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One of the first reactions of plants under stress is the enhanced production of chemically distinct reactive oxygen species (ROS). A major difficulty in elucidating the biological activity of ROS during stress stems from the fact that not only one but several chemically distinct ROS are generated simultaneously, thus making it very difficult to link a particular stress response to a specific ROS. This problem has been alleviated by using the conditional flu mutant of Arabidopsis that allows to induce the production of only one ROS, singlet oxygen, within plastids in a non-invasive, controlled manner (Fig. 1). In the dark the flu mutant accumulates protochlorophyllide (Pchlide), a potent photosensitizer that upon illumination generates singlet oxygen. Several singlet oxygen-mediated stress responses have been distinguished during re-illumination of the flu mutant. Inactivation of nuclear genes encoding the two closely related plastid proteins Executer1 and Executer2 has been shown to be sufficient to abrogate these stress responses despite the ongoing release of singlet oxygen. By varying the length of the dark period, one can adjust the level of the photosensitizer Pchlide and define conditions that minimize the cytotoxicity of singlet oxygen and either endorse acclimation in flu plants exposed to a very short dark period as one extreme or promote a genetically controlled cell death response in plants shifted for a longer period to the dark as another extreme (Fig. 2). This activity of singlet oxygen assigns a new function to the chloroplast, namely that of a sensor of environmental changes that activates a broad range of stress responses, known to be activated also by abiotic and biotic stressors. Our work is aimed at dissecting the complexity of singlet oxygen signalling and understanding and eventually also modifying the genetic constraints that determine the adaptability of plants to environmental changes.
Figure 1 FLU-dependent control of light-dependent Chl biosynthesis. Inactivation of FLU impedes negative feedback control (A) and leads to the overaccumulation of excess Pchlide in etiolated flu seedlings that upon excitation with blue light emits a strong red fluorescence (B, DD). Upon light exposure Pchlide acts as a photosensitizer and triggers the release of 1O2 that results in the rapid bleaching of seedlings (B, DL). Under continuous light Pchlide is immediately reduced via POR to Chlide and does not reach critical levels that may evoke the production of 1O2 (B, LL).
Figure 2 Schematic diagram of 1O2-dependent signal transduction from the plastid to the nucleus that triggers stress responses ranging from acclimation to cell death. The quality of the response can be modulated by shifting flu plants for various lengths of time to the dark, by activating modulators (e.g. PRL1) or by inactivating EX1 and/or EX2.