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Background  ICE (Inducer of  CBF  Expression) family genes play an important role in the regulation of cold tolerance pathways. In an earlier study, we isolated the gene  CdICE1  from  Chrysanthemum dichrum  and demonstrated that freezing tolerance was enhanced by  CdICE1  overexpression. Therefore, we sought to determine the mechanism by which  ICE1  family genes participate in freezing tolerance.    Results  Using EMSA (Electrophoretic Mobility Shift Assay) and yeast one-hybrid assays, we confirmed that CdICE1 binds specifically to the MYC element in the  CdDREBa  promoter and activates transcription. In addition, overexpression of  CdICE1  enhanced  Arabidopsis  freezing tolerance after transition from 23°C to 4°C or 16°C. We found that after acclimation to 4°C, CdICE1, like  Arabidopsis  AtICE1, promoted expression of  CBF s ( CRT/DRE Binding Factor ) and their genes downstream involved in freezing tolerance, including  COR15a  ( Cold-Regulated 15a ),  COR6.6 , and  RD29a  ( Responsive to Dessication 29a ). Interestingly, we observed that  CdICE1 -overexpressing plants experienced significant reduction in miR398. In addition, its target genes  CSD1  ( Copper/zinc Superoxide Dismutase 1 ) and  CSD2  showed inducible expression under acclimation at 16°C, indicating that the miR398-CSD pathway was involved in the induction of freezing tolerance.    Conclusions  Our data indicate that  CdICE1 -mediated freezing tolerance occurs via different pathways, involving either CBF or miR398, under acclimation at two different temperatures.

Ambient temperature enhanced freezing tolerance of Chrysanthemum dichrum CdICE1Arabidopsis via miR398