Saltans Group Research Articles

Phylogeny of Drosophila saltans group (Diptera: Drosophilidae) based on morphological and molecular evidence.

Drosophila saltans group belongs to the subgenus Sophophora (family Drosophilidae), and it is subdivided into five subgroups, with 23 species. The species in this group are widely distributed in the Americas, primarily in the Neotropics. In the literature, the phylogenetic reconstruction of this group has been performed with various markers, but many inconsistencies remain. Here, we present a phylogenetic reconstruction of the saltans group with a greater number of species, 16 species, which is the most complete to date for the saltans group and includes all subgroups, in a combined analysis with morphological and molecular markers. We incorporated 48 morphological characters of male terminalia, the highest number used to date, and molecular markers based on mitochondrial genes COI and COII. Based on the results, which have recovered the five subgroups as distinct lineages, we propose a new hypothesis regarding the phylogenetic relationships among the subgroups of the saltans group. The relationships of the species within the sturtevanti and elliptica subgroups were well supported. The saltans subgroup showed several polytomies, but the relationship between the sibling species D. austrosaltans and D. saltans and their close relation with D. nigrosaltans were well supported in the molecular and total evidence analyses. The morphological analysis additionally supported the formation of the clade D. nigrosaltans—D. pseudosaltans. The observed polytomies may represent synchronous radiations or have resulted from speciation rates that have been too fast relative to the pace of substitution accumulation.

Male terminalia morphology of sixteen species of the Drosophila saltans group Sturtevant (Diptera, Drosophilidae).

Male terminalia in insects with internal fertilization evolve more rapidly than other structures. The aedeagus is the most variable structure, making it a valuable diagnostic feature to distinguish species. The saltans group Sturtevant of Drosophila Fall contains sibling species, that can be distinguished by their aedeagi. Here, we revised and illustrated the morphology of the male terminalia of the following species: Drosophila prosaltans Duda, 1927; D. saltans Sturtevant, 1916; D. lusaltans Magalhes, 1962; D. austrosaltans Spassky, 1957; D. septentriosaltans Magalhes, 1962; D. nigrosaltans Magalhes, 1962; D. pseudosaltans Magalhes, 1956; D. sturtevanti Duda, 1927; D. lehrmanae Madi-Ravazzi et al., 2021; D. dacunhai Mouro Bicudo, 1967; D. milleri Magalhes, 1962; D. parasaltans Magalhes, 1956; D. emarginata Sturtevant, 1942; D. neoelliptica Pavan Magalhes in Pavan, 1950; D. neosaltans Pavan Magalhes in Pavan, 1950 and D. neocordata Magalhes, 1956. We found that phallic structures (e.g., the aedeagus) evolve more rapidly than periphallic structures (e.g., epandrium), being completely different among the subgroups and within them. This rapid evolution may be due to the action of sexual selection or to the potential role of those structures in speciation.

Integrative taxonomy and a new species description in the sturtevanti subgroup of the Drosophila saltans group (Diptera: Drosophilidae)

Although the biological concept of species is well established in animals, sometimes the decision about the specific status of a new species is difficult and hence requires support of an integrative analysis of several character sets. To date, the species Drosophila sturtevanti, D. magalhaesi, D. milleri and D. dacunhai, belonging to the sturtevanti subgroup of the Neotropical saltans species group, are identified mainly by the aedeagus morphology, but also present some differences in spot coloration and patterning of the female sixth tergite and in the shape and size of the spermathecae, parallel to a pattern of reproductive isolation. In the present study, we describe a novel saltans group species from French Guiana belonging to the sturtevanti subgroup. Our species designation is based on an integrative approach covering (i) aedeagi and spermathecae morphology by scanning electron microscopy, (ii) analysis of female sixth-tergite color, (iii) morphometrical analysis of aedeagi and wings, (iv) analysis of partial sequence of the COI, COII and ND4 mitochondrial genes as well as (v) intercrosses for analysis of reproductive isolation. The comparative analysis of the results on these markers with those of D. sturtevanti, D. milleri and D. dacunhai supports that this line belongs to a new species of the sturtevanti subgroup that we name Drosophila lehrmanae sp. nov. in honor of Prof. Lee Ehrman´s 85th birthday.

Research Article Development and transferability of microsatellite markers for species of the <i>saltans</i> group of <i>Drosophila</i> (Diptera: Drosophilidae)

The microsatellite markers (SSRs) are highly polymorphic, fast evolving, and regarded as neutral markers. Due to these traits, they have been widely used in population studies. The development of specific SSRs for a given species opens possibilities for the application, by means of transferability, in population studies of other species that are phylogenetically related, thereby optimizing time and cost. This study describes, for the first time, 16 primers pairs developed for Drosophila sturtevanti, which were tested their transferability for 14 species of the saltans group of Drosophila. The adequate conditions of amplification were established by using 15 males of D. sturtevanti. The total percentage of transfer has been 49.45%. The species with the highest success rates of heterologous amplification have been included in the sturtevanti subgroup. Two microsatellite markers have amplified in all the species, whilst one would not amplify in any of the species of the saltans group. The data corroborate the use of the technique for the transferability of these oligonucleotides, which may be used in studies of genetic diversity and population structure of D. sturtevanti and other species of the saltans group of Drosophila

Scanning electron microscopy of male terminalia and its application to species recognition and phylogenetic reconstruction in the Drosophila saltans group.

The Drosophila saltans group consists of five subgroups and 21 species, most of which have been identified only by morphological aspects of the male terminalia revealed by drawings using a camera lucida and a bright-field microscope. However, several species in the group, mainly those included in the saltans subgroup, are difficult to differentiate using only these characteristics. In this study, we used scanning electron microscopy (SEM) to analyze 19 structures of the male terminalia in 10 species from the five saltans subgroups. Among these structures, nine could be identified only through SEM analysis. We aimed to find other characteristics useful for morphological recognition of these species and to use these characteristics for phylogenetic reconstruction. These morphological differences enabled us to effectively distinguish among sibling species. These findings confirmed the monophyly of this group as previously determined in evolutionary studies based on other markers. The single most parsimonious tree (CI = 87 and RI = 90) indicated that the cordata subgroup is the most basal lineage and the saltans subgroup is the most apical lineage, as shown in earlier studies based on morphological data. However, our findings differed somewhat from these studies with respect to the phylogenetic relationships of species in the saltans group indicating that this group is still a puzzle that remains to be deciphered.

Harrow: new Drosophila hAT transposons involved in horizontal transfer

In this study we characterize the transposable elements harrow, which belong to the hAT superfamily of DNA transposons. Searches for harrow sequences were performed in 65 Drosophilidae species, mainly representing Neotropical and cosmopolitan groups from the genus Drosophila. The nucleotide divergence among elements found in these species suggests that harrow sequences could be clustered in a subfamily. The patchy distribution throughout the genus Drosophila and the high similarity presented between all harrow sequences indicate that horizontal transfer could play a major role in the evolution of harrow elements. The results obtained suggest an evolutionary scenario in which harrow would have undergone multiple horizontal transfer events in the Neotropics, involving D. tripuncatata, D. mojavensis (Subgenus Drosophila) and several species of the willistoni and saltans groups (subgenus Sophophora).

Phylogenetic Relationships Among Species Subgroups in the <I>Drosophila saltans</I> Group (Diptera: Drosophilidae):Can Morphology Solve a Molecular Conflict?

Proper phylogenetic reconstruction is crucial for understanding many evolutionary phenomena. In spite of the great success of molecular phylogenetics, DNA signal still may be limited by some intrinsic constraints such as codon usage bias. The phylogenetic relationships between the five species subgroups of the Drosophila saltans group are a good example of conflicting molecular phylogenies drawn from different genes due to an ancestral substitutional shift. Here, forty morphological characters were analyzed using the same set of species used in previous molecular studies, with at least a single representative of each subgroup. The cladistic analysis was in disagreement with most of the previous hypotheses in placing the sturtevanti subgroup as an early branch, whereas the four remaining subgroups form a well supported clade that can be further subdivided into two sister clades: one containing the cordata and the elliptica subgroups, whereas the second includes the parasaltans and the saltans subgroups. The molecular evolution (codon usage bias) of the saltans group were revised in light of the present finding. The analysis highlights the important role of morphology in phylogeny reconstruction and in understanding molecular evolutionary phenomena.

Variability of esterase patterns in adult flies of the saltans species group of Drosophila (subgenus Sophophora)

Esterases are known for their involvement in several physiological processes and high degree of polymorphism, in many organisms. Such polymorphism has been used to characterize species and species groups and to study genetic changes occurred in their evolutionary history. In the present study, the esterase patterns of 19 strains from 10 species representative of the five subgroups of the saltans species group were analyzed using polyacrylamide gel electrophoresis and alpha- and beta- naphthyl acetates as substrates. Fifty-one esterase bands were detected and classified as 31 alpha-esterases, 18 beta-esterases and two alpha/beta-esterases. On the basis of the inhibition patterns using Malathion and eserine sulfate, 34 bands were classified as carboxylesterases, 14 as acethylesterases and three as cholinesterases. Ten gene loci were tentatively established on the basis of data on band position in the gel, substrate preference and inhibition pattern. Twenty bands were species-specific, the remaining being shared by species from the same or different subgroups. Bands detected exclusively in males and bands with a different frequency or degree of expression between sexes were also detected. In the gels prepared for analysis of gene expression in the body parts (head, thorax and abdomen), the degree of expression of the beta-esterases was higher in the thorax, while the alpha-esterases were expressed predominantly in the abdomen and thorax. A global view of the data available at present on the esterases of the species from the saltans group and their degree of polymorphism are presented, as well as the possibility of using some beta-esterases, because of their characteristics in the gels, as markers for species identification.

Is the Evolutionary History of the O-Type P Element in the saltans and willistoni Groups of Drosophila Similar to That of the Canonical P Element?

We studied the occurrence of O-type P elements in at least one species of each subgroup of the saltans group, in order to better understand the phylogenetic relationships among the elements within the saltans group and with those of species belonging to the willistoni group. We found that the O-type subfamily has a patchy distribution within the saltans group (it does not occur in D. neocordata and D. emarginata), low sequence divergence among species of the saltans group as well as in relation to species of the willistoni group, a lower rate of synonymous substitution for coding sequences compared to Adh, and phylogenetic incongruities. These findings suggest that the evolutionary history of the O-type subfamily within the saltans and willistoni groups follows the same model proposed for the canonical subfamily of P elements, i.e., events of horizontal transfer between species of the saltans and willistoni groups.

Transposon display supports transpositional activity of P elements in species of the saltans group of Drosophila

Mobilization of two P element subfamilies (canonical and O-type) from Drosophila sturtevanti and D. saltans was evaluated for copy number and transposition activity using the transposon display (TD) technique. Pairwise distances between strains regarding the insertion polymorphism profile were estimated. Amplification of the P element based on copy number estimates was highly variable among the strains (D. sturtevanti, canonical 20.11, O-type 9.00; D. saltans, canonical 16.4, O-type 12.60 insertions, on average). The larger values obtained by TD compared to our previous data by Southern blotting support the higher sensitivity of TD over Southern analysis for estimating transposable element copy numbers. The higher numbers of the canonical P element and the greater divergence in its distribution within the genome of D. sturtevanti (24.8%) compared to the O-type (16.7%), as well as the greater divergence in the distribution of the canonical P element, between the D. sturtevanti (24.8%) and the D. saltans (18.3%) strains, suggest that the canonical element occupies more sites within the D. sturtevanti genome, most probably due to recent transposition activity. These data corroborate the hypothesis that the O-type is the oldest subfamily of P elements in the saltans group and suggest that the canonical P element is or has been transpositionally active until more recently in D. sturtevanti.

A New Nucleolar Body Appears inDrosophila saltansSalivary Gland Cells Before Histolysis, in Programmed Cell Death

The salivary glands of Drosophila saltans (saltans group, saltans subgroup) analyzed in an advanced stage of programmed cell death showed the appearance of a single, round, nucleolar corpuscle inside the highly altered nucleus of every gland cell, at a time during which the integrity of the original nucleolus was already lost and the original nucleolar material apparently disappeared. In the same nuclei, which already had also lost the characteristic chromosome structure, some delicate chromosome threads were maintained. In many cells, the new nucleolar corpuscle and these chromosome threads are associated. These findings are novel. However, the hypothesis put forward concerning their meaning remains dependent on other studies.

Regulation of Body Pigmentation by the Abdominal-B Hox Protein and Its Gain and Loss in Drosophila Evolution

Hox genes have been implicated in the evolution of many animal body patterns, but the molecular events underlying trait modification have not been elucidated. Pigmentation of the posterior male abdomen is a recently acquired trait in the Drosophila melanogaster lineage. Here, we show that the Abdominal-B (ABD-B) Hox protein directly activates expression of the yellow pigmentation gene in posterior segments. ABD-B regulation of pigmentation evolved through the gain of ABD-B binding sites in a specific cis-regulatory element of the yellow gene of a common ancestor of sexually dimorphic species. Within the melanogaster species group, male-specific pigmentation has subsequently been lost by at least three different mechanisms, including the mutational inactivation of a key ABD-B binding site in one lineage. These results demonstrate how Hox regulation of traits and target genes is gained and lost at the species level and have general implications for the evolution of body form at higher taxonomic levels.

Further Study on the Esterase Patterns of Sibling Species in the Drosophila saltans Subgroup (saltans Group): Intraspecific and Interspecific Variations in the Development

Twenty of the 32 esterase bands previously detected in the adults of D. prosaltans, D. saltans and D. austrosaltans were found in larvae and pupae studied in this work. The results showed that, in addition to expressing the highest number of esterase bands, the adult stage of the three species exhibited the highest degree of expression (amount of synthesis) for most of the bands. Differences between larval and pupal stages were detected in the degree of expression (amount of synthesis) of the bands and in the frequency of samples expressing them. The frequencies of expression of the bands corresponding to genes in loci 1-3 were greater in pupae than in larvae while the frequencies of expression of the bands corresponding to genes in loci 4-9 were predominantly expressed in larvae or were equal in both developmental stages. Like the adults, larvae, pupae and empty pupal cases (which were also studied in this work) showed specific esterases. Taken together, the observations showed that, in the species studied, every developmental stage is characterized by specific bands and by specific frequency and degree of expression of the bands shared with other stages.

Distribution and insertion numbers of transposable elements in species of the Drosophila saltans group

Abstract Information about the distribution and insertion numbers of many transposable elements is restricted to few speciesof Drosophila , although these elements are widely distributed throughout the genus . The aim of this work was to de-scribe the distribution and insertion numbers of four retrotransposons ( copia , gypsy , micropia , I ) and four trans-posons ( hobo , mariner , Minos and Bari-1 )inthe saltans group of Drosophila. Our data shows that, except for mariner , all the other elements are widespread within the saltans group and show variable insertion numbers of up to24 copies. Key words: Drosophila , genomic insertion, saltans group, retrotransposons, transposons.Received: March 11, 2005; Accepted: September 20, 2005. Introduction Transposable elements are a major component of thegenomes of most species and are widespread throughoutthe genus Drosophila. Most of our knowledge on trans-posable elements in Drosophila comes from studies car-ried out in natural populations of

Evolutionary dynamics of wAu-like Wolbachia variants in neotropical Drosophila spp.

Wolbachia bacteria are common intracellular symbionts of arthropods and have been extensively studied in Drosophila. Most research focuses on two Old Word hosts, Drosophila melanogaster and Drosophila simulans, and does not take into account that some of the Wolbachia associations in these species may have evolved only after their fast global expansion and after the exposure to Wolbachia of previously isolated habitats. Here we looked at Wolbachia of Neotropical Drosophila species. Seventy-one lines of 16 Neotropical Drosophila species sampled in different regions and at different time points were analyzed. Wolbachia is absent in lines of Drosophila willistoni collected before the 1970s, but more recent samples are infected with a strain designated wWil. Wolbachia is absent in all other species of the willistoni group. Polymorphic wWil-related strains were detected in some saltans group species, with D. septentriosaltans being coinfected with at least four variants. Based on wsp and ftsZ sequence data, wWil of D. willistoni is identical to wAu, a strain isolated from D. simulans, but can be discriminated when using a polymorphic minisatellite marker. In contrast to wAu, which infects both germ line and somatic tissues of D. simulans, wWil is found exclusively in the primordial germ line cells of D. willistoni embryos. We report on a pool of closely related Wolbachia strains in Neotropical Drosophila species as a potential source for the wAu strain in D. simulans. Possible evolutionary scenarios reconstructing the infection history of wAu-like Wolbachia in Neotropical Drosophila species and the Old World species D. simulans are discussed.

Multiple events of horizontal transfer of the Minos transposable element between Drosophila species

In this study the Minos element was analyzed in 26 species of the repleta group and seven species of the saltans group of the genus Drosophila. The PCR and Southern blot analysis showed a wide occurrence of the Minos transposable element among species of the repleta and the saltans groups and also a low number of insertions in both genomes. Three different analyses, nucleotide divergence, historical associations, and comparisons between substitution rates ( d N and d S) of Minos and Adh host gene sequences, suggest the occurrence of horizontal transfer between repleta and saltans species. These data reinforce and extend the Arcà and Savakis [Genetica 108 (2000) 263] results and suggest five events of horizontal transfer to explain the present Minos distribution: between D. saltans and the ancestor of the mulleri and the mojavensis clusters; between D. hydei and the ancestor of the mulleri and the mojavensis clusters; between D. mojavensis and D. aldrichi; between D. buzzatii and D. serido; and between D. spenceri and D. emarginata. An alternative explanation would be that repeated events of horizontal transfer involving D. hydei, which is a cosmopolitan species that diverged from the others repleta species as long as 14 Mya, could have spread Minos within the repleta group and to D. saltans. The data presented in this article support a model in which distribution of Minos transposon among Drosophila species is determined by horizontal transmission balanced by vertical inactivation and extinction.

P elements in the saltans group of Drosophila: a new evaluation of their distribution and number of genomic insertion sites

Few are studies on P elements that have addressed the saltans group. These studies had shown that species from the cordata and elliptica subgroups were devoid of any discernible P homologous sequences, while species from the parasaltans, sturtevanti, and saltans subgroups all contain P element sequences. Our analyses showed the presence of one to 15 P element insertion sites in species of the saltans group, including Drosophila neocordata and Drosophila emarginata ( cordata and elliptica subgroups, respectively). From these species, only those from the parasaltans, sturtevanti, and saltans subgroups harbor canonical P elements and, only those of the last two subgroups seem to harbor putative full-sized elements. Due to the low similarity of the sequences found in D. neocordata and D. emarginata to those earlier described, we suggest that these sequences might be rudimental P element derivatives that were present in the ancestral of the subgenus Sophophora.

Characterization of two full-sized P elements from Drosophila sturtevanti and Drosophila prosaltans

Previously, only partial P element sequences have been reported in the saltans group of Drosophila but in this paper we report two complete P element sequences from Drosophila sturtevanti and Drosophila prosaltans. The divergence of these sequences from the canonical P element of Drosophila melanogaster is about 31% at the nucleotide level. Phylogenetic analysis revealed that both elements belong to a clade of divergent sequences from the saltans and willistoni groups previously described by other authors.

Tree Rooting with Outgroups When They Differ in Their Nucleotide Composition from the Ingroup: The Drosophila saltans and willistoni Groups, a Case Study

Rooting is frequently the most precarious step in any phylogenetic analysis. Outgroups can become useless for rooting if they are too distantly related to the ingroup. Specifically, little attention has been paid to scenarios where outgroups have evolved different nucleotide frequencies from the ingroup. We investigate one empirical example that arose seeking to determine the phylogenetic relationship between the saltans and the willistoni groups of Drosophila (subgenus Sophophora). We have analyzed 2085 coding nucleotides from the xanthine dehydrogenase (Xdh) gene in 14 species, 6 from the saltans group and 8 from the willistoni group. We adopt a two-step strategy: (1) we investigate the phylogeny without outgroups, rooting the network by the midpoint method; (2) we reinvestigate the rooting of this phylogeny using predefined outgroups in both a parsimony- and a model-based maximum-likelihood framework. A satisfactory description of the substitution process along the Xdh region calls for six substitution types and substitution rate variation among codon positions. When the ingroup sequences are considered alone, the phylogeny obtained using this description corroborates the known relationships derived from anatomical criteria. Inclusion of the outgroups makes the root unstable, apparently because of differences between ingroups and outgroups in the substitution processes; these differences are better accounted for by a simplified model of evolution than by more complex, realistic descriptions of the substitution process.

Phylogenetic position of the subgenus Lordiphosa of the genus Drosophila (Diptera: Drosophilidae) inferred from alcohol dehydrogenase (Adh) gene sequences.

We analyzed the phylogenetic relationship between the species of Lordiphosa and other Drosophilidae using alcohol dehydrogenase (Adh) gene sequences. The phylogenetic trees consistently show that the four species Drosophila kurokawai, D. collinella, D. stackelbergi, and D. clarofinis, which include three species groups of Lordiphosa, form a monophyletic clade. This clade is placed as a sister group to the willistoni and saltans groups of Sophophora. On the other hand, three species of Lordiphosa, D. tenuicauda, D. pseudotenuicauda, and D. acutissima, all of which belong to the tenuicauda group, are not shown to be related to the major Lordiphosa lineage. In the phylogenetic trees, these species are included into the clade comprised of Drosophila and Hirtodrosophila, although it remains uncertain whether the tenuicauda group is a monophyletic group or not. These results indicate that Lordiphosa is polyphyletic and that most of the members of the subgenus have a close relationship to the neotropical groups of Sophophora. The above conclusion is compatible with the hypothesis of Okada (Mushi [1963] 37:79-100) and Lastovka and Máca (Acta Ent Bohemoslov [1978] 75:404-420) that Lordiphosa is most closely related to Sophophora; in contrast, our results contradict the hypothesis of Grimaldi (Bull Am Mus Nat Hist [1990] 197:1-139) that Lordiphosa is a sister group to the genus Scaptomyza.