Extracción de proteínas secretadas por el fitopatógeno Macrophomina phaseolina: Selección de un método eficiente que incluye estímulo con tejido de su hospedero

Autores/as

  • Andrea Benegas Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Asunción
  • Alberto A. Cubilla-Ríos Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Asunción
  • M. Eugenia Flores-Giubi Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Asunción
  • Javier E. Barúa 1Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Asunción
  • Ma. Cristina Romero-Rodríguez Universidad Nacional de Asunción

DOI:

https://doi.org/10.17268/sci.agropecu.2022.017

Palabras clave:

podredumbre carbonosa de la soja, hongo necrotrófico, extracción de proteínas, perfil proteico, SDS-PAGE

Resumen

El hongo necrotrófico, mundialmente distribuido, Macrophomina phaseolina es el agente causal de enfermedades de cultivos de importancia económica como la podredumbre carbonosa de la soja. Este hongo secreta una amplia variedad de proteínas y metabolitos que le permiten invadir la planta e iniciar el proceso de infección. La contribución de las proteínas con actividad hidrolítica secretadas por este hongo en el proceso de infección ha sido ampliamente estudiada, las proteínas sin actividad enzimática también podrían poseer un papel importante en este proceso. El análisis de proteínas totales permitiría ampliar el conocimiento sobre este patógeno, y establecer estrategias más eficientes para su control. El objetivo del presente trabajo fue evaluar tres métodos de extracción de proteínas secretadas por M. phaseolina. El hongo fue crecido en medio líquido PDB y Czapek-Dox (CZP) con y sin suplemento de hojas de soja. Las proteínas fueron extraídas del filtrado liofilizado del medio PDB utilizando tres métodos de extracción, y analizadas mediante SDS-PAGE. Se seleccionó el método de precipitación de proteínas con ácido tricloroacético en acetona ya que mostró una mejor resolución del perfil proteico. El filtrado de M. phaseolina crecido en PDB suplementado con hojas de soja (MpPDBs) presentó un mayor rendimiento de extracción de proteínas secretadas entre todas las condiciones evaluadas. Los perfiles proteicos del medio PDB con y sin suplemento, presentaron siete bandas diferenciales, una de ellas específica, detectada en MpPDBs. Estos resultados constituyen una base para estudios sobre el papel de proteínas secretadas por el hongo en el proceso de infección.

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Publicado

2022-07-05

Cómo citar

Benegas, A., Cubilla-Ríos, A. A., Flores-Giubi, M. E. ., Barúa, J. E., & Romero-Rodríguez, M. C. (2022). Extracción de proteínas secretadas por el fitopatógeno Macrophomina phaseolina: Selección de un método eficiente que incluye estímulo con tejido de su hospedero. Scientia Agropecuaria, 13(2), 185-192. https://doi.org/10.17268/sci.agropecu.2022.017

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