Fig. 5
The SOS domain is essential for EX1 degradation and 1O2 signaling. a EX1ΔSOS-GFP, EX1F528C-GFP, and EX1G646D-GFP are unable to complement ex1 flu. Seedlings of WT, flu, ex1 flu, EX1-GFP ex1 flu (EX1), EX1ΔSOS-GFP ex1 flu (ΔSOS), EX1F528C-GFP ex1 flu (F528C), and EX1G646D-GFP ex1 flu (G646D) grown for 5 days under CL were shifted to LD conditions for 3 days. Seedlings grown under CL for 8 days serve as controls. b Pchlide accumulation in etiolated seedlings. c Singlet oxygen sensor green (SOSG) fluorescence and d expression of SORGs were determined as shown in Fig. 3c, d. ACT2 was used as an internal control. Data represents the mean of three independent biological replicates. Error bars indicate standard deviation. Lower case letters indicate statistically significant differences between the mean values (P < 0.05, one-way analysis of variance with post hoc Tukey’s Honest Significant Difference test). e Deletion or modification of the SOS domain prevents the EX1 degradation. Total protein isolated from 5-day-old seedlings initially grown under CL and then subjected to 8 h dark followed by 30-, 60-, and 120-min re-illumination, respectively, were analyzed by western blot using GFP antibody. UGP was used as a loading control
