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RETRaIN Report Summary

Project ID: 310901
Funded under: FP7-IDEAS-ERC
Country: United Kingdom

Final Report Summary - RETRAIN (REwiring the nucleus TowaRds plant ImmuNity)

Pathogens greatly hamper food production but only a few microbes can form intimate associations with plants. Plant Pattern Recognition Receptors perceive Microbe or Pathogen Associated Molecular Patterns (PAMPs) and activate Pattern Triggered Immunity. Pathogens perturb these cellular processes to suppress immunity. Indeed, functional genomics and biochemical studies have unveiled specialized secreted pathogen proteins (effectors) that trigger susceptibility. This has led to the view that plant-pathogen interactions constitute dynamic interplay between host defenses and specialized pathogen machinery that aim to subvert immunity.

With this proposal, we aimed to unravel the molecular mechanisms that underpin infection outcomes in the nucleus. Quantitative proteomics, gene expression analyses, and localization studies were used to (i) detect pathogen effectors in the host nucleus in situ and (ii) identify changes in the nuclear proteome during infection. Having identified the changes and effectors associated with infection, we then employed Yeast two Hybrid and immunoprecipitation experiments to identify effector host targets. Screening of over 25 effectors helped identify a range of candidate host targets, some of which were validated in vivo and appear to contribute to immunity. Amongst the targets identified, we characterized the activity of Importin proteins towards their substrates. Immunoprecipitation experiments in N. benthamiana plants infected with P. capsici, helped define changes in specificity towards importin cargo proteins, which may underpin quantitative changes in the nuclear proteome during infection.

Towards a proof of concept, we sought to generate synthetic LRRs that could be used in an immune signaling context to enhance immunity. Y2H based screening of pool-based screening of a small sLRR library helped identify a set of 5 candidate sLRRs, able to bind a host factor in yeast. Validation of these interactions in vivo will allow work aimed at testing this approach in plants.

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United Kingdom
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