Abstract: To safely and effectively prevent and control tobacco target spot disease, endo polygalacturonases (endoPGs) and MAP kinases of the tobacco target spot pathogen (Rhizoctonia solani YC-9)were used as the targets to design and synthesize dsRNA-endoPGs and dsRNA-RMK1 by RNA interference technology. The control effects of dsRNA-endoPGs and dsRNA-RMK1 on tobacco target spot were analyzed by spraying on tobacco leaves in vitro and tobacco plants in the field. The results showed that 7 d after spraying dsRNA-endoPGs and dsRNA-RMK1 on tobacco leaves in vitro, the inhibition rates of tobacco target spot lesions were 62.10% and 60.00%, respectively. After spraying dsRNA-endoPGs or dsRNA-RMK1 on the leaves of tobacco plants in field trials once every 7 d and 3 times in total, the relative control efficacies on tobacco target spot 7, 14, 21 d after the first spray were all above 50%.