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Controlled Activation of NLR-Mediated Immunity in Roots Using a Copper-Inducible Expression System.

Hiroshi Yoshida1, Kodai Honda2, Saki Matsuoka1

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Summary

Researchers developed a copper-inducible system for tomato hairy roots to study plant immunity. This tool precisely activates immune signaling, revealing nucleotide-binding leucine-rich repeat receptor (NLR) dynamics and cell death in roots.

Keywords:
NLRcopper-inducible expression systemhairy root transformationhypersensitive responseplant root immunitysubcellular dynamics

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Area of Science:

  • Plant Biology
  • Molecular Biology
  • Immunology

Background:

  • Plant nucleotide-binding leucine-rich repeat receptors (NLRs) are crucial for immunity in shoots and roots.
  • The specific functions of NLRs in root immunity are not well understood.

Purpose of the Study:

  • To establish a controllable gene expression system for studying plant immune signaling in tomato roots.
  • To investigate the roles and dynamics of NLRs in root immune responses.

Main Methods:

  • Developed a copper-inducible gene expression system in tomato hairy roots using CUP2 transcription factor and a CBS4-miniDFR promoter.
  • Integrated a fluorescent marker for easy identification of transgenic root cells.
  • Induced autoactive NLR variants (NRC4DV-EGFP, Gpa2NB-EGFP) to trigger immune responses.

Main Results:

  • The system allowed precise, on-demand activation of immune signaling with minimal basal expression.
  • Induced NLR variants triggered reactive oxygen species (ROS) accumulation and hypersensitive cell death in roots.
  • Confocal imaging showed distinct subcellular dynamics of NLRs in root cells upon induction.

Conclusions:

  • The developed system is a robust and versatile tool for investigating root immune signaling.
  • It enables controlled study of NLR dynamics and function in plant roots.
  • Findings are consistent with NLR behavior observed in plant leaf cells.