The Luoxiao Mountains play an important role in maintaining and supplementing the fish diversity of the Yangtze River Basin, which is also a biodiversity hotspot in China. However, fish biodiversity has declined rapidly in this area as the result of human activities and the consequent environmental changes. Beta diversity was a key concept for understanding the ecosystem function and biodiversity conservation. Beta diversity patterns are evaluated and important information provided for protection and management of fish biodiversity in the Luoxiao Mountains. The results showed that the spatial turnover component was the main contributor to beta diversity of Hemiramphidae, Amblycipitidae, Catostomidae, Clariidae, Balitoridae and Percichthyidae in the Luoxiao Mountains, which indicated that a number of protected areas would be necessary to conserve fish biodiversity and that these families would need conservation measures. Most protected areas are currently limited to some regions; therefore, in order to protect fish diversity, conservation efforts must target an increase in the number of protected areas which should be spread across each of the regions.