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Combination of biochemical and high-throughput-sequencing approaches to study the role of Antinobacteria and fungi in the decomposition of plant biomass
Větrovský, Tomáš ; Baldrian, Petr (advisor) ; Slaninová Kyselková, Martina (referee) ; Tomšovský, Michal (referee)
Dead plant biomass is a key pool of carbon in terrestrial ecosystems. Its decomposition in soil environments is thus an essential process of the carbon cycle. Fungi are considered to be the primary decomposers in soil ecosystems because of their physiological adaptations and enzymatic apparatus composed from highly effective oxidative and hydrolytic enzymes. Many recent works show that in addition to fungi, bacteria may also play a significant role in lignocellulose decomposition and among bacteria, the members of the phylum Actinobacteria are often regarded to significantly contribute to cellulose and lignocellulose decomposition. This thesis is focused on the evaluation of the role that fungi and Actinobacteria play in dead plant biomass degradation. First, it explored mechanisms involved in degradation, in particular the enzymatic breakdown of major lignocellulose components as cellulose, hemicelluloses and lignin. Enzymatic apparatus of the saprotrophic fungus Fomes fomentarius was explored both in vitro as well as in vivo. Several Actinobacteria were isolated from soil and comparative experiments, investigating production of hydrolytic enzymes, were carried out to track the transformation of polysaccharides and lignin by these strains. To explain the roles of lignocellulose decomposers in...
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Obesity monitoring in middle aged men in Hradec Králové
Senecký, Petr ; Bunc, Václav (advisor) ; Větrovský, Tomáš (referee)
Title: Obesity monitoring in middle aged men in Hradec Kralove Targets: The targets of thesis is to determine prevalence of the obesity in men at age 30 - 50 years in Hradec Kralove. The necessary data for this empirical research will be obtained on the basis of the questionnaires distributed among 30 active athletes and 30 pacients of prof. Martinik's diabetology office, who suffer from obesity and undergo treatment in his office. Subsequently, I will perform a deep analysis of all the data obtained from the questionnaries, in order to identify hazard factors for obesity, stress management, physical aktivity, fixed daily routine and life management or the prevalence of the genetic load in the group of surveyed athletes and surveyed obese patients of prof. Martiník's diabetology office. These data will be then compared in order to identifily the main differences between active athletes and obese patients. Methods: The empirical research was conducted at 30 randomly selected active athletes (at age 30 - 50 years), who live in Hradec Kralove and at 30 random patients (also at age 30 - 50 years) of prof. Martiník, who also live in Hradec Kralove and undergo medical treatment on the basis of the questionnaire, which I created myself and filled personally with the patients and athletes in order to...
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Pyrosequencing analysis of fungal assemblages and the effect of ITS rDNA intragenomic variation on diversity estimates
Zelenka, Tomáš ; Kolařík, Miroslav (advisor) ; Větrovský, Tomáš (referee)
The ITS region of nuclear ribosomal DNA has recently become a frequently studied region for its use as a barcode marker. It is employed in environmental metagenomic analyses, the determination of fungi and in studies of fungal phylogenetics. In genomes, rDNA occurs in the form of large multicopy tandem arrays. Thus common Sanger sequencing leads to a consensus of many copies which in fact conceals most of the potential intragenomic variation. However 454 pyrosequencing reveals the sequence of single copies. Although it is believed that most of the variation among copies is reduced during a process called concerted evolution, some variation might still be conserved, including variation of non-functional pseudogenes. As a consequence, when using 454 pyrosequencing, we are not able to discriminate between this variation and real diversity. This could have a huge impact on the estimation of real diversity as well as on the correct assessment of pyrosequence studies. This thesis reviews current knowledge of intragenomic variation among fungi and summarizes some papers applying pyrosequencing in the research of fungal diversity. At the same time it indicates intragenomic variation as a potential cause of untrue diversity in diversity studies. Up until now there hasn't existed any experimental survey covering...
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