Fabio M. Sene

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.

Systematics and Evolution of the Drosophila buzzatii (Diptera: Drosophilidae) Cluster Using mtDNA

Abstract The Drosophila buzzatii Patterson & Wheeler cluster (repleta group) includes seven species: D. buzzatii Patterson & Wheeler, D. koepferae Fontdevila et al. D. serido Vilela & Sene, D. seriema Tidon-Sklorz & Sene, D. borborema Vilela & Sene, D. sp. D and D. sp. B. These flies are widely distributed in South America outside the Amazon region. The systematics of this cluster has been based on chromosomal inversions and the aedeagus is used to identify species. These species use necrotic cactus tissues as breeding sites. The current hypothesis of differentiation and speciation of these species is related to expansion and retraction of cactus distribution in South America during Quaternary climatic cycles. We investigated the phylogenetic relationship among species of this cluster based on the mtDNA COI gene region and compared it with the relationship established using classical markers. The resulting phylogenetic hypothesis indicated that this cluster is a monophyletic group that can be divided into two sets of species: the one including D. buzzatii and D. koepferae and other with the remaining five species. The latter can also be divided into two clades. Although this branching pattern is similar to the one established by classical markers, some disagreement involving populations was observed that suggests secondary contact between populations of different species. The distribution pattern of COI haplotypes is partitioned geographically, which could be the result of limited gene flow between groups of species suggesting a longer history of differentiation than previously hypothesized for D. buzzatii cluster species.

Heterochromatin and karyotypic differentiation of some neotropical cactus-breeding species of the Drosophila repleta species group

Metaphase chromosomes from neuroblasts of strains from both laboratory stocks and natural populations of D. serido, D. meridionalis, D. borborema and D. buzzatii have been studied using colcemid pretreatment, and air-drying followed by Giemsa staining. The two latter species both show a uniform ‘basic’ metaphase karyotype. D. serido and D. meridionalis, on the other hand, both include a number of different, geographically distinct, metaphase karyotypes involving differences in the major blocks of constitutive heterochromatin present on the sex chromosomes and/or the 6th chromosome (microchromosome). These chromosomal differences are largely due to the acquisition of extra heterochromatin though pericentric inversions appear to be responsible for some of the Y-chromosome variants in D. serido. Moreover, the cytological evidence demonstrates that populations of both species are far from continuous in distribution. The extent to which such cytological differences reflect the existence of subspecific or specific complexes with minimal morphological differentiation is under investigation.

Two new species of the Drosophila serido sibling set (Diptera, Drosophilidae)

Drosophila antonietae sp. nov. and D. gouveai sp. nov. are members of the D. buzzatii cluster of the D. repleta species group of the genus Drosophila. They can be distinguished from their cryptic species, D. borborema Vilela & Sene, 1977, D. koepferae Fontdevila & Wasserman, 1988, D. serido Vilela & Sene, 1977, and D. seriema Tidon-Sklorz & Sene, 1995 by morphological, genetic and ecological criteria.

Sequence analysis, chromosomal distribution and long-range organization show that rapid turnover of new and old pBuM satellite DNA repeats leads to different patterns of variation in seven species of the Drosophila buzzatii cluster

We aimed to study patterns of variation and factors influencing the evolutionary dynamics of a satellite DNA, pBuM, in all seven Drosophila species from the buzzatii cluster (repleta group). We analyzed 117 alpha pBuM-1 (monomer length 190 bp) and 119 composite alpha/beta (370 bp) pBuM-2 repeats and determined the chromosome location and long-range organization on DNA fibers of major sequence variants. Such combined methodologies in the study of satDNAs have been used in very few organisms. In most species, concerted evolution is linked to high copy number of pBuM repeats. Species presenting low-abundance and scattered distributed pBuM repeats did not undergo concerted evolution and maintained part of the ancestral inter-repeat variability. The alpha and alpha/beta repeats colocalized in heterochromatic regions and were distributed on multiple chromosomes, with notable differences between species. High-resolution FISH revealed array sizes of a few kilobases to over 0.7 Mb and mutual arrangements of alpha and alpha/beta repeats along the same DNA fibers, but with considerable changes in the amount of each variant across species. From sequence, chromosomal and phylogenetic data, we could infer that homogenization and amplification events involved both new and ancestral pBuM variants. Altogether, the data on the structure and organization of the pBuM satDNA give insights into genome evolution including mechanisms that contribute to concerted evolution and diversification.

The Drosophila serido speciation puzzle: putting new pieces together.

The D. seridosuperspecies is a complex mosaic of populations distributed over a vast part of South America and showing various degrees of genetical divergence. We have analyzed its chromosomal constitution in 16 new localities of southeastern and southern Brazil. Both the metaphase and salivary gland chromosomes show a sharp split of these populations in two groups. Four populations, fixed for inversion 2e8and showing the type I karyotype, represent the southwestern limit of D. seridotype B, which inhabits the Cerrado in central-western Brazil. The remaining populations are homozygous for 2x7, an inversion also fixed in the Caatinga populations of northeastern Brazil. However, their karyotype, in those populations analyzed, belong to a different type (V) from that of the Caatinga populations. Populations in this second group are polymorphic for five inversions on chromosome 2 plus another on chromosome 5 and show considerable interpopulation differentiation. The breakpoints of chromosome 2 inversions are described and the inversion loops of several heterokaryotypes are presented. Biogeographical information suggests that there are clear ecological differences between the two groups of populations as well as among the populations within the second group. The possible role of host plants in promoting the genetic divergence among the D. seridopopulations is discussed.

Population Genetic Structure of Two Columnar Cacti with a Patchy Distribution in Eastern Brazil

The genetic variability and population genetic structure of six populations of Praecereus euchlorus and Pilosocereus machrisii were investigated. The genetic variability in single populations of Pilosocereus vilaboensis, Pilosocereus aureispinus, and Facheiroa squamosa was also examined. All of these cacti species have a patchy geographic distribution in which they are restricted to small areas of xeric habitats in eastern Brazil. An analysis of genetic structure was used to gain insights into the historical mechanisms responsible for the patchy distribution of P. euchlorus and P. machrisii. High genetic variability was found at the populational level in all species (P=58.9–92.8%, Ap=2.34–3.33, He=0.266–0.401), and did not support our expectations of low variability based on the small population size. Substantial inbreeding was detected within populations (FIS=0.370–0.623). In agreement with their insular distribution patterns, P. euchlorus and P. machrisii had a high genetic differentiation (FST=0.484 and FST=0.281, respectively), with no evidence of isolation by distance. Accordingly, estimates of gene flow (Nm) calculated from FST and private alleles were below the level of Nm=1 in P. machrisii and P. euchlorus. These results favored historical fragmentation as the mechanism responsible for the patchy distribution of these two species. The genetic distance between P. machrisii and P. vilaboensis was not compatible with their taxonomic distinction, indicating a possible local speciation event in this genus, or the occurrence of introgression events.

Preliminary data on the geographical distribution of Drosophila species within morphoclimatic domains of Brazil. III. The cardini group

A reanalysis, based on museum specimens, of our previously published data on the geographical distribution of the species of Drosophila belonging to the cardini group in Brazil is presented and discussed. As previously recorded in several papers, including ours, the following four species were recognized: D. cardini, D. cardinoides, D. neocardini, and D. polymorpha. However, it was realized that most of the flies we have previously identified as Drosophila cardinoides belong in fact to Drosophila cardini. To facilitate the proper identification of these four near-sibling species, their holotypes were analyzed and their terminalia were described and illustrated. A key to the four species is also provided.

Tooth structure and fracture strength of cavities

This study evaluated, in vitro, the loss of tooth substance after cavity preparation for direct and indirect restorations and its relationship with fracture strength of the prepared teeth. Sixty sound human maxillary first premolars were assigned to 6 groups (n=10). MOD direct composite cavities (Groups I, II and III) and indirect inlay cavities (Groups IV, V and VI) were prepared maintaining standardized dimensions: 2-mm deep pulpal floors, 1.5-mm wide gingival walls and 2-mm high axial walls. Buccolingual width of the occlusal box was established at 1/4 (Groups I and IV), 1/3 (Groups II and V) or 1/2 (Groups III and VI) of the intercuspal distance. Teeth were weighed (digital balance accurate to 0.001 g) before and after preparation to record tooth substance mass lost during cavity preparation. The prepared teeth were submitted to occlusal loading to determine their fracture strength using a universal testing machine at a crosshead speed of 0.5 mm/min. Data were analyzed by two-way ANOVA and Tukey test (alpha= 0.05). 1/4-inlay cavities had higher percent mean mass loss (9.71%) than composite resin cavities with the same width (7.07%). 1/3-inlay preparations also produced higher percent mean mass loss (13.91%) than composite resin preparations with the same width (10.02%). 1/2-inlay cavities had 21.34% of mass loss versus 16.19% for the 1/2-composite resin cavities. Fracture strength means (in kgf) were: GI = 187.65; GII = 143.62; GIII = 74.10; GIV = 164.22; GV = 101.92; GVI = 50.35. Statistically significant difference (p<0.05) were observed between Groups I and IV, II and V, III and VI. Higher tooth structure loss and lower fracture strength were recorded after preparation of inlay cavities, regardless of the width of the occlusal box, compared to the direct composite resin cavities.

Characterisation and interpopulation variability of a complex HpaI satellite DNA of Drosophila seriema (repleta group).

A HpaI satellite DNA has been isolated and characterised from the genome of Drosophila seriema, a cactus-breeding species endemic to the rock fields of the Espinhaço Range in Brazil. The monomer sequences are slightly A + T rich (66%) and there is a significant variation of repetition length (343–391 bp). The length variability is mainly due to a 22 bp indel in some repeats and the presence of a highly variable region characterised by several DNA rearrangements, including indels, inversions and duplications of small sequence segments. The retarded mobility of monomers observed after gel electrophoresis suggests DNA curvature. Thirty satDNA repeats were analysed in samples from five populations which cover D. seriema geographical distribution. Previous studies showed that these populations present low levels of chromosomal divergence in contrast to high levels of mtDNA divergence. The variability among the 30 repeats is pretty low, on average 2%. The results showed that the satDNA sequences are rather homogeneous on both intra and interpopulational levels, presenting no specific feature(s) that could discriminate a particular population or groups of geographically close populations. Possible factors responsible for such homogeneity are discussed.