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The C-REPEAT BINDING FACTOR signaling pathway is strictly modulated by numerous factors and is essential in the cold response of plants. Here, we show that the DUF793 family gene  BYPASS1-LIKE  modulates freezing tolerance through the CBFs in  Arabidopsis . The expression of  B1L  was rapidly induced under cold treatment. Comparing to wild type,  B1L  knockout mutants were more sensitive to freezing treatment, whereas B1L-overexpressing lines were more tolerant. The expression of  CBF s and CBF target genes was significantly decreased in  b1l  mutant. Using yeast two-hybrid screening system, 14-3-3λ was identified as one of proteins interacting with B1L. The interaction was confirmed with bimolecular fluorescence complementation assay and co-immunoprecipitation assay. Biochemical assays revealed that  b1l  mutation promoted the degradation of CBF3 compared to wild type, whereas  14-3-3κλ  mutant and  b1l 14-3-3κλ  mutant suppressed the degradation of CBF3. Consistently,  14-3-3κλ  and  b1l 14-3-3κλ  mutants showed enhanced freezing tolerance compared to wild type. These results indicate that B1L enhances the freezing tolerance of plants, at least partly through stabilizing CBF. Our findings improve our understanding of the regulation of CBF in response to cold stress.

BYPASS1-LIKE, A DUF793 Family Protein, Participates in Freezing Tolerance via the CBF Pathway in Arabidopsis