DNA barcoding is a technique for discriminating and identifying species using short, variable, and standardized DNA regions. Here, we tested for the first time the performance of plastid and nuclear regions as DNA barcodes in Passiflora. This genus is a largely variable, with more than 900 species of high ecological, commercial, and ornamental importance. We analyzed 1034 accessions of 222 species representing the four subgenera of Passiflora and evaluated the effectiveness of five plastid regions and three nuclear datasets currently employed as DNA barcodes in plants using barcoding gap, applied similarity-, and tree-based methods. The plastid regions were able to identify less than 45% of species, whereas the nuclear datasets were efficient for more than 50% using “best match” and “best close match” methods of TaxonDNA software. All subgenera presented higher interspecific pairwise distances and did not fully overlap with the intraspecific distance, and similarity-based methods showed better results than tree-based methods. The nuclear ribosomal internal transcribed spacer 1 (ITS1) region presented a higher discrimination power than the other datasets and also showed other desirable characteristics as a DNA barcode for this genus. Therefore, we suggest that this region should be used as a starting point to identify Passiflora species.

Hybridization between closely related plant species is a widespread phenomenon with significant evolutionary consequences, so natural hybrid zones provide exciting opportunities to study the processes of genetic differentiation and species formation. In Petunia, genetics, ecology and evolution of pollination syndromes have been thoroughly studied and related to the development of prezygotic isolation barriers between species. However, to date, no studies have explored the importance of extrinsic post-zygotic barriers such as environmental selection as drivers of reproductive isolation. In this study, we applied a population genetics approach to understand the speciation of two partially sympatric taxa of Petunia Juss that hybridize in the wild despite having strong differences in pollination syndrome, mating system and environmental requirements. We evaluated 58 individuals from seven allopatric and two contact zones of Petunia axillaris and P. exserta with genomic data to study the population genetic structure and gene exchange between these species and scan for potential loci under natural selection related to the preservation of species barriers. We explicitly evaluated the time, intensity and direction of interspecific migration, testing for alternative demographic scenarios. The genetic structure showed well-differentiated species-level lineages, and poor differentiation among populations of P. axillaris; P. exserta populations were highly structured, with several well-differentiated intraspecific lineages. Our data provide evidence for admixture with low asymmetric gene flow from P. axillaris to P. exserta associated with recent secondary contact, suggesting that pollinator specificity is not the only trait responsible for species integrity in this system. Combining the results from FST outlier and genotype–phenotype association methods, we identified 54 non-synonymous candidate variants under natural selection. Those variants are found in 35 coding sequences from which we found genes related to light-response networks, supporting that extrinsic factors such as habitat preferences can also provide a mechanism of reproductive isolation between Petunia spp. We argue that a set of pre- and post-zygotic barriers in conjunction with demographic processes maintain the species integrity.

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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.

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.