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Structure and function of bacterial communities during succession on dead plant biomass
Tláskal, Vojtěch ; Baldrian, Petr (advisor) ; Marečková, Markéta (referee)
The decomposition of dead plant biomass substantially contributes to the carbon cycle and therefore is one of the key processes in temperate forests. While the role of fungi in litter and deadwood decomposition was repeatedly addressed, there are just a few surveys of bacteria associated with decomposing plant biomass. The development of bacterial community within leaf litter is likely driven by the changes in litter chemistry and by the availability of nutrients in the litter. Fungal activity greatly contributes to changing properties of substrate and thus influences bacterial community. Availability of nutrients is changing during biomass decomposition from easily accessible substrates toward more recalcitrant ones (e.g. lignin). The colonization of deadwood by bacteria is influenced by various factors such as microclimate conditions, tree species and volume. The aim of this thesis was to describe bacterial community dynamics during the first two years of decomposition of leaf litter and deadwood. In the leaf litter experiment, bacterial community was analysed in the live, senescent and decomposing leaves of Quercus petraea. This experiment was performed in the Xaverovsky Haj Natural Reserve, Czech Republic. Deadwood experiment was focused on the composition of bacterial community in the initial...
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Vegetation of post-mining sites determines soil microbial community structure and soil processes
Urbanová, Michaela ; Baldrian, Petr (advisor) ; Bárta, Jiří (referee) ; Chroňáková, Alica (referee)
Vegetation of post-mining sites determines soil microbial community structure and soil processes Mgr. Michaela Urbanová Abstract The aim of this thesis, which consists of four published articles, was to investigate the effect of vegetation on soil microbial communities and processes in de novo developing soil substrate on the brown-coal spoil heaps in the surrounding of city Sokolov. Spoil material - soil clayey substrate, which had been gradually mined from the opencast brown coal mine, stratified onto spoil heaps and reclaimed by assisted afforestation with selected tree species or left for spontaneous plant succession, changes its biotic and abiotic characteristic in the course of time and particularly under the influence of plants. Changes of spoil substrate characteristics are related to the growth of plant roots and particularly also to the production of plant biomass, which is decomposed gradually and takes part of soil, where participates to soil organic matter. The process of plant dead materials decomposition and transformation is the function of the activity of soil organisms and among them notably soil microorganisms. Moreover, the presence of many of them is closely related to the presence of vegetation, whose symbionts or pathogens are. The exact mechanisms of the plant-microbes interactions...
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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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Change of structure and function of a fungal comunnities in the soil environment after the end of the flowassimilates with Norway spruce (Picea abies)
Charvátová, Markéta ; Baldrian, Petr (advisor) ; Mrnka, Libor (referee)
The decomposition of plant biomass is associated with changes of the associated fungal community which is crucial in the process of degradation due to their production of extracellular enzymes. Moreover, fungal communities as well as enzymatic activity are specific for soil microhabitats. This thesis focused on the importance of fungi associated with the roots of Norway spruce (Picea abies) trees. Forest with dominating Norway spruce, located in the School Forest Enterprise Křtiny area in Moravia, was selected for the experiment. The aim of this thesis was to compare the community composition of fungi in the roots, rhizosphere and bulk soil and to describe the gradual development of this community after the death of the roots. The next aim was to isolate, describe and characterize those fungi that decomposed roots 5 and 9 months after tree harvest. Identification of fungal isolates was based on DNA sequencing. Degradation capabilities were determined as the activity of enzymes involved in the degradation of lignocellulose. Composition of fungal communities was described by sequencing of environmental DNA on the Illumina platform. Before the flow of assimilates from the live trees stopped, communities of ectomycorrhizal fungi dominated with 60 % of all sequences. Increased abundance of saprotrophic...
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Fungi associated with decomposing wood in temperate forests
Štercová, Lucie ; Baldrian, Petr (advisor) ; Zikánová, Blanka (referee)
Wood decaying fungi are an essential part of all forest ecosystems. On their functioning depends a number of other organisms which use substances produced during decay of wood. Enzymes that degrade wood represent indispensable tool for fungi in converting structural compounds of wood to water and carbon dioxide. That makes them an essential part of the carbon cycle in nature. The dynamic of fungal communities on decaying wood is determined by a range of abiotic and biotic factors. The variability of microclimatic conditions, differences in the wood humidity and a gaseous mode are identifying stress factors which define the presence of species adapted to those conditions. An important factor, with which we can partially predict community composition, is the degree of decomposition of wood and the history of a species on the substrate. Another indicator, which may partly explain the composition of the community is a way of dying of a tree and a type of decaying substrate. Often, the development of a community follows from the primary colonizers, with high tolerance to unfavorable conditions, through the secondary colonizers, which have the ability to obtain the substrate over primary colonizers, but require more stable microclimatic conditions, to late colonizers, who are adapted to stress factors as...
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Cellulolytic fungi and their diversity on plant litter
Gálová, Diana ; Baldrian, Petr (advisor) ; Kolařík, Miroslav (referee)
Litter decomposition requires the presence of corresponding degradative enzymes, produced mainly by fungi. Forest soils show considerable spatial heterogeneity of distribution of these enzymes at diferent scales. Moreover, enzyme pruduction varies during the year, usually accompanied by the change in fungal community composition. In this work I examined if this spatial heterogeneity can be seen even at a scale of an individual leaf and whether the fungal community differs among enzyme activity hotspots and inactive parts of the leaves. Another goal was isolation of celulytic fungi from cellulose litterbags incubated on forest floor using particle filtration and dilution-to-extinction method. In a broadleaved forest dominated by oak leaves at different stages of decay were collected: senescent leaves on twigs, and leaves after 2, 10 and 22 months of decomposition. Ten leaves per season were taken for analysis of cellobiohydrolase activity over the leaf surface. Leaves were attachmed onto melted agarose plate and leaf surface was covered with low melting point agarose containing fluorescently labelled substrate. For each leaf a map of enzyme activity was created and area with the high and low enzyme activity was identified. From both sites a square of approx. 1 cm2 was cut out, DNA was extracted and fungal...
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Ecophysiology of microscopic fungus Pseudogymnoascus destructans
Homutová, Karolína ; Kubátová, Alena (advisor) ; Baldrian, Petr (referee)
A microscopic fungus Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) causes illness known as white-nose syndrome (WNS) causing death of bats during hibernation. The illness occurs in the North America and in Europe. The fungus is characteristic by asymmetrically curved conidia, by slow growth and growth at low temperatures (below 20 řC). The aim of this study is to clarify properties responsible for unique ecelogy of Pseudogymnoascus destructans by comparison with ecological related or unrelated pathogenic or nonpathogenic fungi. This part includes study of tolerance to physiological stresses and recognition of spectrum of utilizating nutrients (compounds of carbon, nitrogen, phosphorus, sulphur and nutrient supplements). Testing to physiological stresses should help to estimate a potentiality of fungus to spread out of caves. The last aim is to develop a selective isolation medium for P. destructans. Influence of several types of physiological stress (e.g. UVA, UVA with UVB, 25 řC, 30 řC, 37 řC and dryness) was investigated with fluorescent stain propidium iodide (PI) by flow cytometry. The spores of Pseudogymnoascus destructans and three fungi from underground spaces were not viable after 3 weeks at 37 řC. Other stresses did not cause a decreasing of viability or some stresses caused...
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Ecology of deadwood-associated fungi in the ecosystems of nature-like forests
Zrůstová, Petra ; Baldrian, Petr (advisor) ; Konopásek, Ivo (referee)
Dead wood plays an important role in forest ecosystems in the context of C dynamics, nutrient cycling, forest regeneration and biodiversity. Decaying wood sustains biodiversity by providing habitats and energy for fungi, bacteria, invertebrates, and many other organisms. Dead wood is resistant to decomposition and its decay is driven mainly by filamentous fungi. Community structure of wood- inhabiting fungi changes during decomposition, but the relationship between substrate quality and decomposer community is still poorly understood. This work studied fungal community composition with respect to tree species, stage of decay, volume and physico-chemical properties (such as pH, carbon and nitrogen content) of dead wood. Fungi were identified using next generation sequencing approaches - 454-pyrosequencing and Illumina MiSeq sequencing. Tree species, volume of dead wood (branches x logs) and stage of decay were the main variables affecting fungal community composition. Higher enzyme activities and content of fungal biomass indicate faster colonization of small branches than tree trunks by fungi. Fungal community composition, wood chemical properties and enzyme activities changed during decomposition. Both content of nitrogen and fungal biomass increased during decomposition. Enzyme activites peaked...
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Molecular biology of soil fungi participating in litter decomposition in forest ecosystems
Voříšková, Jana ; Baldrian, Petr (advisor) ; Gryndler, Milan (referee) ; Koukol, Ondřej (referee)
In forest ecosystems, substantial part of carbon enters soil in the form of plant litter. The decomposition of litter and soil organic matter represents an important process affecting nutrient cycling and carbon balance in soils. Fungi are considered the primary decomposers in terrestrial ecosystems due to the production of wide range of extracellular enzymes that allow them to attack the lignocellulose matrix in litter. Even if fungi represent key players in organic matter decomposition, the information about the structure and diversity of their communities is still limited and the roles of individual fungal taxa in forest soils remain unclear. This Ph.D. thesis focused on the characterization of fungal communities in forest soils and their potential to decompose plant litter. The method for in-depth analysis of complex microbial communities from environmental samples was established and used. In addition, single eukaryotic functional gene was analysed in soil for the first time at a depth that allowed reliable estimation of diversity. It was demonstrated that microbial community composition differs among horizons of forest soil profile. Despite similar diversity, significant differences in microbial community composition were observed between the DNA and RNA. Several microbial groups highly...
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Ecological roles of bacteria associated with plant litter
Tláskal, Vojtěch ; Baldrian, Petr (advisor) ; Seydlová, Gabriela (referee)
Leaf litter in temperate forests represents an important input of carbon into the soil. Main players in the decomposition of leaf litter are fungi and bacteria. While the role of fungi in litter decomposition was repeatedly addressed, there are just a few field studies where litter-associated bacteria were also considered. The aim of this work was to summarize available literature studying the structure and function of bacterial community during litter degradation. The nature of different possible substrates is discussed. Genetic approach is briefly outlined. Factors such as diversity, spatially distribution and abiotic factors that can influence community are also considered. It can be supposed that community composition changes with the change of litter chemistry and nutrient availability. The development of bacterial community might be driven by the decreasing availability of nutrients in litter. Labile compounds are utilised at the beginning of decomposition, while recalcitrant substrates are utilised later by specialist taxa. Members of the phyla Actinobacteria, Acidobacteria, Bacteroidetes and Proteobacteria seem to be dominant during decomposition. Different types of relationships between fungi and bacteria also most likely influence the composition of community. This review shows that the bacterial...
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