Figure 4. BIK1 level accounts for G protein-mediated regulation of FLS2 immunity.

(A) Transient expression of BIK1 in agb1 and xlg2/3 mutant protoplasts restores RbohD phosphorylation. FLAG-RbohD and BIK1-HA constructs are transiently expressed in protoplasts from WT (Col-0), agb1 and xlg2/3. FLAG-RbohD protein was affinity purified and detected by anti-FLAG and anti-pSer39 immuoblots. (B) BIK1 transgene restores flg22-induced ROS burst in agb1. (C) BIK1 transgene partially restores flg22-induced resistance to Pst in agb1. NP::BIK1-HA was introduced into agb1 by crossing, transgenic lines of agb1 or Col-0 background in the F3 generation were used for the assays. (D) BIK1 transgene partially restores flg22-induced ROS burst in xlg2 xlg3 mutant. (E) BIK1 transgene partially restores flg22-induced resistance to Pst. The NP::BIK1-HA transgene was introduced into WT (Col-0-L32) and xlg2 xlg3 (xlg2/3-L64 and xlg2/3-L51) plants by Agrobacterium-mediated transformation. Independent T2 transgenic lines were used for the assays. Peak relative luminescence unit (RLU) values were shown for ROS assays (B and D) and leaf bacterial populations 2 days after bacterial inoculation were shown for flg22-protection assays (C and E). Bars in B-E represent mean ± SD (n ≥ 6; p<0.05, Student’s t-test; different letters indicate significant difference). Each experiment was repeated two (A) or three (B–E) times, and data of one representative experiment are shown.

DOI: http://dx.doi.org/10.7554/eLife.13568.016