<p>A method describing the expression and purification of PSI-N together with fusion protein, using affinity chromatography and TEV protease. Although the method proved successful, optimization is still needed due to partial degradation of PSI-N.</p>

Little is known of the physiological and biochemical mechanisms underlying competitive interactions between alien invasive grasses and native taxa, and how these are affected by resource supply. Consequently, this study compared photosystem II (PS II) function, photosynthetic gas and water exchange, enzyme and pigment concentrations, flowering and biomass accumulation in an indigenous geophyte, Tritonia crocata (L.) Ker. Gawl., grown in monoculture and admixed with the alien grass, Lolium multiflorum Lam., at different levels of water and nutrient supply. Diminished stomatal conductances were the primary cause of reduced net C02 assimilation rates, and consequent biomass accumulation in T. crocata admixed with L. multiflorum at all levels of water and nutrient supply with one exception. These corresponded with decreased soil water contents induced presumably by more efficient competition for water by L. multiflorum, whose biomass was inversely correlated with soil water content. Biochemical impairments to photosynthesis were also apparent in T. crocata admixed with L. multiflorum at low levels of water and nutrient supply. These included a decline in the density of working photosystems (reaction center per chlorophyll RC/ABS), which corresponded with a decreased leaf chlorophyll a content and a decreased efficiency of conversion of excitation energy to electron transport (¥0 / l-^o), pointing to a reduction in electron transport capacity beyond QA~, a decline in apparent carboxylation efficiency and Rubisco content. At low nutrient levels but high water supply, non-stomatal induced biochemical impairments to photosynthesis (decreased RC/ABS, chlorophyll a and Rubisco content) were apparent in T. crocata admixed with L. multiflorum. These attributed to a reallocation of fixed carbohydrate reserves to floral production which increased significantly in T. crocata under these conditions only and associated with a corresponding reduction in the mass of its underground storage organ (bulb). The results of this study did not support the hypothesis that under conditions of low water and low nutrient supply invasive annual grasses would have a lesser impact on the growth and physiology of native geophytes than under resource enriched conditions that favor growth of these grasses. Unresolved is whether resource limitation and allelopathic mechanisms functioned simultaneously in the inhibition of the native geophyte by the alien grass. / Thesis (M. Environmental Science (Ecological Remediation and Sustainable Utilisation))--North-West University, Potchefstroom Campus, 2008.

A method describing the expression and purification of PSI-N together with fusion protein, using affinity chromatography and TEV protease. Although the method proved successful, optimization is still needed due to partial degradation of PSI-N.

Microalgae hold a great potential to serve as feedstocks for biodiesel production. Tropical and subtropical zones have more solar energy than temperate areas for microalgal culturing. However, outdoor high irradiance results in high temperature in the culturing medium, which is damaging if not lethal to most known microalgal species. New microalgae with heat tolerance, high growth rates and high lipid contents are desirable to establish this industry. Four new green microalgae were isolated in southern Taiwan, located in the subtropical zone. All four species are members of the genus Desmodesmus in the family Senedesmaceae. Two of the four species survived at 45¢Jfor 24 hours, with 5 to 13% mortality rates caused by the heat. Lipid contents of two species reached over 50% in dry biomass under nitrogen starvation. Oil accumulation in tion. Oil accumulation in tion. Oil accumulation in tion. Oil accumulation in the four species positively correlates with their photosystem efficiencies during stress treatments (R2=0.90).

The solar energy accumulated by photosynthesis over billions of years is the sole source of energy available on Earth. Photosystem II (PSII) uses the sunlight to split water, an energetically unfavorable reaction where electrons and protons are extracted from water and oxygen is released as a by-product. Understanding this process is crucial for the future development of clean, renewable and unlimited energy sources, which can use sunlight to split water and produce hydrogen and electricity. In order to do so we need to understand how this is solved in plants. I have been focusing on the role of two lumenal proteins associated with the thylakoid membrane PsbO and Cah3, in the water oxidation process. Convincing evidences have been presented supporting the hypothesis that bicarbonate acts as a proton acceptor in the water splitting process in PSII and the lumenal carbonic anhydrase, Cah3, supplies bicarbonate required for this function. The PsbO protein, an important constituent of the water-oxidizing complex, however, its function is still unknown. The PsbO protein undergoes a pH dependent conformational change that in turn influences its capacity to bind calcium and manganese, forming a catalytic Mn4Ca cluster in PSII. We propose that light-induced structural dynamics of the PsbO is of functional relevance for the regulation of proton release and for forming a proton sensing - proton transporting pathway. The cluster of conserved glutamic and aspartic acid residues in the PsbO protein acts as buffering antennae providing efficient acceptors of protons derived from substrate water molecules. Both proteins, Cah3 and PsbO have a conserved S-S bridge, required for proper folding and activity; therefore they are potential targets for red-ox regulation in lumen. / Solenergi som omvandlats av fotosyntesen under miljarder av år är basen för nästan all energi på jorden. Fotosystem 2 använder solljuset till att oxidera vatten, ur energisynpunkt en ofördelaktig process, där elektroner och protoner extraheras från vattenmolekyler vilket ger upphov till syrgas som biprodukt. Förståelsen av denna process är viktig för att vi i framtiden skall kunna utveckla rena och förnyelsebara energislag i obegrensad mängd. Genom att efterlikna fotosyntesprocessen skulle vi i framtiden kunna utvecka artificiella system som använder solljuset till att sönderdela vatten för att producera vätgas eller elektrisitet. För att kunna göra det så måste vi kunna förstå hur dessa processer fungerar i växterna. Min forskning har fokuserat på att förstå funktionen hos två av de proteiner, PsbO och Cah3, som deltar i sönderdelningen av vatten. Jag har visat, för första gången, att ett lumen karboanhydras, Cah3, deltar i regleringen av den process där vatten spjälkas. Jag postulerar att Cah3 underlättar bort transporten av protoner från det vattenoxiderande komplexet genom att generera bikarbonat lokalt, som kan fungera som proton transportör. PsbO proteinet genomgår en pH beroende konformationsförändring vilket påverkar dess kapacitet and binda calcium och mangan som i sin tur formar ett katalytiskt Mn4Ca center i fotosystem 2. Jag föreslår att en ljusberoende strukturförändring av Psbo är av funktionell betydelse för regleringen av protonfrigörandet och formar ett proton-avkännande och proton-transporterande system. Ett kluster av konserverande glutamat- och aspartat-aminosyror i PsbO proteinet fungerar som ett buffrande nätverk för protoner som frigörs vid oxidering av vatten. Båda dessa proteiner innerhåller S-S bryggor ock kan därför vara red-ox reglerade i lumen.

Photosynthesis is the unique biological process that converts carbon dioxide into organic compounds, for example sugars, using the energy of sunlight. Thereby solar energy is converted into chemical energy. Nearly all life depends on this reaction, either directly, or indirectly as the ultimate source of their food. Oxygenic photosynthesis occurs in plants, algae and cyanobacteria. This process created the present level of oxygen in the atmosphere, which allowed the formation of higher life, since respiration allows extracting up to 15-times more energy from organic matter than anaerobic fermentation. Oxygenic photosynthesis uses as substrate for the ubiquitous water. The light-induced oxidation of water to molecular oxygen (O2), catalyzed by the Mn4CaO5 cluster associated with the photosystem II (PS II) complex, is thus one of the most important and wide spread chemical processes occurring in the biosphere. Understanding the mechanism of water-oxidation by the Mn4CaO5 cluster is one of today’s great challenges in science. It is believed that one can extract basic principles of catalyst design from the natural system that than can be applied to artificial systems. Such systems can be used in the future for the generation of fuel from sunlight. In this thesis the light-induced production of molecular oxygen and carbon dioxide (CO2) by PSII was observed by membrane-inlet mass spectrometry. By analyzing this observation is shown that CO2 not only is the substrate in photosynthesis for the production of sugars, but that it also regulates the efficiency of the initial steps of the electron transport chain of oxygenic photosynthesis by acting, in form of HCO3-, as acceptor for protons produced during water-splitting. This finding concludes the 50-years old search for the function of CO2/HCO3− in photosynthetic water oxidation. For understanding the mechanism of water oxidation it is crucial to resolve the structures of all oxidation states, including transient once, of the Mn4CaO5 cluster. With this application in mind a new illumination setup was developed and characterized that allowed to bring the Mn4CaO5 cluster of PSII microcrystals into known oxidation states while they flow through a narrow capillary. The optimized illumination conditions were employed at the X-ray free electron laser at the Linac Coherent Light Source (LCLS) to obtain simultaneous x-ray diffraction (XRD) and x-ray emission spectroscopy (XES) at room temperature. This two methods probe the overall protein structure and the electronic structure of the Mn4CaO5 cluster, respectively. Data are presented from both the dark state (S1) and the first illuminated state (S2) of PS II. This approach opens new directions for studying structural changes during the catalytic cycle of the Mn4CaO5 cluster, and for resolving the mechanism of O-O bond formation. In two other projects the mechanism of molecular oxygen formation by artificial water oxidation catalysts containing inexpensive and abundant elements were studied. Oxygen evolution catalyzed by calcium manganese and manganese only oxides was studied in 18O-enriched water. It was concluded that molecular oxygen is formed by entirely different pathways depending on what chemical oxidant was used.  Only strong non-oxygen donating oxidants were found to support ‘true’ water-oxidation. For cobalt oxides a study was designed to understand the mechanistic details of how the O-O bond forms. The data demonstrate that O-O bond formation occurs by direct coupling between two terminal water-derived ligands. Moreover, by detailed theoretical modelling of the data the number of cobalt atoms per catalytic site was derived.

Little is known of the physiological and biochemical mechanisms underlying competitive interactions between alien invasive grasses and native taxa, and how these are affected by resource supply. Consequently, this study compared photosystem II (PS II) function, photosynthetic gas and water exchange, enzyme and pigment concentrations, flowering and biomass accumulation in an indigenous geophyte, Tritonia crocata (L.) Ker. Gawl., grown in monoculture and admixed with the alien grass, Lolium multiflorum Lam., at different levels of water and nutrient supply. Diminished stomatal conductances were the primary cause of reduced net C02 assimilation rates, and consequent biomass accumulation in T. crocata admixed with L. multiflorum at all levels of water and nutrient supply with one exception. These corresponded with decreased soil water contents induced presumably by more efficient competition for water by L. multiflorum, whose biomass was inversely correlated with soil water content. Biochemical impairments to photosynthesis were also apparent in T. crocata admixed with L. multiflorum at low levels of water and nutrient supply. These included a decline in the density of working photosystems (reaction center per chlorophyll RC/ABS), which corresponded with a decreased leaf chlorophyll a content and a decreased efficiency of conversion of excitation energy to electron transport (¥0 / l-^o), pointing to a reduction in electron transport capacity beyond QA~, a decline in apparent carboxylation efficiency and Rubisco content. At low nutrient levels but high water supply, non-stomatal induced biochemical impairments to photosynthesis (decreased RC/ABS, chlorophyll a and Rubisco content) were apparent in T. crocata admixed with L. multiflorum. These attributed to a reallocation of fixed carbohydrate reserves to floral production which increased significantly in T. crocata under these conditions only and associated with a corresponding reduction in the mass of its underground storage organ (bulb). The results of this study did not support the hypothesis that under conditions of low water and low nutrient supply invasive annual grasses would have a lesser impact on the growth and physiology of native geophytes than under resource enriched conditions that favor growth of these grasses. Unresolved is whether resource limitation and allelopathic mechanisms functioned simultaneously in the inhibition of the native geophyte by the alien grass. / Thesis (M. Environmental Science (Ecological Remediation and Sustainable Utilisation))--North-West University, Potchefstroom Campus, 2008.

Stuttgart, Univ., Diss., 2007.

Chemnitz, Techn. Univ., Diss., 2007.

Techn. Universiẗat, Diss., 1999--Berlin.