Combined Allosteric Responses Explain the Bifurcation in Non-Linear Dynamics of N Root Fluxes Under Nutritional Steady-State Conditions for Nitrate.

With regard to thermodynamics out of equilibrium, seedlings are open systems that dissipate energy towards their environment. Accordingly, under nutritional steady-state conditions, changes in external concentrations of one single ion provokes instability and reorganization in the metabolic and structure/architecture of the seedling that is more favorable to the fluxes of energy and matter. This reorganization is called a bifurcation and is described in mathematics as a non-linear dynamic system.

Inhibition of Aminotransferases by Aminoethoxyvinylglycine Triggers a Nitrogen Limitation Condition and Deregulation of Histidine Homeostasis That Impact Root and Shoot Development and Nitrate Uptake.

Although AVG (aminoethoxyvinylglycine) is intensely used to decipher signaling in ethylene/indol-3-acetic acid (IAA) interactions on root morphogenesis, AVG is not a specific inhibitor of aminocyclopropane-1-carboxylate synthase () and tryptophan aminotransferase () and tryptophan aminotransferase related () activities since it is able to inhibit several aminotransferases involved in N metabolism. Indeed, 1 mM glutamate (Glu) supply to the roots in plants treated with 10 μM AVG partially restores the root growth. Here, we highlight the changes induced by AVG and AVG + Glu treatments on the N metabolism impairment and root morphogenetic program.

Changes in NO Availability and Transpiration Rate Are Associated With a Rapid Diurnal Adjustment of Anion Contents as Well as N and Water Fluxes Between the Roots and Shoots.

Understanding interactions between water and nitrate fluxes in response to nitrate availability and transpiration rate is crucial to select more efficient plants for the use of water and nitrate. Some of these interactions were investigated in intact plants by combining a non-destructive gravimetric device with NO labeling. The set-up allowed high-resolution measurement of the effects of a cross-combination of two concentrations of KNO or KCl (0.

The Thermodynamic Flow-Force Interpretation of Root Nutrient Uptake Kinetics: A Powerful Formalism for Agronomic and Phytoplanktonic Models.

The ion influx isotherms obtained by measuring unidirectional influx across root membranes with radioactive or stable tracers are mostly interpreted by enzyme-substrate-like modeling. However, recent analyses from ion transporter mutants clearly demonstrate the inadequacy of the conventional interpretation of ion isotherms. Many genetically distinct carriers are involved in the root catalytic function.

Fine-tuning of root elongation by ethylene: a tool to study dynamic structure-function relationships between root architecture and nitrate absorption.

Background Recently developed genetic and pharmacological approaches have been used to explore NO3-/ethylene signalling interactions and how the modifications in root architecture by pharmacological modulation of ethylene biosynthesis affect nitrate uptake. Key Results Structure-function studies combined with recent approaches to chemical genomics highlight the non-specificity of commonly used inhibitors of ethylene biosynthesis such as AVG (l-aminoethoxyvinylglycine). Indeed, AVG inhibits aminotransferases such as ACC synthase (ACS) and tryptophan aminotransferase (TAA) involved in ethylene and auxin biosynthesis but also some aminotransferases implied in nitrogen (N) metabolism.

Breaking conceptual locks in modelling root absorption of nutrients: reopening the thermodynamic viewpoint of ion transport across the root.

Background: The top-down analysis of nitrate influx isotherms through the Enzyme-Substrate interpretation has not withstood recent molecular and histochemical analyses of nitrate transporters. Indeed, at least four families of nitrate transporters operating at both high and/or low external nitrate concentrations, and which are located in series and/or parallel in the different cellular layers of the mature root, are involved in nitrate uptake. Accordingly, the top-down analysis of the root catalytic structure for ion transport from the Enzyme-Substrate interpretation of nitrate influx isotherms is inadequate.

An updated model for nitrate uptake modelling in plants. II. Assessment of active root involvement in nitrate uptake based on integrated root system age: measured versus modelled outputs.

Background and Aims An updated version of a mechanistic structural-functional model was developed to predict nitrogen (N) uptake throughout the growth cycle by a crop of winter oilseed rape, Brassica napus, grown under field conditions. Methods The functional component of the model derives from a revisited conceptual framework that combines the thermodynamic Flow-Force interpretation of nitrate uptake isotherms and environmental and in planta effects on nitrate influx. Estimation of the root biomass (structural component) is based upon a combination of root mapping along the soil depth profile in the field and a relationship between the specific root length and external nitrate concentration.

An updated model for nitrate uptake modelling in plants. I. Functional component: cross-combination of flow-force interpretation of nitrate uptake isotherms, and environmental and in planta regulation of nitrate influx.

Background And Aims: In spite of major breakthroughs in the last three decades in the identification of root nitrate uptake transporters in plants and the associated regulation of nitrate transport activities, a simplified and operational modelling approach for nitrate uptake is still lacking. This is due mainly to the difficulty in linking the various regulations of nitrate transport that act at different levels of time and on different spatial scales.

Methods: A cross-combination of a Flow-Force approach applied to nitrate influx isotherms and experimentally determined environmental and in planta regulation is used to model nitrate in oilseed rape, Brassica napus.

Modulation of ethylene biosynthesis by ACC and AIB reveals a structural and functional relationship between the K15NO3 uptake rate and root absorbing surfaces.

The modification of root traits in relation to nitrate uptake represents a source for improvement of nitrogen uptake efficiency. Because ethylene signalling modulates growth of exploratory and root hair systems more rapidly (minutes to hours) than nitrate signalling (days to weeks), a pharmacological approach was used to decipher the relationships between root elongation and N uptake. Rape seedlings were grown on agar plates supplied with 1mM K(15)NO3 and treated with different concentrations of either the ethylene precursor, ACC (0.

In low transpiring conditions, uncoupling the BnNrt2.1 and BnNrt1.1 NO 3(-) transporters by glutamate treatment reveals the essential role of BnNRT2.1 for nitrate uptake and the nitrate-signaling cascade during growth.

In plants, the nitrate transporters, NRT1.1 and NRT2.1, are mainly responsible for nitrate uptake.

In low transpiring conditions, nitrate and water fluxes for growth of B. napus plantlets correlate with changes in BnNrt2.1 and BnNrt1.1 transporter expression.

We analyzed how changes in BnNrt nitrate transporter gene expression induced by nitrate are associated with morphological changes in plantlets and osmotic water flow for growth. We hypothesized that in a Petri dish system, reduction in transpiration should induce conditions where nitrate and water fluxes for growth depend directly on nitrate transporter activity and nitrate signaling. Rape seedlings growing on agar plates were supplied with increasing external K (15)NO 3 concentrations from 0.

Ethylene modifies architecture of root system in response to stomatal opening and water allocation changes between root and shoot.

Ethylene plays a key role in the elongation of exploratory and root hair systems in plants, as demonstrated by pharmacological modulation of the activity of ethylene biosynthesis enzymes: ACC synthase (ACS) and ACC oxidase (ACO). Thus, treatments with high concentrations (10 microM) of aminoethoxyvinylglycine (AVG, inhibitor of ACS) and 1-aminocyclopropane carboxylic acid (ACC, ethylene precursor, ACO activator) severely decrease the elongation of the exploratory root system but induce opposite effects on the root hair system: root hair length and numbers were increased in seedlings treated with ACC, whereas they were reduced in seedlings treated with AVG. Until now, such elongation changes of root architecture had not been questioned in terms of nitrate uptake.

Elongation changes of exploratory and root hair systems induced by aminocyclopropane carboxylic acid and aminoethoxyvinylglycine affect nitrate uptake and BnNrt2.1 and BnNrt1.1 transporter gene expression in oilseed rape.

Ethylene is a plant hormone that plays a major role in the elongation of both exploratory and root hair systems. Here, we demonstrate in Brassica napus seedlings that treatments with the ethylene precursor, aminocyclopropane carboxylic acid (ACC) and the ethylene biosynthesis inhibitor, aminoethoxyvinylglycine (AVG), cause modification of the dynamic processes of primary root and root hair elongation in a dose-dependent way. Moreover, restoration of root elongation in AVG-treated seedlings by 1 mm l-glutamate suggested that high concentrations of AVG affect root elongation through nonoverlapping ethylene metabolic pathway involving pyridoxal 5'-P-dependent enzymes of nitrate (N) metabolism.

Characterization of proteins secreted during maize microspore culture: arabinogalactan proteins (AGPs) stimulate embryo development.

To study molecules secreted from cultured plant cells that promote development, maize microspores were transferred into culture and the conditioned media were collected over time and analysed. Electrophoresis indicated that both non-glycosylated and glycosylated proteins including arabinogalactan proteins (AGPs) appeared in the medium and their concentration increased during the time of culture. The development of embryos was correlated with the presence of specific extracellular proteins, using an experimental system based on a tunicamycin inhibition test.

Oxidative burst and expression of germin/oxo genes during wounding of ryegrass leaf blades: comparison with senescence of leaf sheaths.

Two bursts of H(2)O(2) production have been detected by in situ 3,3'-diaminobenzidine (DAB) staining after cutting of Lolium perenne L. leaf blades. The first burst, which occurred immediately after wounding was inhibited by Na-diethydithiocarbamate (DIECA), a Cu/Zn-superoxide dismutase (SOD) inhibitor.

Modeling nitrogen uptake in oilseed rape cv Capitol during a growth cycle using influx kinetics of root nitrate transport systems and field experimental data.

The use of kinetic equations of NO3- transport systems in oilseed rape (Brassica napus), determined by 15NO3- labeling under controlled conditions, combined with experimental field data from the INRA-Châlons rape database were used to model NO3- uptake during the plant growth cycle. The quantitative effects of different factors such as day/night cycle, ontogenetic stages, root temperature, photosynthetically active radiation, and soil nitrate availability on different components of the constitutive high-affinity transport systems, constitutive low-affinity transport systems, inducible low-affinity transport systems, and inducible high-affinity transport systems of nitrate were then determined to improve the model's predictions. Simulated uptake correlated well with measured values of nitrogen (N) uptake under field conditions for all N fertilization rates tested.

Effects of nitrate pulses on BnNRT1 and BnNRT2 genes: mRNA levels and nitrate influx rates in relation to the duration of N deprivation in Brassica napus L.

A de-repression mechanism based on the disappearance of 'signals' down-regulating N transporter activity has been proposed in the literature to explain the transient increase of NO(3)(-) uptake by the roots following N deprivation in higher plants. This hypothesis was investigated at the physiological and molecular levels by measuring NO(3)(-) influx into roots of Brassica napus L. grown under low or high external concentrations of KNO(3) following N deprivation.