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La publication du premier séquençage d'un arbre - Le peuplier - et celle du champignon ectomycorhizien - Laccaria bicolor - a ouvert des nouvelles perspectives en biologie forestière. Les deux organismes constituent des modèles d'étude grâce à la connaissance de leur génome mais aussi de par leur importance économique, leur intérêt environnemental et leur capacité à s'associer de façon symbiotique. La symbiose mycorhizienne entre un champignon et une plante, améliore la nutrition minérale de la plante, et particulièrement sa nutrition phosphatée. La gestion de la forêt nécessite une meilleure compréhension des mécanismes qui servent à faire face à une faible fertilité en Pi dans le sol. Pour ce faire, nous avons réalisé la caractérisation des familles multigéniques des transporteurs de Pi de type PHT1 des deux organismes. Nous avons réussi à élucider les subtiles régulations de la famille de transporteurs de Pi entre ces deux partenaires, lors de différents états physiologiques de la plante et du champignon. Notre étude a mis en évidence l'effet positif observé sur la croissance des plantes lors de la mycorhization, celui-ci pouvant être étroitement lié à la régulation des certains transporteurs de Pi des deux partenaires (LbPht1;5 chez L.bicolor ; PtrPht1;9 et PtrPht1;12 chez le peuplier associé à L.bicolor et PtrPht1;10 associé à G.intraradices). D'autres transporteurs ont montré des réponses spécifiques à des stimuli comme la sénescence de feuilles (PtrPht1;1, PtrPht1;5, PtrPht1;6, PtrPht1;9 et PtrPht1;12), la carence en Pi (PtrPht1;9, PtrPht1;12 chez le peuplier, LbPht1;4 chez L. bicolor) et enfin nous avons pu observé les différents niveaux d'expression de chaque transporteur de Pi dans les différents compartiments tissulaires de chacun des partenaires. Cette mosaïque de réponses des membres de la famille PHT1 de ces deux organismes correspond à une première base de connaissances solide qui nous permettra de mieux comprendre les mécanismes du transport du Pi dans le sol des deux partenaires, par voie directe et symbiotique. / The publication of the first genome sequence of a tree - the poplar - and of an ectomycorrhizal fungus - Laccaria bicolor – has opened new prospects in forestry. Because their genomes are now known, and these species are important both economically and ecologically,Poplar and L. bicolor comprise an important model of plant : fungal interactions,. The mycorrhizal symbiosis, occurring between a fungus and a plant, improves plant mineral nutrition, especially for phosphorus. Forest management requires a better understanding of the mechanisms utilized by plants growing in low phosphorus soils. To increase our understanding of these processes, we characterized the multigenic families of Pi transporters (PHT1 type) of both organisms. We elucidated the subtle regulation of these transporters, from both partners, in various physiological states. Plants grew faster when mycorrhizal and this effect might be explained by the regulation of certain of the Pi transporters : LbPht1;5 in L.bicolor; PtrPht1;9 and PtrPht1;12 in poplar associated with L.bicolor, and PtrPht1;10 associated with G.intraradices. Other Pi transporters responded specifically to stimuli such as leaf senescence (PtrPht1;1, PtrPht1;5, PtrPht1;6, PtrPht1;9 and PtrPht1;12) or Pi starvation (PtrPht1;9, PtrPht1;12 in poplar, LbPht1;4 in L. bicolor). In addition, we observed different levels of expression of each Pi transporter in different tissues of each partner. This mosaic of responses of PHT1 family members provides an initial understanding of how mycorrhization affects Pi uptake from the fungus and plant. From this solid base of knowledge, the mechanisms by which each partner takes up Pi, either directly from soil, or indirectly via their symbiont, can be further expored.

Dada sua importância para o crescimento de diversas culturas, o fósforo encontra uso regular na produção de fertilizantes. Entretanto, o fato desse elemento apresentar-se na natureza em uma forma não assimilável pelos vegetais, fez com que a indústria de fertilizantes fosse forçada a desenvolver processos para sua dissolução. Esses processos costumam, em geral, ser agrupados segundo duas grandes rotas: a rota úmida e a rota térmica. Os processos que constituem a rota úmida caracterizam-se por promover a digestão do mineral de fósforo por meio de ácidos minerais tais como o sulfúrico e o fosfórico. Por conta desse tratamento são obtidos os superfosfatos simples e triplo. Já os processos compreendidos pela rota térmica valem-se, por sua parte, de energia térmica para modificar a estrutura da rocha fosfatada, a partir do que serão gerados os chamados termofosfatos. As rotas úmida e térmica já foram comparadas em diversas ocasiões, em geral segundo critérios de cunho técnico e econômico. O presente trabalho realiza também a referida comparação, agora porém sob o ponto de vista de desempenho ambiental, empregando, para tanto, a técnica de Análise de Ciclo de Vida (ACV). Selecionou-se como representante da rota úmida o superfosfato triplo (TSP), enquanto que o termofosfato magnesiano fundido (FMP) foi usado para retratar a rota térmica. Para efeito da execução de ambos os estudos de ACV, tomou-se por função para o TSP e para o FMP, a capacidade dessesprodutos em fornecer fósforo às plantas. No que se refere à definição dos sistemas de produto, decidiu-se adotar uma abordagem do tipo \"cradle to gate\" para a aplicação da ACV. Em razão desse fato, o sistema de produto de TSP passou a contemplar as etapas de lavra e concentração da rocha fosfática, as produções de ácido sulfúrico e ácido fosfórico e a manufatura em si desse fertilizante fosfatado. ) Os subsistemas auxiliares de transporte, e das gerações de energia elétrica e de gás natural foram também considerados no estabelecimento do referido sistema de produto. O sistema de produto do FMP foi estruturado segundo a mesma abordagem, passando por conta disso a ser composto pelos subsistemas de lavra e de britagem da rocha fosfática, pelo transporte de serpentinito, pelas gerações de energia elétrica e de gás natural, e pela produção em si do fertilizante. Por fim, para que fosse possível fornecer um quadro preciso do desempenho ambiental dos sistemas sob avaliação, houve-se por bem selecionar as categorias de impacto de Aquecimento Global (PAG), Depleção de Camada de Ozônio (PDCO), Toxicidade Humana (PTH), Ecotoxicidade Aquática (PEa), Acidificação (PAc) e Eutrofização (PEu). A realização de estudos de ACV segundo essas e outras premissas mostrou que o desempenho do sistema de produto de TSP foi superior ao do FMP para todas as categorias de impacto selecionadas, excetuando-se os potenciais de Acidificação e de Eutrofização. Em termos do Potencial de AquecimentoGlobal, o FMP mostrou-se quatro vezes mais agressivo ao meio ambiente que seu homólogo fosfatado. Esse quadro se repete também para a Ecotoxicidade Aquática; desde o ponto de vista dessa categoria de impactos, a produção de TSP traz efeitos lesivos sobre o meio ambiente com uma magnitude 3,3 vezes menor que se o mesmo elemento fosse disponibilizado como FMP. Por outro lado, no que se refere à Eutrofização, por meio da qual são de fato medidos os impactos ambientais gerados pelas perdas de fósforo ao longo de ambos os processamentos, o FMP apresentou melhor desempenho ambiental que o TSP. / Given its importance to the growth of several crops, the element phosphorous is regularly used for the production of fertilizers. Nevertheless, the fact that this element is found in Nature in a non-assimilable form has forced the fertilizer industry to develop processes for its dissolution. These processes are usually divided in two major groups: the wet route and the thermal route. The processes that constitute the wet route characteristically digest the phosphate rock using mineral acids such as the sulfuric acid and the phosphoric acid. In this treatment are produced, respectively, the single superphosphate and the triple superphosphate. On the other hand, the processes belonging to the thermal route use thermal energy to modify the structure of the phosphate rock generating the so-called thermophosphates. The wet and thermal routes have been compared in several occasions, usually according to technical and economic criteria. This study also performs such a comparison, but now on an environmental performance approach, using for this the Life Cycle Assessment methodology (LCA). The triple superphosphate (TSP) was chosen to represent the wet route whereas the fused magnesium thermophosphate (FMP) was chosen to reflex the thermal route. In both LCA studies the function of the products was deemed to be to supply phosphorous to crops. The product systems were defined using a \"cradle-to-gate\" approach. Therefore the TSP product system contemplates:the mining and concentration of the phosphate rock, the production of sulfuric and phosphoric acids and the TSP manufacture itself. The auxiliary subsystems of transportation and the electricity and natural gas generation subsystems were also considered in the above-mentioned system. ) The FMP product system was structured in a similar way and contemplates: the mining and crushing of the phosphate rock, the serpentinite transportation, the electricity and natural gas generation and the FMP manufacture itself. Finally, in order to give a picture as accurate as possible of the environmental performance of the systems under evaluation, the following impact categories were chosen: Global Warming (GWP), Ozone Layer Depletion (OLDP), Human Toxicity (HTP), Aquatic Ecotoxicity (AETP), Acidification (AP) and Eutrophication (EP). The LCA studies have shown that the TSP product system performed better than the FMP product system in all the selected impact categories but Acidification and Eutrophication. In terms of Global Warming Potential, the FMP was shown to be four times more aggressive to the environment than TSP. This scenario repeats itself also for Aquacic Ecotoxicity. In this category, the TSP has an effect 3.3 times lower than the effect of the FMP. On the other hand, regarding Eutrophication, through which are in fact measured the environmental impacts generated by phosphorous losses during both product systems, FMP has presented a better performance.

Dada sua importância para o crescimento de diversas culturas, o fósforo encontra uso regular na produção de fertilizantes. Entretanto, o fato desse elemento apresentar-se na natureza em uma forma não assimilável pelos vegetais, fez com que a indústria de fertilizantes fosse forçada a desenvolver processos para sua dissolução. Esses processos costumam, em geral, ser agrupados segundo duas grandes rotas: a rota úmida e a rota térmica. Os processos que constituem a rota úmida caracterizam-se por promover a digestão do mineral de fósforo por meio de ácidos minerais tais como o sulfúrico e o fosfórico. Por conta desse tratamento são obtidos os superfosfatos simples e triplo. Já os processos compreendidos pela rota térmica valem-se, por sua parte, de energia térmica para modificar a estrutura da rocha fosfatada, a partir do que serão gerados os chamados termofosfatos. As rotas úmida e térmica já foram comparadas em diversas ocasiões, em geral segundo critérios de cunho técnico e econômico. O presente trabalho realiza também a referida comparação, agora porém sob o ponto de vista de desempenho ambiental, empregando, para tanto, a técnica de Análise de Ciclo de Vida (ACV). Selecionou-se como representante da rota úmida o superfosfato triplo (TSP), enquanto que o termofosfato magnesiano fundido (FMP) foi usado para retratar a rota térmica. Para efeito da execução de ambos os estudos de ACV, tomou-se por função para o TSP e para o FMP, a capacidade dessesprodutos em fornecer fósforo às plantas. No que se refere à definição dos sistemas de produto, decidiu-se adotar uma abordagem do tipo \"cradle to gate\" para a aplicação da ACV. Em razão desse fato, o sistema de produto de TSP passou a contemplar as etapas de lavra e concentração da rocha fosfática, as produções de ácido sulfúrico e ácido fosfórico e a manufatura em si desse fertilizante fosfatado. ) Os subsistemas auxiliares de transporte, e das gerações de energia elétrica e de gás natural foram também considerados no estabelecimento do referido sistema de produto. O sistema de produto do FMP foi estruturado segundo a mesma abordagem, passando por conta disso a ser composto pelos subsistemas de lavra e de britagem da rocha fosfática, pelo transporte de serpentinito, pelas gerações de energia elétrica e de gás natural, e pela produção em si do fertilizante. Por fim, para que fosse possível fornecer um quadro preciso do desempenho ambiental dos sistemas sob avaliação, houve-se por bem selecionar as categorias de impacto de Aquecimento Global (PAG), Depleção de Camada de Ozônio (PDCO), Toxicidade Humana (PTH), Ecotoxicidade Aquática (PEa), Acidificação (PAc) e Eutrofização (PEu). A realização de estudos de ACV segundo essas e outras premissas mostrou que o desempenho do sistema de produto de TSP foi superior ao do FMP para todas as categorias de impacto selecionadas, excetuando-se os potenciais de Acidificação e de Eutrofização. Em termos do Potencial de AquecimentoGlobal, o FMP mostrou-se quatro vezes mais agressivo ao meio ambiente que seu homólogo fosfatado. Esse quadro se repete também para a Ecotoxicidade Aquática; desde o ponto de vista dessa categoria de impactos, a produção de TSP traz efeitos lesivos sobre o meio ambiente com uma magnitude 3,3 vezes menor que se o mesmo elemento fosse disponibilizado como FMP. Por outro lado, no que se refere à Eutrofização, por meio da qual são de fato medidos os impactos ambientais gerados pelas perdas de fósforo ao longo de ambos os processamentos, o FMP apresentou melhor desempenho ambiental que o TSP. / Given its importance to the growth of several crops, the element phosphorous is regularly used for the production of fertilizers. Nevertheless, the fact that this element is found in Nature in a non-assimilable form has forced the fertilizer industry to develop processes for its dissolution. These processes are usually divided in two major groups: the wet route and the thermal route. The processes that constitute the wet route characteristically digest the phosphate rock using mineral acids such as the sulfuric acid and the phosphoric acid. In this treatment are produced, respectively, the single superphosphate and the triple superphosphate. On the other hand, the processes belonging to the thermal route use thermal energy to modify the structure of the phosphate rock generating the so-called thermophosphates. The wet and thermal routes have been compared in several occasions, usually according to technical and economic criteria. This study also performs such a comparison, but now on an environmental performance approach, using for this the Life Cycle Assessment methodology (LCA). The triple superphosphate (TSP) was chosen to represent the wet route whereas the fused magnesium thermophosphate (FMP) was chosen to reflex the thermal route. In both LCA studies the function of the products was deemed to be to supply phosphorous to crops. The product systems were defined using a \"cradle-to-gate\" approach. Therefore the TSP product system contemplates:the mining and concentration of the phosphate rock, the production of sulfuric and phosphoric acids and the TSP manufacture itself. The auxiliary subsystems of transportation and the electricity and natural gas generation subsystems were also considered in the above-mentioned system. ) The FMP product system was structured in a similar way and contemplates: the mining and crushing of the phosphate rock, the serpentinite transportation, the electricity and natural gas generation and the FMP manufacture itself. Finally, in order to give a picture as accurate as possible of the environmental performance of the systems under evaluation, the following impact categories were chosen: Global Warming (GWP), Ozone Layer Depletion (OLDP), Human Toxicity (HTP), Aquatic Ecotoxicity (AETP), Acidification (AP) and Eutrophication (EP). The LCA studies have shown that the TSP product system performed better than the FMP product system in all the selected impact categories but Acidification and Eutrophication. In terms of Global Warming Potential, the FMP was shown to be four times more aggressive to the environment than TSP. This scenario repeats itself also for Aquacic Ecotoxicity. In this category, the TSP has an effect 3.3 times lower than the effect of the FMP. On the other hand, regarding Eutrophication, through which are in fact measured the environmental impacts generated by phosphorous losses during both product systems, FMP has presented a better performance.

Ce mémoire présente l’étude de protections anti-oxydation (PAO) de matériaux composites SiC/SiC. Ces matériaux sont dotés de propriétés spécifiques (légèreté, résistance aux chocs thermiques et tenue mécanique à haute température) par l’association de fibres et d’une matrice SiC liées par une interphase en carbone. Néanmoins, cette interphase s’oxyde dès 400°C entraînant la décohésion entre les fibres et la matrice et ainsi la perte des propriétés mécaniques. En vue d’applications dans le domaine de l’aéronautique civile, la PAO doit être opérationnelle à basse température (400-650°C) et sur de longues périodes (de l’ordre de plusieurs milliers d’heures). Deux types de protections ont été proposés et étudiés: (i) une protection rigide formée par traitement thermique d’un mélange entre une solution de phosphate d’aluminium et de silice. La structure du revêtement a ensuite été caractérisée par RMN, DRX et MEB et mise en relation avec l’efficacité de la protection. (ii) une protection dynamique basée sur l’utilisation de verres de phosphate formulés afin de présenter des caractéristiques de viscosité optimales pour la PAO. Les formulations ont été caractérisées par analyses thermiques (ATD, ATM, HSM) et par RMN. Une démarche raisonnée a été mise en œuvre en corrélant les propriétés thermiques aux caractéristiques structurales des oxydes utilisés. Ces deux revêtements étudiés améliorent la durée de vie du matériau, mais sans atteindre la spécification attendue. Nous concluons que la protection dynamique est plus adaptée à l’application basse température et proposons des voies d’amélioration. / This work presents anti-oxidation coating (AOC) of composite materials SiC/SiC. These materials have specific properties (lightness, excellent resistance to thermal shocks, and mechanical endurance at very high temperature) due to the combination of SiC fibers and SiC matrix linked by a C interphase. This interphase oxidizes under air above 400°C, leading to fiber/matrix debonding and thus compromising the mechanical properties. In view of application in civil aeronautics, the AOC have to be operational at low temperatures (400-650°C) and for prolonged periods (of the order of thousands of hours). Two types of coatings were proposed and examined: (i) a rigid coating obtained through the thermal treatment of a mixture of aluminium phosphate solution and silica. The structure of the coating has been characterized by NMR, XRD and SEM and related to the efficiency of the protection. (ii) a dynamic coating, based on phosphate glasses formulated to present optimum viscosity characteristics. The different glasses were characterized by thermal analyses (DTA, TMA, HSM) and by NMR. A thought approach was implemented by correlation between thermal properties and structural characteristics of oxides used. These two coatings improve the material lifetime even if the expected specification was not accomplished. To conclude, the dynamic coating is more adapted to low temperature use and we suggest some improvements.

To ensure a continuous supply of phosphate, living organisms have devised complex mechanisms to regulate the uptake and subsequent utilization of this essential nutrient. An important aspect of phosphate metabolism is the storage of excess phosphate as the polymer, polyphosphate. Despite the importance of this polymer to all living organisms, much needs to be learned about its synthesis, storage, or utilization. Furthermore, little is known about the regulatory mechanisms that determine when polyphosphate synthesis or degradation is appropriate. Our work has shown that polyphosphate is a dynamic molecule whose levels fluctuate during the cell cycle. Polyphosphate levels are high in G1, and subsequently drop as the cell uses free phosphate during cell division. Mitotic induction of phosphate regulatory genes, including the acid phosphatase gene PHO5, replenishes polyphosphate levels late in the mitosis. Furthermore, we have shown that Mcm1 and Fkh1, two cell cycle dependent transcriptional activators, contribute to mitotic activation of PHO5. In addition, we have elucidated the importance of regulating polyphosphate synthesis. Strains lacking the cyclin Pho80 have increased expression of the polyphosphate synthase genes, PHM1-4, and thus have highly elevated polyphosphate levels. Hyperaccumulation of polyphosphate results in severe growth defects on medium containing high levels of sorbitol, presumably through the polyphosphate-dependent overacidification of the vacuole.

Glycerophosphodiesters are products of phospholipase B-mediated hydrolysis of phospholipids. Their transport and metabolism is elaborated in the fungal pathogen, Candida albicans, as compared to the non-pathogenic Saccharomyces cerevisiae. C. albicans contains four ORFs (CaGIT1-4) predicted to encode transporters for glycerophosphodiesters, compared to one (ScGIT1) in S. cerevisiae. Here I have identified the gene products responsible for glycerophosphoinositol (GroPIns) and glycerophosphocholine (GroPCho) transport. C. albicans strain lacking ORF 19.34, which codes for CaGit1, is unable to transport intact GroPIns. Transport activity can be rescued by reintegration of one copy of CaGIT1 back into the genome. Similarly, a strain lacking CaGIT3 (ORF 19.1979) and CaGIT4 (ORF 19.1980) is unable to transport intact GroPCho into the cell. Reintegrating one copy of either CaGIT3 or CaGIT4 can rescue GroPCho transport activity. Initial transport assays and kinetic analyses indicate that CaGit3 is responsible for the majority of GroPCho transport activity. In addition, I present evidence that CaGDE1 (ORF 19.3936) codes for an enzyme with glycerophosphodiesterase activity against GroPCho. Homozygous deletion of CaGDE1 results in a buildup of internal GroPCho, which is restored to wild type accumulation by reintegration of one copy of CaGDE1 into the genome. The transcriptional regulator, CaPho4, was shown to positively regulate the expression of CaGIT1, CaGIT3, CaGIT4, and CaGDE1. Finally, glycerophosphodiester transport and metabolism was active under physiological relevant conditions that C. albicans may experience in the human host. / Bayer School of Natural and Environmental Sciences; / Biological Sciences / PhD; / Dissertation;

Extraction with sodium phosphate has been used as a method of accessing arsenic in soils. Arsenic extraction efficiency by phosphate from rice-paddy soils of Bangladesh usually has been low and highly variable between soils. The major objectives of this study were to examine the relationships between phosphate-extractable arsenic and soil iron-oxide composition and to investigate the experimental factors which might influence arsenic-extraction efficiency from rice-paddy soils of Bangladesh by phosphate. Statistical analysis of approximately 500 surface soils from Bangladesh indicated that phosphate-extractable arsenic was well correlated with total soil arsenic (r2 = 0.832) and oxalate-extractable arsenic (r2 = 0.825), though extraction efficiency varied widely (5 - 54 % of the total soil arsenic). The thanas with the lowest arsenic contents generally also had the soils with the lowest arsenic-extraction efficiencies. Quantity of phosphate-extractable arsenic was weakly correlated with the soil iron-oxide content, but extraction efficiency (i.e., the proportion of phosphate-extractable arsenic to total soil arsenic) was not correlated with any iron-oxide parameter. Arsenic extraction was strongly influenced by reaction variables such as sample grinding, phosphate concentration, principal counterion, reaction pH, and reaction time. The extraction efficiency was impacted by the influence of these individual factors on reaction kinetics and accessibility of arsenic adsorption sites for ligand exchange by phosphate. A portion of the arsenic was readily exchanged during the first few hours of extraction, followed by a much slower subsequent extraction. These results indicate that some of the arsenic is easily exchanged, but for a substantial portion of the arsenic, either the reaction kinetics is very slow or the sites are not accessible for reaction with phosphate. Extraction by phosphate is a useful procedure for the assessment of readily ligand-exchanged arsenic.

To ensure a continuous supply of phosphate, living organisms have devised complex mechanisms to regulate the uptake and subsequent utilization of this essential nutrient. An important aspect of phosphate metabolism is the storage of excess phosphate as the polymer, polyphosphate. Despite the importance of this polymer to all living organisms, much needs to be learned about its synthesis, storage, or utilization. Furthermore, little is known about the regulatory mechanisms that determine when polyphosphate synthesis or degradation is appropriate. Our work has shown that polyphosphate is a dynamic molecule whose levels fluctuate during the cell cycle. Polyphosphate levels are high in G1, and subsequently drop as the cell uses free phosphate during cell division. Mitotic induction of phosphate regulatory genes, including the acid phosphatase gene PHO5, replenishes polyphosphate levels late in the mitosis. Furthermore, we have shown that Mcm1 and Fkh1, two cell cycle dependent transcriptional activators, contribute to mitotic activation of PHO5. In addition, we have elucidated the importance of regulating polyphosphate synthesis. Strains lacking the cyclin Pho80 have increased expression of the polyphosphate synthase genes, PHM1-4, and thus have highly elevated polyphosphate levels. Hyperaccumulation of polyphosphate results in severe growth defects on medium containing high levels of sorbitol, presumably through the polyphosphate-dependent overacidification of the vacuole.