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Význam rostlinných proteinů z podrodiny ABCB pro transport auxinu
Kubeš, Martin ; Zažímalová, Eva (advisor) ; Hejátko, Jan (referee) ; Reinöhl, Vilém (referee)
Polar auxin transport provides essential directional and positional information for many developmental processes in plants. At the cellular level, it is realized by both passive diffusion and the active transport through the membrane proteins - AUX1/LAXes, PINs and ABCBs. The aim of this thesis was to characterize the role of ABCB1, ABCB4 and ABCB19 proteins in polar auxin transport using transformed tobacco BY-2 cell lines. It was shown that the plasma membrane (PM) localization of the ABCB1, 4 and 19 is not polar. The ABCB4 was also more stable on PM after the treatment with auxin influx inhibitors; making use of ABCB4-cell line helped to uncover new characteristics of markers of endocytosis - the FM- dyes. The induction of ABCB19 has led to a decrease in 3 H-NAA accumulation with characteristic auxin starvation phenotype, similar to PIN7 overexpressing cell line, that could be rescued in case of PIN7 cell line by application of the auxin efflux inhibitor NPA; however, the accumulation of auxin in ABCB19-overexpressing cell line was less sensitive to NPA and the rescue of the auxin starvation phenotype was ineffective. Importantly, unique property of the ABCB4 was demonstrated: It displayed dual, auxin-concentration-dependent auxin transport activity in Arabidopsis roots, tobacco BY-2 and yeast cells. The...
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Characterization of factors participating in regulation of intracellular dynamics of auxin carriers
Lacek, Jozef ; Zažímalová, Eva (advisor) ; Ibl, Verena (referee) ; Nodzynski, Tomasz (referee)
Plants are known to adjust the orientation of their organs, shoot and root, to ensure maximal energy generation and nutrient uptake, but also to avoid toxic growth conditions. Directional growth regulation depends on asymmetric plant organ growth and it is crucial to ensure plant survival. It is orchestrated on cellular level in concert with exogenous and intrinsic signals. Even though tropistic growth responses of plants were described by Darwin on macroscopic level already in 1880, now it is necessary to understand molecular mechanisms that underpin efficient modulation of directional plant growth. During my studies I focused on factors that modulate directional root growth regulation. The root is a complex, three-dimensional object, which continuously modifies its shape and growth path. Since the root needs to expand its surface to supply the plant with nutrients and water, it is important to understand how roots cope with changing growth conditions while exploring the soil. If the root cannot manage to grow through soil efficiently, mechanical impedance and lack of resources will also restrict shoot growth as well. Manifold signaling pathways coordinate the complex processes that underpin efficient root growth, including those modulated by phytohormones, sugars, flavonoids and other...
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Mechanizmus transportu cytokininů přes buněčnou membránu a jejich metabolizmus v buňkách tabákové suspenzní kultury BY-2
Klíma, Petr ; Zažímalová, Eva (advisor) ; Galuszka, Petr (referee) ; Procházka, Stanislav (referee)
MECHANISM OF CYTOKININ TRANSPORT ACROSS PLASMA MEMBRANE AND THEIR METABOLISM IN TOBACCO BY-2 CULTURED CELLS Mgr. Petr Klíma / Abstract of Ph.D. Thesis / Prague 2011 Cytokinins (CKs) are plant hormones that play a major role in a number of developmental processes in plants. Those include promotion of cell division, active growth and differentiation, and maintenance of sink-source relationships, as well as control of environmental stress responses. Native CKs are low-molecular derivatives of adenine which seem to act either as paracrine or as long-distance signals. Due to their numerous physiological effects, plants have to precisely control the occurrence of bioactive CK molecules on the levels of the whole plant, its organs, tissues as well as single cells. To achieve this, a concerted action of metabolism and transport processes is required. Studies of the kinetics of CK translocation across plasma membrane in BY-2 suspension-grown tobacco cells suggested the existence of energy-dependent, partially selective transport routes for CK bases and CK ribosides. HPLC analysis of the metabolites of accumulated CKs pointed at their fast degradation or metabolic conversion into physiologically inactive forms. The prevalent ways of inactivation were the degradation to adenine and phosphorylation or phosphoribosyl...
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Hormonal regulation of plant cell division and elongation
Petrášek, Jan ; Zažímalová, Eva (advisor) ; Griga, Miroslav (referee) ; Procházka, Stanislav (referee)
a I I T T T I I a I I a T I T I t r T T I Jml PnrnÁŠnx Suvnalnv SuuuuARyoF DtssERTATtoNTHEsts This thesisis focusedon thehormonalregulationof plantcelldivisionand growth.Both these processes are crucial in shaping sessile plant body. The manifestationof directionalityin ce||p|ateformationduringcýokinesisas we||as in celle|ongationcou|d be found in a form of various shapes of cells, organs and whole plants.Equal and inequal cell division,cell elongationand isodiametriccell growth- they all allow establishing,formingand maintainingcharacterof tissues,organsand whole bodies duringplantdevelopment. Scientificresearchhas alwaysbeenfocusedon regulatorycompounds,the actionof which induces detectable and observable responses. By uncovering the exact mechanisms of their mode of action, many intermediatemessengers were found. However,classicalconceptof "majorregulators'surviveduntilthesedays.lndeed,plant hormones, small organic molecules with wide range of effects, are typical representativesof such compounds,which are studiedalready around one hundred years.They are synthesisedand utilisedin varioustissuesthroughoutthe wholeplant and theycouldbe transportedin vasculatureto longdistances. Auxins and cýokinins - regu|atorsof plant development The researchof recentyears contributedremarkablyto the understandingof...
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AUXIN BINDING PROTEIN 1 (ABP1) and its role in the auxin management in plant cells
Čovanová, Milada ; Zažímalová, Eva (advisor) ; Lüthen, Hartwig (referee) ; Reinöhl, Vilém (referee)
Conclusions The role of AUXIN BINDING PROTEIN 1 (ABP1) in the auxin management in plant cells was followed using simplified model material of suspension-cultured cells of tobacco BY-2 line. ABP1 is a putative auxin receptor considered to mediate fast non-genomic responses to auxin and it can be involved in every aspect of the regulation of auxin responses, metabolism and transport. There are four major conclusions that could be made based on the results presented in this thesis: 1) Auxin binding protein 1 mediates both cell division and expansion in tobacco BY-2 cells. In standard cultivation conditions or at lower concentrations of 2,4-D in culture medium, ABP1 overexpression had no detectable impact on cell division, cell elongation or cell growth.. 5- times increased 2,4-D concentration stimulated weakly cell elongation. . Antisense suppression of ABP1 expression resulted in disturbance in both cell expansion and cell division intensity, suggesting that ABP1 is essential for the control of balance between cell division and cell elongation during the growth cycle. ABP1 is localized in endoplasmic reticulum of cells cultivated in standard medium supplemented with 1 μM 2,4-D and it appeared also at the plasma membrane following the IAA application. 2) ABP1 mediates intercellular auxin transport. Cells...
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Rodina auxinových přenašečů PIN: funkční a evoluční pohled
Skůpa, Petr ; Zažímalová, Eva (advisor) ; Hejátko, Jan (referee) ; Opatrný, Zdeněk (referee)
Growth and development of plant body is dependent on correct and effective integration of information about current deployment of its body parts, as well as on perception and transduction of inputs from environment. Multiple developmental processes within plant body are determined by specific and tightly controlled distribution of molecule with unique signaling mission within plant development - auxin. Spatial distribution of auxin is co-determined by plethora of tightly controlled processes, and the polar auxin transport plays unique role among them. PIN proteins are the plant-specific family of secondary transporters driving movement of auxin across membranes. With their frequent asymmetrical localization within cells, specific expression patterns in developing tissues and their reactiveness to external cues they secure unique, dynamic and asymmetric distribution of auxin within the plant body. This dissertation thesis is focused on characterization of the role different PIN proteins play in determining cellular auxin homeostasis and consequent formation of auxin gradients. Controlled overexpression of PIN proteins in tobacco cells showed, that PIN4 and to some extent also PIN6, function as the direct auxin efflux carriers. In the cellular auxin transport they play the role analogous to other...
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The role of tubulin transport between the nucleus and the cytoplasm
Bokvaj, Pavel ; Schwarzerová, Kateřina (advisor) ; Zažímalová, Eva (referee)
Tubulin is the basic building block of microtubules which ensure intracellular transport, morphological changes, the connection of proteins of metabolic pathways, the segregation of chromosomes during mitosis and many other essential processes in plant cells. The localization of tubulin regarding nucleus is strictly controlled and during the interphase, tubulin is usually not present in the nucleus. As we previously showed, one of the exceptions of the presence of tubulin in the nucleus is the cold stress condition. Also, we discovered several plant-specific nuclear export sequences (NES) in tubulin molecules: one in α-tubulin and two in β-tubulin. In this work I found that double mutation of both functional β-tubulin NESes cause slight accumulation of the protein in the nuclei. However, the phenotype of the transgenic plants expressing β-tubulin with mutations is probably not affected by the presence of the mutated protein. The effect of the expression of the mutated β-tubulins was observed in Nicotiana tabacum BY-2 cells as well. The expression of β-tubulin carrying a single NES3 mutation or double mutation of both NESes bring changes of the division activity of the cells, while the NES2 mutation does not have any effect. Furthermore, it was discovered that the 0řC cold tr eatment does not cause...
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Molecular mechanism of transport of plant hormone auxin into cells
Laňková, Martina ; Zažímalová, Eva (advisor) ; Stiborová, Marie (referee) ; Opatrný, Zdeněk (referee)
The non-uniform distribution of the plant growth regulator (phytohormone) auxin is known to mediate many fundamental processes in plant development. Auxin is transported through the plant body either via vascular pathways or from cell to cell by specialized polar auxin transport machinery. This machinery consists of a balanced system of passive diffusion combined with the activities of auxin influx and efflux carriers. This work is focused on the processes that are involved in the uptake of auxin into plant cells. On the basis of molecular-biological and biochemical characterization, the function as an auxin influx carrier was confirmed for PaLAX1 protein from wild cherry (Prunus avium). The sequences of isolated cDNA of the PaLAX1 gene and of its protein product are highly similar to both the cDNAs and the corresponding protein products of the AUX1/LAX-type genes, coding for putative auxin influx carriers in model plant A. thaliana. On the level of organs and single cells, we have shown that the overproduction of PaLAX1 in transgenic lines resulted in an increase of the content of native auxin indole-3-acetic acid as well as of the uptake of synthetic auxin, 2,4-dichlorophenoxyacetic acid. Further, the mechanism of action of putative auxin influx inhibitors 1-naphthoxyacetic acid (1-NOA),...
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Hormonal regulation of plant cell division and elongation
Petrášek, Jan ; Zažímalová, Eva (advisor) ; Griga, Miroslav (referee) ; Procházka, Stanislav (referee)
a I I T T T I I a I I a T I T I t r T T I Jml PnrnÁŠnx Suvnalnv SuuuuARyoF DtssERTATtoNTHEsts This thesisis focusedon thehormonalregulationof plantcelldivisionand growth.Both these processes are crucial in shaping sessile plant body. The manifestationof directionalityin ce||p|ateformationduringcýokinesisas we||as in celle|ongationcou|d be found in a form of various shapes of cells, organs and whole plants.Equal and inequal cell division,cell elongationand isodiametriccell growth- they all allow establishing,formingand maintainingcharacterof tissues,organsand whole bodies duringplantdevelopment. Scientificresearchhas alwaysbeenfocusedon regulatorycompounds,the actionof which induces detectable and observable responses. By uncovering the exact mechanisms of their mode of action, many intermediatemessengers were found. However,classicalconceptof "majorregulators'surviveduntilthesedays.lndeed,plant hormones, small organic molecules with wide range of effects, are typical representativesof such compounds,which are studiedalready around one hundred years.They are synthesisedand utilisedin varioustissuesthroughoutthe wholeplant and theycouldbe transportedin vasculatureto longdistances. Auxins and cýokinins - regu|atorsof plant development The researchof recentyears contributedremarkablyto the understandingof...
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