Fig. 2: OsLIKE1 phosphorylates LHCB5 enhancing rice resistance against M. oryzae.

a OsLIKE1-KD phosphorylates LHCB5. A kinase assay was performed using OsLIKE1-KD-His as the kinase and GST-LHCB5 as the substrate, simultaneously detecting the autophosphorylation of OsLIKE1-KD and phosphorylation of LHCB5. Protein OsLIKE1-KD and LHCB5 were subjected to Mn²⁺-Phos-tag SDS-PAGE and normal SDS-PAGE followed by immunoblotting with anti-His and anti-GST antibodies. b Blast resistance of OsLIKE1-OE and OsLIKE1-KO plants following Guy11 spore spraying. Diseased leaves were photographed after 7 days. c The leaf lesion area of the leaves from (b) was assessed by Image J. The experiments were repeated twice with similar results. Values are the means of 3 replications, and error bars rep resent the SD (n = 3). Data are analyzed by two-sided ANOVA followed by Tukey’s test. Adjusted p values are shown in figures. d The relative fungal biomass of the leaves from (b) was detected by qRT-PCR. Values are the means of 3 replications, and error bars rep resent the SD (n = 3). Data are analyzed by two-sided ANOVA followed by Tukey’s test. Adjusted p values are shown in figures. e Guy11 infection is required for OsLIKE1 to phosphorylate LHCB5. Total proteins from OsLIKE1-OE and OsLIKE1-KO plants, sprayed with Guy11 spores and incubated for 2 days, were subjected to Mn²⁺-Phos-tag SDS-PAGE and regular SDS-PAGE, followed by immunoblotting with anti-GST and anti-LHCB5 antibodies. f OsLIKE1 is necessary for the phosphorylation of LHCB5. The LHCB5-HA fusion protein was expressed in rice protoplasts of ZH11, OsLIKE1-OE and OsLIKE1-KO, respectively. The LHCB5-HA fusion protein was subjected to Mn²⁺-Phos-tag SDS-PAGE and normal SDS-PAGE followed by immunoblotting with an anti-HA antibody. All experiments were repeated independently with similar results at least three times.