This study provides quantitative evidence on the spatial distribution of rural tourism resources and their relationship with transportation accessibility in Northern Anhui Province, China. Using spatial analysis of tourism development patterns across five core counties, the results reveal a strong association between transportation infrastructure density and the concentration of tourism resources. Counties with higher densities of roads, expressways, and railway stations exhibit significantly greater clustering of tourism attractions. In contrast, larger administrative units such as Fuyang and Suzhou, despite possessing notable scenic resources, demonstrate weaker tourism development trends, largely due to insufficient transportation accessibility and dispersed resource distribution. The findings indicate that administrative scale and urban–rural land allocation structures play a decisive role in shaping tourism spatial patterns, with smaller counties enjoying a natural advantage in achieving resource concentration. Conversely, tourism resources in larger counties tend to be spatially fragmented, which constrains tourism growth and related economic activities. Furthermore, ordinary least squares (OLS) regression analysis confirms that transportation accessibility is the most influential factor affecting the spatial agglomeration of rural tourism resources, explaining approximately 69% of the observed variation. Better accessibility not only enhances connectivity and reduces travel time but also promotes investment in tourism infrastructure and services. These findings offer important policy implications for optimising transportation planning and promoting balanced rural tourism development in Northern Anhui Province.