Production and cross-feeding of nitrite within Prochlorococcus populations

Prochlorococcus is an abundant photosynthetic bacterium in the oligotrophic open ocean where nitrogen (N) often limits the growth of phytoplankton. Prochlorococcus has evolved into multiple phylogenetic clades of high-light (HL) adapted and low-light (LL) adapted cells. Within these clades, cells encode a variety of N assimilation traits that are differentially distributed among members of the population. Among these traits, nitrate (NO3-) assimilation is generally restricted to a few clades of high-light adapted cells (the HLI, HLII, and HLVI clades) and a single clade of low-light adapted cells (the LLI clade). Most, if not all, cells belonging to the LLI clade have the ability to assimilate nitrite (NO2-), with a subset of this clade capable of assimilating both NO3- and NO2-. Cells belonging to the LLI clade are maximally abundant at the top of the nitracline and near the primary NO2- maximum layer. In some ecosystems, this peak in NO2- concentration may be a consequence of incomplete assimilatory NO3- reduction by phytoplankton. This phenomenon is characterized by a bottleneck in the downstream half of the NO3- assimilation pathway and the concomitant accumulation and release of NO2- by phytoplankton cells. Given the association between LLI Prochlorococcus and the primary NO2- maximum layer, we hypothesized that some Prochlorococcus exhibit incomplete assimilatory NO3- reduction. To assess this, we monitored NO2- accumulation in batch culture for 3 Prochlorococcus strains (MIT0915, MIT0917, and SB) and 2 Synechococcus strains (WH8102 and WH7803) when grown on NO3- as the sole N source. Only MIT0917 and SB accumulated external NO2- during growth on NO3-. Approximately 20-30% of the NO3- transported into the cell by MIT0917 was released as NO2-, with the balance assimilated into biomass. We further observed that co-cultures using NO3- as the sole N source could be established for MIT0917 and a Prochlorococcus strain that can assimilate NO2- but not NO3-. In these co-cultures, the NO2- released by MIT0917 was efficiently consumed by its partner strain during balanced exponential growth. Our findings highlight the potential for emergent metabolic partnerships within Prochlorococcus populations that are mediated by the production and consumption of the N cycle intermediate, NO2-.

Foliar Application of Protein Hydrolysates on Baby-Leaf Spinach Grown at Different N Levels

Surpluses of N are associated with environmental and health problems. To optimise N use and reduce nitrate accumulation in leafy species like spinach, the application of biostimulants is suggested. An experiment in controlled conditions (growth chamber/soilless) evaluated baby-spinach responses to two protein hydrolysates (PHs) from plant (legume, Trainer®) and animal (meat, Isabion®) sources, combined with three N rates: 2 (N2, deficient), 8 (N8, sub-optimal), and 14 (N14, optimal) mM of N. Biometrical and morphological traits of shoots and roots as well as the physio-metabolic (gas exchange, N assimilation, and NUtE), physical, mineral, and antioxidant profiles of leaves were assessed. The legume-PH boosts growth and yield only at the highest N conditions, while there was no effect at lower N rates. The legume-PH modulates root architecture and chlorophylls has positive responses only at optimal N availability, such as an increase in N uptake, leaf expansion, and photosynthetic activity at the canopy level. The PHs do not improve NUtE, leaf colour, consistency, cations, or antioxidants. Neither do PHs have any effect on reducing nitrate accumulation. Legume-PH improves N assimilation only at optimal N availability, while meat-PH does not, reaching the highest nitrate value at the highest N rate (2677 mg kg−1 fw), even if this value is under the EC limits for fresh spinach.

Growth, Quality, and Nitrogen Assimilation in Response to High Ammonium or Nitrate Supply in Cabbage (Brassica campestris L.) and Lettuce (Lactuca sativa L.)

Plants grow better when they are supplied with a combination of ammonium (NH4+) and nitrate (NO3−) than when either one is supplied as the sole N (nitrogen) source. However, the effects of N forms on N metabolism and major N assimilation enzymes in different plants, especially vegetables, are largely neglected. This study was conducted on two plants with distinct NH4+ tolerances to compare the responses of two popular leafy vegetables, Korean cabbage (Brassica campestris L.) ‘Ssamchu’ and lettuce (Lactuca sativa L.) ‘Caesar green’, to the N source. To this end, plant growth and quality, photosynthesis, carbohydrate, N contents (in the forms of NO3−, NO2−, NH4+, total protein), and key N assimilation-related enzyme (NR, NIR, GS, GDH) activities were investigated. When plants were subjected to one of three NH4+:NO3− regimes, 0:100, 50:50, or 100:0, lettuce was relatively more tolerant while cabbage was extremely sensitive to high NH4+. Both plants benefited more from being grown with 50:50 NH4+:NO3−, as evidenced by the best growth performance, ameliorated photosynthesis, and enriched carbohydrate (C) stock content. In addition, as compared to cabbage, the GS and GDH activities were reinforced in lettuce in response to an increasing external NH4+ level, resulting in low NH4+ accumulation. Our findings suggested that boosting or maintaining high GS and GDH activities is an important strategy for the ammonium tolerance in vegetables.

Comparing the Effects of N and P Deficiency on Physiology and Growth for Fast- and Slow-Growing Provenances of Fraxinus mandshurica

With the continuous increase in atmospheric carbon dioxide emissions, nitrogen (N) and phosphorus (P) as mineral elements increasingly restrict plant growth. To explore the effect of deficiency of P and N on growth and physiology, Fraxinus mandshurica (hereafter “F. mandshurica”) Rupr. annual seedlings of Wuchang (WC) provenance with fast growth and Dailing (DL) provenance with slow growth were treated with complete nutrition or starvation of N (N-), P (P-) or both elements (NP-). Although P- and N- increased the use efficiency of P (PUE) and N (NUE), respectively, they reduced the leaf area, chlorophyll content and activities of N assimilation enzymes (NR, GS, GOGAT), which decreased the dry weight and P or N amount. The free amino acid content and activities of Phosphoenolpyruvate carboxylase (PEPC) and acid phosphatase enzymes were reduced by N-. The transcript levels of NRT2.1, NRT2.4, NRT2.5, NRT2.7, AVT1, AAP3, NIA2, PHT1-3, PHT1-4 and PHT2-1 in roots were increased, but those of NRT2.1, NRT2.4, NRT2.5, PHT1-3, PHT1-4, PHT2-1 and AAP3 in leaves were reduced by P-. WC was significantly greater than DL under P- in dry weight, C amount, N amount, leaf area, PUE, NUE, which related to greater chlorophyll content, PEPC enzyme activity, N assimilation enzyme activities, and transcript levels of N and P transporter genes in roots and foliage, indicating a greater ability of WC to absorb, transport and utilize N and P under P-. WC was also greater than DL under N- in terms of the above indicators except the transcript levels of N and P assimilation genes, but most of the indicators did not reach a significant level, indicating that WC might be more tolerant to N- than DL, which requires further verification. In summary, WC was identified as a P-efficient provenance, as the growth rate was greater for the genetic type with high than low tolerance to P-.

A mechanistic investigation of enhanced nitrogen use efficiency in wheat seedlings after treatment with an Ascophyllum nodosum biostimulant

ABSTRACTReduction in the emissions of the greenhouse gas nitrous oxide and nitrogen (N) pollution of ground water by improving nitrogen use efficiency (NUE) in crops is urgently required in pursuit of a sustainable agricultural future. Utilising an engineered biostimulant (PSI-362) derived from the brown seaweed Ascophyllum nodosum, we examined its effect on wheat seedling growth dynamics and mechanistic spatiotemporal changes at transcriptional and biochemical levels in relation to N uptake, assimilation and NUE. PSI-362-mediated biomass increase was associated with increased nitrate uptake and N assimilation in the form of glutamate, glutamine, free amino acids, soluble proteins and total 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), and assimilation (TaNR1 and TaNiR1, TaGDH2, TaGoGAT, TaGS1). Finally, a comparative analysis of the PSI-362 and two generic Ascophyllum nodosum extracts (ANEs) demonstrated that the NUE effect greatly differs depending on the ANE biostimulant used. In the current context of climate warming the transition of agriculture to a more sustainable model is urgently required. Application and adoption of precision biostimulants creates an opportunity for sustainable crop management, reduced production cost and environmental pollution, while maintaining yields.

Nitrogen deficiency regulates premature senescence by modulating flag leaf function, ROS homeostasis, and intercellular sugar concentration in rice during grain filling

Background Leaf senescence occurs in an age-dependent manner, but the rate and timing of leaf senescence may be influenced by various biotic and abiotic factors. In the course of stress, the function, composition, and different components of photosynthetic apparatus occur to be synthesized homogeneously or degraded paradoxically due to different senescence-related processes. Nitrogen (N) deficiency is one of the critical environmental factors that induce leaf senescence, and its incidence may curtail leaf photosynthetic function and markedly alter the genetic information of plants that might result in low grain yield. However, the physiological and genetic mechanism underlying N deficiency regulates premature senescence, and flag leaf function, ROS homeostasis, and intercellular sugar concentration in rice during grain filling are not well understood. In this paper, Zhehui7954 an excellent indica restorer line (wildtype) and its corresponding mutant (psf) with the premature senescence of flag leaves were used to study the effect of different N supplies in the alteration of physiological and biochemical components of flag leaf organ and its functions during grain filling. Results The results showed that the psf mutant appeared to be more susceptible to the varying N supply levels than WT. For instance, the psf mutant showed considerably lower Pn, Chl a, Chl b, and Car contents than its WT. N deficiency (LN) decreased leaves photosynthetic activities, N metabolites, but significantly burst O2•−, H2O2, and relative conductivity (R1/R2) concentrations, which was consistent with the expression levels of senescence-associated genes. Sucrose, glucose, and C/N ratio concentrations increased with a decrease in N level, which was closely associated with N and non-structural carbohydrate translocation rates. Increases in POD activity were positively linked with the senescence-related enhancement of ROS generation under LN conditions, whereas, SOD, CAT, and APX activities showed opposite trends. High N (HN) supply significantly inhibits the transcripts of carbohydrate biosynthesis genes, while N assimilation gene transcripts gradually increased along with leaf senescence. The psf mutant had a relatively higher grain yield under HN treatment than LN, while WT had a higher grain yield under MN than HN and LN. Conclusions This work revealed that the C/N ratio and ROS undergo a gradual increase driven by interlinking positive feedback, providing a physiological framework connecting the participation of sugars and N assimilation in the regulation of leaf senescence. These results could be useful for achieving a higher yield of rice production by appropriate N supply and plant senescence regulation.

Integrative transcriptomic and metabolomic analysis of D-leaf of seven pineapple varieties differing in N-P-K% contents

Background Pineapple (Ananas comosus L. Merr.) is the third most important tropical fruit in China. In other crops, farmers can easily judge the nutritional requirements from leaf color. However, concerning pineapple, it is difficult due to the variation in leaf color of the cultivated pineapple varieties. A detailed understanding of the mechanisms of nutrient transport, accumulation, and assimilation was targeted in this study. We explored the D-leaf nitrogen (N), phosphorus (P), and potassium (K) contents, transcriptome, and metabolome of seven pineapple varieties. Results Significantly higher N, P, and K% contents were observed in Bali, Caine, and Golden pineapple. The transcriptome sequencing of 21 libraries resulted in the identification of 14,310 differentially expressed genes in the D-leaves of seven pineapple varieties. Genes associated with N transport and assimilation in D-leaves of pineapple was possibly regulated by nitrate and ammonium transporters, and glutamate dehydrogenases play roles in N assimilation in arginine biosynthesis pathways. Photosynthesis and photosynthesis-antenna proteins pathways were also significantly regulated between the studied genotypes. Phosphate transporters and mitochondrial phosphate transporters were differentially regulated regarding inorganic P transport. WRKY, MYB, and bHLH transcription factors were possibly regulating the phosphate transporters. The observed varying contents of K% in the D-leaves was associated to the regulation of K+ transporters and channels under the influence of Ca2+ signaling. The UPLC-MS/MS analysis detected 873 metabolites which were mainly classified as flavonoids, lipids, and phenolic acids. Conclusions These findings provide a detailed insight into the N, P, K% contents in pineapple D-leaf and their transcriptomic and metabolomic signatures.

Assimilation of Particular Organic Matter and Dissolved Organic or Inorganic Compounds by Cribroelphidium selseyense (Foraminifera)

Marine carbon and nitrogen processing through microorganisms’ metabolism is an important aspect of the global element cycles. For that purpose, we used foraminifera to analyze the element turnover with different algae food sources. In the Baltic Sea, benthic foraminifera are quite common and therefore it is important to understand their metabolism. Especially, Cribroelphidium selseyense, also occurring in the Baltic Sea, has often been used for laboratory feeding experiments to test their effect on carbon or nitrogen turnover. Therefore, foraminifera were collected from the Kiel Fjord and fed with six different algal species in two qualities (freeze-dried algae vs. fresh algae, all 13C- and 15N-labeled). Also, labeled dissolved inorganic C and N compounds and glucose were offered to the foraminifera to test direct assimilation of dissolved compounds (carbon and nitrogen) from the water column. Our experiments showed that after 15 days of incubation, there were highly significant differences in isotope labeling in foraminifera fed with fresh algae and dry algae, depending on algal species. Further, different algal species led to different 13C and 15N enrichment in the studied foraminifera, highlighting a feeding preference for one diatom species and an Eustigmatophyte. A significant carbon assimilation from HCO3– was observed after 7 days of incubation. The N assimilation from NH4+ was significantly higher than for NO3– as an inorganic N source. The uptake of glucose showed a lag phase, which was often observed during past experiments, where foraminifera were in a steady state and showed no food uptake at regular intervals. These results highlight the importance of food quality on the feeding behavior and metabolic pathways for further studies of foraminiferal nutrition and nutrient cycling.

Nitrogen management under increased atmospheric CO2 concentration in cucumber (Cucumis sativus L.): ameliorating environmental impacts of fertilization

In the last years, the atmospheric CO2 concentration has increased significantly, and this increase can cause changes in various physiological and biochemical processes of plants. However, the response of plants to elevated CO2 concentration (e[CO2]) will be different depending on the nitrogen form available and the plant species. Therefore, hydroponic trials on cucumber plants, with two CO2 concentrations (400 and 1000 ppm) and two nitrogen sources (NO3−/NH4+; 100/0 and 90/10), were conducted. Physiological parameters—such as gas exchange, GS, GOGAT and GDH activities, cation composition, soluble sugar and starch content- were measured. The results showed that when plants were grown with NH4+ and e[CO2], parameters such as photosynthesis rate (ACO2), instantaneous water use efficiency (WUEi), the content of NH4+, Ca2+ and Mg2+, and the concentration of starch, were higher than in control plants (irrigated with nitrate as sole nitrogen source and ambient CO2 concentration (a[CO2])). Furthermore, an improvement in N assimilation was observed when the GS/GOGAT pathway was enhanced under these conditions (NH4+ and e[CO2]). Thus, our results contribute to the reduction of the negative environmental impacts of the use of nitrogen fertilizers on this crop, both by reducing nitrogen leakage (eutrophication) and greenhouse gas emissions.