We investigated Lepiota species from Benin and reference species from Europe based on fieldwork, light microscopy, and the analysis of DNA sequence data. As a result, thirteen Lepiota species from Benin are described as new to science, whereas only one previously described species, L. albogranulosa, was found. An identification key to these species is provided. The taxonomical results are supported by a comprehensive multilocus phylogeny based on ITS, LSU, and RPB2 sequences. In addition, the occurrence of amatoxins in basidiomata of Lepiota spp. was examined, in order to detect toxic mushrooms and for chemotaxonomic conclusions for the genus Lepiota in general. To this end, an UHPLC-HRMS2 analysis process was applied and molecular networks were generated to obtain an overview of the chemical diversity encompassed by 46 Lepiota species, with particular emphasis on their amatoxin content, resulting in the first report of a Lepiota species containing amatoxin for Benin. The annotation coverage was extended using DEREPLICATOR and taxonomically informed metabolite annotation (TIMA). Birnbaumins were discovered in several species as well as other chemical features of possible chemotaxonomic interest. This study extends the knowledge about diversity, amatoxin occurrence, and possible chemotaxonomic markers in the genus Lepiota. Citation: Sarawi S, Reschke K, Jagora A, Yorou NS, Le Pogam P, Piepenbring M (2025). Hidden gems of Benin: Unravelling the diversity of Lepiota spp. through phylogenetic, morphological, and chemotaxonomic exploration. Persoonia 55: 313-360. doi: 10.3114/persoonia.2025.55.09.
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