The C-repeat binding factors (CBFs) gene is essential for plants’ cold response, which could not only be induced by the inducer of CBF expression (ICE) genes but also activated the expression of the cold-regulated (COR) gene, thereby participating in the ICE-CBF-COR cold response pathway. However, this gene family and its functions in Actinidia arguta remain unclear. In this study, whole-genome identification and functional analysis of CBF family members in A. arguta were performed. Eighteen CBF genes, which were located on four chromosomes and had five tandem repeats, were identified. The proteins encoded by the genes were predicted to be located in the nucleus and cytoplasm. The results of the promoter cis-acting element analysis revealed light response elements, low-temperature response elements, and hormone (methyl jasmonate, gibberellin, salicylic acid, etc.) response elements. We analyzed collinearity with other kiwifruit genomes, and, interestingly, the number of CBF family members differed across geographic locations of A. arguta. RT-qPCR revealed that the expression of the CBF gene family differed under low-temperature treatment; specifically, we observed differences in the expression of all the genes. Based on phylogenetic relationships and RT-qPCR analysis, the expression of AaCBF4.1 (AaCBF4) was found to be highly upregulated, and the function of this gene in cold resistance was further verified via overexpression in transgenic Arabidopsis. AaCBF4-overexpressing plants showed higher tolerance to cold stress, showing a higher germination rate, higher chlorophyll content and lower relative electrolyte leakage. In addition, compared with the wild-type Arabidopsis, the overexpressing plants exhibited significantly reduced oxidative damage due to the reduction in reactive oxygen species production under cold stress. Therefore, AaCBF4 plays an important role in improving the cold resistance of Actinidia arguta and can be further used to develop kiwifruit germplasm resources with strong cold resistance.

Cold stress is one of the main abiotic stresses that affect the development and growth of kiwifruit (Actinidia arguta). Herein, we analyzed the transcriptomic data of A. arguta dormant shoots in response to low-temperature treatment, identified 52 MYB genes, and constructed a phylogenetic tree based on the encoded protein sequences. Then, the effect of one MYB gene on cold tolerance was analyzed. This gene had an open reading frame of 837 bp long and encoded 279 amino acids. Sequence alignment and phylogenetic analysis revealed that this gene belongs to the R2R3-MYB family and was named AaMYB44 based on its homology to other MYB family members. Quantitative real-time PCR revealed that AaMYB44 expression was significantly induced by low temperatures but exhibited the opposite trend in cold-tolerant genotypes. Subcellular localization assays revealed the nuclear localization of the AaMYB44 protein. Furthermore, AaMYB44 was transformed into Arabidopsis thaliana (A. thaliana) via inflorescence infection, and physiological and biochemical tests revealed that the cold resistance and antioxidant capacity of the transgenic A. thaliana were lower than those of wild-type plants. Overall, AaMYB44 might play a negative regulatory role in response to cold stress, providing new insight into the mechanism of cold tolerance.