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Inducible RNAi against essential genes of nitrogen metabolism as a tool for control of GM plants
Kobercová, Eliška ; Fischer, Lukáš (advisor) ; Tylová, Edita (referee)
Uncontrolled spreading of genetically modified (GM) plants is one of the main concerns about their cultivation. Inducible RNA interference against an essential gene could be a tool for control of GM plants. After spraying with a chemical inducer, the essential gene will be silenced so the treated GM plant will die. For testing this strategy we chose two key enzymes of nitrogen metabolism, glutamate synthase (GOGAT) and glutamine synthetase (GS). GS processes ammonium ions into glutamine, then GOGAT transfers the amide group from glutamine to 2-oxoglutarate to form two glutamates. GS/GOGAT cycle is the main pathway for assimilation of ammonium ions, which could be toxic to plants in a higher concentration. Disruption of ammonium assimilation during photorespiration causes a strong inhibition of photosynthesis. The aim of this work was to describe the effects of silencing GOGAT and GS genes in Arabidopsis thaliana. To induce silencing, RNAi hairpin constructs under a control of constitutive or estradiol-inducible promoter were prepared. In selected independent transformants with the inducible hairpin against GOGAT, chlorosis and reduced growth were observed after the estradiol treatment in in vitro conditions. However, the spraying with estradiol was tricky, at the whole plant level, the induction of...
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Potassium deficiency in plants - signs and responses
Štočková, Hana ; Tylová, Edita (advisor) ; Konrádová, Hana (referee)
Plants receive K+ mainly from the soil through the root system. In soil, K+ occurs primarily in minerals such as mica and potassium feldspar. The availability of K+ for plant uptake depends on the form in which K+ occurs in soil. There are forms directly available, slowly available, and unavailable for plant and the transition of K+ between these pools may occur. In plant, K+ is very mobile and it occurs in high amount in cells. It is the most prominent cytoplasmic cation. It affects high number of metabolic processes, including photosynthesis, osmoregulation, and activation of enzymes. K+ shortage can cause changes in plant morphology, anatomy and metabolism. K+ deficiency can be manifested by leaf deformation, decreased leaf area, necrosis, short internodes, reduced rate of photosynthesis, etc. K+ deficient plant is also more vulnerable to pathogens and its resistance to abiotic stress factors such as drought, low temperatures, or salinity is decreased. Deficiency of K+ reduces the biomass and overall yield in agricultural crops, so K+ fertilizers, both soil and foliar, are needed. The thesis focuses on the manifestations of K+ deficiency in plants and summarizes the recent findings on mechanisms of K+ deficiency perception and signal pathways leading to the response to this deficiency. Key...
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Plasma membrane - cell wall interactions - adhesion and signalling
Králíková, Dagmar ; Tylová, Edita (advisor) ; Krtková, Jana (referee)
Interaction of the cell wall and plasma membrane is essential for proper cell functioning as it affects, for example, cell growth, water relations or pathogen penetration, and provides protection against external factors. This interaction may take the form of signal exchange or component transport and its necessary prerequisite is the contact between the two structures. The contact is ensured not only by turgor pressure but also by their direct connection. Experimental evidences have long been known to show that this connection indeed exists - for example, Hechtian strands that are visible in the plasmolyzed cell. Another example is band plasmolysis in cells of the endodermis and exodermis with the Casparian strips, where the plasma membrane is still attached to the cell wall at the site of the strip. This attachment contributes to the protective feature of the apoplastic barrier and allows maintaining it even during plasmolysis. The anatomic aspects of the phenomenon have long been investigated, but specific proteins providing this interaction at the molecular level have not yet been experimentally confirmed. The aim of the bachelor thesis is to summarize recent findings about the mechanisms ensuring the physical connection (adhesion) between the plasma membrane and the cell wall in the plant...
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Factors affecting uptake and distribution of uranium in plants.
Krejčová, Daniela ; Lhotáková, Zuzana (advisor) ; Tylová, Edita (referee)
Uranium is a radionuclide, which naturally occurs in Earth's soil in rather an insignificant amount. It is not very dangerous in such small concentration; however, this concentration is rising due to anthropogenic activity, therefore an estimation of its increase is at hand. It is necessary to research possibilities of not only effective, but also ecological extermination of this contamination. Phytoremediation could be an appropriate solution, but this method is still in its beginning stages when it comes to uranium contaminations. That's why we need to study influences of uranium on plants and find out to what extent are they suitable for phytoremediation of areas contaminated by uranium. Phytoremediation methods are based on the abilities of plants to receive and accumulate certain pollutants. The factors that influence the amount of uranium received by a plant can be divided into four groups: plant species, uranium's oxidation levels, pH of of the medium or the substrate and ligand, which is attached to the uranium (Mitchell, 2013). Transportation of uranium to the aboveground part of the plant is also a factor here. Plants need to deal with the accepted uranium; aside from the radiological risks resulting from the characteristics of radioisotope, uranium is also a heavy metal. Chemical...
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The role of saccharides in mycorrhizal associations
Znojová, Kateřina ; Ponert, Jan (advisor) ; Tylová, Edita (referee)
The mycorrhizal symbiotic association is a worldwide phenomenon. In most cases, it is a mutually beneficial coexistence of a fungus and a plant in which the plant receives mineral nutrients from the mycorrhizal fungus (mainly nitrogen, phosphorus, and water), while the fungus takes carbohydrates from the plant. In some cases, the transfer of nutrients may occur in the opposite direction, and the plant can even use the mycorrhizal fungus one- directionally. However, there are still gaps in the understanding of the importance and principles of carbohydrate transfer in mycorrhiza. Recent findings summarized in this thesis show that the carbohydrates that occur in mycorrhizal association include mainly sucrose, glucose, fructose, and trehalose. Their transfer takes place either passively, based on concentration gradient, or actively, using different plant and fungal carriers. At the same time, the transfer and the use of carbohydrates vary in different types of mycorrhizal associations. Key words: ectomycorrhiza, arbuscular mycorrhiza, ericoid mycorrhiza, orchid mycorrhiza, saccharides, transfer, transporters
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Ecophysiological relevance of extreme sensitivity of orchids to nitrates
Figura, Tomáš ; Ponert, Jan (advisor) ; Tylová, Edita (referee)
Many orchid species are seriously endangered at present. Reasons for their disappearing from natural habitats remain often unclear. Orchids depend on mycorrhizal symbiosis in nature, however only little is known about this symbiosis. Seeds of some species do not germinate in vitro, making their cultivation for scientific and rescue purposes impossible. We found that seed germination of one of such reluctant species, Pseudorchis albida, is strongly inhibited by nitrates even at extremely low concentrations. As this species prefers oligotrophic mountain meadows, nitrate-induced inhibition probably take place in natural conditions. Surprisingly, we found similar but slightly weaker inhibitory effect also in typical mesophillic species and even in slightly eutrophic ones. The sensitivity to nitrates correlates with trophy level of species canopy. This inhibitory effect of nitrates could be weakened by application of a range of growth regulators, including auxins, cytokinins and gibberellins, and also by mycorrhizal fungi. The action of nitratereductase is essential for this inhibitory effect of nitrates. Experiments with NO donors, scavengers, and NO quantification are pointing right at NO as compound which perhaps mediates nitrate inhibitory effect. Based on these results, the nitrate inhibitory...
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Involvement of AtKT/HAK/KUP high-affinity transporters in plant K+ and Cs+ uptake
Šustr, Marek ; Tylová, Edita (advisor) ; Maršík, Petr (referee)
Potassium is essential macroelement and large amount of potassium is taken up by plants, because it's the major osmotic of plant cell. Due to various potassium availability in different kinds of soil, plants have evolved transport system that can maintain sufficient K+ uptake between several orders of magnitude of potassium concentration. In Arabidopsis thaliana mechanism of K+ acquisition is well understood and it's known that AtHAK5 is mainly involved in K+ uptake in its very low concentrations. AtHAK5 belongs to KT/HAK/KUP family that consist 13 members in A. thaliana. There's known function for some members for example in auxin transport in root tip (TRH1) or in K+ efflux in stomatal closure (KUP6). In this thesis possible involvement of yet unstudied transporters KUP5, KUP7, KUP9 and KUP10 in K+ acquisition and homeostasis in A. thaliana was investigated. In vitro cultivation showed that kup9 mutant has very short lateral roots in K+ deficiency. On the other hand kup5 mutant showed significantly higher dry weight in K+ deficiency than Col-0. Kup9 phenotype was successfully replicated in subsequent cultivations and anatomy of lateral root apical meristems was investigated. Root tips of kup9 were differently organized in K+ deficiency and they showed signs of early termination of meristems. To...
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Variability and mechanisms of exodermis differentiation in plant roots
Blascheová, Zuzana ; Tylová, Edita (advisor) ; Konrádová, Hana (referee)
Environmental conditions affect the formation of apoplastic barriers (endodermis and exodermis) in roots. This was shown on many species in many research papers. The exodermal layer is more variable in response to stress conditions than endodermal layer. Cadmium toxicity, as many other stresses, induces faster development of apoplastic barriers. Most of research papers published so far, however characterized only the response of main root to this type of stress factor. Lateral roots, an important part of the root system absorptive surface, are neglected and there is not much information about their response to cadmium stress. The pattern of apoplastic barriers development was therefore analysed in main and also in lateral roots of various size and position on maternal root axis. We found significant differences in response to cadmium stress among these different root types. Then we summed up the differences between these types of roots. Short lateral roots were generally more responsive to cadmium stress, cadmium affected root branching as well as differentiation of apoplastic barriers in lateral roots. These results help us to better understand the response of complex roots system to environmental conditions. In the second part of this work, the role of CASP genes in exodermal development was...
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