Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 163-178).

Mémoire (M.Ress.Renouv.)-- Université du Québec à Chicoutimi, 1993. / Bibliogr.: f. 145-153. Document électronique également accessible en format PDF. CaQCU

Mémoire (M.Ress.Renouv.)-- Université du Québec à Chicoutimi, 1993. / Document électronique également accessible en format PDF. CaQCU

Thesis--University of Jyväskylä. / Text in English; abstract and summary also in Finnish. Thesis t.p. inserted. Includes bibliographical references (p. 69-84).

Thesis--University of Jyväskylä. / Text in English; abstract and summary also in Finnish. Thesis t.p. inserted. Includes bibliographical references (p. 69-84).

Thesis (M.S.)--University of Wisconsin--La Crosse, 2002. / Includes bibliographical references.

Orientador: Ladaslav Sodek / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-04T22:11:00Z (GMT). No. of bitstreams: 1 Brandao_AndreaDias_M.pdf: 910878 bytes, checksum: 3dfc539bfdad7f8ba8ddf4be823bb6b0 (MD5) Previous issue date: 2005 / Resumo: A soja [Glycine Max (L.) Merr.] tem sido um dos modelos mais usados entre as plantas cultivadas para estudar os efeitos de tolerância à hipóxia, já que é tida como uma planta com certo grau de tolerância ao estresse de oxigênio e sua importância também tem um caráter econômico bastante relevante. Condições de hipóxia são comuns na natureza, por exemplo em campos alagados onde a difusão de oxigênio para o sistema radicular fica bastante prejudicada. Quando o estresse ocorre em tempo prolongado as conseqüências se tornam irreversíveis, levando as plantas à morte, uma vez que são tantos os prejuízos em função da injúria por carência de O2 que acarretam mudanças estruturais em organelas, por exemplo. Nesses casos, mesmo que a disponibilidade de O2 seja restabelecida, a planta não consegue se recuperar. Sabe-se, no entanto, que a presença do nitrato no meio contribui de forma a diminuir e/ou retardar as conseqüências desse processo, aumentando assim a tolerância das plantas a esta condição. Não é conhecido, entretanto, como o nitrato exerce seu efeito benéfico na tolerância à hipóxia. Alguns trabalhos vêm mostrando a ativação da enzima RN (redutase do nitrato) sob este tipo de estresse, um fenômeno que torna interessante a sua participação na tolerância induzida pelo nitrato. Por essa razão esse estudo teve os objetivos de: 1) otimizar método para extração da enzima RN em plantas de soja; 2) avaliar o papel da enzima RN em condições de hipóxia do sistema radicular e 3) Avaliar a utilização e metabolismo do nitrato em condições de hipóxia do sistema radicular. As plantas [Glycine max (L.) Merr. cv IAC-17] foram cultivadas em sistema hidropônico e após atingirem o estádio de desenvolvimento adequado (V4) foram submetidas à hipóxia (pela interrupção do fornecimento de ar comprimido e adição de uma camada de óleo mineral na solução nutritiva do vaso). O período de acompanhamento dos experimentos foi de 4 dias, tempo necessário para que o sistema radicular atingisse a hipóxia. Após esse período também foi estudada a recuperação das plantas do estresse, após a remoção da camada de óleo e restabelecimento da aeração. Foi observado que, independente do pH inicial, o pH da solução nutritiva das plantas em normóxia ficou estável em torno de 7,7 e o das plantas hipóxicas próximos a 6,6. O consumo de nitrato na solução nutritiva das plantas normóxicas e das hipóxicas foi praticamente o mesmo, também foi detectada a presença de nitrito na solução nutritiva dos tratamentos sob hipóxia. Durante o período de recuperação à normóxia, o nitrito acumulado na solução foi gradativamente utilizado. Quanto às raízes, observou-se que em normóxia ocorreu acúmulo de nitrato, enquanto que em hipóxia ocorreu a utilização do nitrato endógeno. Também foi observado nessas condições (hipóxia) o aumento na concentração de aminoácidos livres totais e proteínas em relação às plantas em normóxia. Na seiva do xilema de plantas sob hipóxia foi observada diminuição no teor de nitrato transportado, variação na composição dos aminoácidos livres presentes (aumentando significativamente a proporção de Ala e Gaba e diminuindo principalmente Asn) e aumento do teor de aminoácidos livres totais, em relação às plantas em normóxia. Nas plantas recuperadas, apenas o teor de proteínas da raiz e a composição de aminoácidos livres da seiva do xilema retornaram aos valores encontrados nas plantas normóxicas. Não foi detectada a presença de nitrito no sistema radicular e/ou seiva do xilema das plantas. A atividade da enzima RN foi bastante modificada durante o período de hipóxia diminuindo a valores bem baixos, assim como não houve sua ativação durante o estresse. Em apenas 1 dia de recuperação (retorno à normóxia), sua atividade atingiu os valores das plantas normóxicas. Chegou-se à conclusão de que embora o grau de utilização do nitrato pela planta seja semelhante em normóxia e hipóxia, o seu metabolismo é bastante alterado na hipóxia. Ao contrário do que foi relatado na literatura, a atividade da RN caiu bruscamente durante a hipóxia sem que houvesse evidência de ativação. No entanto, esta alteração na atividade da enzima pode explicar as diferenças no metabolismo do nitrato entre normóxia e hipóxia / Abstract: Soybean [Glycine Max (L.) Merr.] has been frequently used as a model system among cultivated plants in order to study the effects of tolerance to hypoxia, in view of its certain tolerance to oxygen deficiency and its relevant economic importance. Hypoxic conditions are common in nature, for example in flooded fields where the diffusion of oxygen to the root system suffers serious restrictions. When the stress occurs over a long period the consequences can become irreversible leading to the death of the plant in view of the many prejudicial factors arising from oxygen deficiency. These can result in structural changes in organelles, for example, from which the plant is unable to recover on return to normal oxygen availability. It is known, however, that the presence of nitrate in the surrounding medium can diminish or retard the consequences of oxygen deficiency, thereby increasing the tolerance of plants this stress. It is not known, however, what underlies the beneficial effect of nitrate in plant tolerance of hypoxia. Some investigations have shown that nitrate reductase is activated under oxygen stress, a phenomenon that might be related to the tolerance effect of nitrate. Therefore, this study had the objective of: 1) to optmize method for extraction of the RN enzyme in soybean plants; 2)evaluating the role of nitrate reductase during hypoxic stress of the root system and 3) evaluating the utilization and metabolism of nitrate under these stress conditions. Soybean plants [Glycine max (L.) Merr. cv IAC-17] were cultivated in a hydroponic system and after reaching the V4 stage of development were subjected to hypoxia by suspending aeration and applying a layer of mineral oil to the surface of the nutrient solution. The treatment was carried out for 4 days, time enough for the system to undergo hypoxia. Recovery from stress was also studied following hypoxia where the layer of mineral oil was removed from the surface and aeration of the solution restarted. The results showed that independent of the initial pH, the pH of the nutrient solution of normoxic plants stabilized at around 7.7 while that of the plants under hypoxia stabilized at 6.6. The consumption of nitrate in the nutrient solution of the normoxic and hypoxic treatments was practically the same, and nitrite was also detected in the nutrient solution of the plants under hypoxia. During recovery to normoxia, the accumulated nitrite was gradually used. As for the roots, it was observed that under normoxia, nitrate accumulated, while under hypoxia endogenous nitrate was consumed. Besides, under hypoxia, there was an increase in total free amino acids and proteins compared to the controls. In the xylem sap a decline in nitrate was found under hypoxia together with changes in the amino acid composition (where alanine and Gaba increased markedly while Asn diminished) and increases in the total amino acid levels. On return to normóxia only the levels of protein in the root and amino acid composition of the xylem sap returned to values found in the normoxic plants. No nitrite could be detected in the root tissue or xylem sap. Large changes in nitrate reductase activity were found during hypoxia, being reduced to low values. Nor was there any evidence for the activation of the enzyme during the stress. In less than one day on returning to normoxia activity returned tonormoxic plants levels. It was concluded that despite the similarity in the amounts of nitrate utilized comparing plants under normoxia and hypoxia, it was evident that its metabolism was quite different under these two conditions. Contrary to that reported in the literature, nitrate reductase activity falls drastically during hypoxia without any evidence for activation of the enzyme. Nevertheless, the alteration in activity could explain the differences in nitrate metabolism observed between for normóxia and hypoxia / Mestrado / Mestre em Biologia Vegetal

L’effet de la température et de la composition du milieu en nutriments sur les propriétés des spores (résistance et germination) de B. cereus a été largement étudié contrairement à l'effet de l'anaérobiose. Or, les cellules végétatives de B. cereus peuvent se retrouver dans une grande variété de milieux naturels avec un faible niveau d'oxygène (intestin, sol, lignes de traitement des aliments…) où la sporulation peut avoir lieu. Les spores produites dans ces conditions anaérobies pourraient donc avoir des propriétés particulières. Dans ce travail, un panel de 18 souches de B. cereus appartenant aux groupes phylogénétiques de II à VII a été étudié pour sa capacité à sporuler en anaérobiose dans un milieu de sporulation approprié que nous avons développé (MODS). En anaérobiose, la capacité de sporulation a été plus faible et plus hétérogène qu’en aérobiose. La souche AH187 a produit le niveau de spores le plus important en anaérobiose, elle a donc été choisie pour étudier les propriétés de ces spores. Les spores produites en anaérobiose étaient plus résistantes à la chaleur humide entre 90°C et 100°C, à 1M de NaOH, 1M d'acide nitreux et à la lumière pulsée. Aucune différence dans la résistance à 5 % de peroxyde d'hydrogène ou à 0.25 mM de formaldéhyde, ni aux UV-C, n'a été observée entre les deux conditions. En présence de L- alanine, les spores produites en anaérobiose germaient plus efficacement que celles produites en aérobiose tandis qu’aucune différence dans la germination n’a été observée en présence d'inosine. Aucune différence dans la taille des spores produites dans les deux conditions n’a été observée par microscopie électronique à transmission. Toutefois, les spores obtenues dans des conditions anaérobies avaient un exosporium endommagé ou dans certains cas un exosporium complètement détaché, contrairement aux spores produites dans des conditions aérobies. Afin de comprendre les différences dans la capacité de sporulation de B.cereus entre les 2 conditions, des PCR en temps réel (RT-PCR) ont été utilisées pour étudier l'expression des gènes de l'initiation de la sporulation spo0A, spo0B, spo0F, KinA et kinB. Les cinétiques d'expressions des gènes spo0A, spo0B, spo0F et KinA avaient la même tendance. Ils étaient caractérisés par une expression plus élevée en anaérobiose par rapport à l’aérobiose au début et à la fin de la phase exponentielle de croissance. En outre, l'expression du gène kinB était caractérisée par une augmentation en anaérobiose par rapport à l’aérobiose pour atteindre un pic entre 4 h (milieu de phase exponentielle) et 6 h (début de phase stationnaire) de croissance. Les gènes spo0A, spo0B, spo0F, KinA et kinB sont exprimés de manière différentielle entre l’aérobiose et l’anaérobiose. Ces données pourraient aider à comprendre la différence de capacité de sporulation de B. cereus entre la condition aérobie et anaérobie / The effect of temperature and nutrient composition of the medium on B. cereus spore properties (resistance and germination) has been extensively studied unlike to the effect of anaerobiosis. Nevertheless, B. cereus vegetative cells can be found in a large variety of natural environments with low oxygen level (intestine, soil, food processing line) where sporulation take place. Spores produced in these anaerobic environments could have particular properties. In this work, a panel of B. cereus strains belonging to phylogenetic groups II to VII was studied for their capacity to sporulate in anaerobiosis in an appropriate sporulation medium we developed (MODS). In anaerobiosis, sporulation ability was lower and more heterogeneous than in aerobiosis. The B. cereus AH187 strain produced the highest level of spores in anaerobiosis, it was therefore chosen to study spore properties. Spores produced in anaerobiosis were more resistant to wet heat from 90°C to 100 °C, 1M NaOH, 1M nitrous acid and pulsed light. No difference in resistance to 5 % hydrogen peroxide or 0.25 mM formaldehyde or UV-C was observed between these two conditions. In the presence of L-alanine, spores produced in anaerobiosis germinated more efficiently than spore produced in aerobiosis. No difference in germination was observed with inosine. No difference in the spores size produced in the two conditions was observed by transmission electron microscopy. However, spores obtained under anaerobic conditions had a damaged exosporium, or in some cases a completely detached exosporium, unlike spores produced under aerobic conditions. To understand differences in sporulation ability between both conditions, Real-time reverse transcription-PCR was used to study the expression the expression of sporulation initiation genes spo0A, spo0B, spo0F, kinA and kinB. The kinetics of gene expression spo0A, spo0B, spo0F and kinA had the same trend. They were characterized by a higher expression in anaerobiosis compared to aerobiosis at the beginning and the end of exponential growth phase. Furthermore, kinB gene expression was characterized by an increase in anaerobiosis compared to aerobiosis to achieve a peak between 4 (middle exponential phase) and 6 (early stationary phase) hours of growth. The spo0A, spo0B, spo0F, kinA and kinB genes are differentially expressed between aerobiosis and anaerobiosis. These data may help to understand the difference in B. cereus sporulation capacity between aerobic and anaerobic condition

Ciliates are also of the most extensively studied and diverse groups of unicellular eukaryotes, and yet, their anaerobic representatives have been largely neglected; in part due to culturing difficulties. Although all main ciliate lineages contain anaerobes, their diversity and evolution of anaerobiosis are especially poorly understood and just starting to gain attention. In fact, Ciliophora is an excellent model group to study adaptations to life in anoxia, since it, apart from the aerobic majority, includes free-living and endobiotic obligately anaerobic lineages, facultative anaerobes, microaerophiles, and microaerotolerant species. The diversity of Metopida, the free-living order of obligately anaerobic class Armophorea, has been partially revised and significantly broadened during the past years, including numerous redescriptions using modern methods, as well as the description of novel families Tropidoatractidae and Apometopidae, several genera, and multiple species. Oxygen plays a crucial role in ATP production via oxidative phosphorylation that takes place in the mitochondrion in most known eukaryotes. Nevertheless, anaerobic ciliates, among many other eukaryotes that have adapted to low oxygen concentrations or even its absence, have modified their mitochondria and energetic metabolism to...

Bradyrhizobium japonicum USDA 143 grew chemoorganotrophically when supplied with exogenous nitrous oxide as the terminal electron acceptor, or as the alternate terminal electron acceptor to nitrate under anoxic conditions. Cell growth and dissimilatory N₂O reduction were significantly inhibited by acetylene when either N₂O or N₂O plus nitrate served as terminal electron acceptor(s). Reduction of nitrous oxide accounted for 20% of the energy for cell growth in cultures supplied with nitrate as the terminal electron acceptor. Nitrous oxide was produced stoichiometrically in cultures supplied with nitrate and acetylene and growth was proportionately reduced compared to cultures supplied with an equal amount of nitrate. Exogenous nitrous oxide delayed the reduction of nitrate in cultures supplied with both electron acceptors. The final cell yield and/or growth rate of the cells were reduced when N₂O was ≥ 15% of the culture flask headspace. Direct amperometric monitoring of nitrous oxide respiration indicated a specific activity of 0.082 ± 0.004 µmoles N₂O/min/mg cell-protein. The respiration was inhibited by azide. A Clark-type electrode with a platinum cathode, and the instrumentation for monitoring hydrogen uptake amperometrically were used to monitor the reduction of N₂O during anaerobic respiration. / Master of Science