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In Vitro Analysis of E3 Ubiquitin Ligase Function
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Ubiquitin E3 ligase activity of

Xue Ouyang1, Jialan Chen1, Zhimao Sun2

  • 1Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.

Frontiers in Microbiology
|June 9, 2023
PubMed
Summary

The E3 ligase activity of Ralstonia solanacearum effector RipAW is not essential for triggering plant immunity, although it is required for cell death induction. This finding uncouples effector-induced cell death from immune responses in plants.

Keywords:
Ralstonia solanacearumRipAWSGT1cell deathplant immunity

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Area of Science:

  • Plant Pathology
  • Bacterial Phytopathogens
  • Plant-Microbe Interactions

Background:

  • Ralstonia solanacearum causes significant crop losses.
  • Type III effectors mediate R. solanacearum-plant interactions.
  • RipAW, a novel E3 ligase effector, induces cell death in Nicotiana benthamiana.

Purpose of the Study:

  • To decipher the role of RipAW's E3 ligase activity in plant immunity.
  • To investigate the relationship between RipAW-induced cell death and immune responses.
  • To identify components required for RipAW-mediated immunity.

Main Methods:

  • Generated and analyzed E3 ligase activity mutants (RipAWC177A) and truncated mutants of RipAW.
  • Assessed cell death induction and plant immune responses in Nicotiana benthamiana.
  • Investigated the requirement of SGT1, EDS1, NRG1, NRC proteins, and the SA pathway.

Main Results:

  • RipAW's E3 ligase activity is dispensable for triggering plant immunity but required for cell death.
  • N-terminus, NEL domain, and C-terminus are necessary but not sufficient for RipAW-induced cell death.
  • All RipAW mutants triggered Effector-Triggered Immunity (ETI) responses.
  • RipAW-triggered immunity requires SGT1 but not EDS1, NRG1, NRC proteins, or the SA pathway.

Conclusions:

  • RipAW's E3 ligase activity can be uncoupled from effector-triggered plant immunity.
  • This study provides insights into the mechanisms of effector-triggered immunity.
  • Findings offer a model for understanding effector functions in plant defense.