BLISTER-regulated vegetative growth is dependent on the protein kinase domain of ER stress modulator IRE1A in Arabidopsis thaliana
Fig 5
Residues in the protein kinase domain of IRE1A is required for the shoot growth phenotype in BLI mutant plant background.
A-B, Site-specific mutagenesis analysis of the IRE1A auto-phosphorylation. Domain structure of IRE1A is shown in (A) with the mutated amino acids highlighted. The wild-type form and various mutated forms of IRE1A were expressed in E. coli and purified proteins were treated with CIAP for de-phosphorylation assays. Western blotting analysis was done to detect the molecular weight shifts. C-D, Genetic complementation analysis. T-DNA mutant bli (bli-1) was crossed to either IRE1A mutant (ire1a) or IRE1B mutant (ire1b) to generate the respective double mutant plants. The wild-type form and various mutated forms of IRE1A were expressed the bli ire1a double mutant background and 2-weeks-old T3 transgenic plants were photographed (C) and fresh weight were measured (D). Error bars represent SE (n = 3). There were at least 10 plants for each of the three biological replicates. Letters above the bars indicate significant differences as determined by LSD test following ANOVA analysis (p<0.05). Bar = 5 mm.