Nitrogen Assimilation Research Articles

The tetraploid wheat (Triticum dicoccum (Schrank) Schuebl.) improves nitrogen uptake and assimilation adaptation to nitrogen-deficit stress.

The genetic diversity in tetraploid wheat provides a genetic pool for improving wheat productivity and environmental resilience. The tetraploid wheat had strong N uptake, translocation, and assimilation capacity under N deficit stress, thus alleviating growth inhibition and plant N loss to maintain healthy development and adapt to environments with low N inputs. Tetraploid wheat with a rich genetic variability provides an indispensable genetic pool for improving wheat yield. Mining the physiological mechanisms of tetraploid wheat in response to nitrogen (N) deficit stress is important for low-N-tolerant wheat breeding. In this study, we selected emmer wheat (Kronos, tetraploid), Yangmai 25 (YM25, hexaploid), and Chinese spring (CS, hexaploid) as materials. We investigated the differences in the response of root morphology, leaf and root N accumulation, N uptake, translocation, and assimilation-related enzymes and gene expression in wheat seedlings of different ploidy under N deficit stress through hydroponic experiments. The tetraploid wheat (Kronos) had stronger adaptability to N deficit stress than the hexaploid wheats (YM25, CS). Kronos had better root growth under low N stress, expanding the N uptake area and enhancing N uptake to maintain higher NO3- and soluble protein contents. Kronos exhibited high TaNRT1.1, TaNRT2.1, and TaNRT2.2 expression in roots, which promoted NO3- uptake, and high TaNRT1.5 and TaNRT1.8 expression in roots and leaves enhanced NO3- translocation to the aboveground. NR and GS activity in roots and leaves of Kronos was higher by increasing the expression of TANIA2, TAGS1, and TAGS2, which enhanced the reduction and assimilation of NO3- as well as the re-assimilation of photorespiratory-released NH4+. Overall, Kronos had strong N uptake, translocation, and assimilation capacity under N deficit stress, alleviating growth inhibition and plant N loss and thus maintaining a healthy development. This study reveals the physiological mechanisms of tetraploid wheat that improve nitrogen uptake and assimilation adaptation under low N stress, which will provide indispensable germplasm resources for elite low-N-tolerant wheat improvement and breeding.

Antisense RNA regulates glutamine synthetase in a heterocyst-forming cyanobacterium.

Glutamine synthetase (GS) is a key enzyme involved in nitrogen assimilation and the maintenance of C/N balance, and it is strictly regulated in all bacteria. In cyanobacteria, glutamine synthetase expression is controlled by nitrogen control A (NtcA) transcription factor, which operates global nitrogen regulation in these photosynthetic organisms. Furthermore, post-translational regulation of GS is operated by protein-protein interaction with GS inactivating factors (IFs). Here, we describe an additional regulatory mechanism involving an antisense RNA. In Nostoc sp. PCC 7120, the GS inactivating factor A (gifA) gene (encoding GS inactivating factor IF7) is transcribed downstream of the glutamine synthetase (glnA) gene, from the opposite strand, and the gifA mRNA extends into the glnA coding sequence in antisense orientation. Therefore, the dual RNA transcript that encodes gifA constitutes two functional regions: a 5' protein-coding region, encoding IF7, and a 3' untranslated region that acts as an antisense to glnA. By increasing the levels of such antisense RNA either in cis or in trans, we demonstrate that the amount of GS activity can be modulated by the presence of the antisense RNA. The tail-to-tail disposition of the glnA and gifA genes observed in many cyanobacterial strains from the Nostocales clade suggests the prevalence of such antisense RNA-mediated regulation of GS in this group of cyanobacteria.

Different nitrogen concentrations affect strawberry seedlings nitrogen form preferences through nitrogen assimilation and metabolic pathways

The preference for different forms of nitrogen (N) is an essential means to improve plant nitrogen utilization efficiency (NUE). Currently, little information is known about the N uptake preferences of strawberries and their interaction with N concentration. In this study, the preference and distribution of nitrate nitrogen (NO3−-N) and ammonium nitrogen (NH4+-N) in strawberry seedlings were investigated in response to different N concentrations. This study used two labeled N sources (15NH4NO3 and NH415NO3) for tracer labeling. Three levels of NH4NO3 were set: low N 5 mM (N1), medium N 15 mM (N2), and high N 25 mM (N3). Each N level included two 15N labeling treatments, totaling six treatments. The results showed that under low N, medium N, and high N conditions, strawberry seedlings exhibited preferences for NH4+-N, NO3−-N, and NH4+-N, respectively. Compared with N1 and N3 treatments, N2 treatment significantly increased the activity of NR, NiR, GS, and GOGAT in strawberry seedlings roots and leaves, and the relative expression levels of FaNRT1.1, FaNRT2.1, and FaAMT1.1 in roots were significantly upregulated; When the N supply concentration increases, the distribution center of NO3−-N in strawberry seedlings shifts from stem to leaf, and under high N conditions, NH4+-N accumulates significantly in the strawberry roots. According our research, the N assimilation and metabolic capacity of strawberries themselves are important factors affecting N form preferences.

Endophytic Bacillus pumilus G5 Interacting with Silicon to Improve Drought Stress Resilience in Glycyrrhiza uralensis Fisch. by Modulating Nitrogen Absorption, Assimilation, and Metabolism Pathways.

Drought stress has become the primary severe threat to global agriculture production, including medicinal plants. Plant growth-promoting bacteria (PGPB) and environmentally friendly element silicon (Si) have emerged as effective methods in alleviating drought stress in various plants. Here, the effects of the plant endophytic G5 interaction with Si on regulating nitrogen absorption, assimilation, and metabolism pathways were investigated in the morphophysiological and gene attributes of Glycyrrhiza uralensis exposed to drought. Results showed that G5+Si application improved nitrogen absorption and assimilation by increasing the available nitrogen content in the soil, further improving the nitrogen utilization efficiency. Then, G5+Si triggered the accumulation of the major adjustment substances proline, γ-aminobutyric acid, putrescine, and chlorophyll, which played an important role in contributing to maintaining balance and energy supply in G. uralensis exposed to drought. These findings will provide new ideas for the combined application of PGPR and Si on both soil and plant systems in a drought habitat.

Size Effects of Copper Oxide Nanoparticles on Boosting Soybean Growth via Differentially Modulating Nitrogen Assimilation.

Nanoscale agrochemicals have been widely used in sustainable agriculture and may potentially affect the nitrogen fixation process in legume crops. The present study investigated the size-effects of copper oxide nanoparticles (CuO NPs) on nitrogen assimilation in soybean (G. max (L.) Merrill) plants, which were treated with different sizes (20 and 50 nm) of CuO NPs at low use doses (1 and 10 mg/kg) for 21 days under greenhouse conditions. The results showed that 50 nm CuO NPs significantly increased the fresh biomass more than 20 nm CuO NPs achieved at 10 mg/kg. The activities of N assimilation-associated enzymes and the contents of nitrogenous compounds, including nitrates, proteins, and amino acids, in soybean tissues were greatly increased across all the CuO NP treatments. The use doses of two sizes of CuO NPs had no impact on the Cu contents in shoots and roots but indeed increased the Cu contents in soils in a dose-dependent fashion. Overall, our findings demonstrated that both 20 and 50 nm CuO NPs could positively alter soybean growth and boost N assimilation, furthering our understanding that the application of nanoscale micro-nutrient-related agrochemicals at an optimal size and dose will greatly contribute to increasing the yield and quality of crops.

Systematic characterization of Gossypium GLN family genes reveals a potential function of GhGLN1.1a regulates nitrogen use efficiency in cotton

The enzyme glutamine synthetase (GLN) is mainly responsible for the assimilation and reassimilation of nitrogen (N) in higher plants. Although the GLN gene has been identified in various plants, there is little information about the GLN family in cotton (Gossypium spp.). To elucidate the roles of GLN genes in cotton, we systematically investigated and characterized the GLN gene family across four cotton species (G. raimondii, G. arboreum, G. hirsutum, and G. barbadense). Our analysis encompassed analysis of members, gene structure, cis-element, intragenomic duplication, and exploration of collinear relationships. Gene duplication analysis indicated that segmental duplication was the primary driving force for the expansion of the GhGLN gene family. Transcriptomic and quantitative real-time reverse-transcription PCR (qRT-PCR) analyses indicated that the GhGLN1.1a gene is responsive to N induction treatment and several abiotic stresses. The results of virus-induced gene silencing revealed that the accumulation and N use efficiency (NUE) of cotton were affected by the inactivation of GhGLN1.1a. This study comprehensively analyzed the GhGLN genes in Gossypium spp., and provides a new perspective on the functional roles of GhGLN1.1a in regulating NUE in cotton.

Arbuscular mycorrhizal symbiosis modulates nitrogen uptake and assimilation to enhance drought tolerance of Populus cathayana

This study aims to investigate effects of arbuscular mycorrhizal fungi (AMF) inoculation on nitrogen (N) uptake and assimilation in Populus cathayana under drought stress (DS). Herein, we measured photosynthetic performance, antioxidant enzyme system, N level and N assimilation enzymes, proteins content and distribution, transcripts of genes associated with N uptake or transport in P. cathayana with AMF (AM) or without AMF (NM) under soil water limitation and adequate irrigation. Compared with NM-DS P. cathayana, the growth, gas exchange properties, antioxidant enzyme activities, total N content and the proportion of water-soluble and membrane-bound proteins in AM-DS P. cathayana were increased. Meanwhile, nitrate reductase (NR) activity, NO3− and NO2− concentrations in AM-DS P. cathayana were reduced, while NH4+ concentration, glutamine synthetase (GS) and glutamate synthetase (GOGAT) activities were elevated, indicating that AM symbiosis reduces NO3− assimilation while promoting NH4+ assimilation. Furthermore, the transcriptional levels of NH4+ transporter genes (PcAMT1-4 and PcAMT2-1) and NO3− transporter genes (PcNRT2-1 and PcNRT3-1) in AM-DS P. cathayana roots were significantly down-regulated, as well as NH4+ transporter genes (PcAMT1-6 and PcAMT4-3) in leaves. In AM P. cathayana roots, DS significantly up-regulated the transcriptional levels of RiCPSI and RiURE, the key N transport regulatory genes in AMF compared with adequate irrigation. These results indicated that AM N transport pathway play an essential role on N uptake and utilization in AM P. cathayana to cope with DS. Therefore, this research offers a novel perspective on how AM symbiosis enhances plant resilience to drought at aspect of N acquisition and assimilation.

Impacts of AlaAT3 transgenic poplar on rhizosphere soil chemical properties, enzyme activity, bacterial community, and metabolites under two nitrogen conditions

ABSTRACT Poplar stands as one of the primary afforestation trees globally. We successfully generated transgenic poplar trees characterized by enhanced biomass under identical nutrient conditions, through the overexpression of the pivotal nitrogen assimilation gene, pxAlaAT3. An environmental risk assessment was conducted for investigate the potential changes in rhizosphere soil associated with these overexpressing lines (OL). The results show that acid phosphatase activity was significantly altered under ammonium in OL compared to the wild-type control (WT), and a similar difference was observed for protease under nitrate. 16SrDNA sequencing indicated no significant divergence in rhizosphere soil microbial community diversity between WT and OL. Metabolomics analysis revealed that the OL caused minimal alterations in the metabolites of the rhizosphere soil, posing no potential harm to the environment. With these findings in mind, we anticipate that overexpressed plants will not adversely impact the surrounding soil environment.

The transcriptomic analysis revealed the molecular mechanism of Arbuscular Mycorrhizal Fungi (AMF) inoculation in watermelon

Arbuscular mycorrhizal fungi (AMFs) and plants have a symbiotic relationship that facilitates nutrient accumulation for plant development and productivity. However, the molecular mechanism through which AMF controls the growth and development of plants is still largely unknown. In this study, the physiological and transcriptional responses of watermelon seedlings with and without AMF inoculation were compared. AMF inoculation improved the assimilation and absorption of potassium, phosphorus, and nitrogen. When inoculated, watermelon development was considerably greater than that in the absence of inoculation. Several differentially expressed genes (DEGs) were identified after 64 days after AMF inoculation. Specifically, the metabolic and signal transduction pathways exhibited an abundance of key DEGs. Inoculated watermelons resulted in 2259 DEGs involved in photosynthesis, chlorophyll biosynthesis, hormone biosynthesis, and nutrient transporters. Based on these results, it appears that AMF may enhance seedling growth and nutrient accumulation by stimulating gene expression. Notably, inoculation affected auxin-responsive proteins, auxin response factors, auxin-induced proteins, auxin-mediated proteins, and auxin transporter-like proteins, all of which are components of the auxin signalling pathway. The physiological and molecular analyses in this study may shed light on the underlying mechanism by which AMF affects the growth and development of watermelon.

Photoprotection by photoinhibitory and PSII-reaction centre quenching controls growth of Ulva rigida (Chlorophyta) and is a pre-requisite for green tide formation

Main ConclusionThe combined photoinhibitory and PSII-reaction centre quenching against light stress is an important mechanism that allows the green macroalga Ulva rigida to proliferate and form green tides in coastal ecosystems.Eutrophication of coastal ecosystems often stimulates massive and uncontrolled growth of green macroalgae, causing serious ecological problems. These green tides are frequently exposed to light intensities that can reduce their growth via the production of reactive oxygen species (ROS). To understand the physiological and biochemical mechanisms leading to the formation and maintenance of green tides, the interaction between inorganic nitrogen (Ni) and light was studied. In a bi-factorial physiological experiment simulating eutrophication under different light levels, the bloom-forming green macroalga Ulva rigida was exposed to a combination of ecologically relevant nitrate concentrations (3.8–44.7 µM) and light intensities (50–1100 µmol photons m−2 s−1) over three days. Although artificial eutrophication (≥ 21.7 µM) stimulated nitrate reductase activity, which regulated both nitrate uptake and vacuolar storage by a feedback mechanism, nitrogen assimilation remained constant. Growth was solely controlled by the light intensity because U. rigida was Ni-replete under oligotrophic conditions (3.8 µM), which requires an effective photoprotective mechanism. Fast declining Fv/Fm and non-photochemical quenching (NPQ) under excess light indicate that the combined photoinhibitory and PSII-reaction centre quenching avoided ROS production effectively. Thus, these mechanisms seem to be key to maintaining high photosynthetic activities and growth rates without producing ROS. Nevertheless, these photoprotective mechanisms allowed U. rigida to thrive under the contrasting experimental conditions with high daily growth rates (12–20%). This study helps understand the physiological mechanisms facilitating the formation and persistence of ecologically problematic green tides in coastal areas.

The Production of Bioactive Hydroxytyrosol in Fermented Beverages: The Role of Must Composition and a Genetically Modified Yeast Strain

Hydroxytyrosol (HT) is a well-known compound for its bioactive properties. It is naturally present in olives, olive oil, and wine. Its presence in wines is partly due to its production during alcoholic fermentation by yeast through a hydroxylation of tyrosol formed through the Ehrlich pathway. This work aims to explore the influence of yeast assimilable nitrogen (YAN) and glucose content as precursors of HT formation during alcoholic fermentation. Commercial Saccharomyces cerevisiae QA23 and its metabolically engineered strain were used to ferment synthetic must. Each strain was tested at two different YAN concentrations (210 and 300 mg L−1) and two glucose concentrations (100 and 240 g L−1). This work confirms that the less YAN and the more glucose, the higher the HT content, with fermentations carried out with the metabolically engineered strain being the ones with the highest HT content (0.6 mg L−1).

Nutrient depletion and heat stress impair the assimilation of nitrogen compounds in a scleractinian coral.

Concentrations of dissolved nitrogen in seawater can affect the resilience of the cnidarian-dinoflagellate symbiosis to climate change-induced bleaching. However, it is not yet known how the assimilation and translocation of the various nitrogen forms change during heat stress, nor how the symbiosis responds to nutrient depletion, which may be observed due to increasing water stratification. Here, the tropical scleractinian coral Stylophora pistillata, in symbiosis with dinoflagellates of the genus Symbiodinium, was grown at different temperatures (26°C, 30°C, and 34°C), before being placed in nutrient-replete or depleted seawater for 24 hours. The corals were then incubated with 13C-labelled sodium bicarbonate and different 15N-labelled nitrogen forms (ammonium, urea, and dissolved free amino acids) to determine their assimilation rates. We found that nutrient depletion inhibited the assimilation of all nitrogen sources studied and that heat stress reduced the assimilation of ammonium and dissolved free amino acids. However, the host assimilated over threefold more urea at 30°C relative to 26°C. Overall, both moderate heat stress (30°C) and nutrient depletion individually decreased the total nitrogen assimilated by the symbiont by 66%, and combined, they decreased assimilation by 79%. This led to the symbiotic algae becoming nitrogen starved, with the C:N ratio increasing over threefold at 34°C, potentially exacerbating the impacts of coral bleaching.

Upcycling nitrogen and curbing greenhouse gas emissions from wastewater through H2-driven assimilatory mixotrophic metabolism

Biological nitrogen assimilation is an emerging technique for the removal and upcycling of nitrogen from wastewater in the form of microbial protein (MP). For this purpose, a mixed hydrogen-oxidizing bacteria (HOB) culture capable of growing mixotrophically was evaluated for the treatment of synthetic and real wastewater under different carbon-to-nitrogen (C/N) ratios. The same mixed HOB culture was grown under heterotrophic conditions to compare treatment and process performances in terms of ammonium nitrogen (N-NH4+) removal and assimilation, chemical oxygen demand (COD) removal, CO2 release, biomass concentration, yield and protein content. Under mixotrophic conditions, the highest biomass concentration was 342.5 mg VSS∙L−1, doubling that obtained under heterotrophic conditions (161.2 mg VSS∙L−1). Discharge limits for both COD and total N were met under mixotrophic conditions, with nitrogen removal and assimilation into protein-rich biomass achieving up to 99 %. On the contrary, under heterotrophic conditions, the high content of residual nitrogen as nitrite (up to 26 mg∙L−1 of N-NO2−) did not allow to meet discharge limits for total N. The mixed HOB culture gave promising results in terms of biomass yield (0.32 g VSS·g CODH2+acetate−1) and protein content (up to 56 % of VSS) when grown mixotrophically. Under heterotrophic conditions, instead, the biomass yield (0.25 g VSS·g CODacetate−1) and protein content (35 %) were substantially lower. This study suggests that the H2-driven assimilatory mixotrophic metabolism can be successfully applied for wastewater treatment to produce effluents that meet discharge limits while mitigating greenhouse gas emissions (CO2 and N2O) and upcycling nitrogen into MP.

Effects of mycorrhizal symbiosis and Ulva lactuca seaweed extract on growth, carbon/nitrogen metabolism, and antioxidant response in cadmium-stressed sorghum plant.

In our study on the effect of cadmium (Cd) toxicity (200µM) on the growth of Sorghum bicolor (L.) Moench plants, cultivated with arbuscular mycorrhizal fungi (AMF) (Glomus intraradices) and/or under seaweed treatment (3% Ulva lactuca extract) (U. lactuca), we found that AMF increased the tolerance of sorghum to cadmium stress, either alone or in combination with the seaweed treatment. Morphological parameters were higher in these two culture conditions, with increased chlorophyll content. AMF reduced Cd accumulation in roots and inhibited its translocation to the aerial part, while seaweed treatment alone significantly increased Cd accumulation in leaves and roots without affecting plant growth compared to stressed witnesses. Treatment with AMF and/or U. lactuca attenuated oxidative stress, measured by activation of superoxide dismutase, and resulted in a significant decrease in malondialdehyde and superoxide ions (O2-) in treated plants. Furthermore, it induced significant alterations in carbon and nitrogen metabolic pathways, with a significant increase in the activity of enzymes such as glutamine synthetase, glutamate synthase (GOGAT), glutamate dehydrogenase, phosphoenolpyruvate carboxylase, aspartate aminotransferase and isocitrate dehydrogenase in the leaves of each treated plant. These results confirm that AMF, U. lactuca algae extract and their combination can improve the biochemical parameters of sorghum under Cd stress, through modification of the antioxidant response on one hand, and improved nitrogen absorption and assimilation efficiency on the other.

Resting cells of Skeletonema marinoi assimilate organic compounds and respire by dissimilatory nitrate reduction to ammonium in dark, anoxic conditions.

Diatoms can survive long periods in dark, anoxic sediments by forming resting spores or resting cells. These have been considered dormant until recently when resting cells of Skeletonema marinoi were shown to assimilate nitrate and ammonium from the ambient environment in dark, anoxic conditions. Here, we show that resting cells of S. marinoi can also perform dissimilatory nitrate reduction to ammonium (DNRA), in dark, anoxic conditions. Transmission electron microscope analyses showed that chloroplasts were compacted, and few large mitochondria had visible cristae within resting cells. Using secondary ion mass spectrometry and isotope ratio mass spectrometry combined with stable isotopic tracers, we measured assimilatory and dissimilatory processes carried out by resting cells of S. marinoi under dark, anoxic conditions. Nitrate was both respired by DNRA and assimilated into biomass by resting cells. Cells assimilated nitrogen from urea and carbon from acetate, both of which are sources of dissolved organic matter produced in sediments. Carbon and nitrogen assimilation rates corresponded to turnover rates of cellular carbon and nitrogen content ranging between 469 and 10,000 years. Hence, diatom resting cells can sustain their cells in dark, anoxic sediments by slowly assimilating and respiring substrates from the ambient environment.

A comparison of free amino nitrogen and yeast assimilable nitrogen measurement methods for use in alcoholic fermentation of whey

The continued, efficient use of whey as a fermentable substrate to produce alcoholic beverages will benefit from improving the measurement of key components relevant to the fermentability of whey. One component of fermentation that is not well studied in whey is the concentration of fermentable nitrogen, either as free amino nitrogen (FAN) or yeast assimilable nitrogen (YAN). Fermentable nitrogen in media is essential for yeast cells to replicate. Insufficient concentrations of FAN or YAN to support the growing yeast population can result in sluggish or "stuck" fermentations. Three common methods of fermentable nitrogen determination were evaluated and compared for use in whey fermentation systems, based on ninhydrin, nitrogen by o-phthalaldehyde (NOPA), and formaldehyde pH (Sørensen titration) values. Each measurement method was evaluated independently using standard addition curves and compared for their overall accuracy using paired t-tests (α = 0.05). Although the formaldehyde pH method showed high precision, it did not measure nitrogen accurately as it overestimated YAN in whey by up to 6 times relative to other tests. The ninhydrin and NOPA methods both showed accuracy for fermentable nitrogen determination in whey, based on analysis of standardized nitrogen sources. We concluded that either the ninhydrin FAN or NOPA YAN method may be used to determine the fermentable nitrogen content in whey. This is expected to improve the ability of producers to use whey as a fermentation medium.

Effects of pesticides on soil bacterial, fungal and protist communities, soil functions and grape quality in vineyards

Abstract Pesticides can have unintentional effects on non‐target organisms and change biotic communities. Such changes might be particularly important in soil microbial communities, which drive many ecosystem functions and may affect crop quality. Here, we investigated, in a three‐year study, how vegetation control (by herbicide application) and soil copper content (from long‐term copper‐based fungicide application) affect biodiversity and the community structure of soil bacteria, fungi and protists, and associated soil functions (respiration, decomposition) in Swiss vineyards. Furthermore, we determined the effects of these two management practices on grape quality as the most direct ecosystem service to farmers. Across all study years, the community composition of microorganisms was affected by herbicide application, however, a significant loss of operational taxonomic units (OTUs) was only observed in fungi and protists. Soil copper content reduced OTU richness of bacteria and protists in some years but had no significant effect on fungal richness. Copper changed the community composition in all three groups of soil microorganisms. While we found no effect of copper on soil functions, herbicide application reduced microbial respiration and biomass by about 39% and 45%, respectively. However, decomposition rates remained virtually unchanged by any pesticide. Yeast assimilable nitrogen (YAN) levels in grape were below the critical threshold of 140 mg/L in 40% of the vineyards without herbicide application and the variety Chasselas, whereas in vineyards with herbicide application, it was only 20%. Application of pesticides led to changes in richness and composition of soil microbial communities and directly reduced some soil functions (microbial biomass and respiration) but not all (decomposition). Some grape quality parameters can be indirectly enhanced by pesticide application, highlighting the trade‐off between the interests of nature conservation and the interests of the farmer. Balancing these two diverging interests requires the establishment of alternative vineyard management allowing reduced pesticide application.

Low nitrogen priming improves nitrogen uptake and assimilation adaptation to nitrogen deficit stress in wheat seedling.

Early-stage low nitrogen priming promotes root growth and delays leaf senescence through gene expression, enhancing nitrogen absorption and assimilation in wheat seedlings, thereby alleviating growth inhibition under nitrogen deficit stress and supporting normal seedling development. Verifying the strategies to reduce the amount of nitrogen (N) fertilizer while maintaining high crop yields is important for improving crop N use efficiency (NUE) and protecting the environment. To determine whether low N (LN) priming (LNP) can alleviate the impact of N-deficit stress on the growth of wheat seedlings and improve their tolerance to N-deficit stress, we conducted hydroponic experiments using two wheat cultivars, Yangmai 158 (YM158, LN tolerant) and Zaoyangmai (ZYM, LN sensitive) to study the effects of LNP on wheat seedlings under N-deficit stress. N-deficit stress decreased the plant dry weight, leaf area, and leaf N content (LNC), while LNP could significantly reduce this reduction. Distinct sensitivities to N-deficit stress were observed between the wheat cultivars, with ZYM showing an early decrease in leaf N content compared to YM158, which exhibited a late-stage reduction. LNP promoted root growth, expanded N uptake area, and upregulated the expression of TaNRT1.1, TaNRT2.1, and TaNRT2.2 in wheat seedlings, suggesting that LNP can enhance root N uptake capacity to increase N accumulation in plants. In addition, LNP improved the activity of glutamine synthase (GS) to enhance the capacity of N assimilation of plants. The relative expression of TaGS1 in the lower leaves of priming and stress (PS) was lower than that of no priming and stress (NS) after LNP, indicating that the rate of N transfer from the lower leaves to the upper leaves became slower after LNP, which alleviated the senescence of the lower leaves. The relative expression of TaGS2 was significantly increased, which might be related to the enhanced photorespiratory ammonia assimilation capacity after LNP, which reduced the N loss and maintained higher LNC. Therefore, LNP in the early stage can improve the N absorption and assimilation ability and maintain the normal N supply to alleviate the inhibition of N-deficit stress in wheat seedlings.

Environment-specific virocell metabolic reprogramming.

Viruses impact microbial systems through killing hosts, horizontal gene transfer, and altering cellular metabolism, consequently impacting nutrient cycles. A virus-infected cell, a "virocell", is distinct from its uninfected sister cell as the virus commandeers cellular machinery to produce viruses rather than replicate cells. Problematically, virocell responses to the nutrient-limited conditions that abound in nature are poorly understood. Here we used a systems biology approach to investigate virocell metabolic reprogramming under nutrient limitation. Using transcriptomics, proteomics, lipidomics, and endo- and exo-metabolomics, we assessed how low phosphate (low-P) conditions impacted virocells of a marine Pseudoalteromonas host when independently infected by two unrelated phages (HP1 and HS2). With the combined stresses of infection and nutrient limitation, a set of nested responses were observed. First, low-P imposed common cellular responses on all cells (virocells and uninfected cells), including activating the canonical P-stress response, and decreasing transcription, translation, and extracellular organic matter consumption. Second, low-P imposed infection-specific responses (for both virocells), including enhancing nitrogen assimilation and fatty acid degradation, and decreasing extracellular lipid relative abundance. Third, low-P suggested virocell-specific strategies. Specifically, HS2-virocells regulated gene expression by increasing transcription and ribosomal protein production, whereas HP1-virocells accumulated host proteins, decreased extracellular peptide relative abundance, and invested in broader energy and resource acquisition. These results suggest that although environmental conditions shape metabolism in common ways regardless of infection, virocell-specific strategies exist to support viral replication during nutrient limitation, and a framework now exists for identifying metabolic strategies of nutrient-limited virocells in nature.

Foliar urea applications to apple trees increase yeast assimilable nitrogen, amino acids, and flavor volatiles in fruit and hard cider

Volatile compounds are a crucial component of hard apple (Malus ✕domestica Borkh.) cider aroma, flavor, and consequent marketability. Amino acids are used by Saccharomyces cerevisiae yeast in the production of volatiles and their precursors. This study hypothesized that foliar applications of nitrogen to apple trees would elevate the concentration of amino acids in the fruit, thus increasing the concentration of sensorially distinguishable aromatic volatiles in cider. In this experiment, urea foliar fertilizer was applied to ‘Ellis Bitter’ and ‘Harry Masters Jersey’ apple trees to create Control (0 applications), Low (3 weekly applications), and High (5 weekly applications) treatments. Total leaf nitrogen increased with the number of foliar urea applications, demonstrating plant uptake. The total fruit yield and fruit efficiency (total harvested fruit mass relative to trunk cross-sectional area) did not differ among treatments within the same cultivar. Similarly, fruit maturity, size, total phenolics, pH, and soluble solids concentration did not differ among treatments. Yeast assimilable nitrogen (YAN) and amino acids in the juice increased with the number of foliar urea applications, as did fermentation rates. Relative to the Control, the High treatment had 130% greater YAN in ‘Ellis Bitter’ and 145% in ‘Harry Masters Jersey’. Over 90% of the YAN in all juice samples was composed of primary amino nitrogen (PAN), and the majority of the PAN among all treatments was asparagine. Ester and fatty acid content in hard ciders increased with the number of urea applications; however, higher alcohols showed a mixed relationship with the number of applications in the cultivar ‘Ellis Bitter’. Aroma and flavor differences among treatments were discernible by triangle discrimination test panelists; however, the specific sensory differences were not easily identifiable with untrained panelists. This study demonstrated that late season foliar urea applications to apples trees can increase juice YAN and aromatic compounds in finished cider.