脲酶抑制剂氢醌对大豆结瘤、固氮和木质部溶质氮形态的影响

首次报导了在白浆土和砂壤土上使用氢醌(HQ)抑制土壤脲酶活性,有效地缓解尿素分解产物及其随后的氧化产物对大豆结瘤和固氮活性的抑制效应,结果表明:1.HQ浓度在40PPM以内对大豆幼苗生长及初生结瘤表现促进作用;进一步提高HQ浓度将使大豆根系生长受阻变态而阻止结瘤。2.HQ(10—50PPM)提高了离体活性大豆根瘤类苗体悬液的耗氧量(79.4—86.1％)和琥珀酸脱氢酶活性(124.7—138.4％)。3.盆栽和田间试验证实,由于HQ缓解了尿素的分解,从而颇大减轻了尿素对大豆结瘤和固氮(乙炔还原活性)的抑制效应;通过大豆木质部中溶质氮形态(酰胺、酰脲和硝酸盐)的分析进一步证实了,大豆植株从根部向地上部运输的氮素形态同土壤氮转化强度和根瘤固氮强度(酰脲相对丰度)之间的紧密联系。4.由于麦秸还田土壤脲酶活性提高,故应提高HQ剂量;与此同时,通过麦秸的“氮因子效应”便能完全解除尿素对大豆结瘤固氮的抑制,并为大豆籽实发育提供了丰富的土壤氮源。     

柑桔园土壤脲酶活性年周期变化及脲酶抑制剂氢醌的效应

本研究初步探明南亚热带赤红壤柑桔园土壤脲酶活性的年周期变化:冬季最低,春季升至高峰,夏季开始下降,直至秋季仍处于较低水平。室内模拟试验表明:氢醌对柑桔园土壤脲酶活性有明显抑制作用,其抑制率随浓度提高而增加,且随着培养时间的延长而逐渐下降;添加氢醌处理的尿素水解持续时间,比不加氢醌的延长约1倍。     

脲酶抑制剂氢醌的环境效应评价

本文根据用标记和非标记氢醌进行的模拟、盆栽和田间定位试验,结合国内外文献有关氢醌的环境常数,论述了氢醌在土壤-植物系统的去向和代谢途径、对土壤酶活性的影响及其环境效应。得出的结论是:作为脲酶抑制剂使用的微量氢醌(0.3—0.4％,与尿素重量比),不会从土壤中淋失和挥发,在土壤和植物中没有累积,对与碳、氮和磷转化有关的土壤酶活性很少影响。在土壤中,它将通过氧化、臭氧化和生物学降解,经由环断裂生成二元酸或参与腐殖物质的合成。在植物体内,主要通过糖苷化得到同化和利用。因此,氢醌作为脲酶抑制剂在农业生产中应用是安全的。     

秸秆及秸秆黑炭对小麦养分吸收及棕壤酶活性的影响

通过小麦盆栽试验,研究了玉米秸秆及其秸秆黑炭施加对小麦养分吸收利用和棕壤酶活性的影响。试验设对照(CK),黑炭(B),秸秆还田(S),尿素(U),尿素+黑炭(UB)及尿素+秸秆还田(US)6个处理,各处理3次重复。结果表明:无氮肥施入下,B处理较CK和S处理籽粒产量显著提高99.4%和77.7%,小麦地上部氮、磷、钾吸收累积量分别显著提高94.1%—140.9%,55.4%—66.3%和53.1%—72.6%;有氮肥施入下,UB和US处理较U处理提高籽粒产量8.2%—8.8%,小麦地上部氮、磷、钾吸收累积量分别显著提高14.3%—27.8%,19.6%—30.9%和24.4%—40.9%。秸秆及秸秆黑炭施加处理的氮素利用率显著提高21.4%—41.7%。黑炭施加显著提高土壤中有机碳、NH+4-N、NO-3-N和速效钾含量;在施氮条件下,秸秆还田显著提高土壤中NO-3-N含量;秸秆及黑炭施加对有效磷含量无显著影响。秸秆还田显著提高了土壤脱氢酶、过氧化氢酶、脲酶和中性磷酸酶活性;施加黑炭也明显提高了土壤脱氢酶和脲酶活性,但抑制过氧化氢酶和中性磷酸酶活性。土壤脲酶活性与土壤有机碳、无机氮含量呈显著正相关,表明土壤酶可反映土壤肥力水平。     

汞、镉对土壤脲酶活性影响的研究Ⅰ.尿素浓度

对不同尿素浓度条件下重金属与土壤脲酶活性关系进行了研究。结果表明,尿素浓度对脲酶活性具有显著的影响。在供试浓度范围内可采用线性和Langmuir模型较好地表征二者关系。并得到脲酶活性的尿浓贡献率,尿浓贡献变化率和最大表观脲酶活性等参数;Hg,Cd明显降低了前述参数值,其中Hg Cd复合污染的抑制作用最强,Hg的生态毒性最大;同时初步获得土壤酶促反应过程中存在吸附机制。     

脲酶抑制剂和硝化抑制剂的协同作用对尿素氮转化和N2O排放的影响

根据培养试验,论述了脲酶抑制剂氢醌和硝化抑制剂双氰胺和碳化钙的不同组合在土壤正常水分和渍水的条件下对于土中尿素的水解及其释出的氨的吸附、氧化和挥发以及Ｎ２Ｏ生成的影响．文章指出,配合使用氢醌和双氰胺既能延缓土中尿素的水解并使水解后释出的氨在土中得以更多和更长时间的保持,还能减少土中硝酸盐的累积、氨挥发的损失及Ｎ２Ｏ的生成．这表明在脲酶抑制剂和硝化抑制剂间可能存在一定的协同作用．很好利用这一作用,将有益于提高尿素肥效和减少其Ｎ损失与环境污染．     

碳添加下黑钙土胞内、胞外脲酶活性变化及其机制

土壤脲酶作为能够催化尿素水解的最重要酶类,对草地生态系统氮素供应具有重要作用。目前探讨不同碳添加对草地土壤胞外脲酶影响的研究报道相对较多,但碳添加对土壤胞内脲酶的影响,以及胞内和胞外脲酶对碳添加的响应是否一致等尚需深入研究。本研究依托额尔古纳森林草原过渡带生态系统研究站开展的碳添加野外试验平台(以葡萄糖为碳源),选取无碳添加(C₀)、250(C₂₅₀)和500(C₅₀₀) kg C·hm^-2·a^-1处理为供试对象,探讨碳添加下黑钙土胞内、胞外脲酶活性响应及其与土壤性质的关系。结果表明: 碳添加显著提高了土壤胞内脲酶活性,增加了土壤胞内脲酶活性占总脲酶活性的比例,但对土壤胞外脲酶活性没有显著影响。土壤胞内脲酶活性与微生物生物量具有显著正相关关系,表明胞内脲酶活性增加主要是由微生物生物量增加引起的。结构方程模型(SEM)分析表明,碳添加通过影响土壤微生物生物量间接提高了土壤胞内脲酶活性。     

脲酶-硝化抑制剂对减缓尿素转化产物氧化及淋溶的作用

利用原状土柱模拟试验,研究了脲酶抑制剂氢醌(HQ),硝化抑制剂包被碳化钙(ECC)和双氰胺(DCD)以及它们的不同组合对尿素转化产物土壤持留、氧化以及淋溶的影响.结果表明,与其它抑制剂处理相比,HQ+DCD组合能有效抑制尿素水解产物的氧化,使其以交换态NH₄⁺的形式在土壤中长时间持留;氧化作用的抑制不仅减少了氧化产物NO₃^-的累积,也降低了NO₃^-淋溶潜势,使其淋入下层土壤的深度仅限在5～10 cm范围内,且淋溶量显著降低.     

脲酶-硝化抑制剂对减缓尿素转化产物氧化及淋溶的作用

利用原状土柱模拟试验,研究了脲酶抑制剂氢醌(HQ),硝化抑制剂包被碳化钙(ECC)和双氰胺(DCD)以及它们的不同组合对尿素转化产物土壤持留、氧化以及淋溶的影响.结果表明,与其它抑制剂处理相比,HQ+DCD组合能有效抑制尿素水解产物的氧化,使其以交换态NH₄⁺的形式在土壤中长时间持留;氧化作用的抑制不仅减少了氧化产物NO₃^-的累积,也降低了NO₃^-淋溶潜势,使其淋入下层土壤的深度仅限在5～10 cm范围内,且淋溶量显著降低.     

春季海南岛近岸海域尿素与浮游生物的脲酶活性

2011年春季(4-5月),对海南岛的海口湾、澄迈湾、文昌八门湾、陵水新村湾和大东海5个海湾的尿素浓度及浮游生物的脲酶活性开展调查研究,结合其它理化环境因子,分析海南岛近岸海域尿素的可利用性及其对该海区浮游植物生长可能产生的影响.结果表明,海南岛近岸水体中尿素平均浓度为2.07-3.30 μmol/L,占总溶解态氮TDN含量的14％-38％,尿素占TDN比例由北向东、南方向递增.浮游生物脲酶活性为0.30-0.84 μmolN· L-1· h-1,海口湾最高,从北部向东、南部逐渐减少.各海湾较高水平的尿素和脲酶活性主要分布在排污口、养殖区或旅游区的近岸海域.硅藻为优势种,甲藻种类少且密度低,部分甲藻密度达到104-105个/L的水体,尿素和脲酶活性也处于较高水平.海区浮游植物细胞密度与脲酶活性或尿素占TDN比例等因子存在相关性,表明尿素是海南海域浮游植物生长不可忽略的重要氮源.尿素在一定程度上促进春季海南岛近岸海域甲藻等浮游植物的生长,可能对浮游植物群落结构的改变产生重要影响.     

氢醌和双氰胺对种稻土壤N2O和CH4排放的影响

通过盆栽试验,研究了脲酶抑制剂氢醌（ＨＱ）、硝化抑制剂双氰胺（ＤＣＤ）及二者的组合（ＨＱ＋ＤＣＤ）对种稻土壤Ｎ２Ｏ和ＣＨ４排放的影响．结果表明,在未施麦秸粉时,所有施抑制剂的处理均较单施尿素的能显著减少水稻生长期供试土壤Ｎ２Ｏ和ＣＨ４的排放．特别是ＨＱ＋ＤＣＤ处理,其Ｎ２Ｏ和ＣＨ４排放总量分别约为对照的１／３和１／２．而在施麦秸粉后,该处理的Ｎ２Ｏ排放总量为对照的１／２,但ＣＨ４排放总量却较少差别．不论是Ｎ２Ｏ还是ＣＨ４的排放总量,施麦秸粉的都比未施的高出１倍和更多．因此,单从土壤源温室气体排放的角度看,将未腐熟的有机物料与尿素共施,并不是一种适宜的施肥制度．供试土壤的Ｎ２Ｏ排放通量,与水稻植株的ＮＯ－３Ｎ含量和土表水层中的矿质Ｎ量分别呈显著的指数正相关和线性正相关;ＣＨ４的排放通量则与水稻植株的生长量和土表水层中的矿质Ｎ量呈显著的线性负相关．在Ｎ２Ｏ与ＣＨ４的排放间,未施麦秸粉时存在着定量的相互消长关系;施麦秸粉后,虽同样存在所述关系,但难以定量化．     

Leaf urea metabolism in potato. Urease activity profile and patterns of recovery and distribution of (15)N after foliar urea application in wild-type and urease-antisense transgenics

The influence of urease activity on N distribution and losses after foliar urea application was investigated using wild-type and transgenic potato (Solanum tuberosum cv Désirée) plants in which urease activity was down-regulated. A good correlation between urease activity and (15)N urea metabolism (NH(3) accumulation) was found. The general accumulation of ammonium in leaves treated with urea indicated that urease activity is not rate limiting, at least initially, for the assimilation of urea N by the plant. It is surprising that there was no effect of urease activity on either N losses or (15)N distribution in the plants after foliar urea application. Experiments with wild-type plants in the field using foliar-applied (15)N urea demonstrated an initial rapid export of N from urea-treated leaves to the tubers within 48 h, followed by a more gradual redistribution during the subsequent days. Only 10% to 18% of urea N applied was lost (presumably because of NH(3) volatilization) in contrast to far greater losses reported in several other studies. The pattern of urease activity in the canopy was investigated during plant development. The activity per unit protein increased up to 10-fold with leaf and plant age, suggesting a correlation with increased N recycling in senescing tissues. Whereas several reports have claimed that plant urease is inducible by urea, no evidence for urease induction could be found in potato.     

Urease activity inTrichophyton mentagrophytes

Urease activity was detected in the dermatophyteTrichophyton mentagrophytes cells at early exponential phase of growth. Specific activity of urease decreased with culture age. At exogenous urea concentrations above 2 mm formation of urease was inhibited. The pH optimum lay at 7–7.5, the K_m being 14 mm. No urease activity could be detected in cell-free culture fluid ofT. mentagrophytes. No endoor exocellular urease activity could be detected in aT. rubrum strain grown with or without urea.     

Biological changes in the rhizosphere of wheat after foliar application of chlorocholinechloride,urea and 4-chloro-2-methylphenoxyacetic acid

In field experiments wheat in the phase of shooting was sprayed with solutions of chlorocholinechloride (CCC) and urea, CCC and ammonium salt MCPA (Aminex) or CCC, urea and Aminex. The effect of the treatment on dry weight of overground parts of wheat, number of bacteria, production of carbon dioxide, urease activity and content of ammonium in the rhizosphere soil was investigated. In all cases evolution of carbon dioxide in the rhizosphere soil was higher than that in the control soil. Highest numbers of bacteria were found in the rhizosphere soil of plants treated with urea, the herbicide and their mixtures. Content of ammonium was higher in the control soil than in the rhizosphere soils, the urease activity was highest in the rhizosphere soil of plants treated with the solution of the herbicide and with the combination of the herbicide with urea.     

与氮转化有关的土壤酶活性对抑制剂施用的响应

利用室内模拟培养试验,研究好气条件下施用尿素后土壤脲酶、脲酸还原酶、亚硝酸还原酶和羟胺还原酶活性对脲酶抑制剂氢醌（HQ）与硝化抑制剂包被碳化钙（ECC）和双氰胺（DCD）组合（HQ ECC、HQ DCD）的响应、结果表明,HQ DCD组合与其它抑制剂处理相比能更有效地降低土壤脲酶活性,增加硝酸还原酶、亚硝酸还原酶、羟胺还原酶活性,不同处理土壤脲酶、亚硝酸还原酶和羟胺还原酶活性与土壤NH4^ 、NO3^-、NH3挥发和N2O排放速率间存在不同形式的显著相关关系：土壤脲酶、亚硝酸还原酶和羟胺还原酶活性之间存在不同形式的显著正相关关系。     

Urease Activity in Aspergillus tamarii

Previous (unpublished) studies of the vegetative and conidialdevelopment of several fungi of the genus Aspergillus grownon different nitrogenous substrates showed that urea servedas a good source of nitrogen in all cases investigated. Moredetailed work carried out with Aspergillus tamarii IMI 61268has failed to establish the presence of extracellular or cellbound urease activity but shows high levels of activity in aqueousextracts of this fungus. In surface-grown, sporulating cultures,urease activity increased prior to conidiation and disappearedrapidly during the reproductive phase. Similar patterns of enzymicactivity were obtained with cultures grown on urea and ammoniaas sole sources of nitrogen. In submerged, shaken flask cultures using ammonia as the solenitrogenous substrate, conidiation was inhibited and ureaseactivity continued to increase until such time as the residualammonia in the medium became depleted. Thereafter the ureaseactivity as well as total soluble protein showed a sharp decrease.The results suggest that the enzyme showing urease activityin this organism is not substrate-inducible and one of its functionsmay be that of a storage protein whose breakdown products assistin the continued growth and especially in conidial development.     

Urease testing and yeast taxonomy

When urease production was assayed by the hydrolysis of [14C]urea, all basidiomycetous yeasts tested, including the Cryptococcus vishniacii complex (previously reported urease negative), produced significant amounts of 14CO2. The Schizosaccharomycetaceae were the only urease-positive ascomycetous yeasts tested. Yarrowia lipolytica was urease negative. The stoichiometry of [14C]urea hydrolysis paralleled by Roberts' rapid urea hydrolysis (RUH) test indicated that causes of anomalous results in conventional urease testing include acidification and alkalinization of the test medium by products of endogenous metabolism and autolysis rather than urease activity. Anomalous results also occurred when cells were grown on media containing the chelating agent ethylenediaminetetraacetic acid (EDTA) prior to RUH. The addition of EDTA to a complex natural medium inhibited urease production in all yeasts reportedly growing at 35 degrees C (and all other yeasts tested), except Filobasidiella (Cr.) neoformans var. neoformans (NIH 12). The RUH test could differentiate at the varietal level: Fil. (Cr.) neoformans var. neoformans was about 10 times more resistant to EDTA in media used for the growth of cells prior to RUH testing than was Fil. neoformans var. bacillispora (Cr. neoformans var. gattii) (NIH 191). Urease production by Fil. neoformans var. bacillispora was specifically restored to half maximal activity by the addition of 22 microM Ni+2 (as NiCl2) to a growth medium containing 0.100 mM EDTA.     

Molecular Modeling and docking of Wheat Hydroquinone Glucosyl transferase by using Hydroquinone,Phenyl phosphorodiamate and n-(n butyl) Phosphorothiocic Triamide as Inhibitors

In agriculture high urease activity during urea fertilization causes substantial environmental and economical problems by releasing abnormally large amount of ammonia into the atmosphere which leads to plant damage as well as ammonia toxicity. All over the world, urea is the most widely applied nitrogen fertilizer. Due to the action of enzyme urease; urea nitrogen is lost as volatile ammonia. For efficient use of nitrogen fertilizer, urease inhibitor along with the urea fertilizer is one of the best promising strategies. Urease inhibitors also provide an insight in understanding the mechanism of enzyme catalyzed reaction, the role of various amino acids in catalytic activity present at the active site of enzyme and the importance of nickel to this metallo enzyme. By keeping it in view, the present study was designed to dock three urease inhibitors namely Hydroquinone (HQ), Phenyl Phosphorodiamate (PPD) and N-(n-butyl) Phosphorothiocic triamide (NBPT) against Hydroquinone glucosyltransferase using molecular docking approach. The 3D structure of Hydroquinone glucosyltransferase was predicted using homology modeling approach and quality of the structure was assured using Ramachandran plot. This study revealed important interactions among the urease inhibitors and Hydroquinone glucosyltransferase. Thus, it can be inferred that these inhibitors may serve as future anti toxic constituent against plant toxins.     

Fate of urea-15N in a soil-wheat system as influenced by urease inhibitor hydroquinone and nitrification inhibitor dicyandiamide

By applying labeled urea into a loamy meadow brown soil, a pot experiment with spring wheat as test crop was carried out. The results showed that at the end of this experiment, the plant recovery, the soil recovery and the total loss of applied urea ¹⁵N was 17.7–23.7%, 43.7–56.3% and 20.0–36.8%, respectively. ¹⁵N recovery by wheat grain in any treatment varied within a range of 9.0–14.7% of the applied ¹⁵N. A combined application of hydroquinone (HQ) and dicyandiamide (DCD) gave the lowest loss and the highest recoveries in both the plant and soil, while applying HQ or DCD alone had less effect on them. During the whole period of wheat growth, HQ+DCD induced an increasing ¹⁵N uptake by plant, and even promoted the translocation of absorbed ¹⁵N from stem to grain. In the presence of inhibitors, organic plus chemically fixed ¹⁵N occupied a large portion of soil ¹⁵N recovery at maturity stage of wheat growth (34.3–50.6%, in contrast to 9.9% in the absence of inhibitors), and DCD and DCD+HQ could remarkably reduce the remaining soil (NO₃ ^-+NO₂ ^-)-¹⁵N. In this pot experiment, the leaching loss of applied ¹⁵N was excluded, and hence, the gaseous loss was considered as the main part of the ¹⁵N loss. Regarding N loss, N₂O flux only occupied a very small part, and its main part was other gaseous N losses. DCD and DCD+HQ retarded N₂O flux from the soil-wheat system after treatment with urea and reduced the total N₂O flux during the whole period of wheat growth. Treatment with both inhibitors had much lower gaseous N losses than that with HQ or DCD alone. Hence, a proper combination application of HQ and DCD is an efficient way to improve urea-N efficiency and crop quality, while decreasing its loss to the environment. This revised version was published online in June 2006 with corrections to the Cover Date.