Extended Data Fig. 5. Engineering FLS2^XL specificity.

a, Normalized ROS induction of flg22 variants (100 nM) in N. benthamiana fls2-1/2 expressing FLS2^XL, VrFLS2, and synthetic VrFLS2s. Each data point represents an average max relative light unit (RLU) from 4 plants with 4 leaf disks per plant. The values of each plate were normalized by adjusting maximum relative light unit (RLU) averages to a 0 (water) to 100000 (Pae) scale. One-way ANOVA and Dunnett’s multiple comparison were used to determine significance using the non-scaled data, p values are marked next to each bar. Error bars = SEM. b, Protein sequence alignment for LRR domains of five FLS2 variants. Concave surface regions of each LRR are highlighted with dark bars. Residues on the interacting surface are determined using the AlphaFold3 model of FLS2^XL/NbSERK3A/Atuflg22 (< 5 Å). Residues in contact with flg22 are highlighted in cyan above the alignment, while residues in contact with the co-receptor SERK3 are highlighted in purple. Residues swapped for SynVrFLS2^22XL are highlighted with red blocks.