Abstract

Abstract Closely related and young species often show high morphological similarity, challenging their identification and correct assignment. Molecular markers and integrative approaches have contributed to solving many taxonomic uncertainties. In this study, we evaluated the genetic variability and ecological features of Petunia guarapuavensis and Petunia scheideana, over which there is a taxonomic debate. Both species are endemic and rare, distributed in the subtropical highland grasslands in southern South America. We based our analyses on nuclear microsatellite and plastid sequences, aiming to disentangle the taxonomic ambiguities that made some consider these entities synonymous despite occupying different clades in the genus phylogenetic tree. Our findings support that there is genetic differentiation between these species, suggesting that they are independent taxonomic entities despite sharing floral traits and a few molecular polymorphisms. The low genetic sharing between the species is likely due to a common ancestor and recent divergence time. In contrast, their morphological similarity can be attributed to the absence of selective pressure, as both grow under similar ecological conditions. This study emphasizes that adding more than one sequence per species, combining data with dissimilar inheritance patterns, and exploring data through different methodologies help to disentangle taxonomic incongruences and reveal diversity that might otherwise remain hidden.

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