Fig. 3. Engineering Agrobacterium flg22 perception by residue transfer from FLS2^XL.

a, AlphaFold3 model of the LRR extracellular domains of FLS2^XL and co-receptor NbSERK3A with Atu flg22 peptide. Colour represents pLDDT. b, RCM of 26 FLS2 homologues in the Vitales order, showing conservation levels from high (yellow) to low (dark blue). White asterisks and bars mark the residues transferred from FLS2^XL to VrFLS2 in synthetic VrFLS2 (SynFcFLS2^22XL), which are associated with the altered ligand detection capability. Colour bars indicate the interacting regions of flg22 ligand or co-receptor SERK3. c, Representations of FLS2 variants. Colour bars in the LRRs indicate the occurrence of residue transfers. The numbers of residues transferred are superscripted. d, Normalized ROS production after treatment with flg22 variants (100 nM). FLS2s were transiently expressed in the N. benthamiana fls2-1/2 CRISPR–Cas9 mutant. Normalized ROS levels were calculated by adjusting maximum RLU averages to a 0 (water) to 100 (Pae flg22) scale. Data points below a value of 10 are depicted as white bubbles. e,f, Top view (e) and binding interface (f) of AlphaFold3 model showing the LRR extracellular domain of SynFcFLS2^22XL and co-receptor NbSERK3A with Atu flg22 peptide. The N and C termini of flg22 are labelled. g, Phosphorylation of MAPK induced by flg22 variants (100 nM) for different receptors. CBB staining indicates protein loading. All experiments were repeated independently at least three times with similar results. Vr, Vitis riparia.

Source data