Visualizing the movement of Magnaporthe oryzae effector proteins in rice cells during infection

Abstract

During biotrophy, filamentous pathogens such as the rice blast fungus Magnaporthe oryzae deliver effector proteins into live host cells to facilitate colonization. We describe three complementary assays for visualizing M. oryzae effector translocation into the rice cytoplasm and cell-to-cell movement during infection. Our assays make use of live-cell confocal imaging of optically clear rice sheath cells infected with transgenic strains of M. oryzae that express the fluorescent protein-tagged effector known as PWL2. We highlight several important considerations for the analysis of effector translocation and movement dynamics during infection of host plants.