Identifying discontinuous entities within species complexes is a major topic in systematic and evolutionary biology. Comprehensive inventories describing and identifying species rapidly and correctly before they or their habitats disappear is especially important in megadiverse regions, such as South America continent, where a large part of the biodiversity is still unknown and remains to be discovered. Species complexes may account for a substantial number of plant groups in the South American flora, and studies investigating species boundaries in such challenging groups are needed. In this context, multidisciplinary approaches are crucial to understanding the species integrity and boundaries within species complexes. Morphometrics, cytogenetics, anatomy, crossing experiments, and molecular markers have been combined in different ways to investigate species complexes and have helped depict the mechanisms underlying the origin of South American species. Here, we review the current knowledge about plant species complexes on the hyperdiverse South American continent based on a detailed examination of the relevant literature. We discuss the main findings in light of the potential evolutionary mechanisms involved in speciation and suggest future directions in terms of integrating multispecies coalescence methods with several complementary types of morphological, ecological, and geographical data in this research field.