[Objective] The ecological vulnerability and spatial variation characteristics of the Wujiang River basin in Guizhou Province were studied to provide a scientific reference for the ecological environment construction and protection in the area.[Methods] By constructing the evaluation index system, the AHP was used to weight the ecological vulnerability factors of the basin, and GIS was used to comprehensively evaluate the ecological vulnerability of the basin.[Results] ① The three classifications of quantile, equal interval and natural discontinuity had little effect on the classification of ecological vulnerability, but there were significant differences in area ratio, and the proportions of extremely fragile and potentially vulnerable watershed were the smallest. ② The ecological fragility of the basin is mainly fragile and moderately fragile, the areas of the two fragilities accounting for more than 50%. ③ Human activities have a significant impact on the ecological vulnerability of the basin. The surrounding areas of large and medium-sized cities such as Guiyang and Zunyi City are mostly and extremely vulnerable. ④ The spatial distribution of the generally fragile areas in the basin is relatively scattered, which is not conducive to the ecological environment protection of the basin.[Conclusion] The ecological vulnerability level of the basin is gradually reduced from the upstream to the downstream, and the spatial distribution characteristics of vulnerability are obvious. The protection of the ecologically fragile areas of the basin in terms of the local conditions has a great effect on reducing the ecological vulnerability of the basin.