Engineering synthetic sensors and responses in plant to uncover physiological and stress-related responses

The main goal of this Ph.D. project is to expand the toolbox of genetically encoded biosensors in plant, and we tried to achieve this objective in different ways. To quantify the presence of molecules possibly involved in the regulation of the hypoxic response in plants, in particular at the level of the shoot apical meristem (SAM), we designed and tested either new or already characterized sensors for hydrogen sulfide, ethanol, pyruvate, cytosolic MgATP2- concentration, and NADH/NAD+ ratio. In parallel, we performed screenings of an Arabidopsis cDNA Over-eXpressing transgenic line seed library, and an ANAC017 overexpressor line under waterlogging stress, to search for mutants with a different tolerance to the stress that could be studied to gain new insights about the hypoxic response. At the same time, we worked on the design and characterization of a blue-green light-driven switch exploiting the OCP2 protein from Cyanobacteria, to engineer an orthogonal response to blue-green light in higher plants, which also required the implementation of the carotenoid biosynthetic pathway of Arabidopsis thaliana for the production of specific keto-carotenoids.