Dissolved organic matter (DOM) and nutrients have a unique way of producing, decomposing and storing in southwest karst water systems. To understand the biogeochemical cycle of DOM in karst aquifer systems, we investigated the behavioral changes of DOM fluorescence components in Zhaidi karst river system. Two humic-like components (C1 and C2), and one autochthonous tyrosine-like component (C4) were identified using the parallel factor analysis (PARAFAC) model. Compared with the traditional physical and chemical indicators, spatial heterogeneity of DOM was more obvious, which can reflect the subtle changes in groundwater system. Traditional indicators mainly reflect the regional characteristics of karst river system, while DOM fluorescence components reflect the attribute gaps of sampling types.