O presente trabalho apresenta a caracterização química de tintas artísticas brasileiras. Diferente de outras matrizes, os materiais de arte fabricados no Brasil não apresentam informações na literatura que auxiliem em sua conservação e restauração. Amostras de tintas acrílicas e a óleo, nacionais e importadas, foram analisadas pelas técnicas espectroscópicas Raman e infravermelho por transformada de Fourier (FTIR), pirólise acoplada a cromatografia a gás e espectrometria de massas (Py-GC/MS) e microscopia eletrônica de varredura acoplada a espectrometria de fluorescência de raios X (SEM-EDS). Resultados de análise indicam que tintas acrílicas brasileiras são compostas por poliestireno (PS), diferentemente das importadas, compostas por polimetilmetacrilato (PMMA). Enquanto as tintas a óleo apresentam composição semelhante (óleo de linhaça), brasileiras e importadas. Duas pinturas do acervo da Pinacoteca do Estado de São Paulo foram selecionadas para um estudo de caso. As obras apresentam a formação de exsudatos em sua superfície, o que demanda a caracterização química como etapa anterior ao desenvolvimento de uma metodologia de restauração. A obra \"Mandala da saudade\", de Sara Goldman-Belz, é composta por tinta spray (alkyd) e a obra \"Três pessoas\", de Marina Saleme, por tinta a óleo. Os resultados de análise apresentam um primeiro passo no estabelecimento de um banco de dados de materiais artísticos fabricados no Brasil para fins de restauração e perícia criminal de obras de arte. / The current work presents the chemical characterization of Brazilian artistic paints. Differently from other matrices, art materials produced in Brazil lack for information in literature that help in their conservation and restoration. Acrylic and oil paint samples, Brazilian and imported, were analyzed by Raman and Fourier transform infrared (FTIR) spectroscopies, pyrolysis coupled with gas chromatography and mass spectrometry (Py-GC/MS) and scanning electron microscopy coupled with x-ray fluorescence spectrometry (SEM-EDS). Results indicate that Brazilian acrylic paints are composed of polystyrene (PS), differently from imported ones, composed of polymethyl methacrylate (PMMA). Whereas, oil paints present similar composition (linseed oil), Brazilian and imported. Two paintings from the collection of Pinacoteca do Estado de São Paulo were selected for a case study. Both works of art present formation of exudates on their surface, which demands chemical characterization as a previous step to the development of a restoration methodology. The painting \"Mandala da saudade\", by Sara Goldman-Belz, is composed of spray paint (alkyd) and the painting \"Três pessoas\", by Marina Saleme, is composed of oil paint. Results presented in this thesis are the first step towards the establishment of a database for artistic materials produced in Brazil for purposes of art restoration and forensic science.

O presente trabalho apresenta a caracterização química de tintas artísticas brasileiras. Diferente de outras matrizes, os materiais de arte fabricados no Brasil não apresentam informações na literatura que auxiliem em sua conservação e restauração. Amostras de tintas acrílicas e a óleo, nacionais e importadas, foram analisadas pelas técnicas espectroscópicas Raman e infravermelho por transformada de Fourier (FTIR), pirólise acoplada a cromatografia a gás e espectrometria de massas (Py-GC/MS) e microscopia eletrônica de varredura acoplada a espectrometria de fluorescência de raios X (SEM-EDS). Resultados de análise indicam que tintas acrílicas brasileiras são compostas por poliestireno (PS), diferentemente das importadas, compostas por polimetilmetacrilato (PMMA). Enquanto as tintas a óleo apresentam composição semelhante (óleo de linhaça), brasileiras e importadas. Duas pinturas do acervo da Pinacoteca do Estado de São Paulo foram selecionadas para um estudo de caso. As obras apresentam a formação de exsudatos em sua superfície, o que demanda a caracterização química como etapa anterior ao desenvolvimento de uma metodologia de restauração. A obra \"Mandala da saudade\", de Sara Goldman-Belz, é composta por tinta spray (alkyd) e a obra \"Três pessoas\", de Marina Saleme, por tinta a óleo. Os resultados de análise apresentam um primeiro passo no estabelecimento de um banco de dados de materiais artísticos fabricados no Brasil para fins de restauração e perícia criminal de obras de arte. / The current work presents the chemical characterization of Brazilian artistic paints. Differently from other matrices, art materials produced in Brazil lack for information in literature that help in their conservation and restoration. Acrylic and oil paint samples, Brazilian and imported, were analyzed by Raman and Fourier transform infrared (FTIR) spectroscopies, pyrolysis coupled with gas chromatography and mass spectrometry (Py-GC/MS) and scanning electron microscopy coupled with x-ray fluorescence spectrometry (SEM-EDS). Results indicate that Brazilian acrylic paints are composed of polystyrene (PS), differently from imported ones, composed of polymethyl methacrylate (PMMA). Whereas, oil paints present similar composition (linseed oil), Brazilian and imported. Two paintings from the collection of Pinacoteca do Estado de São Paulo were selected for a case study. Both works of art present formation of exudates on their surface, which demands chemical characterization as a previous step to the development of a restoration methodology. The painting \"Mandala da saudade\", by Sara Goldman-Belz, is composed of spray paint (alkyd) and the painting \"Três pessoas\", by Marina Saleme, is composed of oil paint. Results presented in this thesis are the first step towards the establishment of a database for artistic materials produced in Brazil for purposes of art restoration and forensic science.

Establishing plant communities is essential for the restoration of contaminated land. As potential cover materials, fly ash and sewage sludge can prevent formation of acid mine drainage from sulfidic mine waste. The aim of the thesis was to i) screen for plants that can be established in, and prevent leakage of metals and nutrients from sludge on top of ash and tailings, and ii) investigate root growth into sealing layers of ash and sludge. Analyses were performed under laboratory, greenhouse and field conditions using selected plant species to examine the release of Cd, Cu, Zn, N, and P from the materials. Plant physiological responses and interactions with fly ash were also investigated. The data show that plants can decrease metal and nutrient leakage from the materials, and lower the elemental levels in the leachate, but with varying efficiencies among plant species. Plants capable of taking up both nitrate and ammonium were more efficient in preventing N leakage compared with those taking up primarily ammonium. Fast growing plants could raise the pH in acidic sludge leachate, but the initial pH decrease and N leakage was not counteracted by plants. Germination in fresh sludge was problematic, but enhanced by aeration of the sludge. In general, the accumulation of metals in plant shoots was low, especially if ash was located below the sludge. Fresh ash was phytotoxic (e.g., high alkalinity, salinity and metal levels) and induced the activity of stress-related enzymes in shoots. In sealing layers of aged and cured ash, roots could grow if the penetration resistance was low, or into the surface of stronger layers if the surface had become pulverized. The roots caused dissolution of calcium-rich minerals, possibly by exudation of saccharides. Addition of sludge to an ash layer increased root growth, likely due to decreased bulk density and pH, and nutrient addition. In conclusion, with selected plant species and a properly constructed cover, metal and nutrient leaching from the materials and root growth into the sealing layer can be restricted.

A citricultura brasileira apresenta números bastante significativos e expressivos, refletindo a grande importância social e econômica que essa atividade agrícola e industrial tem para a economia do país. O Brasil se destaca como o maior produtor de laranja do mundo. As plantas cítricas podem ser produzidas e multiplicadas de várias formas e a enxertia é a forma de propagação vegetativa mais utilizada comercialmente no Brasil. Dentre os porta-enxertos mais empregados, sobressaem-se o citrumeleiro Swingle e a tangerineira Sunki. Apesar das vantagens presentes nesses dois genótipos, a produção deles está sendo limitada por doenças causadas por fungos e estraminipilas. Em citros, as diversas espécies de Phytophthora são responsáveis pelas doenças gomose e podridões, principalmente de radicelas. P. nicotianae é a espécie que mais afeta as plantas cítricas no Brasil. Não obstante a grande importância de P. nicotianae para os porta-enxertos, alguns mecanismos de resistência ou susceptibilidade ainda requerem mais investigações cientificas. O presente trabalho visa a elucidação de alguns mecanismos de resistência dos citros a P. nicotianae, por meio de estudos fisiológicos e bioquímicos dos porta-enxertos, além da ação de meios de cultura e da qualidade da luz no estudo da fisiologia desse estraminipila. Para isso realizou-se a detecção de alguns equivalentes de compostos fenólicos através da técnica HPLC e o comportamento do genótipo resistente frente à possível inibição da síntese de compostos fenólicos através do tratamento das raízes do porta-enxerto Swingle com Pro-Ca; avaliou-se a atração de zoósporos de P. nicotianae por exsudatos radiculares provenientes dos dois porta-enxertos com o uso de uma armadilha adaptada; verificou-se o comportamento do zoósporo, a motilidade e a zoosporogênese frente à ação de vários compostos fenólicos e testou-se a ação de diferentes meios de culturas e da qualidade da luz no crescimento micelial e na esporulação de P. nicotianae. Os resultados revelaram que a tangerineira Sunki possui maiores quantidades de equivalentes em apigenina que o citrumeleiro Swingle nos dois períodos de avaliação, para o HPLC. O porta-enxerto resistente apresentou mais equivalentes de ácido clorogênico que o genótipo susceptível. Quanto ao Pro-Ca, os equivalentes de compostos fenólicos em raízes de plântulas do citrumeleiro Swingle não diferiram daquelas plantas tratadas com água. O Pro-Ca não inibiu e não desativou a síntese de compostos fenólicos. O uso do anticorpo contra a elicitina \"α-plurivorina\" teve resultados intermediários no consumo de água por plântulas de tangerineira Sunki e a atividade zoosporicida dos compostos fenólicos apresentou resultados variáveis no comportamento da motilidade de zoósporos. Os compostos fenólicos escopoletina e tricetinpentametoxi conseguiram paralisar a motilidade e proporcionaram as menores porcentagens de zoosporogênese. O meio de cultura AA permitiu a maior TCMD, seguido de CA e V8-CaCO3-Ágar. Os maiores valores de esporulação foram encontrados nesses dois últimos meios, respectivamente. Todas as faixas de luzes (e a ausência de luz) estudadas no presente trabalho podem ser avaliadas para o crescimento micelial, porém o tratamento claro foi aquele que mais induziu a esporulação de zoósporos de P. nicotianae. / The Brazilian citrus industry has very significant and expressive numbers, reflecting the great social and economic importance of this industrial and agricultural activity has for the economy. Brazil stands out as the world\'s largest orange producer. Citrus plants can be produced and multiplied in various ways and the bud is the form of vegetative propagation more commercially used in Brazil. Among the most commonly used rootstocks, stand up the Swingle citrumelo and Sunki mandarin. Despite the advantages these two genotypes present, their production is being limited by diseases caused by fungi and estraminipilas. In citrus, all species of Phytophthora are responsible for diseases gummosis and fibrous root rot, especially. P. nicotianae is the kind that affects citrus plants in Brazil. Regardless of the great importance of P. nicotianae for rootstocks, some mechanisms of resistance or susceptibility still require further scientific investigation. This study aims to elucidate some mechanisms of resistance of citrus to P. nicotianae, through physiological and biochemical studies of rootstocks, beyond of action culture media and quality of light in the physiological study of this estraminipila. To this was carried out to detect certain equivalent of phenolic compounds by HPLC and genotype resistant behavior against the possible inhibition of the synthesis of phenolic compounds by treatment of the roots of Swingle rootstock with Pro-Ca; evaluated the zoospores attraction P. nicotianae by exudates from the two rootstocks using an adapted trap; it was the zoospore behavior, motility and zoosporogenesis to the action of various phenolic compounds and tested the action of different culture media and quality of light on mycelial growth and sporulation of P. nicotianae. The results for HPLC revealed that Sunki mandarin has larger amounts of apigenin equivalents Swingle citrumelo in that the two periods. The sturdy rootstock had more chlorogenic acid equivalents of the susceptible genotype. As for the Pro-Ca equivalent of phenolic compounds in Swingle seedling roots not differ from those plants treated with water. The Pro-Ca not inhibited and not disabled the synthesis of phenolic compounds. The use of the antibody against elicitin \"α-plurivorin\" had intermediate results in water consumption by Sunki mandarin seedlings and zoosporicide activity of phenolic compounds showed variable results in the behavior of zoospore motility. The scopoletin and tricetinpentametoxi phenolic compounds managed to paralyze the motility and provided the smallest percentages of zoosporogenesis. The culture medium AA allowed greater DMGR, followed by CA and V8-CaCO3-Agar. The highest sporulation values were found in the last two media, respectively. All lights tracks (and the absence of light) studied in this work can be evaluated for mycelial growth, but the treatment light was that more induced sporulation of zoospores of P. nicotianae.

Empresa Brasileira de Pesquisa Agropecuária / The knowledge of the level of association that occurs between diazotrophic bacteria and fungi arbusculares mycorrhizae (AMF) in grassland ecosystems may be important for the understanding of the changes caused by the addition of fertilizers containing phosphorus (P) and nitrogen (N), in the production and botanical composition of natural pastures. The objective of this study was to evaluate. (a) the occurrence of three genera of diazotrophic bacterial in the root system under fertilization with N and P, and determine the contribution of N via BNF, and (b) evaluate the behavior of the association between AMF and native grasses. Four most abundant grasses in natural grasslands of the Southern Campos in Rio Grande do Sul , Axonopus affinis, Paspalum notatum, Andropogon lateralis and Aristida laevis were grown in pots of 5 kg, in a greenhouse, two studies being conducted (A e B ). In A, two treatments were applied: 50 mg kg-1 soil P and 100 mg kg-1 of soil N (NP) and a control, being evaluated, the number of diazotrophic bacteria of the genera Azotobacter, Azospirillum and Herbaspirillum, and the contribution of BNF was determined by the technique of natural 15N abundance. In B, the treatments consisted of applying 50 mg kg-1 soil P (P), application of 50 mg kg-1 soil P and 100 mg kg-1 of soil N (NP), and a control, in both treatments mycorrhizal colonization was determined. For A, A. laevis demonstrate to be more dependent on biological N fixation than the other species. The grass P. notatum compared with other species demonstrated to be more efficient to absorb available soil N. The dry matter accumulation in shoots of the native species was higher with the application of NP. In B the mycorrhizal colonization was similar between the control, P and NP to the roots of A. lateralis and A. laevis, thus presenting a greater dependence on the mycorrhizal association. Differently, in A. affinis and P. notatum, the mycorrhizal colonization was lower when subjected to fertilization with P and NP, thus presenting a lower dependence. It is concluded for A that fertilization with N and P reduces diazotrophic colonization, increasing the production of dry matter and N content of the tissue. A. laevis showed the highest contribution of biological nitrogen fixation, since P. notatum showed higher N accumulation in soil. In relation to B, A. laevis and A. lateralis have a higher dependence on the mycorrhizal than A. affinis and P. notatum. / O conhecimento do nível de associação que ocorre entre bactérias diazotróficas e fungos micorrízicos arbusculares (FMA) nos ecossistemas campestres, pode ser importante para a o entendimento das alterações provocadas pela adição de fertilizantes, contendo fósforo (P) e nitrogênio (N), na produção e na composição botânica das pastagens naturais. O objetivo do presente trabalho foi avaliar (a) a ocorrência de três gêneros de bactérias diazotróficas no sistema radicular sob fertilização com N e P, e determinar a contribuição de N via FBN, e (b) avaliar o comportamento da associação entre FMAs e gramíneas nativas. Foram utilizadas quatro gramíneas de maior abundância nas pastagens naturais dos Campos Sulinos no Rio Grande do Sul, Axonopus affinis, Paspalum notatum, Andropogon lateralis e Aristida laevis, cultivadas em casa de vegetação, sendo conduzidos dois estudos (A e B). Em A, foram aplicados dois tratamentos: 50 mg kg-1 de solo de P + 100 mg kg-1 de N solo (NP) e uma testemunha, sendo avaliados, o número de bactérias diazotróficas dos gêneros Azotobacter, Azospirillum e Herbaspirillum, e a contribuição da FBN através da técnica da abundância natural de 15N. Em B, os tratamentos consistiram na aplicação de 50 mg kg-1 de solo de P (P); aplicação de 50 mg kg-1 de solo de P + 100 mg kg-1 de N solo (NP), e uma testemunha, em ambos foi determinada a colonização micorrízica. A espécie A. laevis demonstrou ser mais dependente da fixação biológica de N que as demais espécies. A espécie P. notatum em comparação as demais espécies, demostrou ser mais hábil em absorver o N disponível no solo. A matéria seca acumulada da parte aérea das espécies nativas foi maior com a aplicação de NP. A colonização micorrízica foi semelhante entre a testemunha, P e NP para as raízes de A. lateralis e A. laevis, apresentando assim uma maior dependência da associação micorrízica. Diferentemente, nas espécies A. affinis e P. notatum, a colonização micorrízica foi menor quando submetidas a adubação com P e NP, apresentando assim uma menor dependência. Conclui-se assim, que a adubação com N e P reduz a colonização de bactérias diazotróficas, aumentando a produção de matéria seca e teor de N no tecido. A. laevis apresentou a maior contribuição da fixação biológica de nitrogênio, já P. notatum apresentou maior acúmulo de N do solo. As espécies A. laevis e A. lateralis apresentam maior dependência da micorrização que as espécies A. affinis e P. notatum.

La phytoremédiation, la technique de bioremédiation qui utilise les plantes, est considérée comme une technologie « verte » et efficace pour décontaminer des sols pollués aux hydrocarbures. Les plantes agissent essentiellement à travers la stimulation des microorganismes de la rhizosphère, où l’exsudation racinaire semble être le moteur majeur de l’activité microbienne qui s’y déroule. Beaucoup de gènes responsables de l’adaptation bactérienne, dans le sol contaminé ou dans la rhizosphère, semblent être portés par les plasmides conjugatifs et transférés entre les bactéries. Une meilleure compréhension de ce phénomène pourrait améliorer le développement de techniques de manipulation du microbiome rhizosphérique afin d’accélérer la bioremédiation. Le but de cette recherche est d’étudier le plasmidome, soit le contenu total en plasmides, de la rhizosphère de saules provenant d’un sol contaminé aux hydrocarbures. Des analyses métagénomiques, de données obtenues à partir d’un séquençage Illumina, ont été utilisées pour étudier les fonctions portées par les plasmides. Nos résultats ont indiqué un fort effet de la contamination aux hydrocarbures sur la composition des plasmides. De plus, les plasmides contenaient des gènes impliqués dans de nombreuses voies métaboliques, telles que la biodégradation d’hydrocarbures, la production d’énergie, la transduction du signal, le chimiotactisme, et les métabolismes des sucres, acides aminés et métabolites secondaires. À ce jour, c’est la première étude de métagénomique comparative documentant la diversité des plasmides dans un contexte de phytoremédiation. Ces résultats fournissent de nouvelles connaissances sur le rôle du transfert latéral de gènes dans l’adaptation bactérienne dans la rhizosphère et dans le sol contaminé aux hydrocarbures. / Phytoremediation, a bioremediation technique that uses plants, is considered to be an effective and affordable “green technology” to clean up hydrocarbon contaminated soils. Plants essentially act indirectly through the stimulation of rhizosphere microorganisms. Root exudation is thought to be one of the predominant drivers of microbial communities in the rhizosphere and is therefore a potential key factor behind enhanced hydrocarbon biodegradation. Many of the genes responsible for bacterial adaptation in contaminated soil and the plant rhizosphere are thought to be carried by conjugative plasmids and transferred among bacteria. A better understanding of these phenomena could thus inform the development of techniques to manipulate the rhizospheric microbiome in ways that improve hydrocarbon bioremediation. This research aims to study the plasmidome (the overall plasmid content) in the rhizosphere of willow growing in hydrocarbon contaminated and non-contaminated soils, as compared with unplanted soil. Metagenomic analyses based on Illumina sequencing were used to highlight functions carried by plasmids. Our results indicate a strong effect of hydrocarbon contamination on plasmid composition. Furthermore, plasmids harbored genes involved in several metabolic pathways, such as hydrocarbon biodegradation, energy production, signal transduction, chemotaxis, metabolisms of carbohydrates, amino acids and secondary metabolites. To date, this is the first comparative soil metagenomics documenting the plasmidome diversity in a phytoremediation system. The results provide new knowledge on the role of lateral gene transfer in the bacterial adaptation in rhizosphere and in hydrocarbon contaminated soil.

In dieser Arbeit wurden zunächst die Kolonisationen von drei verschiedenen Pflanzengattungen durch den GFP-markierten Bacillus amyloliquefaciens FZB42 mittels confocaler Lasermikroskopie und Elektronmikroskopie verfolgt. Hier konnte gezeigt werden, dass FZB42 alle ausgewählten Pflanzen besiedeln konnte. Bei Arabidopsis- und Maiskeimlingen wurden die Wurzelhaare und Verbindungen, an denen laterale Wurzeln entstehen, durch FZB42 bevorzugt besiedelt. Weiterhin wurden bei Arabidopsis die Spitzen der Primärwurzeln, und bei Mais die Wurzelkerben bevorzugt besiedelt. Bei Lemna wurden FZB42 Zellansammlungen entlang der Furchen, die zwischen den Epidermiszellen der Wurzel liegen, sowie den intrazellulären Hohlräumen an der Blattunterfläche gefunden. Anschließend wurden die Transkriptome von FZB42, der mit Maiswurzelexudat angezogen wurde, mittels Microarray analysiert. Insgesamt wurden 302 Gene, die 8,2 % des Transkriptoms ausmachen, signifikant durch das Wurzelexudat beeinflusst, wobei die Mehrzahl (260 Gene) hochreguliert wurde. Die induzierten Gene, dessen Funktion bereits bekannt ist, sind hauptsächlich an dem Nährstoffwechsel, Chemotaxis und Beweglichkeit, sowie an der Produktion von Antibiotika beteiligt. Auch wurden die Trankriptome von sieben FZB42-Muatnten durch Microarray analysiert. Diese hatten jeweils eine Deletionen in fünf Sigmafaktor-Genen (sigB, sigD, sigM, sigV,and sigX) und zwei globalen Transkriptionsregulator-Genen (degU und abrB). Die Expression vieler Genen wird durch diese Genprodukte beeinflusst. Mögliche Mechanismen, wie diese Faktoren die bakterielle Reaktion auf Wurzelexsudaten beeinflüssen, wurden vorgeschlagen. Schließlich wurden Northernblott-Untersuchungen an möglichen sRNA-Kandidaten durchgeführt, dessen Expression signifikant durch Wurzelexudate beeinflusst wurde. Dabei konnten 6 von 20 vermeintlichen sRNA-Kandidaten betätigt werden. Dies weist auf eine noch unbekannte Rolle der sRNAs bei der Pflanzen-Mikroben-Wechselwirkung. / In this work colonization of three different plants genera, maize, Arabidopsis, and Lemna, by GFP-labeled Bacillus amyloliquefaciens FZB42 in a gnotobiotic system was firtly studied using confocal laser scanning microscopy and electron microscopy. It was shown that FZB42 is able to colonize all these three plants with a specific pattern. Root hairs and the junctions where lateral roots occurred were a preferred area of FZB42 on both maize and Arabidopsis seedlings. On Arabidopsis, tips of primary roots were another favored site of FZB42; while, on maize, the concavities in root surfaces were preferred. FZB42 cells were also able to colonize Lemna, preferably accumulating along the grooves between epidermis cells on roots and the concaved intercellular space on fronds. Secondly, microarray experiments were performed concerning the transcriptomic response of FZB42 to maize root exudates. A total of 302 genes representing 8.2% of FZB42 transcriptome were significantly altered in transcription by the presence of root exudates, the majority of them (260) were up-regulated in expression. The induced genes with known function were mainly involved in nutrition utilization, chemotaxis and motility, and antibiotic production. The transcriptome of seven FZB42 mutants, defective in five sigma factor genes (sigB, sigD, sigM, sigV, and sigX) and two global transcriptional regulator genes (degU and abrB), were also investigated through microarray experiments. A vast number of genes were indentified to be controlled by the protein factors respectively. Possible mechanisms were proposed of how these protein factors are involved in the response to root exudates. Finally, by northern blot existence of six out of 20 small RNA (sRNA) candidates was identified, which were significantly altered in expression by root exudates. This suggests that sRNA may play a hitherto unrecognized role in plant-microbe interaction.

Bacillus amyloliquefaciens FZB42 ist ein frei lebendes Bakterium, das Pflanzenwurzeln besiedelt und das Pflanzenwachstum durch viele verschiedene Wirkmechanismen anregt. In dieser Arbeit wurden die molekularen Grundlagen dieser positiven Wirkungen, die dieses „Pflanzenwachstum fördernde Rhizobakterium“ (PGPR) auf seine Wirte ausübt, untersucht. Um den gegenseitigen Austausch von B. amyloliquefaciens und seinen Wirtspflanzen zu entschlüsseln, wurden umfangreiche Proteomstudien durchgeführt. Es wurden Referenzkarten der extrazellulären und zytosolischen Proteinfraktionen erstellt. Die größte Anzahl an ausgeschiedenen Proteinen konnte während der stationären Phase beobachtet werden. Die identifizierten extrazellulären Proteine gehören verschiedenen Funktionsklassen an, wobei die prominentesten Klassen am Kohlenhydrat-Abbau und den Transport von Molekülen durch die Zellwand beteiligt sind. Die zytosolischen Extrakte von Kulturen, die in 1C-Medium bzw. Mineralmedium angezogen wurden, und in der zweidimensionalen Gelelektrophorese (2 DE) aufgetrennt wurden, ergaben 461 und 245 verschiedene Protein-Einträge. Die erstellten Referenz-Karten wurden anschließend verwendet, um Proteine und Prozesse, in an der Interaktion mit Pflanzen beteiligt sind, zu identifizieren. Dafür wurden die Bakterien Wurzelexudaten von Mais (Zea mays L.) ausgesetzt. Die Proteine aus zwei Stämmen, denen die globalen Transkriptionsregulatoren (Degu, AbrB) und vier Sigma-Faktoren (SigB, SigM, SigV, und SigX) fehlen, wurden ebenfalls untersucht, um ihre Beteiligung an den bakteriellen Reaktionen auf die Wurzelausscheidungen zu analysieren. Zusammenfassend ist dies die erste Studie, die umfangreiche Proteomdaten von Gram-positiven PGPR präsentiert, wobei gleichzeitig die Veränderung der Expression von extrazellulären und zytoplasmatischen Proteinen, nach Zugabe von Wurzelexudaten, ausgewertet wurde. / Bacillus amyloliquefaciens strain FZB42 is a free-living bacterium that competitively colonizes plant roots and stimulates plant growth by many different modes of action. The molecular basis of singular beneficial effects that this Plant Growth-Promoting Rhizobacteria (PGPR) exert on their hosts have been studied. To decipher the molecular cross-talk of B. amyloliquefaciens and its’ host plants as a whole system, an extensive proteomic approach was performed. Reference maps of the extracellular and cytosolic protein fractions were established. The highest number of secreted proteins was observed during stationary growth phase. Identified extracellular proteins belong to different functional classes, with the most prominent classes involved in carbohydrate degradation and transportation of molecules across the cell wall. Cytosolic extracts obtained from cultures grown in 1C and minimal media subjected to the 2 Dimensional Electrophoresis (2 DE), revealed 461 and 245 different protein entries, respectively. Created reference maps were subsequently used to identify proteins and processes involved in the interaction with plants, prior to exposure of bacteria to maize (Zea mays L.) root exudates. The proteomics of two strains lacking expression of genes coding for global transcriptional regulators (degU, abrB) and four sigma factors (sigB, sigM, sigV, and sigX) were also inves-tigated, in order to analyse their involvement in bacterial responses to root exudates. In summary, this is the first study presenting comprehensive proteomics of Gram-positive PGPR, evaluating at the same time changes in protein expression caused by addition of root exudates at the extracellular and cytosolic level.

Florações de microalgas potencialmente nocivas têm aumentado em termos globais e causado impactos na aquicultura, no turismo, na saúde humana e em vários aspectos do funcionamento dos ecossistemas. No Brasil, uma atenção cada vez maior tem sido dada ao tema, em virtude do elevado potencial de nossas águas para o cultivo de bivalves marinhos, um dos grupos de invertebrados mais afetados. Deste modo, a preocupação com a contaminação de consumidores humanos tem gerado projetos voltados para a detecção de espécies potencialmente nocivas e de suas toxinas, mas ainda há poucos estudos sobre interações alimentares entre as espécies de fitoplâncton e zooplâncton. No presente trabalho interações entre dinoflagelados potencialmente nocivos e organismos pertencentes ao zooplâncton marinho foram investigadas em dois capítulos. Capítulo 1: O presente estudo tem como objetivo avaliar a influência do dinoflagelado potencialmente nocivo Alexandrium tamiyavanichii na alimentação e reprodução do copépode planctônico marinho Temora turbinata. Taxas de produção de ovos e sucesso de eclosão naupliar foram estimadas para fêmeas alimentadas em misturas de A. tamiyavanichii e do dinoflagelado Prorocentrum minimum, tratado aqui como alimento controle. Capítulo 2: O principal objetivo deste estudo foi analisar os efeitos de exsudatos de dinoflagelados e células intactas na sobrevivência e mobilidade de táxons zooplanctônicos de uma região subtropical (Ubatuba, Brasil). Efeitos letais foram observados em cinco dos seis táxons testados, três dos quais (náuplios de copépodes, tintinídeos e larvas de gastrópodes) quando expostos a exsudatos e dois (rotíferos e larvas de braquiúros) quando expostos a células intactas. Em adição, larvas de gastrópodes demonstraram comprometimento da mobilidade após exposição à exsudatos. Apenas larvas de poliquetas não foram aparentemente afetadas no curso dos experimentos. / Harmful algae blooms have increased globally and caused impacts in aquaculture, tourism, and human health and in various aspects of ecosystem functioning. In Brazil, increasing attention has been given to this issue, because of the high potential of our waters for marine bivalve farming. Therefore, there are risks of contamination of the crops, and consequent poisoning of the consumers. This concern has led research projects to mostly focus on the detection of potentially harmful species and their toxins, with few initiatives to understand feeding interactions among species of phytoplankton and zooplankton. In this research interactions between potentially harmful dinoflagellates and marine zooplankton were investigated in two chapters. Chapter 1 - This study aims to evaluate the influence of the potentially harmful dinoflagellate Alexandrium tamiyavanichii on Temora turbinata feeding and reproduction. Egg production rates and hatching success were estimated for females fed on mixtures of A. tamiyavanichii and P. minimum. Chapter 2: The aim of this study was to analyze the effects of dinoflagellate exudates and intact cells on the survivorship and mobility of zooplankton taxa from a subtropical location (Ubatuba, Brazil). Lethal effects were observed in five out of six taxa investigated, three of which (copepod nauplii, tintinnids and gastropod larvae) when exposed to dinoflagellate exudates and two (rotifers and brachyuran zoeae) when exposed to intact cells. In addition, gastropod larvae displayed mobility impairment during exposure to dinoflagellate exudates. Only polychaete larvae were not apparently affected during the course of the experiments.

Florações de microalgas potencialmente nocivas têm aumentado em termos globais e causado impactos na aquicultura, no turismo, na saúde humana e em vários aspectos do funcionamento dos ecossistemas. No Brasil, uma atenção cada vez maior tem sido dada ao tema, em virtude do elevado potencial de nossas águas para o cultivo de bivalves marinhos, um dos grupos de invertebrados mais afetados. Deste modo, a preocupação com a contaminação de consumidores humanos tem gerado projetos voltados para a detecção de espécies potencialmente nocivas e de suas toxinas, mas ainda há poucos estudos sobre interações alimentares entre as espécies de fitoplâncton e zooplâncton. No presente trabalho interações entre dinoflagelados potencialmente nocivos e organismos pertencentes ao zooplâncton marinho foram investigadas em dois capítulos. Capítulo 1: O presente estudo tem como objetivo avaliar a influência do dinoflagelado potencialmente nocivo Alexandrium tamiyavanichii na alimentação e reprodução do copépode planctônico marinho Temora turbinata. Taxas de produção de ovos e sucesso de eclosão naupliar foram estimadas para fêmeas alimentadas em misturas de A. tamiyavanichii e do dinoflagelado Prorocentrum minimum, tratado aqui como alimento controle. Capítulo 2: O principal objetivo deste estudo foi analisar os efeitos de exsudatos de dinoflagelados e células intactas na sobrevivência e mobilidade de táxons zooplanctônicos de uma região subtropical (Ubatuba, Brasil). Efeitos letais foram observados em cinco dos seis táxons testados, três dos quais (náuplios de copépodes, tintinídeos e larvas de gastrópodes) quando expostos a exsudatos e dois (rotíferos e larvas de braquiúros) quando expostos a células intactas. Em adição, larvas de gastrópodes demonstraram comprometimento da mobilidade após exposição à exsudatos. Apenas larvas de poliquetas não foram aparentemente afetadas no curso dos experimentos. / Harmful algae blooms have increased globally and caused impacts in aquaculture, tourism, and human health and in various aspects of ecosystem functioning. In Brazil, increasing attention has been given to this issue, because of the high potential of our waters for marine bivalve farming. Therefore, there are risks of contamination of the crops, and consequent poisoning of the consumers. This concern has led research projects to mostly focus on the detection of potentially harmful species and their toxins, with few initiatives to understand feeding interactions among species of phytoplankton and zooplankton. In this research interactions between potentially harmful dinoflagellates and marine zooplankton were investigated in two chapters. Chapter 1 - This study aims to evaluate the influence of the potentially harmful dinoflagellate Alexandrium tamiyavanichii on Temora turbinata feeding and reproduction. Egg production rates and hatching success were estimated for females fed on mixtures of A. tamiyavanichii and P. minimum. Chapter 2: The aim of this study was to analyze the effects of dinoflagellate exudates and intact cells on the survivorship and mobility of zooplankton taxa from a subtropical location (Ubatuba, Brazil). Lethal effects were observed in five out of six taxa investigated, three of which (copepod nauplii, tintinnids and gastropod larvae) when exposed to dinoflagellate exudates and two (rotifers and brachyuran zoeae) when exposed to intact cells. In addition, gastropod larvae displayed mobility impairment during exposure to dinoflagellate exudates. Only polychaete larvae were not apparently affected during the course of the experiments.