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 language | English |
|---|---|
| Pages (from-to) | 213-230 |
| Number of pages | 18 |
| Journal | Methods in Molecular Biology |
| Volume | 1127 |
| DOIs | |
| Publication status | Published - 2014 |
Keywords
- Arabidopsis thaliana
- Effector
- MAPK
- MTI suppression
- Mesophyll protoplast
- Reporter assay
- Solanum lycopersicum
- Tomato
- Transient expression