Hypnea musciformis, a red macroalga widely distributed in tropical and subtropical coasts around the world, has great economic importance as a source of carrageenan for industrial production. In this work, the DNA barcode marker COI-5P and the plastid rbcL gene, in addition to morphological studies, were used to investigate this species on the coast of Brazil and compare it with specimens from other countries. A total of 128 sequences were obtained in this study for 100 specimens from Brazil and 15 specimens from other countries, including the type locality in Italy. The divergence between South American sequences and sequences from Italy for H. musciformis was significantly high for both markers, indicating that the specimens found on the Brazilian coast belong to a different species. Considering the data gathered from molecular markers and morphological analysis, the specimens previously identified morphologically as "H. musciformis", "Hypnea nigrescens", and "Hypnea valentiae" collected in Brazil were considered morphological variations of the new species described in this paper, named Hypnea pseudomusciformis Nauer, Cassano & M.C. Oliveira, sp. nov. The identification of specimens based only on morphological characteristics proved to be unsatisfactory for reasons that could be attributed to phenotypic plasticity in this species. Thus, the technique of DNA barcoding, especially with respect to the COI-5P marker, was essential for the identification and definition of species, revealing scenarios that would otherwise be ignored by using only morphological analysis.