Nitrate Reductase Activity In Wheat Research Articles

Nickel toxicity mitigation by supplementation of acetylcholine in wheat: Growth, photosynthetic and antioxidant activities

Influence of exogenous treatment of acetylcholine (ACh; 5 and 10 µM) was studied on the growth, photosynthesis and nitrate reductase activity in wheat under nickel (500 µM Ni) stress. Plant height, fresh and dry weight of shoot showed a decline of 36%, 23% and 40% respectively while as increased by 35%, 44% and 47% due to 10 µM ACh over control. Treatment of ACh, at both concentrations, alleviated the decline with obvious effects at higher (10 µM) concentrations. Treatment of ACh resulted in increased content of total chlorophylls, carotenoids and photosynthesis, intercellular carbon dioxide, transpiration and PSII activity, which were however significantly reduced by Ni stress. Exogenous application of ACh alleviated the Ni stress mediated decline in the activity of nitrate reductase. Wheat plants treated with Ni showed increased production of hydrogen peroxide resulting in increased lipid peroxidation and electrolyte leakage, however treatment of ACh reduced these parameters significantly reflecting in alleviation of oxidative damage. Activity of antioxidant enzymes assayed (SOD, CAT, APX, DHAR and GR) and the content of ascorbic acid and reduced glutathione increased significantly in ACh treated plants under normal and Ni stress. Activities of SOD, CAT, APX, DHAR and GR was increased by 162%, 108%, 70%, 107% and 87% respectively in Ni + 10 µM ACh treated plants. In addition, the content of phenols and flavonoids increased due to application of ACh. Content of nitrogen, potassium and calcium increased due to ACh and also alleviated the decline induced due to Ni stress while as the content of Ni was reduced in plants treated with ACh.

Effects of warming and elevated O3 concentrations on N2O emission and soil nitrification and denitrification rates in a wheat-soybean rotation cropland

The effects of warming and elevated ozone (O3) concentrations on nitrous oxide (N2O) emission from cropland has received increasing attention; however, the small number of studies on this topic impedes understanding. A field experiment was performed to explore the role of warming and elevated O3 concentrations on N2O emission from wheat-soybean rotation cropland from 2012 to 2013 using open-top chambers (OTCs). Experimental treatments included ambient temperature (control), elevated temperature (+2 °C), elevated O3 (100 ppb), and combined elevated temperature (+2 °C) and O3 (100 ppb). Results demonstrate that warming significantly increased the accumulative amount of N2O (AAN) emitted from the soil-winter wheat system due to enhanced nitrification rates in the wheat farmland and nitrate reductase activity in wheat leaves. However, elevated O3 concentrations significantly decreased AAN emission from the soil-soybean system owing to reduced nitrification rates in the soybean farmland. The combined treatment of warming and elevated O3 inhibited the emission of N2O from the soybean farmland. Additionally, both the warming and combined treatments significantly increased soil nitrification rates in winter wheat and soybean croplands and decreased denitrification rates in the winter wheat cropping system. Our results suggest that global warming and elevated O3 concentrations will strongly affect N2O emission from wheat-soybean rotation croplands.

Selenium nano-particle induced alterations in expression patterns of heat shock factor A4A (HSFA4A), and high molecular weight glutenin subunit 1Bx (Glu-1Bx) and enhanced nitrate reductase activity in wheat (Triticum aestivum L.)

Taking account of heat shock factor A4A (HSFA4A) as a hydrogen peroxide sensor, anti-apoptosis agent, and crosslink component with critical signaling cascades, the current study was carried out to monitor possible changes in expression of this gene as well as some other important characteristics in wheat plants exposed to selenium nano-particle (nSe). Wheat seedlings were treated with nSe (0, 5, 10, and 50 mgl−1). In germinating stage, plant fresh weights were reduced in nSe-treated seedlings, among which the nSe of 50 provoked roots turned brown. The nSe triggered the increases in the expressions of HSFA4A, in the plate. In the pot condition, shoot fresh weights in nSe-supplemented seedlings were decreased by approximately 22%. The nSe of 5 and 10 mgl−1, respectively induced the expression of HSFA4A by 3.4- and 9.15-folds, contrasted with nSe50. Increasing times of sprays caused the dramatic reductions of the expression of HSFA4A in the nSe-supplemented groups. Moreover, the treatment of nSe stimulated expression of high molecular weight glutenin subunit 1Bx (Glu-1Bx) by fourfold, over the control. While the dramatic decrease in the expression of Glu-1Bx was recorded with increasing times of spray. Nitrate reductase activities were significantly improved by approximately 47% in nSe-fortified seedlings. Also, the foliar supplementation of nSe of 5 mgl−1 provoked the significant inductions in peroxidase activity by 8%, whereas two other nSe treatments declined it. It may be stated that the nSe may modify the expression of HSFA4A, thereby triggering specific signaling and altering metabolism.

Nitric Oxide Mediates Molybdenum-Induced Antioxidant Defense in Wheat under Drought Stress.

Molybdenum (Mo) has been reported to alleviate drought stress by enhancing antioxidant defense in plants, but the underlying mechanism remains unclear. Here, we hypothesized that Mo mediates nitric oxide (NO)-induced antioxidant defense through Mo-enzymes, particularly by nitrate reductase (NR) in wheat under drought stress. The 30-day-old wheat seedlings cultivated in -Mo (0 μM Mo) and +Mo (1 μM Mo) Hoagland solutions were detached and then pretreated with Mo-enzyme inhibitors, NO scavengers, NO donors or their combinations according to demands of complementary experiment under 10% polyethylene glycol 6000 (PEG)-stimulated drought stress (PSD). Mo supplementation increased the activities and transcripts of antioxidant enzymes, decreased H2O2 and MDA contents, and elevated NO production, implying that Mo-induced antioxidant defense may be related to NO signal. Complementary experiment showed that NO production was induced by Mo, while suppressed by Mo-enzyme inhibitors and NO scavengers, but restored by NO donors, suggesting that Mo-induced increase of NO production may be due to the regulation by Mo-enzymes. Further experiment indicated that the increased activities and transcripts of antioxidant enzymes induced by Mo were suppressed by Mo-enzyme inhibitors and NO scavengers, and NO donors could eliminate their suppressing effects. Moreover, Mo application increased NR activity and inhibitors of Mo-enzymes inhibited NR activity in wheat leaves under PSD, suggesting that NR might involve in the regulation of Mo-induced NO production. These results clearly indicate that NO mediates Mo-induced antioxidant defense at least partially through the regulation of NR.

The dependence of nitrate reductase activity on the level of soluble sugars in wheat and cucumber roots growing in the presence of simazine, in light or in darkness

At concentrations of 25 and 5 µM, simazine inhibited nitrate reductase activity in wheat and cucumber roots, respectively. It also lowered the content of soluble sugars and decreased the activities of NADH malate dehydrogenase and NADP<sup>+</sup> glucose-6-phosphate dehydrogenase. The inclusion of 50 mM glucose into the medium partially reversed the inhibitory effect of simazine on the activity of nitrate reductase in cucumber roots and slightly increased the activity of this enzyme in wheat roots These results suggest a complex influence of the herbicide on the activity of nitrate reductase: simazine lowers the level of soluble sugars in roots and decreases the activity of the dehydrogenases supplying the reduced nucleotides indispensable for reduction of nitrates.

Nitric Oxide Content in Wheat Leaves and Its Relation to Programmed Cell Death of Main Stem and Tillers Under Different Nitrogen Levels

Nitric oxide (NO) is a key signaling molecule in different physiological processes of plants, including programmed cell death (PCD). PCD of tillers plays an important role in surviving which are major components of grain yield. PCD was triggered in wheat leaves of main stem and tillers by NO content under different nitrogen treatments. In wheat, NO could be synthesized endogenously by nitrate reductase (NR). As an inducible enzyme, NR activity was closely related to substrate concentration. Therefore, different nitrogen levels would change NR activity and NO production. The objective of this study was to determine the effects of NR activity, NO production, and the correlation between them on different tillers growth, development, senescence, and kernel protein content under different nitrogen levels. Field-experiments were conducted in 2009–2011 growing seasons, using two wheat cultivars with different spike-types. Results showed that for main stem and primary tillers, NR activity and NO content reached high level at heading stage, while for secondary tiller, the level of NR activity was low, but NO content was high in the present research. The NO synthesis depending on NR activity in wheat leaves was significant in the early growing stage, but the NO synthesis weakened with the progress of growing period. NO was related to the senescence of wheat leaves, but PCD was more sensitive to marked changes of NO content than NO content itself. N application had marked influence on the aging process of primary tiller, while had little influence on that of main stem and secondary tiller. Moreover, N fertilizer application could increase spike rate and protein content of primary tiller by N fertilizer application.

Nitrate reductase from Triticum aestivum leaves: Regulation of activity and possible role in production of nitric oxide

Nitrate reductase (NR) and peroxidase (POX) are important enzymes involved in the metabolism of reactive oxygen (ROS) and nitrogen species in leaves of wheat (Triticum aestivum L.) seedlings. It has been confirmed that NR activity in wheat leaves depends on the light conditions and the presence of nitrates during the cultivation of the seedlings, and it is regulated by the molybdenum cofactor and phosphorylation. In the present study, confocal microscopy and EPR spectroscopy studies showed that the addition of nitrite, a product of NR, increased the level of nitric oxide (NO). This increase was prevented by the addition of sodium azide, an inhibitor of NR. The results suggest that in wheat leaves one of the key functions of NR is the formation of the signaling NO molecule. Cultivation of green plants under conditions of prolonged (4 days) darkness, a strong stress factor for photosynthesizing cells, decreased the activity of NR. Moreover, darkness induced significant elevation of the POX activity that was prevented by the addition of nitrate to the growth medium. It is proposed that the changes in light conditions result in the competition between nitrate- and ROS-metabolizing activities of POX in leaves, and a possible interaction between NR and POX controls the levels of NO and ROS in the leaf tissue.

Polyamines modulate nitrate reductase activity in wheat leaves: involvement of nitric oxide

In the present work, the effect of polyamines (PAs) on nitrate reductase (NR) activity was studied in wheat leaves exposed to exogenously added PAs while assessing the nitric oxide (NO) involvement in the regulation of the enzyme activity. A biphasic response was observed along the time of treatment using 0.1 mM of putrescine (Put), spermidine (Spd) or spermine (Spm). At 3 h, Spd and Spm significantly reduced NR activity by 29 or 35%, respectively, whereas at 6 h, the activity of the enzyme decreased by an average of 25%. At 21 h, Put increased NR activity by 63%, while Spd and Spm elevated the enzyme activity by 114%. NR activity, that was reduced by 0.1 mM Spm at 3 and 6 h, returned almost to control values when c-PTIO (an NO scavenger) was used, confirming that NO was involved in the inhibition of NR activity. Nitric oxide was also mediating the PA-increase of the enzyme activity at longer incubation times, evidenced when the raise in NR activity produced by 0.1 mM Spm at the longest incubation time returned to the value of the control in the presence of cPTIO. Neither the protein expression nor the nitrate content were modified by PAs treatments. The involvement of PAs and NO in the regulation of NR activity is discussed.

Nitric oxide inhibits nitrate reductase activity in wheat leaves

Nitrate reductase (NR), a committed enzyme in nitrate assimilation, is involved in the generation of nitric oxide (NO) in plants. In wheat leaf segments exposed to sodium nitroprusside (SNP) or S-nitrosoglutathione (GSNO), NR activity was significantly reduced to different degrees between 3 and 21h, whereas its activity was partially recovered when the NO scavenger cPTIO was used. At 21h, NR activity decreased from 38% with 10μM SNP to 91% with 500μM SNP, respect to the C values. S-nitrosoglutathione reduced NR activity between 18% and 26% only at 3h. When added directly to the incubation solution, NR activity was quickly and strongly inhibited more than 90% by 10 or 50μM SNP, whereas 10μM GSNO reduced the enzyme activity an average of 50%, at 30min of incubation. l-NAME and d-arginine (nitric oxide synthase (NOS) inhibitors) increased NR activity by 14% and 52% respectively, at 21h of exposure, leading us to suppose that endogenous NOS-dependent NO formation could also be modulating NR activity. NR protein expression was not affected by 10 or 100μM SNP at 3 or 21h of incubation, whereas nitration of tyrosines was not detected in the NR protein. Nitrates, which content increased along the time in the tissues, could be exerting a role in this regulation.

Comparative effectiveness of cattle manure, poultry manure, phosphocompost and fertilizer-NPK on three cropping systems in vertisols of semi-arid tropics. II. Dry matter yield, nodulation, chlorophyll content and enzyme activity

A field experiment was conducted on a deep Vertisol of Bhopal, India to compare root and shoot biomass, chlorophyll content, enzyme activity and nodulation in three cropping systems at three combinations of organic manure and inorganic-fertilizer: 75%NPK + 5 t farmyard manure (FYM), 75%NPK + 1.5 t poultry manure (PM), and 75%NPK + 5 t phosphocompost (PC) vis-a-vis 0%, 75% and 100% of fertilizer-NPK. In general, nodule number and its mass were lower in intercrop soybean than sole soybean. Also there was decrease in the nodule number with higher NPK dose. The FYM treated plots recorded 22.0% and 7.6% higher nodule mass than poultry manure and phosphocompost plots, respectively. Also, the total chlorophyll content was higher in organically treated plots than that in 100% NPK particularly at 30 days after sowing (DAS, pre-flowering). In sorghum the peak nitrate reductase (NR) activity was recorded at 60 DAS while in soybean it was at 30 DAS. The NR activity was higher in intercrop sorghum than that in sole sorghum. Maximum NR activity was observed in 100% NPK. Soybean/sorghum intercropping system recorded significantly higher root and shoot biomass than sole soybean and sorghum. The crop growth rates were relatively rapid during 30–60 DAS and followed the order; intercropping > sole sorghum > sole soybean. With the increase in NPK dose from 0% to 100% there was significant improvement in the dry matter (DM) production in sole sorghum and soybean/sorghum intercropping system. Soybean as preceding crop recorded the highest DM, chlorophyll content, NR activity in wheat while these values were the lowest in sorghum–wheat system.

Comparative studies of diurnal variations of nitrate reductase activity in wheat, oat and barley

Diurnal variations of in vitro and in vivo (intact tissue assay) nitrate reductase (EC 1.6.6.1) activity and stability were examined in leaves of wheat (Triticum aestivum L. cv. Runar), oat (Avcna saliva L. cv. Mustang) and barley (Hordeum vulgure L. cv. Agneta and cv. Gunillu). Nitrate reductase activity was generally higher for wheat than for oat and barley. However, the diurnal variations of nitrate reductase activity and stability were principally the same for all species, e.g. the high activity during the photoperiod was associated with low stability. All species showed a rapid (30‐60 min) increase in the in vitro and in vivo activity when the light was switched on. When light was switched off the in vitro activity decreased rapidly whereas decrease in in vivo activity was slower. These experiments support the hypothesis that an activation/ deactivation mechanism is involved in the regulation of diurnal variations in nitrate reductase activity. Red light enhanced nitrate reductase activity in etiolated wheat and barley leaves. In green leaves, however, the daily increase in nitrate reductase activity was not induced by a brief red light treatment. Indications of different regulation mechanisms for the diurnal variations of nitrate reductase activity among the cereals were not found.

Reevaluation of in vivo assay of nitrate reductase activity in wheat leaves

During in vivo assay of nitrate reductase in wheat leaf discs, the amount of nitrate released in the medium was greatly influenced by the pH of the medium, although it had no effect on total nitrite formed in the leaf tissue. In an acidic medium nitrite excreted out was negligible, while it increased considerably at alkaline pH-values. Extraction of leaf discs in boiling water resulted in significant loss of nitrite at acidic pH. In order to study the effect of malonate, succinate and L-malate, these acids were infiltrated into leaf discs at acidic pH while subsequent incubation and extraction was done at pH 7.5. It was observed that malonate and succinate significantly inhibited in vivo nitrate reduction while L-malate was without any effect.

Nitrate Uptake and the Induction of Nitrate Reductase Activity in Wheat (Triticum aestivumL.) Seedlings

Nitrate uptake and the subsequent induction of in vivo nitrate reductase activity in wheat were studied by investigating a euploid and certain ditelosomic stocks which exhibited in vivo activity significantly greater than that of the euploid. The kinetics of nitrate uptake were investigated, but the high activities of the ditelosomics were not caused by increased uptake of nitrate, although ditelo-7BL exhibited unusual uptake dynamics. Analysis of the induction of nitrate reductase activity revealed a biphasic general pattern, with an initial rapid phase being followed by a slower but longer period of induction. The induction rate over the second period, although responsible for only a minor proportion of the total activity induced, was positively correlated with the final nitrate reductase level, unlike the rate over the first induction period. Several stocks exhibited high induction rates over one or other of the two phases, while ditelo-lAs showed an abnormal monophasic induction pattern. At the end of the second period of induction, nitrate reductase activity became more or less steady, except for activity fluctuations associated with the time of application of induction stimuli.