This thesis focuses on the nature of heme metabolism in various eukaryotes. One of the aims was the elucidation of the origin of the unique heme biosynthesis pathway in apicomplexan parasites through a comparative study of their photosynthetic relative Chromera velia combining molecular biology, biochemistry and bioinformatics approach. Using similar strategy, I have also investigated the origin and spatial organization of tetrapyrrole biosynthesis in Euglena gracilis. Based on the phylogenetic data I described the complex evolution of heme metabolism in kinetoplastid flagellates including pathogenic trypanosomes. I revealed that one of them (Phytomonas) does not require heme for viability by the combination of various biochemical and molecular biology experiments and bioinformatic analyses.

Les plastes primaires proviennent d'une cyanobactérie qui a établi une relationendosymbiotique avec un hôte eucaryote. Cet événement a donné naissance au super-groupeArchaeplastida qui inclut les Viridiplantae (algues vertes et plantes terrestres), les Rhodophyta (alguesrouges) et les Glaucophyta. Suite à l'endosymbiose primaire, les algues rouges et vertes ont étendu lacapacité de photosynthèse à d'autres lignées eucaryotes via des endosymbioses secondaires. Bien quedes progrès considérables aient été réalisés dans la compréhension de l'évolution des eucaryotesphotosynthétiques, d'importantes questions sont restées ouvertes, telles que l’identité de la lignéecyanobactérienne la plus proche des plastes primaires ainsi que le nombre et l'identité des partenairesdans les endosymbioses secondaires.Ma thèse a consisté à étudier l'origine et l'évolution précoce des eucaryotes photosynthétiques enutilisant des approches phylogénétiques et phylogénomiques. Je montre par mon travail que les plastesprimaires ont évolué à partir d'un symbiote phylogénétiquement proche de Gloeomargarita lithophora,une cyanobactérie représentant un clade s’étant diversifié précocement et qui a été détectéeuniquement dans les milieux terrestres. Ce résultat fournit des pistes nouvelles sur le contexteécologique dans lequel l'endosymbiose primaire a probablement eu lieu. En ce qui concerne l'évolutiondes lignées eucaryotes avec des plastes secondaires, je montre que les génomes nucléaires deschlorarachniophytes et des euglénophytes, deux lignées photosynthétiques avec des plastes dérivésd'algues vertes, encodent un grand nombre de gènes acquis par transferts depuis des algues rouges.Enfin, je mets en évidence que SELMA, la machinerie de translocation des protéines à travers laseconde membrane externe des plastes rouges secondaires à quatre membranes, a une histoireétonnamment compliquée aux implications évolutives importantes : les cryptophytes ont recruté unensemble de composants de SELMA différent de ceux des haptophytes, straménopiles et alvéolés.Ainsi, ma thèse a permis d’identifier pour la première fois la lignée cyanobactérienne la plus proche desplastes primaires et apporte de nouvelles pistes pour éclaircir les événements complexes qui ontjalonné l’évolution des eucaryotes photosynthétiques secondaires. / Primary plastids derive from a cyanobacterium that entered into an endosymbioticrelationship with a eukaryotic host. This event gave rise to the supergroup Archaeplastida whichcomprises Viridiplantae (green algae and land plants), Rhodophyta (red algae) and Glaucophyta. Afterprimary endosymbiosis, red and green algae spread the ability to photosynthesize to other eukaryoticlineages via secondary endosymbioses. Although considerable progress has been made in theunderstanding of the evolution of photosynthetic eukaryotes, important questions remained debatedsuch as the present-day closest cyanobacterial lineage to primary plastids as well as the number andidentity of partners in secondary endosymbioses.The main objectives of my PhD were to study the origin and evolution of plastid-bearing eukaryotesusing phylogenetic and phylogenomic approaches to shed some light on how primary and secondaryendosymbioses occurred. In this work, I show that primary plastids evolved from a close relative ofGloeomargarita lithophora, a recently sequenced early-branching cyanobacterium that has been onlydetected in terrestrial environments. This result provide interesting hints on the ecological setting whereprimary endosymbiosis likely took place. Regarding the evolution of eukaryotic lineages with secondaryplastids, I show that the nuclear genomes of chlorarachniophytes and euglenids, two photosyntheticlineages with green alga-derived plastids, encode for a large number of genes acquired by transfersfrom red algae. Finally, I highlight that SELMA, the translocation machinery putatively used to importproteins across the second outermost membrane of secondary red plastids with four membranes, has asurprisingly complex history with strong evolutionary implications: cryptophytes have recruited a set ofSELMA components different from those present in haptophytes, stramenopiles and alveolates.In conclusion, during my PhD I identified for the first time the closest living cyanobacterium to primaryplastids and provided new insights on the complex evolution that have undergone secondary plastid-bearing eukaryotes

Made available in DSpace on 2015-03-26T13:30:30Z (GMT). No. of bitstreams: 1 texto completo.pdf: 4228390 bytes, checksum: af8843c122dbdf2a43677202853bea97 (MD5) Previous issue date: 2010-02-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Interactions between arthropods and microorganisms have been receiving more attention due to the impact that these symbiotic associations causes on their macrosymbionts. Reproduction is the main biological parameters affected with cytoplasmic incompatibility and thelytokous parthenogenesis; however, endosymbionts can help arthropods to exploit host plants. In the feeding time, the herbivore causes injuries in plant tissue and induces a defense response in the plant that respond with production and posterior release of a variety of chemical compounds. However, in one case, plant defense system is affected and disturbed bay herbivore mite. It was detected in tomato plants when the mite Tetranychus evansi Baker and Pritchard interfered in the defense system of these plants, making them more nutritionally adequate for development of this mite. Continuing in the research line, the actual study locked for a better understanding of the interaction tomato and the mite T. evansi, in the possible symbiotic association presence between mite and microorganism that could promote advantages for this mite overcome the tomato defenses. Initially, molecular and microbiological analyses were performed to verify the presence of microorganisms. Later, mites were treated with the antibiotic tetracycline hydrochloride 0.2% that was the treated population. Control was represented from an untreated population and the possible advantage given by microorganism presence in the interior of the T. evansi organism was evaluated by the oviposition performance of the mite populations. Mite egg number was checked daily for four days. As substrate were used tomato leaves from Santa Clara I-5300 variety. In another experiment, chapter two, was evaluated the levels of proteinase inhibitors and total aldehydes that are present tomato damaged by treated and untreated mite population. As result from a first chapter, molecular analysis, using the method of polymerase chain reaction (PCR) confirmed the presence of bacterial DNA fragments in the untreated population and DNA fragments absence in the treated population. These results were confirmed by the microbiological analysis.Accessing the biological performance of treated and untreated was obtained that oviposition rate from treated population was 2.8 times lower than that of untreated population. In the second chapter was observed that the level of total aldehydes was low in plants damaged for treated and untreated treatments. These plants where put as feed and oviposition substrate for treated and untreated populations. Treated population put a same egg number on plants previously damaged by both populations. Already, untreated population showed highest oviposition rate on plants previously damaged by treated population. Thus, we can resume that the presence of microorganisms may be associated with nutritional benefits and detoxification. In determining the levels of proteinase inhibitors, treatment consists of plants infested with mites treated showed significantly lower value compared to other treatments. And the occurrence of microorganism and mite interaction can be related to potential of T. evansi to explore the nutritional quality and to confound the defense system of tomato plants. Future researches need be carried for to identify the microorganisms involved in the association and to study other possible effects of this interaction on other biological parameters of T. evansi. / Interações entre artrópodes e microrganismos vêm recebendo maior atenção devido ao impacto que essas associações simbióticas promovem nos seus macrosimbiontes. A reprodução é um dos parâmetros biológicos afetados com a incompatibilidade citoplasmática e a partenogênese telítoca, contudo, os endossimbiontes auxiliam os artrópodes a explorar plantas hospedeiras. No momento da alimentação, o herbívoro provoca injúrias no tecido vegetal e induz uma resposta de defesa da planta com produção e/ou liberação de uma diversidade de compostos químicos. O ácaro Tetranychus evansi Baker e Pritchard consegue interferir no sistema de defesa de suas plantas hospedeiras, tornando-as nutricionalmente mais adequadas para o seu desenvolvimento. Nesse contexto, este trabalho procurou compreender melhor a interação tomateiro-ácaro T. evansi, além da possível associação simbiótica promovendo vantagens para esse ácaro superar as defesas do seu hospedeiro. Para isto, os ácaros foram submetidos ao tratamento com o antibiótico hidroclorido de tetraciclina 0,2 %. Assim foram utilizados para os ensaios ácaros tratados com antibiótico e ácaros não tratados. Análises moleculares e microbiológicas foram realizadas para verificar a presença dos microrganismos. A oviposição foi avaliada diariamente durante quatro dias, utilizando folhas de tomate da variedade Santa Clara I-5300 como substrato. Os níveis de inibidores de proteinases e aldeídos totais foram determinados em tomateiros. As plantas aos 45 dias de idade foram infestadas com o ácaro T. evansi tratado com hidroclorido de tetraciclina 0,2 %, T. evansi não tratado e T. urticae não tratado, e como controle foram utilizadas plantas não infestadas. Plantas sob os mesmos tratamentos foram utilizadas como substrato para a avaliação da oviposição do T. evansi tratado com tetraciclina e não tratado. A análise molecular, utilizando o método da reação em cadeia da polimerase (PCR), confirmou a presença de fragmentos DNA bacteriano nos ácaros não tratados e ausência nos ácaros tratados com tetraciclina. As análises microbiológicas confirmaram os resultados obtidos por PCR. A taxa de oviposição dos ácaros tratados foi 2,8 vezes inferior a dos ácaros não tratados. A presença dos microrganismos pode estar associada a benefícios nutricionais e desintoxicação. Na determinação dos níveis de inibidores de proteinases, o tratamento composto por plantas infestadas com ácaros tratados apresentou valor significativamente inferior, em relação aos demais tratamentos. O nível de aldeídos totais foi baixo em todos os tratamentos. Para o T. evansi tratado não houve diferença significativa na taxa de oviposição para os diferentes substratos utilizados. Já para o T. evansi não tratado foi verificada uma maior taxa de oviposição quando utilizado como substrato folhas de plantas pré-infestadas com T. evansi e pré-infestadas com T. evansi com a microbiota reduzida. Tais resultados podem estar relacionados a qualidade nutricional do substrato oferecido. Os resultados advindos deste trabalho vêm contribuir para o conhecimento científico sobre a interação T. evansi-tomateiro, sendo necessária a realização de pesquisas futuras visando à identificação dos microrganismos envolvidos na associação e o estudo de outros parâmetros biológicos.