BcFLC2 functioned as a repressor of flowering by directly regulating BcTEM1, BcMAF2, BcSOC1 and BcSPL15 in Pak-choi. FLOWERING LOCUS C (FLC) plays an important role in regulating flowering time. Here, we functionally described an FLC homologous gene, BcFLC2, that negatively regulated flowering in Pak-choi (Brassica rapa ssp. chinensis). The sequence comparison to Arabidopsis FLC showed that BcFLC2 also had a MADS-box domain at the N terminus. BcFLC2 was highly expressed in the leaves, roots, stems and stamens, and its expression was repressed by vernalization in Pak-choi. Interestingly, BcFLC2 expression exhibited a small peak at 2 weeks of vernalization treatment, suggesting that BcFLC2 may be involved in preventing premature flowering under short-term cold exposure in Pak-choi, which is different from the AtFLC expression pattern. Overexpression of BcFLC2 in Arabidopsis caused late flowering, while silencing of BcFLC2 in Pak-choi caused early flowering. BcFLC2 localized to the cell nucleus and functioned as a transcription factor. Yeast one-hybrid analysis revealed that BcFLC2 could bind to the promoters of Pak-choi Tempranillo 1 (BcTEM1), SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (BcSOC1), SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 15 (BcSPL15) and MADS AFFECTING FLOWERING 2 (BcMAF2). Taken together, the present results suggested that BcFLC2 played a key role in flowering regulation as a negative regulator by controlling BcTEM1, BcMAF2, BcSOC1 and BcSPL15 expression.
Keywords: FLOWERING LOCUS C 2; Flowering time; Late flowering; MADS AFFECTING FLOWERING 2; Short-term cold exposure; TEMPRANILLO 1; Vernalization.