It has been proved that agroinfiltration-based temporary expression of coatomer subunit alpha (COPA) gene from Tetranychus urticae hairpin RNA induces RNA interference (RNAi) and lethality to T. urticae. To establish detailed protocols for agroinfiltration, the efficiency of agroinfiltration to the soybean and kidney bean was determined with respect to different Agrobacterium delivery methods (sea sand, carborundum and syringe) and the spacial expression patterns of hairpin RNA was investigated following Agrobacterium delivery. Sea sand and syringe showed the highest expression level in soybean and kidney bean, respectively. Considering the resulting tissue damage, syringe appeared the best choice for agroinfiltration in both soybean and kidney bean. The apical region of a leaf showed more relative expression levels in both soybean and kidney bean compared to the basal region. Following agroinfiltration, adjacent untreated leaves were determined to express hairpin RNA though the expression level was low, suggesting that hairpin RNA can be translocated to other leaves. In conclusion, Agrobacterium delivery by syringe and use of whole leaf were recommended for T. urticae bioassay following agroinfiltration.