimethyl-p-benzoquinone, 2,5-dimethyl-p-benzoquinone. 2,6-dimethyl-p-benzoquinone. 2,5-dichloro-p-benzoquinone, 2,6-dichloro-p-benzoquinone. sodium p-chloromercuribenzoate, and phenylmercuric acetate) were those found most effective in previous work to evaluate more than 130 compounds as soil urease inhibitors. Their effects on nitrification were compared with those of three compounds patented as soil nitrification inhibitors (N-Serve. AM. and ST).Most of the urease inhibitors studied had little effect on nitrification when applied at the rate of 10 μg/g of soil. but had marked inhibitory effects when applied at the rate of 50 μg/g of soil. None inhibited nitrification as effectively as N-Serve. but phenylmercuric acetate inhibited nitrification more effectively than did AM or ST when applied at the rate of 10 μg/g of soil. Phenylmercuric acetate, 2,5-dimethyl-p-benzoquinone, and 2,6-dimethyl-p-benzoquinone had very marked effects on nitrification when applied at the rate of 50 μg/g of soil.     

Heterotrophic nitrification by sodium chloride-tolerant fungi in soils added with sodium chloride

Abstract

Recently, fungi with the ability of heterotrophic nitrification have been isolated from acid forest soils (Lang and Jagnow 1986; Stroo et al. 1986). It is suggested that under unsuitable conditions for autotrophic nitrification, heterotrophic processes for NO₂ ^- and NO_{3 -} production could be advantageous.     

Effects of nitrification inhibitors on transformations of urea nitrogen in soils

The effects of three patented nitrification inhibitors on transformations of urea N in soils were studied by determining the effects of these compounds (10 μg/g of soil) on urea hydrolysis, ammonia volatilization. and production of ammonium, nitrite, and nitrate in soils incubated under aerobic conditions (30°C, 60% WHC) after treatment with urea (400 μg of urea N/g of soil). The inhibitors used (N-Serve, ATC, and CL-1580) had little, if any, effect on urea hydrolysis, but they retarded nitrification of the ammonium formed by urea hydrolysis and increased gaseous loss of urea N as ammonia. They also decreased the amount of (urea + exchangeable ammonium + nitrite + nitrate) — N found in urea-treated soils after various times.Two of the soils used accumulated substantial amounts of nitrite(> 160 μg of nitrite N/g of soil) when incubated under aerobic conditions after treatment with urea. Addition of nitrification inhibitors to these soils eliminated or substantially reduced nitrite accumulation and greatly retarded nitrate formation, but had little, if any, effect on the recovery of urea N as (urea + exchangeable ammonium + nitrite + nitrate + ammonia) — N after various times. This finding and other observations reported indicate that the “nitrogen deficits” observed in studies of urea N transformations in soils may not largely be due to gaseous loss of urea N through chemodenitrification and are at least partly due to volatilization and fixation of the ammonium formed by urea hydrolysis in soils. The work reported also indicates that N-Serve and other nitrification inhibitors may prove useful for reduction of the nitrite toxicity problems associated with the use of urea as a fertilizer but that application of such inhibitors in conjunction with fertilizer urea, when surface applied, may promote gaseous loss of urea N as ammonia.     

Release of mineral N from soils: Influence of inhibitors of nitrification

Nitrification inhibitors (N-Serve, ATC, and CS₂) were added to soils without N fertilizers. While the amount of nitrification of NH₄⁺-N was reduced, so was the amount of ammonification of soil N. This effect was greater with ATC and CS₂ than with N-Serve. In three field experiments, the application in the fall of ATC at 22 kg ha^?1 mixed into the soil reduced the loss of soil mineral N in early spring. Apparently, the inhibition suppressed both ammonification and nitrification of soil N during the winter, and consequently there was less NO^?₃ in soil when the wet period occurred in the spring.     

Effects of urease and nitrification inhibitors added to urea on nitrous oxide emissions from a loess soil

Urea fertilizer‐induced N₂O emissions from soils might be reduced by the addition of urease and nitrification inhibitors. Here, we investigated the effect of urea granule (2–3 mm) added with a new urease inhibitor, a nitrification inhibitor, and with a combined urease inhibitor and nitrification inhibitor on N₂O emissions. For comparison, the urea granules supplied with or without inhibitors were also used to prepare corresponding supergranules. The pot experiments without vegetation were conducted with a loess soil at (20 ± 2)°C and 67% water‐filled pore space. Urea was added at a dose of 86 kg N ha^–1 by surface application, by soil mixing of prills (<1 mm) and granules, and by point‐placement of supergranules (10 mm) at 5 cm soil depth. A second experiment was conducted with spring wheat grown for 70 d in a greenhouse. The second experiment included the application of urea prills and granules mixed with soil, the point‐placement of supergranules and the addition of the urease inhibitor, and the combined urease plus nitrification inhibitors at 88 kg N ha^–1. In both experiments, maximum emissions of N₂O appeared within 2 weeks after fertilization. In the pot experiments, N₂O emissions after surface application of urea were less (0.45% to 0.48% of total fertilization) than from the application followed by mixing of the soil (0.54% to 1.14%). The N₂O emissions from the point‐placed‐supergranule treatment amounted to 0.64% of total fertilization. In the pot experiment, the addition of the combined urease plus nitrification inhibitors, nitrification inhibitor, and urease inhibitor reduced N₂O emissions by 79% to 87%, 81% to 83%, and 15% to 46%, respectively, at any size of urea application. Also, the N₂O emissions from the surface application of the urease‐inhibitor treatment exceeded those of the granules mixed with soil and the point‐placed‐supergranule treatments receiving no inhibitors by 32% to 40%. In the wheat growth experiment, the N₂O losses were generally smaller, ranging from 0.16% to 0.27% of the total fertilization, than in the pot experiment, and the application of the urease inhibitor and the combined urease plus nitrification inhibitors decreased N₂O emissions by 23% to 59%. The point‐placed urea supergranule without inhibitors delayed N₂O emissions up to 7 weeks but resulted in slightly higher emissions than application of the urease inhibitor and the urease plus nitrification inhibitors under cropped conditions. Our results imply that the application of urea fertilizer added with the combined urease and nitrification inhibitors can substantially reduce N₂O emissions.     

Growth of comammox Nitrospira is inhibited by nitrification inhibitors in agricultural soils

Purpose

The discovery of comammox Nitrospira being capable of complete oxidising ammonia to nitrate radically challenged the conventional concept of two-step nitrification. However, the response of comammox Nitrospira to nitrification inhibitors (NIs) and their role in soil nitrification remain largely unknown, which has hindered our ability to predict the efficiency of NIs in agroecosystems.

Materials and methods

We evaluated the effect of four NIs, 2-chloro-6-(trichloromethyl) pyridine (nitrapyrin), 3,4-dimethylpyrazole phosphate (DMPP), allylthiourea (ATU) and dicyandiamide (DCD) on the growth of comammox Nitrospira, ammonia-oxidising archaea (AOA) and ammonia-oxidising bacteria (AOB) in two pasture and arable soils.

Results and discussion

The amendment of nitrogen fertiliser significantly increased soil nitrate concentrations over time, indicating a sustaining nitrification activity in both soils. The addition of all the four NIs effectively reduced the production of nitrate in both soils, but to varying degrees during incubation. The abundances of comammox Nitrospira clade A were significantly increased by addition of nitrogen fertilisers and significantly impeded by the four NIs in the pasture soil, but their abundances were only remarkably hindered by nitrapyrin in the arable soil. All the four NIs obviously inhibited the AOB abundances in both soils. Except for DMPP, the other three NIs effectively suppressed the AOA abundances in both soils.

Conclusions

We provided new evidence that growth of comammox Nitrospira clade A can be stimulated by nitrogen fertilisers and inhibited by various nitrification inhibitors, suggesting their potential role in nitrification of agricultural soils.

     

Dicyandiamide effects on nitrification and total inorganic soil nitrogen in sandy soils

Abstract

Inhibition of nitrification in soil results in a decreased ratio of nitrate‐nitrogen (NO₃‐N) to ammonium‐nitrogen (NH₄‐N). If the conditions for NO₃‐N loss by leaching or denitrification exist, nitrification inhibitors should increase concentrations of total inorganic soil nitrogen (N) (TISN) (NH₄‐N + NO₃‐N). This can then result in plants taking up more N and developing more crop yield or biomass. This study examined whether inhibition of nitrification by dicyandiamide (DCD) would result in increased concentrations of TISN under field conditions. The effects of DCD on soil N were evaluated in hyperthermic sandy soils planted to potato (Solanum tuberosum L., cv. Atlantic). Treatments were factorial combinations of N as ammonium nitrate (NH₄NO₃) at 67, 134, and 202 kg N ha^‐1 and DCD at 0, 5.6, and 11.2 kg DCD ha^‐1. Soil NH₄‐N, NO₃‐N, and TISN concentrations were determined for up to five potato growth stages at two locations for two years for a total of 16 determinations (cases), i.e., four were not determined. The N form ratio [NO₃‐N/(NH₄‐N + NO₃‐N] x 100 was decreased in 10 of 16 cases, indicating that nitrification was inhibited by DCD. With two of these 10 cases, TISN concentration increased, but with four others, TISN concentration decreased with at least one N rate. With four of these 10 cases, inhibition of nitrification had no effect on TISN concentration. Under the conditions of these field studies, DCD inhibited nitrification more often than not. Inhibition of nitrification was, however, more likely to reduce TISN concentration than to increase it. This may have been due to DCD effects on immobization of applied NH₄‐N.     

Responses of ureolytic and nitrifying microbes to urease and nitrification inhibitors in selected agricultural soils in Victoria,Australia

Journal of Soils and Sediments - Urease inhibitors (UIs) such as N-(n-butyl)thiophosphoric triamide (NBPT) and nitrification inhibitors (NIs) such as 3,4-dimethylpyrazole phosphate (DMPP) have been...     

Persistence of the inhibitory effects of phosphoroamides on urea hydrolysis in soils

Abstract

The persistence of the inhibitory effects of three phosphoroamides [N‐(n‐butyl) thiophosphoric triamide (NBPT), phenylphosphorodiamidate (PPD), and thiophosphoryl triamide (TPT)] on urea hydrolysis in soils was assessed by measuring the ability of four soils to hydrolyze urea after they had been treated with 5 μg phosphoroamide/g soil and incubated at 15°C or 30°C for 0, 3, 7, 14, or 28 days. The soils used differed markedly in pH, texture, and organic‐matter content. The data obtained showed that the persistence of the effects of the phosphoroamides studied decreased with increase in soil temperature from 15°C to 30°C and that whereas the effect of PPD decreased with increase in the time of incubation, the effects of NBPT and TPT sometimes increased before decreasing with increased time of incubation. These observations are in harmony with the recent findings that PPD is a potent inhibitor of urease activity, but decomposes in soils with formation of phenol, which is a relatively weak inhibitor of urease activity, whereas NBPT and TPT do not inhibit urease activity but decompose in soil with formation of their oxon analogs, which are potent inhibitors of urease activity. The inhibitory effect of NBPT on urea hydrolysis was considerably more persistent than that of PPD or TPT and was significant even after incubation of NBPT‐treated soil at 15°C or 30°C for 28 days.     

Modes of action of nitrification inhibitors

 In recent years, substantial progress has been made towards understanding the modes of action for the specific inhibition of autotrophic NH₃ oxidation. This has included demonstrating that NH₃ monooxygenase (AMO) has a broad substrate range for catalytic oxidation, and the inhibitory effects of many compounds are due to competition for the active site. Other compounds, such as acetylenes, are oxidized by the normal catalytic cycle of AMO to highly reactive products which covalently bind the enzyme causing irreversible inhibition. Substantial evidence has shown the important role of Cu in the activity of AMO, and indicated that a large class of compounds containing thiono-S inhibit AMO activity by binding with Cu within the active site. Heterocyclic N compounds form another important class of nitrification inhibitors with little known about their mode of action, although evidence suggests that their inhibitory influence is closely related to the presence of ring N. Received: 21 July 1998     

A hot-spot approach applied to nitrification in tropical acid soils

Several authors have reported that nitrification in acid soils may be restricted to microsites having a more favorable pH. The aim of this study was to propose a conceptual model of the functioning of nitrification in hot-spots, and to test it with the experimental data obtained in laboratory conditions using twelve tropical unamended and -amended soils with a wide range of pH (from 4.2 to 6.9). Nitrification was also measured in two selected soils where the pH was adjusted from 3.5 to 6.2. The model characterizes the relationship between the nitrification rates in unamended and -amended soils as a function of pH. It is based upon the assumption that nitrification of the coming from N mineralization occurs in the hot-spot (RN_h), and the nitrification of the added occurs in the hot-spot and also in its adjacent surrounding region (RN_s). The experimental design was chosen to be able to estimate both nitrification rates. Soil acidity limited nitrification more in -amended soils than in unamended ones. From our approach, this is due to less favorable conditions for nitrification in the region surrounding the hot-spot. The effect of self-induced acidity on nitrification was not noticeable neither in unamended nor in -amended treatments. The model described well three observations made in the experiments: (i) the minimum pH for nitrification to occur was lower for RN_h (pH<4.2) than for RN_s (pH<4.7), (ii) the RN_h/RN_s ratio increased with the decrease of pH (from 1.5 at pH 6 to 8.5 at pH 4), and (iii) for a given pH, the RN_h/RN_s ratio increased with the decrease of the initial pH of the soil. Among the soil parameters determined in this study (i.e. exchangeable Al, EDTA-extractable Cu and Zn, total C and N), only pH was related to nitrification. However, for a given pH, nitrification varied 3-fold among soils, depending upon their initial pH. This suggests that soil pH as determined on bulk soil is not suitable to predict nitrification in each individual soil, because it is not representative of the acidity level within the hot-spot.     

Effect of temperature on nitrification in soils

Nitrification plays an important role in nitrogen transformation in soils. As a practical problem the fact is noteworthy that by nitrification, the loss of nitrogen from soils is caused at higher temperatures in the summer. On the contrary, it is quite slow at the lower temperatures of early spring. Recently much Work has been done on the effect of temperature on nitrification in soils. It was noticed that its effect is not the same according to the difference of soils. Soils which have high nitrifying activities at ordinary temperatures (around 25°C) show fairly good nitrification even at lower temperatures (1, 2, 5). However, in nitrification by usual experimental methods, the amount of nitrate produced can be only estimated while nitrifying organisms are increasing or decreasing in soil. As the growing Process of microbes is extremely influenced by both the properties of the soil and the cultural conditions, it is necessary to examine the effect of temperature on the growing process of microbes rather than biochemical nitrification by a unit cell. In usual methods nitrification can hardly be treated in these separate aspects. The author has succeeded in obtaining soils in which nitrifying organisms increase to the maximum limit, and no increase can be expected in number of these organisms. By using such samples, experiments were made concerning the effect of temperature on nitrification by a unit cell and its direct effect on the growth of organisms. Furthermore, the author prepared favorable cultural conditions for various kinds of soils by the “washing cultivation method” (6), and examined the effect of temperature on the increasing process of nitrifying ability.     

两种农田土壤中的氨氧化细菌和氨氧化古菌对硝化抑制剂DCD和DMPP的响应-盆栽试验

Taking two important agricultural soils with different pH, brown soil （Hap-Udic Luvisol） and cinnamon soil （Hap-Ustic Luvisol）, from Northeast China, a pot culture experiment with spring maize （Zea mays L.） was conducted to study the dynamic changes in the abundance and diversity of soil ammonia-oxidizing bacteria （AOB） and ammonia-oxidizing archaea （AOA） populations during maize growth period in response to the additions of nitrification inhibitors dicyandiamide （DCD） and 3,4-dimethylpyrazole phosphate （DMPP） by the methods of real-time polymerase chain reaction （PCR） assay, PCR-denaturing gradient gel electrophoresis （DGGE）, and construction of clone library targeting the amoA gene. Four treatments were established, i.e., no urea （control）, urea, urea plus DCD, and urea plus DMPP. Both DCD and DMPP inhibited growth of AOB significantly, compared to applying urea alone. Soil bacterial amoA gene copies had a significant positive linear correlation with soil nitrate content, but soil archaeal amoA gene copies did not. In both soils, all AOB sequences fell within Nitrosospira or Nitrosospira-like groups, and all AOA sequences belonged to group 1.1b crenaxchaea. With the application of DCD or DMPP, community composition of AOB and AOA in the two soils had less change except that the AOB community composition in Hap-Udic Luvisol changed at the last two growth stages of maize under the application of DCD. AOB rather than AOA likely dominated soil ammonia oxidation in these two agricultural soils.     

农田土壤中氮同位素分级与硝化能力的关系研究

A laboratory-based aerobic incubation was conducted to investigate nitrogen(N) isotopic fractionation related to nitrification in five agricultural soils after application of ammonium sulfate((NH4)2SO4). The soil samples were collected from a subtropical barren land soil derived from granite(RGB),three subtropical upland soils derived from granite(RQU),Quaternary red earth(RGU),Quaternary Xiashu loess(YQU) and a temperate upland soil generated from alluvial deposit(FAU). The five soils varied in nitrification potential,being in the order of FAU YQU RGU RQU RGB. Significant N isotopic fractionation accompanied nitrification of NH+4. δ15N values of NH+4 increased with enhanced nitrification over time in the four upland soils with NH+4 addition,while those of NO-3 decreased consistently to the minimum and thereafter increased. δ15N values of NH+4 showed a significantly negative linear relationship with NH+4-N concentration,but a positive linear relationship with NO-3-N concentration. The apparent isotopic fractionation factor calculated based on the loss of NH+4 was 1.036 for RQU,1.022 for RGU,1.016 for YQU,and 1.020 for FAU,respectively. Zero- and first-order reaction kinetics seemed to have their limitations in describing the nitrification process affected by NH+4 input in the studied soils. In contrast,N kinetic isotope fractionation was closely related to the nitrifying activity,and might serve as an alternative tool for estimating the nitrification capacity of agricultural soils.     

Potency of nitrification inhibitors following their repeated application to soil

Summary Nitrification inhibitors were applied to a field experiment on loamy sand soil each autumn for 4 years, immediately prior to sowing winter cereals. Laboratory experiments demonstrated that repeated application of the inhibitor dicyandiamide (DCD) to a soil had little effect either on the rate of DCD decomposition or the ability of DCD to inhibit nitrification. Repeated field application of the inhibitors DCD, nitrapyrin or etridiazole resulted in increased sensitivity of ammonium-oxidizing bacteria to nitrapyrin or etridiazole, but not to DCD. The rate of decomposition of etridiazole was unaffected by four annual applications of this inhibitor, but decomposition of nitrapyrin was somewhat slower in soil that had received nitrapyrin annually for 4 years than in soil that had never been treated with an inhibitor.     

Evolution of organic matter added to soils under cultivation conditions

The development of an organic matter (OM) based on mixed sheep manure and peat, when it was incorporated into soils as fertilizer, was studied. The experiment was carried out in soils under almond tree culture, with drip irrigation and non irrigation regimes. Two doses, 10 and 4.5 kg tree^–1, were assayed. Changes in the humic acid fraction one year after incorporation into soils showed oxidation and enrichment in condensed structures, as observed by an increase of the O*:H* ratio and a decrease of the H*:C* ratio, and also by FTIR spectra. The oxidative process was more significant in the coarser textured and also in the non‐irrigated soil. The evolution of the ratios C_ext:C_ox and C_HA:C_FA throughout the culture cycle was followed by sampling and chemical analysis of different forms of organic carbon. Evolution of C_ext:C_ox showed a uniform humification state in the irrigated soil, and a significant decrease in the non‐irrigated soil, at the beginning of the experiment. Curves of C_HA:C_FA evolution showed changes attributed to mineralization or drainage of the fulvic acids fraction, giving a maximum in spring in both soils and a final increase at the end of the cycle by drainage only in the irrigated soil.     

Hydrolysis of organic and inorganic phosphorus compounds added to soils

The rates of hydrolysis of seven organic and two inorganic phosphorus compounds applied to soils at a rate of 500 ppm P and incubated at 20°C for various times under aerobic and waterlogged conditions were studied. Monomethyl phosphate, β-glycerophosphate, and α-D-glucose-1-phosphate were hydrolyzed at similar rates in the three soils used, but the rates were somewhat faster under aerobic than under waterlogged conditions. Organic P compounds in which two hydrogens of the orthophosphoric acid are replaced (e.g., diphenyl phosphate) were hydrolyzed at slower rates than those in which one hydrogen is replaced (e.g., phenyl phosphate). The rate of hydrolysis of diphenyl phosphate was lower than that of bis-p-nitrophenyl phosphate. Of the two inorganic P compounds studied, ammonium tetrametaphosphimate did not hydrolyze in soil, and the rate of hydrolysis of phosphonitrilic hexaamide was very small (6–13% hydrolyzed in 7 days) compared with those of the organic phosphates (30–98%).     

Some furfural derivatives as nitrification inhibitors

Three series of furfural derivatives, namely N-O-furfural oxime ethers, furfural Schiff bases (furfurylidene anilines), and furfural chalcones, have been synthesized and evaluated for nitrification inhibition activity in laboratory incubation studies in typic Ustocrept soil. Furfural oxime ethers and furfural Schiff bases showed potential activity, but furfural chalcones were only mildly active. N-O-ethyl furfural oxime among the oxime ethers, and furfurylidine-4-chloroaniline among the furfural Schiff bases, performed the best. These two compounds showed more than 50% nitrification inhibition on the 45th day at 5% dose as compared to 73% inhibition by nitrapyrin. Activity of furfural oxime ethers decreased with an increase in carbon atoms in the N-O-alkyl side chain. Introduction of a chlorine atom in the phenyl ring of furfurylidene anilines increased the persistence of their activity. N-O-Ethyl furfural oxime and furfurylidine-4-chloroaniline coated urea performed at par with their application in solution form. Ethyl and N-O-isopropyl oxime, as well as chloro- and nitro- substituted Schiff bases, did not reveal any phytotoxicity (adverse effect on germination) on chickpea seeds (Cicer arietinum) even at the highest dose (40 ppm, soil basis).     

Relationships between stability of Cu complexes and nitrification inhibition effects of corresponding ligands in soils

In order to investigate the relationships between the stability of Cu complexes and the nitrification inhibition effects of corresponding ligands, the effects of 22 kinds of Cu (II) ligands and eight kinds of Cu (I) ligands were evaluated in three types of soil on nitrification potentials (NPs) under short-term incubation (5 h) and nitrification inhibitory rates (NIRs) under long-term incubation (14 d). The results showed that the inhibitory effect of Cu (II) or Cu (I) ligands on nitrification exhibited significant linear relationships (P < 0.01) with the logarithms of the first stability constants (lgK₁) and the total stability constants (lgK_f) of corresponding complexes. Furthermore, lgK_f of Cu (II) complexes exhibited a greater linear correlation than the corresponding lgK₁ with the nitrification inhibition effects of the corresponding ligands; lgK_f of Cu (I) complexes exhibited a steeper linear relationship than that of Cu (II) complexes with NPs or NIRs, and hence, nitrification inhibition effects of Cu (I) ligands almost doubled that of Cu (II) ligands when the lgK_f values of Cu (I) and Cu (II) complexes have the same changes.