Tetranychus cinnabarinus (Boisduval), i.e., carmine spider mite, is a worldwide pest that can cause serious damage to plants. Problems of resistance have arisen since abamectin usage in the control of T. cinnabarinus. Unfortunately, there are only limited data on the extent of this problem. To understand the development of abamectin resistance in the carmine spider mite, we prokaryotically expressed an intradiol ring-cleavage dioxygenase (ID-RCD) gene sequence, TcID-RCD1, which had a significant upregulated expression of over 7.7 times in an abamectin-resistant strain (AbR) when compared with that of a susceptible strain (SS). The crude enzyme activity also indicated that the AbR had a higher activity than that exhibited in SS. When susceptible individuals were treated with abamectin, TcID-RCD1 was also overexpressed. Furthermore, using the RNA interference (RNAi) technique, TcID-RCD1 was successfully knocked down, with the expression level decreasing significantly to approximately 39% in the SS strain compared with the control. And the mortality of mites feeding on dsTcID-RCD1 increased significantly when treated with abamectin. These results strongly suggest that TcID-RCD1 is involved in abamectin resistance in T. cinnabarinus.