Based on the speciality of the corresponded relationship between three-dimensional fluorescence spectrum and water bodies or pollution discharge sources, which were relied on the variation of the species and concentrations of fluorescent organic matters, the technique of aqueous fluorescence fingerprint can be applied in pollution discharge identification by the three-dimensional fluorescent characteristic of the water body. In this study, a case was carried out mainlyin River A of City C, southern China, in which the technique of aqueous fluorescence fingerprint was applied to identify the characteristics of the aqueous fluorescence finger print and pollution discharge. The aqueous fluorescence fingerprint of River A showed a prominent characteristic of textile wastewater pollution with three typical fluorescent peaks on [excitation, emission] wavelength of [280, 320], [235, 345] and [255, 460] nm and the similarities among up-, mid- and down-stream of River A were all over 99%. River J diverts into River A from the source of River A. The similarity between River J and A was lower than 60%, and the fluorescent intensity of River J was not more than 40% of River A, which indicated that River J had little influence on the formation process of the aqueous fluorescent fingerprint of River A, and the fluorescence intensity of River A is mainly contributed by its upstream region. River A's pollution might come from the textile wastewater of the printing and dyeing textile industrial park in upstream area, with the aqueous fluorescence fingerprint similarity index of 94%. The linear correlation indexes between the fluorescent peak intensity and the permanganate index of river water were 0.956 4, 0.937 5 and 0.985 4, respectively, and the aqueous fluorescence fingerprint method showed higher sensitivity in pollution perception. Compared with the results of other three-dimensional fluorescence spectral similarity algorithms, the technique of aqueous fluorescence fingerprint for water quality is a reliable water environment monitoring technology and provided a useful tool for the further achievement of pollution source control and precise environmental supervision.