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Characterization of Photosystem I in the Red Alga Porphyridium purpureum
KOSMAS, Panagiotis
Photosynthesis, a fundamental biochemical process on planet Earth, has passed through several steps during the course of evolution since beginning of time. Red algae represent a primitive stage of evolution of photosynthesis in eukaryotes, hence they are important for our understanding of functioning and evolution of photosynthetic energy conversion in general. In the current work, isolation and characterization of the supramolecular complex consisting of photosystem I with three antenna complexes from the red alga Porphyridium purpureum is presented. The purification consisted of sucrose gradient centrifugation followed by anion exchange and size-exclusion chromatography. The composition of the PSI-antenna supercomplex was confirmed using electrophoresis and reverse-phase chromatography. Function of the isolated supercomplex was further investigated using time-resolved absorption and fluorescence spectroscopy.
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Analysis of primary photosynthetic processes in conifers: A comparison of selected methods and their possible utilisation for the study of genetic variability
Palovská, Markéta ; Holá, Dana (advisor) ; Albrechtová, Jana (referee)
Conifers are important both ecologically and socioeconomically, however, same parts of their biology are not that well researched. This includes genetics and breeding and partly even physiology. Because quantitative genetic analyzes applied in breeding necessitate an analysis of a large number of samples, and conventional methods of analysis are quite time-consuming, certain parameters describing e.g. the activity of photosynthetic electron-transport chain (ETC) are considered for such use. Several methods of the measurement of the activity of photosynthetic ETC exist, but there are some problems with their usage in conifers. I studied this issue from different points of view in three parts of this thesis. 1) I compared the photosynthetic ETC activity in 8 species of conifers using chlorophyll (Chl) fluorescence measurements on intact needles and polarographic measurements in isolated chloroplasts. Each method brought different information. 2) I measured Chl fluorescence parameters, reflectance spectra and pigment content in 536 genetically defined trees of Pinus sylvestris L. Many parameters showed relatively high genetic variability and heritability. I have also determined the suitability of various reflectance indices to estimate pigment and water content of needles. 3) I have optimized the...
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Ultrastrucutre of beech chloroplasts under the elevated CO2 concentration and different irradiation
Vrbová, Anna ; Albrechtová, Jana (advisor) ; Kutík, Jaromír (referee)
Forest stands may act as important carbon storage places - sinks, due to carbon allocation into both the plant biomass in the process of photosynthesis and the soil. Enhancement of CO2 concentration affects a whole range of plant physiological processes and, thus, it is necessary to study its effect on photosynthetic apparatus - leaf anatomical structure and chloroplast ultrastructure. The first aim of the Thesis was to evaluate changes in chloroplast ultrastructure of common beech (Fagus sylvatica L.) under the effects of both elevated CO2 concentration and different irradiance. The second aim was to evaluate if the anatomical parameters obtained from the middle part of the leaf are representative for the whole leaf blade. The trees were grown in glass domes at the Bílý Kříž experimental site in the Beskids Mountains (Czech Republic), owned by the CzechGlobe Institute. Leaves were sampled in 2010 from juvenile trees, which were planted in 2005 being 5-year old and cultivated since then in ambient (AC; 390 micromol/mol) and elevated (EC; 700 micromol/mol) CO2 concentrations. The EC effect was recorded to be an increased proportion of starch grains in the chloroplast median section and decreased proportion of of intergranal thylakoids (IGT) while the ratio of granal to intergranal thylakoids...
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The role of chloroplasts in the stele of epiphytic orchid roots
Ungrová, Anna ; Ponert, Jan (advisor) ; Lhotáková, Zuzana (referee)
The photosynthesis of the aerial roots of epiphytic orchids has been the subject of numerous studies. However, the roots are always evaluated as a homogeneous structure, even though they actually consist of significantly different areas. This work deals for the first time with the possibility of the spatial distribution of photosynthesis between the root layers, specifically the cortex and the stele. A combination of various microscopic techniques, the histochemical characterization of the apoplastic barriers and the immunohistochemical localization of the photosynthetic enzyme phosphoenolpyruvate carboxylase has been used. The results show that well-developed chloroplasts in the stele probably occur in the subfamily Epidendroideae in all epiphytic representatives, while in the subfamily Vanilloideae they occur occasionally. The ultrastructure of chloroplasts from both areas is systematically different, so it is likely that their functions also differ. Apoplastic barriers are prominent in the roots and differentiate early during root development, which can effectively isolate chloroplasts in the stele from the cortex. Chloroplasts also occur in the sclerenchyma cells of the stele, where were identified hitherto unknown pits in cell walls that could provide gas exchange within the stele....
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Ultrastrucutre of beech chloroplasts under the elevated CO2 concentration and different irradiation
Vrbová, Anna ; Albrechtová, Jana (advisor) ; Kutík, Jaromír (referee)
Forest stands may act as important carbon storage places - sinks, due to carbon allocation into both the plant biomass in the process of photosynthesis and the soil. Enhancement of CO2 concentration affects a whole range of plant physiological processes and, thus, it is necessary to study its effect on photosynthetic apparatus - leaf anatomical structure and chloroplast ultrastructure. The first aim of the Thesis was to evaluate changes in chloroplast ultrastructure of common beech (Fagus sylvatica L.) under the effects of both elevated CO2 concentration and different irradiance. The second aim was to evaluate if the anatomical parameters obtained from the middle part of the leaf are representative for the whole leaf blade. The trees were grown in glass domes at the Bílý Kříž experimental site in the Beskids Mountains (Czech Republic), owned by the CzechGlobe Institute. Leaves were sampled in 2010 from juvenile trees, which were planted in 2005 being 5-year old and cultivated since then in ambient (AC; 390 micromol/mol) and elevated (EC; 700 micromol/mol) CO2 concentrations. The EC effect was recorded to be an increased proportion of starch grains in the chloroplast median section and decreased proportion of of intergranal thylakoids (IGT) while the ratio of granal to intergranal thylakoids...
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Photosystem I: from bacteria to green plants
Palovská, Markéta ; Holá, Dana (advisor) ; Matyska Lišková, Petra (referee)
Type I Photosystems (PS) are characterized by a Fe-S cluster that serves as the terminal electron acceptor. They are present in 4 prokaryotic groups and all photosynthetic eukaryotes. This work summarizes the knowledge on the structure and function of PS I subunits. The simplest PS (homodimeric; only 2 subunits) is present in Heliobacteria. PS of green sulphur bacteria is also homodimeric and consists of 4 proteins including a membrane-bound cytochrome. The homodimeric PS of Candidatus Chloracidobacterium thermophilum is functional in aerobic conditions. The reaction center of the well-characterized PS I of Cyanobacteria has a heterodimeric structure; the same applies for the eukaryotic PS I. Cyanobacterial PS consists of 12 proteins and forms trimers under low light conditions. The trimer formation is completely suppressed in green plants. Their PS I (at least 15 proteins) is adapted to bind light-harvesting membrane proteins. The PS I of Glaukophyta and Rhodophyta is slightly different from green plants. Various hypotheses about the evolutionary origin of PS I exist. The last common ancestor of all PSs probably shared features of both Type I and Type II. The purpose and the origin of a PS I heterodimer is also discussed in this work. A further understanding of PS I evolutionary history would be helped by...
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Analysis of primary photosynthetic processes in conifers: A comparison of selected methods and their possible utilisation for the study of genetic variability
Palovská, Markéta ; Holá, Dana (advisor) ; Albrechtová, Jana (referee)
Conifers are important both ecologically and socioeconomically, however, same parts of their biology are not that well researched. This includes genetics and breeding and partly even physiology. Because quantitative genetic analyzes applied in breeding necessitate an analysis of a large number of samples, and conventional methods of analysis are quite time-consuming, certain parameters describing e.g. the activity of photosynthetic electron-transport chain (ETC) are considered for such use. Several methods of the measurement of the activity of photosynthetic ETC exist, but there are some problems with their usage in conifers. I studied this issue from different points of view in three parts of this thesis. 1) I compared the photosynthetic ETC activity in 8 species of conifers using chlorophyll (Chl) fluorescence measurements on intact needles and polarographic measurements in isolated chloroplasts. Each method brought different information. 2) I measured Chl fluorescence parameters, reflectance spectra and pigment content in 536 genetically defined trees of Pinus sylvestris L. Many parameters showed relatively high genetic variability and heritability. I have also determined the suitability of various reflectance indices to estimate pigment and water content of needles. 3) I have optimized the...
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