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.

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.

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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.