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Concurrent Session

Session: Concurrent Session: Structure Informed Synthetic Biology: From Design to Function

Pikobodies: NLR immune receptor-nanobody fusions can confer resistance to the potato blight pathogen Phytophthora infestans

Thursday, July 17, 2025
16:35 - 16:45  
Location: Confex Hall 1+2

Plant pathogens continuously evolve to evade immunity, driving the need for innovative resistance strategies to safeguard global food security. In proof-of-concept work, we engineered the rice nucleotide-binding domain leucine-rich repeat receptor (NLR) pair Pik-1/Pik-2 to detect fluorescent proteins (FPs) (Kourelis, Marchal et al. PMID 36862785). We achieved this replacing the Pik-1 integrated HMA domain with nanobodies binding the fluorescent proteins GFP or mCherry, naming these bioengineered receptor-nanobody fusions Pikobodies. We previously showed that Pikobodies confer resistance to potato virus X variants expressing FPs. Given nanobody versatility to bind virtually any antigen, we hypothesized that Pikobodies can be engineered to recognize intracellular pathogen effectors, enabling made-to-order disease resistance genes. Here we generated Pikobodies carrying nanobodies binding the RXLR-LWY effector AVRcap1b, secreted by the potato blight pathogen Phytophthora infestans. We tested nanobody fusions in the Pik-1 scaffold in Nicotiana benthamiana for compatibility with Pik-1 and specificity to AVRcap1b (i.e. cell-death induction exclusively upon effector presence). Stable transgenic N. benthamiana lines expressing anti-AVRcap1b Pikobodies exhibited resistance to P. infestans comparable to potato blight resistance gene Rpi-blb2. In light of this, we discuss challenges and opportunities of the Pikobodies technology for advancing bespoke plant disease resistance engineering.