Our study showed visible spatial and temporal patterns of amphibian and reptile diversity in Hupingshan National Nature Reserve. The patrolling and monitoring data from 2013 to 2016 was used to assess amphibian and reptile diversity of Hunan Hupingshan National Nature Reserve. In order to carry out routine monitoring, Hupingshan Reserve set 25 amphibian and reptile investigating transects at 8 monitoring stations, including 14 daytime transects with an average length of 4 km and 11 nighttime transects with an average length of 0.75 km (Fig. 1). Each transect was investigated 3 times per month on average. During these 4 years, 62 amphibian and reptile species were recorded in total, accounting for 38.04% of the amphibian and reptilian species in Hunan province. We calculated the alpha diversity index of each monitoring site and the beta diversity index of the whole reserve monthly and compared the Hellinger distance of species composition in different stations among seasons. And we respectively applied ANOVA to analyze significances. The results showed that there was no significant difference among amphibian and reptile diversity in different locations, monitoring stations and in different years, which means the ecological environment of reserve is relatively stable. However, species composition fluctuated drastically in several stations located in or near experimental zone of reserve, which means human activities are influential to the distribution of amphibian and reptilian species. Besides, there were significant seasonal differences in species composition and diversity of amphibians and reptiles in the reserve, which is related to the biological characteristics of amphibians and reptiles. The results above indicate that the diversity structure of amphibians and reptiles in Hupingshan Reserve is stable yet sensitive, which can be considered as a monitoring index for the biodiversity level in the reserve.