Three-dimensional excitation-emission matrix fluorescence spectroscopy coupled with parallel factor analysis was adopted to investigate the characteristics of dissolved organic matter (DOM) components in water samples collected from the Tuojiang River Basin in Chengdu, including its main stream and tributaries. Four DOM components that matched with three fluorescence peaks were identified in the whole river basin and tributaries; while three components corresponding to four fluorescence peaks were identified in the main stream. In all cases, humic-like components accounted for high proportions of the DOM. Correlation analysis revealed the same sources for four components in the whole river basin and its tributaries, whereas two components had different sources in the main stream. Ultraviolet absorbance parameters (SUVA254, SR) and fluorescence parameters (BIX, HIX, FI, β:α) indicated the dominant autochthonous sources of DOM in the whole river basin. Higher terrestrial inputs of DOM were observed in the tributaries than in the main stream. In the areas influenced by human activities (6#, 17#, 18#), the sources of DOM showed strong terrestrial characteristics and high degrees of humification and aromatization, as well as serious pollution. The results of this study have potentially far-reaching implications for environmental water management in the area.