Long-term presence of waste tires in the soil brings negative effects and risks for the environment. Waste tires can be found in the open nature long after they have been left there. Their chemical composition and components are degradable and contaminate the environment. The research is focused on the evaluation of chemical changes in soils exposed to the long-term action (disposal) of waste tires and on the determination of the degree of phytotoxicity of such soils. Soil samples were collected from 4 sites in the territory of the Czech Republic. The samples were subjected to chemical analyses in an accredited laboratory. The analyses revealed markedly exceeded limits for heavy metals, namely Pb, Zn and Hg in some soil samples, as compared with the Decree No. 153/2016 Coll. Ranges of exceeded limits were from 40% to 26 566%. Then the samples were evaluated by means of microbiotests (acute phytotoxicity by means of Phytotoxkit test set, 72 hours) and also by means of long-term phytotoxicity test using the subchronic container test (28 days). The tests of phytotoxicity were made with the seeds of Sinapis alba L. and Lepidium sativum L. Results of subchronic container tests showed the percentage of germination (PG) in the soil samples ranging from 74.36%-90.48%. A sample with the PG value lower than 90% was evaluated as phytotoxic compared with the control substrate (100%). In the acute microbiotest, the soil samples exhibited values of root growth inhibition (IR) ranging from 5.12%-50.98%. Substrates with the IR value (%) over 0 showed inhibition/retardation of root growth in the tested plant, and the soil is considered to be phytotoxic.