Two B-box domain proteins, BBX28 and BBX29, regulate flowering time at low ambient temperature in Arabidopsis.

KEY MESSAGE: This paper demonstrates that BBX28 and BBX29 proteins in Arabidopsis promote flowering in association with the CO-FT regulatory module at low ambient temperature under LD conditions. Flowering plants integrate internal developmental signals with external environmental stimuli for precise flowering time control. The expression of BBX29 is up-regulated by low temperature treatment, but the biological function of BBX29 in low temperature response is unknown. In the current study, we examined the biological role of BBX29 and its close-related protein BBX28 in flowering time control under long-day conditions. Although neither BBX28 single mutant nor BBX29 single mutant has a flowering-associated phenotype, the bbx28 bbx29 double mutant plants have an obvious delayed flowering phenotype grown at low ambient temperature (16°C) compared to the wild-type (WT) plants. The expression of FT and TSF was lower in bbx28 bbx29 double mutant plants than in wild-type plants at 16°C. Both BBX28 and BBX29 interact with CONSTANS (CO), an important flowering integrator that directly binds to the FLOWERING LOCUS T (FT) promoter. In the effector-reporter assays, transcriptional activation activity of CO on the FT promoter was reduced in bbx28 bbx29 double mutant plants compared to that in WT plants. Taken together, our results reveal that BBX28 and BBX29 are promoters of flowering in Arabidopsis, especially at low ambient temperature.