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This paper presents the floristic survey of ferns and lycophytes occurring in a gallery forest in the central region of Mato Grosso do Sul, Brazil. In the study area, 29 species and 2 varieties were recorded. Dryopteridaceae and Pteridaceae were the richest families (8 and 5 species, respectively) and Elaphoglossum and Blechnum were the richest genera (3 species each one). Preferably, the listed species occur within the forest (68%), they occupy the terrestrial substrate (77.4%), and they are hemicryptophyte (77.4%) and rosulate (64.5%). We observed four species still not mentioned for Mato Grosso do Sul, which are Blechnum lanceola L., Elaphoglossum pachydermum (Fée) T. Moore, Lindsaea lancea (L.) Bedd var lancea, and Mickelia nicotianifolia (Sw.) R. C. Moran et al., which has its southern limit of distribution in Brazil, in the study area.