The energy cost of primary metabolism vacuole expansion: central to shape toamto leaf development under ammonium nutrition
- Autores: Theo Jean Poucet
- Directores de la Tesis: Daniel Marino (dir. tes.), Martine Dieuaide Noubhani (dir. tes.)
- Lectura: En la Universidad del País Vasco - Euskal Herriko Unibertsitatea ( España ) en 2020
- Idioma: inglés
- Tribunal Calificador de la Tesis: Dominique Rolin (presid.), Usue Perez Lopez (secret.), Nadine Paris (voc.), Celine Masclaux Daubresse (voc.), César Arrese-Igor Sánchez (voc.)
- Programa de doctorado: Programa de Doctorado en Agrobiología Ambiental por la Universidad del País Vasco/Euskal Herriko Unibertsitatea y la Universidad Pública de Navarra
- Materias:
- Enlaces
- Tesis en acceso abierto en: ADDI
- Resumen
Ammonium (NH4+) is a nitrogen source of great interest in the context of sustainable agriculture. Its application in the field together with nitrification inhibitors has been extensively proven efficient to limit detrimental N losses compared to the use of nitrate (N03). NH4+ is a common intermediate involved in numerous metabolic routes. However, high NH4 concentrations may lead to a stress situation provoking a set of symptoms collectively known as "ammonium syndrome" mainly characterized by growth retardation. Those symptoms are caused by a combination of, among others, a profound metabolic reprogramming, disruption of photosynthesis, pH deregulation and ion imbalance. Numerous studies have described the way plant copes to ammonium nutrition. However, the organ developmental stage has been generally neglected.To fill in this gap, in the first chapter we first aimed studying how the metabolism is adapted in function of the leaf position in the vertical axis of the tomato plants (Solanum lycopersicum) grown with NH4+, N03- or NH4N03 supply. To do so, we dissected leaf biomass composition and metabolism through a complete analysis of metabolites, ions and enzyme activities. The results showed that C and N metabolic adjustment in function of the nitrogen source was more intense in older leaves compared to younger ones. Importantly, we propose a trade-off between NH4+ accumulation and assimilation to preserve young leaves from ammonium stress. Besides, NH4+-fed plants exhibited a rearrangement of carbon skeletons with a higher energy cost respect to plants supplied with N03-. We explain such reallocation by the action of the biochemical pH-stat, to compensate the differential proton production that depends on the nitrogen form provided.Ammonium nutrition may limit cell expansion, suggesting that the cellular processes involved would be altered. Among others, cell growth is largely dependent of the internal pressure exerted on the cell wall by the vacuole. However, the role of the vacuole in ammonium stress has been rarely addressed. In the second chapter, we evaluated the effect of ammonium stress on leaf development with a special focus on vacuole expansion and metabolism. To carry out this aim, we monitored the leaf development from its appearance until its complete expansion in plants grown under NH4+ or NO/ as unique nitrogen source. Cytological analysis evidenced that the reduced cell expansion under ammonium nutrition was associated with smaller vacuole size. Besides, we reported an acidification of the vacuole of NH4+-fed plants compared to nitrate nutrition. Moreover, a model was built to predict the thermodynamic equilibrium of different soluble species across the tonoplast. The model was set up through an extensive reviewing of vacuolar transporters and integrated subcellular volumes, vacuolar electrochemical gradients and the formation of ionic complex in the vacuole to fit the subcellular concentration of ions, organic acids and sugars measured in the leaf. Further, predictions obtained with the model were cross validated with data from non-aqueous fractionation. Firstly, the entrance of solutes was higher in vacuoles of N03--fed leaves but was not associated with higher vacuolar osmolarity likely because of the adjustment of the vacuolar volume. In this sense, we proposed that the lack of malate in cells of ammonium-fed leaves was central in the limitation of vacuolar expansion. Secondly, we conclude that the energy cost of solute transport into the vacuole is higher under NH4+ based nutrition because of the higher electrochemical gradient generated by the proton pumps across tonoplast.This work highlights the importance of considering leaf phenological state when studying nitrogen metabolism. In addition, our integrated approach place cytosolic pH control and vacuole expansion in the center of tomato leaf adaptation to ammonium stress and pave the way for future studies in the field of ammonium nutrition.
