Morphometric Analysis of Male Genitalia in Sibling Species of Drosophila virilis Sturt.

The structure of male genital organs in sibling species of the virilis group of Drosophila was examined using methods of multivariate statistics. The differences among these species were estimated using 33 indices and 2 angle parameters. The intraspecific and interspecific correlation structure of the examined characters and the order of their divergence were established. The key characters with respect to forming interspecific differences in the virilis species group were identified. Based on these results, the relative systematic positions of the sibling species are discussed as well as similarities and differences of the pattern of relationships among the species from that generally accepted for the virilis group.     

Genetics of esterases in Drosophila of the virilis group. I. Characteristics of esterase patterns in D. virilis,D. texana,D. littoralis,and their hybrids

Starch and polyacrylamide gel electrophoreses have detected six esterase fractions in Drosophila of the virilis group. These esterases have been characterized in detail using a series of substrates and inhibitors and also thermal treatment. Differences in esterase patterns have been found between D. virilis, D. texana, and D. litoralis as well as between D. virilis stocks. An interstock polymorphism for different esterase patterns has been established with respect to the electrophoretic mobilities of a number of esterase fractions. In rare instances, it has been observed within some D. virilis stocks, too. There is specificity in organ distribution of esterase fractions in Drosophila. Monogenic control of the electrophoretic mobilities of esterase-2 and esterase-4 has been demonstrated in D. virilis, and a dimer structure has been found in esterase-2. Genes controlling esterase-2 and esterase-4 are located on the second chromosome (209.3 for esterase-2 and 192.0 for esterase-4). In interstock and interspecific hybrids, esterases usually manifest codominance. In interstock hybrids, esterase-2 forms a hybrid band not observed in interspecific hybrids. In third instar larvae of interspecific hybrids, differential expression of certain esterase isozymes has been noted. These observations are in agreement with data from histochemical studies of organs of different hybrids.     

Nucleotide and repeat length variation at the nonA gene of the Drosophila virilis group species and its effects on male courtship song

Genes found to affect male courtship song characters in Drosophila melanogaster are good candidates when tracing genes responsible for species-specific songs in other Drosophila species. It has previously been shown that Thr–Gly repeat length variation at the period gene affects song traits in D. melanogaster, which gives the repetitive regions a special interest. In this work, we have characterised the patterns of nucleotide variation for gene regions containing two Gly and one Gln–Ala repeat in another D. melanogaster song gene, no-on-transient A, in D. virilis group species. The levels of nucleotide variability in D. virilis nonA were similar to those found for other genes of the species, and the gene sequences showed no signs of deviation from neutrality. The Gly 2 repeat preceding the central domain of the gene exhibited length variation, which did not, however, correlate with song variation either within D. virilis or between the species of D. virilis group. The Gly 3 repeat located on the other side of the central domain showed amino acid divergence parallel to the consensus phylogeny of the D. virilis group species. The species of the virilis subgroup having Asn after the first three glycines in this repeat have simple songs with no species-specificity, while the species of the montana subgroup having two Gly or Asn–Ser in this site have unique courtship songs. Amino acid differences between the species in this repeat may, however, reflect species phylogeny rather than have an effect on song divergence per se.     

Morphological analysis of male mating organ in the Drosophila virilis species group: a multivariate approach

The shape of the male mating organ differs among 11 closely related species of the Drosophila virilis species group. Multivariate analyses of variation of a suite of 35 morphological traits (indices) describing the phallus shape were carried out in order to characterize interspecies variability of the traits. An overwhelming majority of the traits displayed species‐specific variability. The main result of the investigation was the revelation of the differences involved in the traits studied in the evolution of the D. virilis species group. The structure of species‐specific variability of some traits was discovered to be in accordance with the generally accepted taxonomy of the species group, while that of other traits required isolation of D. virilisper se from lummei phylad (former virilis phylad), and confirmed separation of montana phylad into three subphylads: montana proper, littoralis and kanekoi. Several subsets of traits having separate variability were determined in different parts of the male mating organ which correspond to spots of evolutionarily significant variability.     

Variation of 3′-Terminal Fragment of 16S rRNA Gene in Closely Related Species of <Emphasis Type="Italic">Drosophila virilis</Emphasis> Group

Primary sequence of 3′-terminal fragment of the mitochondrial 16S rRNA gene has been determined in 12 Drosophila species of the virilis group. The functionally important elements in secondary structure of the RNA product were defined. The region corresponding to the peptidyltransferase center has been localized. Variation of the 3′-terminal region of 16S rRNA gene has been described in 12 species of the virilis group. Phylogeny of the Drosophila virilis species group is discussed.__________Translated from Genetika, Vol. 41, No. 8, 2005, pp. 1049–1054.Original Russian Text Copyright © 2005 by Sorokina, Mugue, Andrianov, Mitrofanov.     

THE GENETIC BASIS OF EVOLUTION OF THE MALE COURTSHIP SOUNDS IN THE DROSOPHILA VIRILIS GROUP

When courting, males of the Drosophila virilis group vibrate their wings and emit species-specific courtship sounds consisting of trains of polycyclic sound pulses. To analyze the genetic basis of evolutionary changes in the sounds we made an F₁ diallel set of reciprocal crosses between the members of the virilis phylad of the group (two stocks of D. virilis and one of D. americana americana, D. a. texana, D. novamexicana, and D. lummei). We also crossed the D. virilis stocks with the members of the montana phylad of the same group (D. kanekoi, D. littoralis, D. borealis, D. flavomontana, D. lacicola, and D. montana) and made a backcross (D. virilis x D. littoralis) x D. virilis using a D. virilis marker stock (b; sv t tb gp; cd; pe). The sounds of the hybrids were analyzed using the following parameters: the length of a pulse train (PTL), the number of pulses in a train (PN), the interpulse interval (IPI), the length of a pulse (PL), the number of cycles in a pulse (CN), and the length of a cycle (CL). In the virilis phylad, the differences between species appeared to be determined mainly by autosomal genes in each sound trait. The heritabilities (narrow-/broad-sense) obtained from the diallel tables were the following: PTL 0.662/0.817, PN 0.651/0.841, IPI 0.193/0.546, PL 0.408/0.552, CN 0.425/0.719, and CL 0.361/0.764. The direction of dominance is for longer PTL, higher PN and CN, and shorter IPI and CL. PL shows ambidirectional dominance. In the sounds of the virilis phylad species, PTL and PL seem to be phenotypically the most important parameters, since their components (PN and IPI for PTL, CN and CL for PL) are negatively correlated. In crosses between D. virilis and D. littoralis or D. flavomontana reciprocal hybrids differed from each other in PTL, IPI, PL, and CN indicating X-chromosomal or cytoplasmic inheritance. In the backcrosses between D. virilis and D. littoralis the role of the X chromosome was ascertained to be decisive. We conclude that an X-chromosomal major change allowing variation in IPI has occurred during the separation of the two D. virilis group phylads, the long IPI allowing variation also in PL (and CN). The evolution of the sounds in the virilis phylad has probably gone towards longer and denser pulse trains, while in the montana phylad the sounds have evolved in different directions.     

Nontranscribed spacers in Drosophila ribosomal DNA

Ribosomal DNA nontranscribed spacers in Drosophila virilis DNA have been examined in some detail by restriction site analysis of cloned segments of rDNA, nucleic acid hybridizations involving unfractionated rDNA, and base composition estimates. The overall G+C content of the spacer is 27–28%; this compares with 39% for rDNA as a whole, 40% for main band DNA, and 26% for the D. virilis satellites. Much of the spacer is comprised of 0.25 kb repeats revealed by digestion with Msp I, Fnu DII or Rsd I, which terminate very near the beginning of the template for the ribosomal RNA precursor. The spacers are heterogeneous in length among rDNA repeats, and this is largely accounted for by variation among rDNA units in the number of 0.25 kb elements per spacer. Despite its high A+T content and the repetitive nature of much of the spacer, and the proximity of rDNA and heterochromatin in Drosophila, pyrimidine tract analysis gave no indication of relatedness between the spacer and satellite DNA sequences. Species of Drosophila closely related to D. virilis have rDNA spacers that are homologous with those in D. virilis to the extent that hybridization of a cloned spacer segment of D. virilis rDNA to various DNA is comparable with hybridization to homologous DNA, and distributions of restriction enzyme cleavage sites are very similar (but not identical) among spacers of the various species. There is spacer length heterogeneity in the rDNA of all species, and each species has a unique major rDNA spacer length. Judging from Southern blot hybridization, D. hydei rDNA spacers have 20–30% sequence homology with D. virilis rDNA spacers, and a repetitive component is similarly sensitive to Msp I and Fnu DII digestion, D. melanogaster rDNA spacers have little or no homology with counterparts in D. virilis rDNA, despite a similar content of 0.25 kb repetitive elements. In contrast, sequences in rDNA that encode 18S and 28S ribosomal RNA have been highly conserved during the divergence of Drosophila species; this is inferred from interspecific hybridizations involving ribosomal RNA and a comparison of distributions of restriction enzyme cleavage sites in rDNA.Dedicated to Professor Wolfgang Beermann on the occasion of his sixtieth birthday     

Analysis of DNAs from two species of the virilis group of Drosophila and implications for satellite DNA evolution

The DNAs from two virilis group species of Drosophila, D. lummei and D. kanekoi, have been analyzed. D. lummei DNA has a major satellite which, on the basis of CsCl equilibrium centrifugation, thermal denaturation, renaturation and in situ hybridization is identical to D. virilis satellite I. D. kanekoi DNA has a major satellite at the same buoyant density in neutral CsCl gradients as satellite III of D. virilis. However, on the basis of alkaline CsCl gradients, the satellite contains a major and a minor component, neither one of which is identical to D. virilis satellite III. By in situ hybridization experiments, sequences complementary to the major component of the D. kanekoi satellite are detected in only some species and in a way not consistent with the phylogeny of the group. However, by filter hybridization experiments using nick-translated D. kanekoi satellite as well as D. lummei satellite I and D. virilis satellite III DNAs as probes, homologous sequences are detected in the DNAs of all virilis group species. Surprisingly, sequences homologous to these satellite DNAs are detected in DNAs from non-virilis group Drosophila species as well as from yeast, sea urchin, Xenopus and mouse.     

The pattern of polytene chromosome synapsis in Drosophila species and interspecific hybrids

The pairing of polytene chromosomes was investigated in Drosophila melanogaster, Drosophila simulans and their hybrids as well as in species of the D. virilis group and in F₁ hybrids between the species of this group. The study of frequency and extent of asynapsis revealed non-random distribution along chromosome arms both in interspecific hybrids and pure Drosophila species. It is suggested that definite chromosome regions exhibiting high pairing frequency serve as initiation sites of synapsis in salivary gland chromosomes.     

Variation of wing shape in the Drosophila virilis species group (Diptera: Drosophilidae)

Wing shape variation was analysed with geometric morphometric methods in 17 laboratory strains, representing 11 closely related species (including two subspecies) of the Drosophila virilis group: D. virilis, D. lummei, D. novamexicana, D. americana americana, D. americana texana, D. montana, D. lacicola, D. flavomontana, D. borealis, D. littoralis, D. ezoana and D. kanekoi. Overall shape estimated using Procrustes coordinates of 14 landmarks was highly variable among strains and very similar in females and males. The landmarks in the distal part of the wing showed higher variation across strains than those in the proximal part. Procrustes distances between species were not consistent with phylogenetic distances previously suggested for the virilis group. Moreover, Procrustes distances between strains within species and within two major phylads (virilis and montana) were comparable with those between species and between phylads, respectively. The most different from other members of the group was the endemic D. kanekoi species, currently viewed as separate subphylad within the montana phylad. Allometric effects were found to be partly responsible for shape differences between the strains. Three most significant shape transformations were considered using the relative warp analysis and the strains were ordinated in accordance with transformation values. The pattern of relative warp scores could be easily interpreted only for the third warp explaining about 13% of shape variation. It separated the largest species, D. montana, D. ezoana and D. kanekoi, from other ones and was mainly associated with shape changes in the proximal region of the wing. The results of the present work suggest that wing shape in the virilis species group is not related to the speciation process. The observed proximal‐distal contrasts and allometric effects are in agreement with data of other studies, in which wing shape variation was analysed within Drosophila species.     

Dominance status of shape of male genitalia in interspecific crosses of some Drosophila virilis group species

The heritability of the shape of the main species-specific morphological trait for the Drosophila virilis group-the male mating organ has been analyzed using the hybrid males D. virilis × D. lummei and D. virilis × D. novamexicana. The results suggest an increase in the share of the characters with a recessive status in the evolutionarily younger species and demonstrate the role of sex chromosomes in the implementation of a dominant or recessive status of the trait. The roles of additive and epistatic components of the total variation in the evolution of the dominance status, shown in several known theoretical models and confirmed by our data, are considered. The published data on sterility of hybrid males in interspecific crosses are discussed from the standpoint of the evolution of dominance.     

In Situ Hybridisation Analysis of the X-Linked Genes in the Species of the Virilis Group of Drosophila

When estimating the level of DNA sequence variation within and between populations or when planning QTL analysis, it is essential to know the location of the genes under study. In the present work, five X chromosomal genes, earlier localised in Drosophila virilis and D. littoralis, were mapped by in situ hybridisation on the larval polytene chromosomes of four other virilis group species, D. a. americana, D. flavomontana, D. lacicola and D. montana. Conjugation of X chromosomes of the most interesting species pairs was studied in interspecific hybrids. Three of the marker genes were used as RFLP markers to examine the occurrence of recombination in D. flavomontana and D. montana hybrid females. The gene arrangement of all species studied, appeared to be different at the proximal end of the X chromosome, which prevented normal conjugation along the most part of the X chromosome. The data illustrating the locations of five X chromosomal marker genes are presented for D. a. americana, D. flavomontana, D. lacicola and D. montana.     

Detection and location of three simple sequence DNAs in polytene chromosomes from virilis group species of Drosophila

In vitro synthesized RNAs complementary to the three satellite DNAs of Drosophila virilis have been used in a series of in situ hybridization experiments with polytene chromosomes from virilis group species. Gall and Atherton (1974) demonstrated that each of the satellites of D. virilis is comprised of many repeats of a distinct, seven base pair long, simple sequence. With few exceptions, copies of each of these simple sequences are detected in the chromocenters of all virilis group species. This is true even in species which do not possess satellite DNAs at buoyant densities corresponding to those of the satellite DNAs of D. virilis. Small quantities of the three simple sequences are also detected in euchromatic arms of several different species. The same euchromatic location may contain detectable copies of one, two, or all three simple sequence DNAs. The amounts of simple sequences at each location in the euchromatin may vary between species, between different stocks of the same species, and even between individuals of the same stock. The simple sequences located in the euchromatin appear to undergo DNA replication during formation of polytene chromosomes unlike those in heterochromatin. The locations of the euchromatic sequences are not the results of single chromosomal inversion events involving heterochromatic and euchromatic breakpoints.     

Histone content in relation to amount of heterochromatin and developmental stage in three species of Drosophila

Relative amounts of various histone fractions in Drosophila chromatin were estimated densitometrically on electrophoretic gel separations. Several consistent and highly significant differences were obtained between larval and adult chromatin. The arginine-rich histones showed the most conspicuous changes: higher amounts of H4 in larvae, higher H3 in adults. The level of modification of these histones was clearly higher in larval than in adult chromatin. The modification of the two slower subfractions of H4 involved, in all probability, phosphorylation as well as acetylation. In all types of Drosophila chromatin studied 50% or more of the H2a molecules were phosphorylated—a remarkably high proportion. The species differences observed in relative amounts of histone were consistent in both stages of development. D. melanogaster differed from D. hydei and D. virilis in all histones except H2b, while the latter two species were generally similar. The interspecific variation in histone pattern was generally not correlated to differences in content of heterochromatin. The level of modification of H3 was, however, presumably an exception, as it was significantly lower for both larvae and adults in D. virilis than in the other two species. These differ from D. virilis in containing appreciably lower proportions of heterochromatic chromosome segments.     

Distribution and evolution of mobile elements in the virilis species group of Drosophila

The distributions of Penelope and Ulysses, two transposable elements that can induce hybrid dysgenesis, were studied in several species groups of Drosophila. No significant hybridization to Penelope and Ulysses probes was detected by Southern blot analyses of species outside the virilis group. In contrast, both element families have had a long residence in all species of the virilis species group, as indicated by their strong presence in the heterochromatic chromocenter. Except for D. kanekoi, D. lummei, and some strains of D. virilis, species of the group carry full-sized, and at least potentially functional, copies of both element families. Consistent with the occurrence of recent transposition, Penelope and Ulysses elements are located at different chromosomal sites in different geographical strains of the same species. A total of 79 Penelope and 47 Ulysses euchromatic insertion sites were localized to chromosomal subsections in species of the virilis group. Highly significant deviations from independence of the distributions of Penelope and Ulysses and previously established inversion breakpoints were documented, suggesting that these transposable elements may have played an important role in genomic reorganization and evolution of the virilis species group, which is especially rich in karyotypic variation. Received: 13 April 1999; in revised form: 20 July 1999 / Accepted: 27 July 1999     

Evolution of the Response to Heat Shock in Genus Drosophila

Thermotolerance was studied in a wide spectrum of Drosophilaspecies and strains originating from different climatic zones and considerably differing from one another in the ambient temperature of their habitats. The species that lived in hot climate have a higher thermotolerance. Most species of the virilisgroup exhibited positive correlation between the HSP70 accumulation after heat exposure and thermotolerance; however, this correlation was absent in some species and strains. For example, the D. melanogasterOregon R strain, which had the highest sensitivity to heat shock (HS) among all strains and species studied, displayed the maximum level of HSP70 proteins after HS. The patterns of induction of various heat shock protein (HSP) families after heat exposure in a wide spectrum of Drosophila species were compared. The results obtained suggest that the HSP40 and low-molecular-weight HSPs (lmwHSPs) play a significant role in thermotolerance and adaptation to hot climate. Polymorphism in hsp70 gene clusters ofDrosophila and variation in the numbers of gene copies andhsp70 isoforms in group viriliswere found. The evolutionary role of the variation in the number of hsp70 gene copies observed in the strains and species of genusDrosophilais discussed.     

Isolation and characterization of microsatellites in Drosophila virilis and their cross species amplification in members of the D. virilis group

We report the isolation and cross species amplification of 42 Drosophila virilis microsatellite loci. Nine loci were isolated from mapped P1 bacteriophage clones and 33 were obtained from genomic DNA or GenBank searches. Cross species amplification was tested for all members of the D. virilis group. The amplification success was high (varying from 45% to 100%) and most of the loci were polymorphic. This set of loci can be applied for several genetic studies such as mapping behavioural quantitative trait loci (QTL) and for studying population structure in a phylogeographical framework in D. virilis group species.     

Invasion of an occupied niche by the crayfish Orconectes rusticus: potential importance of growth and mortality

We are exploring mechanisms of an invasion that contradicts the oft-cited generalization that species invade vacant niches. In northern Wisconsin lakes, the introduced crayfish Orconectes rusticus is replacing two ecologically similar resident congeners, O. virilis and O. propinquus. In laboratory experiments, we compared growth and mortality of individually maintained crayfish offered one of five ad libitum diets: invertebrates, macrophytes, dentritus, periphyton or all items combined. Mortality was highest for O. virilis and lowest for O. rusticus. Macrophyte diets yielded the highest mortality. All three species grew best on invertebrate and combination diets but grew little or not at all on diets of periphyton, detritus or macrophytes. O. rusticus and O. virilis grew more than O. propinquus. O. rusticus grew more quickly and/or was better able to survive overall than its congeners. Therefore, O. rusticus would probably have advantages over O. virilis and O. propinquus in competitive interactions, reproductive success and avoiding size-selective fish predation. Subtle interspecific differences may interact strongly with other ecological factors and contribute to the displacement of resident species from a well-occupied niche.