Transient expression in Arabidopsis leaf mesophyll protoplast system for cell-based functional analysis of MAPK cascades signaling

Mitogen-Activated Protein Kinase (MAPK) cascade is one of the main signaling components mediating abiotic and biotic stress and hormone information in plants. Plant MAPK study has been impeded with a genetic approach using a long-term phenotypic analysis in spite of the transient nature of the protein kinase signaling. Arabidopsis leaf mesophyll protoplasts provide a versatile resource for diverse cell-based assays to acquire immediate molecular and biochemical responses with transient expression of MAPK cascade components of interests. Thus, it is an attractive tool for a high-throughput functional analysis of Arabidopsis MAPK cascade signaling. However, transient expression in Arabidopsis mesophyll protoplast (TEAMP) system requires mastered skills for protoplast preparation and handling to achieve steady and stable data. Here, we have described two analytical methods for MAPK cascade signaling using TEAMP system.