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The origin and possible time of divergence of the Hawaiian Drosophilidae: evidence from DNA sequences.

The origins and times of divergence of the speciose Hawaiian Drosophilidae are examined using mtDNA sequences. The Hawaiian Drosophilidae are resolved as the sister group to the subgenus Drosophila. No one member of the subgenus Drosophila could be determined to be more closely related to the Hawaiian Drosophilidae than could any other. This result suggests that the Hawaiian Drosophilidae arose before the diversification of the subgenus Drosophila and after the divergence of the subgenus Sophophora. In light of fossil evidence, this phylogenetic scenario suggests that the Hawaiian Drosophilidae lineages are no younger than 30 Myr.

Alcohol Dehydrogenase↗

Character congruence of multiple data partitions and the origin of the Hawaiian Drosophilidae.

We address two systematic questions, the origin and monophyly of the Hawaiian Drosophilidae and the question of how to analyze multiple data partitions containing disparate kinds of information, in this study. New evidence, from analysis of characters from morphology, mitochondrial DNA, and nuclear DNA suggests that the Hawaiian Drosophilidae are a monophyletic assemblage. Although we are not able to suggest a single species group which may represent the continental sister taxon to the Hawaiian Drosophilidae, the candidates suggested by our analysis, the melanicarobusta lineage and the virilis-repleta lineage, have been suggested by other investigators as probable sister taxa. Through analyses of our data partitions separately and in a variety of combinations, we conclude that the best phylogenetic hypothesis to be generated from multiple data partitions will be obtained if the data from these partitions are combined and analyzed simultaneously. We advocate performing tests for congruence between and among data partitions not to justify keeping them apart, but for understanding the interactions among the signals included in the analysis and how they affect the topology of, and support for, the total evidence tree.

Animals↗

Phylogeny of Drosophilinae (Diptera: Drosophilidae), with comments on combined analysis and character support.

Drosophilidae (Diptera) is a diverse, cosmopolitan family of flies. Here, we present a combined analysis phylogeny of Drosophilinae, one of the two subfamilies of Drosophilidae, based on data from six different data partitions, including both molecular and morphological characters. Although our data show support for the monophyly of the Hawaiian Drosophilidae, and the subgenus Sophophora, neither the genus Drosophila nor the subgenus Drosophila is monophyletic. Partitioned Bremer support (PBS) indicates that morphological data taken from Grimaldi's monograph (Grimaldi, 1990a), as well as sequences from the mitochondrial (mt) 16S rDNA and the nuclear Adh gene, lend much support to our tree's topology. This is particularly interesting in the case of Grimaldi's data, since his published hypothesis conflicts with ours in significant ways. Our combined analysis cladogram phylogeny reflects the catch-all designation that the name Drosophila has become, in that the cladogram does not support the monophyly of either the genus or subgenus Drosophila.

Animals↗

The mariner transposable element in the Drosophilidae family.

The distribution of the mariner transposable element among Drosophilidae species was investigated using three different techniques, i.e. squash blots, Southern blots and PCR amplification, using two sets of primers (one corresponding to the Inverted Terminal Repeats and the other to two conserved regions of the putative transposase). Our results and those of others show that the distribution of mariner is not uniform and does not follow the phylogeny of the host species. An analysis of geographical distribution, based on endemic species, shows that mariner is mainly present in Asia and Africa. At least two hypotheses may be proposed to explain the specific and geographical distributions of this element. Firstly, they may be the results of several horizontal transmissions between Drosophila species and/or between Drosophila species and one or several donor species outside the Drosophilidae family. Secondly, these particular distributions may correspond to the evolution of the mariner element from an ancestral copy which was present in the ancestor of the Drosophilidae family.

Animals↗

Shared nucleotide composition biases among species and their impact on phylogenetic reconstructions of the Drosophilidae.

Compositional changes are a major feature of genome evolution. Overlooking nucleotide composition differences among sequences can seriously mislead phylogenetic reconstructions. Large compositional variation exists among the members of the family Drosophilidae. Until now, however, base composition differences have been largely neglected in the formulations of the nucleotide substitution process used to reconstruct the phylogeny of this important group of species. The present study adopts a maximum-likelihood framework of phylogenetic inference in order to analyze five nuclear gene regions and shows that (1) the pattern of compositional variation in the Drosophilidae does not match the phylogeny of the species; (2) accounting for the heterogeneous GC content with Galtier and Gouy's nucleotide substitution model leads to a tree that differs in significant aspects from the tree inferred when the nucleotide composition differences are ignored, even though both phylogenetic hypotheses attain strong nodal support in the bootstrap analyses; and (3) the LogDet distance correction cannot completely overcome the distorting effects of the compositional variation that exists among the species of the Drosophilidae. Our analyses confidently place the Chymomyza genus as an outgroup closer than the genus Scaptodrosophila to the Drosophila genus and conclusively support the monophyly of the Sophophora subgenus.

Alcohol Dehydrogenase↗

Evolution of the AMP-forming acetyl-CoA synthetase gene in the Drosophilidae family.

Analysis of the AMP-forming ACS gene was performed in 12 species of the Drosophilidae family. Systematically four introns, aligned at the same positions, were detected, but none of them showed a position similar to those known for species outside the Drosophilidae family. The average length of introns varied from 63 to 75 bp but in two species Drosophila takahashii and D. kikkawai the length of the second intron was 343 and 210 bp, respectively. In coding regions, about 80% of the third codon positions were substituted while first and second positions showed, respectively, 14% and 6% substitutions. Interestingly, the divergence observed at the protein level between species was very low. The phylogenetic tree based on the DNA sequences of the exons was mainly in agreement with taxonomic classification and previous molecular phylogenies except for D. ananassae, which appeared more closely related to D. subobscura and D. funebris than to the species of the melanogaster group.

Acetate-CoA Ligase↗

[Mammalian carcinogen-induced warts tumor in Drosophilidae: a test sensitive to the blastomogenic action of chemical compounds].

A basically new system has been developed to screen carcinogens in Drosophilidae, which is based on somatic mutagenesis and recombination. The system may induce tumors in Drosophilidae, which are recorded in adult insects. The test uses recessive mutation in the suppressor gene of growth of warts (wts) tumors. The new system is sensitive to a wide spectrum of mutagenic carcinogens that are naturally encountered. The sensitivity of the system to polycyclic aromatic hydrocarbons and aromatic amides is higher than that of classical tests. Dominant p53 gene mutation that ceases mutagen-induced apoptosis has been shown to increase the incidence of wts tumors by many times. The magnitude of the effect depends on the rate of mutant p53 expression. The increasing effect of p53 mutation extends to both somatic recombination and point mutations and deletions at the wts locus.

Animals↗

The phylogenetic relationships of flies in the family Drosophilidae deduced from mtDNA sequences.

The phylogenetic relationships of several taxa from representative genera, subgenera, groups, and subgroups in the Drosophilidae were examined using sequences from a 905-bp mtDNA fragment. Conventional cloning and sequencing techniques were used to obtain nucleotide sequences. In addition, polymerase chain reaction primers were designed for the rapid amplification and sequencing of this region for the species examined in the Drosophilidae. Phylogenetic analysis was done by cladistic techniques. Because of the coding nature of the 905-bp mtDNA fragment, several separate analyses of these sequences were performed. The genera Scaptomyza and Hirtodrosophila occupy ancestral branching positions in the molecular phylogeny. The genera Chymomyza and Zaprionus have intermediate branching positions, while the subgenera Drosophila and Sophophora are in the most derived position in the molecular phylogeny. Within the subgenus Sophophora, there is little resolution using these sequences, while within the subgenus Drosophila, D. melanica, D. funebris, and D. pinicola form a clade in a derived part of the phylogeny, with D. robusta and D. immigrans branching in an intermediate position in the phylogeny. D. mercatorum, a member of the repleta species group, occupies an ancestral position in the molecular phylogeny.

Animals↗

Molecular evidence for parallel evolution of adaptive syndromes in fig-breeding Lissocephala (Drosophilidae).

Afrotropical Lissocephala (Drosophilidae) breed strictly in syconia (figs) of Ficus (Moraceae) and have accordingly evolved specific features including modified female and eggshell morphologies, ovipositing, larval foraging, and mating behaviors. These various traits may exist as two or three alternative states. Each species displays a specific suite of traits so closely coordinated with one another that alternative states of the overall suites of traits can be seen as "adaptive syndromes." Three clear-cut adaptive syndromes can be recognized while two taxonomic lineages (juncta and sanu species groups) are traditionally accepted on the basis of male terminalia. A crucial evolutionary question results from the consideration that ecological clusters and taxonomic groups have conflicting compositions: the three syndromes are found in the juncta group while two of them occur in the sanu group. To resolve this conflict, we present molecular data which provide a robust phylogeny: mitochondrial DNA (12S + 16S ribosomal DNA and cytochrome b) sequence data are in agreement with one another regardless of the algorithm used. All molecular data consistently support male terminalia dichotomy. Such a level of consistency unambiguously indicates that parallel evolution of adaptive syndromes occurred. Thus, homoplasy may affect morphological and behavioral traits concomitantly when these are involved in a network of functional relationships.

Adaptation, Physiological↗

Phylogenetic analysis of Mos1-like transposable elements in the Drosophilidae.

We have performed a phylogenetic analysis of 59 mariner elements in 14 Drosophilidae species that are related to the active Drosophila mauritiana Mos1 element. This includes 38 previously described sequences and 21 new sequences amplified by PCR from 10 species. Most of the elements detected are nonfunctional due to several frameshifts and deletions. They have been subdivided into four groups according to specific signatures in the nucleotidic and amino acid sequences. The mean nucleotide diversity is 4.8 +/- 0.1% and reflects mainly the divergence of inactive elements over different periods. Although this probably gives rise to occasional homoplasies between distantly related taxa, the elements of each species remain grouped together. Horizontal transfer, reported previously between D. mauritiana and Zaprionus tuberculatus, can be extended to Z. verruca, while the Mos1-like element of Z. indianus belongs to another group. Interpretation of the phylogeny leads to a comparison of the influence of common ancestral sequences and putative horizontal transfers.

Animals↗

Do deletions of Mos1-like elements occur randomly in the Drosophilidae family?

We compared deleted copies of the seven mauritiana subfamilies of mariner transposable elements in species of the Drosophilidae. All elements were detected by PCR using the inverted terminal repeats of the Mos1 element of Drosophila mauritiana as primers. A higher frequency of breakpoints in the 5prime prime or minute part of the element compared to the 3prime prime or minute part was observed. Of the 27 deletions, 9 (33%) occurred between short direct repeats (SDR) of 5 to 8 bp. The SDRs can be at or close to the breakpoints of the deletion. A deleted copy of D. simulans (St. Martin population) had three repeats of a motif present only once in the complete consensus sequence. The high frequency of SDRs at or near the breakpoints of the deletions strongly suggests that some of them do not occur at random. Mechanisms that might explain these deletions, such as unequal crossing-over, ectopic recombination, and abortive gap repair, are discussed.

Animals↗

Variability of wing size and shape in three populations of a recent Brazilian invader, Zaprionus indianus (Diptera: Drosophilidae), from different habitats.

Zaprionus indianus (Diptera: Drosophilidae) is an African species that was introduced in Brazil near the end of the 1990's decade. To evaluate the adaptive potential of morphological traits in natural populations of this recently introduced species, we have investigated wing size and shape variation at Rio de Janeiro populations only two years after the first record of Z. indianus in Brazil. Significant genetic differences among populations from three distinct ecological habitats were detected. The heritability and evolvability estimates show that, even with the population bottleneck that should have occurred during the invasion event, an appreciable amount of additive genetic variation for wing size and shape was retained. Our results also indicated a greater influence of environmental variation on wing size than on wing shape. The importance of quantitative genetic variability and plasticity in the successful establishment and dispersal of Z. indianus in the Brazilian territory is then discussed.

Animals↗

The evolution of HOM-C homeoboxes in the Dipteran family Drosophilidae.

Forty-five new Homeotic Complex (HOM-C) homeobox sequences from six species of Drosophilidae (Drosophila heteroneura, D. adiastola, Zaprionus vittiger, Chymomyza amoena, Scaptodrosophila pattersoni and Hirtodrosophila pictiventris) were obtained using a PCR-cloning method. These new homeoboxes are from the labial, proboscipedia, Deformed, Sex combs reduced, fushi tarazu, Antennapedia, Ultrabithorax, abdominal-A and Abdominal-B genes. Phylogenetic signal in the homeobox sequences was assessed and several aspects of sequence evolution were examined. In particular, codon bias was examined and found to exist between the drosophilid species examined here and Anopheles gambiae outgroup sequences. In addition, different patterns of codon bias were detected in homeoboxes interrupted with introns when compared to homeoboxes that are uninterrupted.

Amino Acid Sequence↗

Shallow genomics, phylogenetics, and evolution in the family Drosophilidae.

The effects of the genomic revolution are beginning to be felt in all disciplines of the biological sciences. Evolutionary biology in general, and phylogenetic systematics in particular, are being revolutionized by these advances. The advent of rapid nucleotide sequencing techniques have provided phylogenetic biologists with the tools required to quickly and efficiently generate large amounts of character information. We use family Drosophilidae as a model system to study phylogenetics and genome evolution by combining high throughput sequencing methods from the field genomics and standard phylogenetic methodology. This paper presents preliminary results from this work. Separate data partitions, based on either gene function or linkage group, are compared to a combined analysis of all the data to assess support on phylogenetic trees.

Animals↗

Two or four bristles: functional evolution of an enhancer of scute in Drosophilidae.

Changes in cis-regulatory sequences are proposed to underlie much of morphological evolution. Yet, little is known about how such modifications translate into phenotypic differences. To address this problem, we focus on the dorsocentral bristles of Drosophilidae. In Drosophila melanogaster, development of these bristles depends on a cis-regulatory element, the dorsocentral enhancer, to activate scute in a cluster of cells from which two bristles on the posterior scutum arise. A few species however, such as D. quadrilineata, bear anterior dorsocentral bristles as well as posterior ones, a derived feature. This correlates with an anterior expansion of the scute expression domain. Here, we show that the D. quadrilineata enhancer has evolved, and is now active in more anterior regions. When used to rescue scute expression in transgenic D. melanogaster, the D. quadrilineata enhancer is able to induce anterior bristles. Importantly, these properties are not displayed by homologous enhancers from control species bearing only two posterior bristles. We also provide evidence that upstream regulation of the enhancer, by the GATA transcription factor Pannier, has been evolutionarily conserved. This work illustrates how, in the context of a conserved trans-regulatory landscape, evolutionary tinkering of pre-existing enhancers can modify gene expression patterns and contribute to morphological diversification.

Animals↗

Species to genera: phylogenetic inference in the Hawaiian Drosophilidae.

Systematic studies at any taxonomic level require careful planning, even before genes are sequenced or morphological characters scored. Molecular systematists working at the level between species and genera must select gene regions which are suitable for the divergence within the group being examined and decide how many and which ingroup and outgroup taxa to sample in the analysis. This chapter will use studies of the Hawaiian Drosophilidae to illustrate strategies useful in (1) selecting nucleotide sequences for divergences between the levels of species and genera, (2) designing ingroup and outgroup taxon sampling schemes, and (3) performing phylogenetic analysis on data sets with large numbers of taxa and characters.

Animals↗

The formation of the functional chorion structure of Drosophila virilis involves inercalation of the "middle" and "late" major chorion proteins: a general model for chorion assembly in Drosophilidae.

We have shown by in vitro development of staged follicles in the presence of tritiated proline, as well as by immunoblot analysis, that the Dvs19 and DVS15 chorion proteins of Drosophila virilis are synthesized by the follicular epithelium and are incorporated in the chorion at the terminal stages of choriogenesis. Light microscopy immunolocalization has revealed that these two proteins represent structural components of the endochorion in all the specialized regions of the eggshell, while, contrary to their temporal pattern of secretion, electron microscopic immunolocalization has shown that these two proteins participate in endochorionic structures formed before the onset of their synthesis, thus suggesting an intercalation process during chorion formation. Furthermore, double immunolocalizations demonstrated that cosecreted proteins do not coexist in all secretory vesicles of the follicle cells, indicating that probably the final association of these chorionic components occurs extracellularly. On the basis of these data and those presented in the accompanying paper [Trougakos, I.P., and Margaritis, L.H. (1998) Immunolocalization of the temporally "early" secreted major structural chorion proteins, Dvs38 and Dvs36, in the eggshell layers and regions of Drosophila virilis, J. Struct. Biol. 123, 000-000.], a model for chorion assembly in Drosophilidae is proposed.

Animals↗

Mass determination of the unit cell of the innermost chorionic layer in Drosophilidae by scanning transmission electron microscopy.

The innermost chorionic layer (ICL) in eggshells of Drosophila melanogaster is a naturally occurring patchwork of thin three-dimensional crystalline plates located between the inner endochorion and the vitelline envelope. The mass-per-unit area of the ICL has been measured from scanning transmission electron microscope images of isolated unstained material and it was possible to distinguish up to four layers with the majority of the crystalline sheets being one to three layers thick. Taking into account the unit cell areas for the different crystals, we have estimated the mean ICL subunit sizes to be 36 kDa for Drosophila melanogaster, 35 kDa for Drosophila auraria, and 33 kDa for Drosophila teissieri. The results suggest that the three different Drosophilidae species have very similar average subunit masses.

Animals↗