A comparison of different transportation route types and their combined effects on landscape diversity was conducted within Tiaoxi watershed (China) between 1994 and 2005. Buffer analysis and Mann-Kendall's test were used to quantify the relationships between distance from transportation routes (railway, highway, national, and provincial road) and a family of landscape diversity parameters (Simpson's diversity index, Simpson's evenness index, Shannon's diversity index, and Shannon's evenness index). One-way ANOVA was further applied to compare influences from different route types and their combined effects. Five other landscape metrics (patch density, edge density, area-weighted mean shape index, connectance index, and Euclidean nearest neighbor distance) were also calculated to analyze the associations between landscape diversity and landscape pattern characteristics. Results showed that transportation routes exerted significant impacts on landscape diversity. Impact from railway was comparable to that from highway and national road but was more significant than that from provincial road. The spatial influential range of railway and national road was wider than that of highway and provincial road. Combined effects of routes were nonlinear, and impacts from different route types were more complex than those from the same type. The four landscape diversity metrics were comparably effective at the buffer zone scale. In addition, landscape diversity can be alternatively used to indicate fragmentation, connectivity, and isolation at route buffer scale. This study demonstrates an applicable approach to quantitatively characterize the impacts from transportation routes on landscape patterns and has potential to facilitate route network planning.