Protocol for histological analysis of the interaction Vanilla planifolia - Fusarium oxysporum f. sp. vanillae

Fusarium oxysporum f. sp. vanillae (Fov) is the main phytosanitary problem of Vanilla planifolia, invading root and causing rotting up to 80% plants. The histological process used study morphological characteristics tissues interest published a protocol check Fov-V. planifolia interaction that was not reproducible under our conditions, so aimed adapt procedure allows differential analysis mechanism Fov invasion in V. by confocal microscopy. bioassays consisted analyzing Fov-infected roots. experimental design three replicates evaluated at 12, 24, 36, 48, 60 72 hours post-inoculation. Roots without inoculum were as controls. samples fixed paraformaldehyde 4% sucrose (PBS). Subsequently, dehydrated combination alcohols different concentrations. cuts made with microtome. sections observed Leica microscope 63X objective ap. No. 1.4, plus immersion oil; excitation lines: 405, 488, 532. FAA + kerosene method allowed complete tissue sections. Keywords: Fusariosis, Vanilla, interaction.