Among the community of vertebrates living in the natural environment, reptiles are more prone to be affected by various road conditions like vehicle collisions which severely threaten their life. From 2019 to 2020, road driving survey was conducted on the roadkill of reptiles to describe road-kill composition and to estimate the main reasons of road-kills in Simao District, Pu'er City, Yunnan Province. The results showed that there were recorded 100 carcasses of reptiles belonging to 22 species within 7 families, among them Pareas margaritophorus of Pareatidae and Bungarus muliticinctus of Elapidae were more vulnerable to road mortality. It was also notable that two species of State Key Protected Wild Animal Species, Xenopeltis unicolor and Elaphe radiatus, were also been found in roadkill in this case. There were three peaks of reptile roadkill, 76 carcasses of reptiles recorded at 7:00-10:00, probably due to their higher activity, which was the hot moment of roadkill. The temperature ranges of reptiles roadkill were concentrated between 20-25℃. Furthermore, considering the physiological needs of reptile, such as thermoregulation, reptiles roadkill were coincident with their activity rhythm, especially during the peak period of basking between 7:00-10:00, which made them more prone to collisions.