Figure 1.

CML41 is induced by flg22 in leaves and binds Ca²⁺. (a) GUS histochemical staining of 24‐d‐old proCML41::GUS plants treated with either H₂O or flg22 infiltration for 4 h, as well as nontreatment control as indicated. Both flg22 and H₂O injection at the wound/infiltration site is indicated by arrowheads, there was a localized increase in GUS activity induced by both flg22 and H₂O injection at the wound site. (b) Quantitative RT‐PCR analysis of CML41 in the leaves of 5–6‐wk‐old wildtype Arabidopsis Col‐0 plants grown in short‐day conditions (with 9 h : 15 h, light : dark) pre‐infiltrated with either H₂O (green) or 1 μM flg22 (magenta) for 12 h. Gene transcript level was relative to GAPDH‐A (At3g26650). Data represent the mean ± standard error of the mean (SEM), n = 3 biological replicates. Primer pairs used for (b) listed in Supporting Information Table S1. Statistical difference was determined by Student's t‐test, asterisks indicate statistical significance, P‐value as indicated. (c) Gel shift Ca²⁺ binding assay, purified recombinant MBP‐CML41 protein was separated on 8% SDS‐PAGE gel in the presence of 1 mM CaCl₂ or 10 mM EGTA, the mobility of proteins was determined by comparison with the Precision Plus Protein™ Standards as indicated.