The screening of effective lethal genes was conducted via the systemic delivery of dsRNA for the RNAi-based management of Tetranychus urticae. Six candidate genes (COPI coatmer, T_COPI; ESCRT III_Snf7, T_SNF7; Ribosomal protein S4, T_RPS4; v-ATPase A subunit 2, T_V-ATPase; Aminopeptidase N, T_APN3; Acetylcholinesterase, T_AChE) and two reference genes (EGFP and T_AChEintron) were tested for the experiment. The permeated dsRNA to the leaf disc (ca. 30 mm diameter) was detected at 6 h after treatment, indicating that dsRNA could move through veins on the leaf. In the reference gene selection, the T_AChEintron was chosen for its low mortality compared with EGFP gene. In the evaluation of mortality, the T_COPI, T_V-ATPase and T_RPS4 exerted higher toxicities at 24 hour after treatment among six genes tested. Interestingly, T_APN3 showed toxicity after 72 hour. In summary, the dsRNA delivery via leaf disc was effective in screening lethal genes and some genes, such as COPI, V-ATPase and RPS4, can be applicable for establishing a RNAi-based control system against T. urticae.