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.     

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.     

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.

     

Effect of urease inhibitors on nitrification in soil

Abstract. This paper describes experiments on the inhibitory effects of hydroquinone (HQ), phenylphos-phorodiamidate (PPDA) and N-butyl phosphorothioic triamide (NBPT) on the nitrification in soil. Incubations were carried out at 2/3 field capacity at 25°C of soil samples to which either ammonium or nitrite was added together with inhibitors. Addition of PPDA or NBPT did not influence the oxidation of ammonium. HQ. however, retarded the process significantly, and also the accumulation of nitrite. This was confirmed in experiments whereby added nitrite was followed. Some of the differences could be explained by changes in the soil pH. During incubation the evolution of the total mineral nitrogen was not importantly altered by addition of the inhibitors.     

Suppression of methane oxidation in aerobic soil by nitrogen fertilizers,nitrification inhibitors,and urease inhibitors

Concentrations of CH₄, a potent greenhouse gas, have been increasing in the atmosphere at the rate of 1% per year. The objective of these laboratory studies was to measure the effect of different forms of inorganic N and various N-transformation inhibitors on CH₄ oxidation in soil. NH ₄ ⁺ oxidation was also measured in the presence of the inhibitors to determine whether they had differential activity with respect to CH₄ and NH ₄ ⁺ oxidation. The addition of NH₄Cl at 25 g N g^-1 soil strongly inhibited (78–89%) CH₄ oxidation in the surface layer (0–15 cm) of a fine sandy loam and a sandy clay loam (native shortgrass prairie soils). The nitrification inhibitor nitrapyrin (5 g g^-1 soil) inhibited CH₄ oxidation as effectively as did NH₄Cl in the fine sandy loam (82–89%), but less effectively in the sandy clay loam (52–66%). Acetylene (5 mol mol^-1 in soil headspace) had a strong (76–100%) inhibitory effect on CH₄ consumption in both soils. The phosphoroamide (urease inhibitor) N-(n-butyl) thiophosphoric triamide (NBPT) showed strong inhibition of CH₄ consumption at 25 g g^-1 soil in the fine sandy loam (83%) in the sandy clay loam (60%), but NH ₄ ⁺ oxidation inhibition was weak in both soils (13–17%). The discovery that the urease inhibitor NBPT inhibits CH₄ oxidation was unexpected, and the mechanism involved is unknown.     

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.     

Persistence of effects of nitrification inhibitors added to soils

Abstract

The persistence of the effects of four nitrification inhibitors (2‐ethynylpyridine, nitrapyrin, etridiazole, 3‐methylpyrazole‐l‐carboxamide) on nitrification in soil was assessed by measuring the ability of two soils to nitrify NH₄ ⁺ [added as (NH₄)₂SO₄] after they had been treated with 5 μg inhibitor g^‐1 soil and incubated at 10, 20, or 30°C for 0, 21, 42, 84, 126, or 168 days. The soils used differed markedly in organic‐matter content (1.2 and 4.2% organic C). The data obtained showed that the persistence of the effects of the inhibitors studied decreased markedly with increase in soil temperature from 10 to 30°C and that, whereas the initial inhibitory effects of the test compounds on nitrification were greatest with the soil having the lower organic‐matter content, the persistence of their effects at 20 or 30°C was greatest with the soil having the higher organic‐matter content. The inhibitory effects of 2‐ethynylpyridine and etridiazole on nitrification were considerably more persistent than those of nitrapyrin or 3‐methylpyrazole‐l‐carboxamide and were significant even after incubation of inhibitor‐treated soil at 20°C for 168 days.     

不同盐渍化土壤中微生物对氮肥的响应

【目的】 研究河套灌区土壤不同盐渍化程度下土壤微生物对氮肥的响应机理,为河套灌区盐渍化土壤中确立适宜的氮肥施用量提供理论依据。 【方法】 2015—2016年在内蒙古磴口县坝楞示范基地进行了两年田间试验,供试土壤为粉沙壤土,供试作物为玉米。设置轻度 (0.77～1.24 mS/cm) 和中度 (1.24～1.77 mS/cm) 盐渍化土壤为主区,副区为施氮水平,共设4个施氮水平为N 0、135、270、405 kg/hm2,研究轻、中度盐渍化土壤下不同施氮水平对土壤微生物的影响,探寻土壤盐渍化和氮肥对微生物的交互作用。 【结果】 土壤盐分随着施氮量的增加而增加;细菌、真菌、放线菌数量和微生物量碳、氮含量均随着施氮量的增加呈现出先增加后降低的趋势,在N 270 kg/hm2处理微生物数量和生物量均达到最高值。回归分析表明,土壤微生物与氮、土壤盐分之间呈现出极显著的二元二次非线性回归关系,由回归方程各系数可知,在盐渍化土壤中,单独施用氮肥均可以增加土壤微生物,而土壤盐分增加,土壤微生物减少;在轻度盐渍化土壤中,土壤盐分和施用氮肥对微生物具有正效应,即共同促进了土壤微生物的增加,而在中度盐渍化土壤中,土壤盐分和施用氮肥对微生物具有负效应,即抑制土壤微生物的增加。 【结论】 土壤盐渍化程度越高,土壤中微生物数量和生物量越少,施氮量为N 270 kg/hm2时微生物数量和生物量均达到最大值;土壤微生物与氮、土壤盐分之间呈现出极显著的二元二次非线性回归关系,在轻度盐渍化土壤中,土壤盐分和施用氮肥共同促进土壤微生物的增加,而在中度盐渍化土壤中,土壤盐分和施用氮肥抑制了土壤微生物的生长和繁殖。      

棕壤线虫对尿素配施尿酶抑制剂和硝化抑制剂的响应研究

Effects of urea amended with urease and nitrification inhibitors on soil nematode communities were studied in a Hapli- Udic Argosol （Cambisol, FAO） in Liaoning Province of Northeast China. A completely random design with four treatments, i.e., conventional urea （CU）, slow-release urea amended with a liquid urease inhibitor （SRU1）, SRU1 ＋nitrification inhibitor dicyandiamide （SRU2）, and SRU1 ＋ nitrification inhibitor 3,5-dimethylpyrazole （SRU3） and four replicates were applied. Thirty-nine genera of nematodes were identified, with Cephalobus and Aphelenchus being dominant; and in all treatments, the dominant trophic group was bacterivores. In addition, during the growth period of spring wheat （Triticum aestivum L.）, soil urease activity was lower in SRUs than in CU. The numbers of total nematodes and bacterivores at wheat heading and ripening stages, and omnivores-predators at ripening stage were higher in SUR3 than in CU, SRU1 and SRU2 （P 〈 0.05）.     

Sorption-desorption of indaziflam in selected agricultural soils

Indaziflam, a new alkylazine herbicide that inhibits cellulose biosynthesis, is under current development for soil applications in perennial crops and nonagricultural areas. Sorption and desorption of indaziflam in six soils from Brazil and three soils from the United States, with different physical chemical properties, were investigated using the batch equilibration method. Sorption kinetics demonstrated that soil-solution equilibrium was attained in <24 h. The Freundlich equation described the sorption behavior of the herbicide for all soils (R(2) > 0.99). K(f) values of the Brazilian oxisols ranged from 4.66 to 29.3, and 1/n values were ≥ 0.95. Sorption was positively correlated to %OC and clay contents. U.S. mollisol K(f) values ranged from 6.62 to 14.3; 1/n values for sorption were ≥ 0.92. K(f) values from mollisols were also positively correlated with %OC. These results suggest that indaziflam potential mobility, based solely on its sorption coefficients, would range from moderate to low in soil. Desorption was hysteretic on all soils, further decreasing its potential mobility for offsite transport.     

脲酶抑制剂与硝化抑制剂对稻田氨挥发的影响

采用密闭室间歇通气法和15N标记技术研究了尿素施入稻田后氨挥发损失特征以及脲酶抑制剂(N-丁基硫代磷酰三胺,NBPT)和硝化抑制剂(3, 4-二甲基吡唑磷酸盐,DMPP)对稻田氨挥发损失的影响。结果表明,稻田施用尿素后第4天氨挥发速率达到峰值,氨挥发损失主要发生在施肥后21天内。与单施尿素处理相比,添加NBPT处理的氨挥发速率峰值降低27.04%,累积氨挥发损失量降低21.65%;NBPT与DMPP配施时,氨挥发速率峰值降低12.95%,累积氨挥发损失量降低13.58%;而添加DMPP时,氨挥发速率峰值增加23.61%,累积氨挥发损失量与单施尿素的差异不显著。相关性分析表明,地表水中铵态氮浓度和pH值与氨挥发速率均达极显著正相关,说明二者是影响氨挥发速率的主要因素,而气温、 地温和水温与氨挥发速率的相关性不显著。与单施尿素相比,添加脲酶抑制剂可显著增加稻谷产量。脲酶抑制剂与硝化抑制剂配合施用可更有效地提高氮肥的回收率。综合降低氨挥发、 提高水稻产量及地上部氮肥回收率的效果,添加脲酶抑制剂以及脲酶抑制剂与硝化抑制剂配施的两个处理效果较为理想,硝化抑制剂不宜单独添加。     

Effect of urease and nitrification inhibitors on ammonia volatilization and abundance of N‐cycling genes in an agricultural soil

The effect of the combined application of urease and nitrification inhibitors on ammonia volatilization and the abundance of nitrifier and denitrifier communities is largely unknown. Here, in a mesocosm experiment, ammonia volatilization was monitored in an agricultural soil treated with urea and either or both of the urease inhibitor N‐(n‐butyl) thiophosphoric triamide (NBPT) and the nitrification inhibitor 3,4‐dimethylpyrazole phosphate (DMPP), with 50% and 80% water‐filled pore space (WFPS). The effect of the treatments on the abundance of bacteria and archaea was estimated by quantitative PCR (qPCR) amplification of their respective 16S rRNA gene, that of nitrifiers using amoA genes, and that of denitrifiers by qPCR of the norB and nosZI denitrification genes. After application of urea, N losses due to NH₃ volatilization accounted for 23.0% and 9.2% at 50% and 80% WFPS, respectively. NBPT reduced NH₃ volatilization to 2.0% and 2.4%, whereas DMPP increased N losses by up to 36.8% and 26.0% at 50% and 80% WFPS, respectively. The combined application of NBPT and DMPP also increased NH₃ emissions, albeit to a lesser extent than DMPP alone. As compared with unfertilized control soil, both at 50% and 80% WFPS, NBPT strongly affected the abundance of bacteria and archaea, but not that of nitrifiers, and decreased that of denitrifiers at 80% WFPS. Regardless of moisture conditions, treatment with DMPP increased the abundance of denitrifiers. DMPP, both in single and in combined application with NBPT, increased the abundance of nitrification and denitrification genes.     

Influence of water potential on nitrification and structure of nitrifying bacterial communities in semiarid soils

The objective of this study was to examine the relationship between soil water potential, nitrifier community structure and nitrification activity in semiarid soils. Soils were collected after a 5-month dry period (end of summer) and subsequently rewetted to specific water potentials and incubated for 7 days prior to analysis of nitrification activity and nitrifier community structure. The approach used in this study targeted a 491bp segment of the amoA gene which encodes the active site of the ammonia monooxygenase enzyme, which is the key enzyme for all aerobic ammonia oxidisers. amoA serves as a useful target for environmental studies since it is both specific and universal for all ammonia oxidisers and reflects the phylogeny of the ammonia oxidisers. Our results suggest that in semiarid soils water potential plays a key role in determining the structure of ammonia oxidising bacteria (AOB), and that additionally AOB community structure is correlated to potential nitrification rate in these soils.     

脲酶/硝化抑制剂对尿素氮在白浆土中转化的影响

采用室内恒温培养方法,研究了脲酶抑制剂（NBPT）、硝化抑制剂（DMPP）及其协同对尿素氮在三江平原白浆土中转化作用效果。研究表明,在白浆土中NBPT有效作用时间小于13 d,作用时间较在棕壤和黑土中短;对土壤中铵态氮、硝态氮及表观硝化率影响与普通尿素基本一致。NBPT与DMPP组合缓释尿素施入4-7 d,能够有效抑制脲酶活性,减缓尿素水解;只添加DMPP与添加NBPT与DMPP协同作用对抑制铵态氮硝化作用效果相同,二者能保持土壤中NH4+-N高含量时间超过80 d。DMPP作用时间可达80 d以上,能有效抑制NH4+-N向NO3--N的转化;在第80 d,土壤中仍有54.58%～56.85%的氮以铵态氮形式存在,表观硝化率只有50%左右。DMPP抑制硝化作用效果十分显著,因此,在白浆土中施用添加NBPT缓释尿素、DMPP缓释尿素、NBPT与DMPP缓释尿素时,应首选添加1%DMPP的缓释尿素肥料。     

Effects of afforestation on ammonification and nitrification rates in former agricultural soils

Abstract. The effects of afforestation on potential nitrification, nitrification and ammonification rates were studied at an experimental site in NE Scotland 4½ years after afforestation of former arable land. The site had been planted with three tree species (Sitka spruce, sycamore and hybrid larch) at three different planting densities, with half the plots treated with inorganic NPK fertilizer. Laboratory measurements of potential nitrification, nitrification and ammonification rates, measured using a perfusion system, were compared between the unforested control and combinations of the various treatments. Differences in soil pH and soil moisture content were also investigated.
Potential nitrification rates measured in plantation soils were significantly lower than in the unplanted control soil. Nitrification and ammonification rates were also consistently lower, although these differences were only significant in a few of the treatments. Soils planted with a normal tree density had a tendency to show higher nitrification rates compared to soils planted with a high tree density.
The results suggest that afforestation of former agricultural soils may cause changes in important parts of the soil N cycle soon after planting. At this early stage in the life of the plantation this appears to be unrelated to changes in soil pH or moisture content, even though soils beneath the trees are drier. The apparent change may be the result of differences in the soil microbial community associated with the type of organic matter substrate present in the unplanted and planted soils.     

Nitrous oxide production by denitrification and nitrification in temperate forest, grassland and agricultural soils

Nitrous oxide is produced in soils by biological denitrification and nitrification. To improve the fundamental understanding of the processes leading to N₂O fluxes from soils, the production of N₂O from denitrification and nitrification in spruce forest, beech forest, riparian grassland, coastal grassland and an agricultural field were studied. Samples were taken at a high and a low position along a topographic gradient in each site in the spring and autumn when the largest N₂O fluxes were expected. They were incubated after being amended with N, and C₂H₂ was used as biological inhibitor to distinguish nitrification and denitrification. The N₂O production in the low landscape position varied between 32 and 121 ng N cm^?3 h^?1 in the riparian grassland. 9 and 26 ng N cm^?3 h^?1 in the coastal grassland, and 135 and 195 ng N cm^?3 h^?1 in the agricultural field which was 10–100 times more than in the high positions where rates ranged between 3 and 5 ng N cm^?3 h^?1, 0.3 and 0.4 ng N cm^?3 h^?1, and 7 and 10 ng N cm^?3 h^?1, respectively. These differences almost certainly arose because the soil in the low positions was wetter and contained more organic matter. In the two forests N₂O production was less than 1 ng N cm^?3 h^?1, strongly inhibited by O₂, and not influenced by landscape position. Nitrification contributed to more than 60% of total N₂O production in the riparian grassland. In the agricultural field nitrification produced 13–74% of the total N₂O in the low position, and 10–88% in the high position. Denitrification was the dominant source of N₂O in the coastal grassland except at the low position in the autumn where nitrification produced 60% of the total N₂O. In the two forests where the soil had small nitrification potentials denitrification was the only source of N₂O. In the other sites nitrification and denitrification potentials were large and of identical magnitude. The results emphasize the need to separate nitrification and denitrification at the process level and to recognize topography at the field scale when modelling N₂O effluxes from soil.     

硝化/脲酶抑制剂对秋马铃薯植株及土壤氮素利用的影响

通过添加硝化/脲酶抑制剂达到秋马铃薯生产中减氮增效的目的,为制定秋马铃薯的农田氮素管理措施及节肥增效策略提供依据.采用随机区组试验设计,设置30、60、90、120 kg/hm24个施氮水平与硝化/脲酶抑制剂N-丁基硫代磷酰三胺(NBPT)、双氰胺(DCD)、2-氯-6-三氯甲基吡啶(CP)、未添加硝化/脲酶抑制剂(C...     

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.