The heterologous expression in Arabidopsis thaliana of a chrysanthemum gene encoding the BBX family transcription factor CmBBX13 delays flowering

Members of the B Box (BBX) family of proteins are known to be important for directing the growth and development of the Arabidopsis thaliana plant. Here, an analysis of a newly isolated chrysanthemum gene encoding a BBX family member implied that it was a likely ortholog of AtBBX13. The gene (designated CmBBX13) was most actively transcribed in the leaves and stem apex. CmBBX13 transcription was arrhythmic under either continuous darkness or continuous light, so the observed diurnal variation in its transcription appeared not to respond to the circadian clock. The outcome of transiently expressing CmBBX13 in onion epidermal cells suggested that the CmBBX13 protein localized to the nucleus. Both a yeast- and a protoplast-based assay showed that the protein has transactivational activity. When CmBBX13 was constitutively expressed in A. thaliana, flowering was delayed under both short and long day conditions. The presence of the transgene also downregulated a number of genes known to promote flowering, including APETALAI (API), SUPPRESSOR OF OVE-REXPRESSION OF CO 1 (SOC1), FLOWERING LOCUS T (PT) and FD, while simultaneously up-regulating the floral inhibitor-encoding genes FLOWERING LOCUS C (FLC) and TARGET OF EAT 2 (TOE2). The data suggested that CmBBX13 regulates flowering time independently of the photoperiod pathway.