A Phytophthora effector protein promotes symplastic cell-to-cell trafficking by physical interaction with plasmodesmata-localised callose synthases.

Pathogen effectors act as disease promoting factors that target specific host proteins with roles in plant immunity. Here, we investigated the function of the RxLR3 effector of the plant-pathogen Phytophthora brassicae. Arabidopsis plants expressing a FLAG-RxLR3 fusion protein were used for co-immunoprecipitation followed by liquid chromatography-tandem mass spectrometry to identify host targets of RxLR3. Fluorescently labelled fusion proteins were used for analysis of subcellular localisation and function of RxLR3. Three closely related members of the callose synthase family, CalS1, CalS2 and CalS3, were identified as targets of RxLR3. RxLR3 co-localised with the plasmodesmal marker protein PDLP5 (PLASMODESMATA-LOCALISED PROTEIN 5) and with plasmodesmata-associated deposits of the ß-1,3-glucan polymer callose. In line with a function as an inhibitor of plasmodesmal callose synthases (CalS) enzymes, callose depositions were reduced and cell-to-cell trafficking was promoted in the presence of RxLR3. Plasmodesmal callose deposition in response to infection was compared with wild-type suppressed in RxLR3-expressing Arabidopsis lines. Our results implied a virulence function of the RxLR3 effector as a positive regulator of plasmodesmata transport and provided evidence for competition between P. brassicae and Arabidopsis for control of cell-to-cell trafficking.