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Physiological changes of cereals under temperature stress
Kantová, Anežka ; Vítámvás, Pavel (advisor) ; Hnilička, František (referee)
Cereals are among the oldest crops that have been grown and used by humans as important component of their diet. It is an important source of livelihood for the human population and have a wide range of uses, mainly in the food industry. Cereals generally serve as a source of energy in the diet, due to the high starch content. The most commonly grown types of cereals are especially wheat, barley, rye, rice, corn, but there may be other species such as oats and millet. However, even cereals do not avoid the problems associated with the action of abiotic stress factors. Their effect on all plants is manifested by a decrease in vitality, but in crops - such as cereals - mainly by a decrease in yield. Due to the reduction in yield, breeding of resistant cereal genotypes is now in the primary interest of breeders. This work summarizes the basic principles of the action of heat/cold stress on plants and explains the reactions of various types of cereals to these abiotic stress factors. Key words: proteome, physiology, cereals, temperature stres, yield, abiotic stres
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Jak rostlinám chutná proteinová strava
Zronková, Veronika
Nitrogen is one of the most abundant elements in living organisms. In contrast to other key elements, its input from inorganic sources is limited and thus the most of the nitrogen pool has to be recycled from nitrogenous compounds (including proteins) that have been used by other organisms. This thesis How do plants enjoy protein nutrition briefly reviews nitrogen assimilation mechanisms and discusses alternative nitrogen sources for plants. The experimental part provides evidence that an intact protein in the culture medium decreases root growth and that the cytokinin signalling participates in this response. Then, to provide tools for further elucidations of this mechanism, a method for plant secretome analyses was tested. Altogether, more than 300 proteins were detected in hydroponic-based cultures of Nicotiana tabacum and more than one-third of these were affected by the cytokinin treatment.
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Srovnávací analýza působení exogenního působení cytokininů s inhibicí degradace jejich endogenních hladin u Arabidopsis thaliana
Koukalová, Vladěna
Agricultural production is negatively affected by stress factors like drought or rising ambient temperature. Genetic engineering and production of specific GMO seems to be very promising in protection of the crop yield however these are strictly limited for the use in the field. For this reason alternative approaches like application of growth regulators are in the focus of recent research. INCYDE is a new potent growth regulator discovered as inhibitor of cytokinin degradation. In this Diploma thesis we show that high levels of INCYDE could also activate AHK4, cytokinin receptor, and act like cytokinins. Comparison of trans-zeatin and INCYDE showed that application of these regulators results in spatial-specific pattern in activation of the cytokinin signaling. trans-zeatin and INCYDE also induce different changes in proteom mainly in proteasome pathway, gluconeogenesis and fatty acids metabolism. Application of INCYDE simultaneously with inducing drought stress revealed that INCYDE could decrease sensitivity to this stressor. This feature seems to be very perspective for its utilization in the agriculture.
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Posttranslační modifikace proteinů a jejich role v klíčení rostlin
Berka, Miroslav
Protein posttranslational modification represents a complex extension of proteome diversity and provides a transcription-independent control of cellular processes. This thesis "Protein posttranslational modifications and their role in seed germination" briefly reviews major posttranslational modifications, highlights the role of protein phosphorylation in plant regulatory circuits, and summarises enrichment techniques that are employed in phosphoproteomics. The experimental part describes the development and optimisation of methods that can be utilised in a monitoring of protein posttranslational modifications during barley (Hordeum vulgare L.) seed germination, including a tissue fractionation, proteome equalisation and peptide fractionation techniques. Finally, these methods were employed in an analysis of hydrogen peroxide effect on seed germination. In total, over 1,400 proteins were identified in individual fractions, quantitative profiles were established for more than 881 (~60%) and the validated differentially abundant proteins could indicate a potential mechanism behind the peroxide stimulatory effect. Peptides that would correspond to the identified seed phosphoproteins were mostly inaccessible in this experiment and would require optimised targeted analysis.
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Abiotické a biotické faktory ovlivňující klíčení rostlin
Berka, Miroslav
Seed germination and early plant development is a crucial phase of plants' life. Multiple internal and external stimuli influence germination progress and have a serious impact on a plant's survival and vitality. Biotic and abiotic stimuli trigger a whole range of changes, both on molecular and developmental levels, but the complex molecular mechanisms regulating these responses are far from being resolved. This thesis reviews the seed germination process and outlines the role of external stimuli in its progress. The experimental part describes the development of a method for seed germination monitoring, provides new insight into the role of hydrogen peroxide in germination, and analyzes effects of cadmium ions, temperature, salt and drought on proteome and metabolome of germinating seeds of Hordeum vulgare. In total, 2000 proteins and 800 metabolites were identified. The analyses revealed over 95 putative abiotic stress markers, including 63 and 36 proteins and metabolites, respectively
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Charakterizace rostlinného proteomu
Petrová, Silvie
Plant leaf tissue proteomics analyses face multiple challenges, including the presence of highly abundant proteins like RuBisCO. This thesis reviews methods in protein analysis and outlines proteome fraction techniques. The experimental part compares the leaf proteome composition of five plants, including three representatives of herbaceous (Arabidopsis thaliana, Solanum lycopersicum, Hordeum vulgare), and two woody plants (Malus domestica and Populus hybrid). Further, peptide fractionations by a high-pH C18 and SCX chromatography, and the role of data processing in proteomics analyses are illustrated.
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Proteom a metabolom parazitů rodu Phytophthora
Zelená, Pavla
Genus Phytophthora represents a world-wide spread pathogen with more than hundred recognized species and its devastating effect on plants has a serious economic and ecological impact. This diploma thesis entitled „Proteome and metabolome of genus Phytophthora” briefly summarizes knowledge about this pathogen, including its life cycle and interactions with its host. Twelve species representing six Phytophthora clades that were selected for experimental work are then discussed in details. Phytophthora isolates were characterized on proteome and metabolome level employing an LC-MS untargeted proteome profiling and a GC-MS analysis of volatiles. The results were then processed to identify candidate molecules for a targeted identification of Phytophthora and these results were validated in an independent experiment with P. palmivora and Hordeum vulgare. We found that a proteome profiling can be employed as a tool to differentiate individual Phytophthora species and that the marker peptides can be employed for a targeted monitoring of Phytophthora presence in plants.
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Morphological, physiological and proteomic changes of cereals under abiotic stress
Kantová, Anežka ; Vítámvás, Pavel (advisor) ; Hnilička, František (referee)
Cereals are among the oldest crops that have been grown and used by humans as important component of their diet. It is an important source of livelihood for the human population and have a wide range of uses, mainly in the food industry. Cereals generally serve as a source of energy in the diet, due to the high starch content. The most commonly grown types of cereals are especially wheat, barley, rye, rice, corn, but there may be other species such as oats and millet. However, even cereals do not avoid the problems associated with the action of abiotic stress factors. Their effect on all plants is manifested by a decrease in vitality, but in crops - such as cereals - mainly by a decrease in yield. Due to the reduction in yield, breeding of resistant cereal genotypes is now in the primary interest of breeders. This work summarizes the basic principles of the action of abiotic stress on plants and explains the reactions of various types of cereals to abiotic stress factors. Key words: proteome, physiology, cereals, abiotic stress, yield
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Amastigotes of various origins: comparison of proteome and development in a natural vector.
Pacáková, Lenka ; Leštinová, Tereza (advisor) ; Paris, Zdeněk (referee)
Amastigotes are forms of Leishmania, naturally occurring in vertebrate hosts within phagocytic cells - especially the macrophages. The aim of this project was to compare three types of amastigotes of Leishmania that can be used for experiments under laboratory conditions - namely the axenic amastigotes, cultured extracellularly (without vertebrate phagocytic cells), amastigotes isolated from macrophages infected ex vivo, and "true" amatigotes isolated from lesions of the infected BALB/c mice. Amastigotes were compared with respect to the development in the natural vector and at the proteome level. L. mexicana, the causative agent of cutaneous leishmaniasis in the New World, was chosen for this comparison. In experiments comparing the development of Leishmania in the natural vector Lu. longipalpis we found significantly weaker infections in the sand flies infected with axenic amastigotes compared to other types of amastigotes. In addition to the intensity of infection, we compared the localization of promastigotes in the digestive tract of the phlebotomine sand flies. The following localizations were observed: the abdomen, the thorax, the cardia and the stomodeal valve, which is crucial for infectivity of the sand fly. There was no significant difference in localization in any of the groups of...
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Characterization of unique proteins of Giardia intestinalis and their role in mitosomal biogenesis.
Zemanová, Tereza ; Doležal, Pavel (advisor) ; Rada, Petr (referee)
The unicellular parasite Giardia intestinalis is one of the organisms carrying mitochondrion-related organelle known as mitosome, which is adapted to the microaerobic lifestyle. The only known fuction of the mitosome is the synthesis of the iron-sulphur clusters. The research of the mitosomal proteome provides new information on the biogenesis and function of this unusual organelle. One of the means of the mitosome research is the analysis of the interactome of the known mitosomal proteins. The state-of- the-art method of the interactome approach is the use of the chemical crosslinking and the subsequent immunoaffinity isolation of the complexes, containing the protein of interest. In this thesis, the interactomes of GiTom40 and GiMOMP35 were characterized with the bioinformatic tools. The cellular localization of four of the chosen proteins was determined by the fluorescent microscopy. One of the proteins, the predicted dynein intermediate chain DIC6939, was phylogenetically classified as an axonemal dynein. The superresolution microsopy was utilized to observe the possible colocalization of DIC6939 with the mitosomes and blue native PAGE led to the visualization of its native complexes. In this work, the optimal conditions for DIC6939 interactome isolation were succesfully determined. The outcome...
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