| Summary: | The maize pathogen <i>Fusarium verticillioides</i> and their mycotoxins cause damage to plants, animals, and human health. This work aimed to evaluate the effect of crude extracts (CEs) from <i>Agaricus subrufescens</i>, <i>Lentinula edodes</i>, and <i>Pleurotus ostreatus</i> fruiting bodies on in vitro production of biomass and mycotoxins by two strains of <i>F. verticillioides</i>. Stipes and pilei were separated before extraction for <i>A. subrufescens</i> and <i>L. edodes</i>. Comparative metabolomics and dereplication of phenolic compounds were used to analyze all CEs. Mushroom CEs did not significantly inhibit the production of mycelial biomass at concentrations of 2 mg mL<sup>−1</sup>. CEs from <i>A. subrufescens</i> (stipes and pilei) and <i>L. edodes</i> pilei inhibited the production of fumonisins B1 + B2 + B3 by 54% to 80%, whereas CE from <i>P. ostreatus</i> had no effect. In contrast, CE from <i>L. edodes</i> stipes dramatically increased the concentration of fumonisins in culture media. Fusaric acid concentration was decreased in cultures by all CEs except <i>L. edodes</i> stipes. Differences in phenolic composition of the extracts may explain the different effects of the CE treatments on the production of mycotoxins. The opposing activities of stipes and pilei from <i>L. edodes</i> offer an opportunity to search for active compounds to control the mycotoxin production by <i>F. verticillioides</i>.
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