Dans les conditions naturelles, les plantes sont soumises à des conditions environnementales très variées qui peuvent conduire à un excès d'énergie dans les chloroplastes, résultant en une production d'espèces réactives de l'oxygène (ERA). Pour faire face à ces ERA, les plantes ont développé différents mécanismes de protection, comme des alkénal réductases, des peroxyrédoxines et des lipocalines. Le travail présenté dans cette thèse a pour but de caractériser et de déterminer leur importance dans la protection des contre les stress oxydants. Une analyse préliminaire de mutants d'Arabidopsis a mis en évidence le rôle important des peroxyrédoxines à 2 cystéines et des lipocalines dans la tolérance au stress photooxydant. Cette thèse s'est surtout concentrée sur les lipocalines. Deux lipocalines ont été récemment identifiées chez les plantes, TIL (lipocaline thermoinduite) et CHL (chloroplastique), toutes les deux induites par des conditions de stress. Chez Arabidopsis, chaque lipocaline semble être spécialisée dans la réponse à des conditions différentes: chaleur (AtTIL) et fortes lumières (AtCHL). Le double mutant AtCHL KO × AtTIL KO est plus sensible à la chaleur et la forte lumière que les simples mutants. La mutation de AtCHL a augmenté fortement la photosensibilité de mutants (vte1, npq1) affectés dans des mécanismes de protection des lipides (tocopherols, zéaxanthine), confirmant ainsi le rôle des lipocalines dans la protection contre la peroxydation lipidique. Les résultats obtenus dans cette thèse montrent que les lipocalines AtTIL and AtCHL ont des fonctions redondantes dans la protection des lipides qui sont essentielles à la résistance des plantes au stress. / Under field conditions, plants are exposed to various environmental conditions that can lead to an excess of energy in the chloroplasts, resulting in the production of toxic reactive oxygen species (ROS). To cope with the harmful effects of ROS, plants have developed various protection mechanisms, such as alkenal-reductases, peroxiredoxins and lipocalins. The work performed in this thesis aimed at understanding their importance in the protection against lipid peroxidation. A first screening of Arabidopsis mutants lacking one of those mechanisms brought into light that 2-Cys PRX and lipocalins are important for the tolerance against photooxidative stress. This thesis is focused mainly on lipocalins, a group of proteins recognized as carriers of small lipophilic molecules. However, two true lipocalins have been recently identified in plants, the temperature-induced lipocalin (TIL) and the chloroplastic lipocalin (CHL), their expression beeing induced by various abiotic stresses. Each lipocalin appeared to be specialized in the responses to specific stress conditions in Arabidopsis, with AtTIL and AtCHL playing a protective role against heat and high light, respectively. The double mutant AtCHL KO × AtTIL KO deficient in both lipocalins was more sensitive to temperature, drought and light stresses than the single mutants. Seeds of the AtCHL KO × AtTIL KO double mutant were very sensitive to natural and artificial aging, and again this phenomenon was associated with the oxidation of polyunsaturated lipids. The results obtained in this thesis show that AtTIL and AtCHL have overlapping functions in lipid protection which are essential for stress resistance and survival.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / In this study were characterized biochemical and physiological mechanisms arising from photoinhibition that act in the modulation of photochemical activity and oxidative protection, helping to reduce photo oxidative damage in response to drought and high light stresses in cashew plants. The seedlings were obtained from seeds and grown in substrates by mixing sand and vermiculite (ratio 1:1) in plastic bags, with a volume of 2 liters. Drought stress was imposed by withholding the water supply to the plants ( 20 days) in greenhouse conditions. For exposure to light treatments and recovery to the light effect, the plants were placed in a chamber with controlled conditions of relative humidity (60% Â5) and temperature (30ÂC Â2). The plants subjected to drought stress showed a marked decrease in photosynthesis when exposed to excess light (combination of drought with high luminosity), compared with those irrigated. This higher sensitivity of CO2 fixation to excess light on plants under drought was associated with a higher intensity of damage to the photochemical apparatus, as indicated by measures of effective quantum efficiency and apparent electron transport rate. The data show that the reduction of photosynthetic activity in response to excess light, has been attributed in part to process photoinhibition occurred under these conditions as shown by a drastic reduction in potential maximum quantum efficiency of PSII (given by Fv/Fm ratio). The results also suggest that the intense photoinhibition, triggered by excess light, may have assisted in photoprotection, because under these conditions there was no oxidative damage, as indicated by the absence of changes in H2O2 content and lipid peroxidation (TBARS content). This photochemistry protection, assigned to photoinhibition in this study, is enhanced due to non-occurrence of dissipation of excess energy through non-photochemical quenching (NPQ), because this mechanism did not show significant changes after long periods of exposure to excessive light. However, the energy dissipation through NPQ in response to short time of exposure to excessive light, observed here indicates that when the PSII activity is normal this mechanism is essential to the protection photochemistry. The role of photoinhibition for photo oxidative protection can be attributed to the effect of this process on reducing the activity of PSII and consequent lower photochemical activity, resulting in lower electron transfer. This suggestion is also reiterated in this study based on the reduction of the content of D1 protein, a major component of structural and functional PSII, and plastocyanin (PC), another major carrier of electrons. The modulation these proteins content may have contributed to restrict the formation of ROS and consequent oxidative damage under conditions inducing photo oxidatives stress such as drought associated with high luminosities, in this species. / No presente estudo foram caracterizados mecanismos bioquÃmicos e fisiolÃgicos decorrentes da fotoinibiÃÃo e que atuam na modulaÃÃo da atividade fotoquÃmica e da proteÃÃo oxidativa, auxiliando na reduÃÃo de danos fotoxidativos, em reposta aos estresses de seca e luminosidade elevada em cajueiro. As mudas foram obtidas a partir de sementes e cultivadas em substrato composto pela mistura de areia e vermiculita (proporÃÃo 1:1) em sacos plÃsticos, com volume de 2 L. O estresse hÃdrico foi aplicado pela suspenÃÃo da rega (Â20 dias) em condiÃÃes de casa de vegetaÃÃo. Para a exposiÃÃo aos tratamentos de luminosidade e de recuperaÃÃo ao efeito da luz, as plantas foram acondicionadas em cÃmara com condiÃÃes controladas de umidade relativa (60% Â5) e temperatura (30ÂC Â2). As plantas submetidas ao estresse hÃdrico apresentaram uma intensa reduÃÃo da fotossÃntese quando expostas ao excesso de luz (combinaÃÃo de seca com luminosidade elevada), se comparadas com aquelas irrigadas. Essa maior sensibilidade da fixaÃÃo de CO2 ao excesso de luz nas plantas sob seca foi relacionada com uma maior intensidade de danos no aparato fotoquÃmico, conforme indicado pelas medidas de eficiÃncia quÃntica efetiva e taxa aparente de transporte de elÃtrons. A reduÃÃo da atividade fotossintÃtica, em resposta ao excesso de luz, foi atribuÃda em parte ao processo de fotoinibiÃÃo ocorrido nessas condiÃÃes, conforme demonstrado pela drÃstica reduÃÃo da eficiÃncia quÃntica potencial mÃxima do PSII (dada pela relaÃÃo Fv/Fm). Os resultados sugerem tambÃm que a intensa fotoinibiÃÃo, desencadeada pelo excesso de luz, pode ter auxiliado na fotoproteÃÃo, pois nessas condiÃÃes nÃo ocorreu danos oxidativos, conforme indicado pela ausÃncia de mudanÃas no conteÃdo de H2O2 e na peroxidaÃÃo lipÃdica (conteÃdo de TBARS). Essa proteÃÃo fotoquÃmica, atribuÃda a fotoinibiÃÃo nesse estudo, à reforÃada devido a nÃo ocorrÃncia de dissipaÃÃo do excesso de energia por meio do quenching nÃo fotoquÃmico (NPQ), pois esse mecanismo nÃo apresentou mudanÃas significativas apÃs longos perÃodos de exposiÃÃo ao excesso de luz. No entanto, a dissipaÃÃo de energia por meio do NPQ em respostas a curtos perÃodos de exposiÃÃo ao excesso de luz, aqui observado, indica que quando a atividade do PSII està normal esse mecanismo à essencial para a proteÃÃo fotoquÃmica. O papel da fotoinibiÃÃo para a proteÃÃo fotoxidativa pode ser atribuido ao efeito desse processo na reduÃÃo da atividade dos PSII e consequente menor atividade fotoquÃmica, o que resultaria na menor transferÃncia de elÃtrons. Essa sugestÃo à ainda reinterada no presente estudo com base na reduÃÃo do conteÃdo da proteina D1, um dos principais componentes estrutural e funcional do PSII, e da plastocianina (PC), outro importante carreador de elÃtrons. A modulaÃÃo do conteÃdo dessas proteÃnas pode ter contribuÃdo para restringir a formaÃÃo de EROs e consequente danos oxidativos sob condiÃÃes indutoras de estresse fotoxidativos, como a seca associada com luminosidades elevadas, nessa espÃcie.