engleski

Reactive oxygen species (ROS) confocal laser scanning imaging in TROL mutants

Photosynthesis is the process in photoautotrophic organisms that converts light energy into chemical energy. The very important part of this process is the transfer of electrons from reduced ferredoxin to NADP+, which is catalyzed by the flavoenzyme Ferredoxin-NADP+ oxydoreductase (FNR) [1]. Binding of the FNR to photosynthetic membranes of many vascular plants is assisted by the thylakoid rhodanase-like protein (TROL) [2]. Within my PhD project, I am examining the influences of the TROL-FNR protein complex on electron transport and ROS scavenging, which indicates a protective role of this complex. I will carry out the detection and visualization of ROS using fluorescent probes (DHE for superoxides, SOSG for singlet oxygen molecules, spy-LHP for peroxides) for staining ROS molecules and confocal laser scanning microscopy technique for detecting them. This experiment will show the presence and propagation of ROS molecules in chloroplasts and whole plant cells. Imaging is performed using a CCD camera. Arabidopsis thaliana (L.) Heynh was used as a model plant ; wild type (WT), TROL KO, ∆RHO mutants were grown under normal humidity and in arid conditions. The main goal of these experiments is to demonstrate how TROL-FNR complex affects electron transport, ROS scavenging, management and distribution of high energy electrons in energetic processes of photosynthesis [3] under normal soil humidity conditions and under abiotic stress conditions, in this case drought, and how deficiency of some components of this complex affects the efficiency of the TROL-FNR pair. The research will contribute to the understanding of the behavior of plants under severe stress conditions

microscopy ; imaging ; ROS ; TROL

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