Certain Environmental Conditions Maximize Ammonium Accumulation and Minimize Nitrogen Loss During Nitrate Reduction Process by Pseudomonas putida Y-9

Realizing the smallest nitrogen loss is a challenge in the nitrate reduction process. Dissimilatory nitrate reduction to ammonium (DNRA) and nitrate assimilation play crucial roles in nitrogen retention. In this study, the effects of the carbon source, C/N ratio, pH, and dissolved oxygen on the multiple nitrate reduction pathways conducted by Pseudomonas putida Y-9 are explored. Strain Y-9 efficiently removed nitrate (up to 89.79%) with glucose as the sole carbon source, and the nitrogen loss in this system was 15.43%. The total nitrogen decrease and ammonium accumulation at a C/N ratio of 9 were lower than that at 12 and higher than that at 15, respectively (P < 0.05). Besides, neutral and alkaline conditions (pH 7–9) favored nitrate reduction. Largest nitrate removal (81.78%) and minimum nitrogen loss (10.63%) were observed at pH 7. The nitrate removal and ammonium production efficiencies of strain Y-9 increased due to an increased shaking speed. The expression patterns of nirBD (the gene that controls nitrate assimilation and DNRA) in strain Y-9 were similar to ammonium patterns of the tested incubation conditions. In summary, the following conditions facilitated nitrate assimilation and DNRA by strain Y-9, while reducing the denitrification: glucose as the carbon source, a C/N ratio of 9, a pH of 7, and a shaking speed of 150 rpm. Under these conditions, nitrate removal was substantial, and nitrogen loss from the system was minimal.

Excessive ammonium assimilation by plastidic glutamine synthetase causes ammonium toxicity in Arabidopsis thaliana

Plants use nitrate, ammonium, and organic nitrogen in the soil as nitrogen sources. Since the elevated CO2 environment predicted for the near future will reduce nitrate utilization by C3 species, ammonium is attracting great interest. However, abundant ammonium nutrition impairs growth, i.e., ammonium toxicity, the primary cause of which remains to be determined. Here, we show that ammonium assimilation by GLUTAMINE SYNTHETASE 2 (GLN2) localized in the plastid rather than ammonium accumulation is a primary cause for toxicity, which challenges the textbook knowledge. With exposure to toxic levels of ammonium, the shoot GLN2 reaction produced an abundance of protons within cells, thereby elevating shoot acidity and stimulating expression of acidic stress-responsive genes. Application of an alkaline ammonia solution to the ammonium medium efficiently alleviated the ammonium toxicity with a concomitant reduction in shoot acidity. Consequently, we conclude that a primary cause of ammonium toxicity is acidic stress.

Biogeochemical controls on wintertime ammonium accumulation in the surface layer of the Southern Ocean

The production and consumption of ammonium (NH4+) are essential upper-ocean nitrogen cycle pathways, yet in the Southern Ocean where NH4+ has been observed to accumulate in surface waters, its mixed-layer cycling remains poorly understood. For surface samples collected between Cape Town and the marginal ice zone (MIZ) in winter 2017, we found that NH4+ concentrations were five-fold higher than is typical for summer, and lower north than south of the Subantarctic Front (SAF; 0.01–0.26 µM versus 0.19–0.70 µM). Our observations confirm that NH4+ accumulates in the Southern Ocean’s winter mixed layer, particularly in polar waters. NH4+ uptake rates were highest near the Polar Front (PF; 12.9 ± 0.4 nM day−1) and in the Subantarctic Zone (10.0 ± 1.5 nM day−1), decreasing towards the MIZ (3.0 ± 0.8 nM day−1) despite high ambient NH4+ concentrations, likely due to low sea surface temperatures and light availability. By contrast, rates of NH4+ oxidation were higher south than north of the PF (16.0 ± 0.8 versus 11.1 ± 0.5 nM day−1), perhaps due to the lower light and higher iron conditions characteristic of polar waters. Augmenting our dataset with NH4+ concentration measurements spanning the 2018/2019 annual cycle reveals that mixed-layer NH4+ accumulation south of the SAF likely derives from sustained heterotrophic NH4+ production in late summer through winter that outpaces NH4+ consumption by temperature-, light, and iron-limited microorganisms. Our observations thus imply that the Southern Ocean becomes a biological source of CO2 to the atmosphere for half the year not only because nitrate drawdown is weak, but also because the ambient conditions favour net heterotrophy and NH4+ accumulation.

Seasonal Stratification and Biogeochemical Turnover in the Freshwater Reach of a Partially Mixed Dredged Estuary

The Elbe estuary is a substantially engineered tidal water body that receives high loads of organic matter from the eutrophied Elbe river. The organic matter entering the estuary at the tidal weir is dominated by diatom populations that collapse in the deepened freshwater reach. Although the estuary’s freshwater reach is considered to manifest vertically homogenous density distribution (i.e., to be well-mixed), several indicators like trapping of particulate organic matter, near-bottom oxygen depletion and ammonium accumulation suggest that the vertical exchange of organic particles and dissolved oxygen is weakened at least temporarily. To better understand the causal links between the hydrodynamics and the oxygen and nutrient cycling in the deepened freshwater reach of the Elbe estuary, we establish a three-dimensional coupled hydrodynamical-biogeochemical model. The model demonstrates good skill in simulating the variability of the physical and biogeochemical parameters in the focal area. Coupled simulations reveal that this region is a hotspot of the degradation of diatoms and organic matter transported from the shallow productive upper estuary and the tidal weir. In summer, the water column weakly stratifies when at the bathymetric jump warmer water from the shallow upper estuary spreads over the colder water of the deepened mid reaches. Enhanced thermal stratification also occurs also in the narrow port basins and channels. Model results show intensification of the particle trapping due to the thermal gradients. The stratification also reduces the oxygenation of the near-bottom region and sedimentary layer inducing oxygen depletion and accumulation of ammonium. The study highlights that the vertical resolution is important for the understanding and simulation of estuarine ecological processes, because even weak stratification impacts the cycling of nutrients via modulation of the vertical mixing of oxygen, particularly in deepened navigation channels and port areas.

Antagonistic Interaction between Phosphinothricin and Nepeta rtanjensis Essential Oil Affected Ammonium Metabolism and Antioxidant Defense of Arabidopsis Grown In Vitro

Phosphinothricin (PPT) is one of the most widely used herbicides. PTT targets glutamine synthetase (GS) activity in plants, and its phytotoxicity is ascribed to ammonium accumulation and reactive oxygen species bursts, which drives rapid lipid peroxidation of cell membranes. In agricultural fields, PPT is extensively sprayed on plant foliage; however, a portion of the herbicide reaches the soil. According to the present study, PPT absorbed via roots can be phytotoxic to Arabidopsis, inducing more adverse effects in roots than in shoots. Alterations in plant physiology caused by 10 days exposure to herbicide via roots are reflected through growth suppression, reduced chlorophyll content, perturbations in the sugar and organic acid metabolism, modifications in the activities and abundances of GS, catalase, peroxidase, and superoxide dismutase. Antagonistic interaction of Nepeta rtanjensis essential oil (NrEO) and PPT, emphasizes the existence of complex control mechanisms at the transcriptional and posttranslational level, which result in the mitigation of PPT-induced ammonium toxicity and in providing more efficient antioxidant defense of plants. Simultaneous application of the two agents in the field cannot be recommended; however, NrEO might be considered as the PPT post-treatment for reducing harmful effects of herbicide residues in the soil on non-target plants.

Ammonium Accumulation Caused by Reduced Tonoplast V-ATPase Activity in Arabidopsis thaliana

Plant vacuoles are unique compartments that play a critical role in plant growth and development. The vacuolar H+-ATPase (V-ATPase), together with the vacuolar H+-pyrophosphatase (V-PPase), generates the proton motive force that regulates multiple cell functions and impacts all aspects of plant life. We investigated the effect of V-ATPase activity in the vacuole on plant growth and development. We used an Arabidopsisthaliana (L.) Heynh. double mutant, vha-a2 vha-a3, which lacks two tonoplast-localized isoforms of the membrane-integral V-ATPase subunit VHA-a. The mutant is viable but exhibits impaired growth and leaf chlorosis. Nitrate assimilation led to excessive ammonium accumulation in the shoot and lower nitrogen uptake, which exacerbated growth retardation of vha-a2 vha-a3. Ion homeostasis was disturbed in plants with missing VHA-a2 and VHA-a3 genes, which might be related to limited growth. The reduced growth and excessive ammonium accumulation of the double mutant was alleviated by potassium supplementation. Our results demonstrate that plants lacking the two tonoplast-localized subunits of V-ATPase can be viable, although with defective growth caused by multiple factors, which can be alleviated by adding potassium. This study provided a new insight into the relationship between V-ATPase, growth, and ammonium accumulation, and revealed the role of potassium in mitigating ammonium toxicity.

Modelling Silicate – Nitrate - Ammonium co-limitation of algal growth and the importance of bacterial remineralisation based on an experimental Arctic coastal spring bloom culture study

Arctic coastal ecosystems are rapidly changing due to climate warming, which makes modelling their productivity crucially important to better understand future changes. System primary production in these systems is highest during the pronounced spring bloom, typically dominated by diatoms. Eventually the spring blooms terminate due to silicon or nitrogen limitation. Bacteria can play an important role for extending bloom duration and total CO2 fixation through ammonium regeneration. Current ecosystem models often simplify the effects of nutrient co-limitations on algal physiology and cellular ratios and neglect bacterial driven regeneration, leading to an underestimation of primary production. Detailed biochemistry- and cell-based models can represent these dynamics but are difficult to tune in the environment. We performed a cultivation experiment that showed typical spring bloom dynamics, such as extended algal growth via bacteria ammonium remineralisation, and reduced algal growth and inhibited chlorophyll synthesis under silicate limitation, and gradually reduced nitrogen assimilation and chlorophyll synthesis under nitrogen limitation. We developed a simplified dynamic model to represent these processes. The model also highlights the importance of organic matter excretion, and post bloom ammonium accumulation. Overall, model complexity is comparable to other ecosystem models used in the Arctic while improving the representation of nutrient co-limitation related processes. Such model enhancements that now incorporate increased nutrient inputs and higher mineralization rates in a warmer climate will improve future predictions in this vulnerable system.

Excessive assimilation of ammonium by plastidic glutamine synthetase is a major cause of ammonium toxicity in Arabidopsis thaliana

Plants use nitrate and ammonium in the soil as their main nitrogen sources. Recently, ammonium has attracted attention due to evidence suggesting that, in C3 species, an elevated CO2 environment inhibits nitrate assimilation. However, high concentrations of ammonium as the sole nitrogen source for plants causes impaired growth, i.e. ammonium toxicity. Although ammonium toxicity has been studied for a long time, the primary cause remains to be elucidated. Here, we show that ammonium assimilation in plastids rather than ammonium accumulation is a primary cause for toxicity. Our genetic screen of ammonium-tolerant Arabidopsis lines with enhanced shoot growth identified plastidic GLUTAMINE SYNTHETASE 2 (GLN2) as the causal gene. Our reciprocal grafting of wild-type and GLN2 or GLN1;2-deficient lines suggested that shoot GLN2 activity results in ammonium toxicity, whilst root GLN1;2 activity prevents it. With exposure to toxic levels of ammonium, the shoot GLN2 reaction produced an abundance of protons within cells, thereby elevating shoot acidity and stimulating expression of acidic stress-responsive genes. Application of an alkaline ammonia solution to the toxic ammonium medium efficiently alleviated the ammonium toxicity with a concomitant reduction in shoot acidity. Consequently, we conclude that a primary cause of ammonium toxicity is acidic stress in the shoot. This fundamental insight provides a framework for enhanced understanding of ammonium toxicity in plants.