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Molecular biology of soil fungi participating in litter decomposition in forest ecosystems
Voříšková, Jana
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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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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Molecular biology of soil fungi participating in litter decomposition in forest ecosystems
Voříšková, Jana
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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The use of comet assay for the estimation of DNA damage during the anticancer chemotherapy.
Voříšková, Jana ; Kovařík, Miroslav (advisor) ; Pávek, Petr (referee)
We have monitored the DNA damage induced by tumor chemotherapy. The combination of platin derivate (cis-platine or carboplatin) and another chemotherapy agent (gemcitabin, paclitaxel or vinorelbin) was applied to patients with lung carcinoma. The whole chemotherapy includes 4 cycles. We have monitored the DNA damage in peripheral blood lymphocytes through the first and the third cycle of chemotherapy and 3 weeks after the last cycle of chemotherapy. For detection we have used comet assay which seems to be accurate and reliable method. Lymphocytes were isolated from whole blood through the medium LSM lymfocyte and then were processed by the alkaline version of the single cell gel electrophoresis. We have analysed single strand breaks (SSB) and interstrand cross links (ICL). For the measurement of ICL isolated lymphocytes were incubated in the presence of 200 µM of styrene oxide (SO) to degrade the DNA by inducing strand breaks. Incubation of peripheral blood lymphocytes with 200 µM SO leads to the formation of comets. If ICLs are present in the DNA, the alkaline unwinding of DNA is blocked and the % of DNA in tail is reduced. Comets were stained with ethidium bromide and then analyzed by the software LUCIA Comet Assay. The damage of DNA caused by SSB before the chemotherapy was relatively low and...
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