土壤脲酶抑制剂正丁基硫代磷酰三胺的作用基团研究

土壤脲酶抑制剂正丁基硫代磷酰三胺(nBPT)是抑制土壤中尿素水解的最有效的化合物之一。分析大连工业大学合成的土壤脲酶抑制剂nBPT抑制脲酶活性的影响因素及作用机理,结果表明:在50°C,pH=5.91时,nBPT的抑制活性达到最大值;在nBPT各结构基团中,正丁基(-NH(CH2)3CH3)、硫基(-S)对nBPT与脲酶的结合起辅助作用,胺基(-NH2)是nBPT与脲酶结合的关键基团,与脲酶活性部位巯基(-SH)结合。土壤脲酶抑制剂nBPT与脲酶的具体结合机理还有待继续研究。     

氢醌对淹水土壤脲酶动力学特征的影响

通过模拟试验研究了正常水分和淹水条件下氢醌影响的土壤脲酶动力学特征。结果显示,培养时间、土壤类型、土壤水分及其相互作用显著影响土壤脲酶动力学参数的变化。与对照相比,HQ使土壤脲酶Km增加和Vmax降低,表明HQ属于典型的混合型抑制剂。随着培养时间延长,在正常水分和淹水条件下,HQ处理土壤脲酶Km降低,而Vmax和Vmax/Km升高。褐土脲酶Km的增加幅度及Vmax和Vmax/Km的降低幅度远远大于其它3种供试土壤。与正常水分相比,淹水对土壤脲酶动力学参数无显著性影响。相关分析表明,土壤脲酶动力学参数Km、Vmax和Vmax/Km与土壤理化性质存在显著的相关关系。     

乙草胺对土壤脲酶动力学特征的影响

采用室内模拟试验方法,以典型棕壤为供试土壤,研究不同浓度乙草胺对土壤脲酶活性和酶动力学参数的影响。结果表明,不同浓度乙草胺能显著抑制土壤脲酶活性,并在培养的第3～6 d达到最大抑制;利用模型 y=c/(1+bx)和y=c(1+ax)/(1+bx) 对不同浓度乙草胺与土壤酶活性的关系进行拟合,证明模型y=c/(1+bx)拟合效果较好,表明乙草胺除草剂对土壤脲酶的抑制作用为完全抑制,而脲酶ED50 为13.12～75.76 mg/kg;乙草胺的施入使土壤脲酶Vmax值降低,Km值则保持不变,属于典型的非竞争性抑制。     

陕西土壤脲酶与土壤肥力关系研究──Ⅱ.土壤脲酶的动力学特征

土壤脲酶动力学特征研究，国内少见报道。本文通过对陕西七种主要土壤十六个土样脲酶动力学参数的测定，揭示出：动力学参数（Ｖ０、Ｖｍ、Ｖｍａｘ、Ｖｍａｘ／Ｋｍ、Ｋ）和活化能Ｅａ从各个角度表征了处于不同生态环境中土壤脲酶的活性特性；高、低肥土样间也有明显差别，除水稻土外，各高肥土壤脲酶的Ｖ０、Ｖｍａｘ、Ｋ、Ｖｍａｘ／Ｋｍ均大于低肥，相关和主成分分析表明，动力学参数Ｖｍａｘ、Ｖｍａｘ／Ｋｍ可作为土壤肥力的指     

醋酸棉酚对土壤脲酶动力学特性的影响

通过不同浓度醋酸棉酚存在下的土壤脲酶催化尿素水解试验,研究醋酸棉酚对土壤脲酶的抑制作用,得到酶促反应的降解动力学参数和有关的热力学参数.结果表明,醋酸棉酚对土壤脲酶活性的抑制能力随醋酸棉酚浓度的增大而增强.由尿素浓度的倒数和水解速率的倒数为横纵标的双倒数曲线得到米氏常数(Km)和最大速率(Vmax),随着醋酸棉酚浓度的增大,酶促反应的Km和Vmax降低.在温度30℃时,Km由3.918 mol/L降到1.164 mol/L,Vmax由1.185×10-4 mol/(g·h)降到5.6×10-6 mol/(g·h).在醋酸棉酚的影响下,酶促反应的热力学参数包括活化能(Ea)、活化焓变(△H)和温度系数(Q10)有所下降.     

杀虫双对土壤脲酶活性特征的影响

通过模拟方法研究杀虫双对土壤脲酶活性特征参数的影响。结果表明 :不同生态区域土壤脲酶特征具有明显差异。杀虫双明显抑制脲酶活性 ,且随浓度增加 ,脲酶活性、酶促反应的Vmax、Vmax/Km、k减小 ,Km 增大 ,除 6号土样的Km 处理外均达到显著或极显著相关 ,揭示出脲酶特征参数可从不同角度表征杀虫双对土壤脲酶活性的影响 ,获得其作用机理为混合型抑制。脲酶活性ED50 值与土壤有机质、全氮和全磷呈现显著或极显著正相关关系 ,表明高有机质含量的土壤可明显减轻杀虫双的污染。     

脲酶抑制剂氢醌对土壤脲酶动力学行为的调控效应

研究了不同温度条件下脲酶抑制剂氢醌(HQ)对东北3种典型土壤(白浆土、棕壤、褐土)脲酶动力学参数的影响。结果表明,土壤类型、培养时间、培养温度及其相互作用均显著影响土壤脲酶动力学参数。与对照相比,加入HQ使土壤脲酶米氏常数(Km)增加,最大反应速率(Vmax)降低,表明HQ对土壤脲酶的作用机理属于混合型抑制。与白浆土相比,棕壤和褐土脲酶动力学参数受HQ的影响程度较大,表明高肥力土壤生物学活性较稳定。随着培养时间延长,土壤脲酶Km降低,Vmax和Vmax/Km增加。随着温度升高,土壤脲酶Km和Vmax增加,Vamx/Km无规律性变化。相关性分析表明,土壤脲酶动力学参数Km、Vmax和Vmax/Km与p H值、有机质、全氮、碱解氮和质地组成之间存在显著相关关系。     

不同土壤湿度下多菌灵的连续施用对土壤氮矿化速率与脲酶动力学的影响

明确在不同土壤水分环境下农药多菌灵对土壤氮循环和生物学属性的影响,对于评价多菌灵对土壤氮循环与生态环境的影响有重要意义。研究了在不同土壤湿度下连续添加不同浓度的多菌灵对土壤氮矿化、几丁质酶活性、土壤脲酶动力学参数以及微生物生物量的影响。结果表明:多菌灵添加量对土壤铵态氮、硝态氮、几丁质酶活性和微生物生物量氮具有显著影响;土壤湿度和多菌灵添加量的交互作用对铵态氮、硝态氮、矿化速率、脲酶酶促反应初速度(Vmax/Km)值以及微生物生物量氮有显著影响。在60%土壤最大持水量(WHC)条件下,5 mg·kg-1多菌灵的连续添加显著增加了铵态氮含量;在不同土壤湿度条件下,25 mg·kg-1多菌灵的连续添加显著增加了硝态氮含量。在60%WHC条件下连续添加10 mg·kg-1多菌灵的处理矿化速率显著高于不添加多菌灵的处理,但在90%WHC条件下添加10和25 mg·kg-1多菌灵处理的矿化速率显著低于不添加多菌灵的处理。在不同土壤湿度条件下,几丁质酶活性随多菌灵施用量增加而降低。在60%WHC条件下连续添加25 mg·kg-1多菌灵处理的脲酶Vmax/Km值显著低于连续添加10 mg·kg-1多菌灵的处理。多菌灵添加量可以通过影响土壤矿质氮含量与生物学属性从而影响土壤脲酶动力学的参数。     

全年淹水种植茭白对水田土壤性态的影响

水田全年淹水种植茭白是南方地区一种特色的土地利用方式,为了解其对土壤性状的影响,多点分别采集了全年淹水种植茭白(5 a以上)与长期种植水稻的水田耕层土壤样品,比较了它们之间在土壤理化性状上的差异;同时以长期种植水稻的水田为对照,选择种植茭白4、12、19 a的3块同类型土壤的水田,观察分析了土壤剖面的物理、化学和形态学性状,探讨水田长期种植茭白后土壤性态的演变趋势。结果表明,水田全年淹水种植茭白后耕层土壤有机质、全氮、全磷、有效磷和速效钾均呈现明显的增加,但土壤结构逐渐退化,土壤分散性增强,并容易出现黏闭、起浆、僵硬和水气矛盾等问题。全年淹水种植茭白后,水田土壤犁底层逐渐变薄最终消失,上部软糊土层加厚,种植茭白19 a时软糊土层厚度已达40 cm;全年淹水种植茭白20～40 cm土层有机质含量明显地增加,0～40 cm土层土壤氧化还原电位下降,土色变暗,无定形氧化铁和铁的活化度显著地增加;相应地土壤类型也逐渐由水耕人为土向滞水潜育土演变。     

Cr^3＋对土壤脲酶活性特征的影响

采用模拟方法对Cr^3＋的土壤脲酶效应进行了研究,结果表明,土壤pH对Cr^3＋的生态毒性有重要影响;酸性土壤脲酶受到显著抑制,活性及动力学特征参数与Cr^3＋浓度间达显著或极显著负相关,而且模型U=β0（/β1×C＋1）揭示其间机理为完全抑制,动力学则进一步细化为非竞争性抑制;获得土壤轻微和中度污染时的生态剂量ED10和ED50分别为50.59和865.7 mg.kg^-1;酸性土壤中脲酶活性、Vmax、k可作为土壤Cr^3＋污染的监测指标之一,而碱性土壤则反应不敏感,其随铬浓度增加,脲酶活性及动力学参数呈先增加后降低的规律性变化,总体变幅较小;两类土壤的差别可能主要是由于土壤环境引起了不同价态铬转变的缘故。     

Stability of stored N-(n-butyl) thiophosphoric triamide (NBPT) treated urea-based fertilizers

ABSTRACT

The effectiveness of N-(n-butyl) thiophosphoric triamide (NBPT) in reducing ammonia volatilization from urea-based fertilizers has been thoroughly investigated. However, the stability of this inhibitor during storage of NBPT treated urea and urea ammonium nitrate (UAN) needs further investigation. We compared ammonia volatilization from NBPT treated urea (360 mg NBPT kg^?1 urea) and UAN (180 mg NBPT L^?1 UAN) that were stored at room temperature for 6, 3 and 0 months. We measured ammonia volatilization with cylindrical chambers fitted with acid-charged discs at five times for 21 d. Total ammonia volatilization (measured as a % of applied nitrogen) was significantly greater in untreated urea and UAN (32% to 33%) than those in NBPT treated urea and UAN (6% to 12%). Reduction of ammonia volatilization was not significantly different among NBPT treated urea (73% to 81%) and UAN (63% to 73%) irrespective of storage time. This implies that farmers can mix their urea-based fertilizers with NBPT formulation 6 months prior to fertilization without compromising the ammonia volatilization reducing property of the NBPT.     

Effect of N-(n-butyl) thiophosphoric triamide on hydrolysis of urea by plant,microbial, and soil urease

Summary Comparison of the effects of N-(n-butyl) thiophosphoric triamide (NBPT) and phenylphosphorodiamidate (PPD) on hydrolysis of urea by plant (jackbean), microbial (Bacillus pasteurii), and soil urease showed that whereas NBPT was considerably more effective than PPD for inhibiting hydrolysis of urea added to soil, it was much less effective than PPD for inhibiting hydrolysis of urea by plant or microbial urease. Studies to account for this observation indicated that NBPT is rapidly decomposed in soil to a compound that is much more effective than NBPT for inhibition of urease activity and that this compound is N-(n-butyl) phosphoric triamide.     

土壤温度和含水量互作对抑制剂抑制氮素转化效果的影响

为比较生化抑制剂组合对土壤氮素转化的抑制效果,揭示不同土壤温度和含水量互作对尿素水解抑制效应的影响。该文采用室内模拟培养方法,研究土壤含水量(60%和80%田间最大持水量,water holding capacity,WHC)和土壤温度(15、25和35℃)互作对生化抑制组合[N-丁基硫代磷酰三胺(N-(n-butyl)thiophosphoric triamide,NBPT)、N-丙基硫代磷酰三胺(N-(n-propyl)thiophosphoric triamide,NPPT)和2-氯-6(三氯甲基)吡啶(2-chloro-6(trichloromethyl)pyridine,CP)在黄泥田土壤中抑制氮素转化效果的影响。结果表明:土壤温度和含水量对生化抑制组合在黄泥田土壤中抑制尿素水解效应显著,以土壤温度影响更大。随着土壤温度增加,尿素水解转化增强,有效作用时间降低,硝化作用增强,脲酶和硝化抑制效应减弱;随着土壤含水量降低,尿素水解转化缓慢,有效作用时间延长,硝化作用减弱,脲酶和硝化抑制效应增强。不同土壤温度和含水量条件下,NBPT/NPPT或配施CP处理有效抑制黄泥田土壤脲酶活性,延缓尿素水解;CP或配施NBPT/NPPT处理有效抑制NH4+-N向NO_3~--N转化,保持土壤中较高NH_4~+-N含量长时间存在。新型脲酶抑制剂NPPT单独施用及与CP配施的土壤尿素水解抑制效果与NBPT相似。黄泥田土壤中生化抑制组合应用最佳的土壤温度和含水量分别为25℃和60%WHC。总之,针对不同土壤温度和含水量条件,在黄泥田土壤中应采用脲酶抑制剂与硝化抑制剂相结合的施肥方式。     

Rate and mode of application of the urease inhibitor N-(n-butyl) thiophosphoric triamide on ammonia volatilization from surface-applied urea

A laboratory study evaluated the effect of rate (0, 100, 250, 500, 750 or 1000 mg/kg) and mode of application of the urease inhibitor N -( n -butyl) thiophosphoric triamide (nBTPT) (coating the urea granule, adding to the urea melt or adding to urea ammonium nitrate (UAN) solutions) on NH₃ volatilization from urea, at three temperatures (5, 15 and 25 °C), with four contrasting soil types. Daily ammonia loss was measured for up to 21 days after surface N application, using ventilated soil enclosures. Ammonia loss from unamended urea varied with soil type and temperature and ranged from 8.2 to 31.9% of the N applied. nBTPT was highly effective in lowering NH₃ volatilization from urea and in delaying the time of maximum rate of loss. The average % inhibition over all soils, temperatures and formulations was 61.2, 69.9, 74.2, 79.2 and 79.8% for the 100, 250, 500, 750 or 1000 mg/kg nBTPT concentration, respectively. The % inhibition with nBTPT was lower at 15 °C compared with at 5 or 25 °C and was lower in UAN solutions than in granular products. There was little difference between the melted and coated granular products in lowering NH₃ loss or in soil N transformations. The stability of nBTPT in urea products was dependent on its mode of application and on the storage temperature. Incorporating nBTPT in the urea melt produced a more homogeneous product with superior stability than coating the urea granule.     

Nitrogen Fertilization of Soybean Affects Root Growth and Nodulation on Two Soil Types in Mississippi

Greenhouse studies were conducted to evaluate the influence of nitrogen (N) sources [urea + ?N-(n-butyl) thiophosphoric triamide, NBPT (urease inhibitor) and polymer-coated urea (PCU)] and rates on soybean root characteristics, nodule formation, and biomass production on two soil types (silt loam and clay) commonly cropped to soybean in Mississippi. About 15% less belowground biomass was produced in clay soil than in silt loam soil directly corresponding to all other root parameters including root length, root area, root diameter, and nodule number. Pooled across N rates, N additions resulted in 19% and 52% decrease in belowground biomass and number of nodules, respectively, across soils compared to soybean receiving no N. The N rate was the most critical factor as it influenced all root growth parameters. Number of nodules were 24% greater with PCU than urea + NBPT. Nitrogen additions and clay soil negatively impacted soybean root growth, nodulation, and belowground biomass production.

Abbreviations: Polymer-coated urea, PCU; N-(n-butyl) thiophosphoric triamide, NBPT     

Enhanced-efficiency nitrogen fertilizers reduce winter losses of nitrous oxide,but not of ammonia,from no-till soil in a subtropical agroecosystem

Nitrogen (N) gas losses can be reduced by using enhanced-efficiency N (EEN) fertilizers such as urease inhibitors and coating technologies. In this work, we assessed the potential of EEN fertilizers to reduce winter losses of nitrous oxide (N₂O-N) and ammonia (NH₃-N) from a subtropical field experiment on a clayey Inceptisol under no-till in Southern Brazil. The EEN sources used included urea containing N-(n-butyl) thiophosphoric triamide (UR+NBPT), polymer-coated urea (P-CU) and copper-and-boron-coated urea (CuB-CU) in addition to common urea (UR) and a control treatment without N fertilizer application. N₂O-N and NH₃-N losses were assessed by using the static chamber method and semi-open static collectors, respectively. Both N₂O-N and NH₃-N exhibited two large peaks with an intervening period of low soil moisture and air temperature. Although the short-term effect was limited to the first few days after application, UR + NBPT urea decreased soil N₂O-N emissions by 38% relative to UR. In contrast, urease inhibitor technology had no effect on NH₃-N volatilization. Both coating technologies (CuB-CU and P-CU) were ineffective in reducing N losses via N₂O production or NH₃ volatilization. The N₂O emission factor (% N applied released as N₂O) was unaffected by all N sources and amounted to only 0.48% of N applied—roughly one-half the default factor of IPCC Tier 1 (1%). Based on our findings, using NBPT-treated urea in the cold winter season in subtropical agroecosystems provides environmental benefits in the form of reduced soil N₂O emissions; however, fertilizer coating technologies provide no agronomic (NH₃) or environmental (N₂O) advantages.     

Urea persistence in floodwater and soil used for flooded rice production

Urea is a common fertilizer in delayed‐flood rice production in the United States, and its use worldwide has increased dramatically in recent decades. This study aimed to directly quantify urea‐N persistence in floodwater and soil used for rice production. We conducted a set of three laboratory experiments to investigate urea‐N presence in the floodwater and soil. Untreated urea was applied to dry or wet soil and flooded immediately or urea treated with the urease inhibitor N‐(n‐butyl)‐thiophosphoric triamide (NBPT), or untreated urea was applied to dry soil and flooded after a 5‐day delay. Urea‐N was analysed colorimetrically (using the microplate‐adapted, diacetyl monoxime method) in the floodwater, and at 2‐cm intervals in soil after 10‐cm long, silt‐loam soil columns were flooded for 12, 24, 48 and 96 h. The only management practice that led to insignificant urea‐N concentrations in floodwaters was the application of urea followed by a 5‐day delay before flooding. Urea‐N can persist in floodwaters for an estimated 98 and 120 h after immediately flooding dry‐soil‐applied or wet‐soil‐applied untreated urea, respectively. Urea‐N concentrations in floodwaters were up to 33 times less when dry‐soil‐applied than wet‐soil‐applied. Average NBPT‐treated urea‐N concentrations in soil ranged up to 63 mg/kg after 24 h of flooding and were <1 mg/kg after 96 h of flooding. The 5‐day delay resulted in ≤1 mg urea‐N/kg soil when untreated urea was applied. Generally, the threat of N entering adjacent waterways in the form of urea is likely to be limited because of its short‐term persistence (≤120 h) in rice floodwater.     

Effects of the urease inhibitor N-(n-butyl)phosphorothioic triamide in low concentrations on ammonia volatilization and evolution of mineral nitrogen

Laboratory incubation experiments were conducted to study the influence of increasing concentrations of N-(n-butyl)phosphorothioic triamide (NBPT) on NH₃ volatilization and rate of urea hydrolysis and evolution of mineral N in Ozzano, Rimini and Carpi soils with different physicochemical characteristics. Low concentrations of NBPT reduced NH₃ losses due to volatilization after urea fertilization and the effectiveness of the inhibitor was related to the soil characteristics (e.g. high concentrations of organic C and sand). After 15 days of incubation, no significant reductions of losses were found for any of the NBPT concentrations employed in Rimini soil. The application of NBPT led to a considerable reduction of the formation of nitrite. This process was completely annulled with the highest dose of NBPT (0.5% w/w_urea) in the Carpi soil after 15 days. In Rimini soil, however, the use of NBPT was less effective in influencing nitrite formation. The use of NBPT favoured accumulation of nitrate proportional to the NBPT concentration employed while it had no influence on the NH _inf4 ^sup+ fixation by 2:1 layer silicates. The data obtained support previous evidence that NBPT is effective in reducing the problems encountered in using urea as fertilizer. However, environmental conditions and soil physicochemical characteristics may have an important influence on the effectiveness of NBPT.