硝酸还原酶抑制剂钨酸钠对油菜硝态氮积累的影响

采用溶液培养方法,选取硝酸盐积累差异明显的两个油菜品种（低硝态氮积累品种‘红油3号’和高硝态氮积累品种‘中双6号’,研究苗期根系硝酸还原酶（NR）活性被抑制以后两个油菜品种叶片、叶柄和根系中NR活性和硝态氮含量的变化。结果表明：1.0mmol.L-1的NR活性抑制剂Na2WO4对两个油菜品种的根系NR活性抑制效果最佳;根系NR活性被抑制以后,两个油菜品种的根系NR活性、硝态氮吸收速率均显著下降,而硝态氮含量却显著上升;且Na2WO4对‘中双6号’硝态氮吸收的抑制程度强于其对‘红油3号’的抑制。叶片和叶柄的NR活性变化不显著,但叶柄硝态氮含量显著下降,叶片硝态氮含量稳定,且这一趋势在低积累品种‘红油3号’中表现得更为明显。     

大豆植株发育过程中不同部位的硝态氮含量和硝酸还原酶活力的变化

同一时期的大豆品种长农5号植株中硝态氮含量是,主茎大于叶柄大于叶片,前二者中硝态氮含量随着生育进程逐渐减少,生育前期由基部向上逐渐增加,鼓粒期以后各节位间无明显变化。营养生长期间,叶片中硝态氮含量较高,不同生殖生长期间或同一时期的不同节位变化不明显。硝酸还原酶活力是成熟叶片中低,幼嫩叶片中高,且随着生育进程逐渐降低。     

不同品种菠菜叶肉及叶柄中硝态氮累积与硝酸还原酶活性的关系

盆栽试验条件下,菠菜体内的硝态氮含量及硝酸还原酶（NR）活性因品种和部位而异。硝态氮含量以短缩茎中最高,叶和叶柄中次之,根中最低,且根,冠比越大的品种,其整体植株中的硝态氮含量越低;根尤其是侧根NR活性较高;叶肉和叶柄中硝态氮含量均与根中以内、外源硝酸盐为底物的NR活性呈一定的负相关。     

菠菜叶片中硝态氮代谢库的测定（简报）

随着培养时间的延长 ,菠菜组织中还原产生的亚硝态氮量呈波动型上升 ;以 pH 7.5的磷酸缓冲液为培养介质的亚硝态氮累积量显著高于以KCl和CaCl2 混合溶液 ( pH 6.65 )为介质的 ;培养前不通氮气 ,培养后期亚硝态氮的生成量明显高于通氮气的。亚硝态氮生成第一峰值出现的时间随硝态氮含量增高而后延 ,峰值却随硝态氮含量的增高而升高。用 pH 7.5的 0 .0 5mol·L-1磷酸缓冲液且不通氮气对组织进行培养 ,亚硝态氮生成的第一个峰值代表硝态氮代谢库的大小。     

硝态氮和铵态氮对墨兰生长发育的影响

以无土栽培的方法研究不同氮水平的硝态氮和铵态氮对墨兰［Ｃｙｍｂｉｄｉｕｍｓｉｎｅｎｓｅ（Ａｎｄｒ．）Ｗｉｌｌｄ．］生长发育和某些生理特性的影响。１ｍｍｏｌ／Ｌ和１０ｍｍｏｌ／Ｌ的硝态氮和铵态氮处理都促进叶芽和叶片正常生长,前者似乎比后者好一些。５０ｍｍｏｌ／Ｌ的两种形态的氮均使叶片生长缓慢,尤其是铵态氮。１和１０ｍｍｏｌ／Ｌ硝态氮处理则正常开花,而５０ｍｍｏｌ／Ｌ则例外。铵态氮处理各种浓度均不形成花芽。建议墨兰营养生长期施用１ｍｍｏｌ／Ｌ的硝态氮或铵态氮,但生长后期则应施１—１０ｍｍｏｌ／Ｌ硝态氮,以利花芽分化。作者认为,墨兰生长缓慢与光合速率极慢和硝酸还原酶活性极低有关,墨兰开花与否和Ｅ／Ｎ比大小有关。     

施氮对不同品种冬小麦植株硝态氮和硝酸还原酶活性的影响

以黄土高原南部半湿润区土垫旱耕人为土为供试土壤进行盆栽试验,以NR 9405、9430、偃师9号、小偃6号、陕229号和西农2208冬小麦品种为供试材料,研究施氮对不同品种冬小麦植株硝态氮含量和硝酸还原酶活性(NRA)的影响.结果表明,施氮能明显增加叶片NRA.不施氮时除小偃6号和偃师9号外,其余品种NRA在全生育时期的动态变化均呈双峰曲线,2个高峰期分别在返青期和开花期,且开花期高峰值(36.17 NO2-μg.-g 1FW.h-1)明显比返青期峰值(15.407 NO2-μg.-g 1FW.h-1)大;施氮时不同品种叶片NRA在全生育期呈单峰曲线变化,最高峰在开花期,平均峰值为80.93 NO2-μg.-g 1FW.h-1),比同期不施氮处理增加1倍以上.施氮后地上部硝态氮含量在各时期均显著提高,在小麦生育前期(出苗到拔节)表现最为显著.氮肥对不同品种硝态氮含量的影响程度基本上与对NRA的影响程度相反,即施氮后硝态氮增加幅度小的品种,NRA却增加幅度大.     

硝态氮对小白菜铬污染毒性的调控作用

通过盆栽和水培试验,探讨了硝态氮对小白菜铬污染毒性的调控作用。结果表明:外源Cr6 对小白菜铬吸收和积累具有明显的促进效应,抑制了小白菜对铁养分的吸收并降低了小白菜的硝酸还原酶活性;硝态氮可有效缓减Cr6 对小白菜吸收铁和硝酸还原酶活性的抑制作用,促进小白菜碳氮代谢和Vc的生物合成,并刺激小白菜生长。在相同铬污染条件下,土壤硝态氮的增加促进了外源Cr6 向有机态转化,且硝态氮有协同强化小白菜吸收Cr6 的效应。表明硝态氮在促进小白菜生长的同时,也促进了小白菜对Cr6 的吸收,提高了小白菜的铬累积水平。     

叶菜类蔬菜不同部位的硝态氮累积对氮肥的响应

土壤盆栽油菜、小白菜和菠菜的叶柄对施氮的响应最为敏感。叶柄中硝态氮的累积量占整株蔬菜的一半以上（从54,9％到75．0％）。叶柄中硝态氮累积量与植物整株的硝态氮累积量呈极显著正相关。     

不同浓度硝态氮供应下小麦生长、硝态氮累积及根系钙信号特征

以小麦品种‘石麦15’和‘衡观35’为材料进行营养液水培试验,研究不同浓度硝态氮供应对小麦苗期根系形态、钙离子流特征及钙调蛋白(CaM)含量的影响。结果表明,与适宜浓度硝态氮处理(2.5mmol/L)相比,无外源硝态氮供应时小麦地上部鲜重、硝态氮含量均降低,侧根数量显著减少;高浓度硝态氮处理(50mmol/L)下两个小麦品种地上部硝态氮含量升高,根系总长度降低,‘石麦15’侧根数量减少。无硝态氮和高浓度硝态氮处理下,根系中钙调蛋白含量降低,且‘衡观35’的降低幅度大于‘石麦15’。无外源硝态氮供应时小麦根尖表现出较为明显的钙离子外流特征;与适宜浓度硝态氮处理相比,高硝态氮处理下小麦根尖Ca2+的内流速度显著下降。说明硝态氮供应不足和高浓度硝态氮供应会影响小麦根系生长,根系Ca2+外流或Ca2+内流速度下降,CaM含量减少,Ca2+/CaM可能介导硝态氮调控小麦根系生长发育。     

叶类蔬菜的硝态氮累积及成因研究

在菜园土壤上进行的田间试验,用禾谷类作物冬小麦作比较,研究了菠菜、小白菜、大青菜和油菜等叶类蔬菜累积硝态氮的特点,结果表明：硝态氮累积是一般早作作物的共性,苗期更为明显,无论蔬菜还是冬小麦均有较高的硝态氮含量（367.8-1413.4μg/g);但随生育期后延,蔬菜的硝态氮含量波动升高,冬小麦波动降低,盆栽试验表明,施入土壤的氮肥是蔬菜硝态氮累积的主要来源,过量施用氮肥所导致的蔬菜硝态氮吸收与还原转化不平衡是产生累积的根本原因,吸收与生长不协调更使累积过程加剧。     

The Mechanism of Nitrate Accumulation in Pakchoi [Brassica Campestris L.ssp. Chinensis(L.)]

Decreased nitrate in vegetables can improve crop nitrogen utilization efficiency and lessen the human health risk caused by the reduction of nitrate to nitrite in vegetables. This paper studied the mechanisms of differences in nitrate accumulation and distribution within organs of two cultivars of pakchoi (Brassica campestris L.ssp. Chinensis (L.) previously screened in hydroponic experiments from 12 cultivars popularly grown in China at present. The two typical cultivars used in this experiment were Shanghaiqing with low nitrate accumulation and Liangbaiye 1 with high nitrate accumulation. There was no significant difference of total nitrate uptake but a significant difference in nitrate content existed between the two cultivars. Compared with Liangbaiye 1, Shanghaiqing showed a significantly higher photosynthetic rate and nitrate reductase activity. Determination of nitrate concentration (activity) in vacuoles with double-barrelled nitrate-selective microelectrodes showed that Shanghaiqing had lower vacuolar nitrate activity than Liangbaiye 1. Two putative nitrate reductase genes, nia1 and nia2, were amplified from the leaf blades of these two cultivars. Nia1 mRNA fragments (887 bp, accession numbers DQ082868 and DQ082869) were amplified using degenerate primer and nia2 mRNA fragment was amplified using one pair of generate primers designed according to DQ001901. Sequence analysis of DQ082868 and DQ082869 both showed 97% and 87% similarity with two nitrate reductase mRNA sequences of Brassica napus, accession numbers D38219 and D38220, respectively. The results of real time PCR to compare the relative expression of the putative nitrate reductase genes (nia1 and nia2) showed that Shanghaiqing had significantly higher expression level than Liangbaiye 1 and nia2 was significantly higher than nia1 in leaf blade and petiole. Both the nitrate reductase activity and the relative expression level of nia1 were in the order of leaf blade > root > petiole, while that of nia2 was leaf blade > petiole > root. There was no statistically significant difference of nitrate activity stored in vacuoles between the different organs of the two cultivars. It can be concluded that Shanghaiqing took up slightly less nitrate, but had significantly higher nitrate reductase activity in cytosol and had a higher relative expression of the putative nitrate reductase genes than Liangbaiye 1; this leads to the fact that Shanghaiqing has a lower nitrate content than Liangbaiye 1.     

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。     

Effect of one night with "low light'on uptake, reduction and storage of nitrate in spinach

During the night, shoot nitrate concentration in spinach (Spinacia oleracea L. cv. Vroeg Reuzenblad) increased due to increased uptake of nitrate by the roots. When the plants were subjected to a one night “low light’period at 35 μmol m^?2 s^?1, the shoot nitrate concentration did not increase and was reduced by 25% compared to control plants in the dark. The major contribution to this decrease was located in the leaf blades, where the nitrate concentration was decreased by 60%, while the petiole nitrate concentration decreased by only 9%. Nitrate accumulated in the leaf blade vacuoles during a dark night, but this was not the case during the “low light’period. This decrease in vacuolar nitrate concentration, compared to control plants in the dark, was not caused by increased amounts of leaf blade nitrate reductase (NR; EC 1.6.6.1). During a “low light’night period, the cytoplasmic soluble carbohydrate concentration was increased compared to the control plants in the dark. Calculations showed in situ NR activity to be higher than in the control plants in the dark. This increase in NR activity, however, was not large enough to account for the total difference found in the shoot nitrate concentration. Net uptake of nitrate by the roots was increased during the initial hours of the dark night, while vacuolar nitrate concentration in the leaf blades increased at the same time. During the “low light’night period, however, net uptake of nitrate by the roots did not increase, and vacuolar nitrate concentration did not change. We conclude that nitrate uptake by the roots and vacuolar nitrate concentration in the leaf blades are tightly coupled. The decreased shoot nitrate concentration is mainly caused by a reduction in net uptake of nitrate by the roots. During the “low light’night period, carbohydrates and malic acid partly replaced vacuolar nitrate. A “low light’period one night prior to harvest provides a valuable tool to reduce shoot nitrate concentrations in spinach grown in greenhouses in the winter months.     

Nitrate metabolism in roots and nodules of Vicia faba in response to exogenous nitrate

Activities of nitrate reduction enzymes, nitrate reductase activity (NRA) and nitrite reductase activity (NiRA) from roots and nodules of 5 mutant genotypes and one commercial cultivar (Alameda) of faba bean ( Vicia faba L. var. minor) grown in the presence of N₂ alone or with additional NO⁻₃ in the medium have been studied. A naturally occurring mutant (VFM109) with impaired ability to reduce nitrate in its nodules is described. All the other cultivars of V. faba showed nodule NRA, although the range was very wide, from almost negligible (VFM72) up to 2 μmol h⁻¹ (g FW)⁻¹. This activity was entirely of plant origin. Root NRA also ranged widely accross cultivars. However, the level of activity expressed as well as the response of NRA to nitrate followed a pattern opposite to that observed in nodules. Roots and nodules of all cultivars showed very high rates of NiRA, respectively 50 and 150-fold higher than NRA, thus precluding accumulation of nitrite in these tissues. Root enzymes were significantly stimulated by nitrate while negative (NRA) or little effect (NiRA) was found for nodules. Nitrate and nitrite reduction are carried out by inducible enzymes in roots of V. faba and by constitutive enzymes in nodules, indicating that there may be different forms of these enzymes in each tissue. Differences in the plant genotype were a major cause of the variability in nitrate and nitrite reduction by nodulated root systems of V. faba .     

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 reductase in cotyledons of cucumber seedlings as affected by nitrate, phytochrome and calcium

Regulation by the active form of phytochrome (P_FR) and the effect of Ca²⁺ was examined with nitrate reductase (NR) in etiolated cucumber ( Cucumis sativus cv. Beilpuig). Nitrate reductase activity (NRA) was studied in excised cotyledons of cucumber seedlings grown in distilled water and in darkness for seven days at 24 ± 0.5°C. All experiments were performed in the dark and a dim green safelight was used during analyses. In etiolated cucumber cotyledons NRA was induced by nitrate and a brief irradiation (15 min) with red light (R) resulted in 62% increase in NRA. This effect was nullified when R was followed immediately by a brief (5 min) far-red light (FR). NRA also showed a semidian (12 h) rhythmicity. Both P_FR, and nitrate effects were age dependent. Calcium seemed to be involved since the phytochrome effect was only observed when calcium was supplied in the external solution. The effect of R on NRA depended on the period of calcium nitrate incubation. An external supply of calcium ionophore mimicked the effect of R and, if supplied to R-irradiated cotyledons, produced a higher NR level than that caused by R alone. This suggested that intracellular free calcium was involved.     

Nitrate uptake,nitrate reductase distribution and their relation to proton release in five nodulated grain legumes

Nitrate uptake, nitrate reductase activity (NRA) and net proton release were compared in five grain legumes grown at 0.2 and 2 mM nitrate in nutrient solution. Nitrate treatments, imposed on 22-d-old, fully nodulated plants, lasted for 21 d. Increasing nitrate supply did not significantly influence the growth of any of the species during the treatment, but yellow lupin (Lupinus luteus) had a higher growth rate than the other species examined. At 0.2 mM nitrate supply, nitrate uptake rates ranged from 0.6 to 1.5 mg N g(-1) d(-1) in the order: yellow lupin > field pea (Pisum sativum) > chickpea (Cicer arietinum) > narrow-leafed lupin (L angustifolius) > white lupin (L albus). At 2 mM nitrate supply, nitrate uptake ranged from 1.7 to 8.2 mg N g(-1) d(-1) in the order: field pea > chickpea > white lupin > yellow lupin > narrow-leafed lupin. Nitrate reductase activity increased with increased nitrate supply, with the majority of NRA being present in shoots. Field pea and chickpea had much higher shoot NRA than the three lupin species. When 0.2 mM nitrate was supplied, narrow-leafed lupinreleased the most H+ per unit root biomass per day, followed by yellow lupin, white lupin, field pea and chickpea. At 2 mM nitrate, narrow-leafed lupin and yellow lupin showed net proton release, whereas the other species, especially field pea, showed net OH- release. Irrespective of legume species and nitrate supply, proton release was negatively correlated with nitrate uptake and NRA in shoots, but not with NRA in roots.     

Effects of soil applied herbicides on leaf nitrate reductase and crude protein in the leaf and seed of two cowpea cultivars

A glass-house study was conducted to determine the effects of four commonly used herbicides (pendimethalin, metobromuron, metolachlor and prometryne) applied pre-emergence at rates of 0, 0.125, 0.625 and 1.25 kg ha^–1, on leaf nitrate concentration (NO₃–C), nitrate reductase activity (NRA), leaf crude protein and seed protein in two cowpea cultivars, 60 day (60D) and Ife brown (IB).Control and treated plants of both cultivars showed separate peaks for NO₃–C and NRA, 49 days after planting (DAP) and 35 DAP for 60D and IB respectively. Herbicide treatment generally enhanced NO₃–C but tended to decrease NRA in both cultivars. Howver, metobromuron at 0.625 kg ha^–1 increased NRA throughout the growth period with an optimum increase of 52.5%, over the control, at 35 DAP. Pendimethalin increased NO₃–C NRA and leaf protein but did not influence seed protein appreciably. In contrast metobromuron increased NO₃–C, decreased NRA, but increased seed protein by 29.6% over the control at 0.125 kg ha^–1 in 60D. Metolachlor and prometryne were most inhibitory to seed protein development. In addition, metolachlor reversed the interdependence of NO₃–C and NRA.     

Effect of amino compounds on nitrate utilization by roots of dwarf bean

Amino compounds (1 mM, pH 5) were given prior to, together with, or after the addition of nitrate to study their effect on nitrate uptake and in vivo nitrate reductase activity (NRA) in roots of Phaseolus vulgaris. The effect of amino compounds varied with the amino species, the nitrate status of the plant (induced vs uninduced) and the aspect of nitrate utilization. Cysteine inhibited the nitrate uptake rate and root NRA under all conditions tested. NRA in uninduced roots was stimulated by tryptophan, and arginine inhibited NRA under all conditions tested. Uptake was inhibited by aspartate and glutamate and stimulated by leucine when these amino compounds were given prior to or after completion of the apparent induction of nitrate uptake. In the presence of β-alanine and tryptophan, induction of uptake was accelerated.     

Latent nitrate reductase activity is associated with the plasma membrane of corn roots

Latent nitrate reductase activity (NRA) was detected in corn (Zea mays L., Golden Jubilee) root microsome fractions. Microsome-associated NRA was stimulated up to 20-fold by Triton X-100 (octylphenoxy polyethoxyethanol) whereas soluble NRA was only increased up to 1.2-fold. Microsome-associated NRA represented up to 19% of the total root NRA. Analysis of microsomal fractions by aqueous two-phase partitioning showed that the membrane-associated NRA was localized in the second upper phase (U₂). Analysis with marker enzymes indicated that the U₂ fraction was plasma membrane (PM). The PM-associated NRA was not removed by washing vesicles with up to 1.0 M NACl but was solubilized from the PM with 0.05% Triton X-100. In contrast, vanadate-sensitive ATPase activity was not solubilized from the PM by treatment with 0.1% Triton X-100. The results show that a protein capable of reducing nitrate is embedded in the hydrophobic region of the PM of corn roots.Abbreviations L₁ first lower phase - NR nitrate reductase - NRA nitrate-reductase activity - PM plasma membrane - T:p Triton X-100 (octylphenoxy polyethoxyethanol) to protein ratio - U₂ second upper phase