Induction Of Nitrate Reductase Research Articles

Mechanisms of chlorate toxicity and resistance in Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an opportunistic bacterial pathogen that often encounters hypoxic/anoxic environments within the host, which increases its tolerance to many conventional antibiotics. Toward identifying novel treatments, we explored the therapeutic potential of chlorate, a pro-drug that kills hypoxic/anoxic, antibiotic-tolerant P. aeruginosa populations. While chlorate itself is relatively nontoxic, it is enzymatically reduced to the toxic oxidizing agent, chlorite, by hypoxically induced nitrate reductase. To better assess chlorate's therapeutic potential, we investigated mechanisms of chlorate toxicity and resistance in P. aeruginosa. We used transposon mutagenesis to identify genes that alter P. aeruginosa fitness during chlorate treatment, finding that methionine sulfoxide reductases (Msr), which repair oxidized methionine residues, support survival during chlorate stress. Chlorate treatment leads to proteome-wide methionine oxidation, which is exacerbated in a ∆msrA∆msrB strain. In response to chlorate, P. aeruginosa upregulates proteins involved in a wide range of functions, including metabolism, DNA replication/repair, protein repair, transcription, and translation, and these newly synthesized proteins are particularly vulnerable to methionine oxidation. The addition of exogenous methionine partially rescues P. aeruginosa survival during chlorate treatment, suggesting that widespread methionine oxidation contributes to death. Finally, we found that mutations that decrease nitrate reductase activity are a common mechanism of chlorate resistance.

Inhibition of N2 Fixation by N Fertilization of Common Bean (Phaseolus vulgaris L.) Plants Grown on Fields of Farmers in the Eastern Cape of South Africa, Measured Using 15N Natural Abundance and Tissue Ureide Analysis

Inhibition of N2 fixation in N-fertilized common bean (Phaseolus vulgaris L.) plants growing on the fields of farmers in the Eastern Cape of South Africa was measured using 15N natural abundance and tissue ureide analysis. The N-fertilized bean plants revealed greater soil N uptake, higher concentrations of nitrate in organs, low tissue ureide levels, and much lower percent relative ureide-N abundance when compared with unfertilized plants. In contrast, the unfertilized plants showed greater nodule fresh weight, higher N derived from fixation (e.g., 84.6, 90.4, and 97.1% at Lujecweni fields 2, 3, and 4, respectively), increased amount of N-fixed (e.g., 163.3, 161.3, and 140.3 kg ha−1 at Lujecweni fields 2, 3, and 4, respectively), greater ureide concentration in stems and petioles, higher % relative ureide-N abundance, and low soil N uptake. We also found that the percent N derived from fixation (%Ndfa) was very high for some bean plants receiving a double dose of N fertilizer [e.g., Lujecweni field 1 (51.8%) and Tikitiki field 1 (53.3%], and quite high for others receiving a single dose of N fertilizer [e.g., Tikitiki field 2 (50.1%), Mfabantu fields 1 and 2 (45.5 and 79.9%, respectively), and St. Luthberts field 1 (58.9%)]. Though not assessed in this study, it is likely that the rhizobia that effectively nodulated the N-fertilized bean plants and fixed considerable amounts of symbiotic N had constitutive and/or inducible nitrate reductase genes for reducing nitrate in nodules and bacteroids, hence their ability to form root nodules and derived high %Ndfa in bean with added N. While single- and double-dose N fertilizer applications increased plant growth and grain yield compared to unfertilized bean plants, the single-dose N fertilizer application produced much greater grain yield than the double dose. This indicates that farmers should stop using a double dose of N fertilizers on bean production, as it decreases yields and can potentially pollute the environment. This study has however shown that government supply of free N fertilizers to resource-poor farmers in South Africa increased bean yields for food/nutritional security.

Salt tolerance of nitrate reductase in Halomonas sp. B01

A systematic study on the lack of dissimilatory nitrate reductase (NAR) properties in Halomonas strains had been reported so far. The effects of different factors on Halomonas sp. B01 NAR activity were investigated. The salt tolerance of NAR was characterized. The denitrification process under high salt conditions was reported. Halomonas sp. B01 expressed membrane-bound NAR under induced culture by nitrate. The optimum pH of the enzyme reaction system was 8, and the optimum temperature was 30°C. The mRNA expression abundance of narH in NAR encoding gene was highest in the 60g/L NaCl inducing matrix. The NaCl concentration of optimum growth and induction of NAR were both 60g/L. The ectoine added to the NAR vitro enzyme reaction system could maintain NAR activity under high NaCl concentration. In the range of 0-60g/L NaCl, the NAR activity was stable at 17.7 (± 0.3) U/mg. The denitrification was performed by Halomonas sp. B01 at 60g/L NaCl, and the denitrification rate reached 97.1% at 24h. This study reveals for the first time the NAR properties of Halomonas strains, which provides a theoretical and technical basis for the nitrogen removal of high-salt nitrogenous wastewater using this strain.

Nitrate reductase activities in plants from different ecological and taxonomic groups grown in Japan

AbstractPlants generally use soil inorganic nitrogen, ammonium (NH4+–N), and nitrate (NO3−–N) as sources of nitrogen, an essential nutrient. The assimilation processes after uptake differ considerably from ammonium to nitrate. Nitrate must be reduced to ammonium in plant tissue before it is synthesized to amino acids, while ammonium is directly and immediately synthesized to amino acids after its uptake. Nitrate reductase is an enzyme that catalyzes the first and rate‐limiting step of nitrate assimilation, reducing nitrate to nitrite. It is a substrate‐inducible enzyme, and the capacity to induce nitrate reductase varies greatly among plant species. In vivo nitrate reductase activity (NRA) is generally measured as a nitrite production rate during incubation using fine cut pieces of plant tissue, and it is applicable as an indicator on plant nitrate use. Here we present in vivo NRA of leaves from a total of 108 species including arboreal trees, small trees, shrubs, herbs, a vine and a moss. For 75 of the species, NRA in fine roots was also determined. At least 20 species in sampled plants were imported and planted for scientific or industrial purposes, but most sampled species were native to Japan. Several inventory studies of plant NRA have been conducted, mainly in Europe, and they provided information on the species‐specific capacity of nitrate use by plants in Europe. However, to our best knowledge, there has been hitherto no published inventory of the NRA of plants in Japan where many endemic species are distributed. Our dataset contains plant NRA with species, family name, life form, leaf lifespan (evergreen or deciduous), growth stage, the season of sample collection, growth conditions (natural or cultivated) and other treatments/conditions when applicable. The data provided by this study may contribute to future works that require information regarding the plant species characteristics for nitrate use capacity or nitrate preference.The complete data set for this abstract published in the Data Paper section of the journal is available in electronic format in MetaCat in JaLTER at http://db.cger.nies.go.jp/JaLTER/metacat/metacat/ERDP-2019-06.1/jalter-en. [Correction added on 7 September 2020, after first online publication: JaLTER URL has been updated.]

Cold modulated nuclear S-nitrosoproteome analysis indicates redox modulation of novel Brassicaceae specific, myrosinase and napin in Brassica juncea

Stress leads to production of reactive oxygen species (ROS) and reactive nitrogen species (RNS). Among the RNS, nitric oxide (NO) is an important signaling molecule that mediates its effect majorly via a reversible post-translational modification (PTM) S-nitrosylation. However, information on the NO mediated cold stress signaling is still missing for nucleus. Therefore, in the present study an attempt was made to analyze NO signaling, particularly, S-nitrosylation in the nuclei of Brassica juncea seedlings. Cold stress (4 °C) induced nitrate reductase (NR) mediated NO production along with the accumulation of available protein thiols (5 fold, at 96 h, 4 °C) and low molecular weight thiols (1.2 fold at 12 h, 4 °C). S-nitrosoglutathione (GSNO, a NO donor) and cold (endogenous, S-nitrosylation without the addition of NO donor) treated, neutravidin-agarose affinity chromatography purified S-nitrosylated proteins were resolved as 55- and 32-spots respectively on the 2-D gels. Functional categorization after MALDI-TOF/TOF identification showed 67% targets to be stress/defense/redox related and 33% to be metabolic, suggesting the potential role(s) of S-nitrosylation in these responses. Cold stress increased S-nitrosylation of Cu/Zn SOD, l-ascorbate oxidase, fructose bisphosphate aldolase (FBA2) and malate dehydrogenase 1 (MDH1), therefore, indicating NO mediated regulation of redox and metabolism related pathways. Identification of targets of glucosinolate hydrolysis pathway (myrosinase and myrosinase-binding protein 2-like) indicated the role of S-nitrosylation in regulating mustard oil bomb. Presence of myrosinase was confirmed by activity measurements and western blotting. Increased S-nitrosylation of myrosinase in cold stress in the nucleus is reported for the first time. Here, we proposed S-nitrosylation mediated nuclear trafficking of non-nuclear targets such as beta-d-xylosidase, daikon cysteine protease, and beta-glucosidase to the nucleus. Similarly, identification of FBA2 and MDH1 in the nucleus, indicated the role of NO in modulating their moonlighting function via S-nitrosylation in stress. Present study provided the first repertoire of cold modulated S-nitrosylated proteins in the nucleus.

Ectopic Expression of PII Induces Stomatal Closure in Lotus japonicus.

The PII protein in plants has been associated to many different tissue specialized roles concerning the Nitrogen assimilation pathways. We report here the further characterization of L. japonicus transgenic lines overexpressing the PII protein encoded by the LjGLB1 gene that is strongly expressed in the guard cells of Lotus plants. Consistently with a putative role played by PII in that specific cellular context we have observed an alteration of the patterns of stomatal movement in the overexpressing plants. An increased stomatal closure is measured in epidermal peels from detached leaves of normally watered overexpressing plants when compared to wild type plants and this effect was by-passed by Abscisic Acid application. The biochemical characterization of the transgenic lines indicates an increased rate of the Nitric Oxide biosynthetic route, associated to an induced Nitrate Reductase activity. The phenotypic characterization is completed by measures of the photosynthetic potential in plants grown under greenhouse conditions, which reveal a higher stress index of the PII overexpressing plants.

Salinity-induced accumulation of endogenous H2S and NO is associated with modulation of the antioxidant and redox defense systems in Nicotiana tabacum L. cv. Havana

Salinity is one of the abiotic factors that most affect crop growth and production. This study focused on the effect of high salinity on the endogenous levels of the signaling molecules hydrogen sulfite (H2S) and nitric oxide (NO) in Nicotiana tabacum leaves and the extent of these for the biochemically-driven plant tolerance to such abiotic stress. The NaCl treatment for 10days led to an expressive augment of H2S and NO levels. This increase was correlated with the raise of l-Cys and l-Arg and the induction of l-cysteine desulfhydrase, cyanoalanine synthase, cysteine synthase, nitrate reductase and arginase, enzymes known to be involved in the biosynthesis of H2S or NO. The enzymatic antioxidant system (superoxide dismutase and catalase activity) was boosted and the non-enzymatic antioxidant glutathione was intensively oxidized in leaves upon stress allowing plants to cope with oxidative stress. Lower stomatal conductance was observed in stressed plants in comparison with control ones. Moreover, the high activity of antioxidant enzymes and high rate of glutathione oxidation following salt stress were considerably decreased upon NO or H2S scavenging. Thus, increment in NO and H2S levels and their interplay, along with metabolic and physiological changes, contributed to tobacco survival to extreme salinity conditions.

Controlled biosynthesis of silver nanoparticles using nitrate reductase enzyme induction of filamentous fungus and their antibacterial evaluation

The controlled synthesis of silver nanoparticles (AgNPs) using cell-free filtrate of Fusarium oxysporum fungus was investigated. The effect of fungal incubation period on nanoparticle formation and nitrate reductase enzyme activity was studied using UV–visible spectroscopy and Harley assay, respectively. The highest AgNP formation was observed in the cell-free filtrate of biomass harvested at the early stationary phase where the NR enzyme activity is the maximum. Mixing of the cell-free filtrates of fungal cultures obtained at 23, 28, and 33 °C with silver nitrate solution confirms the higher productivity of AgNP biosynthesis using the cell-free filtrate of fungus incubated at 28 °C. The effect of some factors such as carbon and nitrate sources and light in fungal incubation period on nitrate reductase induction and AgNP formation was also evaluated. In conclusion, increasing nitrate and carbon sources and presence of light induced NR enzyme and produced AgNPs with smaller size, higher monodispersity, and productivity. Results revealed that the presence of ammonium prevents the NR enzyme secretion and causes to the lower productivity of AgNPs.

Properties of nitrogen fertilization are decisive in determining the effects of elevated atmospheric CO2 on the activity of nitrate reductase in plants

ABSTRACTThe concentration of atmospheric CO2 is predicted to double by the end of this century. The response of higher plants to an increase in atmospheric CO2 often includes a change in nitrate reductase (NR) activity. In a recent study, we showed that, under elevated CO2 levels, NR induction in low-nitrate plants and NR inhibition in high-nitrate plants are regulated by nitric oxide (NO) generated via nitric oxide synthases. This finding provides an explanation for the diverse responses of plants to elevated CO2 levels, and suggests that the use of nitrogen fertilizers on soil will have a major influence on the nitrogen assimilation capacity of plants in response to CO2 elevation.

Nitrate reductase in detached embryos may serve as a marker of the preharvest tolerance of wheat seeds

We have observed an easily determinable parameter indicative of genetic pre-harvest sprouting (PHS) tolerance – the presence of an endosperm factor, presumably ABA, capable of inhibiting nitrate reductase (NR) induction in the embryo in the presence of NO3. This finding has importance not only for the early rapid screening of PHS tolerant of cereal cultivars but may also be an important tool to determine the mechanism of NR inhibition either by genetic repression or by post-translational down regulation of NR activity. In this work we sought a simple relationship between ABA content and NR activity level that we assumed to be closely related to PHS susceptibility. Studied the relationship between ABA content, dry weight and ability to germinate in seeds of examined wheat varieties resistant to preharvest sprouting Lutescens 70, and unstable – Novosibirskaya 67. The data obtained are shown that the level of ABA reaching its maximum to 40-45 days . The maximum content of ABA for both varieties accounted for 40 days after flowering, when the grains reach maximum wet weight in the future, as the ripening hormone levels quickly decreased with the decrease in fresh weight of grain . The level of ABA in embryos unstable to pre-harvest sprouting wheat Novosibirskaya 67 ranged from 20 to 40% below the level of the hormone in the grains resistant Lutescens 70 wheat. The results showed a significant increase in the ability to germinate, since it is a phase 40-45 DAP. Reduction of endogenous ABA content in grains at the final stage of maturation, with a relatively high percentage of germination, can be explained by the need and ensuring to the start of the germination process of the seed.

Three transcription regulators of the Nss family mediate the adaptive response induced by nitrate, nitric oxide or nitrous oxide in Wolinella succinogenes.

Sensing potential nitrogen-containing respiratory substrates such as nitrate, nitrite, hydroxylamine, nitric oxide (NO) or nitrous oxide (N2 O) in the environment and subsequent upregulation of corresponding catabolic enzymes is essential for many microbial cells. The molecular mechanisms of such adaptive responses are, however, highly diverse in different species. Here, induction of periplasmic nitrate reductase (Nap), cytochrome c nitrite reductase (Nrf) and cytochrome c N2 O reductase (cNos) was investigated in cells of the Epsilonproteobacterium Wolinella succinogenes grown either by fumarate, nitrate or N2 O respiration. Furthermore, fumarate respiration in the presence of various nitrogen compounds or NO-releasing chemicals was examined. Upregulation of each of the Nap, Nrf and cNos enzyme systems was found in response to the presence of nitrate, NO-releasers or N2 O, and the cells were shown to employ three transcription regulators of the Crp-Fnr superfamily (homologues of Campylobacter jejuni NssR), designated NssA, NssB and NssC, to mediate the upregulation of Nap, Nrf and cNos. Analysis of single nss mutants revealed that NssA controls production of the Nap and Nrf systems in fumarate-grown cells, while NssB was required to induce the Nap, Nrf and cNos systems specifically in response to NO-generators. NssC was indispensable for cNos production under any tested condition. The data indicate dedicated signal transduction routes responsive to nitrate, NO and N2 O and imply the presence of an N2 O-sensing mechanism.

The role of light in the inducation of nitrate reductase and nitrite reductase in cucumber seedlings

The activity of nitrate reductase (NR) and nitrite reductase (NiR) was investigated in vivo and in vitro in the roots and NR activity in 3-day-old cotyledons of cucumber seedlings. NR activity in the roots appears almost immediately after addition of nitrate ions to the induction medium, whereas, in the cotyledones NR induction is delayed. In general light enhances NR activity in the cotyledons and depresses it in the roots in experiments of short duration. Etiolation of the cotyledons reduces NR activity in the roots and leads to disappearance of the activity of this enzyme in the cotyledons, whereas the NR activity of roots kept in darkness, after transfer of the etiolated plants to light, increases threefold. In roots growing in darkness a delay in NiR induction is observed, while in those growing in ligth it occurs at the same time as NR induction. Chlormaphenicol (CAP), cycloheximide (CHI) and actinomycin D (ACM) applied at the beginning of the period of seedling induction with initrates inhibit NR activity in the cotyledons, whereas in the roots only CHI and ACM exert such an effect. To sum up, NR is synthesized in cucumber roots and cotyledons de novo on the cytoplasmic polyribosomes, and light per se is not indispensable for this synthesis, but it has an indirect influence on the activity level of NR and NiR both in the roots and the cotyledons.

Sulfate resupply accentuates protein synthesis in coordination with nitrogen metabolism in sulfur deprived Brassica napus

To investigate the regulatory interactions between S assimilation and N metabolism in Brassica napus, de novo synthesis of amino acids and proteins was quantified by 15N and 34S tracing, and the responses of transporter genes, assimilatory enzymes and metabolites pool involving in nitrate and sulfate metabolism were assessed under continuous sulfur supply, sulfur deprivation and sulfate resupply after 3 days of sulfur (S) deprivation. S-deprived plants were characterized by a strong induction of sulfate transporter genes, ATP sulfurylase (ATPS) and adenosine 5′-phosphosulfate reductase (APR), and by a repressed activity of nitrate reductase (NR) and glutamine synthetase (GS). Sulfate resupply to the S-deprived plants strongly increased cysteine, amino acids and proteins concentration. The increase in sulfate and cysteine concentration caused by sulfate resupply was not matched with the expression of sulfate transporters and the activity of ATPS and APR which were rapidly decreased by sulfate resupply. A strong induction of O-acetylserine(thiol)lyase (OASTL), NR and GS upon sulfate resupply was accompanied with the increase in cysteine, amino acids and proteins pool. Sulfate resupply resulted in a strong increase in de novo synthesis of amino acids and proteins, as evidenced by the increases in N and S incorporation into amino acids (1.8- and 2.4-fold increase) and proteins (2.2-and 6.3-fold increase) when compared to S-deprived plants. The results thus indicate that sulfate resupply followed by S-deprivation accelerates nitrate assimilation for protein synthesis.

Nitrate reductase and acid phosphatase activities as affected by inorganic phosphate in corn roots

The deficieny of inorganic phosphate in nutrient solution reduces by about 50 per cent NO<sub>3</sub>- absorption in corn seedlings, it decreases both <em>in vitro</em> and in vivo nitrate reductase (NR) activity, as well the potential and actual NR level and has a very weak effect on NR induction. Acid phosphatases activities increase in corn roots when the plants are grown in nutrient solution without phosphorus. We suggest that inorganic phosphate is required mainly for maintenance of NR activity rather, than for induction <em>in vivo</em> of nitrate reductase. It is not excluded that deficiency of inorganic phosphate in root tissue may be partly supplemented as the result of enhanced acid phosphatase activity.

Nitrate/oxygen co‐sensing by an NreA/NreB sensor complex of Staphylococcus carnosus

In Staphylococci maximal induction of nitrate reductase (narGHJI genes) requires anaerobic conditions, the presence of nitrate, and the NreABC regulatory system. Aerobic regulation is effected by the NreB/NreC two-component system. The role of the nitrate receptor NreA in nitrate induction and its relation to aerobic regulation was analysed in Staphylococcus carnosus. Nitrate induction of a narG-lip reporter gene required presence of NreB/NreC. When nreA was deleted, nitrate was no longer required for maximal induction, suggesting that NreA is a nitrate regulated inhibitor of NreB/NreC. In vitro, NreA and mutant NreA(Y95A) decreased NreB phosphorylation in part or completely, which was due to the inhibition of the autophosphorylating activity rather than an increase of phosphatase activity. Inhibition of phosphorylation was relieved completely when the nitrate-bound NreA was used instead of the nitrate-free form. In the bacterial two-hybrid BACTH system and HPINE interaction assays, NreA interacted with NreB, but not with NreC, and the interaction was diminished by nitrate. In summary, NreA interacts with NreB and controls its phosphorylation level in a nitrate dependent manner. In this way nitrate and NreA modulate the function of the oxygen sensor NreB, resulting in nitrate/oxygen co-sensing by an NreA/NreB sensor unit as part of the NreABC-system.

Developmental stage- and concentration-specific sodium nitroprusside application results in nitrate reductase regulation and the modification of nitrate metabolism in leaves of Medicago truncatula plants

Nitric oxide (NO) is a bioactive molecule involved in numerous biological events that has been reported to display both pro-oxidant and antioxidant properties in plants. Several reports exist which demonstrate the protective action of sodium nitroprusside (SNP), a widely used NO donor, which acts as a signal molecule in plants responsible for the expression regulation of many antioxidant enzymes. This study attempts to provide a novel insight into the effect of application of low (100 μΜ) and high (2.5 mM) concentrations of SNP on the nitrosative status and nitrate metabolism of mature (40 d) and senescing (65 d) Medicago truncatula plants. Higher concentrations of SNP resulted in increased NO content, cellular damage levels and reactive oxygen species (ROS) concentration, further induced in older tissues. Senescing M. truncatula plants demonstrated greater sensitivity to SNP-induced oxidative and nitrosative damage, suggesting a developmental stage-dependent suppression in the plant’s capacity to cope with free oxygen and nitrogen radicals. In addition, measurements of the activity of nitrate reductase (NR), a key enzyme involved in the generation of NO in plants, indicated a differential regulation in a dose and time-dependent manner. Furthermore, expression levels of NO-responsive genes (NR, nitrate/nitrite transporters) involved in nitrogen assimilation and NO production revealed significant induction of NR and nitrate transporter during long-term 2.5 mM SNP application in mature plants and overall gene suppression in senescing plants, supporting the differential nitrosative response of M. truncatula plants treated with different concentrations of SNP.

Effects of salicylic acid on seedling growth and nitrogen use efficiency in cucumber (Cucumis sativus L.)

Salicylic acid (SA) is involved in the regulation of metabolic activity and acts as a defense mechanism in plants under various stress conditions. This study was conducted to determine the effects of SA (10–500 µM) on seedling growth, development, and nitrogen use efficiency (NUE) in cucumber (Cucumis sativus L.) plants with and without nitrogen nutrient. SA increased the contents of chlorophyll, total non-structural carbohydrate, and total nitrogen, as well as nitrate assimilation through the induction of nitrate reductase (EC 1.6.6.1) activity in isolated cucumber cotyledons. Accumulation of SA was twice as high in cotyledons without nitrate supply as in those with nitrate supply. The germination percentage, root length, shoot length, and dry mass of cucumbers increased significantly in response to 50 µM SA. Higher concentrations of SA were, however, found to inhibit these physiological activities. The present results show that optimal physiological concentrations of SA (50 µM) are required to increase NUE in field applications, particularly during germination and seedling growth.

Systemic induction of a Capsicum chinense nitrate reductase by the infection with Phytophthora capsici and defence phytohormones

The mRNA differential display technique was used to identify genes from Habanero pepper (Capsicum chinense Jacq.) seedlings whose expression is modified systemically by infection with the oomycete Phytophthora capsici L. Experiments with different oligonucleotide primer combinations revealed that no single gene was synthesised de novo. Instead, the quantitative accumulation of multiple transcripts was found. From these transcripts, levels of a nitrate reductase (Capsicum chinense nitrate reductase, CcNR), which has a high percentage of identity with other Solanaceae NRs, showed a consistent increase a few hours after inoculation (hai) with P.capsici. Reverse northern blotting revealed the existence of basal levels of CcNR transcripts in different adult tissues; however, systemic levels rose dramatically after spraying seedlings with salicylic acid (SA) and ethephon (ET) but not with methyl jasmonate (MeJa). Both P.capsici and defence phytohormones (DP) also modified NR enzymatic activity (nitrite:NAD(+) oxidoreductase; EC 1.7.1.1) with similar kinetics. Because the application of DP induced and activated the CcNR differentially, it is possible that the activity of CcNR is related to a specific host defence response.

Yield Response of Uniculm Wheat (Triticum aestivum L.) to Early and Late Application of Nitrogen: Flag Leaf Development and Senescence

Nitrogen (N) supply increased flag leaf area by increasing cell number. N promoted initiation and development of floret primordia. N availability upto ear emergence affected fertile floret number more than spikelet number and increased the number of grains. Late application of nitrogen at 5 days after anthesis increased percentage nitrogen in grains, grain weight and grain number due to its effect on ability of floret to set grain. Larger number of florets and grains provided sink not only for carbon but also for nitrogen accumulation with high nitrogen availability. The accumulation of grain protein depended on the accumulation and partitioning of reduced N accumulated during the vegetative stage of growth and the relative contributions of nitrate assimilation and N redistribution during grain development in low as well as high nitrogen supply. Flag leaf of ‘Gigas’ wheat retained the ability to synthesize RuBPCO on induction of nitrate reductase by substrate supply at post anthesis stage. The reductions in proteolytic enzymes were coincident with loss of Chl, total soluble protein and nitrate reductase activity from the flag leaves.

Growth and nitrate assimilation in tomato (Solanum lycopersicon) grown with different nitrogen source and treated with cadmium

In tomato seedlings, the effects of Cd on growth and activity of enzymes involved in nitrate assimilation were studied on different nitrogen sources. After one week on basal growth medium, seedlings were transferred and grown for 14 days on a medium containing 5 mM nitrate, 5 mM ammonium or varying ratios of nitrate to ammonium. In each nitrogen medium, plants were divided into control and Cd treated. In nitrate—rich medium, plant dry weigh was improved while growth was more inhibited by Cd stress than in ammonium—rich medium. Regardless of nitrogen source, Cd treatments decreased nitrate contents in leaves and roots. Nitrate reductase and nitrite reductase activities were decreased by presence of ammonium in culture medium. Cd treatments inhibited NR and NiR activities independently of nitrogen form and plant organ. It could be concluded that tomato plants preferred nitrate as a nitrogen source where as external ammonium was found to inhibit growth. The addition of ammonium in nutrient solution slows down Cd absorption. Endogenous nitrate accumulation may mirror an important role of this metabolite in nitrate and nitrite reductase induction.