Proton coupled electron transfer reactions often involve tyrosine residues, because when oxidized, the phenolic side chain deprotonates. Tyrosine Z (YZ) is responsible for extracting electrons in a stepwise fashion from the oxygen evolving-complex in order to build enough potential to oxidize water. This process requires that each step YZ must deprotonate and reprotonate in order to maintain the high midpoint potential that is necessary to oxidize the oxygen-evolving complex, which makes YZ highly involved in proton coupled electron transfer reactions. In this thesis YZ has been studied within oxygen-evolving photosystem II utilizing electron paramagnetic resonance spectroscopy to monitor the tyrosyl radical that is formed upon light excitation. Kinetic analysis of YZ has shed light on the factors that are important for PSII to carry out water oxidation at the oxygen-evolving complex. Most notably the strong hydrogen-bonding network and the midpoint potential of YZ have been shown to be integral aspects of the water splitting reactions of PSII. By studying YZ within oxygen-evolving PSII, conclusions are readily applied to the native system.

American chestnut was once an abundant species that dominated the Eastern U.S. deciduous forests. Although this species is currently functionally extinct due to the chestnut blight, researchers are working on blight-resistant hybrids in hopes of restoring the species. As one potential vector for chestnut reintroduction and dispersal, the reclamation of mine sites are being considered. Recent research has found that reforestation efforts on these reclaimed mine sites provide productive tree growth while also complying with mine-reclamation laws. Understanding how American chestnut performs physiologically on mine sites will aid in the restoration of this species and reclamation of mine sites. The objective of this study was to determine the effect planting treatments have on survival, physiology, and performance of American chestnut. The response of American chestnuts under planting treatments varying in planting method, slow-release fertilizer, a hydrophilic root polymer (Terra-Sorb), and the addition of native forest soil were examined at three sites: a mine site, a quarry, and a greenhouse. Results from this study suggest that fertilizer, hydrophilic root polymers and soil microorganisms produce varying effects on dissimilar sites. Greatest overall survival was found in greenhouse-grown bare-root seedlings. The introduction of fertilizer to the soil substrate lowered emergence and survival of directly-seeded trees at the mine site and quarry, but increased tree growth at all sites and photosynthetic rate at the quarry. However, use of fertilizer resulted in a more negative water potential at the mine site and higher transpiration rate, potentially increasing moisture stress and demand. Increased seed survival, growth, and water status can be accomplished through the use of Terra-Sorb, but only in direct seeded trees on sites with potentially deficient plant-available water. Native forest soil can increase survival in both direct seeded and bare-root planted trees. However, more research is needed on the benefits of native forest soil, as growth and physiological results conflict. It is recommended that site characterization be performed prior to selection of planting treatments. Results of this study can prove beneficial to reclamation specialists with an interest in using mine site reclamation in conjunction with American chestnut restoration.

Magistrantūros studijų darbe pateikiami egzogeninio prolino poveikio žaliosios šerytės augimui ir vystymuisi tyrimų rezultatai. Darbo objektas – žalioji šerytė (Setaria viridis (L.) P. Beauv.). Darbo metodai: IV organogenezės tarpsnyje žaliosios šerytės augalai nupurkšti prolino tirpalais ir auginti programuojamoje auginimo kameroje esant 20/18 °C (diena/naktis) temperatūrai, 16/8 val. (diena/naktis) fotoperiodui, 50 µmol m-2 s-1 apšviestumui. Fotosintezės pigmentai, augalo žalia masė ir sausos medžiagos kiekis nustatyti praėjus 1, 5, 10 ir 15 dienų po purškimo. Esant 35-40% substrato drėgniui augalai nupurkšti 10 mM L-prolino tirpalu ir toliau auginti programuojamoje auginimo kameroje sausros sąlygomis. Fotosintezės pigmentai, augalo žalia masė ir sausos medžiagos kiekis nustatyti po 3 ir 6 sausros dienų. Darbo rezultatai. Žaliosios šerytės augalai sukaupė statistiškai patikimai didesnį chlorofilo a ir karotenoidų kiekį visų tirtų L-prolino koncentracijų poveikyje. Augalus paauginus 10 ir 15 dienų po purškimo nustatytas neigiamas egzogeninio L-prolino poveikis chlorofilo b kiekiui žaliosios šerytės augaluose. Neigiamas egzogeninio L-prolino poveikis žaliai masei nustatytas praėjus 5 ir 10 dienų po purškimo, o paauginus augalus 15 dienų neigiamas L-prolino poveikis augalo žaliai masei beveik eliminavosi. Visos tirtos L-prolino koncentracijos praėjus 15 dienų po purškimo didino sausos medžiagos kiekį žaliosios šerytės augaluose ir spartino generatyvinių organų susidarymą... [toliau žr. visą tekstą] / The objective of this study was to investigate the effect of exogenous proline on growth and development of green millet plants. Object of the work – green millet (Setaria viridis (L.) P. Beauv.). Methods of the work: green millet plants were to spray with proline and growing in growth chamber under 20/18 °C (day/night) temperature, 16/8 h (day/night) photoperiod, 50 µmol m-2 s-1 light intensity. Amount of photosynthetic pigments, fresh mass of plants, dry weight was evaluated after 1, 5, 10 and 15 days after treatment. When substrate humidity was 35-40%, plants were to spray with 10 mM proline and growing in growth chamber under moisture deficit. The results of work. Green millet plants accumulated significantly higher chlorophyll a and carotenoids contents under influence of all proline concentration tested. Negative effect of exogenous L-proline on amounts of chlorophyll b in green millet plants has been obtained after 10 and 15 growing days. Negative effect of exogenous L-proline on fresh mass of green millet plants has been obtained after 5 and 10 growing days, however this effect was almost eliminated after 15 growing days. All tested L-proline concentrations increased dry weight after 15 growing days and accelerated formation of generative organs. After 3 and 6 drought stress treatment, exogenous proline didn’t have any effect on chlorophyll b content, but resulted in increasing of chlorophyll a content. Exogenous proline didn’t have significant effect on dry weight... [to full text]

La recherche de sources d’énergie fiables ayant un faible coût environnemental est en plein essor. L’hydrogène, étant un transporteur d’énergie propre et simple, pourrait servir comme moyen de transport de l’énergie de l’avenir. Une solution idéale pour les besoins énergétiques implique une production renouvelable de l’hydrogène. Parmi les possibilités pour un tel processus, la production biologique de l’hydrogène, aussi appelée biohydrogène, est une excellente alternative. L’hydrogène est le produit de plusieurs voies métaboliques bactériennes mais le rendement de la conversion de substrat en hydrogène est généralement faible, empêchant ainsi le développement d’un processus pratique de production d’hydrogène. Par exemple, lorsque l’hydrogène est produit par la nitrogénase sous des conditions de photofermentation, chaque molécule d’hydrogène constituée requiert 4 ATP, ce qui rend le processus inefficace. Les bactéries photosynthétiques non sulfureuses ont la capacité de croître sous différentes conditions. Selon des études génomiques, Rhodospirillum rubrum et Rhodopseudomonas palustris possèdent une hydrogénase FeFe qui leur permettrait de produire de l’hydrogène par fermentation anaérobie de manière très efficace. Il existe cependant très peu d’information sur la régulation de la synthèse de cette hydrogénase ainsi que sur les voies de fermentation dont elle fait partie. Une surexpression de cette enzyme permettrait potentiellement d’améliorer le rendement de production d’hydrogène. Cette étude vise à en apprendre davantage sur cette enzyme en tentant la surexpression de cette dernière dans les conditions favorisant la production d’hydrogène. L’utilisation de résidus organiques comme substrat pour la production d’hydrogène sera aussi étudiée. / The search for alternative energy sources with low environmental impact is in great expansion. Hydrogen, an elegant and simple energy transporter, could serve as means of transporting energy in the future. An ideal solution to the increasing energy needs would imply a renewable production of hydrogen. Out of all the existing possibilities for such a process, the biological production of hydrogen, also called biohydrogen, is an excellent alternative. Hydrogen is the end result or co-product of many pathways in bacterial metabolism. However, such pathways usually show low yields of substrate to hydrogen conversion, which prevents the development of efficient production processes. For example, when hydrogen is produced via nitrogenase under photofermentation conditions, each hydrogen molecule produced requires 4 molecules of ATP, rendering the process very energetically inefficient. Purple non-sulfur bacteria are highly adaptive organisms that can grow under various conditions. According to recent genomic analyses, Rhodospirillum rubrum and Rhodopseudomonas palustris possess, within their genome, an FeFe hydrogenase that would allow them to produce hydrogen via dark fermentation quite efficiently. Unfortunately, very little information is known on the regulation of the synthesis of this enzyme or the various pathways that require it. An overexpression of this hydrogenase could potentially increase the yields of substrate to hydrogen conversion. This study aims to increase our knowledge about this FeFe hydrogenase by overexpressing it in conditions that facilitate the production of hydrogen. The use of organic waste as substrate for hydrogen production will also be studied.

Physical and functional properties of advanced nano-composite materials and biological structures are determined by self-organized atoms and molecules into nanostructures and in turn by microscopic organization of the nanostructures into assemblies of higher structural complexity. Therefore, microscopes are indispensable tools for structural investigations at various levels of organization. In this work, novel nonlinear optical microscopy methods were developed to non-invasively study structural organization at the nanoscopic and microscopic levels. Atomic organization of semiconductor nanowires, molecular organization of amylose biocrystallites in starch granules, and microscopic organization of several photosynthetic organisms was elucidated. The structure of ZnSe nanowires, key components in many modern nanodevices, was investigated using polarization harmonic generation microscopy. Based on nonlinear optical properties of the different crystal lattices, zinc blende and wurtzite nanowires were differentiated, and the three-dimensional orientation of the zinc blende nanowires could be found. The structure of starch granules, a model biocrystal, important in food as well as health sciences, was also investigated using polarization harmonic microscopy. The study was combined with ab initio calculations using the crystal structures of amylose A and B, revealing that second harmonic signals originate from the hydroxide and hydrogen bonds in the starch granules. Visualization of several photosynthetic organisms including the green algae, Chlamydomonas reinhardtii, two species of cyanobacteria, Leptolyngbya sp. and Anabaena sp., aggregates of light-harvesting pigment-protein complexes as well as chloroplasts from green plants were also explored, revealing that future nonlinear microscopy applications could include structural studies of cell walls, the Chlamydomonas eyespot, and photosynthetic membranes. In this study, several nonlinear optical microscopy modalities were developed for quantitative structural investigations of nano and micro-sized architectures. Non-invasive extraction of crystallographic information in microscopic samples will have a number of potential benefits, for example, in clinical applications, allowing observations of disease states inside tissues without the need for biopsy. Industrial nanotechnology will benefit from fast determination of nanostructures with nonlinear microscopy that will improve quality of nanodevices.

Physical and functional properties of advanced nano-composite materials and biological structures are determined by self-organized atoms and molecules into nanostructures and in turn by microscopic organization of the nanostructures into assemblies of higher structural complexity. Therefore, microscopes are indispensable tools for structural investigations at various levels of organization. In this work, novel nonlinear optical microscopy methods were developed to non-invasively study structural organization at the nanoscopic and microscopic levels. Atomic organization of semiconductor nanowires, molecular organization of amylose biocrystallites in starch granules, and microscopic organization of several photosynthetic organisms was elucidated. The structure of ZnSe nanowires, key components in many modern nanodevices, was investigated using polarization harmonic generation microscopy. Based on nonlinear optical properties of the different crystal lattices, zinc blende and wurtzite nanowires were differentiated, and the three-dimensional orientation of the zinc blende nanowires could be found. The structure of starch granules, a model biocrystal, important in food as well as health sciences, was also investigated using polarization harmonic microscopy. The study was combined with ab initio calculations using the crystal structures of amylose A and B, revealing that second harmonic signals originate from the hydroxide and hydrogen bonds in the starch granules. Visualization of several photosynthetic organisms including the green algae, Chlamydomonas reinhardtii, two species of cyanobacteria, Leptolyngbya sp. and Anabaena sp., aggregates of light-harvesting pigment-protein complexes as well as chloroplasts from green plants were also explored, revealing that future nonlinear microscopy applications could include structural studies of cell walls, the Chlamydomonas eyespot, and photosynthetic membranes. In this study, several nonlinear optical microscopy modalities were developed for quantitative structural investigations of nano and micro-sized architectures. Non-invasive extraction of crystallographic information in microscopic samples will have a number of potential benefits, for example, in clinical applications, allowing observations of disease states inside tissues without the need for biopsy. Industrial nanotechnology will benefit from fast determination of nanostructures with nonlinear microscopy that will improve quality of nanodevices.

No description available.

Almost all life on Earth depends on the products of photosynthesis - the biochemical process whereby solar energy is stored as chemical-rich compounds. The energy of captured photons is transferred through a network of pigment-protein complexes towards the reaction center. The reaction center is responsible for trans-membrane charge separation, which generates a proton motive force which drives all subsequent biochemical reactions. The ultrafast (femtosecond) nature of the primary processes in photosynthesis is the main reason for its astonishing efficiency. On this timescale, quantum effects start to play a role and can appear in measured spectra as oscillations. It has been hypothesized that these are evidence of wave-like energy transfer. To unveil the fundamental principals of ultrafast excitation energy transfer in both natural and artificial light-harvesting systems, advanced spectroscopy techniques have been utilized. Coherent two- dimensional electronic spectroscopy is a state of the art technique which allows the most complete spectroscopic and temporal information to be extracted from the system under study. This technique has allowed us to identify a new photophysical process where the coherence of the initially excited state is shifted to the ground state upon an energy transfer step. Coherence...

Mercury (Hg) is a global pollutant and potent neurotoxin that bioaccumulates in aquatic and terrestrial food webs as monomethylmercury (MeHg). Anaerobic microbes are largely responsible for MeHg production, which depends on the bioavailability of inorganic Hg substrates to methylators. Hg redox cycling pathways such as Hg reduction play a key role in determining Hg’s availability in the environment. Although abiotic photochemical Hg reduction typically dominates in oxic surface environments, Hg reduction pathways mediated by photosynthetic and anaerobic microbes are thought to play an important role in anoxic habitats where light is limited and MeHg production occurs. Currently, the physiological mechanisms driving phototrophic and anaerobic Hg reduction remain poorly understood. The main objective of my thesis is to provide mechanistic details on novel anaerobic and phototrophic Hg reduction pathways. I used a combination of physiological, biochemical and trace Hg analytical techniques to study Hg reduction pathways in a variety of anaerobic and photosynthetic bacteria. I demonstrated that Hg redox cycling was directly coupled to anoxygenic photosynthesis in aquatic purple non-sulphur bacteria that reduced HgII when cells incurred a redox imbalance. I discovered that terrestrial fermentative bacteria reduced Hg through pathways that relied on the generation of reduced redox cofactors. I also showed that sulphur assimilation controlled Hg reduction in an anoxygenic phototroph isolated from a rice paddy. In addition, I developed methods to explore cryptic anaerobic Hg redox cycling pathways using Hg stable isotope fractionation. At its core, my thesis underscores the intimate relationship between cell redox state and microbial Hg reduction and suggests a wide diversity of microbes can participate in anaerobic Hg redox cycling.

Made available in DSpace on 2016-08-12T19:18:28Z (GMT). No. of bitstreams: 1 CristhyanAGC_TESE.pdf: 2158176 bytes, checksum: f755e84c9ed7d1a46f3c72723b738900 (MD5) Previous issue date: 2015-01-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The survey was conducted in Embrapa tropical agribusiness, in Fortaleza, Ceará, and divided into three separate experiments carried out to evaluate the effects caused by heat stress (Experiments I and II) and by combining temperature and high CO2 concentration (Experiment III) on the growth, physiology, biochemistry and flowering hybrid yellow melon "Goldex" grown in the greenhouse. In all three experiments, the design was completely randomized, the two first were made up of 4 treatments and 8 repetitions and the third was in a 2x2 factorial design with 8 replications. In the experiments I and II, plants were placed in germination chambers with different temperatures (30, 34, 38 and 42 °C), simulating a heat stress. However, in the second experiment, the plants were removed from the chambers at 18 DAT and forwarded to a greenhouse up reaching the flowering phase. In the experiment III, the plants were placed in germination chambers and subjected to treatment 34 °C; 34 °C+900 ppm CO2; 42 °C and 42 °C+900 ppm CO2. In the first experiment, it was found in most of the variables that at least one assessment showed significant differences between treatments, which is not observed for stem diameter and fructose. Plant height, leaf number (NF), leaf area (LA), sucrose and fresh and dry shoot and root (MFA, MSA, MFR and MSR) were negatively impacted by temperature 42 °C. For chlorophyll and gas exchange, despite significant differences between treatments of positive or negative trends, were observed variables. Glucose was positively influenced by the temperatures of 38 and 42°C. In the second trial, there was no significant difference between treatments in the variables except the stem diameter. Plant height, NC, AF and fresh and dry shoot and root mass were negatively impacted by a temperature of 42 °C. Chlorophyll and gas exchange showed significant differences between treatments, however, there was no trend, positive or negative, of the variables in the time periods. The flowering was influenced by the treatments, because of the higher temperature, the later the beginning of this phase is observed. In the experiment III, plant height and stem diameter at 7; 14 e 23 DAT significant differences between the temperatures, concentrations of CO2 and interaction entres them, however, the same result was observed on analysis at 0 DAT fact also checked for the trunk diameter at 23 different concentrations in the DAT CO2. NF to 14 e 23 DAT and the AF and fresh and dry shoot were significantly influenced by factors and interaction entres them. Chlorophyll revealed significant differences only between the temperatures at 7 DAT. Already 14 DAT, there were no significant differences of the factors and interaction between them, at 23 DAT significant differences existed in temperatures and interaction of temperature and CO2. Gas exchange were not affected by temperature, and no interaction between factors at 14 DAT, still the same resource, the CO2 enrichment decreased gs and Ci/Cref not influencing the A and E. Have at 21 DAT, the responses were quite different, and the gas and suffered the effects of temperature, CO2 and the interaction between them. While for Ci/Cref and A were not observed significant effects of the factors nor its interaction with the exception of the increase in the in plants under elevated temperature with or without enrichment of CO2, an effect that was also observed in the gas and the leaves and subjected to 42 ºC + 900 ppm de CO2. The Fv/Fm and F0 were influenced only by temperature. Already Fm and qP suffered no effects of the factors nor the interaction between them. The qN showed significant differences for both factors, with no significant effect only the interaction between them. The ETR was affected by concentrations of CO2 and interaction of temperature and CO2. It follows that the melon subjected to high temperatures (38 and 42 °C) up to 24 days after transplanting showed lower growth. This fact was also observed when plants were subjected to initial thermal stress with 42 °C temperature, with a negative effect also on flowering, slowing within 10 days compared to the temperature of 30 °C. The melon showed further growth in plants under the combination of 34 °C+900 ppm CO2 and lower under high temperature, enriched or not CO2. Most physiological analysis of gas exchange and chlorophyll fluorescence were not affected by treatments / A pesquisa foi realizada na Embrapa Agroindústria Tropical, situada em Fortaleza, Ceará, e dividida em três experimentos distintos que tiveram como objetivos avaliar os efeitos causados pelo estresse térmico (Experimentos I e II) e pela combinação temperatura e elevada concentração de CO2 (Experimento III) no crescimento inicial, fisiologia, bioquímica e florescimento do meloeiro amarelo híbrido Goldex cultivados em casa de vegetação. Em todos os três experimentos, o delineamento utilizado foi o inteiramente casualizado; os dois primeiros foram constituídos de 4 tratamentos e 8 repetições e o terceiro foi em esquema fatorial 2x2 com 8 repetições. Nos experimentos I e II, as plantas foram acondicionadas em câmaras de germinação com diferentes temperaturas (30; 34; 38 e 42 ºC), simulando um estresse térmico. Porém, no experimento II as plantas foram retiradas das câmaras aos 18 DAT e encaminhadas para casa de vegetação até tingirem a fase de florescimento. No experimento III, as plantas foram acondicionadas em câmaras de germinação e submetidas aos tratamentos 34 ºC; 34 ºC + 900 ppm de CO2;42 ºC e 42 ºC + 900 ppm de CO2. No experimento I, verificou-se na maioria das variáveis que pelo menos uma avaliação apresentava diferenças significativas entre os tratamentos, fato este não observado para o diâmetro do caule e frutose.Altura da planta, número de folhas (NF), área foliar (AF), sacarose e massa fresca e seca de parte aérea e raiz (MFA, MSA, MFR e MSR) foram influenciados negativamente pela temperatura 42 ºC. A clorofila e as trocas gasosas, apesar das diferenças significativas entre os tratamentos, não foram observadas tendências positiva ou negativa das variáveis. A glicose foi influenciada positivamente pelas temperaturas de 38 e 42 ºC. No experimento II, houve diferença significativa entre os tratamentos nas variáveis, exceto o diâmetro do caule. Altura da planta, NF, AF e massa fresca e seca de parte aérea e raiz foram influenciados negativamente pela temperatura de 42 ºC. A clorofila e as trocas gasosas apresentaram diferenças significativas entre os tratamentos, no entanto, não se observou tendência, positiva ou negativa, das variáveis nos tempos avaliados. O florescimento foi influenciado pelos tratamentos, pois quanto maior a temperatura, mais tardiamente é observado o início desta fase. No experimento III, a altura das plantas e o diâmetro do caule aos 7; 14 e 23 DAT apresentaram diferenças significativas entre as temperaturas, concentrações de CO2 e interação entres eles, no entanto, o mesmo resultado não foi observado na análise aos 0 DAT, fato também verificado para o diâmetro do caule aos 23 DAT nas diferentes concentrações de CO2. O NF aos 14 e 23 DAT e a AF e massas fresca e seca de parte aérea foram influenciadas significativamente pelos fatores e interação entres eles. A clorofila revelou diferenças significativas apenas entre as temperaturasaos 7 DAT. Já aos 14 DAT houve diferenças significativas dos fatores e interação entre eles, aos 23 DAT as diferenças significativas existiram nas temperaturas e interação temperatura e CO2. As trocas gasosas não foram influenciadas pelas temperaturas e nem pela interação entre os fatores aos 14 DAT, ainda na mesma avaliação, o enriquecimento de CO2 diminuiu a gs e a razão Ci/Cref não influenciando a A e E. Já aos 21 DAT, as respostas foram bem divergentes, a gs e a E sofreram efeitos da temperatura, do CO2 e da interação entre eles, ao passo que para a razão Ci/Cref e para A, não foram observados efeitos significativos dos fatores e nem de sua interação, com exceção do aumento da A nas plantas submetidas a elevada temperatura com ou sem enriquecimento de CO2, efeito que também foi observado na gs e E das folhas submetidas a 42 ºC + 900 ppm de CO2. A Fv/Fme a F0 foram influenciadas apenas pelas temperaturas. Já a Fm e a qP não sofreram efeitos dos fatores e nem da interação entre eles. A qN apresentou diferenças significativas para ambos os fatores, não apresentando efeito apenas da interação entre eles. A ETR foi influenciada significativamente apenas pelas concentrações de CO2 e interação temperatura e CO2. Conclui-se que o meloeiro submetido a elevadas temperaturas (38 e 42 ºC) até os 24 dias após o transplantio apresentaram menor crescimento, fato que também foi verificado quando as plantas foram submetidas a estresse térmico inicial com temperatura de 42 ºC, com efeito negativo também no florescimento, retardando em até 10 dias na comparação com a temperatura de 30 ºC. O meloeiro apresentou ainda maior crescimento nas plantas submetidas à combinação de 34 ºC + 900 ppm de CO2 e menor sob elevada temperatura, enriquecida ou não de CO2. Já as análises fisiológicas de trocas gasosas e de fluorescência da clorofila, em sua maioria, não foram influenciadas pelos tratamentos