Browsing by Subject "Ammonium"

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Developing indicators and characterizing direct and residual effects of biological nitrification inhibition (BNI) by the tropical forage grass Brachiaria humidicola
(2018) Karwat, Hannes; Cadisch, Georg
Nitrogen (N) losses from agroecosystems harm the environment via increased nitrate (NO3-) amounts in water-bodies and nitrous oxide (N2O) emissions to the atmosphere. Bacteria and archaea oxidize ammonium (NH4+) to NO3- under aerobic conditions. Furthermore, under mainly anaerobic conditions, microbial denitrification reduces NO3- to gaseous N forms. The tropical forage grass Brachiaria humidicola (Rendle) Schweick (Bh) has been shown to reduce soil microbial nitrification via root derived substances. Therefore, biological nitrification inhibition (BNI) by Bh might contribute to reduction of N losses from agroecosystems. The present doctoral thesis aimed at assessing the potential of the actual BNI by Bh, as well as the residual BNI effect with new developed methodologies. The overall research was based on the following major objectives: (1) characterization of the residual BNI effect by Bh on recovery of N by subsequent cropped maize (Zea mays L.) under different N fertilization rates; (2) investigate if low enzymatic nitrate reductase activity (NRA) in leaves of Bh is linked to reduced NO3- nutrition by effective BNI; (3) identify a possible link between plant delta 15N of Bh and the BNI effect of different Bh genotypes on nitrification, plant N uptake and NO3- leaching losses. The overall objective was to use and test new methodologies with a minimum of disturbance of the plant-soil system, to characterize BNI of different Bh genotypes in greenhouse and field studies. The first research study focused on the investigation of a potential residual BNI effect of a converted long-term Bh pasture on subsequent maize cropping, where a long-term maize monocrop field served as control. The residual BNI effect was characterized in terms of enhanced maize grain yield, total N uptake and 15N (labeled) fertilizer recovery. Furthermore, the impact of residual BNI effect on soil N dynamics was investigated. The residual BNI effect was confirmed for the first maize crop season after pasture conversion on the basis of lower nitrification in incubation soil, higher total N uptake and higher maize grain yields. However, the residual BNI effect did not result in higher 15N fertilizer uptake or reduced 15N fertilizer losses, nor in reduced N20 emissions. Applied N was strongly immobilized due to long-term root turnover effects, while a significant residual BNI effect from Bh prevented re-mineralized N from rapid nitrification resulting in improved maize performance. A significant residual Bh BNI effect was evident for less than one year only. In the second research study it was the aim to verify the potential of nitrate reductase activity (NRA) as a proxy for the detection of in vivo performance of BNI by selected Bh accessions and genotypes grown under contrasting fertilization regimes. NRA was detected in Bh leaves rather than in roots, regardless of NO3- availability. Leaf NRA correlated with NO3- contents in soils and stem sap of contrasting Bh genotypes substantiating its use as a proxy of in vivo performance of BNI. The leaf NRA assay facilitated a rapid screening of contrasting Bh genotypes for their differences in in vivo performance of BNI under field and greenhouse conditions; but inconsistency of the BNI potential by selected Bh genotypes was observed. The third research study emphasized to link the natural abundance of delta 15N in Bh plants with reduced NO3- losses and enhanced N uptake due to BNI. Increased leached NO3- was positively correlated to rising delta 15N in Bh grass, whereas the correlation between plant N uptake and plant delta 15N was inverse. Long-term field cultivation of Bh decreased nitrification in incubated soil, whereas delta 15N of Bh declined and plant N% rose over time. Delta 15N of Bh correlated positively with assessed nitrification rates in incubated soil. It was concluded that decreasing delta 15N of Bh over time reflects the long-term effect of BNI linked to lower NO3- formation and reduced NO3- leaching, and that generally higher BNI activity of Bh is indicated by lower delta 15N plant values. Within the framework of this thesis, a residual BNI effect by Bh on maize cropping could be confirmed for one season due to the combined methodological approaches of soil incubation and 15N recovery. The development of the NRA assay for sampled Bh leaves was validated as a rapid and reliable method linked to the actual soil nitrification after NH4+ fertilizer supply. Consequently, the assay could be used for both greenhouse and field studies as BNI proxy. The gathered data from the third study indicated that decreasing delta 15N of Bh over time reflects the long-term effect of BNI linked to lower NO3- formation and reduced NO3- leaching, and that generally higher BNI activity of Bh is indicated by lower delta 15N plant values. Consequently, it was suggested that delta 15N of Bh could serve as an indicator of cumulative NO3- losses. Overall, this doctoral thesis suggests the depressing effect on nitrification by Bh might be a combined effect by BNI and fostered N immobilization. Furthermore, BNI by Bh might be altered by different factors such as soil type, plant age and root morphology of the genotypes. Finally, future studies should consider that Bh genotypes express their respective BNI potential differently under contrasting conditions.
Iron and ammonium sensing differentially modulate root plasticity in Arabidopsis thaliana
(2010) Lima, Joni Esrom; von Wirén, Nicolaus
Modulation of root system architecture by plants has an impact on water, nutrient acquisition and anchorage during plant development. In a given environment, root plasticity is a favorable feature to react according to abiotic and biotic factors. Under nutrient limited conditions, the root plasticity is essential for a better soil volume exploitation. This response can vary according to the plant species and the given environment in which they evolved. Moreover, nutrient mobility in the soil plays an important role for the response of plants to nutrient limitation. Thus, root plasticity is a nutrient-specific response during plant development. In fact research on the effect of nutrient availability on root system architecture is scarce. Furthermore, the mechanism how plants sense nutrients and the signaling upon nutrient availability remains a challenge. Therefore, identification of which nutrient can affect the root system architecture and investigating the molecular components involved in the signaling pathway is certainly relevant for agronomical practices. The first part of the present work aimed to identify how the root architecture is affected by iron (Fe) supply. Due to the low mobility of Fe in soils, the morphological response of lateral roots from Arabidopsis plants to localized Fe supply and its regulation were investigated. Increasing Fe concentrations in a homogenous or localized supply on separated agar plates enhanced lateral root number in a similar manner. Lateral root length, however, was twofold higher under localized relative to homogenous Fe supply. With further increasing Fe concentrations lateral root length was repressed even though shoot growth was unaffected. In the Fe uptake-defective mutant irt1, the formation of lateral roots required higher local Fe supplies, which restored wild type levels only with respect to the number but not to the length of lateral roots. Moreover, IRT1 transcript levels were strongly enhanced under localized Fe supply. In the frd3-1 mutant, which is defective in root-to-shoot translocation of Fe, lateral root development was similar to wild type plants although frd3-1 shoots were Fe deficient. These results show a differential regulation of lateral root initiation and elongation in response to localized Fe supply and that lateral root elongation is under control of a local rather than a systemic regulatory loop involving the high-affinity Fe transporter IRT1. In the second part of the thesis, a remarkable and an unknown feature of root morphology dependent on localized ammonium supply is described. Arabidopsis plants were able to increase lateral root initiation and higher-order lateral root branching. Since ammonium-stimulated lateral root number or density decreased after ammonium or glutamine supply to a separate root fraction and did not correlate with cumulative uptake of 15N-labeled ammonium, lateral root branching was not purely due to a nutritional effect but most likely a sensing event. Moreover, a detailed investigation has shown that ammonium and nitrate co-ordinate root morphology in an additive and complementary way. By a genetic approach, the ammonium-induced lateral root branching was demonstrated to be dependent on AMT1;3 activity in the root.
Leaf gas exchange of lowland rice in response to nitrogen source and vapor pressure deficit
(2021) Vu, Duy Hoang; Stürz, Sabine; Pieters, Alejandro; Asch, Folkard
Background: In anaerobic lowland fields, ammonium (NH4+) is the dominant form of nitrogen (N) taken up by rice plants, however, with the large expansion of water-saving irrigation practices, nitrification is favored during drained periods, leading to an increased availability of nitrate (NO3−). Aim: Since the uptake and assimilation of the two N-sources differ in their demand of pho- tosynthates, leaf gas exchange may be subject to adjustments in response to N-sources, particularly at high evaporative demand, when stomatal conductance (gs ) is very sensitive. Methods: Three experiments were carried out to study leaf gas exchange of various low- land rice varieties in response to N-source at low and high vapor pressure deficit (VPD). In the first experiment, seedlings of 12 rice varieties were grown at high VPD for 3 weeks. From this, four rice varieties differing in gs and CO2 assimilation rate (A) were selected and grown for 2 weeks at low VPD, and after that, they were shifted to high VPD for 1 week, whereas in the third experiment, the same varieties were grown separately at low and high VPD conditions for 2 weeks. In all three experiments, plants were grown hydroponi- cally in nutrient solution with N-sources as sole NH4+ or NO3−. Results: At high VPD, NO3− nutrition led to a higher gs and A in four out of 12 vari- eties (IR64, BT7, NU838, and Nipponbare) relative to NH4+ nutrition, while no effect was observed at low VPD or after a short-term exposure to high VPD. Further, varieties with a high intrinsic water-use efficiency (WUEi; IR64 and BT7) showed the strongest response to N-source. Higher gs was partially supported by increased root/shoot ratio, but could not be fully explained by the measured parameters. However, higher A in NO3−-fed plants did not always result in increased plant dry matter, which is probably related to the higher energy demand for NO3− assimilation. Our results suggest that at high VPD, NO3− nutri- tion can improve leaf gas exchange in varieties having a high WUEi, provided a sufficient water supply. Conclusion: Therefore, intensified nitrification under water-saving irrigation measures may improve leaf gas exchange and the growth of rice plants under high transpirational demand. However, choice of variety seems crucial since large varietal differences were observed in response to N-source. Further, breeding strategies for genotypes adapted to aerobic soil conditions should consider responses to NO3−, potentially using gas exchange measurements as a screening tool.
Plant ammonium transporter (AMT) integration in regulatory networks
(2016) Straub, Tatsiana; Ludewig, Uwe
Ammonium is a ubiquitous key nutrient in agricultural soils and the preferred nitrogen source for plants. However, excessive ammonium accumulation represses plant growth and development. Ammonium is taken up by plant cells via high-affinity ammonium transporters (AMTs). Six AMT genes were identified in Arabidopsis, which are separated in two distinct clades, five AMT1s and one AMT2. In the plasma membrane, AMT proteins form homo- and heterotrimers with extra-cytoplasmic N-termini and cytoplasmic C-termini. In addition to transcriptional and post-transcriptional control of AMTs by ammonium, phosphorylation in the C-terminus serves as a rapid allosteric switch of the AMT activity and prevents further internal ammonium accumulation. In a physiological screen, a kinase (CIPK23) was identified, which directly regulates ammonium transport activity under high-NH4+ conditions. Interestingly, CIPK23 is already known to regulate nitrate and potassium uptake in roots. Lesion of the CIPK23 gene significantly increased ammonium uptake, but caused growth inhibition. As expected, cipk23 plants were also limited in potassium accumulation, but high potassium availability failed to rescue the cipk23 phenotype. Furthermore, cipk23 plants were more susceptible to methylammonium (MeA), a non-metabolizable analogue of ammonium. The sensitivity to MeA was lost upon genetic suppression of AMT1 genes in the cipk23 background. The data suggest that CIPK23 directly phosphorylates AMT1s in a complex with CBL1 (calcineurin B-like protein) and thereby regulates transport activity. The expression of the CIPK23 and the CBL1 genes were ammonium-dependent and increased when N-starved plants were resupplied with ammonium. Furthermore, cbl1 mutants had enhanced NH4+ accumulation; this phenocopies the larger ammonium uptake in the cipk23 loss-of-function mutant. In vivo experiments demonstrated bimolecular interaction between CIPK23, AMT1;1, and AMT1;2, but not with AMT2;1, suggesting direct phosphorylation of AMT1-type ammonium transporters by CIPK23. However, Western blot analysis with the cipk23 mutant suggested that the loss of the kinase was not sufficient to completely abolish AMT1;1 and AMT1;2 phosphorylation, indicating several independent pathways to regulate ammonium transport activity in AMT trimers. The data identify complex post-translational regulation of ammonium transporters via the CBL1–CIPK23 pathway, which ensures reduction of AMT1 activity and suppression of ammonium uptake under high external NH4+ concentrations.
Regulation of ammonium transport in Arabidopsis thaliana
(2022) Ganz, Pascal; Ludewig, Uwe
The overarching question of this thesis deals with how plants ensure the selective uptake of ammonium while maintaining ion and pH homeostasis. A key component of this is ammonium transporters (AMTs) with high affinity towards their substrate, which are at the same time part of a multilayered protection system against uncontrolled ammonium influx. Conserved protein sequences inside the transporter were analyzed as well as the regulatory system based on post-translational modification of the transporter.