Phylogeography of Drosophila buzzatii (Diptera, Drosophilidae): responses of the species to Quaternary climates in tropical and subtropical South America.

Drosophila buzzatii (Diptera: Drosophilidae) is a fly that breeds exclusively on decaying tissues of cacti species widely distributed in tropical and subtropical areas of South America. This distribution includes biomes in distinct climatic regimes (e.g., seasonal rain forest, semi-arid scrubs, savannas, and grasslands), which at first glance could might give the false impression that the species is not sensitive to either climate or vegetation physiognomies. However, detection of historical demographic events within D. buzzatii reveal the interplay between climate and the population structure of the species as the Late Quaternary climate changes occurred. To understand this process, we performed a phylogeographic analysis based on sequences of the mitochondrial gene COI for 128 individuals from 43 localities. Our analyses combined coalescent methods, population genetics, and paleodistributions estimation methods. Our study reveals that the COI haplotype diversity is geographically structured, with a decreasing cline from north to south. The results suggest an ancient range expansion, dated from 610k to 550k years before present, in the northernmost region of the species distribution, the Caatinga vegetation. More recently, an intense gene flow and a population expansion were detected in the central and south portions of its distribution. The demographic events detected date back to the glacial periods of the Quaternary.

A holocenic and dynamic hybrid zone between two cactophilic Drosophila species in a coastal lowland plain of the Brazilian Atlantic Forest.

Hybridization and introgression are processes that contribute to shaping biological diversity. The factors promoting the formation of these processes are multiples but poorly explored in a biogeographical and ecological context. In the southeast coastal plain of the Brazilian Atlantic Forest, a hybrid zone was described between two closely related cactophilic species, Drosophila antonietae and D. serido. Here, we revisited and analysed specimens from this hybrid zone to evaluate its temporal and spatial dynamic. We examined allopatric and sympatric populations of the flies using independent sources of data such as mitochondrial and nuclear sequences, microsatellite loci, morphometrics of wings and male genitalia, and climatic niche models. We also verified the emergence of the flies from necrotic tissues of collected cacti to verify the role of host association for the population dynamics. Our results support the existence of a hybrid zone due to secondary contact and limited to the localities where the two species are currently in contact. Furthermore, we detected asymmetric bidirectional introgression and the maintenance of the species integrity, ecological association and morphological characters, suggesting selection and limited introgression. Considering our paleomodels, probably this hybrid zone is recent and the contact occurred during the Holocene to the present day, favoured by range expansion of their populations due to expansion of open and dry areas in eastern South America during palaeoclimatic and geomorphological events.

Plastic Variation in the Phyletic Lineages of Cactophilic Drosophila meridionalis and Relation to Hosts as Potential for Diversification.

The insect/plant interaction is known to be a trigger for diversification and even speciation. Experimental analyses on fitness traits and phenotypic variation using alternative host sites have been performed to understand the process of diversification relative to insect/plant interactions. For cactophilic species of Drosophila, the speciation process is considered an adaptive radiation in response to the exploration of species of the Cactaceae as breeding and feeding sites. In this work, we analyzed life history and morphological traits in individuals from two phyletic lineages (Evolutionarily Significant Units ESU) of the cactophilic species Drosophila meridionalis (Wasserman 1962) (Diptera: Drosophilidae) raised from media prepare. The characters analyzed corresponded to viability, developmental time, and four morphological measurements. The experiments were performed in a semi-natural medium prepared with fermenting tissues of the natural hosts, Cereus hildmaniannus and Opuntia monacantha. Viability, development time, and three morphological measurements were influenced by lineage, suggesting differentiation between the lineages. However, in O. monacantha, the mean viability was greater (~15%) and development time was longer (~336 h) than in C. hildmaniannus (~11% and ~301 h, respectively). Only the developmental time was significantly affected by the host cactus. In general, ESU group A had better values than ESU group BC for the evaluated traits. This finding suggested differentiation between the two lineages and different plastic responsiveness to the contrasting environments of the hosts, and that C. hildmaniannus may be a relatively stressful environment for the larvae, as for other Drosophila species.

Genetics and Distribution Modeling: The Demographic History of the Cactophilic Drosophila buzzatii Species Cluster in Open Areas of South America.

Biodiversity is the result of historical and recurrent events acting on populations and species. The Drosophila buzzatii species cluster is distributed along a diagonal of open areas in South America. Combining genetic analyses with species distribution models we evaluated the influence of climatic changes in the demography history of this cluster. We performed a Bayesian Skyline analysis and reconstructed the ancestral areas based on mitochondrial cytochrome oxidase I (mtCOI) gene. We modeled the current distribution and projected it to past (mid-Holocene and Last Glacial Maximum) and future. Our results demonstrate that climate change plays a critical role in historical demography and in defining the current and future geographic ranges of these species. The inter-Andean dry valleys and the Chiquitano Seasonally Dry Tropical Forests (SDTF) in Bolivia are considered the ancestral area for the D. buzzatii cluster. From this area, the migration route was through a west-east corridor from central Andes, throughout Bolivia, Paraguay and Argentina toward eastern and northeastern Brazil, along the edges of rain forest. The responses of the species to the climatic changes differ from the dominant assumptions of expansion during dry/cold weather and contraction during wet/warm weather that characterized the glacial cycles. We suggest that the influence of ecology and the potential responsiveness of each taxon to the environmental dynamics should be considered as well. Predictions for the future suggest a decline of suitable areas for the cluster, threatening biodiversity of these habitats. This work showed the importance of an integrative analysis of genetics and geography information to improve the inferences about demographic history hypotheses for the cluster.

Low satellite DNA variability in natural populations of Drosophila antonietae involved in different evolutionary events.

Drosophila antonietae is a cactophilic species that is found in the mesophilic forest of the Paraná-Paraguay river basin and in the dunes of the South Atlantic coast of Brazil. Although the genetic structure of the Paraná-Paraguay river basin populations has already been established, the relationship between these populations and those on the Atlantic coast is controversial. In this study, we compared 33 repetitive units of pBuM-2 satellite DNA isolated from individuals from 8 populations of D. antonietae in these geographic regions, including some populations found within a contact zone with the closely related D. serido. The pBuM-2 sequences showed low interpopulational variability. This result was interpreted as a consequence of both gene flow among the populations and unequal crossing over promoting homogenization of the tandem arrays. The results presented here, together with those of previous studies, highlight the use of pBuM-2 for solving taxonomic conflicts within the D. buzzatii species cluster.

Curvature measurement as a tool for the analysis of morphometric variation using Drosophila wings as a model

This article describes the use of geometric measurements of continuous, discrete parameters to studymorphometric variation in the wing cells of two sibling species, Drosophila mercatorum and Drosophilaparanaensis. To validate the results, the same wing samples were also analyzed using truss networks andpartial warps, in addition to a comparison with the ellipse method. The use of discriminative measurementsin conjunction with a Bayesian-based classification method yielded a relatively high number of correctclassifications for new individuals. These results compared favorably with those obtained using trussnetworks, partial warps and the ellipse method. These findings indicate that continuous curvature and arclength measurements may be useful parameters for the morphometric analysis of insect wings and possiblyother biological structures and shapes.

Conservation of pBuM-2 satellite DNA sequences among geographically isolated Drosophila gouveai populations from Brazil.

In this study, we have compared 34 repetition units of pBuM-2 satellite DNA of individuals from six isolated populations of Drosophila gouveai, a cactophilic member of Drosophila buzzatii cluster (repleta group). In contrast to the results of previous morphological and molecular data, which suggest differentiation among the D. gouveai populations, the sequences and the cluster analysis of pBuM-2 monomers showed that this repetitive element is highly conserved among the six D. gouveai populations (97.8% similarity), indicating a slow rate of evolution of pBuM-2 sequences at the population level. Probably, some homogenization mechanisms of tandem sequences, such as unequal crossing or gene conversion, have maintained the sequence similarity of pBuM-2 among D. gouveai populations. Alternatively, such a result may be associated with a functional role of pBuM-2 sequences, although it is not understood at present.

Cactophilic Drosophila in South America: a model for evolutionary studies.

The Drosophila buzzatii cluster is composed of seven cactophilic species and their known geographical distribution encompasses the open vegetation diagonal, which includes the morphoclimatic Domains of the Caatinga, Chaco and Cerrado, which are situated between the Amazon and the Atlantic forests. Besides these areas, these cactophilic species are also found in a narrow strip along the Atlantic coast from northeastern Brazil to the southern tip of the country. The hypothesis of vicariant events, defining the core areas of each species, is proposed to explain the historical diversification for the cluster. The intraspecific analysis for the cluster shows a population structure with gene flow restricted by distance, range expansion with secondary contact resulting in introgression and simpatry, especially in the limits of the species distribution, polytypic populations and assortative mating in inter population experiments. There is a variation related to these events that depends on the species and geographic origin of the population analyzed. These events are, hypothetically, described as the results of expansion and retraction of the population ranges, as a consequence of their association with cacti, which theoretically follow the expansion and retraction of dry areas during the paleoclimatic oscillations in South America, as that promoted by the glacial cycles of the Quaternary. The Drosophila buzzatii cluster is divided into two groups. The first one is composed of D. buzzatii, a species that has a broad geographic distribution and no significant differentiation between its populations. The second is the Drosophila serido sibling set, which encompasses the others species and is characterized by a significant potential for differentiation.

Molecular characterization and chromosomal distribution of Galileo, Kepler and Newton, three foldback transposable elements of the Drosophila buzzatii species complex.

Galileo is a foldback transposable element that has been implicated in the generation of two polymorphic chromosomal inversions in Drosophila buzzatii. Analysis of the inversion breakpoints led to the discovery of two additional elements, called Kepler and Newton, sharing sequence and structural similarities with Galileo. Here, we describe in detail the molecular structure of these three elements, on the basis of the 13 copies found at the inversion breakpoints plus 10 additional copies isolated during this work. Similarly to the foldback elements described in other organisms, these elements have long inverted terminal repeats, which in the case of Galileo possess a complex structure and display a high degree of internal variability between copies. A phylogenetic tree built with their shared sequences shows that the three elements are closely related and diverged approximately 10 million years ago. We have also analyzed the abundance and chromosomal distribution of these elements in D. buzzatii and other species of the repleta group by Southern analysis and in situ hybridization. Overall, the results suggest that these foldback elements are present in all the buzzatti complex species and may have played an important role in shaping their genomes. In addition, we show that recombination rate is the main factor determining the chromosomal distribution of these elements.