Arabidopsis ELF4-like proteins EFL1 and EFL3 influence flowering time

The circadian clock synchronizes internal and external stimuli to ensure numerous biological processes occur at the optimal time. EARLY FLOWERING 4 (ELF4) is a key evening-phased component of the circadian clock and essential for photoperiod-dependent flowering time regulation in Arabidopsis thaliana. There are four homologous ELF4-like (EFL1-EFL4) genes in the Arabidopsis genome but their functions are unknown. Protein sequence alignment and phylogenetic analysis showed that these four EFL proteins contained an evolutionarily conserved domain, DUF1313, of unknown function. To investigate the physical roles of these genes in Arabidopsis, we overexpressed the four homologous EFL genes in the elf4 mutant background. Under both long-day (LD) and short-day (SD) conditions, overexpression of EFL1 not only completely rescued the early flowering phenotype of the elf4 mutant, but also delayed flowering. Overexpression of EFL2, however, failed to rescue this phenotype and overexpression of EFL3 partially rescued the early flowering phenotype. The transcription levels of the key flowering time regulation genes CONSTANS (CO) and FLOWERING LOCUS T (FT) were significantly decreased in the EFL1- and EFL3-overexpressing transgenic lines in a dose-dependent manner, compared with the elf4 mutant. These results suggest that EFL1 and EFL3 are involved in flowering time regulation in Arabidopsis.