Plant MAP Kinases pp 13-25
Quantification of Stress-Induced Mitogen-Activated Protein Kinase Expressional Dynamic Using Reverse Transcription Quantitative Real-Time PCR
Although it is generally accepted that signal transduction in plant mitogen-activated protein kinase signaling cascades is regulated via rapid posttranslational modifications, there are also several compelling examples of swift stress induced transcriptional activation of plant MAP kinase genes. A possible function of these fast and transient events is to compensate for protein losses caused by degradation of phosphorylated MAP kinases within stimulated pathways. Nevertheless, there is still need for additional evidence to precisely describe the regulatory role of plant MAP kinase transcriptional dynamics, especially in the context of whole stress stimulated pathways including also other signaling molecules and transcription factors. During the last two decades a reverse transcription quantitative real-time PCR became a golden choice for the accurate and fast quantification of the gene expression and gene expression dynamic. In here, we provide a robust, cost-effective SYBR Green-based RT-qPCR protocol that is suitable for the quantification of stress induced plant MAP kinase transcriptional dynamics in various plant species.
Key wordsPlant mitogen-activated protein kinase Abiotic stress Biotic stress Tri reagent Reverse transcription quantitative real-time PCR SYBR green
- 2.Mizoguchi T, Irie K, Hirayama T, Hayashida N, Yamaguchi-Shinozaki K, Matsumoto K, Shinozaki K (1996) A gene encoding a mitogen-activated protein kinase kinase kinase is induced simultaneously with genes for a mitogen-activated protein kinase and an S6 ribosomal protein kinase by touch, cold and water stress in Arabidopsis thaliana. Proc Natl Acad Sci U S A 93:765–769PubMedCentralPubMedCrossRefGoogle Scholar
- 18.Fey A, Eichler S, Flavier S, Christen R, Hofle MG, Guzman CA (2004) Establishment of a real-time PCR-based approach for accurate quantification of bacterial RNA targets in water, using Salmonella as a model organism. Appl Environ Microb 70:3618–3623Google Scholar