Salinization in freshwater lakes is becoming a serious global environmental problem, especially in lakes of plateaus such as south-western plateau of China. However, limited information is available about the molecular response of freshwater hydrophytes to salinity under multiple stress. In the present study, a weighted gene co-expression network (WGCNA) was used to identify the modules of co-expressed genes in the physiological and biochemical indicators of Pistia stratiotes to determine its molecular response to salinity (NaCl) alone and when combined with cadmium (Cd). The physiological and biochemical indicators showed that P. stratiotes improved its salt tolerance by enhancing photosynthetic abilities, reducing oxidative stress, and inducing osmoprotectant generation. Morever, addition of NaCl reduced the Cd accumulation in P. stratiotes. Transcriptome and WGCNA analysis revealed that the pathways of alpha-linolenic acid metabolism, ribosomal, flavonoid biosynthesis, and phenylpropanoid biosynthesis were significantly enriched in both treatments. Genes associated with photosynthesis-antenna proteins, nitrogen metabolism, and the acid cycle pathways were only expressed under salinity stress alone, while the proteasome pathway was only significantly enriched in the combined salinity and Cd treatment. Our findings provide novel insights into the effects of salinization on aquatic plants in freshwater ecosystems and the management of aquatic ecosystems under global change.