Nitrate Transport Research Articles

Root system size and root hair length are key phenes for nitrate acquisition and biomass production across natural variation in Arabidopsis.

The role of root phenes in nitrogen (N) acquisition and biomass production was evaluated in 10 contrasting natural accessions of Arabidopsis thaliana L. Seedlings were grown on vertical agar plates with two different nitrate supplies. The low N treatment increased the root to shoot biomass ratio and promoted the proliferation of lateral roots and root hairs. The cost of a larger root system did not impact shoot biomass. Greater biomass production could be achieved through increased root length or through specific root hair characteristics. A greater number of root hairs may provide a low-resistance pathway under elevated N conditions, while root hair length may enhance root zone exploration under low N conditions. The variability of N uptake and the expression levels of genes encoding nitrate transporters were measured. A positive correlation was found between root system size and high-affinity nitrate uptake, emphasizing the benefits of an exploratory root organ in N acquisition. The expression levels of NRT1.2/NPF4.6, NRT2.2, and NRT1.5/NPF7.3 negatively correlated with some root morphological traits. Such basic knowledge in Arabidopsis demonstrates the importance of root phenes to improve N acquisition and paves the way to design eudicot ideotypes.

The Nitrate Transporter MtNPF6.8 Is a Master Sensor of Nitrate Signal in the Primary Root Tip of Medicago truncatula.

Nitrate is not only an essential nutrient for plants, but also a signal involved in plant development. We have previously shown in the model legume Medicago truncatula, that the nitrate signal, which restricts primary root growth, is mediated by MtNPF6.8, a nitrate transporter. Nitrate signal also induces changes in reactive oxygen species accumulation in the root tip due to changes in cell wall peroxidase (PODs) activity. Thus, it was interesting to determine the importance of the role of MtNPF6.8 in the regulation of the root growth by nitrate and identify the POD isoforms responsible for the changes in POD activity. For this purpose, we compared in M. truncatula a npf6.8 mutant and nitrate insensitive line deficient in MtNPF6.8 and the corresponding wild and sensitive genotype for their transcriptomic and proteomic responses to nitrate. Interestingly, only 13 transcripts and no protein were differently accumulated in the primary root tip of the npf6.8-3 mutant line in response to nitrate. The sensitivity of the primary root tip to nitrate appeared therefore to be strongly linked to the integrity of MtNPF6.8 which acts as a master mediator of the nitrate signal involved in the control of the root system architecture. In parallel, 7,259 and 493 genes responded, respectively, at the level of transcripts or proteins in the wild type, 196 genes being identified by both their transcript and protein. By focusing on these 196 genes, a concordance of expression was observed for most of them with 143 genes being up-regulated and 51 being down-regulated at the two gene expression levels. Their ontology analysis uncovered a high enrichment in POD genes, allowing the identification of POD candidates involved in the changes in POD activity previously observed in response to nitrate.

Impact of microplastics from polyethylene and biodegradable mulch films on rice (Oryza sativa L.)

Extensive use of plastic film mulch has resulted in accumulation of a large amount of residual plastic, which will eventually fragment into microplastics in agricultural soils. However, it is unclear how microplastics from plastic mulch film affect crops. To address this issue, rice plants exposed to microplastics derived from poly(butyleneadipate-co-terephthalate) (PBAT)-based biodegradable mulch film (BM) and polyethylene (PE) mulch film (PM) were investigated for plant growth, physio-biochemical processes, and gene expressions. Both types of microplastics significantly reduced the height and dry weight of rice plant. Oxidative stress was induced by microplastics in rice shoot and root, with levels of ROS relatively higher under treatment PM than that under treatment BM. Transcriptomic data showed that more genes were down-regulated by treatment PM than that by treatment BM. Genes encoding ammonium and nitrate transporters were down-regulated by both types of microplastics in rice roots at vegetative stage, whereas up-regulated at reproductive stage, as compared to their respective treatment with no microplastics (CK). Similar results regarding phenylpropanoid biosynthesis pathway and lignin content were also observed in rice roots. Net photosynthetic rate and SPAD value were significantly inhibited by treatments BM and PM in rice shoot, and the expression of genes involved in light reaction was reduced at vegetative stage, whereas there were no differences of them at reproductive stage, as compared to their respective treatment CK. Our study suggests that microplastics from BM and PM both affect the growth of rice plants via nitrogen metabolism and photosynthesis. The negative effects imposed by both types of microplastics on rice plant can be mitigated with the growth of plants, and the negative effects of microplastics from PE mulch film on rice plant are relatively stronger than that from the PBAT-based biodegradable film.

Investigation of the direct effect of a precision Ascophyllum nodosum biostimulant on nitrogen use efficiency in wheat seedlings

Reduction in the greenhouse gas (GHG) emissions and nitrogen (N) pollution of ground water by improving nitrogen use efficiency (NUE) in crops has become an intensively investigated research topic in pursuit of a more sustainable future. Although, distinct solutions have been proposed there are only a few reports documenting the detailed interplay between observed plant growth dynamics and changes in plant N related transcriptional and biochemical changes. It was previously demonstrated that the application of a formulated biostimulant (PSI-362) derived from Ascophyllum nodosum (ANE) improves N uptake in Arabidopsis thaliana and in barley. In this study, the effect of PSI-362 on the growth dynamics of wheat seedlings was evaluated at different biostimulant and N supplementation rates. Wheat grown on N deficient MS medium was also analysed from the first hour of the treatment until the depletion of the nutrients in the medium 9 days later. During this time the biomass increase measured for PSI-362 treated plants versus untreated controls was associated with increased nitrate uptake, with surplus N assimilated by the biomass in the form of glutamate, glutamine, free amino acids, soluble proteins, and chlorophyll. Phenotypical and biochemical analysis were supported by evaluation of differential expression of genetic markers involved in nitrate perception and transport (TaNRT1.1/NPF6.3), nitrate and nitrite reduction (TaNR1 and TaNiR1) and assimilation (TaGDH2, TaGoGAT, TaGS1). Finally, a comparative analysis of the precision biostimulant PSI-362 and two generic ANEs demonstrated that the NUE effect greatly differs depending on the ANE formulation used.

Deep soil nitrogen storage slows nitrate leaching through the vadose zone

Nitrogen (N) fertilizer applications are important for agricultural yield, yet not all the applied N is taken up by crops, leading to surplus N storage in soil or leaching to groundwater and surface water. Leaching loss of fertilizer N represents a cost for farmers and has consequences for human health and the environment, especially in the southern Willamette Valley, Oregon, USA, where groundwater nitrate contamination is prevalent. While improved nutrient management and conservation practices have been implemented to minimize leaching, nitrate levels in groundwater continue to increase in many long-term monitoring wells. To elucidate controls on leaching rates and N dynamics in agricultural soils across soil depths, and in response to seasonal and annual variation in management (e.g., fertilizer input amount and summer irrigation), we intensively monitored the transport of water and nitrate every two weeks for four years through the vadose zone at three depths (0.8, 1.5, and 3.0 m) in a sweet corn (maize) field. Though nitrate leaching was highly variable among lysimeters at the same depth and across years, a strong pattern emerged: annual nitrate leaching significantly decreased with depth across the study, averaging ~104 kg N ha-1 yr-1 near the surface (0.8 m) versus ~56 kg N ha-1 yr-1 in the deep soil (3.0 m), a 54% reduction in leaching between the soil layers. Even though crops were irrigated in summer, most leaching (~72% below 3.0 m) occurred during the wet fall and winter. Based on steady state assumptions, a net equivalent of ~29% of surface N inputs leached below 3.0 m into the deeper soil and groundwater, while ~44% was removed in crop harvest, indicating considerable N retention in the soil (~27% of inputs or approximately 58 kg N ha-1 yr-1). The accumulation and long-term dynamics of deep soil N is a legacy of agricultural management that should be further studied to better manage and reduce nitrate loss to groundwater.

A New Species in Pseudophialophora From Wild Rice and Beneficial Potential.

Wild rice (Oryza granulata) is a natural resource pool containing abundant unknown endophytic fungi species. There are few reports on the endophytic fungi in wild rice. Here, one isolate recovered from wild rice roots was identified as a new species Pseudophialophora oryzae sp. nov based on the molecular phylogeny and morphological characteristics. Fluorescent protein-expressing P. oryzae was used to monitor the fungal colonization pattern. Hyphae invaded the epidermis to the inner cortex but not into the root stele. The inoculation of P. oryzae promoted the rice growth, with the growth parameters of chlorophyll content, shoot height, root length, fresh shoot weight, fresh root weight and dry weight increasing by 24.10, 35.32, 19.35, 90.00, 33.3, and 79.17%, respectively. P. oryzae induced up-regulation of nitrate transporter OsPTR9 and potassium transporter OsHAK16 by 7.28 ± 0.84 and 2.57 ± 0.80 folds, promoting nitrogen and potassium elements absorption. In addition, P. oryzae also conferred a systemic resistance against rice blast, showing a 72.65 and 75.63% control rate in sterile plates and potting conditions. This systemic resistance was mediated by the strongly up-regulated expression of resistance-related genes NAC, OsSAUR2, OsWRKY71, EL5, and PR1α. Since P. oryzae can promote rice growth, biomass and induce systemic disease resistance, it can be further developed as a new biogenic agent for agricultural production, providing a new approach for biocontrol of rice blast.

Global gene expression profiling under nitrogen stress identifies key genes involved in nitrogen stress adaptation in maize (Zea mays L.)

Maize is a heavy consumer of fertilizer nitrogen (N) which not only results in the high cost of cultivation but may also lead to environmental pollution. Therefore, there is a need to develop N-use efficient genotypes, a prerequisite for which is a greater understanding of N-deficiency stress adaptation. In this study, comparative transcriptome analysis was performed using leaf and root tissues from contrasting inbred lines, viz., DMI 56 (tolerant to N stress) and DMI 81 (susceptible to N stress) to delineate the differentially expressed genes (DEGs) under low-N stress. The contrasting lines were grown hydroponically in modified Hoagland solution having either sufficient- or deficient-N, followed by high-throughput RNA-sequencing. A total of 8 sequencing libraries were prepared and 88–97% of the sequenced raw reads were mapped to the reference B73 maize genome. Genes with a p value ≤ 0.05 and fold change of ≥ 2.0 or ≤ − 2 were considered as DEGs in various combinations performed between susceptible and tolerant genotypes. DEGs were further classified into different functional categories and pathways according to their putative functions. Gene Ontology based annotation of these DEGs identified three different functional categories: biological processes, molecular function, and cellular component. The KEGG and Mapman based analysis revealed that most of the DEGs fall into various metabolic pathways, biosynthesis of secondary metabolites, signal transduction, amino acid metabolism, N-assimilation and metabolism, and starch metabolism. Some of the key genes involved in N uptake (high-affinity nitrate transporter 2.2 and 2.5), N assimilation and metabolism (glutamine synthetase, asparagine synthetase), redox homeostasis (SOD, POX), and transcription factors (MYB36, AP2-EREBP) were found to be highly expressed in the tolerant genotype compared to susceptible one. The candidate genes identified in the present study might be playing a pivotal role in low-N stress adaptation in maize and hence could be useful in augmenting further research on N metabolism and development of N-deficiency tolerant maize cultivars.

Nitrate-dependent regulation of miR444-OsMADS27 signalling cascade controls root development in rice.

Nitrate is an important nutrient and a key signalling molecule for plant development. A number of transcription factors involved in the response to nitrate and their regulatory mechanisms have been identified. However, little is known about the transcription factors involved in nitrate sensing and their regulatory mechanisms among crop plants. In this study, we identified functions of a nitrate-responsive miR444:MADS-box transcription factor OsMADS27 module and its downstream targets mediating rice root growth and stress responses. Transgenic rice plants expressing miR444 target mimic improved rice root growth. Although miR444 has the potential to target multiple genes, we identified OsMADS27 as the major miR444 target that regulates the expression of nitrate transporters, as well as several key genes including expansins, and those associated with auxin signalling, to promote root growth. In agreement with this, overexpression of miRNA-resistant OsMADS27 improved root development and tolerance to abiotic stresses, while its silencing suppressed root growth. OsMADS27 mediated robust stress tolerance in plants through its ability to bind to the promoters of specific stress regulators, as observed in ChIP-seq analysis. Our results provide evidence of a nitrate-dependent miR444-OsMADS27 signalling cascade involved in the regulation of rice root growth, as well as its surprising role in stress responses.

Using multiple isotopes to identify sources and transport of nitrate in urban residential stormwater runoff.

Increased nitrogen (N) from urban stormwater runoff aggravates the deterioration of aquatic ecosystems as urbanisation develops. The sources and transport of nitrate (NO3-) in urban stormwater runoff were investigated by analysing different forms of N, water isotopes (δD-H2O and δ18O-H2O), and NO3- isotopes (δ15N-NO3- and δ18O-NO3-) in urban stormwater runoff in a residential area in Hangzhou, China. The results showed that the concentrations of total N and nitrate N in road runoff were higher than those in roof runoff. Moreover, high concentrations of dissolved organic N and particulate N led to high total nitrogen (TN) concentrations in road runoff (mean: 3.76mg/L). The high δ18O-NO3- values (mean: + 60 ± 13.1‰) indicated that atmospheric deposition was the predominant NO3- source in roof runoff, as confirmed by the Bayesian isotope mixing model (SIAR model), contributing 84-98% to NO3-. Atmospheric deposition (34-92%) and chemical fertilisers (6.2-54%) were the main NO3- sources for the road runoff. The proportional contributions from soil and organic N were small in the road runoff and roof runoff. For the initial period, the NO3- contributions from atmospheric deposition and chemical fertilisers were higher and lower, respectively, than those in the middle and late periods in road runoff during storm events 3 and 4, while an opposite trend of road runoff in storm event 7 highlighted the influence of short antecedent dry weather period. Reducing impervious areas and more effective management of fertiliser application in urban green land areas were essential to minimize the presence of N in urban aquatic ecosystems.

Integrative Transcriptome and Proteome Analysis Reveals the Absorption and Metabolism of Selenium in Tea Plants [Camellia sinensis (L.) O. Kuntze

Certain tea plants (Camellia sinensis) have the ability to accumulate selenium. In plants, the predominant forms of bioavailable Se are selenite (SeO32–) and selenate (SeO42–). We applied transcriptomics and proteomics to hydroponically grown plants treated with selenite or selenate for 48 h in the attempt to elucidate the selenium absorption and assimilation mechanisms in tea. A total of 1,844 differentially expressed genes (DEGs) and 691 differentially expressed proteins (DEPs) were obtained by comparing the Na2SeO3 and Na2SeO4 treatments against the control. A GO analysis showed that the genes related to amino acid and protein metabolism and redox reaction were strongly upregulated in the plants under the Na2SeO3 treatment. A KEGG pathway analysis revealed that numerous genes involved in amino acid and glutathione metabolism were upregulated, genes and proteins associated with glutathione metabolism and ubiquinone and terpenoid-quinone biosynthesis were highly expressed. Genes participating in DNA and RNA metabolism were identified and proteins related to glutathione metabolism were detected in tea plants supplemented with Na2SeO4. ABC, nitrate and sugar transporter genes were differentially expressed in response to selenite and selenate. Phosphate transporter (PHT3;1a, PHT1;3b, and PHT1;8) and aquaporin (NIP2;1) genes were upregulated in the presence of selenite. Sulfate transporter (SULTR1;1 and SULTR2;1) expression increased in response to selenate exposure. The results of the present study have clarified Se absorption and metabolism in tea plants, and play an important theoretical reference significance for the breeding and cultivation of selenium-enriched tea varieties.

Transcriptional regulation of metal metabolism- and nutrient absorption-related genes in Eucalyptus grandis by arbuscular mycorrhizal fungi at different zinc concentrations

BackgroundEucalyptus spp. are candidates for phytoremediation in heavy metal (HM)-polluted soils as they can adapt to harsh environments, grow rapidly, and have good economic value. Arbuscular mycorrhizal fungi (AMF) are the most widely distributed plant symbiotic fungi in nature, and they play an important role in promoting the phytoremediation of HM-polluted soils. However, few studies have evaluated the HM detoxification mechanism of E. spp. in symbiosis with AMF, and thus, the molecular mechanism remains unclear.ResultsThe gene transcription and metabolic pathways of E. grandis were studied with and without inoculation with AMF and at different zinc (Zn) concentrations. Here, we focused on the transcript level of six HM-related gene families (ZNT, COPT/Ctr, YSL, ZIFL and CE). Under high-Zn conditions, thirteen genes (ZNT:2, COPT/Ctr:5, YSL:3, ZIFL:1, CE:2) were upregulated, whereas ten genes (ZNT:3, COPT/Ctr:2, YSL:3, ZIFL:1, CE:1) were downregulated. With AMF symbiosis under high-Zn conditions, ten genes (ZNT:4, COPT/Ctr:2, YSL:3, CE:1) were upregulated, whereas nineteen genes (ZNT:9, COPT/Ctr:2, YSL:3, ZIFL:4, CE:1) were downregulated. Under high-Zn conditions, genes of three potassium-related transporters, six phosphate transporters (PHTs), and two nitrate transporters (NRTs) were upregulated, whereas genes of four potassium-related transporters,four PHTs, and four nitrogen-related transporters were downregulated. With AMF symbiosis under high-Zn conditions, genes of two potassium-related transporters, six ammonium transporters (AMTs) and five PHTs were upregulated, whereas genes of six potassium-related transporters, two AMTs and five PHTs were downregulated.ConclusionsOur results indicates that AMF increases the resistance of E. grandis to high-Zn stress by improving nutrients uptake and regulating Zn uptake at the gene transcription level. Meanwhile, our findings provide a genome-level resource for the functional assignments of key genes regulated by Zn treatment and AM symbiosis in six HM-associated gene families and macromineral nutrient-related gene families of E. grandis. This may contribute to the elucidation of the molecular mechanisms of the response to Zn stress in E. grandis with AM symbiosis at the aspect of the interaction between HM tolerance and nutrient acquisition.

Nitrogen Absorption Pattern Detection and Expression Analysis of Nitrate Transporters in Flowering Chinese Cabbage

Nitrate transporters (NRTs) play an important role in nitrate absorption and internal distribution in plant roots and other parts. Experiments were carried out to explore the sequences and expression characteristics of NRT genes, and their correlation with the N uptake in flowering Chinese cabbage. We have isolated three important BcNRTs (BcNRT1.1, BcNRT1.2, and BcNRT2.1) from flowering Chinese cabbage. Spatio-temporal expression analysis found that BcNRT1.1 and BcNRT2.1 were mainly expressed in roots, while BcNRT1.2 was more expressed in roots than in leaves during vegetative growth and was mainly expressed in leaves during reproductive growth. The NO3− uptake rate of the entire growth period was significantly correlated with BcNRT1.1 and BcNRT1.2 expression in roots. In addition, the total N content was increased with the increase in NO3− concentration in flowering Chinese cabbage. The NH4+ uptake was slightly induced by NH4+, but the total N content had no significant difference under the NH4+ concentration of 1–8 mmol/L. We also found that lower concentrations of NH4+ promoted the expression of BcNRT1.1 and BcNRT1.2 while inhibiting the expression of BcNRT2.1 in the roots of flowering Chinese cabbage. The amount of total N uptake in the treatment with 25/75 of NH4+/NO3− was significantly higher than that of the other two treatments (0/100 and 50/50). In the mixture of NH4+ and NO3−, total N uptake was significantly correlated with the BcNRT1.2 expression. We concluded that mixed nutrition with an NH4+/NO3− of 25/75 could significantly increase total nitrogen uptake in flowering Chinese cabbage, in which two members of the NRT1 subfamily (BcNRT1.1 and BcNRT1.2) might play a major regulatory role in it. This study is a beneficial attempt to dig deeper into the NRT genes resources and lays the foundation for the ultimate use of genetic improvement methods to increase the NUE with less nitrogen fertilizer in flowering Chinese cabbage.

Physiological and transcriptome analysis reveals the differences in nitrate content between lamina and midrib of flue-cured tobacco

Nitrate is an important precursor of tobacco-specific nitrosamines (TSNAs) and a remarkable difference in nitrate accumulation between lamina and midrib of flue-cured tobacco has long been observed. However, the physiological and molecular mechanisms underpinning this difference remain poorly understood. In this study, physiological and genetic factors impacting nitrate accumulation were identified in pot experiments using flue-cured tobacco K326 with contrasting nitrate content between lamina and midrib. The results showed that three times higher NO3-N content was observed in midrib than that in the lamina, along with lower pigment, NH4-N content, nitrate reductase activity (NRA), sucrose synthetase activity (SSA), and glutamine synthetase activity (GSA) in midrib. Transcriptome analysis revealed that expression of genes involved in porphyrin and chlorophyll metabolism, carotenoid biosynthesis, photosynthesis-antenna proteins, photosynthesis, carbon fixation in photosynthetic organisms, starch and sucrose metabolism, nitrogen metabolism, and biosynthesis of amino acids were significantly lower in midrib than in lamina. qRT-PCR results showed that the expression level of nitrate transporter genes LOC107782967, LOC107806749, LOC107775674, LOC107829632, LOC107799198, LOC107768465 decreased by 2.74, 1.81, 49.5, 3.5, 2.64 and 2.96-folds while LOC107789301 increased by 8.23-folds in midrib but not in lamina. Reduced chlorophyll content might result in low carbohydrate formation which is the source of energy and carbon skeleton supply, then the low capacity of nitrogen reduction, assimilation and transportation, and the poor ability of nitrate reallocation but the high capacity of accumulation might lead to nitrate accumulation in midrib. The results laid the foundation for reducing nitrate content and TSNA formation in tobacco midribs and their products.

Isolation and Characterization of Bacillus Consortia for Plant Growth Promotion in Rice (Oryza sativa L.)

In this study, soil samples from the rhizosphere of various uncultivated weeds were collected from fifteen different locations of Gujarat. Heat treatment was given at 65°C for 20 minutes prior to initial screening for spore-forming Bacillus spp. Among them, 20 nitrogen-fixing (NFB), 27 phosphate solubilising (PSB) and 15 potassium mobilizing (KMB) isolates were screened primarily. After molecular identification only Bacillus isolates were further selected and characterized. Three superior Bacillus isolates were selected from each category by secondary screening. All isolates belonging to different category were compatible in nature and showed significant ammonia production, ARA, phosphate solubilisation, potassium mobilization, siderophore production, IAA production and organic acid producers. The relative expression analysis of three genes NRT2.1 (Nitrate transporter), PT6 (Phosphorus transporter), and AKT1 (Potassium transporter) at transcriptional level were performed in the juvenile root tissues of Rice using qRT-PCR technique at 15th, 30th and 45th days after transplanting with Actin as a internal control. Rice transplants were inoculated with different NPK fixing Bacillus consortia, which significantly improved growth parameters as compared to control in field study. Various growth and yield attributing characters of rice viz., plant height (at 60 DAT, 90 DAT, and at harvesting), number of productive tillers, days to 50 % flowering, 100 seed weight, grain yield, straw yield, as well as dry biomass per hectare were significantly influenced by different treatments. An application of 100 % RDF with NPK consortia (T7) treatment of rice crop resulted in significantly higher plant height (49.25 cm at 60 DAT, 107.00 cm at 90 DAT, 129.50 cm at harvesting), days to 50% flowering (75.63), 100 seed weight (3.53 g), number of productive tillers per plant (12.13), grain yield (6889.00 kg ha-1), straw yield (8754.25 kg ha-1) and dry biomass (7889.25 kg ha-1) followed by the treatments T4 (T2 + N consortia), T5 (T2 + P consortia), T6 (T2 + K consortia), and T2 (RDF (NP) 100%). Nutrient content in grain and straw was differing significantly due to different treatments. However, significantly higher N, P, K, content in straw (1.28 %, 0.15 %, and 1.52 %) and grain (2.06 %, 0.23 %, and 0.25 %) respectively, were recorded under the treatment T7. The spore forming Bacillus consortia was able to survive at a wide range of temperature and pH fluctuations and found to be effective as N-fixers, P-solubilizers, K-mobilizers, siderophore producers, IAA producers with having antagonistic activity against rice pathogen Magnaportha oryzae.

Evaluating nitrate transport and accumulation in the deep vadose zone of the intensive agricultural region, North China Plain

Evaluation of the nitrate transport process in the deep vadose zone (DVZ) is important for groundwater quality management, especially in intensive agricultural regions, such as the North China Plain (NCP). The NCP produces ~20% of the total food grain in China, owing to timely groundwater irrigation and excessive chemical N fertilizer applications, and faces severe groundwater environmental degradation. This study evaluated the potential impacts of intensive agriculture on groundwater quality by investigating nitrate accumulation and transport in the DVZ of wheat-maize double-cropping field based on sediment sampling (maximum depth of 45.2 m) over three sub-regions of the NCP. The results showed that legacy nitrate‑nitrogen (NO3−-N) accumulated in the DVZ ranged from 118.5 to 6302.8 kg N ha−1 across the NCP; it increased with depth at an average rate of ~157 kg ha−1 m−1. Nitrate transport and accumulation in the DVZ were spatially varied and mainly controlled by the DVZ sediment textures, in addition to water and nitrogen inputs from the ground surface. Coarse sediments retained lower soil water content, resulting in less nitrogen storage; however, they provided greater nitrate transport velocity. Higher transport velocities observed in the alluvial-proluvial fan allowed chemical N fertilizer to reach the water table. However, in other regions, nitrate transport velocities were lower than the water table decline rates, implying that groundwater quality may not have been impaired by chemical N fertilizer. Furthermore, a reductive environment was identified in some areas with fine sediments, indicating a favorable environment for denitrification in the DVZ. The findings of the current study could provide an important foundation for groundwater quality management in agricultural areas, such as the NCP and similar regions.

Nutrient streams in the North Pacific

The nutrient streams in the North Pacific were examined by analyzing the results derived from the inverse calculation of the World Ocean Circulation Experiment (WOCE)-revisit cruise data. The Kuroshio, Alaskan Stream, and Oyashio drive the most intensive nutrient transport within the basin. The upper portion of the nutrient stream (lighter than 26.2 kg/m3) contains mode waters and recirculates within the subtropical gyre, while the intermediate portion (26.2–27.15 kg/m3) relates to North Pacific Intermediate Water (NPIW) and contributes most to nutrient transport in the nutrient stream. The nutrient concentration in the nutrient stream increases in the subtropical gyre along the Kuroshio and Kuroshio Extensions but decreases in the subarctic gyre from the Alaskan Stream to the Oyashio region. This increase or decrease in nutrient concentration from the upstream region to the downstream region mainly results from lateral water exchange, although it also partly depends on biogeochemical processes and vertical diffusion. Specifically, the lateral exchange of the nutrient stream in the subarctic gyre with water from lower latitudes decreases the nitrate concentration of the Alaskan Stream and that with water from the Okhotsk Sea decreases the nitrate concentration of the Oyashio. For vertical nitrate transport, our estimation shows that the ventilation processes of subduction and induction are comparable to the sum of vertical advection and diffusion. This implies that nutrient transport related to the ventilation of the North Pacific is in a dynamic balance on an annual cycle and should be quantified in future work.

Influence of ethanol and nitrate on ibuprofen removal in batch reactors under denitrifying conditions

The optimized removal of the anti-inflammatory drug ibuprofen inoculated with activated sludge biomass in batch reactors was evaluated, using the variables ibuprofen (60 – 80 µg L-1), ethanol (130 – 230 mg EtOH L-1) and nitrate (130 – 230 mg NO3- L-1) by rotational central compound design. A higher removal efficiency of ibuprofen (97.5 ± 3.1%) was observed with 95.9 mg NO3- L-1, 109.9 µg IBU L-1 and 110.6 mg EtOH L-1. Under these conditions, complete removal of NO3- (1.1 h-1 speed), 20 ± 2.7% of organic matter removal (366.5 ± 3.5 mg COD L-1)> 90% ethanol removal were obtained. Based on the mass sequencing of the 16 S rRNA gene via the Illumina platform, bacteria related to the degradation of recalcitrant compounds were identified, such as Pseudomonas (1.5%), Zoogloea (1.73%) and Rhodanobacter (1.63%). The enzymes involved in denitrification were also inferred, such as nitrate transporters and several reductases involved in the conversion of intermediates. Although ibuprofen removal occurred in the absence of ethanol (16.5%) and nitrate (15.2%), the optimal removal range (97.5%) as well as the selection of bacteria possibly involved in its degradation pathways was obtained with 110.6 mg EtOH L-1 and 95.9 mg NO3- L-1, achieving removal approximately 5.9 times higher when compared to the assays without ethanol and nitrate.

Nitrate transport via NRT2.1 mediates NIN-LIKE PROTEIN-dependent suppression of root nodulation in Lotus japonicus.

Legumes have adaptive mechanisms that regulate nodulation in response to the amount of nitrogen in the soil. In Lotus japonicus, two NODULE INCEPTION (NIN)-LIKE PROTEIN (NLP) transcription factors, LjNLP4 and LjNLP1, play pivotal roles in the negative regulation of nodulation by controlling the expression of symbiotic genes in high nitrate conditions. Despite an improved understanding of the molecular basis for regulating nodulation, how nitrate plays a role in the signaling pathway to negatively regulate this process is largely unknown. Here, we show that nitrate transport via NITRATE TRANSPORTER 2.1 (LjNRT2.1) is a key step in the NLP signaling pathway to control nodulation. A mutation in the LjNRT2.1 gene attenuates the nitrate-induced control of nodulation. LjNLP1 is necessary and sufficient to induce LjNRT2.1 expression, thereby regulating nitrate uptake/transport. Our data suggest that LjNRT2.1-mediated nitrate uptake/transport is required for LjNLP4 nuclear localization and induction/repression of symbiotic genes. We further show that LjNIN, a positive regulator of nodulation, counteracts the LjNLP1-dependent induction of LjNRT2.1 expression, which is linked to a reduction in nitrate uptake. These findings suggest a plant strategy in which nitrogen acquisition switches from obtaining nitrogen from the soil to symbiotic nitrogen fixation.

The nitrate transporter OsNPF7.9 mediates nitrate allocation and the divergent nitrate use efficiency between indica and japonica rice.

Nitrate allocation in Arabidopsis (Arabidopsis thaliana) represents an important mechanism for mediating plant environmental adaptation. However, whether this mechanism occurs or has any physiological/agronomic importance in the ammoniphilic plant rice (Oriza sativa L.) remains unknown. Here, we address this question through functional characterization of the Nitrate transporter 1/Peptide transporter Family (NPF) transporter gene OsNPF7.9. Ectopic expression of OsNPF7.9 in Xenopus oocytes revealed that the gene encodes a low-affinity nitrate transporter. Histochemical and in-situ hybridization assays showed that OsNPF7.9 expresses preferentially in xylem parenchyma cells of vasculature tissues. Transient expression assays indicated that OsNPF7.9 localizes to the plasma membrane. Nitrate allocation from roots to shoots was essentially decreased in osnpf7.9 mutants. Biomass, grain yield, and nitrogen use efficiency (NUE) decreased in the mutant dependent on nitrate availability. Further analysis demonstrated that nitrate allocation mediated by OsNPF7.9 is essential for balancing rice growth and stress tolerance. Moreover, our research identified an indica-japonica divergent single-nucleotide polymorphism occurring in the coding region of OsNPF7.9, which correlates with enhanced nitrate allocation to shoots of indica rice, revealing that divergent nitrate allocation might represent an important component contributing to the divergent NUE between indica and japonica subspecies and was likely selected as a favorable trait during rice breeding.

Multi-Omic Investigation of Low-Nitrogen Conditional Resistance to Clubroot Reveals Brassica napus Genes Involved in Nitrate Assimilation.

Nitrogen fertilization has been reported to influence the development of clubroot, a root disease of Brassicaceae species, caused by the obligate protist Plasmodiophora brassicae. Our previous works highlighted that low-nitrogen fertilization induced a strong reduction of clubroot symptoms in some oilseed rape genotypes. To further understand the underlying mechanisms, the response to P. brassicae infection was investigated in two genotypes “Yudal” and HD018 harboring sharply contrasted nitrogen-driven modulation of resistance toward P. brassicae. Targeted hormone and metabolic profiling, as well as RNA-seq analysis, were performed in inoculated and non-inoculated roots at 14 and 27 days post-inoculation, under high and low-nitrogen conditions. Clubroot infection triggered a large increase of SA concentration and an induction of the SA gene markers expression whatever the genotype and nitrogen conditions. Overall, metabolic profiles suggested that N-driven induction of resistance was independent of SA signaling, soluble carbohydrate and amino acid concentrations. Low-nitrogen-driven resistance in “Yudal” was associated with the transcriptional regulation of a small set of genes, among which the induction of NRT2- and NR-encoding genes. Altogether, our results indicate a possible role of nitrate transporters and auxin signaling in the crosstalk between plant nutrition and partial resistance to pathogens.