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Differentiation in Saccharomyces cerevisiae colonies
Čáp, Michal ; Palková, Zdena (advisor) ; Pichová, Iva (referee) ; Demnerová, Kateřina (referee)
Yeast colonies are, alongside to bacterial biofilms, multicellular communities formed by unicellular microorganisms. These specific communities differ in many ways from populations cultivated in planktonic cultivations. Gradients of nutrients, metabolic by- products and other factors are formed and preserved within these structures, which provides a basis for cellular differentiation. Current literature concerning these issues with emphasis on yeast colonies and biofilms is summarised in the Introduction of this work. Section Results then describes my contribution to the knowledge on the differentiation of the colonies of Saccharomyces cerevisiae as a model system for studying microbial multicellular structures. Previously described horizontal differentiation, i.e. differentiation between colony centre and margin, is further characterised with respect to ammonia signalling and stress resistance. The importance of genes conferring the cell`s oxidative stress resistance in colony differentiation was studied and it was concluded that not stress resistance but rather metabolic and other changes promoted by ammonia signal are important for colony differentiation and survival. A new type of colony differentiation - the horizontal, i.e. differentiation between upper and lower part of the colony, is...
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Antagonistic regulation by global transcription factors Tup1p, and Cyc8p of Flo11 and Flo11 -dependent phenotypes in wild yeast
Nguyen Van, Phu ; Palková, Zdena (advisor) ; Heidingsfeld, Olga (referee) ; Malcová, Ivana (referee) ; Demnerová, Kateřina (referee)
Biofilms are a common mode of yeast growth in which cells adhere to each other and adhere to biotic and abiotic surfaces to form complex multicellular structures. Living together in biofilms provides cells with several benefits, compared to planktonic cells such as protection and resistance to antimicrobials, environmental stresses and host immune attacks. Biofilms may play many important roles in commercial industries. But they are considered to be extremely dangerous in clinical settings. There is thus great interest in studying biofilms and how to eliminate them. In this study, we used wild yeast Saccharomyces cerevisiae colony biofilm as an ideal system to investigate potential functions of the yeast Cyc8p-Tup1p transcriptional corepressor complex in the regulation of yeast adhesion and biofilm formation on agar and at solid-liquid interfaces. Unexpectedly, we found that Cyc8p and Tup1p antagonistically control FLO11 expression and the formation of structured biofilm colonies on agar. Cyc8p itself acts as a key repressor of FLO11 and biofilm colony formation, whereas Tup1p promotes the formation of biofilm colonies and induces FLO11 expression by inhibiting the repressive function of Cyc8p and preventing Flo11p degradation possibly by inhibiting an extracellular protease. Other typical features...
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The Role of Microbiota in the Pathogenesis of Psoriasis
Stehlíková, Zuzana ; Jirásková Zákostelská, Zuzana (advisor) ; Demnerová, Kateřina (referee) ; Hrdý, Jiří (referee)
Psoriasis is a chronic, immune-mediated inflammatory skin disease. Its pathogenesis is associated with dysregulated cooperation among keratinocytes, innate and adaptive immune cells, coupled with environmental triggers, including microbiota. The aim of our study was to describe the microbiota composition in psoriasis and explore the role of bacteria and fungi in the pathogenesis of this disease. We used a mouse model of psoriasis induced by topical application of imiquimod (IISI) in both germ-free (GF) mice and conventional (CV) mice with microbiota manipulated by administration of a mixture of broad-spectrum antibiotics (ATB). ATB treatment markedly changed the intestinal but not the skin bacterial diversity and led to higher resistance to IISI in CV mice. Metronidazole was the most effective antibiotic, alleviating IISI symptoms in CV, but not in GF mice. This confirms that the effect of metronidazole on IISI was microbiota- dependent. Additionally, we characterized the microbiota composition of psoriatic lesions and unaffected skin in psoriatic patients compared to healthy controls, as well as the impact of different sampling approaches on uncovering cutaneous microbiota composition. We observed significant differences in α- and β-diversities when comparing identical samples sequenced on V1V2...
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Metodika odběru a izolace bakterií, kvasinek a plísní z fotografických materiálů
Demnerová, Kateřina ; Kadavá, Jana ; Purkrtová, Sabina ; Savická, Dana ; Sýkorová, Hana ; Ďurovič, Michal ; Benetková, Barbora ; Hricková, Kateřina ; Koukalová, Lucie ; Nováková, Martina
Metodika odběru a izolace bakterií, kvasinek a plísní z fotografických materiálů navrhuje optimální odběrový materiál a techniku odběru vzorku pro mikrobiologickou analýzu fotografických materiálů, která by v první řadě nepoškodila jejich povrch citlivý především na mechanické poškození, a zároveň vykazuje dostatečný záchyt mikroorganismů z povrchu. Je určena pro konzervátory, restaurátory a výzkumné pracovníky, kteří se věnují preventivní péči o fotografické sbírky v archivech, knihovnách, muzeích a galeriích.
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Soil microbial communities in agroecosystems and natural habitats contributing to resistance and resilience of the soil environment
Sarikhani, Ensyeh ; Kopecký, Jan (advisor) ; Demnerová, Kateřina (referee) ; Větrovský, Tomáš (referee)
Ensyeh Sarikhani Soil microbial communities in agroecosystems and natural habitats contributing to resistance and resilience of the soil environment. Summary The control of common scab of potatoes (CS) includes resistant varieties (cultivars), precise fertilization, increase of soil moisture, and chemical treatments. Yet, these management practices do not have common or reproducible results at differing sites. A monitoring study was done in 32 sites to evaluate the relation between CS and biological/chemical soil parameters. Correlations were observed between scab severity and content of nutrients such as Fe, N, and Ca in soil and periderm, and between disease severity and abundance of actinobacteria and total bacteria, together with the pathogenicity determinant, txtB gene (biosynthetic gene of thaxtomin) in both soil and periderm of potatoes. The findings led to novel conclusions, which can help to understand relationships applicable in scab control. Peat and DTPA chelated iron were supplemented to pots filled with soil conducive for CS in order to determine the effects of soil organic matter, iron and pH on CS development. The results were compared with data obtained for a suppressive soil from a nearby field with naturally low CS severity. Both peat and iron supplements decreased CS and the combination...
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The genetic basis of Czech garlic "paličák" (A. sativum L. ssp. ophioscorodon) focusing on the biosynthetic pathways of the secondary metabolites
Čermák, Vladimír ; Ovesná, Jaroslava (advisor) ; Demnerová, Kateřina (referee)
Czech bolting garlic (Allium sativum L.) is the name for Czech varieties, which are categorized into subspecies sativum ssp. An inflorescence production and specific onion morphology is typical for this type of garlic. The genetic basis, that has been described by the analysis of microsatellites, are discussed in this thesis. Transcription analysis has revealed a large number of unigenes that have been assigned by genetic ontology to individual functions in the organism. Therefore, genetic differences from other varieties were confirmed. The impact of the environment and cultivation practices, including large- scale production, is being explored. The results can be used for further research or breeding. Consumers and experts have rated the Czech bolting garlic as a variety with a distinctly pungent taste and aroma. Substances causing this strong characteristic taste are secondary metabolites, alk(en)ylcysteine-S-oxides (ACSO), especially alliin and methiin. Chemical analysis did not confirm the exceptionally high value of these taste precursors or other substances that should distinguish Czech bolting garlic. Secondary metabolism includes other important substances that are used mainly in pharmacy and healthcare, where garlic has been used for thousands of years due to antibacterial effects,...
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Study of biofilm formation of bacterial isolates from milk and dairy products
Nováková, Veronika ; Demnerová, Kateřina (advisor) ; Seydlová, Gabriela (referee)
Bacterial biofilm is a complex community of microbial cells, which are embed into the extracellular pollysacharide matrix. Typical ability of the biofilm is the ability to adhere to either abiotic, or biotic surfaces. The formation of biofilm is a dynamic process, which finally formes a 3D multicellular complex. The initial phase is called the adhesion. After that process is done, the extracellular polymeric matrix is produced to create living conditions of bacteria in biofilm. Biofilm has hight level of antimicrobial resistance. This resistance consists of physical and chemical barriers, which effectively block diffusion and penetration of antimicrobial substances inside the biofilm. Biofilms cause problems in food industry, where it could be origin of food contaminations. Thefore it is necessary to understand the relationship between microorganisms and materials used in food industry. Key words: biofilm, milk and dairy products, milk and dairy industry, disifectanc, Escherichia coli, Staphylococcus spp.
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Differentiation in Saccharomyces cerevisiae colonies
Čáp, Michal ; Palková, Zdena (advisor) ; Pichová, Iva (referee) ; Demnerová, Kateřina (referee)
Yeast colonies are, alongside to bacterial biofilms, multicellular communities formed by unicellular microorganisms. These specific communities differ in many ways from populations cultivated in planktonic cultivations. Gradients of nutrients, metabolic by- products and other factors are formed and preserved within these structures, which provides a basis for cellular differentiation. Current literature concerning these issues with emphasis on yeast colonies and biofilms is summarised in the Introduction of this work. Section Results then describes my contribution to the knowledge on the differentiation of the colonies of Saccharomyces cerevisiae as a model system for studying microbial multicellular structures. Previously described horizontal differentiation, i.e. differentiation between colony centre and margin, is further characterised with respect to ammonia signalling and stress resistance. The importance of genes conferring the cell`s oxidative stress resistance in colony differentiation was studied and it was concluded that not stress resistance but rather metabolic and other changes promoted by ammonia signal are important for colony differentiation and survival. A new type of colony differentiation - the horizontal, i.e. differentiation between upper and lower part of the colony, is...
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Differentiation of yeast colonies and development of new approaches to monitor oxygen and nutrient availability
Vopálenská, Irena ; Janderová, Blanka (advisor) ; Demnerová, Kateřina (referee) ; Pichová, Iva (referee)
Yeast Saccharomyces cerevisiae as an unicellular organism is one of the best-studied experimental organisms. It is an important model organism for the study of intracellular processes of eukaryotic cells. Yeasts are also social organisms with cell-to-cell communication able to form organized multicellular structures (colonies and biofilms). Yeast and other microorganisms in nature prefer to form colonies on solid substrates rather than to grow as "planktonic" single cells (Palková, 2004; Wimpenny, 2009). The yeast S. cerevisiae typically forms colonies, biofilms were described only rarely. Yeast colonies exhibit an organized morphological pattern characteristic of each particular yeast strain (Kocková-Kratochvílová, 1982). This work is focusing on morphology and differentiation of the S. cerevisiae colonies of common laboratory strains forming less structured colonies, and strains of the Σ1278b genetic background forming highly structured "fluffy" colonies. It shows that polarized budding pattern and especially cell ability to form aggregates enable development of structured morphology. During development of "fluffy" colonies two differently regulated events of dimorphic switch from yeast form to filamentous growth occur. One of these events is dependent on the surface glycoprotein, Flo11p flocculin. This...
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The Introduction of Contact Ecotoxicity Tests for the Assessment of Terrestrial Ecosystems
Modlitbová, Pavlína ; Kráčmar, Stanislav (referee) ; Beklová, Miroslava (referee) ; Demnerová, Kateřina (referee) ; Vávrová, Milada (advisor)
This dissertation thesis is focus on using toxicity tests with testing organism - terrestrial isopod Porcellio scaber. This organism is use for toxicity assessment of selected anorganic compunds which could contaminate terrestrial ecosystem. This organism was selected for several reasons - sophisticated test methodology; well known biology of these animals and their ecological relevance. Besides classical endpoints such as mortality, change in mass of test specimens and the effect of the presence and concentration of contaminants onto food consumption; we monitored changes in the morphometric characteristics at the level of tissues and cells, the cytotoxicity and possibly bioaccumulation in various parts of the body of the organism. Selected test substance in this thesis were nanoparticles and inorganic salts. Wherein for chosen gold nanoparticles was carried out a complete study presents several levels of organization of living matter (organism, tissue, cell). For selected salts compound, this study was carried out on the behavior of organisms and classical endpoints.
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