We describe a fluorescence imaging method to visualize the dynamics of the central vacuole in rice cells during invasion by the blast fungus Magnaporthe oryzae. This method utilizes the combination of confocal microscopy, rice sheath cells (optically transparent), fluorescently tagged M. oryzae (red fluorescence), and fluorescein diacetate staining (green fluorescence; visualizing vacuole dynamics). Using this method, we demonstrate that the vacuole undergoes progressive shrinkage and collapse during M. oryzae infection.
Keywords: Abiotic and biotic stress; Confocal imaging; Fluorescein diacetate; Magnaporthe oryzae; Oryza sativa; Plant cell death; Plant–pathogen interactions; Rice blast; Vacuolar dynamics; Vacuolar staining.