Příprava a testování bioluminiscenčních sensorů pro cytokininy v Arabidopsis thaliana

Novotná, Lenka

Advances in microscopy and the sensitivity of novel biosensors have enabled the development of new methods for monitoring phytohormones with low invasiveness and relatively high resolution, providing greater insight into phytohormone function in plants. Genetically encoded indirect biosensors for cytokinins, one of the main plant hormones, are widely used for the tracking of dynamic changes in their levels during plant development and growth. The cytokinin-sensitive synthetic promoters used as cytokinin biosensors are being continuously improved, providing better insight into cytokinin function. However, the reporter systems used for the visualizations are either based on the enzymatic reaction producing colour or on the usage of fluorescence proteins. Luminescence-based biosensors are often used to visualize signaling outputs of plant hormones, but they are based on exogenous addition of luciferin substrate. Therefore, in this thesis, I have introduced a new technique for the generation of cytokinin signaling output sensors based on autonomous bioluminescence. This autonomous bioluminescence represents a non-invasive technique that depends on the expression of the whole autonomous bioluminescence system from fungi Neonothopanus nambi, in particular, the caffeic acid cycle with the fungal...

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