Silver nanoparticles (AgNPs), a component of many consumer products, are considered an environmental risk due to the broad-spectrum toxicity of Ag⁺ to non-target organisms. Most AgNPs released from consumer products will end up as biosolids in wastewater treatment plants, which are often applied as a fertilizer to agriculture. Land application of biosolids may add AgNPs to the soil–plant system, with unknown consequences. This study investigated the growth of Hordeum vulgare seedlings, Ag bioconcentration and distribution in shoot and root tissues of barley exposed to biosolid-amended Delacour and Organization for Economic Co-operation and Development (OECD) soils spiked with AgNPs (up to 366 mg Ag kg^-1 dry soil). In both soils, root and shoot growth declined linearly as the concentration of AgNPs increased. Barley had higher Ag bioconcentration values when grown in the OECD soil than in the Delacour soil. Silver bioavailability was greater in the OECD soil due to its physicochemical properties, such as low calcium concentration and acidic pH, relative to the Delacour soil. Barley seedlings exhibited morphological changes, including smaller shoots and shorter, thick roots after 14 d exposure to AgNPs. We conclude that plant structural responses, particularly changes in root biomass, could be an early diagnostic of seedling exposure to AgNPs in biosolid-amended soils.