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Coat protein-RNA interaction in vivo and the biotechnological use of VLPs
Kratochvílová, Kateřina ; Moravec, Tomáš (advisor) ; Hála, Michal (referee)
The Tobacco mosaic virus (TMV) is a simple and frequently used model virus which has been studied already more over than 130 years. Due to the intensive study of this virus the details of its infectious cycle, genomic information and also the structure of the created viral particle as well as the mechanism of its creation are known today. The process of encapsidation (viral particle formation) is sufficiently described in the in vitro conditions. In the in vitro conditions the origin of assembly (OAS) was also described. The OAS was identified in the coding sequence of the gene for the movement protein (MP). The importance of replication centers (replication factories) has also been supposed. The aim of the diploma thesis was to study the specificity of the interaction of RNA and coat protein in the process of the particle assembly taking place directly inside the plants. The experiments were performed to verify the necessity of presence of OAS sequence in process of initiation of viral encapsidation. The effect of the cell compartmentation on this process has also been studied. Based on several viral systems (the Tobacco mosaic virus, the Potato virus X, the Bean yellow dwarf virus and Cowpea mosaic virus) gene constructs were created. These constructs enables to study this idea at the molecular...
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Endocytosis in plants
Svitáková, Lenka ; Krtková, Jana (advisor) ; Hála, Michal (referee)
Endocytosis is a process typical for eukaryotic cells. It is a mechanism that ensures invagination of the plasma membrane which leads to the creation of a membrane vesicle with specifically selected cargo. The vesicle is then internalized by the cell. Endocytosis has a crucial role in plant growth and development not only thanks to the reception of nutrients from the extracellular space but also by having a huge impact on the membrane composition which influences cellular signalling. There are few types of endocytosis known in plants. They differ from each other by the molecular machinery which ensures their process. The most studied type in plants is the clathrin-mediated endocytosis. Clathrin-coated vesicle formation proceeds through five stages. However, there are less details known about it in plant cells than in animal cells. In the first part of my bachelor thesis I describe recent knowledge of this topic and the most important participants in this process. I also mention the mechanisms of regulations that plant cells use to coordinate clathrin-mediated endocytosis. In the second part I summarize recently discovered types of endocytosis in plants that are independent from clathrin. These are endocytosis associated with microdomains and fluid- phase endocytosis. The details of these endocytic...
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Functions of RAB GTPases and SNARE proteins in post-Golgi trafficking pathways in response to abiotic stress in plants
Moulík, Michal ; Hála, Michal (advisor) ; Mašková, Petra (referee)
Plants as sessile organisms are strongly affected by abiotic and biotic stress factors. Thus, they have developed an array of morfological, biochemical and physiological adaptations to reduce the negative effects of these factors. The membrane trafficking, among others, plays very important role in adaptation to abiotic stress. In my bachelor thesis I have focused on two important protein families involved in this trafficking, namely on RAB GTPases and SNARE proteins. In the first part, the phenomenon of stress is characterized and the strategies how plants cope with the effect of stressors are described, especially the production of reactive oxygen species (ROS) and autophagy. Following chapter deals with an introduction to the membrane trafficking in plants. In the rest of the thesis, I characterize RAB GTPases and SNARE proteins and provide contemporary insight in the mechanism of their function. The aim of the key parts of these chapters is to summarize current knowledge of RAB GTPases' and SNARE proteins' functions in post-Golgi trafficking pathways during response to abiotic stressors or secondary oxidative stress. Key words: plants, abiotic stress, membrane trafficking, secretion, endocytosis, vacuole, RAB GTPases, SNARE proteins
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Leaf structural changes induced by tropospheric ozone.
Češpírová, Zdeňka ; Lhotáková, Zuzana (advisor) ; Hála, Michal (referee)
Tropospheric ozone is an important pollutant formed by the photochemical reactions of ultraviolet radiation and nitrogenous. Recently, its concentration in the air is increasing due to the promoting climate change and antropogenic activities, mainly because of combustion of the fossil fuels. The ozone is highly reactive molecule degrading to the reactive oxygen species having negative effects on the leaf physiological functions and structure. Ozone penetrates the leaf through the stomata, thereafter it reacts with the cells and their singular compartments. The products formed due to ozone degradation are able to cause the disintegration of plasma membranes and other oxidative damage, for example accumulation of plastoglobuli inside the chloroplasts, changes in the shape of chloroplasts, mitochondria or peroxisomes. At the macroscopic level the visible lesions, chloroses and necroses are observed. In addition, the faster senescence of leaves is observed under the effect of ozone. The main aim of present thesis is to review knowledge of the ozone-induced changes in the leaf structure beginning on the cell structure level and ending at the whole plant body level and to explain physiological mechanisms leading to these damages. Key words: ozone, oxidative stress, leaf structure, mesophyll, reactive...
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Techno-economical comparison of the fusing and flame method of laser cutting of steel sheets with a solid-state laser
Vojteková, Lucia ; Hála, Michal (referee) ; Mrňa, Libor (advisor)
The aim of diploma thesis is a comparison of flame and fusion laser cutting of thin metal sheets for ACO Industries k. s. company, where the experimental part was also realized. After consultation with company, the thesis was extended by data samples, which were cut by compressed air for comparison with other two gases. The optimization design of cutting parameters was created by Taguchi method. Afterwards the surface roughness of samples was measured. Based on Ra and Rz5 parameters, the samples were classified to classes based on their accuracy according to ISO 9013 standard. According to results the fusion method of laser cutting is more suitable method, especially for its good quality of cutting edge, which is not affected by the oxidic layer. Fusion method uses cutting speed two times higher than the flame method, which leads to decrease of time and financial consumption of the process despite of its higher hourly rate. In case of laser cutting of material by compressed air the quality of cut is insufficient in comparison with fusion or flame cutting method.
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Utilization of trehalose in orchids: evolution of trehalase genes
Šoch, Jan ; Ponert, Jan (advisor) ; Hála, Michal (referee)
All orchid species studied so far have been shown to participate in orchideoid mycorrhizal symbiosis. Morover, this symbiosis is absolutely vital component of their life cycle. Exchange of nutrients occurs between symbionts where the fungi provides the orchid with energy and carbon supply at least in its early developmental stages. This study focuses on the possible role of trehalose in this transfer. In vitro experiments have showed in five species from three different subfamilies of Orchidaceae family that they can utilize trehalose comparably with sucrose and glucose. Thus, the ability of trehalose utilization seems to be conserved among orchids. Trehalase enzyme activity was localized histochemically in orchid mycorrhizas. The activity strongly colocalized with colonized tissue supporting a hypothesis that trehalose transfer occurs in this site and is mediated by trehalase. Using bioinformatic methods, trehalase gene duplications were identified in many taxons of Embryophyta including three orchid species. Interestingly, highest number of trehalase gene copies was identified in genome of orchid Dactylorhiza majalis. Trehalose utilization, high trehalase activity in mycorrhizas and trehalase gene duplications in some orchids together indicate that trehalose transfer in orchid myccorhizas...
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Renewal of selected vehicle in Volšatrans company
Hála, Michal ; Rathouský, Bedřich (advisor) ; Volšička, Miroslav (referee)
This bachelor thesis deals with the current state of Volšatrans transport company fleet, captures and evaluates its overall condition and subsequently recommends a proposal for its replacement. The change is made on the basis of the multi-criteria decision-making method, namely the TOPSIS method, the result of which is the output showing particular proposals for the replacement of the vehicle. This output may be in practice used by Volšatrans as a recommendation for selection of a suitable vehicle variant on the basis of their requirements.
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Effectors of RAB GTPases and their role in plant secretion
Růžičková, Martina ; Hála, Michal (advisor) ; Motyka, Václav (referee) ; Klíma, Petr (referee)
Rab GTPases are small signaling molecules that play an important role in vesicle trafficking in eukaryotic cells. Correct signaling through small GTPases allows orchestration of vesicle transport among cellular organelles and also to the cell wall providing cell wall material for cell growth and elongation. Engagement of Rab GTPases in the regulation of endomembrane trafficking is one of the evolutionary conserved aspects of secretion regulation. The network of Rab GTPases interaction includes also various downstream effectors. One of them is the exocyst complex involved in vesicle docking at the plasma membrane. It is a complex composed of eight different subunits (Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70 and Exo84). Exocyst was discovered as Sec4p Rab GTPase effector in yeast and also data from animal models describe the Sec15 exocyst subunit as the Rab-interacting partner, but data from plants are missing. On the other hand, numerous studies identified exocyst role in tip growth of pollen tube and root hairs, seed coat formation, cell plate and cell wall formation, hypocotyl elongation, and importantly also PIN auxin efflux carriers recycling and polar auxin transport. There are two paralogues of SEC15 in the Arabidopsis genome, SEC15a and SEC15b, the previous one already shown to be...
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Saccharides as an integral part of plant antioxidative system
Štěpánová, Nikola ; Lipavská, Helena (advisor) ; Hála, Michal (referee)
The formation of reactive oxygen species (ROS) in plants is inevitable part of aerobic cellular metabolism, during which ROS are generated as by-products of basal metabolic pathways. ROS participate in many processes in plants. If the ROS are at normal level, they serve as signal molecules. Under stress, however, the ROS level in plants increases. High ROS accumulation can cause severe oxidative damage of cellular components, but simultaneously it activates other signal pathways controlling defence against oxidative stress. Plants protect themselves against increased level of ROS in different ways. Either they try to prevent increased level of ROS, or they try to detoxify them. Plants utilize for detoxification two efficient antioxidative systems - enzymatic and non-enzymatic ones. The main goal of this work is to put saccharides into context of plant non-enzymatic antioxidative system and summarize saccharides-based ROS scavenging mechanisms, which have been invented so far. Research on mechanisms of ROS scavenging is highly important, because oxidative damage through excessive production of ROS is considered to be significant cause of productivity losses in agricultural crops. These losses are likely to increase as a result of increased stress burden connected with expected climatic changes. Key...
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