Nitrate Reductase Enzyme Research Articles

Cumulative effect of gibberellic acid and phosphorus on crop productivity, biochemical activities and trigonelline production in Trigonella foenum-graecum L.

AbstractFenugreek (Trigonella foenum-graecum L.) is an antidiabetic plant. Its bioactive compound, trigonelline, is known to counter diabetes through insulin secretion, modulation of β cell regeneration and quick activity of glucose metabolism related enzymes. A pot experiment was conducted in the natural conditions of net house of the Department of Botany, Aligarh Muslim University, Aligarh (UP), India, to evaluate the effect of four concentrations of GA3 (0, 10−7 M, 10−6 M and 10−5 M), alone and in combination with phosphorus (40 kg P ha−1), on growth, biochemical and yield attributes of fenugreek. Compared to control, the combination of GA3 and phosphorus (P40 + 10−6 M GA3) significantly increased the activities of nitrate reductase (30.8%) and carbonic anhydrase (30.7%) enzymes; it also enhanced the seed yield (140.6%) and the content of total chlorophyll (28.5%) and carotenoids (26%). There was also significant increase (19.51%) in the content of seed trigonelline.

Over-expression of NtHb1 encoding a non-symbiotic class 1 hemoglobin of tobacco enhances a tolerance to cadmium by decreasing NO (nitric oxide) and Cd levels in Nicotiana tabacum

Tobacco gene NtHb1 encoding a non-symbiotic class 1 hemoglobin was identified to confer cadmium (Cd) tolerance in yeast Saccharomyces cerevisiae by expressing cDNA library of Nicotiana tabacum in WT yeast Y800. To examine the role of NtHb1 in plants, NtHb1 was over-expressed in tobacco (N. tabacum). NtHb1-expressing tobacco exhibited higher tolerance to Cd, but lower levels of Cd compared with vector-expressing control tobacco. Since it was reported that Hb (hemoglobin) is involved in scavenging NO, we investigated NO level in control and NtHb1-expressing tobacco. Interestingly, NO content of NtHb1-tobacco was lower than that of control tobacco in the presence of Cd, while there is no significant difference in NO accumulations of two plants in the absence of Cd. Among Cd transporters, CAX3 (cation/proton exchanger) transcript level of NtHb1-tobacco in response to Cd was higher than control tobacco, but IRT1 (iron-responsive transporter) expression was not. In addition, knockoutArabidopsis of AtNR1 and AtNR2 encoding NO producing enzymes nitrate reductase 1 and nitrate reductase 2 respectively, displayed enhanced Cd tolerance and reduced NO amounts in comparison with control Arabidopsis Col-0. Taken together, it is concluded that the increase of Hb by NtHb1 expression promotes Cd tolerance and reduces Cd accumulation by alleviating the level of NO induced by Cd in transgenic tobacco.

Simple Method for Measuring Endothelial Nitric Oxide Synthase Activity in Clinical Researches

Antiatherogenic Effect of Almond Oil in Streptozotocin Induced Diabetic RatsMona Anwar, Wafaa Ghoneim Shousha, Hatem A. El-mezayen, Raafat A wadallah, Maha El-Wassef, Naglaa M. Nazif and Mona A. El-bana

Some Growth Promoting Essential Mineral Elements Alleviates The Salinity Effect on Nitrate Reductase and Hill Reaction Activities in Cotton (Gossypium hirsutum) cv. “CIM 496”

The effect of salinity was observed on the activity of nitrate reductase and hill reaction activity in the leaves of Cotton (Gossypium hirsutum L.) cv. “CIM 496” grown in large size plastic pot culture, irrigated with sea salt concentrations of 0.0%(EC iw: 0.6 dS/m), 0.4%(EC iw: 6.2 dS/m) and 0.8%(EC iw: 10.8 dS/m) and subjected to foliar application with non-spray (control), water spray, KNO3 (500 ppm), KCl (500 ppm), Urea(1000 ppm), Fe-EDTA (5ppm), MnCl2 (5ppm) and MoO3 (5ppm) alone and their mixture. Foliar spray of the mixture showed better result as compared to that of above mentioned individual mineral. Conclusion made on the comparative performance of nitrate reductase and hill reaction enzymes under above number of treatments in the leaves of cotton growing under various sea salt concentrations is given below: Nitrate Reductase Activity (NRA): Nonspray< water spray< KCl< Mo< KNO3< Urea< KNO3 + Mo< KNO3 +Fe + Mn + Mo; Hill Reaction Activity (HRA): Nonspray< water spray< KNO3<Fe<Mn< KNO3 + Fe< KNO3 +Mn <KNO3 +Fe + Mn + Mo.

Effect of Pesticide on Nitrate Reductase Activity in Trigonella Foenum i.e., Fenugreek

Nitrate reductase (NR) is the key enzyme of nitrate assimilation subjected to inhibition by excessive use of pesticides. Now a day’s pesticides are used commonly at a large scale on field. In the present study, the effect of pesticides on nitrate reductase activity has been studied by in vivo assay in Trigonella foenum-graecum. The four pesticides Confidor, Omite, Karathane, Chlorpyrifos+Cypermethrinwere used for study at various concentrations such as 25, 50,100 and 200 μl/100 ml. The application of pesticides showed decrease in activity of nitrate reductase enzyme. Out of these pesticides confidor affected the nitrate reductase activity maximum. The nitrate reductase activity in control plant was found to be 0.460U ± 0.005 which was reduced up to 0.192 ± 0.008U in presence of confidor. Even in the tissue specific assay it was seen that the leaves and stem of the plant, the NR activity was affected maximum than other par

Physiological and biochemical changes in sugarcane in relation to anthocyanin accumulation during winter season

In order to evaluate ten sugarcane genotypes namely-CoS 07250, CoS 07240, CoS 08272, CoS 08279, CoS 09240, CoS 10239, CoSe 01434, CoSe 06455, CoSe 06456 and UP 05125 in relation to Anthocyanin accumulation, pot experiment was conducted in autumn season of 2011–2015, at the research farm of u.p Council of sugarcane research, Shahjahanpur (UP) India. The result of pooled data indicated that autumn planted young sugar plants accumulated anthocyanin pigment during winter season, which might have protected their photo system from adverse effects due to low temperature. Activity of nitrate reductase enzyme was higher during pre winter (November) which declined during the winter season (January). However, varieties CoS 07250, CoS 08279 and CoSe 01434 maintained higher activity of this enzyme throughout the formative phase of the crop. Total chlorophyll content in leaf followed the similar trend, being more during pre winter and declined in winter season with the increase of anthocyanin pigment. Crop growth rate (CGR) and Plastocrone were also higher in these varieties during the onset of summer. Thus greater accumulation of anthocyanin pigment in CoS 07250, CoS 08279 and CoSe 01434 during winter season resulted higher rate of metabolic activities to cope with low temperature stress.

Impact of Different Land use Management on Soil Enzyme Activities and Bacterial Genetic Finger Prints of North-Western Himalayas

Land uses has significant impact on soil biological properties that incessantly intimates the soil quality change and are assessed by soil microbial and biochemical indicators, as they are highly sensitive to change in environment. The purpose of this study was to investigate the impact of land use on soil enzyme activities and gene diversity in selected location of Northwestern Himalayas, India. Nine different land use system of similar soil type at depth 0-15cm were analyzed for soil enzymes (Dehydrogenase, Acid Phosphatase, Alkaline Phosphatase, Nitrate Reductase, Arylsulphatase, and Phytase) and genetic fingerprints (Randomly Amplified Polymorphic DNA) analysis. The land use systems investigated are Oak (Quercus incana), Deodar (Cedrus deodara), Pine (Pinus roxburghii) trees, Apple orchids and crop based systems in uplands and valleys. All the soil enzymes were significantly higher in forest ecosystem followed by organic farm and conventional maize-wheat farm soil. The principal component analysis (PCA) of nine different land use systems based on soil enzymes shows significant variation in data and all the long-term agricultural lands were segregated together. However maize-wheat and organic farm are group together in the PCA plot. Hierarchical clustering by wards method of soil enzymes clusters the deodar forest soil, oak forest soil and organic farming in one cluster and segregates remaining land use system in another. RAPD analysis showed high polymorphism between samples and similarity indexing using unweighted pair-group method with arithmetic averages resulted in four clusters. Land use showed significantly negative impact on soil enzymes and genetic fingerprints in long-term agricultural lands as compared to natural forest ecosystem and organic farming as reveal by RAPD assisted marker.

Cytokinin enhanced biomass and yield in wheat by improving N-metabolism under water limited environment

Drought is an important limiting factor for wheat production in most agricultural areas of the world. A study was conducted to analyze the effect of cytokinin [benzyl amino purine (BAP), 40 µM] on growth and yield vis-a-vis nitrogen metabolism in two contrasting wheat cultivars, viz., C-306 (drought tolerant) and PBW-343 (drought susceptible) under water deficit stress condition. Water deficit stress significantly decreased yield and total biomass, while cytokinin-treated wheat plants retained higher biomass and yield. Positive effect of cytokinin on yield and biomass was due to cytokinin induced N-metabolism under both water regimes. Water deficit stress decreases activity of two major enzymes of nitrogen assimilation pathway, viz, nitrate reductase (NR) and glutamine synthetase (GS) and increased total protease activity. Cytokinin enhanced the activity of NR and GS enzymes and reduced the protease activity in both the cultivars and water regimes, which also led to increase in level of total nitrogen and total protein content. Overall, cytokinin was able to delay senescence of leaves under water deficit stress conditions by enhancing N-metabolism in general and specifically at the reproductive stage, contributing to increase in grain yield of both the cultivars under both the water regimes.

Plasmonic photocatalysis and kinetics of reactive dye degradation in aqueous solution using enzymatically synthesized Ag/ZnO

Nano-composite of Ag/ZnO was prepared in the presence of Escherichia coli nitrate reductase enzyme with silver from AgNO3 over ZnO. Ag/ZnO produced was characterized using field emission scanning electron microscope, energy-dispersive spectroscopy, transmission electron microscope, X-ray diffraction, porosimetry system and UV–Vis–NIR spectrophotometer. Prepared catalyst was used to oxidize Reactive Black B in aqueous solution under visible light. Effects of silver concentration, initial dye concentration, pH, catalyst dosage and reusability of the composites were studied. The photo degradation followed pseudo-first order kinetics. Further, the potential of catalyst for decolourisation of textile industry effluent were also examined and it was observed that 40 % COD reduction and 100 % colour removal was achieved within 120 min of reaction.

In vivo synthesis of selenium nanoparticles by Halococcus salifodinae BK18 and their anti-proliferative properties against HeLa cell line.

Nanoparticles synthesis by bacteria and yeasts has been widely reported, however, synthesis using halophilic archaea is still in a nascent stage. This study aimed at the intracellular synthesis of selenium nanoparticles (SeNPs) by the haloarchaeon Halococcus salifodinae BK18 when grown in the presence of sodium selenite. Crystallographic characterization of SeNPs by X-ray diffraction, Selected area electron diffraction, and transmission electron microscopy exhibited rod shaped nanoparticles with hexagonal crystal lattice, a crystallite domain size of 28 nm and an aspect ratio (length:diameter) of 13:1. Energy disruptive analysis of X-ray analysis confirmed the presence of selenium in the nano-preparation. The nitrate reductase enzyme assay and the inhibitor studies indicated the involvement of NADH-dependent nitrate reductase in SeNPs synthesis and metal tolerance. The SeNPs exhibited good anti-proliferative properties against HeLa cell lines while being non-cytotoxic to normal cell line model HaCat, suggesting the use of these SeNPs as cancer chemotherapeutic agent. This is the first study on selenium nanoparticles synthesis by haloarchaea.

BIOTECHNOLOGY STUDY TO REMOVE NITRATE AND NATRATE BY USING PSEUDOMONAS AERUGINOSA STRAINS ISOLATED FROM WEST WATER IN BAGHDAD CITY

This isolated genus has a considerable and important value in biotechnological resources and applications,. These bacteria use much more efficiently the organic substances small molecules units such nitrate, nitrite and used in biosorption for heavy metals ions like Zn+2,Cu+2, Pb+2. Cd+2 This work belongs to a research study on biodegradation of environmental pollutants with nitrate and nitrite, which studies denitrifying bacteria such Pseudomonas aeruginosa. They were tested in labs and naturally in waste water of deferent areas at Baghdad.It was found that strains of Pseudomonas were reduced of 21-35 % from NO3- and 31-47% from NO2- during 24 hours incubation ,checking pH7, contained in used medium to some other nitrogenous compounds using the enzyme nitrate reductase and enzyme nitrite reductase. In the process of bio sorption showed the removal that the concentration of heavy metals( Zn,Cu,Cd,Pb) above from the type of bacteria. The ratio of removal for Pseudomonas aeruginosa in concentration above 48.4% ، 85.6% ،82.3% ،33%.

Anti-bacterial activities of silver nanoparticles synthesized from plant leaf extract of Abutilon indicum (L.) Sweet

Abutilon indicum (L.) Sweet is an Asian phytomedicinal plant and it has hypoglycemic and anti-bacterial activities. The nano silver was successfully synthesized from leaf extract using A. indicum (L.) Sweet. Nitrate reductase enzyme or other extra cellular proteins released from leaf extract involve in reduction of nitrate and formation of silver ions occurs. These proteins or enzymes act as template for the silver nucleation sites for the formation of silver nanocrystals and also act as capping agents (peptides or proteins) which control the size and shape of silver nanoparticles. FT-IR reveals that protein molecules can be bound to silver nanoparticles and XRD result confirmed crystal structure of nanoparticles. On the basis of our results, extract of A. indicum (L.) Sweet leaf protein molecules plays a vital role in reducing and capping of silver nanoparticles at 50 μl concentrations which act as anti-bacterial agent to control pathogenic microorganisms of Klebsiella pneumonia, Salmonella typhi, Bacillus subtilis, and Proteus vulgaris.

Green Synthesis of Silver Nanoparticles by Haloarchaeon &lt;i&gt;Halococcus salifodinae &lt;/i&gt;BK6

Nanobiotechnology is a multidisciplinary branch of nanotechnology which includes fabrication of nanosized materials using biological approaches. Highly structured metallic and metal sulfide nanoparticles have been reported to be synthesized by numerous bacteria, fungi, yeasts and viruses. However, biosynthesis of nanoparticles by Haloarchaea (salt-loving archaea) of the third domain of life, Archaea, is in its nascent stages. In this study, we report the intracellular synthesis of stable, mostly spherical silver nanoparticles (SNPs) by the haloarchaeal isolateHalococcus salifodinaeBK6. The isolate adapted to silver nitrate was found to exhibit growth kinetics similar to that of cells unexposed to silver nitrate. The nitrate reductase enzyme assay and the enzyme inhibitor studies showed the involvement of NADH dependent nitrate reductase in silver tolerance, reduction, and synthesis of SNPs. UV visible spectroscopy, XRD, TEM and EDAX were used for characterization of SNPs. The XRD exhibited characteristic Bragg peaks of face centered cubic silver with crystallite domain size of 26 nm and 12 nm for SNPs synthesized in NTYE and halophilic nitrate broth, respectively. TEM analysis exhibited an average particle size of 50.3 nm and 12 nm for SNPs synthesized in NTYE and halophilic nitrate broth (HNB), respectively. The as synthesized SNPs exhibited antimicrobial activity against both Gram positive and Gram negative organisms.

English

Chickpea (Cicer arietinum L.) is one of the oldest pulse crops cultivated from ancient time both in Asia and Europe. India is the largest producer of chickpea in the world, sharing 65 and 70% of production and total global area, respectively. The inhibition of plant growth and crop productivity by excess heavy metals is a global agricultural problem. Therefore, a pot experiment was conducted at wire house of the Department of Plant Physiology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi during Rabi season of 2009-10, using a complete randomized design with four replications and five treatments. Our experimental findings reveal that excessive amount of copper (Cu) (200 mg/kg) and arsenic (As) (200 mg/kg) in soil caused a mark reduction in nitrate reductase enzyme activity at both growth stages. However, a little increment was found in proline content, peroxidase and superoxide dismutase activity under higher doses of Cu and As at both growth stages. The study aimed to get more information on physiological changes in activity of antioxidant enzymes under heavy metal toxicity. Key words: Antioxidant enzymes, chickpea, heavy metals, nitrate reductase, proline content.

Pigments apparatus and anthocyanins reactions of borage to irrigation, methylalchol and titanium dioxide

This study was done to evaluate the effect of methanol and nano TiO2 foliar application on photosynthetic apparatus, anthocyanin and changes of nitrate reductase in borage (Borago officinalis L.) under deficit irrigation. Tests were done as a split factorial experiment set up as a completely randomized block design with four replications, in the year 2012, at Shahriyar City, Iran. Irrigation conditions were the factors tested in the main plots: deficit irrigation condition (irrigation every 7 days) and irrigation every 14 days. Methanol solutions 0 (control or sprayed with water), 15%, 35% and 45% (v/v) and, nanoTiO2 treatment (0 or control and nano TiO2 at concentrations of 0.01%, 0.03% and 0.05%) were tested in the sub plots. Plant characteristics under evaluation were those of; chlorophyll (Chl) a,b and total chlorophyll, Spad value, net photosynthetic rate, rubisco carboxylase activity, anthocyanin and nitrate reductase enzyme (NR). Also evaluated were the effects of interaction of deficit irrigation and methanol on all measured traits, and results determined the effect as significant with the exception of NR and interactions of deficit irrigation and nano TiO2 on all measured traits. Test results showed that maximum amount of traits at deficit irrigation was achieved by spraying with 45% v/v oncentration of methanol and nano Tio2 at the concentration of 0.05%. But the best treatments under normal irrigation were 15% v/v of methanol and 0.03% of nano Tio2. but at the methanol concentration of 45% and non-application of nano Tio2 under normal irrigation had the least effect. In summary, results of these tests determined that application of methanol and nano Tio2, can promote plant resistance to drought stress (particularly borage plants).

Remediation of Nickel-Contaminated Soil by Brassica juncea L. cv. T-59 and Effect of the Metal on Some Metabolic Aspects of the Plant

ABSTRACTAmong four cultivars of Brassica juncea L., viz., TM-4, TM-2, RH-30, and T-59, cv. T-59 was relatively more tolerant to nickel (Ni) toxicity based on the growth parameters, seedling vigor index, and metal tolerance index. Nickel application inhibited the activity of the nitrate-assimilating enzyme nitrate reductase in the roots, stem, and leaves, whereas the total organic nitrogen, proline, and activity of a polyamine-metabolizing enzyme, diamine oxidase, increased in this tolerant cultivar (T-59). It accumulated a good amount of Ni from the soil in its root and shoot (i.e., 6.0–6.51 μg Ni g−1 dry weight) during 2 months of cultivation with an 8.0 mM Ni supply in the soil. The data presented in this paper indicate that Ni tolerance and its removal by Indian mustard from subtropical Indian soil is cultivar dependent, possibly due to different genetic and physiological adaptations of the cultivars.

A Lotus japonicus mutant defective in nitrate uptake is also affected in the nitrate response to nodulation

A chlorate-resistant mutant (Ljclo1) of the model legume Lotus japonicus was identified that showed normal levels of nitrate reductase enzyme activity but had decreased uptake of nitrate, as determined from nitrate depletion and electrophysiological measurements. The data suggest that the mutant could be affected specifically in the low-affinity but not in the high-affinity nitrate transport system, and also showed decreased uptake of chlorate. Back-crosses of the mutant plant to the wild type indicated that it is affected in a single Mendelian recessive trait. Thus, the mutation produced in Ljclo1 may be related to some of the low-affinity nitrate transporters or to a regulatory mechanism associated with nitrate/chlorate uptake. Both size and chlorophyll content in young leaves of the mutant plants were significantly reduced compared to the wild type. In addition, nodulation performance of the mutant plants was similar to the wild type in the absence of any exogenous nitrate. However, the nodule:root biomass ratio in mutant plants was considerably reduced in the presence of 1-2mm nitrate. The levels of several transcripts for nitrate transport and assimilation genes were determined for the wild type and mutant plants and were slightly different. The results suggest interdependence between nitrate uptake, plant growth and nodulation in Ljclo1 mutant plants.

Potassium Nitrate Priming Affects the Activity of Nitrate Reductase and Antioxidant Enzymes in Tomato Germination

Priming has been used to improve the performance of germination at the field, and potassium nitrate (KNO3) is a promising compound for this purpose. The nitrate (NO3) could be absorbed, being used in the metabolism of the embryo, through the enzyme nitrate reductase (NR). Besides, the priming could also activate the response of the antioxidant system, becoming the primed seeds more prepared for possible stresses. Thus, the aim of this study was to evaluate the metabolic effect of nitrate in tomato seed germination by the quantification of NR activity, and also evaluate the activity of some antioxidant enzymes as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX). Tomato seeds were primed using solutions of polyethylene glycol (PEG 6000) -1,1MPa, 50 mM KNO3 and PEG+KNO3. The variables analyzed were germination (germinability, mean germination time, mean germination rate, coefficient of variation of the germination time, uncertainty and synchrony) nitrogen, total proteins and enzymes. The germination data were analyzed using an ANOVA, comparing the averages by Scott-Knott test (P < 0.05). To analyze the nitrogen, protein and enzymatic activities, we used a Kruskal-Wallis test (P < 0.05). The results show an increase in the NR activity, as well as in the antioxidant enzymes. The germination time (t) and germination rate (v) primed in KNO3 had a better performance compared to the other treatments. In conclusion, the observed benefits in tomato seeds primed with KNO3 were related to the activity of the enzyme nitrate reductase in the production of nitrite/nitric oxide, which acted promoting a faster germination.

Effect of vermicompost tea on yield and nitrate reductase enzyme activity in saladette tomato

In conventional agriculture, heavy doses of chemical fertilizers and pesticides are often used to improve the yield of various horticultural crops. These chemical fertilizers and pesticides cause health problems among consumers. Due to the adverse effects of chemical fertilizers, there is interest in the use of organic fertilizers. It has long been recognized that the rate-limiting step for nitrogen assimilation, the reduction of NO 3 to NO 2 catalyzed by nitrate reductase (NR), is highly regulated. An increase in the amount and activity of NR leads to a corresponding increase in the potential for nitrate reduction and confers a greater capacity for general amino-acid synthesis, protein synthesis or total nitrogen assimilation. The aim of the current experiment was to determine the effect of organic and conventional fertilizers on yield and nitrate reductase activity in saladette tomato.Tomato plants were grown under controlled greenhouse conditions and treated with either organic or conventional fertilizer. We evaluated five treatments: F1, sand + inorganic fertilizer; F2, sand + vermicompost tea; F3, a 1:1 mixture of sand: compost + vermicompost tea; F4, a 1:1 mixture of sand: vermicompost + vermicompost tea; and F5, a 2:1:1 mixture of sand: compost: vermicompost + vermicompost tea. The evaluated variables were yield, fruit size, number of fruits, fruit quality, chlorophyll content, and in vivo nitrate reductase enzyme activity. Fertilizer type strongly influenced the yield, fruit size, and fruit quality. The best organic fertilizer for tomatoes was sand + vermicompost tea. Tomatoes in this treatment group produced the second highest yield, the best NO 3 assimilation, the greatest nitrate reductase

Biosynthesis of Silver Chloride Nanoparticles Using Bacillus subtilis MTCC 3053 and Assessment of Its Antifungal Activity

The present investigation reported the synthesis of silver chloride nanoparticles using Bacillus subtilis. The adsorption of colloidal silver chloride nanoparticles showed an intense peak at the wavelength of 400 nm after 20 hrs of biomass incubation. The size of the silver nanoparticles ranges from 20 to 60 nm which was obtained from transmission electron microscope (TEM). The X-ray diffraction (XRD) pattern confirmed the crystalline nature of the nanoparticles. The bright circular spots of selected diffraction area (SAED) pattern also confirmed the good crystalline nature of the silver chloride nanoparticles with high magnification of TEM images. The presence of nitrate reductase enzyme in the cellular membrane of B. subtilis was confirmed by sodium dodecyl (SDS) polyacrylamide gel electrophoresis and it was found that the molecular weight is 37 kDa. The possible functional groups of the reductase enzyme in B. subtilis were identified by Fourier transform infrared spectroscopy (FTIR). Finally, antifungal activity of silver chloride nanoparticle was examined against Candida albicans, Aspergillus niger, and Aspergillus flavus. We conclude that the synthesis of silver chloride nanoparticles using microorganisms is more economical and simple. The antifungal property of silver chloride nanoparticles will play a beneficial role in biomedical nanotechnology.