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Query: EC:1.7.1.2 (nitrate reductase)
 3,861 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We describe the isolation of salt-sensitive Chlamydomonas reinhardtii mutants by insertional mutagenesis using the nitrate reductase (Nit1) gene. The plasmid pMN24, containing Nit1, was used for transformation of 305CW15 (nit1 cw15 mt+), and transformants were selected for complementation of the nit- phenotype. From 6875 nit+ colonies, four transformants (S4, S18, S46, and S66) were isolated that exhibited both Na+ and Li+ sensitivity (sod-), and another transformant (S33) was selected that exhibited sensitivity to Li+ but not Na+ (lit-) based on relative growth comparisons with the wild-type strain. S33, S46, and S66 were no more growth inhibited by sorbitol than was 305CW15. In comparison, S4 and S18 exhibited substantial growth inhibition in medium supplemented with sorbitol. Genetic analyses indicated that the salt-sensitive mutants were each defective in a single recessive gene. The mutant genes in S4 (sod1), S33 (lit1), and S66 (sod3) are linked to a functional copy of Nit1 and are presumably tagged with a pMN24 insertion.
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PMID:Salt-Sensitive Mutants of Chlamydomonas reinhardtii Isolated after Insertional Tagging. 1222 77

The effects of NaCl on the transport rates of cations, NO3-, and reduced N compounds between roots and shoot and on NO3- assimilation rate were examined on plants of two species differing in their sensitivity to salinity, bean (Phaseolus vulgare L. cv Gabriella) and cotton (Gossypium hirsutum L. cv Akala). Biomass production after 20 d in response to 50 and 100 mM NaCl decreased by 48 and 59% in bean, but only 6 and 14% in cotton. The comparison of the flow patterns obtained for control and NaCl-fed plants showed that salinity induced a general decrease in all the fluxes involved in partitioning of N and the various ions. This decrease was markedly higher in bean than in cotton. Within either species, the different flows (uptake, xylem flux, phloem flux) of a given element were affected by NaCl to the same extent with minor exceptions. No specific effect of salinity on any of the components of N partitioning were discerned. The greater sensitivity of nitrate reductase activity to NaCl in bean leaves compared to cotton leaves seems to be due to a decreased compartmentalization of ions rather than to a difference in salt tolerance of the enzyme itself. Overall, our data show that alteration of mineral nutrition is not solely the reflection of a decreased growth rate, but also is a general process that impairs uptake of all the minerals even at mild NaCl salinity.
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PMID:Effects of NaCl on Flows of N and Mineral Ions and on NO3- Reduction Rate within Whole Plants of Salt-Sensitive Bean and Salt-Tolerant Cotton. 1223 96

The inhibition of seedling growth and nitrate reductase activity in 5 d old Vigna radiata (L.) Wilczek cv. Pusa Baisakhi in the presence of 1.0 mM lead acetate increased drastically, if NaCl (6 and 12 EC) was also present in the nutrient media along with the metal salt. Correspondingly higher endogenous Na+ levels were accumulated in the roots and leaves of seedlings in presence of the two stresses. On the other hand, the levels of endogenous lead get reduced in presence of NaCl in both the roots and leaves. Roots accumulated more Pb2+ and Na+ than the leaves. The two stresses affect more drastically in the additive or even synergistic manner during the early growth phase of the seedlings.
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PMID:Effect of lead on growth and nitrate assimilation of Vigna radiata (L.) Wilczek seedlings in a salt affected environment. 1282 Oct 5

The effect of NO3- uptake on cellular pH was studied in maize roots by an in vivo 31P-NMR technique. In order to separate the effects on cytoplasmic pH due to NO3- uptake from those due to NO3- reduction, tungstate was used to inhibit nitrate reductase (NR). The results confirm that in maize roots tungstate inhibited NR activity. 15N-NMR in vivo experiments demonstrated the cessation of nitrogen flux from nitrate to organic compounds. Tungstate affected neither NO3- uptake nor the levels of the main phosphorylated compounds. Slight changes in cytoplasmic pH were observed during NO3- uptake and reduction (i.e. control). By contrast, in the presence of tungstate, a consistent decrease in cytoplasmic pH occurred. The vacuolar pH did not change in any of the conditions tested. These data show that NO3- uptake is an acidifying process and suggest a possible involvement of NO3- reduction in pH homeostasis. In the presence of NO3-, a transient depolarization of transmembrane electric potential difference (Em) was observed in all the conditions analysed. However, in tungstate-treated roots, a lesser depolarization accompanied by a greater ability to recover Em was found. This was related to a higher activity of the plasma membrane (PM) H+-ATPase. When NO3- was administered as potassium salt, its uptake increased and a greater depolarization of Em took place, whilst the changes in cytoplasmic pH were remarkably reduced, according to the central role played by K+ in the control of plasma membrane activities and cell pH homeostasis. A possible involvement of cytoplasmic pH in the control of PM H+-ATPase expression during nitrate exposure is suggested.
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PMID:Effect of NO3- transport and reduction on intracellular pH: an in vivo NMR study in maize roots. 1531 Aug 18

The Haloferax mediterranei nar operon has been sequenced and its regulation has been characterized at transcriptional level. The nar operon encodes seven open reading frames(ORFs) (ORF1 narB, narC, ORF4, narG, narH, ORF7 and narJ). ORF1, ORF4 and ORF7 are open reading frames with no assigned function, however the rest of them encoded different proteins. narB codes for a 219-amino-acid-residue iron Rieske protein. narC encodes a protein of 486 amino acid residues identified by databases searches as cytochrome-b (narC). The narG gene encodes a protein with 983 amino acid residues and is identified as a respiratory nitrate reductase catalytic subunit (narG). NarH protein has been identified as an electron transfer respiratory nitrate reductase subunit (narH). The last ORF encodes a chaperonin-like protein (narJ) of 242 amino acid residues. The respiratory nitrate reductase was purified 21-fold from H. mediterranei membranes. Based on SDS-PAGE and gel-filtration chromatography under native conditions, the enzyme complex consists of two subunits of 112 and 61 kDa. The optimum temperature for activity was 70 degrees C at 3.4 M NaCl and the stability did not show a direct dependence on salt concentration. Respiratory nitrate reductase showed maximum activity at pH 7.9 and pH 8.2 when assays were carried out at 40 and 60 degrees C, respectively. The absorption spectrum indicated that Nar contains Fe-S clusters. Reverse transcriptase (RT-PCR) shows that regulation of nar genes occurs at transcriptional level induced by oxygen-limiting conditions and the presence of nitrate.
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PMID:Respiratory nitrate reductase from haloarchaeon Haloferax mediterranei: biochemical and genetic analysis. 1534 13

The influence of varying levels of salinity (0, 100, 200 and 400 mM) on the activities of nitrate reductase (NR, E.C. 1.6.6.1), acid phosphatase (ACP, E.C. 3.1.3.2), and alkaline phosphatase (ALP, EC 3.1.3.1 ) as well as on nitrate and phosphate uptake and total nitrogen levels in leaves of a true mangrove Bruguiera parviflora was investigated under hydroponic culture conditions. NR activity increased in 100mM NaCl treated plants, whereas it decreased gradually in 200 and 400 mM treated plants, relative to the controls. Decreased activity of NR by NaCl stress was also accompanied by a decrease in total nitrogen level and nitrate uptake. Decreases in NR activity, nitrate (NO3-), and total nitrogen level due to high salinity may be responsible for a decrease in growth and biomass production in this plant. However, salinity caused an increase in both ACP and ALP activity. Activity staining of ACP by native polyacrylamide gel electrophoresis revealed three isoforms: ACP-1, ACP-2, and ACP-3. We observed a preferential enhancement in the ACP-3 isoform by salinity. In order to understand whether the salinity-induced increase in phosphatase activity was due to inhibition in phosphate uptake, we monitored phosphate (Pi) levels in leaves and noted that phosphate levels decreased significantly under salinity. These results suggest that the induction of acid and ALP under salt stress may be due to a phosphorous deficiency.
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PMID:Effects of NaCI stress on nitrogen and phosphorous metabolism in a true mangrove Bruguiera parviflora grown under hydroponic culture. 1538 3

Loss of the Salmonella MsbB enzyme, which catalyzes the incorporation of myristate destined for lipopolysaccharide in the outer membrane, results in a strong phenotype of sensitivity to salt and chelators such as EGTA and greatly diminished endotoxic activity. MsbB- salmonellae mutate extragenically to EGTA-tolerant derivatives at a frequency of 10(-4) per division. One of these derivatives arose from inactivation of somA, which suppresses sensitivity to salt and EGTA. Here we show that a second mode of MsbB- suppression is a RecA-dependent deletion between two IS200 insertion elements present in Salmonella enterica serovar Typhimurium strain ATCC 14028 but not in two other wild-type strains, LT2 and SL1344, which lack one of the IS200 elements. This deletion occurs spontaneously in wild-type and MsbB- strain 14028 salmonellae and accounts for about one-third of all of the spontaneous suppressors of MsbB- in strain 14028. It spans the region corresponding to 17.7 to 19.9 centisomes, which includes somA, on the sequenced map of Salmonella LT2 (136 ORFs in that strain; ATCC 14028 and other strains showed variability in this region). In addition to conferring EGTA resistance correlated with somA, the deletion confers a MacConkey galactose resistance phenotype on MsbB- Salmonella, indicating that at least one additional gene (distinct from somA) within the deletion is responsible for this phenotype. In the wild type, the deletion mutant grows with normal exponential growth rate in Luria broth but is chlorate resistant and does not grow on citrate agar. The deletion strains have lost hydrogen sulfide production, nitrate reductase activity, and gas production from glucose fermentation.
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PMID:Hot spot for a large deletion in the 18- to 19-centisome region confers a multiple phenotype in Salmonella enterica serovar Typhimurium strain ATCC 14028. 1557 2

The salt-tolerant Rhodotorula glutinis yeast strain grew in medium containing nitrate, 1 mM tungsten, and trace amounts of molybdenum (as impurities from the reagents used). Isolation of electrophoretically homogenous preparation of nitrate reductase from the Rh. glutinis cells grown under these growth conditions is described. The isolated nitrate reductase is a molybdenum-containing homodimer with molecular mass of 130 kD, containing 0.177 mol of Mo per mol of the enzyme. The activity of the enzyme is maximal at pH 7.0 and 35-45 degrees C and is inhibited by low concentrations of azide and cyanide. The enzyme is almost insensitive to 1 mM tungsten.
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PMID:Isolation, purification, and characterization of nitrate reductase from a salt-tolerant Rhodotorula glutinis yeast strain grown in the presence of tungsten. 1609 46

The results showed that when Thellungiella halophila was treated with NaCl, the fresh and dry weight, the water content, the succulency of whole plant and the root/shoot ratio were decreased (Figs. 2-4, 7); the organic matter content in roots was increased and the inorganic matter content in roots was decreased, while those in shoots changed in the opposite direction (Fig. 6); osmotic adjustment ability, the Na+ content, the root activity were increased (Figs. 5, 7, 8); the nitrate reductase activity increased significantly; the O(-)(2*) content decreased at about NaCl 50 mmol/L but increased at about NaCl 100-400 mmol/L (Fig. 10). The micrographs of T. halophila leaf surface by scanning electron microscope (SEM) indicate that there is no salt gland or bladder on the surface of T. halophila (Fig. 1), so it is not a salt-secreting halophyte. The determination of growth parameters, the Na(+) content and Na(+) X-ray (Table 1) microanalysis of T. halophila indicate that T. halophila is not a salt-exclusing halophyte but it probability is a salt-dilution halophyte.
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PMID:[Effects of salt stress on the growth and the nitrate reductase activity in Thellungiella halophila]. 1622 88

Phototrophic growth of the moderate halotolerant Rhodobacter capsulatus strain E1F1 in media containing up to 0.3 M NaCl was dependent on the nitrogen source used. In these media, increased growth rates and growth levels were observed in the presence of reduced nitrogen sources such as ammonium and amino acids. When the medium contained an oxidized nitrogen source (dinitrogen or nitrate), increases in salinity severely inhibited phototrophic growth. However, the addition of glycine betaine promoted halotolerance and allowed the cells to grow in 0.2 M NaCl. Inhibition of diazotrophic growth by salinity was due to a decrease in nitrogenase activity which was no longer synthesized and reversibly inactivated, both effects being alleviated by the addition of glycine betaine. In R. capsulatus E1F1, inhibition of cell growth in nitrate by salt was due to a rapid inhibition of nitrate uptake, which led to a long-term decrease in nitrate reductase activity, probably caused by repression of the enzyme. Addition of glycine betaine immediately restored nitrate uptake, but the recovery of nitrate reductase activity required several hours. Neither ammonium uptake nor ammonium assimilation through the glutamine synthetase-glutamate synthase pathway was affected by NaCl.
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PMID:Halotolerance of the Phototrophic Bacterium Rhodobacter capsulatus E1F1 Is Dependent on the Nitrogen Source. 1653 98


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