Plant ammonium sensitivity is associated with external pH adaptation, repertoire of nitrogen transporters, and nitrogen requirement.

Modern crops exhibit diverse sensitivities to ammonium as the primary nitrogen source, influenced by environmental factors such as external pH and nutrient availability. Despite its significance, there is currently no systematic classification of plant species based on their ammonium sensitivity. We conducted a meta-analysis of 50 plant species and present a new classification method based on the comparison of fresh biomass obtained under ammonium and nitrate nutrition. The classification uses the natural logarithm of the biomass ratio as the size effect indicator of ammonium sensitivity. This numerical parameter is associated with critical factors for nitrogen demand and form preference, such as Ellenberg indicators and the repertoire of nitrogen transporters for ammonium and nitrate uptake. Finally, a comparative analysis of the developmental and metabolic responses, including hormonal balance, is conducted in two species with divergent ammonium sensitivity values in the classification. Results indicate that nitrate has a key role in counteracting ammonium toxicity in species with a higher abundance of genes encoding NRT2-type proteins and fewer of those encoding the AMT2-type proteins. Additionally, the study demonstrates the reliability of the phytohormone balance and methylglyoxal content as indicators for anticipating ammonium toxicity.

Quantitative proteomics reveals the importance of nitrogen source to control glucosinolate metabolism in Arabidopsis thaliana and Brassica oleracea.

Accessing different nitrogen (N) sources involves a profound adaptation of plant metabolism. In this study, a quantitative proteomic approach was used to further understand how the model plant Arabidopsis thaliana adjusts to different N sources when grown exclusively under nitrate or ammonium nutrition. Proteome data evidenced that glucosinolate metabolism was differentially regulated by the N source and that both TGG1 and TGG2 myrosinases were more abundant under ammonium nutrition, which is generally considered to be a stressful situation. Moreover, Arabidopsis plants displayed glucosinolate accumulation and induced myrosinase activity under ammonium nutrition. Interestingly, these results were also confirmed in the economically important crop broccoli (Brassica oleracea var. italica). Moreover, these metabolic changes were correlated in Arabidopsis with the differential expression of genes from the aliphatic glucosinolate metabolic pathway. This study underlines the importance of nitrogen nutrition and the potential of using ammonium as the N source in order to stimulate glucosinolate metabolism, which may have important applications not only in terms of reducing pesticide use, but also for increasing plants' nutritional value.

Could ammonium nutrition increase plant C-sink strength under elevated CO2 conditions?

Atmospheric carbon dioxide (CO2) is increasing, and this affects plant photosynthesis and biomass production. Under elevated CO2 conditions (eCO2), plants need to cope with an unbalanced carbon-to-nitrogen ratio (C/N) due to a limited C sink strength and/or the reported constrains in leaf N. Here, we present a physiological and metabolic analysis of ammonium (NH4+)-tolerant pea plants (Pisum sativum L., cv. snap pea) grown hydroponically with moderate or high NH4+ concentrations (2.5 or 10 mM), and under two atmospheric CO2 concentrations (400 and 800 ppm). We found that the photosynthetic efficiency of the NH4+ tolerant pea plants remain intact under eCO2 thanks to the capacity of the plants to maintain the foliar N status (N content and total soluble proteins), and the higher C-skeleton requirements for NH4+ assimilation. The capacity of pea plants grown at 800 ppm to promote the C allocation into mobile pools of sugar (mainly sucrose and glucose) instead of starch contributed to balancing plant C/N. Our results also support previous observations: plants exposed to eCO2 and NH4+ nutrition can increase of stomatal conductance. Considering the C and N source-sink balance of our plants, we call for exploring a novel trait, combining NH4+ tolerant plants with a proper NH4+ nutrition management, as a way for a better exploitation of eCO2 in C3 crops.

The auxin-resistant dgt tomato mutant grows less than the wild type but is less sensitive to ammonium toxicity and nitrogen deficiency.

The low-auxin-sensitivity tomato mutant, dgt, despite displaying reduced plant growth, has been linked to greater resistance to N deficiency. This led us to test the role of auxin resistance of dgt in NH4+ toxicity and N deficiency, compared to wild type tomato (cv. Micro-Tom, MT), grown in hydroponic media. A completely randomized design with three replications in a 2 × 4 factorial scheme was adopted, corresponding to the two tomato genotypes (MT and dgt), involving four nutritional treatments: NO3- (5 mM); NH4+ (5 mM); NO3- (5 mM) plus exogenous auxin (10 µM IAA); and N omission. The results show that NH4+ was toxic to MT but not to dgt. Under N deficiency, MT displayed a lower shoot NO3- content, a lower photosynthetic rate, and a decrease in both shoot and root dry weight. However, in dgt, no difference was observed in shoot NO3- content and photosynthetic rate between plants grown on NO3- or under N deficiency. In addition, dgt showed an increase in shoot dry weight under N deficiency. We highlight the role of auxin resistance in the adaptation of plants to NH4+ toxicity and N deficiency.

Multi-omic and physiologic approach to understand Lotus japonicus response upon exposure to 3,4 dimethylpyrazole phosphate nitrification inhibitor.

Nitrogen fertilization is a major force in global greenhouse gases emissions and causes environmental contamination through nitrate leaching. The use of nitrification inhibitors has been proven successful to mitigate these effects. However, there is an increasing concern about the undesired effects that their potential persistence in the soil or accumulation in plants may provoke. In this study, we first exposed Lotus japonicus plants to high amounts of 3,4 dimethylpyrazole phosphate (DMPP) and 2-(N-3,4-dimethyl-1H-pyrazol-1-yl) succinic acid isomeric mixture (DMPSA) nitrification inhibitors. Exposure to doses higher than 1 mg·L-1 provoked DMPP accumulation mostly in the aerial part, while DMPSA was only detected from 10 mg·L-1 and nearly no translocation. To evaluate the effect that DMPP accumulation in leaves may provoke on plant performance we combined a transcriptome, proteome, and physiological analysis in plants treated with 10 mg/ L of DMPP. This treatment provoked changes in the expression of 229 genes and 59 proteins. Overall, we evidence that when DMPP accumulates in leaves it induces stress responses, notably provoking changes in cell redox balance, hormone signaling, protein synthesis and turnover and carbon and nitrogen metabolism.

3,4-Dimethylpyrazole phosphate and 2-(N-3,4-dimethyl-1H-pyrazol-1-yl) succinic acid isomeric mixture nitrification inhibitors: Quantification in plant tissues and toxicity assays.

Nitrification inhibitors are used to maintain ammonium available in the soil for longer periods while reducing nitrate leaching and N2O emission. In this work we evaluated the potential toxicity effects of 3,4-dimethylpyrazole phosphate (DMPP) and 2-(N-3,4-dimethyl-1H-pyrazol-1-yl) succinic acid isomeric mixture (DMPSA) nitrification inhibitors. In order to determine the potential plant capacity to take up and translocate these inhibitors, we grew clover plants in hydroponic conditions and we developed a novel methodology for extracting DMPP and DMPSA that we quantified by HPLC. In addition, we also did toxicity bioassays: seed germination and Vibrio fischeri test. When clover was exposed to high amounts of nitrification inhibitors, plants accumulated DMPP, predominantly in leaves, and also DMPSA that preferentially accumulated in roots. These inhibitors did not provoke phytotoxicity at the equivalent of the maximum amount estimated in agriculture (0.5mg/kg soil). DMPP only provoked detrimental effects in plants at very high dose (100mg/kg soil). Interestingly, DMPSA was innocuous.

Overexpression of a pine Dof transcription factor in hybrid poplars: A comparative study in trees growing under controlled and natural conditions.

In this work, the role of the pine transcriptional regulator Dof 5 in carbon and nitrogen metabolism has been examined in poplar trees. The overexpression of the gene and potential effects on growth and biomass production were compared between trees growing in a growth chamber under controlled conditions and trees growing in a field trial during two growth seasons. Ten-week-old transgenic poplars exhibited higher growth than untransformed controls and exhibited enhanced capacity for inorganic nitrogen uptake in the form of nitrate. Furthermore, the transgenic trees accumulated significantly more carbohydrates such as glucose, fructose, sucrose and starch. Lignin content increased in the basal part of the stem likely due to the thicker stem of the transformed plants. The enhanced levels of lignin were correlated with higher expression of the PAL1 and GS1.3 genes, which encode key enzymes involved in the phenylalanine deamination required for lignin biosynthesis. However, the results in the field trial experiment diverged from those observed in the chamber system. The lines overexpressing PpDof5 showed attenuated growth during the two growing seasons and no modification of carbon or nitrogen metabolism. These results were not associated with a decrease in the expression of the transgene, but they can be ascribed to the nitrogen available in the field soil compared to that available for growth under controlled conditions. This work highlights the paramount importance of testing transgenic lines in field trials.

The physiological implications of urease inhibitors on N metabolism during germination of Pisum sativum and Spinacea oleracea seeds.

The development of new nitrogen fertilizers is necessary to optimize crop production whilst improving the environmental aspects arising from the use of nitrogenous fertilization as a cultural practice. The use of urease inhibitors aims to improve the efficiency of urea as a nitrogen fertilizer by preventing its loss from the soil as ammonia. However, although the action of urease inhibitors is aimed at the urease activity in soil, their availability for the plant may affect its urease activity. The aim of this work was therefore to evaluate the effect of two urease inhibitors, namely acetohydroxamic acid (AHA) and N-(n-butyl) thiophosphoric triamide (NBPT), on the germination of pea and spinach seeds. The results obtained show that urease inhibitors do not affect the germination process to any significant degree, with the only process affected being imbibition in spinach, thus also suggesting different urease activities for both plants. Our findings therefore suggest an activity other than the previously reported urolytic activity for urease in spinach. Furthermore, of the two inhibitors tested, NBPT was found to be the most effective at inhibiting urease activity, especially in pea seedlings.

Short term physiological implications of NBPT application on the N metabolism of Pisum sativum and Spinacea oleracea.

The application of urease inhibitors in conjunction with urea fertilizers as a means of reducing N loss due to ammonia volatilization requires an in-depth study of the physiological effects of these inhibitors on plants. The aim of this study was to determine how the urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT) affects N metabolism in pea and spinach. Plants were cultivated in pure hydroponic culture with urea as the sole N source. After 2 weeks of growth for pea, and 3 weeks for spinach, half of the plants received NBPT in their nutrient solution. Urease activity, urea and ammonium content, free amino acid composition and soluble protein were determined in leaves and roots at days 0, 1, 2, 4, 7 and 9, and the NBPT content in these tissues was determined 48h after inhibitor application. The results suggest that the effects of NBPT on spinach and pea urease activity differ, with pea being most affected by this treatment, and that the NBPT absorbed by the plant caused a clear inhibition of the urease activity in pea leaf and roots. The high urea concentration observed in leaves was associated with the development of necrotic leaf margins, and was further evidence of NBPT inhibition in these plants. A decrease in the ammonium content in roots, where N assimilation mainly takes place, was also observed. Consequently, total amino acid contents were drastically reduced upon NBPT treatment, indicating a strong alteration of the N metabolism. Furthermore, the amino acid profile showed that amidic amino acids were major components of the reduced pool of amino acids. In contrast, NBPT was absorbed to a much lesser degree by spinach plants than pea plants (35% less) and did not produce a clear inhibition of urease activity in this species.