RESISTOME · Resolving Structural Interactions Shaping Plant Immunity

Plant diseases destroy crops such as wheat, tomato, and corn and cause losses exceeding €180 billion each year, a threat that climate change makes worse by speeding pathogen evolution and resistance breakdown. The RESISTOME project addresses this threat by mapping the physical contacts between pathogen proteins and plant proteins that trigger or block immunity. Many of these contacts are brief and rare and occur inside intact plant tissue, so they escape conventional methods. To catch them, RESISTOME will develop in vivo cross linking methods adapted for plant cells, a chemical approach that fixes pairs of proteins that touch each other so they can be identified later. These cross links will be recovered and identified by mass spectrometry after testing conditions in both isolated plant cells and whole tissues. At the same time the project will modernize cross linking mass spectrometry by using a fast scanning Orbitrap Astral instrument with narrow window data independent acquisition to increase sensitivity and sampling, allowing detection of low abundance and transient interactions across the proteome. Applied to the model plant Arabidopsis thaliana and to tomato, this workflow will be used during controlled pathogen challenges and time resolved experiments to rebuild the sequence of molecular events that follow infection. Cross link distance measurements will be combined with structural predictions to produce three dimensional models of immune receptor complexes and of how pathogen effectors target susceptibility proteins. The resulting proteome wide structural atlas of plant immune interactions will identify mechanistic targets and assays that can guide breeding and engineering for durable resistance. The MSCA fellowship will support the interdisciplinary training required to integrate proteomics, structural modeling, and plant biology needed to translate methods from model plants to crops.