A new species, belonging to the Hypsiboas polytaenius clade, is described from the Parque Estadual do Cerrado (24°10′S, 49°40′W, 659 m elevation), Municipality of Jaguariaíva, State of Paraná, Southern Brazil. Hypsiboas jaguariaivensis sp. nov. is characterized by its medium size (SVL 23.9–28.8 mm in males), head width about 29% of SVL, snout rounded in dorsal and lateral views, supra-cloacal crest and calcar absent, webbing poorly developed between fingers, moderately developed between toes, and dorsal color pattern composed of two pale brown longitudinal stripes extending from the snout to the end of body with a longitudinal sinuous, thin, dark brown stripe between them at mid-body and three pale brown, thin stripes laterally to them; delimiting the dorsal surface and flanks, a conspicuous white stripe extending from the snout, passing over the outer edge of the upper eyelid, to the end of body, delimited dorsally and ventrally by a dark brown line; laterally, a wide, dark brown stripe extending from the snout to the groin, passing over the eye and tympanum; below this wide stripe, a white stripe from the tip of snout, passing over the maxillary border, below the tympanum, and reaching the middle of the flank. The dorsal color pattern separates the new species from all others of the H. polytaenius clade.