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Four species of Pluteus Fr. are documented based on recently collected specimens from Alto Paraguay and Central Department, which correspond to the Pantanal and Humid Chaco ecoregions, respectively. Pluteus fibulatus Singer, P. longistriatus (Peck) Peck, P. petasatus (Fr.) Gillet, and P. triplocystis Singer are newly recorded species of the mycobiota of Paraguay; P. triplocystis is also a newly recorded for South America. Descriptions and photographs of both macro- and microscopic features of each species are included along with a brief discussion on their taxonomy, ecology, and distribution.