Extended Data Fig. 2: Characterization of engineered VvFLS2 variants.
a, Flg22Pa- and flg22Atum-induced ROS production of engineered FLS2 variants listed in Fig. 2a (corresponding to Fig. 2d, n = 64, 61, 56, 60, 84, 80, 40, 39 leaf discs, from top to bottom). b, Comparison of total flg22Pa-induced ROS production among engineered VvFLS2 variants (n = 64, 56, 84, 40 leaf discs). c, Restriction of A. tumefaciens growth by engineered VvFLS2 variants (n = 48 leaf discs), with bioluminescence of the ‘AgroLux’ strain used as a proxy for bacterial population size. d, Protein expression levels of GFP-tagged VvFLS2, VrFLS2XL, and ‘V2’. e, Effects of mutating clusters of polymorphic residues in each LRR from LRR12-17 of ‘V2’ (n = 24, 24, 20, 48, 12, 36 leaf discs from group ‘LRR12’ to group ‘LRR17’). f, Sequence alignment of the LRR12-19 region of VvFLS2, VrFLS2XL, and other previously reported flg22Atum-responsive FLS2 variants. Polymorphic sites between VvFLS2 and VrFLS2XL are marked with their position within VrFLS2XL. Polymorphic residue numbers in ‘V2’ variant are shown in bold. The legends are consistent with Fig. 2a. LRR annotations are based on ‘4MN8’ structure from the Protein Data Bank (PDB). (Qv, Quercus variabilis; Tj, Trachelospermum jasminoides; Sb, Salix babylonica; Pt, Populus trichocarpa). For line charts, data are presented as mean ± SEM. For box plots: center line, median; box limits, upper and lower quartiles; whiskers, 1.5 times interquartile range; points, outliers. Kruskal-Wallis test followed by Dunn’s multiple comparisons test was used for statistical analysis in b and c. Significant differences at P < 0.05 are indicated by letters. See Source data for exact P values. Sequence alignment and visualization were performed using Jalview69. All experiments were repeated three times with similar results, except for c, which was repeated twice.
