The two spotted mite, Tetranychus urticae Koch, is a global pest, and has developed severe resistance to several types of acaricides. Acequinocyl-resistant strain and bifenazate-resistant strain of T. urticae showed higher relative activity level of glutathion-S-transferases (GSTs) than susceptible strain. Studies of resistance in T. urticae have focused on classical detoxifying enzyme system, such as GSTs. We examined the expression of six GSTs genes in T. urticae belong to five different cytosolic classes, including two in delta, one in omega, one in mu, one in zeta, and one mitochondrial GSTs class, kappa. RT-qPCR results showed that, among these six GSTs genes, the relative expression level of delta 1 was significantly higher in bifenazate-resistant strain than in other strains (susceptible, abamectin, acequinocyl, etoxazole and pyridaben strain). Expression level of the delta 2 was also higher in acequinocyl-resistant strain than in other strains. The elevated GSTs gene transcripts following exposure to acequinocyl, bifenazate might be one of the mechanisms involved in detoxification of these acaricide. These results may be caused by different mechanisms in the resistance development of T. urticae to these two different types of acaricides.