Increased freezing tolerance through up-regulation of downstream genes via the wheat CBF gene in transgenic tobacco

The wheat (Triticum aestivum L.) CBF gene family is assumed to play important roles in development of low-temperature and freezing tolerance through activation of the downstream Cor/Lea genes. However, no direct evidence shows association of the wheat CBF genes with stress tolerance or any interaction between wheat CBF transcription factors and Cor/Lea gene activation. Here, we introduced Wcbf2, one of the wheat CBF genes, into the tobacco (Nicotiana tabacum L.) genome. Expression of Wcbf2 significantly increased the level of freezing tolerance in the transgenic tobacco plants without phenotypic retardation, and altered the expression patterns of tobacco genes, including cold-responsive genes. A transgenic tobacco plant expressing Wcbf2 was crossed to other transgenic plants expressing a GUS reporter gene under control of the wheat Cor/Lea gene promoter. Analysis of the F1 plants showed that the WCBF2 protein positively regulated at least the expression of Wdhn13 and Wrab17. These results strongly indicate that WCBF2 functions as a transcription factor in the development of freezing tolerance in common wheat.