Abstract: CBFs (CRT/DRE-binding factor) are key transcription factors regulating the chilling response in plants. A 457 bp CBF-like fragment was cloned from Jacaranda acutifolia. Sequence analysis showed that the fragment (named as JaCBF1) encoded a protein of 142 amino acids, and was highly homologous to many known CBF1-like transcription factors with the AP2 core domain that belongs to CBF gene family. The recombinant plasmid containing JaCBF1 sequence was introduced into Escherichia coli. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that JaCBF1 was translated into a 16 KDa protein. Expression pattern analysis indicated that JaCBF1 could be induced with different patterns and degrees by low temperature in roots, stems, and leaves of J. acutifolia. Southern blot revealed that the J. acutifolia genome contained at least two copies of JaCBF1. Ectopic transcription in Arabidopsis thaliana revealed that JaCBF1 functioned in the nuclei, and its expression could significantly improve the freezing resistance of transgenic A. thaliana. The results of study suggest that the novel JaCBF1 is a typical CBF-like transcription factor, and plays an important role responding to cold stress in J.acutifolia.