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Investigating fish diversity in canyon rivers through conventional fish surveys is challenging due to precipitous conditions, including steep slopes, rapid water flow, and complex habitats. Additionally, intensive construction of dams has further complicated the understanding of contemporary fish diversity in these rivers. In this study, we used the environmental DNA (eDNA) technique to assess fish diversity and examine the effects of dams on fish diversity in the Mabiehe River, a canyon river in the upper reaches of the Pearl River drainage. Water samples from 15 sampling sites were collected, yielding 9,356,148 valid sequences. Utilizing the NCBI public database, a total of 60 freshwater fish species were identified, with <i>Carassius auratus</i>, <i>Cyprinus carpio</i>, and <i>Pelteobagrus fulvidraco</i> being the most dominant species in the Mabiehe River. We also detected one nationally protected fish species, three provincially protected fish species, and six exotic species in this river. Furthermore, eDNA analyses demonstrated that the lotic river sections harbor more species and greater diversity than dammed sections, suggesting that dams might exert significant impacts on local fish diversity. Overall, this study supports the effectiveness of the eDNA technique as a complementary tool to traditional field surveys for monitoring fish biodiversity in canyon rivers.