Osmanthus fragransis a popular ornamental and odorant plant with high commercial value, but its cultivation and exploitation are limited by low temperature. TheZAT(zinc finger ofArabidopsis thaliana) genes as a subclass of the C2H2-type zinc finger proteins (C2H2-ZFP) family play essential roles in various abiotic stresses. However, their roles in cold stress response inO. fragransremain unclear. This study identified 38OfZATs, which could be divided into 5 subgroups based on the phylogenetic tree, withOfZATsin the same subgroup harboring similar gene structures and motif patterns. In addition, 49 segmental and 5 tandem duplication events were detected amongOfZATgenes, while someOfZATgenes exhibited specific expression patterns in different tissues. Furthermore, twoOfZATs were induced in salt stress and eightOfZATsresponded to cold stress. Interestingly,OfZAT35showed a continuously increasing expression trend under cold stress, while its protein showed nucleus localization with no transcriptional activation activity. Transiently transformed tobacco overexpressingOfZAT35exhibited a significantly higher relative electrolyte leakage (REL) level and increased activities of superoxide dismutase (SOD), peroxidase (POD), and Ascorbate peroxidase (APX), while there was significantly decreased activity of catalase (CAT). Moreover,CAT,DREB3, andLEA5, which are associated with cold stress, were dramatically decreased after cold treatment in transiently transformed tobacco, suggesting that overexpression ofOfZAT35negatively regulated cold stress. This study provides a basis for exploring the roles ofZATgenes and contributes to uncovering the mechanism ofZAT-mediated cold stress response inO. fragrans.