The Shennongjia Forestry District (Shennongjia) located in the northwestern region of Hubei Province, serves as the watershed between the upper and middle Chang-Jiang basin. Due to the unique natural environment and well-developed river networks, Shennongjia harbors a relatively high species diversity of freshwater fish, thus making it one of the hotspots areas of biodiversity research in China. However, there is still a lack of essential understanding of the fish diversity in this region. An updated checklist of fishes in the Shennongjia is provided, based on the results of the field survey conducted from 2021 to 2022 and combined with historical records of fish diversity in this district and the latest taxonomic progress of relevant taxa in the known literature.This updated checklist includes a total of 52 species, 44 of which are native and eight alien. The Cypriniformes dominates the fish fauna of Shennongjia. Two families of Cyprinidae and Xenocyprididae have the highest number of species. There are six newly recorded species, three previously undescribed species, and 12 species undergoing nomenclatural changes. Eight historically recorded species are excluded from this list, but 27 historically recorded species uncollected during this field survey are remained.The updated checklist presented here can serve as a fundamental reference for future biodiversity analyses and the biodiversity conservation of Shennongjia.