Confocal laser-scanning microscopy was chosen to observe the colonization and damage caused by the soft rot Pectobacterium atrosepticum and the protection mediated by the biocontrol agent Rhodococcus erythropolis. We developed dual-color reporter strains suited for monitoring quorum-sensing and quorum-quenching activities leading to maceration or biocontrol, respectively. A constitutively expressed cyan or red fluorescent protein served as a cell tag for plant colonization, while an inducible expression reporter system based on the green fluorescent protein gene enabled the simultaneous recording of signaling molecule production, detection, or degradation. The dual-colored pathogen and biocontrol strains were used to coinoculate potato tubers. At cellular quorum, images revealed a strong pectobacterial quorum-sensing activity, especially at the plant cell walls, as well as a concomitant rhodococcal quorum-quenching response, at both the single-cell and microcolony levels. The generated biosensors appear to be promising and complementary tools useful for molecular and cellular studies of bacterial communication and interference.

Institute of Microbiology of the CAS v v i Vídeňská 1083 CZ 14220 Praha 4 Czech Republic

Laboratoire de Microbiologie Signaux et Microenvironnement Normandie Université LMSM 55 rue Saint Germain 27000 Evreux France and Structure Fédérative de Recherche Normandie Végétale 4277

Laboratoire de Microbiologie Signaux et Microenvironnement Normandie Université LMSM 55 rue Saint Germain 27000 Evreux France and Structure Fédérative de Recherche Normandie Végétale 4277 2 Seeds Innovation Protection Research and Environment Route de la petite chaussée 76110 Bretteville du Grand Caux and Rue des Champs Potez 62217 Achicourt France

Seeds Innovation Protection Research and Environment Route de la petite chaussée 76110 Bretteville du Grand Caux and Rue des Champs Potez 62217 Achicourt France 3 French Federation of seed potato growers 43 45 rue de Naples 75008 Paris France

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