With the recent increase in planting of fruit trees in southern Xinjiang, the intercropping of fruit trees and cotton has been widely adopted. From 2014 to 2016, a large-scale study was conducted in Aksu, an important agricultural area in southern Xinjiang, to compare the abundance and species composition of spider mites in cotton fields under jujube-cotton, apple-cotton, and cotton monocrop systems. The abundance of spider mites in cotton fields under both intercropping systems was generally higher than in the cotton monocrop. The species composition of spider mites also differed greatly between cotton intercropped with apple or jujube compared to the cotton monocrop. The relative proportion of Tetranychus truncates Ehara (Acari: Tetranychidae) in the species complex generally increased while that of another spider mite, Tetranychus dunhuangensis Wang (Acari: Tetranychidae), decreased under fruit tree-cotton systems. More attention should be paid to the monitoring and management of spider mites, especially T. truncates in this important region of China.