The Sources Of ROS Generated In The Interaction Between Arabidopsis And Pseudomonas Syringae Pv. Tomato DC3000 Investigated By RoGFP1

At the preliminary stage of the interaction between plant and the pathogens, an oxidative burst happens, generating amounts of reactive oxygen species. It is believed popularly that these reactive oxygen species are from the NADPH oxidase compounds associating with plasma membrane, the oxidase system in the cell wall and the pathogen in the interaction system. In this thesis, a reduction-oxidation sensitive green fluorescence protein is used to examine the redox potential changes in cytoplasma and mitochondria in situ, in real time, and without damage to the examined materials when Arabidopsis interacts with Pseudomonas syringae pv. tomato DC3000 in order to speculate the concentration changes of reactive oxygen species. In this thesis, the first part acts to introduce the current status and progression of plant-pathogen interaction research and the development and application of redox sensitive GFP. The second part is to confirm that the redox sensitive GFP was sensitive to the environmental redox potential changes when expressed in our Arabidopsis materials, suggesting that the redox status in Arabidopsis cells could be examined using this kind of GFP. The third part focuses on the construction of the model of infection when DC3000 is inoculated into Arabidopsis, which could be accordingly applied to research redox potential changes of Arabidopsis cells occurring in the Arabidopsis-DC3000 interaction system. The fourth part is about the respiration during the Arabidopsis-DC3000 interaction, which showed an increase in leaves inoculated and leaves with Systemic Acquired Resistance, suggesting that there was probably an increase in amount of reactive oxygen species in Arabidopsis cells. And the fifth part demonstrates that the oxidation status had increased in mitochondria of leaves inoculated and leaves with Systemic Acquired Resistance, implying the initiation of reactive oxygen species probably also had been enhanced, when the redox potentials were examined in a real-time, in situ and damage-free way. And then the sixth part concludes that, after the inoculation of DC3000, the Arabidopsis mitochondria of leaves inoculated and leaves with Systemic Acquired Resistance may generate much reactive oxygen species, and the inoculation of H2O may also cause an oxidative burst to some extend in the region inoculated and the region far away. To summarize, mitochondria of Arabidopsis is a likely source of reactive oxygen species in the interaction system of Arabidopsis-DC3000.