Analysis of the phosphoproteome of Arabidopsis after pathogen infection: different responses after virulent and aviurlent Pseudomonas syringae infection?

• Applicant: Dr. Florian Kaffarnik
• Subject Area: Organismic Interactions, Chemical Ecology and Microbiomes of Plant Systems
• Term: from 2004 to 2009
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 5430796

Phosphorylation is an integral part of posttranscriptional modification during signal transduction in plants after pathogen infection. In the requested research project I want to examine changes in global phosphorylation patterns in Arabidopsis during plant-pathogen interactions using a system consisting of Arabidopsis cell suspension cultures and Pseudomonas syringae pv. tomato DC3000. Direct comparison of the phosphoproteome of plant cells infected with avirulent or virulent P. syringae allows the identification of specific components involved in R gene mediated resistance. I will use liquid chromatography and mass spectroscopy for analyses together with a refined method for immobilised ion metal affinity chromatography to enrich phosphopeptides. With this recent improvement in the technology developed in the host laboratory from Dr. Scott Peck (Sainsbury Laboratory, Norwich, UK), it is possible to detect most phosphorylated proteins from a cell extract. In this way identified candidates will be further analysed with T-DNA knock-out lines in Arabidopsis and virus-induced gene silencing (VIGS) in Nicotiana benthamiana. With this global approach I hope to find new components involved in R gene mediated resistance and to make a contribution for understanding complex signaling networks. This should provide the opportunity to gain new insights into plant-pathogen communication.

• DFG Programme: Research Fellowships
• International Connection: United Kingdom
• Cooperation Partner: Professor Dr. Scott Peck