Classical biochemical in vitro methods for studying plant biotic interactions are time-consuming, invasive and they provide insight only into the single state of the cell. Therefore, the development of biosensors which enable non-invasive monitoring of the plant immune response in vivo with high spatiotemporal resolution are crucial. The purpose of this master thesis was the optimisation of potato sensor plants constuction for the studies of immune signaling response. Using Plant X-tender technology, we prepared a cytokinine biosensor for following intercellular cytokinins concentration and we tested its function in tobacco plants by transient transformation. With a stable transformation, we prepared transgenic potato plants of Rywal and Désirée genotypes containing Jas9-VENUS biosensor and transgenic potato plants of NahG-Rywal genotype containing biosensor of redox state. Once we obtained transgenic lines with pt-roGFP2 redox state sensor, we selected those with the most intensive fluorescence. In addition, we developed a protocols for the redox state detection in chloroplasts using confocal microscopy, as well as confocal image analysis. We tested the fuction of the sensor with a reductant and oxidant. We determined the redox state of chloroplasts with high spatiotemporal resolution in PVY-inoculated plants. We showed that after the infection, the redox state in chloroplasts is dependent on the distance from the lesions. Chloroplasts next to the lesions are in oxidized state, which might reduces the intercellular transport and the possibility of spreading the virus outside of the infected area. Chloroplasts that are further from the lesions are in reduced state with a normal intercellular transport.
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