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Morfogenéza epidermálnych buniek listov Arabidopsis thaliana v odpovedi na chladový stres a zníženú dostupnosť svetla
Tekulová, Veronika
The bachelor thesis was focused on the physiological, cellular and transcriptomic characterization of Arabidopsis thaliana treated with cold under different light conditions. To determine the role of the cold stress in leaf ontogenesis, the analysis included leaves in the different developmental stages. It was found that 4 weeks old plants have almost completely developed 6th leaf and cold treatment has no impact on its cellular morphology. In contrast, younger leaf number 14 which initiates a transition from proliferation to expansion stage was strongly affected by cold treatment. It is shown that exposition to a low temperature significantly reduces its cell area and a number of epidermal cells. Propidium staining confirmed that young proliferating leaves are also more sensitive to stress conditions than older leaves. The higher sensitivity of young leaves to cold stress was accompanied with lower expression of CBF genes that play an important role in cold stress tolerance. Younger leaves also showed higher expression of cytokinin responsive genes and genes related to cell cycle corresponding to ongoing developmental processes.
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Odpoveď ekotypov Arabidopsis na abiotický stres
Zvalová, Monika
Distribution of a given plant species reflects its ability to acclimate to different natural environmental conditions. Plant acclimation to cold is of high importance because of recent raise of abundance of temperature extremes. The aim of this thesis was a characterization of twentytwo selected Arabidopsis ecotypes after sub-zero treatment and identification of those that are associated with effective cold acclimation. This evaluation of the processes involved in cold protection was enabled by substantial variability in the frost tolerance among these contrasting ecotypes. We identified frost tolerant and sensitive ecotypes that were further characterized by rosette area and photosynthetic pigments measurement. These results correspond with geographical distribution of a given species. The next step will be proteomic analysis of selected ecotypes to identified molecular processess involved in freezing stress tolerance. The adaptation of individual Arabidopsis ecotypes to different environmental conditions, resulting in a wide range of frost hardiness, offers unique possibility to elucidate mechanisms associated with efficient defence to low temperatures.Formula clause:I agree that my work will be published in accordance with § 47b of Act no. 111/1998 Coll., On Higher Education Institutions and on Amendments to Other Acts (Higher Education Act), as amended, and in accordance with the valid Directive on the Publishing of Higher Education Final Works
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Role of sequence context in DNA methylation
Polák, Jan ; Fischer, Lukáš (advisor) ; Širl, Marek (referee)
Cytosine methylation of DNA is a pivotal epigenetic mark, which contributes to the regulation of the gene expresion, silencing of transposable elements, and co-defines chromatine state. There are three cytosine contexts: CG, CHG and CHH (where H stands for C, A, or T). Arabidopsis thaliana (and plants in general) has an arsenal of molecular mechanisms capable of cytosine methylation in all of its contexts. That said, there are two tasks at hand: maintaining of pre-existing methylation and if need be, creating new methylated spots. The actual process of maintaining of the methylation depends on the cytosine context. Methylation of symmetrical contexts of CG and CHG can utilize the information about the methylation pattern from the second DNA strand. The aymmetrical context of CHH, and also CHG need to look for this information elsewhere: in the methylation of the lysine 9 of H3 histone. This creates a self-reinforcing loop and a crosstalk between two epigenetic mechanisms. Maintaince of methylation of CHH is also navigated by small RNA complementary to the locus in question. This mechanism of enzyme navigating by RNA is also used in establishing a new methylated site for all of the contexts. CG methylation is most prevalent in both heterochromatine and euchromatine. It also has a special functions...
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Molecular basis for regulation of cell wall pH in Arabidopsis thaliana
Bogdan, Michal ; Fendrych, Matyáš (advisor) ; Retzer, Katarzyna (referee)
The cell wall is one of the defining parts of the plant cell. Cell walls resist turgor pressure, give plants their rigidity and still allow the cell to grow despite their stiffness. The plant cell wall is a composite material that consists of different elements; thus, the knowledge of the plant cell wall composition helps us understand the impact of the mentioned pH changes. Alterations of plant cell wall properties result in a change of plant cell growth rate. A prime example of this is the modification of wall properties by pH change, termed the acid growth. Although acid growth has been studied for a long time, we still do not fully understand the underlying mechanisms, as we have not yet identified all the agents involved in acid growth and our ability to determine apoplastic pH is limited. Local cell wall pH fluctuations arise also due to plant organs having different roles and being affected by a diverse range of stimuli. Despite the fact that new pH measurement techniques, such as genetically encoded fluorescent probes, were developed in the last two decades, there is still a need to provide a higher spatiotemporal resolution of pH analysis. Key words: Arabidopsis, apoplastic pH, cell growth, acid growth theory, cell wall, auxin, apoplastic pH measurement
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Mechanisms of establishment and maintenance of PIN polarity in Arabidopsis
Glanc, Matouš ; Friml, Jiří (advisor) ; Grebe, Markus (referee) ; Luschnig, Christian (referee)
Cell polarity is a key concept in plant biology. The subcellular localization of Pin- formed (PIN) auxin efflux carriers in the root of "#$%&'()*&* is remarkably asymmetrical, making PINs prominent markers to study cell polarity. In spite of its developmental importance and two decades of research, the molecular basis of PIN polarity remains largely unknown. In this thesis, I employed advanced transgenic and fluorescence microscopy approaches to gain insight into several aspects of PIN polarity regulation. I participated in establishing a novel genetically encoded inhibitor of endocytosis, an invaluable tool for the study of the importance of endocytosis for various cellular processes, including PIN polarity. I demonstrated that apical polarity of PIN2 needs to be re-established after cell division and that this process depends on endocytosis, '+!,(-( protein secretion and the action of WAG1 and related protein kinases, but not transcytosis, cell-cell signaling or intact cytoskeleton. Finally, I identified the previously unknown role of MAB4/MEL proteins in PIN polarity, which lies in the ability of MAB4/MELs to reduce PIN lateral diffusion and thus contribute to PIN polarity maintenance. My results, besides broadening current understanding of PIN polarity regulation, identify mechanisms that...
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Altitudinal adaptation of Arabidopsis
Požárová, Doubravka ; Kolář, Filip (advisor) ; Pinc, Jan (referee)
Study of adaptation provides insights into a key part of evolutionary processes. This thesis presents an overview of an adaptation process and further focuses on local adaptation - a process of selection of phenotypes and underlying genotypes that fits the best to a particular environmental condition. Then, the thesis provides a brief summary of methodical approaches how to detect local adaptation, on both, molecular and phenotypic level. Further I focus on alpine environment as a model to study local adaptation, due to complex set of stresses imposing strong selection pressure, their variable strength structured along altitudinal gradient and island-like distribution triggering parallel origins of locally adapted ecotypes. Such is also the case of local adaptation to mountain environment in plant model genus Arabidopsis. I here review the responses to environmental variables along altitudinal gradients and synthesize the environmental factors that are involved in the local adaptation process: precipitation, water regime, temperature and radiation. The recorded adaptive traits were associated mainly with life history, phenology, growth forms and drought stress tolerance. Study of alpine adaptation helps with understanding evolutionary and ecological processes and it is also important to assess how...
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Introgressive zone of Arabidopsis lyrata and A. arenosa in Central Europe
Hojka, Jakub ; Marhold, Karol (advisor) ; Španiel, Stanislav (referee)
Genetic composition of hybrid zones often reflects geographical and ecological gradients. Hybrid zone of Arabidopsis lyrata and A. arenosa in Central Europe can be convenient model system for testing such hypothesis. These are well circumscribed but still interfertile, and their offspring is fully fertile. Besides repeated hybridisation events, the current state is complicated by the autopolyploidisation events of one of the parents, namely A. lyrata, which is present in the area studied both as diploid and tetraploid. This hybrid zone was already described in previous publications, which showed certain gradient of introgression. Moreover, the area of the hybrid zone represents a transition between Alpine and Pannonian climate along an altitudinal gradient leading from the Prealps to the lowland Wienerwald. The current thesis is the as yet most detailed study of this hybrid zone both in respect of number of studied populations and the amount of data acquired using the methods of RAD Sequencing, multivariate morphometrics and flow cytometry. The analysis of genetic data showed a gradient of introgression, where parental populations are placed at opposite ends, whereas genetically intermediate hybrids are placed in its centre. Analysis of the genome size showed an additional gradient, where hybrids...
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Characterization of the exocyst complex SEC15 subunit in A. thaliana
Aldorfová, Klára ; Hála, Michal (advisor) ; Hafidh, Said (referee)
The final step of secretion termed exocytosis is mediated by the exocyst complex. The exocyst is an evolutionary conserved protein complex that tethers secretory vesicle to the target membrane and consists of eight subunits: Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo84, and Exo70. Sec15 exocyst subunit was previously shown to connect the rest of the exocyst complex with a secretory vesicle in yeast, mammals and fruit fly via interaction with Rab GTPase and GEF of Rab GTPase. Here, I show that plant SEC15B potentially functions in evolutionary conserved manner. First, two mutant lines of Arabidopsis thaliana sec15b mutant were tested in characteristics typical for other exocyst mutants. Although some characteristics reach certain level of plasticity, both sec15b-1 and sec15b-2 show similar tendencies, which are mostly consistent with defects with other mutants in exocyst subunits. sec15b-1 has been determined as a stronger allele that is defective in formation of seed coat, elongation of etiolated hypocotyl, growth of stem and primary root, establishment of axillary branches and lateral roots, diameter of rosette and, unexpectedly, growth of pollen tubes. Phenotype of sec15b-1 was rescued by insertion of SEC15B gene under SEC15B promotor. Second, complementation test showed that SEC15B and SEC15A are...
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Functions of Exocyst Complex in the Regulation of Stomata Dynamics
Röder, Matěj ; Žárský, Viktor (advisor) ; Burketová, Lenka (referee)
Stomata are structures in plant epidermis which regulate contact between inner and outer environment of the plant by mediating their stomatal aperture. Many inner and outer signals contribute to the ontogenesis of the stomatal pattern. Guard cells undergo significant change of volume and surface during stomatal movement. This change of surface must be compensated by intracellular trafficking of membrane material because biological membrane has limited elasticity. Most of this trafficking takes place between plasma membrane and endosomal compartments. Complex exocyst is protein complex that ensures proper targeting of secretory vesicles to their destination on the plasma membrane. Function of this complex is essential for many cellular processes that require precise targeting of secretion. Mutation in gene Exo70B1 causes different development of the stomatal pattern. Plants with mutated Exo70B1 differ in stomatal size depending on the cultivation conditions more than wild type plant. Protein EXO70B1 is also directly involved in stomatal dynamics because mutants exo70B1 have retarded stomatal opening in response to light. This direct connection can be observed on the fluorescently labeled protein EXOB1 which significantly changes its localization during stomatal movements. None of these observed phenotypes is...
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Is there any influence of ecological gradient on the introgression and its evolutionary consequences in the genus Arabidopsis?
Hojka, Jakub ; Marhold, Karol (advisor) ; Mráz, Patrik (referee)
1 Hybridization, introgression and polyploidy, especially in their combination, belong to the most important speciation processes in vascular plants. Athough basic features of these processes are generally well known, their cases in particular groups of plants and concrete cases differ a lot and their studies often bring interesting results. In the genus Arabidopsis several cases of combined hybrid and polyploid speciation are known (e.g. A. kamchatica, A. suecica) and there are also cases where we expect introgressive hybridization (e.g. between A. arenosa and A. petraea). The aim of the bachelor thesis is an evaluation of known cases of homo- and heteroploid introgressive hybridization and hybrid speciation in the genus Arabidopsis in the context of similar cases in the family Brassicaceae and perhaps also other families of vascular plants. Next aim is to design method of detailed evaluation of the introgressive zone of A. arenosa and A. petraea (= A. lyrata subsp. petraea) in Central Europe using mophological, cytometric and molecular evidence as well as ecological parameters of studied localities. Set of morphological characters will be tested on the currently available material of A. arenosa that is cultivated at CEITEC Brno as part of the finalised GACR project.
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