|
|
Optical properties of the leaf in relation to its anatomical traits
Neuwirthová, Eva ; Albrechtová, Jana (advisor) ; Tomášková, Ivana (referee) ; Písek, Jan (referee)
Plant functional traits at the leaf level are commonly used to predict ecosystem responses to environmental factors and describe global climate change processes at the ecosystem level. Plant functional traits include both leaf biophysical traits (e.g., photosynthetic pigment content and water content) and structural traits (e.g., leaf thickness and proportion of photosynthetic and non-photosynthetic tissues). Leaf biophysical and structural traits can be detected either destructively in the laboratory or non-destructively using leaf optical properties. Although estimating chlorophyll content from leaf optical properties is a well-established methodology, the influence of leaf structure and internal anatomy on leaf optical properties has only been thoroughly studied in the last two decades. The papers included in my thesis and my thesis itself are mostly focused on the study of typical European deciduous trees of temperate and hemiboreal forests with leaves having a dorsiventral structure (i.e., the mesophyll is differentiated into palisade and spongy parenchyma). Furthermore, my thesis includes a study on the effect of leaf surface structural traits on optical properties. In this study, two groups of phylogenetically close herbs with comparable internal leaf structure were used (mutants of...
|
|
|
Function of PsbO isoforms
Duchoslav, Miloš ; Fischer, Lukáš (advisor) ; Špunda, Vladimír (referee) ; Sobotka, Roman (referee)
(English version) Oxygenic photosynthesis is crucial for most forms of the life on the Earth. The splitting of water and evolution of oxygen is conducted by photosystem II (PSII), a multi-subunit pigment- protein complex embedded in the thylakoid membrane. PsbO is an indispensable subunit of PSII, bound to its transmembrane subunits from the luminal side. The main function of PsbO is to stabilise and protect Mn4CaO5 cluster where the water splitting occurs. However, it has probably also some auxiliary functions. These additional functions might be different for isoforms of PsbO proteins, as suggested for Arabidopsis thaliana, which expresses two genes encoding protein isoforms PsbO1 and PsbO2. This thesis studies auxiliary functions of PsbO with a focus on functional differences between PsbO isoforms. We found that besides Arabidopsis thaliana, also many other plant species express two psbO genes. Interestingly, the duplication of psbO gene occurred many times independently, generally at the roots of modern angiosperm families. In spite of this, the PsbO isoforms differ at similar sites in the protein structure, suggesting that similar subfunctionalisation of PsbO isoforms occurred parallelly in various lineages. Biochemical characterisation of PsbO from green alga Chlamydomonas reinhardtii and...
|
|
|
Nuclear transport: its regulation and role in plant cells
Bokvaj, Pavel ; Schwarzerová, Kateřina (advisor) ; Fišerová, Jindřiška (referee)
The nucleocytoplasmic transport represents a crucial checkpoint of all signal pathways leading to the gene expression, the cell cycle maintenance, and RNA export processes. It is mediated by nuclear pore complexes (NPC) anchored in double nuclear membrane. The NPC structure and the basic architecture of the transport regulations are evolutionarily highly conserved across eukaryotic kingdoms; however, some significant differences and specifics exist in plants. In this thesis, I describe the contemporary level of our knowledge of the nucleocytoplasmic transport regulation and its actors in animalia generally and in planta in detail.
|
|
|
Mechanisms of epidermal cells polarization in plants.
Vojtíková, Zdeňka ; Žárský, Viktor (advisor) ; Soukup, Aleš (referee)
Plant epidermal cells form contact area of the plant, they protect it from impacts of surrounding environment and they mediate the communication with its neighbourhood. In the epidermis there are evenly distributed several cell types with quite specialized morphology (pavement cells, trichomes and guard cells) due to the polarization mechanisms. The cytoskeleton and signal molecules of ROP GTPase family promote the polarized growth. Thanks to polarized growth the cells reach their shapes. The cytoskeleton responds to the signal by expanding the cell, helps with targeting of the secretion to the sites of active growth and mediates polarized formation of the cell wall. On the upper side of the epidermis the cuticle and layer of epicuticular waxex is secreted. The secretion of cuticular components is baso-apicaly polarized. This work summarizes the mechanisms of the polarization in plant epidermal cells discoverd untill now.
|
|
|
Tissue-specific knockout of starch synthesis in columella cells of Arabidopsis thaliana and gravitropic response
Bogdan, Michal ; Fendrych, Matyáš (advisor) ; Retzer, Katarzyna (referee)
Since the studies of plant gravitropism by Charles Darwin, the identity of specific sensors of gravity in plants has been uncertain. To this date, statoliths - starch granules in the root tips - are considered to play a key role in gravity sensing. The role of statoliths as organelles that mediate the gravity sensing ability of plant roots is based on research that uses plants which have severely impaired ability to synthesize starch in general or have their cells that contain statoliths removed or damaged. This represents methodical imperfections that give rise to alternative explanations, like disturbed auxin flow due to heavy damage to the root tip or unknown involvement of starch from other parts of the plant in gravity perception. Thanks to advances in the field of CRISPR/Cas9 technology, we are now able to produce tissue-specific mutants that might help with clarification of whether starch granules in the root tip are involved in sensing gravity and if so, how significant is this involvement. This diploma thesis aimed to answer these questions by adapting the tissue-specific CRISPR/Cas9 system and using it for the creation of mutants that are starchless specifically in the columella cells. Using this approach, we generated one tissue non-specific mutant line and three tissue-specific mutant...
|
|
|
The role of exocyst subunit AtEXO70E2 in autophagy and secretion
Moulík, Michal ; Sabol, Peter (advisor) ; Janda, Martin (referee)
Exocyst is a protein complex composed of eight subunits, evolutionarily conserved in yeasts, animals, and plants. The main function of exocyst is to mediate the tethering of secretory vesicles to the plasma membrane. However, the involvement of exocyst in some other processes, especially in autophagy, has been recently discovered. Plant exocyst is specific because most of its subunits have multiple paralogs. The most diversified subunit is EXO70, which is encoded by 23 paralogous genes in Arabidopsis thaliana. In this thesis, I dealt with subunit AtEXO70E2 (AT5G61010), which has been localized to double-membrane compartments considerably reminiscent of autophagosomes. These compartments were named EXPOs (for exocyst-positive organelles) and described as a component of unconventional protein secretion pathways. There are also hints that EXO70E2 could play a role in autophagic processes. However, details of this relationship remained unexplored. For my experiments, I used stably transformed lines of A. thaliana and transiently transformed leaves of Nicotiana benthamiana. I performed numerous colocalization experiments, applied various pharmacological treatments to the studied lines, and analyzed a mutant line in the EXO70E2 gene. According to my observations, protein EXO70E2 is expressed especially...
|
|
|
Dynamics of ALBA proteins in Arabidopsis thaliana evaluated by fluorescence microscopy
Popelářová, Anna ; Honys, David (advisor) ; Vosolsobě, Stanislav (referee)
ALBA proteins were discovered in Archaea more than 30 years ago. They were gradually identified to be well conserved in Eucaryotes as well. A functional dimeric form of these proteins with DNA and RNA-binding capability was claimed in both mentioned domains of organisms. However, their roles diversified during evolution and vary in between organisms. In Archaea, ALBAs are involved in the genome organization and RNA-protein interactions. In Eukaryotes, there are presented two different subfamilies of ALBA proteins - Rpp20 and Rpp25 subfamily. A sole protein from each subfamily was identified in some organisms though they were multiplied in plants, respectively. These proteins can interact with each other and participate in ontogenetic development and stress responses. According to several studies, ALBA proteins were found to be involved in DNA stability maintenance or pre-rRNA splicing in the nucleus of Arabidopsis thaliana. However, they have been shown to play a role in the cellular metabolism and stress responses in cytoplasm. Six ALBA proteins were identified in the genome of A. thaliana, three from each subfamily. In this study, all heterodimeric protein- protein interactions were investigated by the bimolecular fluorescence complementation (BiFC) assay which revealed positive results in...
|
|
|
Functional characterization of Alba-family genes in Arabidopsis thaliana
Kočová, Helena ; Honys, David (advisor) ; Fischer, Lukáš (referee)
(anglicky) Alba-family proteins were identified in Archaea and Eucarya and are classified among the oldest and the most conserved nucleic acid-binding proteins. The binding preferences and roles differ among certain evolution clades. In Crenarchaea they represent chromatin-binding proteins, while their role in RNA metabolism is suggested in Euryarchaea and Eukaryotes. ALBA proteins are well characterized in human, where they play a role in the RNAse P/MRP complex and in unicellular parasites, such as Plasmodium and Trypanosoma, where an involvement in the life cycle regulation is confirmed. In plants, their role is not yet well understood. The aim of this thesis is to increase a knowledge about the Alba-family proteins in the model plant Arabidopsis thaliana. Based on a minimal changes to development and reproduction in single mutants and high sequence similarity, a functional redundancy of the proteins was assumed. For better understanding of the ALBA proteins function, three smaller members of the family were edited by the same metod. The obtained triple mutant showed delay in flowering. ALBA dimer formation was confirmed in many organisms. BiFC method was used to determine Arabidopsis ALBA homodimerization. The data analysis showed potential homodimerization in most of them.
|
|
|
Function study of EXO70H7 and EXO70H8 genes in Arabidopsis thaliana development.
Modráčková, Jana ; Kubátová, Zdeňka (advisor) ; Soukup, Aleš (referee)
Complex Exocyst consists of eight proteins and it is known as a Sec6/8. Its composition is evolutionarily highly conserved amongst all the species. This complex is involved in vesicle trafficking as a part of attaching mechanism to a specific place on the plasma membrane. EXO70 subunit has been found in 23 copies in Arabidopsis thaliana genome. In this study we have been examine paraloges EXO70H7 and EXO70H8. There have been suggestions that these genes are important in development of roots according to the previous studies. We have not been able to identify any significant phenotype within the mutant plants in these genes. There has been studied other mutant appearance during the stress experiments. Most of these experiments did not identify any divergence. Only experiments with germination during stress conditions revealed significantly worse germination of exo70H7 mutant seeds on the medium containing sorbitol. This suggests that mutant seeds have a worse protection against osmotic stress. Significantly worse germination of exo70H8 seeds on the medium with excess NaCl indicates that these seeds incriminate to higher absorption of sodium ions. Analysis of the cell localization of GFP constructs brought knowledge of appearance EXO70H7 and EXO70H8 proteins. EXO70H7 locates in the cytoplasm and in...
|
|
|
Transport and Metabolism of Radio-Labelled Cytokinins in Plant Cells and Tissues
Nedvěd, Daniel ; Ryšlavá, Helena (advisor) ; Vaňková, Radomíra (referee)
Cytokinins are a large group of phytohormones. Since their discovery in the 1950s, they have shown to play a pivotal role in plant physiology. Most studies so far focused on cytokinin action mechanisms and their metabolic regulation. Identification of AtABCG14 and AtPUP14 as cytokinin-specific membrane carriers brought researchers' attention to cytokinin membrane transport, too. In this thesis, we performed experiments with radio-labelled cytokinin tracers. We show that trans-zeatin and isopentenyladenine, two major biologically active cytokinins, are readily transported across the plasma membrane in tobacco BY-2 cell suspension. Making use of mathematical modelling, we show that BY-2 cells possess a membrane transport system with an affinity toward cytokinins. Next, we show that atabcg14 and atpup14 mutations affect cytokinin metabolism in Arabidopsis thaliana plants. Keywords: cytokinin, Arabidopsis thaliana, tobacco BY-2 cell lines, membrane transport, purine permease, ATP-binding cassette, radio-labelling
|
guest ::
