Molecular methods for the specific detection of Colletotrichum sansevieriae
DOI:
https://doi.org/10.15517/am.2025.62815Keywords:
RFLP, anthracnose, plant disease, β-tubulin gene, plant pathogen, fungal diagnosticsAbstract
Introducción. La antracnosis de Sansevieria, causada por Colletotrichum sansevieriae, representa un riesgo significativo para el cultivo y la exportación de esta planta ornamental. Los métodos efectivos y rápidos de identificación de este patógeno son cruciales para implementar medidas de control que prevengan su propagación a áreas no infectadas. Objetivo. Implementar y optimizar métodos moleculares para la identificación rápida y confiable de C. sansevieriae. Materiales y métodos. Durante 2016, se analizó un fragmento del gen β-tubulina-2 (β-tub2) de C. sansevieriae aislado de una finca local en Alajuela, Costa Rica. Se implementó PCR-RFLP del fragmento parcial del gen β-tubulina-2 (β-tub2) con la enzima MseI (Tru1I). Además, se aplicaron cebadores específicos para la detección de C. sansevieriae y análisis de PCR-RFLP del fragmento amplificado. Resultados. La digestión produjo de manera consistente un patrón de restricción de dos bandas específico para C. sansevieriae. Los cebadores diseñados amplificaron con éxito un fragmento de 383 pb del β-tub2 de todas las cepas de C. sansevieriae probadas. No se observó amplificación de otras especies de Colletotrichum dentro de los complejos C. gloesporioides y C. acutatum, ni de aislamientos de C. truncatum y Fusarium oxysporum. Además, este sitio de restricción, ubicado dentro del amplicón generado por los cebadores específicos para C. sansevieriae, permitió la validación exitosa de la especie mediante digestión. Conclusiones. Ambos métodos basados en PCR demostraron ser lo suficientemente sensibles como para detectar C. sansevieriae en hojas de Sansevieria infectadas de manera natural y artificial sin necesidad de aislar el patógeno en cultivos puros, lo que hace que el proceso diagnóstico sea más eficiente y accesible.
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