NO_3~-上运和液泡内NO_3~-外流对小麦叶内硝酸还原酶活性的调节

NO_3~-亏缺能使叶片硝酸还原酶活性(NRA)和NO_3~-总量降低,而根部NO_3~-吸收及上运能力提高,以亏缺2d的幼苗最为明显,该幼苗经12hNO_3~-吸收,叶片的NRA高于未经亏缺的幼苗,但NO_3~-含量以后者为高,代谢库中NO_3~-含量前者高于后者。提高营养液中NO_3~-浓度,NO_3~-上运速率升高,叶片内NRA增加。叶片组织暗中无氧保温40min后,代谢库体积渐大,液泡内NO_3~-有外流产生;Cl~-可促使液泡内NO_3~-外流,代谢库中NO_3~-量增加,NRA升高。NRA在体内测定条件下,保温3h后,NO_2~-产生趋于稳值,NRA降至最低;系统中加KCl或KNO_3使NO_2~-产生趋于稳值的时间延长,且能提高NO_2~-积累总量。     

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。     

某些环境条件对硝酸还原酶活性稳定因子的影响

小麦 Triticum aestivum L.苗在 NO_3~--N 完全营养液中培养比在 NH_4~ -N 完全营养液中培养,它们叶细胞内的硝酸还原酶(NR)即 NO_3-NR 比 NH_4-NR 活性增高了15倍,而它们叶片中的稳定因子(NR_(SF)),即 NO_3-NR_(SF)比 NH_4-NR_(SF)活化 NO_3-NR 的能力仅增加0.2倍,表明 NR 与 NR_(SF)不是依存关系;另外在 NO_3~--N 培养的黄化小麦叶片,及黄化缺氮、缺铝,加(?)的叶片中,所有的 NR_(SF)都十分稳定,并且保持较高活性,但这些叶片中没有测出NR 活性,因而认为,在植物叶细胞中,NR_(SF)不是调节 NR 活性的主要条件。     

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。     

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。     

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。     

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。     

菠菜和菜豆对二氧化氮的反应和抗性

比较了菠菜和菜豆在光下或暗中接触不同浓度NO_2时的反应。菠菜的抗性强于菜豆,是由于它既能忍受NO_2~-的大量积累,又有代谢NO_2~-的较强能力。菠菜在光照下,高浓度NO_2熏气所产生的伤害不是由于NO_2~-的积累,而是与NH_3的大量积累有关。NH_3的积累:一方面由于NiR活性不受熏气影响,另一方面由于GS/GOGAT活性受到阻抑所致。     

NaCl胁迫对大麦硝酸盐吸收和有关的酶活的影响

NaCl胁迫下,抗盐性弱的洋草麦对NO_3~-的吸收明显下降;而抗盐性强的鉴4对NO_3~-的吸收显著升高,对Cl~-的吸收则显著小于洋草麦。盐胁迫下大麦GS活力提高,GDH活力无明显变化。在低盐度下,鉴4NR活力变化较小,洋草麦则显著增加;高盐度下,两品种NR活力均迅速下降或丧失。NR活力与叶片的水势变化有显著相关。     

菠菜叶片中硝态氮还原与叶柄中硝态氮累积的关系

测定了不同生长期在不同施氮水平下3个菠菜品种各器官的硝态氮含量、叶片的硝酸还原酶活性、叶片细胞硝态氮的贮存库和代谢库大小.结果表明:叶柄中硝态氮含量远高于其它器官,其含量与叶片内源/外源硝酸还原酶活性的比值呈负相关;叶片细胞中硝态氮代谢库的大小与叶柄中硝态氮含量之间没有确定的关系.     

Critical evaluation of the in vivo nitrate reductase assay for detection of two nitrate pools in wheat leaves

The usefulness of the nitrate-free in vivo nitrate reductase assay for the study of nitrate pools in wheat leaves was investigated. Leaf sections from 7-day-old wheat seedlings, exposed 24 h before harvest to 1.5, 3.0 or 5.0 m M KNO₃ were used. After 2 to 4 h of incubation nitrite production ceased, reaching a plateau. The time required to reach the plateau and the level of the plateau increased with increasing endogenous nitrate content. At nitrite plateau the amount of nitrate left in the tissue was independent of the original nitrate content in the tissue. Addition of nitrate at plateau caused resumed nitrite production. It is concluded that nitrate was the limiting factor in nitrite production.
Oxygen inhibited nitrate reduction and stimulated further assimilation of nitrite. A considerable initial leakage of nitrate from tissue to the assay medium, followed by a slower continuous leakage, was observed throughout the incubation. N₂-flushing or inclusion of Triton X-100 in the assay medium increased nitrite production by making more nitrate available for reduction. These treatments also increased the leakage of nitrate. At plateau levels the amount of nitrate left in the tissue was dependent on the oxygen tension in the assay medium. Under low oxygen tension nearly all nitrate in the tissue was available for reduction. Nitrite production at plateau is not a useful index for a metabolic nitrate pool and nitrate left in the tissue is not a useful index for a nitrate storage pool because both parameters are highly dependent on the oxygen tension in the assay medium. Further, in view of the considerable leakage, the nitrate-free in vivo nitrate reductase assay cannot be used to detect two separate nitrate pools in wheat leaves.     

不同蛋白质含量小麦品种叶片NRA与氮素积累关系的研究

以鲁麦５号和昌乐５号两种不同蛋白质含量小麦品种为材料,研究了各生育期叶片ＮＲＡ与氮素积累的关系。其结果是,在不同施氮量下,各生育期叶片ＮＲＡ、ＮＯ３＾－－Ｎ、ＮＨ２－Ｎ、还原Ｎ含量皆随施氮量增加而增加,但生育前期昌乐５号的毕大于鲁麦５号,而生育后期则相反,鲁麦５号的皆大于昌乐５号,籽粒蛋白质含量亦为鲁麦５号高于昌乐５号。表明生育后期（开花期后）叶片ＮＲＡ是反映籽粒蛋白质含量高低的一项重要指标。     

Some New Aspects of the in Vivo Assay for Nitrate Reductase in Wheat (Triticum aestivum L.) Leaves: I. REEVALUATION OF NITRATE POOL SIZES

Experiments were carried out to clarify problems encountered in measuring metabolic and storage pool sizes of nitrate in wheat leaf sections with an in vivo nitrate reductase assay. The leaf sections were from seedlings grown on 15 millimolar nitrate. Data obtained show that the cessation of nitrite accumulation, used as an index of the active nitrate pool size, could be caused by lack of anaerobiosis in the assay system, the lack of energy for nitrate reduction, or a loss of nitrate reductase activity. Availability of nitrate was never the limiting factor in this system. It is concluded that pool sizes of nitrate cannot be determined in wheat leaves with the in vivo assays employed.     

The effect of cytokinins on nitrate reductase activity

Cytokinins in addition to nitrate induce nitrate reductase activity (NRA) in some plants. Effects of cytokinins onNRA was investigated in stem pith parenchyma of kale, intact wheat and barley seedlings and isolated cucumber cotyledons. The most profound effect onNRA was found in barley and wheat seedlings.NRA in seedlings sprayed with 100 μM 6-benzylaminopurine (BAP) for three subsequent days was increased in leaves and decreased in roots. These changes were further enhanced in seedlings grown in nutrient solution lacking nitrate:NRA in wheat and barley leaves was increased by 57% and 202%, respectively, in plants supplied with nitrate theNRA increase was not significant: in wheat and barley leaves by 22% and 9%, respectively. Similar effect of BAP and kinetin was found in kale stem parenchyma and cucumber cotyledons. The cytokinin kinetin or BAP alone increasedNRA about twice in kale and three times in cucumber. Addition of nitrate to the medium enhanced the effect of kinetin in kale discs, but the two effects were not additive. Additive effect of nitrate and BAP onNRA was found in cucumber cotyledons in light. In general NRA was more affected by cytokinins in intact seedlings of wheat and barley as compared to explanted tissue of kale and cucumber, and lack of nitrogen made their effect more expressive.     

Nitrate Reductase Activity in Maize (Zea mays L.) Leaves: II. Regulation by Nitrate Flux at Low Leaf Water Potential

Experiments were conducted to determine whether the nitrate flux to the leaves or the nitrate content of the leaves regulated the nitrate reductase activity (NRA) in leaves of intact maize (Zea mays L.) seedlings having low water potentials (ψ_w) when other environmental and endogenous factors were constant. In seedlings that were desiccated slowly, the nitrate flux, leaf nitrate content, and NRA decreased as ψ_w decreased. The decrease in nitrate flux was caused by a decrease in both the rate of transpiration and the rate of nitrate delivery to the transpiration stream. Upon rewatering, the recovery in NRA was correlated with the nitrate flux but not the leaf nitrate content.     

Time-course of Nitrate Uptake and Nitrate Reductase Activity in Nitrogen-depleted Dwarf Bean

The rate of nitrate uptake by N-depleted French dwarf bean (Phaseolus vulgaris L. cv. Witte Krombek) increased steadily during the first 6 h after addition of NO₃ -After this initial phase the rale remained constant for many hours. Detached root systems showed the same time-course of uptake as roots of intact plants. In vivo nitrate reductase activity (NRA) was assayed with or without exogenous NO₃^- in the incubation medium and the result ing activities were denoted potential and actual level, respectively. In roots the difference between actual and potential NRA disappeared within 15 min after addition of nitrate, and NRA increased for about 15 h. Both potential and actual NRA were initially very low. In leaves, however, potential NRA was initially very high and was not affected by ambient nitrate (0.1–5 mol m^-3) for about 10 h. Actual and potential leaf NRA became equal after the same period of time. In the course of nitrate nutrition, the two nitrate reductase activities in leaves were differentially inhibited by cycloheximide (3.6 mmol m^-3) and tungstate (1 mol m^-3). We suggest that initial potential NRA reflects the activity of pre-existing enzyme, whereas actual NRA depends on enzyme assembly during NO₃^- supply. Apparent induction of nitrate uptake and most (85%) of the actual in vivo NRA occurred in the root system during the first 6 h of nitrate utilization by dwarf bean.     

NO2‐induced nitrate reductase activity in needles of Norway spruce (Picea abies) under laboratory and field conditions

The induction of activity of the enzyme nitrate reductase (NR, EC 1.6.6.1, 1.6.6.2) in needles of Norway spruce ( Picea abies [L.] Karst.) by nitrogen dioxide (NO₂) was studied under laboratory and field conditions. In fumigation chambers an increase in nitrate reductase activity (NRA) was detected 4 h after the start of the NO₂ treatment. During the first 2 days with 100 µg NO₂ m⁻³, NRA reached a constant level and did not change during the following 4 days. At the same level of NO₂, NRA was lower in needles from trees grown on NPK‐fertilized soil than on non‐fertilized soil. After the transfer of spruce trees from fertilized soil to NPK‐rich nutrient solution, NRA was transiently increased. This effect was assigned to root injuries causing nitrate transport to the shoot and subsequent induction of NRA. Neither trees on fertilized soil nor trees transferred to NPK‐poor nutrient solution had increased NRA unless NO₂ was provided. The NO₂ gradient in the vicinity of a highway was used to test the long‐term effect of elevated levels of NO₂ on needle NRA of potted and field‐grown spruce trees. Compared with less polluted sites, permanently increased NRAs were detected when NO₂ concentrations were above 20 µg m⁻³. Controls of field measurements some 10 years after the introduction of catalytic converters in cars showed no significant change neither in NO₂ levels nor in the decreasing NRA of spruce needles with the distance from the highway.     

Reevaluation of anaerobic nitrite production as an index for the measurement of metabolic pool of nitrate

The use of anaerobic nitrite production as an index for the measurement of metabolic pool of nitrate was reevaluated using primary leaves of 7-day-old barley and 10-day-old soybean seedlings. The seedlings were grown in nutrient solutions containing 5 to 15 millimolar nitrate. The nitrate-free in vivo assay system of nitrate reductase was used for measuring the production of nitrite. Both the duration and extent of nitrite production were dependent on the level of endogenous nitrate in the tissue. At cessation of nitrite production, 30 to 50% of the endogenous nitrate was reduced to nitrite. Nitrate from the tissue leaked continuously into the surrounding medium so that, at cessation of nitrite production, nitrate supply from the tissue was exhausted. The cessation of nitrite production, therefore, may have been caused by the depletion of endogenous nitrate from the tissue. It is concluded that anaerobic nitrite production is not a valid index for the measurement of the size of the metabolic pool of nitrate.