DREB gene is called the main regulator of abiotic stress response. In order to explore the function of DREB in the stress resistance process of Dryopteris fragrans (L.) Schott, a perennial medicinal herb, this study cloned and analyzed the DfDREB gene with bioinformatics. The expression patterns of DfDREB gene under different hormones and stress treatments of drought, NaCl, high temperature and low temperature were studied by realtime quantitative polymerase chain reaction. The results showed that： (1) the DfDREB gene, with a total length of 1 203 bp, encodes 401 amino acids. The relative molecular mass is 43.66 kD and the isoelectric point is 6.13, which is a hydrophilic nonsecreted protein. This protein has an AP2 conserved domain and belongs to the AP2 family. (2) Realtime PCR analysis showed that DfDREB gene was expressed in the roots, petioles and leaves of D. fragrans, among which the highest expression was in the leaves and the lowest expression was in the roots. When SA, MeJA and ETH were treated, the expression of DfDREB was upregulated, and the expression reached the peak at 1 h. During ABA treatment, the relative expression of DfDREB was upregulated only at 12 and 24 h, and downregulated during the rest of the time. Under drought, NaCl and high temperature treatments, DfDREB expression was upregulated. The expression of DfDREB was significantly upregulated at 0.5 and 12 h under low temperature treatment, and there was no significant change at 1 to 6 h and 24 h. It shows that DfDREB can respond to hormone and abiotic stress treatments, and gene expression is induced by stress. The results provide a scientific basis for further study on the stress resistance mechanism of D. fragrans.