Floral morphological traits are frequently used to identify species, including those that are closely related and show low genetic diversity, and floral shape and colour are known to play an important role in diversification and species isolation. Floral morphology in Petunia (Solanaceae) is considered a driver of diversification because of its association with pollinators. Here, flower morphology was characterized through morphometric analyses and floral pigments. Our main aim was to determine corolla shape in populations of Petunia axillaris and P. exserta and their natural hybrids and how floral display, size and colour are involved in pollinator attraction. In addition, we investigated floral pigments in P. exserta and different hybrid classes. The results from morphometric analyses revealed that each species has a specific floral shape, independent of the collection site. By contrast, in two contact zones, a mosaic of floral phenotypes was observed with some hybrid classes based on corolla colour being placed close to P. exserta. The results suggest that several generations of hybrids or backcrossing could have given rise to this floral diversity in contact zones.

Population genetic structure results from the interaction between historical events, current ecological conditions and life traits. The genetic structure and gene flow between populations are important to species dynamics, mainly for rare and endangered species that are more vulnerable to landscape changes and fragmentation. Here we evaluated the genetic diversity, population structure and gene exchange in Petunia bonjardinensis, P. reitzii and P. saxicola, three rare species endemic to subtropical highland grasslands in southern South America. We analysed the genetic diversity and structure considering historical events, such as founder effect and climate changes, and biological traits of each species. We also estimated the conservation status for these three species. We collected samples from all adult individuals and occurrence sites that could be found at the same flowering season and genotyped them for 13 nuclear microsatellite markers. Our results indicate that rarity is probably historical for these species, given that we found no genetic evidence for recent bottlenecks. Petunia bonjardinensis, with the largest occurrence area and population sizes, displayed the higher diversity indices. The other two showed lower genetic diversity and are geographically most restricted. Gene exchange among these species was low, although they share some ancestral genetic polymorphism. Historical migration, founder effects and Pleistocene climate cycles ae the main factors explaining genetic diversity, and this was also influenced by reproductive biology and recent habitat loss, whereas the landscape influences the structure. Based on IUCN criteria, the three species are endangered, and the main risk for their survival is probably anthropic activity in the occurrence area. We recommend an urgent programme for the preservation of these species in situ and ex situ.

Interspecific hybridization has been fundamental in plant evolution. Nevertheless, the fate of hybrid zones throughout the generations remains poorly addressed. We analyzed a pair of recently diverged, interfertile, and sympatric Petunia species to ask what fate the interspecific hybrid population has met over time. We analyzed the genetic diversity in two generations from two contact sites and evaluated the effect of introgression. To do this, we collected all adult plants from the contact zones, including canonicals and intermediary colored individuals, and compared them with purebred representatives of both species based on seven highly informative microsatellite loci. We compared the genetic diversity observed in the contact zones with what is seen in isolated populations of each species, considering two generations of these annual species. Our results have confirmed the genetic differentiation between the species and the hybrid origin of the majority of the intermediary colored individuals. We also observed a differentiation related to genetic variability and inbreeding levels among the populations. Over time, there were no significant differences per site related to genetic diversity or phenotype composition. We found two stable populations kept by high inbreeding and backcross rates that influence the genetic diversity of their parental species through introgression.

The Brazilian Atlantic Rainforest is one of the most diverse and threatened ecoregions on the planet and displays high levels of endemism. Despite several population analyses and phylogeographical studies, the origins of its species richness and the evolutionary processes that gave rise to this diversification remain poorly understood, especially at the southern edge of the Atlantic Forest. Passiflora actinia is an indigenous species from the southern Atlantic Forest and, as such, was influenced by climatic changes during the Pleistocene. In this study, we investigated the effects of past climate changes on the genetic diversity of P. actinia, using nuclear and plastid markers. We subsequently suggest strategies for the preservation of this species in particular and the whole ecoregion in general. We employed phylogeographical methods and combined these results with past, present and future ensemble niche models. Genetic variability in P. actinia was similar to that of other species with similar geographical distributions, and geographical structuring was not observed based on either type of genetic marker. Diversification in P. actinia was dated to the Pleistocene, suggesting that climate changes could have influenced the distribution of genetic diversity in this species. Our results predicted that suitable P. actinia habitat will persist in the highlands but will be reduced in the lowlands, especially with higher greenhouse gas concentrations. Conservation efforts should focus on populations with unique genetic units and populations from areas with greater climatic instability. Habitat loss due to deforestation in the Atlantic Forest constitutes a major risk to this species, especially to small populations or those with low diversity indices.

Petunia axillaris subsp. axillaris and P. axillaris subsp. parodii (Solanaceae) are closely related taxa primarily distinguished by their geographical distribution and floral morphology. Both are found in the southern South American Pampas ecoregion, with the latter also occurring in the Chaco ecoregion; in the Pampas, taxa occupying adjacent areas and intermediary morphologies have been observed. Here, we explored morphological traits and genetic polymorphisms to test the occurrence of hybrid zones between these taxa along the Negro River (Uruguay) and southern Brazil. Morphological data showed variation among the groups, although they could have some overlap. Genetic data also support the differences between them and hybrid zones. These findings suggest that the divergence between P. axillaris subspecies occurred with occasional hybridization in their contact zones.