The first part of the thesis is devoted to the study of an epigenetic regulation of genes in unicellular green microalgae. The influence of environment on DNA methylation level, histones modifications, chromatin structure and RNA interferences are described as well as their effect on metabolome. The work focuses more closely on the effect of the demethylation reagent 5-azacytidine and CdCl2 on the amount of 5-mC and metabolites of the methionine cycle in Chlamydomonas reinhardtii and Scenedesmus quadricauda in the context of the production of secondary metabolites. The second part of the thesis is focused on genetic engineering. Specifically, the CRISPR/Cas9 method was used as RNA-directed gene editing in the model organism, C. reinhardtii. An optimization of the cpFTSY chloroplast gene was performed, demonstrating phenotypic change in mutated colonies. Furthermore, CRISPR/Cas9 was used to edit the adiposuppressor gene WDTC1. For both genes we obtained mutants whose DNA was repaired using both NHEJ and HDR. Using the fluorescence NileRed analysis, two ?WDTC1 colonies demonstrated an increased TAG content. Epigenetic mechanisms should be studied more to understand the adaptation of microalgae to stress and the environment, providing valuable information for knowledge of metabolic pathways, transcription factors. This information can be used for controlled gene editing, resulting in increased amounts of product for the same or better biomass growth.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:426235 |
| Date | January 2019 |
| Creators | Bačová, Romana |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/doctoralThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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