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Molecular characterization of BLC response to elicitors/pathogens

 

Principal investigator: Maité Victré-Gibouin/ Marie-Laure Follet-Gueye

Participants: Jean-Claude Mollet, Arnaud Lehner, Laurence Menu-Bouaouiche

PhD students: Rim Jaber


Project:

The effects of the microbe-derived molecules called MAMPS, flg22 (flagellin peptide 22) and PGN (peptidoglycan from B. subtilis), that have been recently shown to trigger tissue-specific responses in Arabidopsis roots (Millet et al., 2010) as well as the pathogenic and non-pathogenic bacteria, Pseudomonas syringae DC3000 and Rhizobium alamii, will be assessed on BLC (collaboration with Dr. C. Santaella, CEA / CNRS / University of Marseille). More specifically, we will investigate the time-lapse impact of a treatment with elicitors and pathogen on BLC production, viability and morphology using microscopy and flow cytometry. We will use specific fluorescent probes and confocal microscopy to examine the production of reactive oxygen species (ROS) and wall molecules such as callose or extensin. Further cell wall alterations will be investigated using microscopy, Fourier Transform Infrared (FT-IR) microspectrometry and chemical analysis. Laser microdissection will be used to isolate BLC for cell wall chemical analysis and also for studying defence genes expression (PR1, THI2.1, PDF1.1). To explore further the response of BLC, metabolomic analyses on microdissected cells will be undertaken to assess possible changes in small molecule metabolites produced upon elicitors/pathogen treatment (collaboration with Pr. E. Gontier, University of Picardie, Amiens).