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MADS-box genes form a large gene family in plants and are involved in multiple biological processes, such as flowering. However, the regulation mechanism of MADS-box genes in flowering remains unresolved, especially under short-term cold conditions. In the present study, we isolated  BcMAF1 , a Pak-choi ( Brassica rapa  ssp.  Chinensis )  MADS AFFECTING FLOWERING  ( MAF ), as a floral repressor and functionally characterized  BcMAF1  in  Arabidopsis  and Pak-choi. Subcellular localization and sequence analysis indicated that BcMAF1 was a nuclear protein and contained a conserved MADS-box domain. Expression analysis revealed that  BcMAF1  had higher expression levels in leaves, stems, and petals, and could be induced by short-term cold conditions in Pak-choi. Overexpressing  BcMAF1  in  Arabidopsis  showed that  BcMAF1  had a negative function in regulating flowering, which was further confirmed by silencing endogenous  BcMAF1  in Pak-choi. In addition, qPCR results showed that  AtAP3  expression was reduced and  AtMAF2  expression was induced in  BcMAF1 -overexpressing  Arabidopsis . Meanwhile,  BcAP3  transcript was up-regulated and  BcMAF2  transcript was down-regulated in  BcMAF1 -silencing Pak-choi. Yeast one-hybrid and dual luciferase transient assays showed that BcMAF1 could bind to the promoters of  BcAP3  and  BcMAF2 . These results indicated that  BcAP3  and  BcMAF2  might be the targets of BcMAF1. Taken together, our results suggested that  BcMAF1  could negatively regulate flowering by directly activating  BcMAF2  and repressing  BcAP3 .

Isolation and Functional Characterization of a Floral Repressor, BcMAF1, From Pak-choi (Brassica rapa ssp. Chinensis)