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There is an ongoing interest in the production of mushrooms as food, and with their consumption on the rise, there is a need to establish different safety measures. In the present study, six mushroom species (Agrocybe cylindracea, Clitocybe maxima, Flammulina velutipes, Ganoderma lucidum, Lentinula edodes and Pleurotus eryngii) were cultivated on two commonly used substrates (A—based on alder and beech sawdust; B—based on oak sawdust and flax shives). The aims were to determine their growth and the accumulation of 70 elements in their fruiting bodies, 33 of which were detected in all analysed samples and were used for statistical evaluation. Cultivation of C. maxima, G. lucidum and L. edodes resulted in higher yield and mineral content when substrate A was used, cultivation of A. cylindracea and F. velutipes yielded better results on substrate B, P. eryngii cultivation outcomes were similar for both substrates. L. edodes was found to have a high affinity to accumulate Cd, C. maxima can bioconcentrate Al and Ni, Ganoderma lucidum—Pb, while F. velutipes—Hg. The study indicated that the chemical composition of substrates could affect both the yield and the level of various toxic and nutritional elements. [ABSTRACT FROM AUTHOR]