Abstract: The important influence of the planning, construction and opening of the High-Speed Railway on the development of urban tourism is to shorten the time distance of spatial connection of urban tourism, and to enhance the spatial accessibility of urban tourism traffic, and to improve further the industrial connection between the central and external cities and the intensity of spatial flow of elements. Taking Guiyang-Guangzhou High-Speed Railway(GGHSR) as a typical area, this paper uses the accessibility model and GIS spatial analysis to measure the influence of High-Speed Railway on temporal distance of tourism cities and construct the regional structural system.The results show that: (1) The shortest time distance of the tourism cities along the GGHSR is obviously shortened after the opening of the GGHSR. (2) The changing rate of accessibility of Congjiang, Sanjiang, Duyun, Guilin, Zhaoqing, Huaiji, Yangshuo, Guiyang, Guangzhou shows decreasing trend after the opening of the GGHSR. (3) The Grade III Guizhou tourism development plate and the Grade III Guangxi tourism development plate and the Grade II Guangdong tourism development plate and the Grade I Guiyang-Guangzhou High-Speed Railway tourism development plate along the GGHSR are formed based on the certainty of different degrade tourism development nodes and tourism development axises along the GGHSR, and the regional structure system of tourism cities along the GGHSR is constructed further. The above conclusions provide scientific foundation for enhancing reasonably the spatial structure of tourism cities along the GGHSR and coordinating the integrated tourism development along the GGHSR.