Protocol for histological analysis of the interaction Vanilla planifolia - Fusarium oxysporum f. sp. vanillae
Por:
Alejandro Q.V., Mónica R.V., Elizabeth V.L.O., Andrés R.-F., Mauricio L.-R.
Publicada:
1 ene 2023
Resumen:
Fusarium oxysporum f. sp. vanillae (Fov) is the main phytosanitary problem of Vanilla planifolia, invading the root and causing the rotting of up to 80% of the plants. The histological process is used to study the morphological characteristics of the tissues of interest and published a protocol to check Fov-V. planifolia interaction that was not reproducible under our conditions, so the study aimed to adapt a procedure that allows differential analysis of the mechanism of Fov invasion in V. planifolia by confocal microscopy. The bioassays consisted of analyzing Fov-infected roots. The experimental design consisted of three replicates evaluated at 12, 24, 36, 48, 60 and 72 hours post-inoculation. Roots without inoculum were used as controls. The samples were fixed in paraformaldehyde and 4% sucrose (PBS). Subsequently, the tissues were dehydrated by a combination of alcohols at different concentrations. The cuts were made with a microtome. The sections were observed under a Leica confocal microscope with a 63X objective with ap. No. 1.4, plus immersion oil; excitation lines: 405, 488, 532. The FAA + kerosene method was the procedure that allowed complete tissue sections. © 2022 by the authors.
Filiaciones:
Alejandro Q.V.:
Laboratorio de Genética e Interacciones Planta Microorganismos, Facultad de Ciencias Agrícolas, Universidad Veracruzana, Mexico
Mónica R.V.:
Facultad de Ciencias, Universidad Nacional Autónoma de México (UNAM), Mexico
Elizabeth V.L.O.:
Instituto de Ecología A.C. El Haya, Veracruz, Xalapa, Mexico
Andrés R.-F.:
Laboratorio de Genética e Interacciones Planta Microorganismos, Facultad de Ciencias Agrícolas, Universidad Veracruzana, Mexico
Mauricio L.-R.:
Laboratorio de Genética e Interacciones Planta Microorganismos, Facultad de Ciencias Agrícolas, Universidad Veracruzana, Mexico
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