CDPK (calciumdependent protein kinase) gene was proved to play an important role in the biological and abiotic stress of plants. In this study, Dendrobium ‘Rainbowcompactum’ was used as the experimental material, calciumdependent protein kinase genes were cloned by RTPCR and their bioinformatics analysis were carried out. The expression of CDPKs gene in different tissues and under different low temperature stress was analyzed by qRTPCR, which laid the foundation for the functional verification of the CDPK gene and the research on the molecular mechanism of cold resistance of Dendrobium spp.. The results show that: (1) DenCDPK1 (GenBankaccession number: MZ322902), DenCDPK2 (GenBank accession number: MZ322903), DenCDPK3(GenBank accession numbe: rMZ322904) were cloned, with 1 934, 1 971 and 2 302 bp in length, encoding 534, 541 and 536 amino acids, respectively. (2) Sequence identity analyses showed that the identity of DenCDPKs protein and corresponding CDPK protein of Dendrobium officinale were both higher than 97%, and DenCDPKs contained a STKc_CAMK domain and 4 EFhand domains. (3) qRTPCR analysis showed that DenCDPK1, DenCDPK2 and DenCDPK3 were all highly expressed in leaves, and lowly expressed in stems, flowers and stems, respectively. The expression levels of DenCDPK1 and DenCDPK2 in leaves of Den. ‘Rainbowcompactum’ were higher than those of other three tested varieties. After being stressed at 8 ℃ for 12 h, the expression of DenCDPKs in leaves was significantly higher than that in the control group, and the expression of DenCDPK3 reached the highest value at 0 ℃ for 12 h. (4) The relative electrical conductivity in the leaves of Den. ‘Rainbowcompactum’ was lower than those of other three tested varieties under normal temperature (25 ℃). After low temperature (8 ℃, 0℃) stress for 12 h, the relative electrical conductivity in the leaves was significantly higher than that of the control group, and showed an upward trend with the decrease of temperature. It is speculated that DenCDPKs genes may be involved in the response to low temperature stress of Dendrobium spp., especially DenCDPK2 may play an important role in the cold resistance process of Dendrobium spp..