High Light-induced Sign Change of Gravitaxis in the Flagellate Euglena gracilis is Mediated by Reactive Oxygen Species

Euglena gracilis responds to abiotic stress factors (high light, salinity, heavy metals) with a sign change of its gravitactic behavior. This phenomenon is oxygen dependent and can be suppressed by the application of the reductant dithionite. It is not mediated by the photoreceptor since also blind mutants change their movement behavior upon high light exposure. It is also not mediated by the chloroplasts since the gravitactic sign change was also found in white, chloroplast-free mutants. The NO radical donor SNAP and the NO cleaver carboxy-PTIO had no obvious effects on gravitaxis or gravitactic sign change, respectively, indicating that NO radicals are not likely involved in gravitactic sign change. Gravitactic sign change was suppressed when oxygen was removed by flushing the cell suspension with nitrogen. Also, the addition of the radical scavengers Trolox, ascorbic acid or potassium cyanide abolished or reduced gravitactic sign change. Quantification of reactive oxygen species (ROS) in the cells indicated that these treatments reduced the evolution of ROS. Furthermore, addition of hydrogen peroxide induced gravitactic sign change in the absence of external stress factors. These results indicate that gravitactic sign change is triggered by ROS (most likely hydrogen peroxide) which are probably produced by cytochrome-c-oxidase in the mitochondria. The clear responses of Euglena to abiotic stress factors suggest that these cells are probably interesting model systems in the study of stress signaling.