Sophisticated social behaviors in termite colonies are mainly regulated via chemical communication of a wide range of pheromones. Trail pheromones play important roles in foraging behavior and building tunnels and nests in termites. However, it is almost unclear how termites perceive trail pheromones. Here, we cloned and sequenced of olfactory co-receptor (Orco) genes from the two termites Reticulitermes chinensis Snyder (Isoptera: Rhinotermitidae) and Odontotermes formosanus (Shiraki) (Isoptera:Termitidae), and then examined their responses to trail pheromones after silencing Orco through RNA interference (RNAi). We found that Orco knockdown impaired their ability to perceive trail pheromones and resulted in the disability of following pheromone trails in the two termite species. Our locomotion behavior assays further showed that Orco knockdown significantly decreased the distance and velocity in the two termite species, but significantly increased the angular velocity and turn angle in the termite R. chinensis. These findings strongly demonstrated that Orco is essential for termites to perceive their trail pheromones, which provides a potential way to control termite pests by damaging olfactory system.