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Super-resolution imaging of plasmodesmata using three-dimensional structured illumination microscopy.

Jessica Fitzgibbon1, Karen Bell, Emma King

  • 1Institute of Molecular Plant Sciences, University of Edinburgh, Edinburgh EH9 3JR, United Kingdom.

Plant Physiology
|May 29, 2010
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Summary

Three-dimensional structured illumination microscopy (3D-SIM) visualized plant cell structures with unprecedented detail. This super-resolution technique revealed how viral movement protein (MP) interacts with plasmodesmata (PD) and the sieve element reticulum (SER) in tobacco plants.

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Area of Science:

  • Plant cell biology
  • Microscopy
  • Virology

Background:

  • Plasmodesmata (PD) are crucial for intercellular transport in plants.
  • Viral movement proteins (MP) facilitate virus spread by interacting with PD.
  • Understanding MP dynamics within PD and related cellular structures is key to controlling viral infections.

Purpose of the Study:

  • To visualize the ultrastructure of plasmodesmata (PD) and associated viral movement protein (MP) in Nicotiana tabacum using super-resolution microscopy.
  • To investigate the localization and dynamics of MP within PD, sieve elements (SEs), companion cells (CCs), and the sieve element reticulum (SER).

Main Methods:

  • Three-dimensional structured illumination microscopy (3D-SIM) was employed for subdiffraction imaging.
  • Green fluorescent protein (GFP)-tagged viral movement protein (MP) was used to track protein localization.
  • Fluorescence recovery after photobleaching (FRAP) was performed to assess MP mobility.

Main Results:

  • 3D-SIM achieved twice the resolution of confocal microscopy, resolving individual PD components (neck and central cavities).
  • MP-GFP filaments were observed extending from PD into the sieve element reticulum (SER) within phloem cells.
  • The SER interconnects pore-PD at the SE-CC interface and contains fine endoplasmic reticulum threads.
  • MP associated with the SER in SEs showed limited mobility, suggesting sequestration.

Conclusions:

  • 3D-SIM provides critical insights into the subdiffraction organization of plant cellular structures, particularly PD.
  • Viral movement protein (MP) localization and restricted mobility within the sieve element reticulum (SER) suggest a role in regulating intercellular transport.
  • This study highlights the potential of 3D-SIM for advancing plant cell biology and virology research by bridging the gap between confocal and electron microscopy.