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Interakce teploty a hormonů v růstu Arabidopsis thaliana
Andrýsková, Kristýna
Abstract Due to global warming, plants are exposed to higher temperatures more often and for longer time periods. Exceeding the temperature optimum causes heat stress and affects plant growth and morphology. The most common plant reactions to high temperature are elongation of the hypocotyl, leaf hyponasty, or elongation of leaf petioles. All these morphological changes are collectively referred to as thermo-morphogenesis. The key modulator of this process is the transcription factor phy-tochrome-interacting factor PIF4. It is the main regulator of thermomorphogenesis through which the YUCCA8 gene is activated. YUCCA8 is a biosynthetic enzyme in auxin metabolism that induces auxin accumulation and activates cell elongation. The experimental part was focused on the influence of cytokinin hormones on the growth of the hypocotyl in Arabidopsis thaliana at standard (20 °C) and elevated (29 °C) temperatures. The increased temperature had a positive effect on the growth of the hypocotyl of control plants as well as transgenic lines with ectopic expression of the PIF4 gene. The effect of the application of 10µM trans-zeatin was temperature dependent. At standard temperature, cytokinins had a positive effect on the growth of the hypocotyl of control plants but did not have a significant effect in transgenic lines. However, transgenic lines had elongated hypocotyls compared to wild-type plants at standard temperature. Conversely, at a higher temperature, cytokinins only had an inhibitory effect. Automatic analysis of the abundance of the PIF4 pro-tein using GFP showed that its level within the hypocotyl is uneven with increasing abundance in the direction from the root to the cotyledons. Confocal microscopy confirmed that the inhibitory effect of cytokinins on hypocotyl elongation at higher temperatures is not due to a decrease in the PIF4 protein level.
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Vliv modifikace receptoru ultrafialového světla na jeho funkci
Blechová, Veronika
Ultraviolet radiation has a negative effect on living organisms, but it is also signal modulating growth and development in plants. This thesis is focused on the effects of UV-B light in Arabidopsis thaliana mutants in UV-signalling pathway with modulated cytokinin level. Our data show that UV-B decrease the quantum yield of the PS II (Fv/Fm) in both mutant lines. Decrease of Fv/Fm was accompanied by growth retardation which could be reverted by exogenous application of 1 µM BAP. Determination of the pigments showed that cytokinins interact with UV-B light and this interaction is dependent on the functional UVR8. On the other hand, inhibition of the thermomorphogenesis by cytokinins is independent of the functional UVR8. Experimental work was also focused on the creation of the fusion genes containing modified UVR8 sequence with marker gene eGFP. The correct sequence of the recombinant GFP was confirmed by commercial sequencing.
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Totální syntéza [15N4] cytokininů a jejich využití
BUČEK, Jan
Cytokinins (CKs) and their metabolites and derivatives are essential for cell division, plant growth regulation and development. They are typically found at minute concentrations in plant tissues containing very complicated biological matrices. Therefore, defined standards labelled with stable isotopes are required for precise metabolic profiling and quantification of CKs, as well as in vivo elucidation of CK biosynthesis in various plant species. In this work, eleven [15N]-labelled C6-purine derivatives were prepared, among them five aromatic (4, 5, 6, 7, 8) and three isoprenoid (9, 10, 11) CKs. Compared to current methods, optimized syntheses of 6-amino-9H-[15N5]-purine (adenine, 1) and 6-chloro-9H-[15N4]-purine (6-chloropurine, 3) were performed to achieve more effective, selective and generally easier approaches. The chemical identity and purity of prepared compounds were confirmed by physico-chemical analyses (TLC; HRMS; HPLC-MS; 1H, 13C, 15N NMR). The presented approach is applicable for the synthesis of any other desired [15N4]-labelled C6-substituted purine derivatives.
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Cytokinines and their role in plant cell division, with accent on G2/M transition
Prášilová, Jana ; Ševčíková, Hana (advisor) ; Bíšová, Kateřina (referee)
The eukaryotic cell cycle is well understood mainly in yeasts and animals. Basic regulatory mechanisms, with cyclin-dependent kinases (CDKs) playing crucial roles, are similar in all eukaryotes including plants. CDKs operate mainly at the key cell cycle checkpoints, G1/S and G2/M. Phosphorylation and dephosphorylation of CDKs by kinases and phosphatases have both negative and positive effect. Negative regulator at the G2/M transition is WEE1 kinase which phosphorylates conserved amino acid residues T14 and Y15 of CDK. Phosphatase CDC25 removes this inhibitory phosphate in yeasts and animals and forces cells into mitosis. Plant cell cycle exhibits remarkable differences. Importantly, it is controlled by phytohormones, and some key points of regulation remain obscure - a functional plant homologue of yeast CDC25 phosphatase has not been found in plants yet though Y15 inhibitory phosphorylation by WEE1 kinase blocks mitosis entry in plants as well. Thus, the regulatory mechanism of G2/M transition in plant cells is still to be found. Phytohormones play a key role, not only in the plant cell cycle, but in whole plant development. Interplay between the two groups of phytohormones: auxins and cytokinins, is crucial. Especially cytokinins significantly influence the regulation of G2/M checkpoint. It is...
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Cytokinines and their role in plant cell division, with accent on G2/M transition
Prášilová, Jana ; Ševčíková, Hana (advisor) ; Bíšová, Kateřina (referee)
The eukaryotic cell cycle is well understood mainly in yeasts and animals. Basic regulatory mechanisms, with cyclin-dependent kinases (CDKs) playing crucial roles, are similar in all eukaryotes including plants. CDKs operate mainly at the key cell cycle checkpoints, G1/S and G2/M. Phosphorylation and dephosphorylation of CDKs by kinases and phosphatases have both negative and positive effect. Negative regulator at the G2/M transition is WEE1 kinase which phosphorylates conserved amino acid residues T14 and Y15 of CDK. Phosphatase CDC25 removes this inhibitory phosphate in yeasts and animals and forces cells into mitosis. Plant cell cycle exhibits remarkable differences. Importantly, it is controlled by phytohormones, and some key points of regulation remain obscure - a functional plant homologue of yeast CDC25 phosphatase has not been found in plants yet though Y15 inhibitory phosphorylation by WEE1 kinase blocks mitosis entry in plants as well. Thus, the regulatory mechanism of G2/M transition in plant cells is still to be found. Phytohormones play a key role, not only in the plant cell cycle, but in whole plant development. Interplay between the two groups of phytohormones: auxins and cytokinins, is crucial. Especially cytokinins significantly influence the regulation of G2/M checkpoint. It is...
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Plant hormones homeostasis: auxin and cytokinin crosstalk
Doležálková, Lucie ; Zažímalová, Eva (advisor) ; Dračínská, Helena (referee)
5 AbstractAbstractAbstractAbstract It has been since 19th century when the phytohormones and their crosstalk became one of the central themes in plant biology. Due to advances in molecular biology there is number of metabolic pathways linked to certain groups of plant hormones revealed by now, nevertheless much remains to be determined about their cooperation. The aim of this thesis is to evaluate the effect of exogenous cytokinin application on oat leaf segments (Avena sativa) - both specifically towards other phytohormones levels and generally in physiological context (in relation to their senescence). Our results confirmed increased auxin levels due to exogenous application of selected cytokinins and identified biological activity of trans-zeatin-9-glucoside (previously considered as irreversibly deactivated cytokinin type substance) in respective plant model. (In Czech) Key words: phytohormones, auxin, cytokinin, Avena sativa, senescence assay
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Cytokinins and plant senescence
Pokorná, Monika ; Lipavská, Helena (advisor) ; Kutík, Jaromír (referee)
Senescence is a natural process, which is a part of plant developmental programme. However, the onset of senescence can be greatly influenced by external conditions. An important part of plant senescence is a senescence of leaves manifested by chloroplasts breakdown. That is connected to mobilization of nitrogen contained in proteins, which provide their function. However, the correct timing of natural senescence is substantive for the plant. The initiation as well as the process of natural senescence is influenced by several factors. Senescence is in varying degrees controlled by phytohormones, regulation factors and it is also driven by epigeneticall processes. Leaf senescence can be caused by several exogenous (light conditions, temperature, water and mineral availability, pathogen attack) and internal (carbohydrate levels, hormones, age, developmental stage) conditions. As well as many other developmental stages, senescence is greatly regulated by phytohormones. Ethylene, abscisic acid, salicyl acid and methyljasmonate support leaf aging, on contrary cytokinins delay senescence associated processes. Studying of cytokinin with chlorophyll analysis is an important part of every study of senescence. This work summarizes literature information of regulation of senescence with an accent on the role...
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Changes in cytokinin contents and metabolism in spinach in response to zinc excess in soil
Žižková, E. ; Gajdošová, Silvia ; Pavlíková, D. ; Pavlík, Milan ; Száková, J. ; Motyka, Václav
Changes in endogenous cytokinin (CK) contents and metabolism in spinach (Spinacia oleracea L.) responding to excessive amounts of zinc in soil were studied. Application of three zinc rates in soil (250, 500, 750 mg.kg-1) resulted in a progressive (1.7- to 3.3-times) increase of endogenous CKs in spinach leaves. This increase was predominantly due to accumulation of inactive and storage CK forms (N- and O-glucosides) and was accompanied with an enhanced activity of cytokinin oxidase/dehydrogenase (CKX), the key CK degradative enzyme, especially on the highest zinc rate (750 mg.kg-1 soil). On this rate, maximum endogenous CK concentrations in both control and stressed plants were found at the first and the last samplings, i.e. in the course of very young leaf development and just before or during flowering. The CKX activity of stressed plants exceeded that of controls during the whole development and reached the maxima at the last sampling of treated plants.
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Evaluation of the possible role of cytokinin O-glucosylation in response of tobacco plants to drought stress
Nováková, Marie ; Dobrev, Petre ; Motyka, Václav ; Gaudinová, Alena ; Malbeck, Jiří ; Pospíšilová, Jana ; Martin, R. C. ; Mok, D. W. S. ; Mok, M. C. ; Vaňková, Radomíra
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