The widespread occurrence of microplastics (MPs) has resulted in their interaction with biological processes. Thus, there is a great concern about the potential toxicity of MPs on animal and plant cells and on the possibility that MPs reach humans through the food web. In order to shed light on both issues, laboratory assays were performed for evaluating the effects of polyethylene (PE) microparticles on the aquatic plant Spirodela polyrhiza (L.) Schleid. and the gammarid Echinogammarus veneris (Heller, 1865). Moreover, a stock of MP-treated Spirodela plants was used to feed gammarid individuals, and the presence of MP particles in their digestive tracts was analyzed. Results evidenced the lack of toxic effects of MPs on plants, evaluated at growth and physiological level by biometric parameters, pigment content, and photosynthetic performance estimated by chlorophyll fluorescence imaging through the ETPT (EcoTox Photosystem Tool). Only a slight reduction in pigment-related indices in MP-treated plants was observed. A remarkable genotoxic effect was instead highlighted by Comet assay in the hemocytes of gammarid individuals exposed to MPs, with three times more DNA damage (expressed as Tail Moment) in MP-treated individuals compared to control ones. Finally, the gut content of the gammarids fed with MP-treated plants revealed the presence of 7.6 MP particles/individual, highlighting the occurrence of trophic transfer of MPs among freshwater ecosystem organisms. Novel indications about the potential impact of the PE microparticles in the aquatic compartment are provided. Notably, the transfer of MP particles between primary producer and primary consumer organisms of the freshwater trophic chain and the genotoxic effects associated with the ingestion of such particles by gammarids are issues of concern for the aquatic ecosystem and the food web leading to the human diet.