História evolutiva das espécies de Passiflora L. de ocorrência no Rio Grande do Sul: aspectos genéticos, estrutura populacional e filogenia molecular

Passiflora L. is the largest genus in Passifloraceae family, presents preferentially Neotropical distribution and nowadays is split in four subgenera. Nineteen species are described to Rio Grande do Sul that are deeply studied by our research group about evolutionary relationships and genetic aspects. Here we present the main results on phylogeny and population structure.O gênero Passiflora L. é o maior dentro da família Passifloraceae. Apresenta distribuição predominantemente Neotropical e está atualmente dividido em quatro subgêneros. Para o Rio Grande do Sul, são relacionadas 19 espécies, que têm sido amplamente estudadas do ponto de vista evolutivo e genético por nosso grupo de pesquisas. Aqui são apresentados os principais resultados sobre filogenia e estrutura populacional

Phylogenetic position of monotypic genus Verbenoxylum (Verbenaceae) and new combination under Recordia

In spite of the recent studies on the phylogeny of Verbenaceae, the position of the monotypic Verbenoxylum, endemic to the Atlantic rainforest in southeastern Brazil, remains unsolved. Molecular data were here analyzed to infer the phylogenetic placement of this genus; furthermore morphological data was studied in order to examine traits that support relationships among taxa. Sequences of the plastid regions of ndhF gene and trnL—trnF intergenic spacer were analyzed to conduct phylogenetic studies with maximum parsimony, maximum likelihood, and Bayesian inference. Morphological traits that had been traditionally used to distinguish tribes within Verbenaceae, as well as those employed to characterize Verbenoxylum, were examined. Verbenoxylum is nested within the tribe Duranteae, sister to Recordia, a monotypic genus endemic to Bolivia, a placement never reported before. The morphological traits analyzed prove not to be useful to distinguish tribes but are important at lower taxonomic levels. Based on the sister relationship and morphological similarities between the genera Verbenoxylum and Recordia, we propose the inclusion of Verbenoxylum reitzii into Recordia, forming the new combination Recordia reitzii.Fil: Thode, Veronica. Universidade Federal do Rio Grande do Sul; BrasilFil: O'leary, Nataly Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica Darwinion. Academia Nacional de Ciencias Exactas, Físicas y Naturales. Instituto de Botánica Darwinion; ArgentinaFil: Olmstead, Richard. University of Washington; Estados UnidosFil: Freitas, Loreta. Universidade Federal do Rio Grande do Sul; Brasi

Species boundary and extensive hybridization and introgression in Petunia

Studying the role of hybridization in the speciation of plants is one of the most thrilling areas of evolutionary biology. Hybridization in natural populations can act in opposition to divergence, contribute to adaptation through introgression or foster the emergence of new lineages via hybrid speciation. Species of the plant genus Petunia grow in open areas in southern South America. Some natural interspecific hybrid events have been described for the genus, such as between the endemic P. exserta and the widespread P. axillaris. Both species occur in sympatry in Serra do Sudeste (Brazil), where they occur in diverse habitats and exhibit floral divergence, which has been related to the attraction of different primary pollinators. The present study evaluates the maintenance of the species boundaries front of hybridization and introgression. Direct and indirect methods of estimating gene exchange employed genotyping 720 reproductive plants and 611 progenies of both species with eight microsatellite loci. Gene exchange was found to be frequent and bidirectional between the species, indicating that introgression changes their genetic constitution in areas of sympatry. Limits of the studied species are being maintained because of the high level of inbreeding and backcrosses that are habitat-dependent

Phylogenetic analyses of some key genes provide information on pollinator attraction in Solanaceae

Floral syndromes are known by the conserved morphological traits in flowers associated with pollinator attraction, such as corolla shape and color, aroma emission and composition, and rewards, especially the nectar volume and sugar concentration. Here, we employed a phylogenetic approach to investigate sequences of genes enrolled in the biosynthetic pathways responsible for some phenotypes that are attractive to pollinators in Solanaceae genomes. We included genes involved in visible color, UV-light response, scent emission, and nectar production to test the hypothesis that these essential genes have evolved by convergence under pollinator selection. Our results refuted this hypothesis as all four studied genes recovered the species’ phylogenetic relationships, even though some sites were positively selected. We found differences in protein motifs among genera in Solanaceae that were not necessarily associated with the same floral syndrome. Although it has had a crucial role in plant diversification, the plant–pollinator interaction is complex and still needs further investigation, with genes evolving not only under the influence of pollinators, but by the sum of several evolutionary forces along the speciation process in Solanaceae

Molecular evolution and structural analyses of proteins involved in metabolic pathways of volatile organic compounds in Petunia hybrida (Solanaceae)

The association between plants and their pollinators is essential for increasing the diversity in angiosperms. Morphological and physiological traits, mainly floral scent, can influence the pollination dynamics and select pollinators for each plant species. In this work, we studied two proteins involved in producing volatile organic compounds in plants, conyferyl alcohol acyltransferase (CFAT) and benzoyl-CoA:benzyl alcohol/phenyl ethanol benzoyl transferase (BPBT) genes. We aimed to understand these proteins with respect to evolutionary and structural aspects and functions in Solanaceae using phylogenetic methods and comparative molecular modeling. We used Bayesian inference to describe the proteins’ evolutionary history using Petunia x hybrida as a query to search for homologs in the Solanaceae family. Theoretical 3D models were obtained for both proteins using Panicum virgatum as a template. The phylogenetic tree included several different enzymes with diverse biological roles in Solanaceae, displaying the transferase domain. We identified only one sequence of CFAT in the databases, which belongs to Petunia x hybrida, and found several BPBT sequences from the genera Nicotiana, Solanum, and Capsicum. The 3D structures of CFAT and BPBT have two different domains, and we have identified the amino acid residues essential for the enzymatic activity and interaction with substrates

What could be the fate of secondary contact zones between closely related plant species?

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

Single gene mutation in a plant MYB transcription factor causes a major shift in pollinator preference.

Understanding the molecular basis of reproductive isolation and speciation is a key goal of evolutionary genetics. In the South American genus Petunia, the R2R3-MYB transcription factor MYB-FL regulates the biosynthesis of UV-absorbing flavonol pigments, a major determinant of pollinator preference. MYB-FL is highly expressed in the hawkmoth-pollinated P. axillaris, but independent losses of its activity in sister taxa P. secreta and P. exserta led to UV-reflective flowers and associated pollinator shifts in each lineage (bees and hummingbirds, respectively). We created a myb-fl CRISPR mutant in P. axillaris and studied the effect of this single gene on innate pollinator preference. The mutation strongly reduced the expression of the two key flavonol-related biosynthetic genes but only affected the expression of few other genes. The mutant flowers were UV reflective as expected but additionally contained low levels of visible anthocyanin pigments. Hawkmoths strongly preferred the wild-type P. axillaris over the myb-fl mutant, whereas both social and solitary bee preference depended on the level of visible color of the mutants. MYB-FL, with its specific expression pattern, small number of target genes, and key position at the nexus of flavonol and anthocyanin biosynthetic pathways, provides a striking example of evolution by single mutations of large phenotypic effect

Genetic architecture of a pollinator shift and its fate in secondary hybrid zones of two Petunia species.

BACKGROUND Theory suggests that the genetic architecture of traits under divergent natural selection influences how easily reproductive barriers evolve and are maintained between species. Divergently selected traits with a simple genetic architecture (few loci with major phenotypic effects) should facilitate the establishment and maintenance of reproductive isolation between species that are still connected by some gene flow. While empirical support for this idea appears to be mixed, most studies test the influence of trait architectures on reproductive isolation only indirectly. Petunia plant species are, in part, reproductively isolated by their different pollinators. To investigate the genetic causes and consequences of this ecological isolation, we deciphered the genetic architecture of three floral pollination syndrome traits in naturally occurring hybrids between the widespread Petunia axillaris and the highly endemic and endangered P. exserta. RESULTS Using population genetics, Bayesian linear mixed modelling and genome-wide association studies, we found that the three pollination syndrome traits vary in genetic architecture. Few genome regions explain a majority of the variation in flavonol content (defining UV floral colour) and strongly predict the trait value in hybrids irrespective of interspecific admixture in the rest of their genomes. In contrast, variation in pistil exsertion and anthocyanin content (defining visible floral colour) is controlled by many genome-wide loci. Opposite to flavonol content, the genome-wide proportion of admixture between the two species predicts trait values in their hybrids. Finally, the genome regions strongly associated with the traits do not show extreme divergence between individuals representing the two species, suggesting that divergent selection on these genome regions is relatively weak within their contact zones. CONCLUSIONS Among the traits analysed, those with a more complex genetic architecture are best maintained in association with the species upon their secondary contact. We propose that this maintained genotype-phenotype association is a coincidental consequence of the complex genetic architectures of these traits: some of their many underlying small-effect loci are likely to be coincidentally linked with the actual barrier loci keeping these species partially isolated upon secondary contact. Hence, the genetic architecture of a trait seems to matter for the outcome of hybridization not only then when the trait itself is under selection

Genetic diversity in Calibrachoa pygmaea (Solanaceae) a hawkmoth-pollinated nightshade from the Pampas

Calibrachoa pygmaea is a unique species of Calibrachoa, especially concerning its flower morphology and the environment where it occurs. The species is self-incompatible and is narrowly distributed in wet and flooded fields of the Pampas region. We characterize the genetic diversity of the species based on traditional plastid markers and newly developed nuclear microsatellites to identify drivers that guide its evolution. Our results identified markers that are informative and useful for studying the population structure of C. pygmaea, as well as that of other species of Calibrachoa. Both marker sets were congruent in developing conclusions regarding the evolutionary scenario of C. pygmaea, and revealed that the genetic variability and population structure of the species could be explained by common allele fixation or shared ancestral polymorphism, while its diversification can be attributed mainly to the species’ dispersal ability and certain ecological features

Genetic architecture of a pollinator shift and its fate in secondary hybrid zones of two Petunia species

Background: Theory suggests that the genetic architecture of traits under divergent natural selection influences how easily reproductive barriers evolve and are maintained between species. Divergently selected traits with a simple genetic architecture (few loci with major phenotypic effects) should facilitate the establishment and maintenance of reproductive isolation between species that are still connected by some gene flow. While empirical support for this idea appears to be mixed, most studies test the influence of trait architectures on reproductive isolation only indirectly. Petunia plant species are, in part, reproductively isolated by their different pollinators. To investigate the genetic causes and consequences of this ecological isolation, we deciphered the genetic architecture of three floral pollination syndrome traits in naturally occurring hybrids between the widespread Petunia axillaris and the highly endemic and endangered P. exserta. Results Using population genetics, Bayesian linear mixed modelling and genome-wide association studies, we found that the three pollination syndrome traits vary in genetic architecture. Few genome regions explain a majority of the variation in flavonol content (defining UV floral colour) and strongly predict the trait value in hybrids irrespective of interspecific admixture in the rest of their genomes. In contrast, variation in pistil exsertion and anthocyanin content (defining visible floral colour) is controlled by many genome-wide loci. Opposite to flavonol content, the genome-wide proportion of admixture between the two species predicts trait values in their hybrids. Finally, the genome regions strongly associated with the traits do not show extreme divergence between individuals representing the two species, suggesting that divergent selection on these genome regions is relatively weak within their contact zones. Conclusions: Among the traits analysed, those with a more complex genetic architecture are best maintained in association with the species upon their secondary contact. We propose that this maintained genotype–phenotype association is a coincidental consequence of the complex genetic architectures of these traits: some of their many underlying small-effect loci are likely to be coincidentally linked with the actual barrier loci keeping these species partially isolated upon secondary contact. Hence, the genetic architecture of a trait seems to matter for the outcome of hybridization not only then when the trait itself is under selection