The caleosins are encoded by multi-gene families in Arabidopsis thaliana and other plant species. This work investigates the role of two family members, RD20/CLO3 and CLO7, in flowering transition and in root development in response to ABA treatment. Gene expression of the caleosin RD20/CLO3 is induced by ABA in the root tissues and RD20/CLO3 has a negative affect on the total number of lateral roots as well as the length of the lateral roots in response to ABA treatment. The rd20/clo3 mutant has more and longer lateral roots in response to ABA treatment compared to the wild-type, showing that RD20/CLO3 plays a role in the ABA signaling pathway affecting this trait. In contrast, the caleosin CLO7 is not expressed in the roots and does not affect root architecture in response to ABA treatment. The disruption of both RD20/CLO3 and CLO7 together causes a dramatic early-flowering phenotype under long-day conditions, whereas single mutations in these genes do not affect flowering time under these conditions. Both yeast two-hybrid and bimolecular fluorescence complementation showed that both RD20/CLO3 and CLO7 interact with each other and can form homodimers and heterodimers. Taken together, these findings suggest that members of the caleosin gene family play both different and redundant roles in plant development.