Nitrogen Isotope Discrepancy Between Primary Producers and Sediments in an Anoxic and Alkaline Lake

Nitrogen isotope compositions (δ15N) in sedimentary rocks are extensively used to investigate the biogeochemical nitrogen cycle through geological times. This use relies on the observation that, in modern continental platforms and anoxic basins, surface sediments faithfully record the δ15N of primary producers, assuming that it was similar in the past. Over Earth’s history, however, surface environments experienced profound changes, including the transition of ammonium-dominated to nitrate-dominated waters and the transition from exclusively microbial ecosystems to ecosystems including multicellularity, which make modern environments significantly different compared to earlier ones, potentially invalidating the fundamental assumption that surface sediments faithfully record the δ15N of primary producers. In order to improve our understanding of the nitrogen isotopic information contained in the early Earth’s sedimentary rock record, we investigate here the nitrogen isotope systematics in a microbial, nitrate free and ammonium-rich modern system, the Dziani Dzaha Lake. In this modern system, the δ15N of the reduced dissolved inorganic nitrogen (i.e., NH4+ and NH3) in the water column is close to ∼7‰ . δ15N of suspended particulate matter (SPM) show a similar average value in surface waters (i.e., where SPM is massively composed of active primary producers), but increases up to 14‰ in the deeper part of the water column during periods when it is enriched in dissolved reduced species (i.e., CH4, H2S/HS− and NH4+/NH3). Surface sediments δ15N, with values comprised between 10 and 14 ‰, seem to preferentially record these positive isotopic signatures, rather than those of active primary producers. We propose here that the observed isotopic pattern is mainly linked to the assimilation of ammonium strongly enriched in 15N by isotope exchange with ammonia under basic conditions. Although ammonium assimilation seems here to be responsible for a significant isotopic enrichment due to the basic conditions, in neutral anoxic environments inhabited by similar microbial ecosystems, this process may also significantly impact the δ15N of primary producers towards more negative values. This would have strong implications for our interpretation of the Precambrian sedimentary record as this finding challenges one the fundamental hypotheses underlying the use of sedimentary δ15N in paleo-oceanographic reconstructions, i.e. that surface sediments faithfully record the δ15N of active primary producers in the photic zone.

Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in Pyropia haitanensis

Pyropia haitanensis is an important laver species in China. Its quality traits are closely related to the content of glutamic acid. Glutamate dehydrogenase (GDH) is a crucial enzyme in the glutamic acid metabolism. In this study, two GDH genes from P. haitanensis, PhGDH1 and PhGDH2, were cloned and successfully expressed in Escherichia coli. The in vitro enzyme activity assay demonstrated that the catalytic activity of PhGDHs is mainly in the direction of ammonium assimilation. The measured Km values of PhGDH1 for NADH, (NH4)2SO4, and α-oxoglutarate were 0.12, 4.99, and 0.16 mM, respectively, while the corresponding Km values of PhGDH2 were 0.02, 3.98, and 0.104 mM, respectively. Site-directed mutagenesis results showed that Gly193 and Thr361 were important catalytic residues for PhGDH2. Moreover, expression levels of both PhGDHs were significantly increased under abiotic stresses. These results suggest that PhGDHs can convert α-oxoglutarate to glutamic acid, and enhance the flavor and stress resistance of P. haitanensis.

Drought, Salinity, and Low Nitrogen Differentially Affect the Growth and Nitrogen Metabolism of Sophora japonica (L.) in a Semi-Hydroponic Phenotyping Platform

Abiotic stresses, such as salinity, drought, and nutrient deficiency adversely affect nitrogen (N) uptake and assimilation in plants. However, the regulation of N metabolism and N pathway genes in Sophora japonica under abiotic stresses is unclear. Sophora japonica seedlings were subjected to drought (5% polyethylene glycol 6,000), salinity (75mM NaCl), or low N (0.01mM NH4NO3) for 3weeks in a semi-hydroponic phenotyping platform. Salinity and low N negatively affected plant growth, while drought promoted root growth and inhibited aboveground growth. The NH4+/NO3− ratio increased under all three treatments with the exception of a reduction in leaves under salinity. Drought significantly increased leaf NO2− concentrations. Nitrate reductase (NR) activity was unaltered or increased under stresses with the exception of a reduction in leaves under salinity. Drought enhanced ammonium assimilation with increased glutamate synthase (GOGAT) activity, although glutamine synthetase (GS) activity remained unchanged, whereas salinity and low N inhibited ammonium assimilation with decreased GS activity under salt stress and decreased GOGAT activity under low N treatment. Glutamate dehydrogenase (GDH) activity also changed dramatically under different stresses. Additionally, expression changes of genes involved in N reduction and assimilation were generally consistent with related enzyme activities. In roots, ammonium transporters, especially SjAMT1.1 and SjAMT2.1a, showed higher transcription under all three stresses; however, most nitrate transporters (NRTs) were upregulated under salinity but unchanged under drought. SjNRT2.4, SjNRT2.5, and SjNRT3.1 were highly induced by low N. These results indicate that N uptake and metabolism processes respond differently to drought, salinity, and low N conditions in S. japonica seedlings, possibly playing key roles in plant resistance to environmental stress.

The Coupling of Carbon and Nitrogen in Response to Variable NH4+/ NO3- Ratios in Brassica Napus Plantlets in Vitro

BackgroundPlantlets grown in vitro with a mixed nitrogen source utilize sucrose and CO2 as carbon sources for growth. However, it is very difficult to obtain the correct proportion of assimilated nitrate, ammonium, sucrose and CO2 for plantlets. Consequently, the NH4+/NO3- use efficiency for carbon fixation derived from the assimilation of sucrose/CO2 is still unclear for plantlets. ResultsThe bidirectional stable nitrogen isotope tracer technique was employed to quantify the proportions of nitrate and ammonium utilized at different NH4+/ NO3- ratios, and the proportions of sucrose and CO2 assimilation were quantified by the foliar δ13C values of plantlets. There was an obvious difference in the assimilation of nitrate and ammonium under different NH4+/NO3- ratios for Brassica napus (Bn) plantlets. Increasing the supply of nitrate contributed to enhancing the assimilation of nitrate and ammonium simultaneously. The nitrate utilization coefficients of the Bn plantlets had no distinct change with increasing nitrate concentration, while the ammonium utilization coefficients of the Bn plantlets increased obviously with increasing nitrate concentration. The proportion of sucrose/CO2 assimilation depended on the NH4+/NO3- ratios of the Bn plantlets. Both nitrate and ammonium assimilation were independent of sucrose/CO2 assimilation. Based on the proportion of CO2, sucrose, nitrate and ammonium utilization, the nitrate/ammonium use efficiency (as indicated by the C/N ratio) for carbon fixation derived from the assimilation of sucrose/CO2 can be quantified for Bn plantlets.ConclusionsQuantifying the utilization proportions of nitrate and ammonium can reveal the difference in nitrate and ammonium utilization among plantlets at different NH4+/NO3- ratios. Foliar δ13C value in combination of the foliar δ15N value of plantlets can be used to quantify the nitrate/ammonium use efficiency for the carbon fixation derived from the assimilation of sucrose/CO2, which contributes to knowing the coupling process of carbon and nitrogen in plantlets and provides an alternate way to optimize the supply of inorganic nitrogen in culture media.

Impacts of environmental conditions, and allelic variation of cytosolic glutamine synthetase on maize hybrid kernel production

Cytosolic glutamine synthetase (GS1) is the enzyme mainly responsible of ammonium assimilation and reassimilation in maize leaves. The agronomic potential of GS1 in maize kernel production was investigated by examining the impact of an overexpression of the enzyme in the leaf cells. Transgenic hybrids exhibiting a three-fold increase in leaf GS activity were produced and characterized using plants grown in the field. Several independent hybrids overexpressing Gln1-3, a gene encoding cytosolic (GS1), in the leaf and bundle sheath mesophyll cells were grown over five years in different locations. On average, a 3.8% increase in kernel yield was obtained in the transgenic hybrids compared to controls. However, we observed that such an increase was simultaneously dependent upon both the environmental conditions and the transgenic event for a given field trial. Although variable from one environment to another, significant associations were also found between two GS1 genes (Gln1-3 and Gln1-4) polymorphic regions and kernel yield in different locations. We propose that the GS1 enzyme is a potential lead for producing high yielding maize hybrids using either genetic engineering or marker-assisted selection. However, for these hybrids, yield increases will be largely dependent upon the environmental conditions used to grow the plants.

Redox-Controlled Ammonium Storage and Overturn in Ediacaran Oceans

As a key nutrient, nitrogen can limit primary productivity and carbon cycle dynamics, but also evolutionary progress. Given strong redox-dependency of its molecular speciation, environmental conditions can control nitrogen localization and bioavailability. This particularly applies to periods in Earth history with strong and frequent redox fluctuations, such as the Neoproterozoic. We here report on chlorophyll-derived porphyrins and maleimides in Ediacaran sediments from Oman. Exceptionally light δ15N values (< –10‰) in maleimides derived from anoxygenic phototrophs point towards ammonium assimilation at the chemocline, whereas the isotopic offset between kerogens and chlorophyll-derivatives indicates a variable regime of cyanobacterial and eukaryotic primary production in surface waters. Biomarker and maleimide mass balance considerations imply shallow euxinia during the terminal Ediacaran and a stronger contribution of anoxygenic phototrophs to primary productivity, possibly as a consequence of nutrient ‘lockup’ in a large anoxic ammonium reservoir. Synchronous δ13C and δ15N anomalies at the Ediacaran–Cambrian boundary may reflect one in a series of overturn events, mixing ammonium and isotopically-light DIC into oxic surface waters. By modulating access to nitrogen, environmental redox conditions may have periodically affected Ediacaran primary productivity, carbon cycle perturbations, and possibly played a role in the timing of the metazoan radiation across the terminal Ediacaran and early Cambrian.

MdUGT88F1-mediated phloridzin biosynthesis coordinates carbon and nitrogen accumulation in apple

The high accumulation of phloridzin makes apple (Malus domestica) unique in the plant kingdom, which suggests a vital role of its biosynthesis in the physiological processes of apple. In our previous study, silencing MdUGT88F1 (a key UDP-glucose: phloretin 2'-O-glucosyltransferase gene) revealed the importance of phloridzin biosynthesis in apple development and Valsa canker resistance. Here, results from MdUGT88F1-silencing lines showed that phloridzin biosynthesis was indispensable for normal chloroplast development and photosynthetic carbon fixation by maintaining MdGLK1/2 expression. Interestingly, the increased phloridzin biosynthesis didn’t affect plant (or chloroplast) development but reduced nitrogen accumulation, leading to chlorophyll deficiency, light sensitivity, and sugar accumulation in MdUGT88F1-overexpressing apple lines during their growth and development. Further analysis revealed that MdUGT88F1-mediated phloridzin biosynthesis negatively regulated cytosolic glutamine synthetase1-asparagine synthetase-asparaginase (GS1-AS-ASPG) pathway of ammonium assimilation and limited chlorophyll synthesis in the shoots of apple. The interference of phloridzin biosynthesis in the GS1-AS-ASPG pathway was also assumed to be associated with its limitation of the carbon skeletons of ammonium assimilation through metabolic competition with the tricarboxylic acid cycle. Taken together, our findings shed light on the role of MdUGT88F1-mediated phloridzin biosynthesis in the coordination between carbon and nitrogen accumulation in apple trees.

Systemic regulation of nitrogen acquisition and use in Oryza longistaminata ramets under nitrogen heterogeneity

Oryza longistaminata, a wild rice, vegetatively reproduces and forms a networked clonal colony consisting of ramets connected by rhizomes. Although water, nutrients, and other molecules can be transferred between ramets via the rhizomes, inter-ramet communication in response to spatially heterogeneous nitrogen availability is not well understood. We studied the response of ramet pairs to heterogeneous nitrogen availability by using a split hydroponic system that allowed each ramet root to be exposed to different conditions. Ammonium uptake was compensatively enhanced in the sufficient-side root when roots of the ramet pairs were exposed to ammonium-sufficient and deficient conditions. Comparative transcriptome analysis revealed that a gene regulatory network for effective ammonium assimilation and amino acid biosynthesis was activated in the sufficient-side roots. Allocation of absorbed nitrogen from the nitrogen-sufficient to the deficient ramets was rather limited. Nitrogen was preferentially used for newly growing axillary buds on the sufficient-side ramets. Biosynthesis of trans-zeatin, a cytokinin, was up-regulated in response to the nitrogen supply, but trans-zeatin appears not to target the compensatory regulation. Our results also implied that the O. longistaminata ortholog of OsCEP1 plays a role as a nitrogen-deficient signal in inter-ramet communication, providing compensatory up-regulation of nitrogen assimilatory genes. These results provide insights into the molecular basis for efficient growth strategies of asexually proliferating plants growing in areas where nitrogen distribution is spatially heterogeneous.