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Total Pages: 172 Number of Pages: 151 Missing Page: 146 Extra Pages: 131a and Data Source / The study presents models of portfolio behaviour for four industries, viz., cotton textiles, sugar, transport equipment and nonelectrical machinery. The first two industries are consumer goods industries and produce to stock. The other two are capital goods industries and produce to order. The study uses econometric techniques to estimate influence of variables explaining changes in capital stock, inventories, accounts receivable, cash balances and government bonds and also the composition of liabilities (debt and equity). / Thesis / Doctor of Philosophy (PhD)

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Pas de résumé en français / In plants, long distance transport of sugars from photosynthetic source leaves to sink organs comprises different crucial steps depending on the species and organ types. Sucrose, the main carbohydrate for long distance transport is synthesized in the mesophyll and then loaded into the phloem. After long distance transport through the phloem vessels, sucrose is finally unloaded towards sink organs. Alternatively, sugar can also be transferred to non‐plant sinks and plant colonization by heterotrophic organisms increases the sink strength and creates an additional sugar demand for the host plant. These sugar fluxes are coordinated by transport systems. Main sugar transporters in plants comprise sucrose (SUTs) and monosaccharide (MSTs) transporters which constitute key components for carbon partitioning at the whole plant level and in interactions with fungi. Although complete SUTs and MSTs gene families have been identified from the reference Dicot Arabidopsis thaliana and Monocot rice (Oriza sativa), sugar transporter families of the leguminous plant Medicago truncatula, which represents a widely used model for studying plant-fungal interactions in arbuscular mycorrhiza (AM), have not yet been investigated.With the recent completion of the M. truncatula genome sequencing as well as the release of transcriptomic databases, monosaccharide and sucrose transporter families of M. truncatula were identified and now comprise 62 MtMSTs and 6 MtSUTs. I focused on the study of the newly identified MtSUTs at a full family scale; phylogenetic analyses showed that the 6 members of the MtSUT family distributed in all three Dicotyledonous SUT clades; they were named upon phylogenetic grouping into particular clades: MtSUT1-1, MtSUT1-2, MtSUT1-3, MtSUT2, MtSUT4-1 and MtSUT4-2. Functional analyses by yeast complementation and expression profiles obtained by quantitative RT-PCR revealed that MtSUT1-1 and MtSUT4-1 are H+/sucrose symporters and represent key members of the MtSUT family. Conservation of transport capacity between orthologous leguminous proteins, expression profiles and subcellular localization compared to previously characterized plant SUTs indicate that MtSUT1-1 is the main protein involved in phloem loading in source leaves whilst MtSUT4-1 mediates vacuolar sucrose export for remobilization of intracellular reserve.The AM symbiosis between plants and fungi from the phylum Glomeromycota is characterized by trophic exchanges between the two partners. The fungus supplies the autotrophic host with nutrients and thereby promotes plant growth. In return, the host plant provides photosynthate (sugars) to the heterotrophic symbiont. Here, sugar fluxes from plant source leaves towards colonized sink roots in the association between the model leguminous plant M. truncatula and the arbuscular mycorrhizal fungus (AMF) Glomus intraradices were investigated. Sugar transporter candidates from both the plant and fungal partners presenting differential expression profiles using available transcriptomic tools were pinpointed. Gene expression profiles of MtSUTs and sugar quantification analyses upon high and low phosphorus nutrient supply and inoculation by the AMF suggest a mycorrhiza-driven stronger sink in AM roots with a fine-tuning regulation of MtSUT gene expression. Conserved regulation patterns were observed for orthologous SUTs in response to colonization by glomeromycotan fungi.In parallel, a non-targeted strategy consisting in the development of a M. truncatula - G. intraradices expression library suitable for yeast functional complementation and screening of symbiotic marker genes, similar to the approach that led to the identification of the first glomeromycotan hexose transporter (GpMST1), has been developed in this study. [...]

Dans son environnement la plante est confrontée à une variété de microorganismes bénéfiques, neutres et pathogènes, qui sont fortement dépendants des ressources carbonées qu’elle libère dans le sol. Le transport de sucres, processus clé de la physiologie de la plante, est essentiel pour les interactions plantes-microorganismes et leur devenir. Au cours de l'évolution, les plantes ont acquis des mécanismes leur permettant de percevoir les signaux microbiens du milieu extérieur, et aboutissant à la transduction d’un signal spécifique puis à des réponses biologiques adaptées (défense versus mutualisme) à la stratégie du microorganisme. Ces réponses assurent la survie et le développement des plantes. Mes travaux de thèse, menés avec un système « d’interaction » simplifié, contribuent à une meilleure compréhension des mécanismes sous-jacents au déterminisme des interactions plantes-microorganismes. Ce système a permis d’étudier, sur des suspensions cellulaires de N. tabacum, les réponses cellulaires précoces déclenchées suite à la perception de molécules microbiennes provenant de microorganismes à stratégie pathogène avirulent ou à stratégie mutualiste. Nous avons mesuré des évènements de signalisation et des flux de sucres induits en réponse à ces molécules microbiennes. Nos résultats ont mis en évidence que les chitotétrasaccharides (CO4), sécrétés par les champignons mycorhiziens à arbuscules dans les stades pré-symbiotiques de l’interaction, mobilisent les mêmes événements de signalisation précoce (H2O2 dépendant de la protéine rbohD, Ca2+ cytosolique, activation de MAPK) que la cryptogéine, un éliciteur des réactions de défense ; mais avec des réponses différentes en terme d’intensité et de cinétique. Les CO4 et la cryptogéine ont par ailleurs montré des impacts distincts sur les flux de sucres et l’expression de transporteurs impliqués. En complément nous avons montré un effet de la modification de la dynamique membranaire associée à la clathrine sur des évènements de signalisation déclenchés par la cryptogéine, ainsi que dans les flux entrants de sucres et l’expression de gènes de transporteurs de sucres. Enfin, l’analyse in silico de l’interactome de transporteurs de sucres chez la plante modèle A. thaliana, nous a permis d’apporter des connaissances supplémentaires quant aux évènements de régulations des transporteurs de sucres et l’identification de protéines régulatrices putatives en interaction avec ces derniers. L’ensemble de ces travaux ouvrent la voie à de nouvelles recherches visant à élucider les mécanismes cellulaires et moléculaires impliqués dans la mise en place des interactions entre plantes et microorganismes. / In their natural environment plants are in close interaction with beneficial, neutral, or pathogenic microbes, which are highly dependent on carbon resources exuded by plant roots. Sugar transport, which is a key process of plant physiology, is essential to support the fate of plant-microbe interactions. During evolution, plants have acquired the ability to perceive microbial molecules, initiating specific signal transduction cascades and leading to adapted response for microbe lifestyles (avirulent, virulent, or benefic). Plant survival will depend on the nature of the induced mechanisms. My PhD work, carried out on a simplified experimental system, contributes to the understanding of mechanisms underlying the determinism of plant-microbe interactions. We used Nicotiana tabacum cells in suspension exposed to microbial molecules derived from mutualistic or avirulent microbes. Using such a simplified system, we analyzed elements of the early signaling cascade and sugar fluxes. We have shown that CO4, which is originating from AMF, initiate early signaling components (rbohD-dependent H2O2, cytosolic Ca2+, MAPK activation) as cryptogein, a defense elicitor, but with distinct profile and amplitude. Those two molecules (CO4 and cryptogein) are responsible of different effects on sugar fluxes and the expression of the underlying sugar transporter genes. In addition, we presented an impact of the alteration of clathrin-mediated process on early signaling events triggered by cryptogein, as well as inward sugar fluxes and expression of sugar transporter genes. Finally, in silico analyses of sugar transporter interactome in Arabidopsis thaliana has provided some possible regulation mechanisms through the identification of new candidate proteins involved in sugar transporter regulation. These information open new perspectives towards a better understanding of the cellular and molecular mechanisms involved in plant-microbe interactions.

La distribution des sucres est un processus clé dans le développement de la plante et cours des interactions plantes/microorganismes.Une recherche des acteurs moléculaires impliqués dans la répartition des ressources carbonées au cours de l'interaction avec le champignon nécrotrophe B. cinerea a été réalisée. Plusieurs familles de transporteurs de sucres et d'invertases ont été ciblées, permettant d'établir une cartographie des gènes régulés transcriptionnellement lors de l'interaction. Le rôle de certains gènes candidats a été étudié par une approche de génomique fonctionnelle afin de mettre en évidence une fonction biologique de l'allocation du carbone dans la résistance de la plante aux champignons nécrotrophes. Un système d'interaction simplifié, basé sur un dialogue moléculaire sans contact physique entre une culture cellulaire d'A. thaliana et B. cinerea, a été développé. Il a permis de mesurer les flux de sucres ainsi que les activités enzymatiques et métaboliques pour chaque partenaire. Nos résultats montrent que B. cinerea entraine une forte augmentation de l'activité invertasique pariétale dans les tissus infectés, indiquant qu'une transition source/puits a lieu. Plusieurs transporteurs de sucres sont différentiellement exprimés, certains d'entre eux modulant le devenir de l'interaction. L'activité d'absorption d'hexoses et le métabolisme primaire des cellules hôtes sont fortement stimulés, démontrant l'importance de la compétition pour les sucres à l'interface plante/agent pathogène. En conclusion, l'absorption des sucres alimente le métabolisme énergétique des cellules hôtes et participe aux mécanismes de défense de la plante. / During plant development and upon pathogen infection, sugar allocation is a key process in plant physiology. Cell wall invertases and sugar transporters, involved in the sink strength, likely play a major role in the metabolic plant response. Molecular actors involved in carbohydrates allocation upon B. cinerea interaction have been identified using a transcriptional approach. Some gene families of sugar transporters and invertases have been targeted, allowing the establishment of a cartography of genes regulated during the interaction. To understand the biological role of carbon allocation during the interaction between plants and necrotrophic fungi, candidate genes have been studied using a functional genomics approach.A simplified interaction system has been developped, allowing a molecular dialogue between Arabidopsis and B. cinerea cells, without any physical contact. This system enables the monitoring of radiolabelled sugar uptake rates and some enzymatic and metabolomic activities for both the host cells and the pathogen, independently.Globally, our results demonstrate that B. cinerea infection leads to the transition from a source to a sink tissue, with a strong increase in cell wall invertase activity. The expression of some sugar transporter genes is also affected, while some of them (AtSTP1 and 13) are involved in the disease development. Besides the increase in hexose uptake activity, primary metabolism is deeply affected, highlighting the competition for apoplastic sugars that takes place at the plant/pathogen interface. Sugar retrieval appears to be a key process, fuelling host cells with energy and signal molecules, contributing to the plant defense mechanisms.