小麦叶内硝酸还原的代谢库

随营养液中No_3~-浓度升高,叶片内No_3~-总量、代谢库大小(NIPS)及硝酸还原酶(NR)活性均升高,其中MPS与NK活性呈同步变化;No_3~-浓度达2.0mmol/L时,两者趋于稳值;若再增加NO_3~-浓度,则被吸收的NO_3~-积累于液泡中,而代谢库中NO_3~-含量(MPS)与NO_3~-总量之比有一定程度降低。低氮(NO_3~-浓度为1.0 mmol/L)情况下,反应液中无NO_3~-时,叶片内NR活性品种间有差异,但在50 mmol/L NO_3~-反应液中则品种间无差异;NK活性高的品种鲁麦8号及品种321叶内有大的NO_3~-代谢库,反应液中NO_3~-对NR活性刺激程度低,代谢库NO_3~-含量与叶NO_3~-总量之比高,而叶组织长时间反应过程中其NR活性衰减速率低。     

玉米幼苗NO_3~-的吸收、溢泌和硝酸还原酶活性在品种间的差异

玉米品种间NO_3~-吸收的表观米氏常数(K_m,app)、最大吸收速率(I_m)有明显的差异。品种813NO_3~-吸收速率大于中单2号;溢泌液体积及其NO_3~-含量也是这样。硝酸还原酶(NR)的体外测定表明,地上部的活性比根部的大得多;不论地上部或根部的NR活性(NRA),品种813的大于中单2号。NRA的体内测定表明,去胚乳和盾片的幼苗经诱导,反应液有NO_3~-,813的第1叶的NRA大于中单2号;不去胚乳和盾片幼苗的第1叶NRA中单2号大于813。     

植物体内NO3^—可给性对硝酸还原酶活性的调节

评述植物叶片中NO_3~-可给性对活体硝酸还原酶(NR)活性的调节,指出根部NO_3~-的不断供给及液泡内NO_3~-的外流可以使细胞质内的NO_3~-维持一定水平,这对NR的诱导及整个NO_3~-还原系统高活力的稳定是必需的。NO_3~-对NR的诱导反映在NR的mRNA转录水平上。     

NO_3~-转运蛋白在植物适应逆境中的功能研究进展

氮是植物生长发育所必需的大量元素,参与植物体内各种代谢活动。硝态氮(NO_3~-)是植物可吸收利用的主要无机氮源。NO_3~-转运蛋白不仅介导植物正常生长发育过程中NO_3~-的吸收、转运和再利用,还参与调控植物对多种逆境的适应过程。结合最新报道,重点总结了近年来关于NO_3~-转运蛋白在植物适应低NO_3~-、低K+、盐、干旱及重金属镉等胁迫中的重要作用的研究进展,以期为今后进一步深入探究植物抗逆机理提供依据和参考。     

稀土元素对小麦幼苗氮素吸收及其同化的影响

稀土元素浸种能够促进小麦(Triticum aestivum L.)幼苗对NO_3~-的吸收,提高硝酸还原酶活力。这些效应与稀土浓度有关,低浓度有促进作用,高浓度则有抑制作用。稀土元素处理还能促进小麦幼苗体内NO_3~-的同化还原,使硝态氮含量降低,氨基氮含量增加,促进了氮素代谢过程。     

春化对油菜叶片硝酸还原酶活性的影响

油莱种子经春化后,其子叶NR活性提高了1.5～4.9倍,真叶展开、雌雄分化及开花期的NR活性也高于对照。去顶芽、高温中断及抑制剂处理证实春化是导致NR活性变化的原因。春化期间幼苗根部对NO_3~-吸收及叶片ATP含量的增加可能是NR活性升高的部分生理机制。     

超氧物歧化酶活性与小麦对HSO3^—,NO2^—抗性之间的关系

用在培养液中加低浓度NO_2~-,HSO_3~-,Mn~(2 )及向地上部喷洒氯霉素等四种方法促使小麦苗超氧物歧化酶(SOD)活性增加。SOD活性提高后,植株叶片对高浓度NO_2~-,HSO_3~-的抗性增强,表现在因NO_2~-,HSO_3~-害而产生的膜脂过氧化产物乙烷减少,叶绿素分解减轻。低浓度NO_2~-处理后的麦苗,不仅提高了对高浓度NO_2~-的抗性,同时对HSO_3~-的抗性也增强;同样,低浓度HSO_3~-处理也同时促进了麦苗对HSO_3~-和NO_2~-的抗性,即可交叉地提高。这些结果直接说明SOD和抗性有关。同时也为SO_2/HSO_3~-,NO_2/NO_2~-害的自由基学说提供了一个佐证。     

NaCl胁迫下硝酸盐对大麦幼苗根系Cl~-吸收的调节(简报)

在含20mmolL~(-1) 的 NaCl溶液中,当加入的NO_3~-浓度由0增加到10mmol L~(-1)时,两个耐盐性不同的大麦品种幼苗根系Cl~-吸收速率呈明显降低趋势。NO_3~-是Cl~- 吸收的竞争性抑制剂。在含200mmol L~(-1)NaCl的1/2 Hoagland培养液中,加入7.5～37.5mmolL~(-1)NO_3~-时,根系及地上部Cl~-含量降低,植株干物质积累增加.NO_3~-缓解盐胁迫的适宜浓度范围为[NO_3~-]/[Cl~-]=1/5～1/26。     

不同BR施用方式诱导黄瓜幼苗对Ca(NO_3)_2胁迫抗性的研究

为探讨外源油菜素内酯(brassinosteroid,BR)诱导黄瓜幼苗对Ca(NO3)2胁迫抗性的效果,研究了3种外源BR施用方法(0.01mg·L-1 BR浸种、0.1mg·L-1 BR喷叶及其二者结合施用)对Ca(NO3)2胁迫(60mmol·L-1)下黄瓜幼苗生长、生理活动以及光合作用的影响。结果表明:(1)3种外源BR方法处理后,Ca(NO3)2胁迫下的黄瓜幼苗株高、茎粗、展开叶片数、叶面积、干重含水量均显著提高,同时其叶片游离脯氨酸和可溶性糖含量上升,过氧化物酶活性提高,而其丙二醛(MDA)含量趋于无Ca(NO3)2胁迫对照的水平;(2)外源BR处理还提高了Ca(NO3)2胁迫下黄瓜幼苗的净光合速率、蒸腾速率和气孔导度,却抑制了Ca(NO3)2胁迫下胞间CO2浓度的升高。研究认为,适宜浓度的外源BR浸种和喷叶处理均可有效增强黄瓜幼苗渗透调节能力,降低细胞膜质过氧化伤害程度,提高抗氧化酶活性和光合效率,从而表现出对Ca(NO3)2胁迫的抗性,并以操作简便、用量极低的0.01mg·L-1 BR浸种方法效果最佳。     

铅胁迫下三叶鬼针草内源一氧化氮的生成及其对氧化损伤的缓解效应

对不同浓度铅(Pb)胁迫下三叶鬼针草(Bidens pilosa L.)叶、茎和根中内源一氧化氮(NO)和活性氧(ROS)的生成机制及根系活力的变化,内源NO对Pb胁迫下三叶鬼针草幼苗氧化损伤的缓解效应进行了研究。结果显示,在0~1000 mg/L范围内,随着Pb浓度的增加,叶片中NO含量呈升高趋势,根中NO含量呈先升高后降低的趋势,但仍高于对照,Pb浓度在0~400 mg/L范围内,茎中NO含量与对照持平,Pb浓度大于600 mg/L时,茎中NO含量低于对照;600 mg/L Pb处理能显著增强叶、茎和根中一氧化氮合成酶(NOS)和硝酸还原酶(NR)活性,显著增加叶和茎中亚硝酸根离子(NO_2~-)和类胡萝卜素(Car)含量,NOS、NR、NO_2~-和Car均能促进叶片中内源NO的生成,NOS是根中内源NO生成的主要途径。Pb胁迫使超氧阴离子(O_2~(·-))产生速率、过氧化氢(H_2O_2)含量、丙二醛(MDA)含量和相对电导率(REC)显著升高,从而造成幼苗严重的膜脂过氧化损伤,而胁迫诱发产生的NO能降低根中ROS的产生,促进幼苗根系活力,进而缓解胁迫造成的膜脂过氧化损伤。     

硝酸盐对硝酸还原酶活性的诱导及硝酸还原酶基因的克隆

硝酸盐在植物体内的积累过多已成为影响蔬菜品质并影响人类健康的重要因素。硝酸还原酶（NR）是硝酸盐代谢中的关键酶,提高其活性有利于硝酸盐的降解。为了解植物不同组织中NR的活性,用活体测定法检测了经50mmol/L的KNO₃诱导不同时间后的油菜、豌豆和番茄幼苗根茎叶中NR活性,同时为了明确外源诱导剂浓度与植物体内NR活性的关系,检测了经不同浓度KNO₃诱导2h后的矮脚黄、抗热605、小白菜和番茄叶片中的NRA。结果表明,不同植物组织NR活性有很大差异,叶中NR活性较高,根其次,茎最低;不同植物的NR活性随诱导时间呈不同的变化趋势,相同植物不同组织的NR活性变化趋势相似;不同植物叶片NRA为最高时KNO₃浓度不同。用30mmol/L的KNO3诱导番茄苗2h后,从番茄根和叶中提取总RNA,用RT-PCR方法获得NR cDNA,全长2736bp,编码911个氨基酸。为进一步利用该基因提高植物对硝酸盐的降解能力打下基础。     

Nitrate Utilization by Nitrate Reductase-deficient Barley Mutants

Two nitrate reductase-deficient barley mutants were studied for growth on nitrate and ammonium sources of nitrogen and for resistance to chlorate. Although nitrate reductase-deficient mutants in some species are chlorate-resistant (unable to reduce chlorate to chlorite), the barley mutants used in these studies when grown on nitrate and treated with chlorate were only slightly more resistant to chlorate than the control. When grown to maturity on vermiculite supplemented with either nitrate or ammonium nutrient solutions, the mutants produced as much dry weight and reduced nitrogen per plant as the control. The in vivo and in vitro nitrate reductase activities in the roots and shoots of the mutants grown on nitrate were consistently less than 10% of the control. To avoid the possibility that the mutants received reduced nitrogen from microbial sources, excised embryos were cultured under sterile conditions. Again the mutants were capable of growth and reduced nitrogen accumulation with nitrate as the sole source of nitrogen. In spite of the low apparent nitrate reductase activity, the nitrate reductase-deficient mutants are capable of substantial nitrate reduction.     

Nitrate Uptake and Assimilation following Nitrate Deprivation

Upon first exposure to , the uptake and reduction capacities of dark-grown corn (Zea maysL.) roots are initially low, but increase markedly within 6h. The development of the accelerated uptake rate appears to be substrate ‘induced’ as is reductase (NR), the first enzyme in the assimilatory pathway. However, the ‘induction’of uptake is independent of NR induction. The effect of deprivation was studied to determine the role of endogenous on subsequent uptake and reduction. Corn roots were ‘induced’ for 24 h in 0–5 mol m^–3 nutrient solution and then exposed for 0 to 32 h to -free nutrient solution. Uptake and reduction of were determined periodically by exposing sets of roots to a1 h pulse of 0.5 mol m^–3 . Neither uptake (4.57 µmol root^–1 h^–1)nor the percentage of absorbed reduced (27%) was changed significantly (P 0.05) by exogenous deprivation. However, the estimated ‘induced’ componentof uptake decreased significantly (50% after 32 h). Concurrently, the ‘non-induced’ basal componentof uptake increased. Previously accumulated decreased from 23 to 4.5 µmol root^–1 after 32 h of exogenous deprivation. Nearly equivalent quantities of endogenous were used for translocation and reduction during deprivation. During each 1 h pulse, the amounts of translocation and net efflux of to the uptake solution were similar. Net efflux of was strongly correlated (r = 0.991) to the amount of endogenous . The remaining endogenous and its rate of utilization were apparently sufficient to minimize a rapid declineor complete loss in both the ‘induced’ uptake state and the rate of in vivo assimilation. Key words: reduction, translocation, efflux, root, Zea mays L     

Stream Nitrate Responds Rapidly to Decreasing Nitrate Deposition

Ecosystem acidification and eutrophication resulting from increased deposition of dissolved inorganic nitrogen (DIN) are issues of increasing global concern. Consequently, costly policy decisions are being implemented to decrease nitrogen oxide (NO _x ) emissions. Although declining DIN deposition along with rapid declines of DIN in surface waters have been reported in parts of Europe, the same observation is just emerging in North America. Here we find a significant decline in bulk deposition NO₃ ⁻ during the later part of a 28-year record in southcentral Ontario, Canada. Despite high N retention and substantial inter-annual variability in the long-term record due to periods of drought, we find significant declines in annual NO₃ ⁻ concentrations and export at six out of 11 streams that drain upland-dominated catchments. In contrast, five streams draining primarily wetland-dominated catchments with lower levels of NO₃ ⁻ show no decreasing trend in NO₃ ⁻ concentration or export. The rapid response in stream NO₃ ⁻ to declining atmospheric inputs was observed at sites with historically moderate inputs of DIN (~870 mg m⁻² y⁻¹) in bulk deposition. Topographic features such as slope, and related catchment features including wetland cover, appear to influence which catchments will respond positively to declining DIN deposition. These findings force us to revise our original conceptualization of the N saturation status of these catchments.     

Regulation of Nitrate Assimilation and Nitrate Respiration in Aerobacter aerogenes

The influence of growth conditions on assimilatory and respiratory nitrate reduction in Aerobacter aerogenes was studied. The level of nitrate reductase activity in cells, growing in minimal medium with nitrate as the sole nitrogen source, was much lower under aerobic than anaerobic conditions. Further, the enzyme of the aerobic cultures was very sensitive to sonic disintegration, as distinct from the enzyme of anaerobic cultures. When a culture of A. aerogenes was shifted from anaerobic growth in minimal medium with nitrate and NH(4) (+) to aerobiosis in the same medium, but without NH(4) (+), the production of nitrite stopped instantaneously and the total activity of nitrate reductase decreased sharply. Moreover, there was a lag in growth of about 3 hr after such a shift. After resumption of growth, the total enzymatic activity increased again slowly and simultaneously became gradually sensitive to sonic disintegration. These findings show that oxygen inactivates the anaerobic nitrate reductase and represses its further formation; only after a de novo synthesis of nitrate reductase with an assimilatory function will growth be resumed. The enzyme in aerobic cultures was not significantly inactivated by air, only by pure oxygen. The formation of the assimilatory enzyme complex was repressed, however, by NH(4) (+), under both aerobic and anaerobic conditions. The results indicate that the formation of the assimilatory enzyme complex and that of the respiratory enzyme complex are regulated differently. We suggest that both complexes have a different composition, but that the nitrate reductase in both cases is the same protein.     

Nitrate Fluxes and Nitrate Reductase Activity of Suspension-Cultured Tobacco Cells (Effects of Internal and External Nitrate Concentrations)

Cell suspensions of tobacco (Nicotiana tabacum L., cv KY14) were used to determine the responses of NO3- uptake and NO3- reductase activity (NRA) to exogenous NO3- levels in the absence of long-distance NO3- transport. Tobacco cells grown with complete Murashige and Skoog medium for 7 d were subcultured for 3 d with NH4+-free media containing 0, 5, 10, 20, 30, and 40 mM NO3-. Cell NO3-, in vitro NRA, NO3- influx, and efflux of cell NO3- were determined. The NRA increased as cell NO3- increased. Cell NO3- efflux values increased as cell NO3- level increased. Cells with low intracellular NO3- had greater NO3- influx than cells with high intracellular NO3-. Woolf-Augustinsson-Hofstee transformations of the NO3- influx kinetic data revealed patterns characteristic of a high- and low-affinity two-component NO3- uptake system. Apparent Vmax values generally decreased and Km values increased as cell NO3- concentration increased. The NRA of cells supplied with 10 and 20 mM NO3- after 3-d growth in N- free medium increased about 5-fold within 2 h and then remained constant for the next 2 h, whereas NRA of cells supplied with 5 mM NO3- increased only 2-fold during the 4-h period. Intracellular NO3- and other N metabolites associated with cell NO3- levels exerted differential effects on the NO3- influx activity and NRA of tobacco cells cultured in suspension. Expression of high NRA was correlated with both high external and intracellular NO3-, whereas maximum NO3- influx activity required a low (depleted) level of cell NO3-.     

Nitrate Reductase mRNA Regulation in Nicotiana plumbaginifolia Nitrate Reductase-Deficient Mutants

Light and substrate regulation of nitrate reductase (NR) expression were compared in wild type and mutant lines of Nicotiana plumbaginifolia. Mutants affected in the NR structural gene (nia) or in the biosynthesis of the NR molybdenum cofactor (cnx) were examined. nia mutants expressing a defective apoenzyme, as well as cnx mutants, overexpressed NR mRNA, whereas nia mutants devoid of detectable NR protein had reduced or undetectable NR mRNA levels. Diurnal fluctuations of NR mRNA were specifically abolished in nia and cnx mutants, suggesting that the integrity of NR catalytic activity is required for the expression of diurnal oscillations. Unlike some fungal mutants, the nia and cnx mutants examined retained nitrate inducibility of NR expression. The possibility of autogenous control of NR expression in higher plants is discussed.