Knocking out SOBIR1 in Nicotiana benthamiana abolishes functionality of transgenic receptor-like protein Cf-4

The first layer of plant immunity is formed by pattern recognition receptors (PRRs) that are present at the cell surface and perceive extracellular immunogenic patterns. Receptor-like proteins (RLPs), such as the tomato ( Solanum lycopersicum ) PRR Cf-4 that provides resistance to the fungus Cladosporium fulvum secreting the matching avirulence factor Avr4, have an extracellular receptor domain consisting of leucine-rich repeats, but lack a cytoplasmic kinase domain for downstream signaling. RLPs constitutively interact with the receptor-like kinase SUPPRESSOR OF BIR1-1 (SOBIR1), thereby providing the receptor with a kinase domain, and recruit the co-receptor BRI-ASSOCIATED KINASE 1 (BAK1) upon their activation by a matching ligand. Trans-phosphorylation events, which can take place between the kinase domains of SOBIR1 and BAK1 after their association with the RLP, are thought to initiate downstream defense signaling. Currently, our knowledge on RLP/SOBIR1/BAK1-mediated defence initiation is limited and to understand the role of SOBIR1 in RLP function, we knocked out SOBIR1 and its close homolog SOBIR1-like in the model plant Nicotiana benthamiana , as well as in transgenic N. benthamiana stably expressing Cf-4 . We observed that Cf-4 function is completely abolished in the knock-out mutants, and we show that these plants can be used to perform transient complementation studies with SOBIR1 mutants. Thereby, these mutants are an important tool to study the fundamentals of plant immunity mediated by RLPs. ### Competing Interest Statement The authors have declared no competing interest.