Nitrate Utilization Research Articles

FIP1 Plays an Important Role in Nitrate Signaling and Regulates CIPK8 and CIPK23 Expression in Arabidopsis.

Unraveling the molecular mechanisms of nitrate regulation and deciphering the underlying genetic network is vital for elucidating nitrate uptake and utilization in plants. Such knowledge could lead to the improvement of nitrogen-use efficiency in agriculture. Here, we report that the FIP1 gene (factor interacting with poly(A) polymerase 1) plays an important role in nitrate signaling in Arabidopsis thaliana. FIP1 encodes a putative core component of the polyadenylation factor complex. We found that FIP1 interacts with the cleavage and polyadenylation specificity factor 30-L (CPSF30-L), which is also an essential player in nitrate signaling. The induction of nitrate-responsive genes following nitrate treatment was inhibited in the fip1 mutant. The nitrate content was also reduced in fip1 seedlings due to their decreased nitrate uptake activity. Furthermore, the nitrate content was higher in the roots but lower in the roots of fip1, which may result from the downregulation of NRT1.8 and the upregulation of the nitrate assimilation genes. In addition, qPCR analyses revealed that FIP1 negatively regulated the expression of CIPK8 and CIPK23, two protein kinases involved in nitrate signaling. In the fip1 mutant, the increased expression of CIPK23 may affect nitrate uptake, resulting in its lower nitrate content. Genetic and molecular evidence suggests that FIP1 and CPSF30-L function in the same nitrate-signaling pathway, with FIP1 mediating signaling through its interaction with CPSF30-L and its regulation of CIPK8 and CIPK23. Analysis of the 3′-UTR of NRT1.1 showed that the pattern of polyadenylation sites was altered in the fip1 mutant. These findings add a novel component to the nitrate regulation network and enhance our understanding of the underlying mechanisms for nitrate signaling.

Laboratory growth of denitrifying water column microbial consortia from deep-sea shipwrecks in the northern Gulf of Mexico

Background: Shipwrecks serve as a rich source for novel microbial populations that have largely remained undiscovered. Low temperatures, lack of sunlight, and the availability of substrates derived from the shipwreck’s hull and cargo may provide an environment in which microbes can develop unique metabolic adaptations. Methods: To test our hypothesis that shipwrecks could influence the microbial population involved in denitrification when a consortium is grown in the laboratory, we collected samples proximate to two steel shipwrecks in the northern Gulf of Mexico. Then under laboratory conditions, we grew two independent denitrifying microbial consortia. Each consortium was grown by using the BART assay system and analyzed based on growth kinetics, ion chromatography and 16S amplicon sequencing. Results: Both denitrifying consortia were different from each other based on varied growth profiles, rates of nitrate utilization and 16S amplicon sequencing. Conclusions: Our observations conclude that the laboratory grown water column microbial consortia from deep-sea shipwrecks in the Gulf of Mexico are able to undergo aggressive denitrification.

ArgR of Streptomyces coelicolor Is a Pleiotropic Transcriptional Regulator: Effect on the Transcriptome, Antibiotic Production, and Differentiation in Liquid Cultures.

ArgR is a well-characterized transcriptional repressor controlling the expression of arginine and pyrimidine biosynthetic genes in bacteria. In this work, the biological role of Streptomyces coelicolor ArgR was analyzed by comparing the transcriptomes of S. coelicolor ΔargR and its parental strain, S. coelicolor M145, at five different times over a 66-h period. The effect of S. coelicolor ArgR was more widespread than that of the orthologous protein of Escherichia coli, affecting the expression of 1544 genes along the microarray time series. This S. coelicolor regulator repressed the expression of arginine and pyrimidine biosynthetic genes, but it also modulated the expression of genes not previously described to be regulated by ArgR: genes involved in nitrogen metabolism and nitrate utilization; the act, red, and cpk genes for antibiotic production; genes for the synthesis of the osmotic stress protector ectoine; genes related to hydrophobic cover formation and sporulation (chaplins, rodlins, ramR, and whi genes); all the cwg genes encoding proteins for glycan cell wall biosynthesis; and genes involved in gas vesicle formation. Many of these genes contain ARG boxes for ArgR binding. ArgR binding to seven new ARG boxes, located upstream or near the ectA-ectB, afsS, afsR, glnR, and redH genes, was tested by DNA band-shift assays. These data and those of previously assayed fragments permitted the construction of an improved model of the ArgR binding site. Interestingly, the overexpression of sporulation genes observed in the ΔargR mutant in our culture conditions correlated with a sporulation-like process, an uncommon phenotype.

Acute ingestion of beetroot juice increases exhaled nitric oxide in healthy individuals.

Background and objectiveNitric oxide (NO) plays an important role in the airways’ innate immune response, and the fraction of exhaled NO at a flow rate of 50mL per second (FENO50) has been utilized to capture NO. Deficits in NO are linked to loss of bronchoprotective effects in airway challenges and predict symptoms of respiratory infection. While beetroot juice supplements have been proposed to enhance exercise performance by increasing dietary nitrate consumption, few studies have examined the impact of beetroot juice or nitrate supplementation on airway NO in contexts beyond an exercise challenge, which we know influences FENO50.MethodsWe therefore examined the influence of a beetroot juice supplement on FENO50 in healthy males and females (n = 38) during periods of rest and in normoxic conditions. FENO50, heart rate, blood pressure, and state affect were measured at baseline, 45 minutes, and 90 minutes following ingestion of 70ml beetroot juice (6.5 mmol nitrate). Identical procedures were followed with ingestion of 70ml of water on a control day.ResultsAfter beetroot consumption, average values of the natural log of FENO50 (lnFENO50) increased by 21.3% (Cohen’s d = 1.54, p < .001) 45 minutes after consumption and by 20.3% (Cohen’s d = 1.45, p < .001) 90 min after consumption. On the other hand, only very small increases in FENO50 were observed after consumption of the control liquid (less than 1% increase). A small subset (n = 4) of participants completed an extended protocol lasting over 3 hours, where elevated levels of FENO50 persisted. No significant changes in cardiovascular measures were observed with this small single dose of beetroot juice.ConclusionAs NO serves a key role in innate immunity, future research is needed to explore the potential clinical utility of beetroot and dietary nitrate to elevate FENO50 and prevent respiratory infection.

Multiple oxidation state trace elements in suboxic waters off Peru: In situ redox processes and advective/diffusive horizontal transport

Abstract Oxygen minimum zones (OMZs), where the concentrations of oxygen are at analytical detection limits, but sulfate reduction is not occurring, are termed suboxic and typically nitrate serves as the terminal electron acceptor for heterotrophic respiration. Such respiration is highest near the top of OMZs where maxima in nitrite and other redox active species are observed. Within OMZs, the availability of free electrons for reduction reactions (pe) can range from − 2 to 10 and at thermodynamic equilibrium could control the oxidation states of essential trace elements like iron and toxic ones like arsenic. In turn, this oxidation/reduction speciation affects trace element solubility and bioavailability , thus controlling their reactivity and transport. Field observations and mesoscale models demonstrate the increasing areal extent of oxygen minimum zones, but whether this in turn affects trace element cycling and fluxes remains to be verified. From current observations, it is difficult to determine the relative importance of in situ processes and lateral transport from boundaries to trace elements' redox cycling, which are typically highly redox- and biologically-active in these systems. Such processes could lead to the co-occurrence of redox species that are not controlled by a unique pe. In order to evaluate the effects of in situ redox processes versus advective/diffusive transport on trace elements, the cycling of nitrate/nitrite, iodate/iodide, Fe(II), As(III)/As(V), and hydrogen sulfide was examined in the low oxygen waters off the Peru shelf to the open South Pacific Ocean during the US GEOTRACES GP16 transect in 2013. Nitrite, Fe(II), and iodide were observed from the shelf to 95° W, while reduced forms of sulfur and arsenic were absent. Maxima in nitrite, Fe(II) and iodide were coincident, indicating the utilization of Fe(III), iodate and nitrate as terminal electron acceptors was possible. For iron, this finding was unexpected with regards to thermodynamics. However, closer inspection combined with advective/diffusive modeling of water column data and 228Ra-based flux calculations, showed that benthic redox processes, coupled with westward isopycnal transport, influenced the chemical composition and redox speciation within the upper OMZ well offshore. This horizontal transport contributed to the coexistence of Fe(II) with nitrate, indicating that Fe redox chemistry is under kinetic, rather than thermodynamic, control.

Poly-β-hydroxybutyrate (PHB) production by Synechocystis PCC6803 from CO2: Model development

The biosynthesis of poly-β-hydroxybutyrate (PHB) by bioconversion of CO2 is a sustainable alternative to the non-renewable, petroleum-based polymer production. Indeed, the PHB production by conversion of CO2 contributes to the reduction of the greenhouse-gas concentration in the atmosphere. A kinetic dynamic model of PHB production by autotrophic cultures of Synechocystis PCC6803 was proposed and developed by means of the biochemical networks simulator COPASI.Two classes of cells were assumed to be present in the broth: growing cells, PHB producing cells. The model included the two classes of cells and their nitrogen and phosphate internal quota. The dynamics of the cell growth and PHB production were described taking into account: cellular growth rate; lysis rate; nitrate and phosphate utilization rate; PHB production rate. The assessment of the kinetic parameters and of the yields (model calibration) was carried out by the regression of experimental data. Tests were carried out in photobioreactors under dynamic light system (light/dark cycle) using media characterized by initial nitrate concentration ranging between 0 and 1.5g/L. The developed model was validated with respect to independent experimental set.The proposed model successfully reproduced the experimental data (cell concentration, nitrogen and phosphate concentration and PHB content): the square correlation coefficient of the investigated variable concentrations ranged between 0.81 and 0.99. Parameter sensitivity analysis was carried out to assess the role of the selected parameters on cell growth and PHB accumulation. The dynamics of cellular growth were not significantly affected by a ±20% variation of maximum specific growth rate, of velocity of conversion to PHB producing cells, and of maximum uptake rate of nitrate. The PHB accumulation dynamics were particularly sensitive to the variation of the value of the investigated parameters.The proposed model may support the design and the optimization of a PHB production process by means of autotrophic cultures.

Nitrogen Removal From Municipal Wastewater Using a Two‐Sludge Denitrification/Nitrification Batch Reactor: Performance and Mechanisms

Biosorption is a common phenomenon, in that organics in wastewater are trapped by sludge flocs. To achieve high nitrogen removal efficiency from the treatment of low chemical oxygen demand (COD) content wastewater, a two‐sludge denitrification/nitrification batch reactor (TDNBR) is successfully established based on the sorption of organic matter by anoxic activated sludge. During 123 days of operation, the TDNBR achieved high ammonia (&gt;99%) and total nitrogen (TN) (80%) removal efficiency. A higher C/N ratio and exchange frequency have no obvious effect on nitrification but effectively enhanced denitrification. The sorption assays indicated that anoxic sludge could take up a large amount of organic matter (23.79–55.62 mg COD g−1 TSS) but has a low sorption capacity of NH4+‐N. Nitrate utilization rate tests reveal that both the soluble and colloidal organics pre‐sorbed on the sludge can be utilized in denitrification with few organics stored as PHA in bacterial cells. Through cyclic study analysis, it was found that, by interchanging the supernatant in oxic and anoxic tanks alternately, nitrification and denitrification processes are successfully separated and high ammonia and TN removal efficiencies are achieved.

Laboratory growth of denitrifying water column microbial consortia from deep-sea shipwrecks in the northern Gulf of Mexico.

Shipwrecks serve as a rich source for novel microbial populations that have largely remained undiscovered. Low temperatures, lack of sunlight, and the availability of substrates derived from the shipwreck's hull and cargo may provide an environment in which microbes can develop unique metabolic adaptations. Methods: To test our hypothesis that shipwrecks could influence the microbial population involved in denitrification when a consortium is grown in the laboratory, we collected samples proximate to two steel shipwrecks in the northern Gulf of Mexico. Then under laboratory conditions, we grew two independent denitrifying microbial consortia. Each consortium was grown by using the BART assay system and analyzed based on growth kinetics, ion chromatography and 16S amplicon sequencing. Both denitrifying consortia were different from each other based on varied growth profiles, rates of nitrate utilization and 16S amplicon sequencing. Our observations conclude that the laboratory grown water column microbial consortia from deep-sea shipwrecks in the Gulf of Mexico are able to undergo aggressive denitrification.

Nanocarbon coating on the basis of partially reduced graphene oxide

There has been developed an approach to the production of graphene as a result of the thermal reduction of graphene oxide (GO). GO has been synthesized by the use of the modified Hummers method with utilization of sodium nitrate and concentrated sulfuric acid. A paper-like material of 40 – 60 μm in thickness and 1.2 g/cm3 in density was formed on a filter after deposition from dispersion. The material was cut onto samples of about 15×25 mm2 in size which were experienced to the thermal treatment at various temperatures between 100 and 800 °C. This resulted in a set of GO samples reduced to various degrees. The degree of reduction was determined on the basis of measurements of the conductivity. Along with that the evolution of samples density was studied as the annealing temperature was enhanced. The analysis of the X-ray photoelectron spectra of partially reduced GO permitted the determination of the dynamics of changing the chemical composition of the material in the process of the thermal treatment. The analysis of Raman spectra of the GO samples indicates rather high degree of the disordering of the material. A possibility of the usage of the material produced as a nanocarbon coating in experiments on the interaction of high intense liquid flows with a wall surface is discussed.

ADAPTATION OF THE ACTIVATED SLUDGE TO THE DIGESTATE LIQUORS DURING THE NITRIFICATION AND DENITRIFICATION PROCESSES

The activated sludge process of the digestate liquors after chemical separation was conducted using a 10 L lab-scale sequencing batch reactor (SBR) and a 0.50 m3 pilotscale SBR independently (with pH control). Due to the relatively high concentration of free ammonia (FA), clear inhibitory effects of the digestate liquors on the nitrifying bacteria were observed. The adaptation of the activated sludge to the toxicity was evaluated with the trends of ammonia uptake rate (AUR) and nitrate utilization rate (NUR). The lab-scale AUR values decreased from 5.3 to 2.6 g N/(kg VSS·h) over time after the addition of digestate liquors (5–10% of the reactor working volume), indicating an apparent FA inhibition on the nitrification process in the FA concentration range of 0.3–0.5 mg N/L. The pilot-scale AUR values increased from 1.8 to 3.6 g N/ (kg VSS·h) in the first two weeks and then decreased to 2.4 g N/(kg VSS·h), showing a lag of the inhibition on the nitrifying bacteria at the FA concentration ≈ 0.15 mg N/L. The lab-scale NURs increased from 2.6 to 10.4 g N/(kg VSS·h) over time, and the pilot-scale NURs increased from 1.0 to 4.0 g N/(kg VSS·h) in a similar pattern. The clear dependence of both the lab- and pilot-scale NURs on time indicated the adaptation of the heterotrophic biomass to the digestate liquors. Ethanol – used instead of fusel oil – was found to be a more efficient external carbon source for better adaptation of the activated sludge under unfavorable conditions.

NtrC-dependent control of exopolysaccharide synthesis and motility in Burkholderia cenocepacia H111.

Burkholderia cenocepacia is a versatile opportunistic pathogen that survives in a wide variety of environments, which can be limited in nutrients such as nitrogen. We have previously shown that the sigma factor σ54 is involved in the control of nitrogen assimilation and virulence in B. cenocepacia H111. In this work, we investigated the role of the σ54 enhancer binding protein NtrC in response to nitrogen limitation and in the pathogenicity of H111. Of 95 alternative nitrogen sources tested the ntrC showed defects in the utilisation of nitrate, urea, L-citrulline, acetamide, DL-lactamide, allantoin and parabanic acid. RNA-Seq and phenotypic analyses of an ntrC mutant strain showed that NtrC positively regulates two important phenotypic traits: exopolysaccharide (EPS) production and motility. However, the ntrC mutant was not attenuated in C. elegans virulence.

無菌, 非無菌下における二十日大根のNH_4-N, NO_3-Nの利用と体内遊離アミノ酸, アマイド含量

無菌, 非無菌下における二十日大根のNH_4-N, NO_3-Nの利用と体内遊離アミノ酸, アマイド含量

Relationship between δ15N values of bulk sediments and total organic carbon concentration in response to orbital-scale biogenic opal production in the Bering slope area over the last 600 kyrs

Biogenic opal production in the surface water of the Bering Sea shows clear orbital-scale variations such as interglacial highs and glacial lows. In this study, we examined the variation patterns in δ15N of bulk sediment (δ15Nbulk) reflecting nitrate utilization, and total organic carbon (TOC) concentration in terms of organic carbon (OC) production, in response to the orbital-scale biogenic opal production during the last 600 kyrs at Site U1343 (IODP Exp. 323) in the Bering slope area. At Site U1343, δ15Nbulk value and TOC concentration do not show the distinct glacial-interglacial change. Moreover, contrary to the weak correlation between TOC and biogenic opal concentrations, TOC concentration is positively correlated with δ15Nbulk value, indicating the strong control of OC production on nitrate utilization. It should be noted that glacial terminations are characterized by high δ15Nbulk value, high TOC and low biogenic opal concentration whereas the following full interglacials are characterized by low δ15Nbulk value, low TOC and high biogenic opal concentration during the last 600 kyrs. In other words, glacial terminations are characterized by complete nitrate utilization associated with high OC production under low biogenic opal production, whereas the following full interglacial periods are characterized by decreased nitrate utilization associated with low OC production under high biogenic opal production. Such different glacial-interglacial variation patterns of OC production and nitrate utilization in response to biogenic opal production can best be explained by variable Si/C (or Si/N) uptake ratio by diatoms possibly owing to Fe availability. The glacial Fe input increase at Site U1343 is thought to be ascribed to more dust input or large discharge from the glacial Yukon River. These findings imply that Fe transport and contribution may play a significant role in governing the nutrient cycles in the Bering slope region of the Northern Hemisphere.

Nitrification, denitrification, and dephosphatation capability of activated sludge during co-treatment of intermediate-age landfill leachates with municipal wastewater

ABSTRACTThis study focuses on the possible use and efficacy of the co-treatment of landfill leachate (intermediate-age) with municipal wastewater. The nitrification, denitrification, and dephosphatation capability of activated sludge acclimated with a mixture of raw municipal wastewater (RWW) with gradually increasing amounts of raw landfill leachate (RLL) (from 0.5 to 5% v/v) were tested. Biochemical tests were conducted simultaneously in batch reactors (BRs). According to the obtained data, the ammonia utilization rate (AUR) was 3.68 g N/(kg volatile suspended solids (VSS)·h) for RWW, and it increased to 5.78 g N/(kg VSS·h) with the addition of 5% RLL. The nitrate utilization rate under anoxic conditions (NURAX) remained at a comparable level of 1.55–1.98 g N/(kg VSS·h). During the anoxic phase, both nitrate utilization and phosphorus uptake occurred, suggesting that denitrifying phosphorus-accumulating organisms (DPAOs) utilized N–NO3. With the addition of RLL, the rates of anoxic and aerobic phosphate uptake (PURAX and PURAE) and phosphate release rate (PRR) decreased. The PRR was likely negatively influenced by high N–NO3 concentrations but not completely inhibited due to the availability of a biodegradable fraction of chemical oxygen demand (COD). Thus, monitoring the NH4–N load in wastewater treatment plant influent before co-treatment is more informative than that using hydraulic-based criteria.Abbreviations: σ – standard deviation; AUR – ammonia utilization rate; DPAO – denitrifying phosphorus-accumulating organisms; MLVSS – mixed liquor volatile suspended solids content; MSW – municipal solid waste; NURAE – nitrate production rate under aerobic conditions; NURAX – nitrate utilization rate under anoxic conditions; PAO – phosphorus-accumulating organisms; PRR – phosphate release rate under anaerobic conditions; PURAX – phosphate uptake rate under anoxic conditions; PURAE – phosphate uptake rate under aerobic conditions; RLL – raw landfill leachates; RM0.5 – raw mixture of RWW with 0.5% (vol.) of RLL; RM3 – raw mixture of RWW with 3% (vol.) of RLL; RM5 – raw mixture of RWW with 5% (vol.) of RLL; RWW – wastewater; VFA – volatile fatty acids; WWTP – wastewater treatment plant

Enhancement of antibiotic productions by engineered nitrate utilization in actinomycetes.

Nitrate is necessary for primary and secondary metabolism of actinomycetes and stimulates the production of a few antibiotics, such as lincomycin and rifamycin. However, the mechanism of this nitrate-stimulating effect was not fully understood. Two putative ABC-type nitrate transporters were identified in Streptomyces lincolnensis NRRL2936 and verified to be involved in lincomycin biosynthesis. With nitrate supplementation, the transcription of nitrogen assimilation genes, nitrate-specific ABC1 transporter genes, and lincomycin exporter gene lmrA was found to be enhanced and positively regulated by the global regulator GlnR, whose expression was also improved. Moreover, heterologous expression of ABC2 transporter genes in Streptomyces coelicolor M145 resulted in an increased actinorhodin production. Further incorporation of a nitrite-specific transporter gene nirC, as in nirC-ABC2 cassette, led to an even higher actinorhodin production. Similarly, the titers of salinomycin, ansamitocin, lincomycin, and geldanamycin were increased with the integration of this cassette to Streptomyces albus BK3-25, Actinosynnema pretiosum ATCC31280, S. lincolnensis LC-G, and Streptomyces hygroscopicus XM201, respectively. Our work expanded the nitrate-stimulating effect to many antibiotic producers by utilizing the nirC-ABC2 cassette for enhanced nitrate utilization, which could become a general tool for titer increase of antibiotics in actinomycetes.

Millennial-scale changes of surface and bottom water conditions in the northwestern Pacific during the last deglaciation

Changes in water column conditions in the northwestern Pacific during the last 23ka were reconstructed using geochemical and isotope proxies and redox elemental compositions along with published data (alkenone sea surface temperature (SST) and benthic foraminiferal fauna) at core GH02-1030. Surface water primary productivity in terms of biogenic opal and TOC contents, which mainly represented export production of diatom, was closely related to alkenone (spring-summer) SST and the development of spring-summer mixed layer depth. The different variation patterns of nitrate and silicic acid utilization, estimated by bulk δ15N and δ30Sidiatom values, respectively, are most likely due to the water column denitrification influence on bulk δ15N. Dysoxic bottom water conditions occurred during the Bølling-Allerød (BA) and the Pre-Boreal (PB), which was evident by laminated sediments, abundant dysoxic benthic foraminifers, and increased redox elemental compositions. Although surface water productivity increased during the BA and PB, dysoxic bottom water conditions were caused by a combination of enhanced surface water productivity and reduced ventilation of North Pacific Intermediate Water (NPIW) in response to meltwater input from the high latitude areas. Based on records of core GH02-1030 and other cores in the northwestern Pacific, the Okhotsk Sea, and the Bering Sea, which are all proximal to the modern NPIW source region, dissolved oxygen concentrations of bottom water were more depleted during the BA than PB. Such difference was attributed to more sluggish NPIW ventilation due to more meltwater input during the BA than the PB. The opening or closure of the Bering Strait is critical to the direction of meltwater transport to the northwestern Pacific.

Cloning and elucidation of the functional role of apple MdLBD13 in anthocyanin biosynthesis and nitrate assimilation

The plant specific lateral organ boundaries domain (LBD) family of transcription factors (TFs) is involved in many aspects of plant growth and development. In this study, MdLBD13, a nitrate-induced LBD family gene from the apple (Malus × domestica), was isolated and characterized. Overexpression of MdLBD13 repressed anthocyanin biosynthesis by reducing expression of the structural genes associated with the flavonoid pathway. Overexpression ofMdLBD13 also repressed expression of the N–responsive genes that are required for nitrate uptake, transport and assimilation, resulting in reduced nitrate content and nitrate reductase activity in apple calli, as well as in Arabidopsis. Ectopic expression of MdLBD13 also promoted lateral root development in transgenic Arabidopsis. These results demonstrate that MdLBD13acts as a negative regulator in anthocyanin biosynthesis and nitrate utilization.

Nitrate Assimilation in Fusarium fujikuroi Is Controlled by Multiple Levels of Regulation.

Secondary metabolite production of the phytopathogenic ascomycete fungus Fusarium fujikuroi is greatly influenced by the availability of nitrogen. While favored nitrogen sources such as glutamine and ammonium are used preferentially, the uptake and utilization of nitrate is subject to a regulatory mechanism called nitrogen metabolite repression (NMR). In Aspergillus nidulans, the transcriptional control of the nitrate assimilatory system is carried out by the synergistic action of the nitrate-specific transcription factor NirA and the major nitrogen-responsive regulator AreA. In this study, we identified the main components of the nitrate assimilation system in F. fujikuroi and studied the role of each of them regarding the regulation of the remaining components. We analyzed mutants with deletions of the nitrate-specific activator NirA, the nitrate reductase (NR), the nitrite reductase (NiR) and the nitrate transporter NrtA. We show that NirA controls the transcription of the nitrate assimilatory genes NIAD, NIIA, and NRTA in the presence of nitrate, and that the global nitrogen regulator AreA is obligatory for expression of most, but not all NirA target genes (NIAD). By transforming a NirA-GFP fusion construct into the ΔNIAD, ΔNRTA, and ΔAREA mutant backgrounds we revealed that NirA was dispersed in the cytosol when grown in the presence of glutamine, but rapidly sorted to the nucleus when nitrate was added. Interestingly, the rapid and nitrate-induced nuclear translocation of NirA was observed also in the ΔAREA and ΔNRTA mutants, but not in ΔNIAD, suggesting that the fungus is able to directly sense nitrate in an AreA- and NrtA-independent, but NR-dependent manner.

HYDROLYSIS OF PRODUCTS OF UREA DENITRATION OF NITRIC ACID RAFFINATE OF EXTRACTION URANIUM REFINING

The paper deals with the nitric acid extraction of uranium that has one significant drawback – the lack of technology for the utilization of nitric acid from raffinate. In the overwhelming majority of the cases, spent nitric acid is neutralized by an alkaline agent, and the nitrates of alkaline or alkaline-earth metals are discharged to the tailings pond. One of the possible ways of handling raffinates is their carbamide denitration which minimizes the unproductive loss of nitric acid. However, until now, the issue of effective treatment of urea nitrate precipitate (CO(NH2)2·HNO3) has not been solved. The paper contains the results of research aimed at solving the problem of utilization of urea nitrate. Urea nitrate can be processed into ammonium nitrate through acid hydrolysis of urea. The subject of the study is a solution containing 200 g/l urea nitrate and a solution containing 200 g/l urea nitrate and 100 g/l nitric acid. The process is studied in the temperature range 70–150 ºC at atmospheric and elevated pressures using an autoclave. IR spectra of the urea nitrate hydrolysis products confirmed the completeness of the process for 4 hours at 150 ºC and a pressure of 30 atm. The introduction of an equimolar amount of nitric acid into the reaction volume increases the rate of hydrolysis of urea nitrate by a factor of 2 and does not impede the completeness of its transformation into ammonium nitrate. The activation energy of the urea nitrate hydrolysis is 28,9 kJ/mol that is comparable to the activation energy of urea enzymatic (urease) hydrolysis. Urea nitrate hydrolyzate (ammonium nitrate) can be used as a desorption solution in the process of uranium sorption refining of the technology of underground uranium leaching.

CO2-induced alterations in plant nitrate utilization and root exudation stimulate N2O emissions

Atmospheric carbon dioxide enrichment (eCO2) often increases soil nitrous oxide (N2O) emissions, which has been largely attributed to increased denitrification induced by CO2-enhancement of soil labile C and moisture. However, the origin of the N remains unexplained. Emerging evidence suggests that eCO2 alters plant N preference in favor of ammonium (NH4+-N) over nitrate (NO3−-N). Yet, whether and how this attributes to the enhancement of N2O emissions has not been investigated. We conducted a microcosm experiment with wheat (Triticum aestivum L.) and tall fescue (Schedonorus arundinaceus (Schreb.) Dumort.) to examine the effects of eCO2 on soil N2O emissions in the presence of two N forms (NH4+-N or NO3−-N). Results obtained showed that N forms dominated eCO2 effects on plant and microbial N utilization, and thus soil N2O emissions. Elevated CO2 significantly increased the rate and the sum of N2O emissions by three to four folds when NO3−-N, but not NH4+-N, was supplied under both wheat and tall fescue. While enhanced N2O emission was more related to the reduced plant NO3−-N uptake under wheat, it concurred with increased labile C under tall fescue. In the presence of NO3−-N, significantly lower shoot biomass N and 15N, but higher plant biomass C:N ratio, microbial biomass C and N, and/or soil extractable C indicated that eCO2 constrained plant NO3−-N utilization and likely stimulated root exudation. We propose a new conceptual model in which eCO2-inhibition of plant NO3−-N uptake and/or CO2-enhancement of soil labile C enhances the N and/or C availability for denitrifiers and increases the intensity and/or the duration of N2O emissions. Together, these findings indicate that CO2-enhancement of soil N and labile C favors denitrification, suggesting that management of N fertilizers in intensive systems will likely become more challenging under future CO2 scenarios.