Extended Data Fig. 3: Characterization of polymorphic residues in the engineered variant ‘V2’.
Flg22Pa- and flg22Atum-induced ROS burst kinetics (a) and MAPK phosphorylation (b), inhibition of A. tumefaciens-mediated T-DNA transfer (c), and suppression of A. tumefaciens growth (d) by engineered variant ‘V2’ carrying single or multiple mutations at polymorphic sites (related to Fig. 2h). Number of leaf discs measured in a are indicated in the plots (nV2 and nmutant). For each FLS2 variant, flg22Pa and flg22Atum treatment groups have the same sample sizes. Samples sizes of pairs in c: n = 7, 15, 11, 12, 16, 6, 15, 15, 12, 9, 12, 9, 15, 12, 15 biological replicates, from left to right. Samples sizes of pairs in d: n = 32 leaf discs for ‘391,413,419’; n = 28 leaf discs for ‘S438G’, ‘F439L’, ‘K441V’; n = 24 leaf discs for the rest. For line charts and bar plots, data are shown as mean ± SEM. For box plots: center line, median; box limits, upper and lower quartiles; whiskers, 1.5 times interquartile range; points, outliers; ‘+’, mean. The dotted line indicates Y = 1. Two-tailed unpaired t test was used for mutants ‘A340V’, ‘D368S’, ‘L391V’, ‘V412N’, ‘L413I’, ‘I435T’, ‘F439L’, and ‘R465K’ in d. Two-tailed Mann–Whitney test was used for statistical analysis in c, and for mutants ‘Q344H’, ‘L345S’, ‘Y364I’, ‘N392H’, ‘S438G’, ‘K441V’, and ‘391,413,419’ in d. Similar results were obtained from two (b, c, d) or three (a) independent experiments.
