To understand the spatial species diversity of demersal fish assemblages in Taijiang National Park (TJNP) of Taiwan, fishes from 44 demersal trawl hauls and environmental data were collected in the nearshore and offshore areas of TJNP from April 2016 to May 2019. In total, fishes of 47 families, 84 genera, and 113 species were recorded. The nearshore and offshore demersal fish assemblages in TJNP exhibited significant variability in species composition assessed via beta diversity. Using distance-based redundancy analysis, we demonstrated that bottom depth and substrate type were significant explanatory variables of spatial species diversity and identified three habitat types (I: shallow soft bottom; II: deeper soft bottom; III: deeper bottom with mixed sand and gravel substrates). The nearshore assemblage was characterized by type I, where Tarphops oligolepis (flounder), Trachinocephalus myops (snakefish), and Liachirus melanospilos (carpet sole) dominated in terms of abundance. The offshore assemblage was characterized by either type II or type III because differences in substrate types among sampling sites were noticeable. At the offshore sites characterized by a deeper soft bottom (type II), Johnius distinctus (croaker), Cynoglossus kopsii (shortheaded tonguesole), and Coelorinchus formosanus (Formosa grenadier) predominated. In contrast, the westernmost sampling site, characterized by type III habitat, exhibited relatively high Shannon indices, and Scorpaena miostoma (scorpionfish), Urolophus aurantiacus (sepia stingray), and Parabothus taiwanensis (lefteye flounder) predominated. Our results provide the first baseline information on the environmental characteristics and spatial species diversity of demersal fish assemblages in TJNP and have implications for biodiversity conservation in existing spatial management areas.