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Specimen Preparation and Observations of Magnaporthe oryzae Appressorial Cells Under Electron Microscopy

Raquel O Rocha1, Richard A Wilson2

  • 1Department of Plant Pathology, University of Nebraska-Lincoln, Lincoln, NE, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 8, 2021
PubMed

Insights

This study presents two cryo-electron microscopy preparation methods for visualizing Magnaporthe oryzae appressoria on hydrophobic surfaces. These techniques overcome common sample fixation challenges for improved cell ultrastructure analysis.

Area of Science:

  • Plant pathology
  • Microscopy techniques
  • Cell biology

Background:

  • Electron microscopy (EM) is crucial for cellular morphology and ultrastructure analysis.
  • Cryo sample fixation presents significant challenges, hindering widespread EM adoption.
  • Understanding fungal-pathogen interactions requires advanced imaging methods.

Purpose of the Study:

  • To describe two novel EM preparation protocols for studying Magnaporthe oryzae appressoria.
  • To address cryo sample fixation challenges in EM.
  • To enable detailed ultrastructural analysis of appressoria on artificial hydrophobic surfaces.

Main Methods:

  • Development and application of two distinct EM sample preparation techniques.
  • Utilizing artificial hydrophobic surfaces for appressoria cultivation.
  • Employing electron microscopy for ultrastructural examination.

Main Results:

  • Successful visualization of Magnaporthe oryzae appressoria ultrastructure.
  • Demonstration of effective sample preparation overcoming fixation issues.
  • Acquisition of high-resolution images of appressoria on hydrophobic substrates.

Conclusions:

  • The described EM protocols provide viable alternatives for studying fungal appressoria.
  • These methods facilitate detailed ultrastructural investigations of plant-pathogen interactions.
  • Overcoming cryo fixation challenges enhances the utility of EM in mycology.
Keywords:
AppressoriumElectron microscopyImagingMagnaporthe oryzaeSEMSample preparationSurfacesTEM

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