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Bacterial interactions and secondary metabolites in plant and soil environment
Rapoport, Daria ; Marečková, Markéta (advisor) ; Lovecká, Petra (referee) ; Brabcová, Vendula (referee)
Phylum Actinobacteriota represents one of the major phyla in plant and soil environments. Known for the ability to produce numerous secondary metabolites, Actinobacteriota may affect other bacteria and plants in various ways. The thesis aimed at assessing the connection between the production of secondary metabolites and interactions of actinobacteria in plant and soil environments. In the first part, we assessed how rare actinobacteria from acid soil affect cultivable soil bacteria. We isolated a collection of actinobacteria from acid soil dominated by previously uncultivable lineage and isolated a representative described as a new family Treboniaceae and multiple other potentially novel species of known genera. The genome and metabolome analysis of Trebonia kvetii, sp.nov. gen.nov., demonstrated its ability to produce unusual and potentially novel bioactive metabolites. We designed an experiment where a single strain was co-cultivated with the total soil bacterial community of the same soil in a set up, where a free exchange of diffusible metabolites was allowed between them. Three actinobacteria strains coming from distinct lineages were selected for this interaction. Each strain significantly and specifically affected cultivable bacteria as well as the metabolite pool, part of which was induced...
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Characterization of fungal community in spruce (Picea abies) litter using cultivation and T-RFLP
Kolářová, Zuzana ; Koukol, Ondřej (advisor) ; Brabcová, Vendula (referee)
Fungi have a key role in the decomposition of coniferous litter and affect nutrient cycling in forest ecosystems. Therefore, great emphasis is placed on exploring the diversity of these organisms. The aim of this thesis was to describe fungal diversity in spruce litter and revealed temporal development of this community in a forest regenerating after bark beetle outbreak. Another objective was to compare sites with different length of bark beetle damage. The study area was located in the Bohemian Forest mountain range. Litter bags with spruce needles were placed on the forest soil and several samplings were performed in the course of three years. Diversity of fungi and changes in the fungal community were assessed by two methods: cultivation of needles on 2ř MEA and fingerprinting method T-RFLP. In total 71 fungal species were obtained from needles during a three-year succession by cultivation approach. Using T-RFLP 122 different fragments were generally recorded. The dominant species were Scleroconidioma sphagnicola, Thysanophora penicillioides, Hormonema dematiodes, Ceuthospora pinastri, species of genus Chalara, Trichoderma polysporum, Mycena galopus and unknown species Helotiales sp. 1. Primary saprotrophs occured in the community mainly in first 8 months and then were replaced by basidiomycetes....
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Gene pool of the secondary metabolism in soil bacterial communities
Patrmanová, Tereza ; Kopecký, Jan (advisor) ; Brabcová, Vendula (referee)
The need for new antibiotics and other biologically active compounds is the reason for an increased interest in secondary metabolites of soil bacteria. The phylum Actinobacteria has the dominant position in the soil environment thanks to the potential of producing a broad spectrum of antibiotics and the presence of a number of defense mechanisms preventing the effects of antibiotics. The aim of this thesis was to determine the number of copies of selected secondary metabolic genes in the soils of two sites using designed primers and primers from literature. The design of effective new primers for the detection of selected genes in the soil environment was not achieved in this work, and therefore only primers from literature that had been verified for their specificity were used. In samples taken from soil profiles of two sites, abundances of bacteria, actinobacteria, type II polyketide synthase genes and Erm methyltransferase genes mediating resistance to MLSB antibiotics (macrolides, lincosamides and streptogramins B) were determined by digital PCR. The comparison of the determined copy numbers gave an information about the structure of the bacterial community and the relative abundance of bacteria carrying selected secondary metabolic genes depending on the soil condition changes due to the...
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Assembly, successional development and functioning of microbial communities in deadwood
Bernardová, Natálie ; Brabcová, Vendula (advisor) ; Kopecký, Jan (referee)
Dead wood is one of the most important reservoirs associated with forest ecosystems. In natural forests, its volume is counted in hundreds of m3 ha-1 , whereas it reaches only tens of m3 ha-1 in productive commercial forests. In contrast to soil and plant litter, deadwood is unevenly distributed on the forest floor. The specific physicochemical properties such as high content of recalcitrant polymers, low nitrogen level and impermeability negatively affect the rate of decomposition especially in the initial stages of wood deconstruction. The deadwood decomposition is very slow in comparisons with other substrates, it accumulates and thus it represents the important reservoir of nutrients. This thesis is focused on the structure, development and function of microbial (fungal) community in decomposing deadwood in unmanaged forest. Functional screening of fungi isolated from fruit bodies collected from coarse deadwood was set aside. Physico-chemical properties of deadwood including pH, carbon and nitrogen content and microbial biomass were estimated for four wood decomposition stages and three different tree species. New generation sequencing (Illumina MiSeq platform) was applied for fungal community structure analysis based on ITS2 fragment. Fungal functional screening was based on physico-chemical...
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Gene pool of the secondary metabolism in soil bacterial communities
Patrmanová, Tereza ; Kopecký, Jan (advisor) ; Brabcová, Vendula (referee)
The need for new antibiotics and other biologically active compounds is the reason for an increased interest in secondary metabolites of soil bacteria. The phylum Actinobacteria has the dominant position in the soil environment thanks to the potential of producing a broad spectrum of antibiotics and the presence of a number of defense mechanisms preventing the effects of antibiotics. The aim of this thesis was to determine the number of copies of selected secondary metabolic genes in the soils of two sites using designed primers and primers from literature. The design of effective new primers for the detection of selected genes in the soil environment was not achieved in this work, and therefore only primers from literature that had been verified for their specificity were used. In samples taken from soil profiles of two sites, abundances of bacteria, actinobacteria, type II polyketide synthase genes and Erm methyltransferase genes mediating resistance to MLSB antibiotics (macrolides, lincosamides and streptogramins B) were determined by digital PCR. The comparison of the determined copy numbers gave an information about the structure of the bacterial community and the relative abundance of bacteria carrying selected secondary metabolic genes depending on the soil condition changes due to the...
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Characterization of fungal community in spruce (Picea abies) litter using cultivation and T-RFLP
Kolářová, Zuzana ; Koukol, Ondřej (advisor) ; Brabcová, Vendula (referee)
Fungi have a key role in the decomposition of coniferous litter and affect nutrient cycling in forest ecosystems. Therefore, great emphasis is placed on exploring the diversity of these organisms. The aim of this thesis was to describe fungal diversity in spruce litter and revealed temporal development of this community in a forest regenerating after bark beetle outbreak. Another objective was to compare sites with different length of bark beetle damage. The study area was located in the Bohemian Forest mountain range. Litter bags with spruce needles were placed on the forest soil and several samplings were performed in the course of three years. Diversity of fungi and changes in the fungal community were assessed by two methods: cultivation of needles on 2ř MEA and fingerprinting method T-RFLP. In total 71 fungal species were obtained from needles during a three-year succession by cultivation approach. Using T-RFLP 122 different fragments were generally recorded. The dominant species were Scleroconidioma sphagnicola, Thysanophora penicillioides, Hormonema dematiodes, Ceuthospora pinastri, species of genus Chalara, Trichoderma polysporum, Mycena galopus and unknown species Helotiales sp. 1. Primary saprotrophs occured in the community mainly in first 8 months and then were replaced by basidiomycetes....
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