An Arabidopsis and tomato mesophyll protoplast system for fast identification of early MAMP-triggered immunity- suppressing effectors

Malou Fraiture, Xiangzi Zheng, Frédéric Brunner

Research output: Contribution to journalArticleResearchpeer-review

18 Citations (Scopus)

Abstract

Transient expression in plant mesophyll protoplasts allows rapid characterisation of gene functions in vivo in a simplifi ed and synchronized manner without bias due to the use of bacteria-based gene or protein delivery systems. It offers the possibility to test whether microbial effectors can subvert early events of plant immune signaling that are activated upon recognition of Microbe-Associated Molecular Patterns (MAMPs), the so-called MAMP-triggered immunity (MTI). Here, we describe the isolation and transfection with effector genes of Arabidopsis thaliana and Solanum lycopersicum mesophyll protoplasts, the use of a non-invasive luciferase reporter assay and a simple method to detect activated Mitogen-Activated Protein Kinases (MAPKs) to identify and study, in a medium-throughput manner, new effectors suppressing early signal transduction events of MTI.

Original languageEnglish
Pages (from-to)213-230
Number of pages18
JournalMethods in Molecular Biology
Volume1127
DOIs
Publication statusPublished - 2014

Keywords

  • Arabidopsis thaliana
  • Effector
  • MAPK
  • Mesophyll protoplast
  • MTI suppression
  • Reporter assay
  • Solanum lycopersicum
  • Tomato
  • Transient expression

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