Genetic analysis of endosomal compartments using high throughput imaging

• Applicant: Dr. Martina Beck
• Subject Area: Plant Cell and Developmental Biology
• Term: from 2011 to 2014
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 198122488

Membrane compartmentalization and trafficking within and between cells is considered an essential cellular property of higher eukaryotes. The formation of endomembrane compartments, plasma membrane (PM) and cell walls relies on vesicular trafficking which involves the formation, transport and fusion of vesicles with target membranes. Thus, plant vesicular trafficking is necessary for development, signal transduction and responses to biotic and abiotic stresses. For example, dynamic processes at the plasma membrane are important for immune reactions upon pathogenic microorganisms. Although numerous membrane trafficking components have been identified in plants, the dynamics of the underlying processes in intact cells, the coordination of cellular subfunctions between different endomembrane compartments, as well as their tissue- and cell type-specific differentiation are still poorly understood. The straightforward approach to analyze the organization of the endosomal system within plants is to identify the intracellular compartments that are involved in this trafficking route. A high-throughput imaging method suitable for the quantitative detection of membrane compartments at subcellular resolution in intact epidermal tissue was established and applied in a genetic screen of a transgenic Arabidopsis line expressing GFP-2xFYVE as a marker for late endosomal/Multi vesicular bodies (MVB) compartments. This approach allowed the identification of mutants with altered endosomal levels, reflecting in either high level of endosomes or reduced level of endosomes. Several fel (FYVE-endosome levels) mutants could be confirmed. The proposed project deals with the mapping and characterization of the responsible genes which will help to identify new candidates possibly involved in induced endo- and exocytotic events.

• DFG Programme: Research Fellowships
• International Connection: United Kingdom
• Host: Professorin Dr. Silke Robatzek