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Photosynthetic cell suspension cultures quantitative image data processing
Vlachynská, Alžběta ; Búzová,, Diana (referee) ; Červený,, Jan (advisor)
This work was carried out in collaboration with the Department of Adaptive Biotechnologies, Global Change Research Centre AS CR. It deals with the quantitative analysis of photosynthetic cell cultures. It uses images captured by a confocal fluorescent microscope to the automatic determining the number of cells in the sample. The work consists of a theoretical analysis, which briefly describes fluorescence and confocal microscopy. It also concisely introduces a microscope Leica TCS SP8 X, which I used to scan data. One capture is devoted to the theory of digital image processing. The second part deskribes the development of algorithm for processing 3D data and simplified algorithm for processing 2D data and its program implementations in a programming environment MATLAB R2013b. Grafical user interface is explained in detail. Done measurement are presented at the conclusion. It mentions compiled sample preparation protocol. The results of the program are compared with manual counting. Number of cells per 1 ml are determined by created program in samples of cell cultures Chenopodium rubrum (Cr) and Solanum lycopersicum (To).
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Ochrana rajčat ve skleníkových a polních podmínkách
Mokrá, Kristýna
The main aim of my bachelor thesis was to evaluate the infection of several varieties of edible tomato by pathogens in one locality, in a greenhouse, and under field conditions. The next step was to compare the obtained results with the results of the infection at the nearest observation point of ÚKZÚZ. The attention was paid to the cultivated varieties Zlatava, Blueberry, Start F1 and to the determined pathogens such as Alternaria spp., Fulvia fulva and Phytophthora infestans. The survey was conducted in 2020 in a private garden. Tomatoes were monitored during the growing season according to BBCH phases and parts infected by pathogens were evaluated according to the Methodology of utility value tests of ÚKZÚZ. The infected parts were cultured on a PDA in a laboratory, followed by the determination of pathogens using a laboratory microscope. The results of infection of individual pathogens on the selected variety in a specific environment were evaluated and expressed in a graphical representation.
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Photosynthetic cell suspension cultures quantitative image data processing
Vlachynská, Alžběta ; Búzová,, Diana (referee) ; Červený,, Jan (advisor)
This work was carried out in collaboration with the Department of Adaptive Biotechnologies, Global Change Research Centre AS CR. It deals with the quantitative analysis of photosynthetic cell cultures. It uses images captured by a confocal fluorescent microscope to the automatic determining the number of cells in the sample. The work consists of a theoretical analysis, which briefly describes fluorescence and confocal microscopy. It also concisely introduces a microscope Leica TCS SP8 X, which I used to scan data. One capture is devoted to the theory of digital image processing. The second part deskribes the development of algorithm for processing 3D data and simplified algorithm for processing 2D data and its program implementations in a programming environment MATLAB R2013b. Grafical user interface is explained in detail. Done measurement are presented at the conclusion. It mentions compiled sample preparation protocol. The results of the program are compared with manual counting. Number of cells per 1 ml are determined by created program in samples of cell cultures Chenopodium rubrum (Cr) and Solanum lycopersicum (To).
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Expression of SANT/HTH Myb mRNA, a plant morphogenesis-regulating transcription factor, changes due to viroid infection
LOMNICKÁ, Anna
Potato spindle tuber viroid (PSTVd) belongs to plant-pathogenic, circular, non-coding RNAs. Its propagation is accompanied by (mis)regulation of host genes and induction of pathogenesis symptoms including changes of leaf morphogenesis depending on the strength of viroid variant. We found strong genotype-dependent suppression of tomato morphogenesis-regulating transcription factor SANT/HTHMyb (SlMyb) due to viroid pathogenesis. Its relative mRNA level was found to be significantly decreased in PSTVd-sensitive tomato (cvs Rutgers and Heinz 1706) due to degradation processes, but increased in PSTVd-tolerant (cv. Harzfeuer). In heterologous system of Nicotiana benthamiana, we observed a SlMyb-associated necrotic effect in agroinfiltrated leaf sectors during ectopic overexpression. Leaf sector necroses were accompanied by activation of nucleolytic enzymes but were suppressed by a strongly pathogenic PSTVd variant. Contrary to that, PSTVd's effect was inhibited by the silencing suppressor p19. It was found that in both, Solanum lycopersicum leaves and N. benthamiana leaf sectors, SlMyb mRNA degradation was significantly stronger in viroid-infected tissues. Necroses induction as well as gene silencing experiments using the SANT/HTH-Myb homologues revealed involvement of this Myb in physiological changes like distortions in flower morphogenesis and growth suppression.
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The comparison of biolistic and Agrobacterium mediated transformation of tomato (Solanum lycopersicum L.)
DĚDOUCHOVÁ, Martina
Currently we can do genetic manipulation by means of direct and indirect methods. Indirect methods use strategy of gram-negative soil bacteria Agrobacterium tumefaciens, which introduces part of its own genes carried by the section of plasmid Ti called T-DNA into the genome of infected plants. It is difficult to reach the transformation of monocotyledonous plants using this method. Application of direct methods of transformation offers the possibility for transformation of monocotyledonous plants. One of the most effective methods of direct transformation is biolistic transformation. This diploma thesis dealt with the comparison of efficacy of biolistic and A. tumefaciens mediated transformation of tomato (Solanum lycopersicum L.).
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