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

Genetic diversity and population structure of naturally rare Calibrachoa species with small distribution in southern Brazil

Calibrachoa is a South-American genus comprising 27 species, several considered endemic or rare; few were subjects in genetic studies. We attempted to generate new data about the phylogenetically related and rare species C. eglandulata, C. sendtneriana, C. serrulata, and C. spathulata concerning their genetic diversity and population structure, which, coupled with their known restricted distribution, could help access their conservation status and contribute to the study of the Brazilian biodiversity. We sequenced 88 individuals for plastid intergenic spacers and genotyped 186 individuals for five microsatellite loci. Compared among each other, C. sendtneriana and C. serrulata presented the highest values of genetic diversity [% (sd) = 0.23 (0.14) and 0.43 (0.25), respectively], followed by C. spathulata [% (sd) = 0.19 (0.12)] and C. eglandulata [% (sd) = 0.02 (0.03)]. Population differentiation was evident for these latter species, whereas it was not significant for C. sendtneriana and C. serrulata. Factors such as habitat specificity and fragmentation, pollination syndrome, and life history could explain the observed patterns. Based on the new genetic data and the species’ biology, a conservation status was assigned for C. sendtneriana and the status of the other three species was reviewed

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

Landscape and climatic features drive genetic differentiation processes in a South American coastal plant

Background: Historical and ecological processes shape patterns of genetic diversity in plant species. Colonization to new environments and geographical landscape features determine, amongst other factors, genetic diversity withinand diferentiation between-populations. We analyse the genetic diversity and population structure of Calibrachoa heterophylla to infer the infuence of abiotic landscape features on the level of gene fow in this coastal species of the South Atlantic Coastal Plain. Results: The C. heterophylla populations located on early-deposited coastal plain regions show higher genetic diversity than those closer to the sea. The genetic diferentiation follows a pattern of isolation-by-distance. Landscape features, such as water bodies and wind corridors, and geographical distances equally explain the observed genetic diferentiation, whereas the precipitation seasonality exhibits a strong signal for isolation-by-environment in marginal populations. The estimated levels of gene fow suggest that marginal populations had restricted immigration rates enhancing diferentiation. Conclusions: Topographical features related to coastal plain deposition history infuence population diferentiation in C. heterophylla. Gene fow is mainly restricted to nearby populations and facilitated by wind felds, albeit without any apparent infuence of large water bodies. Furthermore, diferential rainfall regimes in marginal populations seem to promote genetic diferentiation

Characterization of mandarin citrus germplasm from Southern Brazil by morphological and molecular analyses

O objetivo deste trabalho foi caracterizar o germoplasma de tangerineiras (Citrus spp.) por meio de análises morfológicas e moleculares. Trinta e sete cultivares, pertencentes a 34 variedades distintas de tangerineiras, foram avaliadas pelas características morfológicas e agronômicas de folhas, flores e frutos, bem como por análises de microssatélites. As características morfológicas e agronômicas sugerem que praticamente todas as variedades podem ser usadas comercialmente e algumas delas, como a variedade Satsuma, são recomendadas para uso em cruzamentos dirigidos. O DNA foi obtido de amostras extraídas de uma a cinco plantas de cada cultivar. Oito de nove primers detectaram polimorfismos. Foram encontrados marcadores específicos para alguns acessos. O dendrograma construído com os resultados morfológicos dividiu as 37 cultivares em quatro grupos, enquanto o obtido com os microssatélites agrupou 35 das 37 cultivares em três grupos. As diferenças intervarietais não são grandes, e esta falta de concordância nas duas análises multivariadas indica que fatores evolutivos diversos estão agindo nesses dois níveis de investigação.The objective of this work was to characterize mandarin (Citrus spp.) germplasm from Southern Brazil by morphological and molecular analyses. Thirty seven cultivars from 34 distinct mandarin varieties were evaluated by morphological and agronomic traits of leaves, flowers and fruits, and by microsatellite markers. The morphological and agronomic characteristics suggested that almost all varieties can be produced for commercial use, and some, as the Satsuma variety, are recommended for breeding programs. Pooled DNA samples from 1-5 plants belonging to each cultivar were tested. Eight of the nine primers detected polymorphisms. Specific markers were found for some accessions. The dendrogram constructed with the morphological results divided the 37 cultivars into four groups, while that obtained with the microsatellites clustered 35 of the 37 cultivars into three groups only. Generally, intervarietal differences are not high, and this lack of agreement in the two multifactorial analyses indicates that diverse evolutionary factors are acting at these two levels of investigation

Could the reproductive system explain the stability and long-term persistence in a natural hybrid zone of Petunia (Solanaceae)?

The long-term success of populations is dependent on individual reproductive success, and in general, increased population size and genetic diversity contribute to population maintenance, reducing the risk of local extinction. Interspecific hybridization has consequences that can vary according to the hybrids' fate, which can be strongly influenced by the reproductive capacity of hybrids and canonical individuals from the contact zone. We examined the reproductive biology and morphology of two closely related Petunia (Solanaceae) species and their interspecific hybrids from Serra do Sudeste, Southern Brazil, and we measured their reproductive success under controlled conditions based on seed production and germination from five pollination treatments. We found differences in self-compatibility degree among individuals, lineages, and pollination treatments based on high total seed production (> 204,000 seeds) and germination (630 seedlings evaluated). No correlation was observed between corolla colour and reproductive success or between floral morphological traits and compatibility. High self-compatibility and inter-lineage compatibility can explain the hybrid populations' maintenance and origin, favouring the two analysed species' introgression