Yoshiko N. Tobari

A profile of Drosophila species' enzymes assayed by electrophoresis. I. Number of alleles, heterozygosities, and linkage disequilibrium in glucose-metabolizing systems and some other enzymes

Seven isolated large populations of Drosophila belonging to five different species were examined by starch gel electrophoresis for allozyme variation. Six to eleven enzyme loci in the glucose-metabolizing system (group I) and six to eight enzyme loci (group II) which were not directly involved in the above-mentioned system were assayed. The parameters estimated were the average number of alleles per locus, allele frequencies, proportions of polymorphic loci, and average heterozygosity per population for group I and group II loci. The major finding is that genetic variability measured by allozyme variations is much higher for group II than for group I enzymes in terms of every parameter in all the populations. This is consistent with the earlier findings in D. ananassae by Gillespie and Kojima (1968). Linkage disequilibrium, a measure of genome integration, was computed between an enzyme locus and an inversion segment of the same chromosome. The preliminary analysis of this aspect of the study indicates that no substantial linkage disequilibrium builds up between the chromosomal segments unless the pair of segments is less than 10 centimorgan units apart.

Polytene Chromosomal Maps of 11 Drosophila Species: The Order of Genomic Scaffolds Inferred From Genetic and Physical Maps

The sequencing of the 12 genomes of members of the genus Drosophila was taken as an opportunity to reevaluate the genetic and physical maps for 11 of the species, in part to aid in the mapping of assembled scaffolds. Here, we present an overview of the importance of cytogenetic maps to Drosophila biology and to the concepts of chromosomal evolution. Physical and genetic markers were used to anchor the genome assembly scaffolds to the polytene chromosomal maps for each species. In addition, a computational approach was used to anchor smaller scaffolds on the basis of the analysis of syntenic blocks. We present the chromosomal map data from each of the 11 sequenced non-Drosophila melanogaster species as a series of sections. Each section reviews the history of the polytene chromosome maps for each species, presents the new polytene chromosome maps, and anchors the genomic scaffolds to the cytological maps using genetic and physical markers. The mapping data agree with Mullers idea that the majority of Drosophila genes are syntenic. Despite the conservation of genes within homologous chromosome arms across species, the karyotypes of these species have changed through the fusion of chromosomal arms followed by subsequent rearrangement events.

Retrovirus-like features and site specific insertions of a transposable element, tom, in Drosophila ananassae

SummaryThe tom element, putatively associated with optic morphology (Om) mutations in Drosophila ananassae, was identified as a retrovirus-like transposable element. The tom element was found to terminate with 475 (or 474) base pair direct repeats which are identical in sequence to each other. Southern blot and heteroduplex analyses showed the tom element to have high homology to 297 and 17.6, two retrotransposons found in D. melanogaster. As in the cases of 297 and 17.6, tom includes nucleotide sequences coding for a presumptive protease and reverse transcriptase, similar in amino acid sequence to those of the Moloney murine leukaemia virus. At the tom insertion site of the sn9g locus, a host DNA sequence (T)ATAT was found to be duplicated on each side of the tom insertion and all other tom elements examined were also flanked by (T)ATAT. In each of six cases, the 5′ flanking host sequence was TATAT. These results indicate that the target sequence of the tom element may be TATAT and that the entire region or a part of this sequence was duplicated on insertion of the tom element.

Cytogenetic analysis of recombination in males of Drosophila ananassae

Cytogenetic studies of recombination in males of Drosophila ananassae were carried out by examining F1 males derived from the mating of marker females, b se; bri ru of the BS stock, with males of two wild strains, TNG and L8. The male recombination values in both sections b-se (chromosome 2) and bri-ru (chromosome 3) are high in TNG F1 but extremely low in L8 F1 We demonstrate the presence of chiasmata in TNG F1 males at a frequency capable of accounting for the observed recombination values. A unique series of “iso-site aberrations” was also observed in TNG F1 males. Because of a parallelism in the distribution pattern between the chiasmata and the isosite aberrations, we propose that recombination in males of D. ananassae is meiotic in origin and that the iso-site aberrations are related to chiasma formation.

Reproductive Isolation Among Geographical Populations of Drosophila Bipectinata Duda (Diptera, Drosophilidae) with Recognition of Three Subspecies

Among D. bipectinata Duda, 1923, three subspecies, bipectinata from Southeast Asia (SEA) and Okinawa (OKN), szentivanii stat. nov. from Papua New Guinea (PNG) (Mather & Dobzhansky, 1962) and pacificiae ssp. nov. from South Pacific Ocean (SPO), are recognized. The external morphology of the reproductive organs and the numbers of teeth per row in the sex combs are different between the three subspecies. Furthermore, the sterility of hybrid males between strains from the different regions confirms the subspecies status of each population from SEA, PNG and SPO, together with different gene arrangements in the geographical populations. Although males of the strains from OKN (Okinawa), the northernmost population, show significant differences in the number of teeth of sex combs from males of SEA (Southeast Asia) strains, hybrid males between them are fertile.

Evolution in the Drosophila ananassae species subgroup

Drosophila ananassae and its relatives have many advantages as a model of genetic differentiation and speciation. In this report, we examine evolutionary relationships in the ananassae species subgroup using a multi-locus molecular data set, karyotypes, meiotic chromosome configuration, chromosomal inversions, morphological traits, and patterns of reproductive isolation. We describe several new taxa that are the closest known relatives of D. ananassae. Analysis of Y-chromosomal and mitochondrial haplotypes, shared chromosome arrangements, pre-mating isolation, and hybrid male sterility suggests that these taxa represent a recent evolutionary radiation and may experience substantial gene flow. We discuss possible evolutionary histories of these species and give a formal description of one of them as D. parapallidosa Tobari sp. n. The comparative framework established by this study, combined with the recent sequencing of the D. ananassae genome, will facilitate future studies of reproductive isolation, phenotypic variation, and genome evolution in this lineage.

Genetic analyses of several Drosophila ananassae-complex species show a low-frequency major gene for parthenogenesis that maps to chromosome 2.

Parthenogenetic strains of several species have been found in the genus Drosophila. The mode of diploidization in the eggs of females has been found to be post-meiotic nuclear fusion. The genetic basis for this parthenogenesis is not understood but is believed to be under the control of a complex polygenic system. We found parthenogenetic females in an isofemale strain (LAE345) of D. pallidosa-like collected in 1981 at Lae, Papua New Guinea, and established a parthenogenetically reproducing strain. Parthenogenetic strains of D. ananassae and D. pallidosa collected at Taputimu, American Samoa had also been established by Futch (1972). D. ananassae, D. pallidosa and D. pallidosa-like are very closely related species belonging to the ananassae complex of the ananassae species subgroup of the melanogaster species group. Using these three species, we found that more than 80% of females from parthenogenetic strains produced progeny parthenogenetically and that inter-specific hybrid females also produced impaternate progeny. In the present report, we demonstrate that the mode of parthenogenesis of D. ananassae appears to be the post-meiotic nuclear doubling of a single meiotic product, and that a major gene responsible for the parthenogenesis maps to the left arm of the second chromosome of D. ananassae. We also suggest that the genetic basis for parthenogenesis capacity may be identical among the three closely related species. We discuss the function of the gene required for parthenogenesis and its significance for the evolutionary process.

Molecular and histological characterizations of the Om(2D) mutants in Drosophila ananassae.

SummaryA series of transposon-induced optic morphology (Om) mutants found in a hypermutable marker stock of Drosophila ananassae provides a useful system for analyzing the molecular mechanism of eye morphogenesis. In the present study, one of the 25 Om loci so far reported, Om(2D), has been subjected to histological and molecular analyses as a first step toward understanding the role of Om genes in eye morphogenesis. Histological abnormalities observed during eye morphogenesis of the mutant, i.e. cell death within the eye-antennal discs of third instar larvae, and loss of the lamina, disorganized ommatidia and atrophied optic lobes in adults, were all comparable to those reported with various eye morphology mutants of D. melanogaster. Approximately 25 kb of genomic DNA including the Om(2D) locus was cloned by tom tagging. Southern blot and cloning analyses of two alleles of the Om(2D) locus revealed that insertions of the tom element occurred at three sites within 359 bp; two tandemly arrayed toms sharing one long terminal repeat at the junction and an internally deleted tom were present 359 by apart from each other in Om(2D)63, while a single tom in reverse orientation was present within the 359 by in Om (2D)10a. Host DNA sequences at the three insertion sites were TATAT or AATAT, and ATAT was duplicated upon the tom insertion. Three spontaneous revertants and one induced extreme derivative of Om(2D)63 were obtained and characterized. A complete revertant lost all the preexisting tom elements. Two partial revertants lost one or two of the preexisting tom elements. In the extreme derivative, an additional insertion sequence was found within the two tandem tom elements. Northern blot analysis showed two transcription units in the Om (2D) region: one was on the centromere side of the tom insertion site and expressed a 3.2 kb major RNA and several minor RNAs; the other resided on the telomere side of the tom insertion and expressed a 1.5 kb RNA. Both 3.2 kb and 1.5 kb transcripts were expressed throughout development, but the former was more abundant in mutant embryos and the latter more highly expressed in mutant third instar larvae than in the corresponding stages in wild type.

Crossing over does occur in males of Drosophila ananassae from natural populations

Spontaneous crossing over in males of Drosophila ananassae has been well demonstrated using F(1) individuals from crosses between marker stocks and wild type strains. However, the question of its occurrence in males from natural populations remained open. Here we present the cytological evidence that crossing over does occur in males of D. ananassae from two Brazilian populations, sampled nearly 21 years apart, and in two recently sampled populations, one from Indonesia and one from Okinawa, Japan. Cytological analysis of meiosis in males collected from nature and in sons of females from the same population inseminated in nature revealed the presence of chiasmata, inversion chiasmata, and isosite chromosome breakages in the diplotene cells in all sampled populations. These data demonstrate that reciprocal and nonreciprocal exchanges and chromosome breakages, previously reported as related events of male crossing over, do occur at variable frequencies among males from natural populations.

Male recombination in Brazilian populations of Drosophila ananassae

With few exceptions, spontaneous crossing over does not normally occur in male Drosophila. Drosophila ananassae males show considerable amounts of crossing over. In wild males of D. ananassae from Asian (2008) and Brazilian populations (1986 and 2007) variable frequencies of meiotic crossing over, estimated from chiasmata counts, suggested the existence of factors controlling male crossing over in these populations. To corroborate for such prediction, we present data on spontaneous recombination in F1 males of D. ananassae heterozygous for chromosomes of the same Brazilian populations (1986) and marker chromosomes using three testers stocks. Mean recombination value was low, although high variability existed between individual frequencies. Recombination frequencies between lines in each tester stock were not significantly different, excepting when the 3ple-px and 3ple-cy testers were compared (p < 0.05). These two testers differ in respect to the regional distribution of crossovers. The occurrence of recombination in chromosomes 2 and 3 in F1 males tested with e(65) se; bri ru was not related, suggesting they are under independent genetic control. Our data are consistent with proposed genetic factors controlling male crossing over in the tester stocks and to the presence of enhancers and suppressors of male crossing over segregating in the Brazilian populations (1986).