Engineering new-to-nature immune sensors for novel recognition of Magnaporthe oryzae

Phytopathogenic fungi significantly impact global food security, causing up to 40% of annual crop yield losses worldwide. Alone, the blast fungus Magnaporthe oryzae has a global economic impact of $66 billion per annum. Phytopathogens deploy intracellular virulence proteins known as effectors that promote pathogenesis through interaction with host plant proteins. Resistance against phytopathogens can occur through recognition of these effectors by intracellular immune receptors known as NLRs. Some NLRs recognise effectors through direct interaction between effectors and integrated domains (IDs), which have been shown to be modular and can be swapped to change the effector recognition profile. The modularity of IDs has facilitated many attempts at engineering expanded effector recognition profiles to achieve novel disease resistance. However, this process is mostly limited to IDs where the effector-ID interface is already understood. Utilising de novo protein design tools, we have generated effector-binding proteins (binders) against three M. oryzae effectors: AVR-Pii, AVR-PikF, and APikL2. Here, we show that some of these binders interact with their cognate effector in vitro and trigger immune responses in planta when integrated as an ID within an NLR. Our work advances previous efforts to broaden NLR recognition by allowing the design of novel effector-binding IDs based on protein structure alone.

The poster QR code links to: https://github.com/ahbucknell/mpmi-poster