Comparative copulation anatomy of the Drosophila melanogaster species complex (Diptera: Drosophilidae)

The Drosophila melanogaster species complex consists of four species: D. melanogaster, D. simulans, D. sechellia and D. mauritiana. To identify these closely related species, researchers often examine the male genitalia, especially species‐specific shapes of the posterior process, as the most reliable and easily observable character. However, compared to genetic aspects, the evolutionary significance of the posterior process and other genital parts remains largely unexplained. By comparing genital coupling among these species, we revealed that the posterior processes, which are hidden under the female abdominal tergite VII when genital coupling is established, mesh with different parts of the intersegmental membrane between the tergite VIII and the oviscapts and that this membrane region broadens in a species‐specific manner. Furthermore, in D. simulans and D. sechellia, this membrane region is likely to incur wounds from the sharply pointed tip of the posterior process. On the basis of the use and functions of these and other genital parts, we discuss possible evolutionary forces underlying the diversification of genitalia in this group.     

Fine structure of the sensory organs of Drosophila melanogaster Meigen larva (Diptera : Drosophilidae)

Fine structure of the prominent external and internal sensory organs of Drosophila melaogaster Meigan (Diptera : Drosophilidae) larva was determined by transmission electron microscopy (TEM). The external sensory organs, namely, antennal, maxillary, ventral and labial organs, dorsal pits, sensory cones on the 8th and 9th abdominal segments, and the sensory hairs on the body of the larva, were studied. A new knob in pit (KIP) sensillum innervated by 3 dendrites was found on the dorsolateral surface on either side. Four tufts of hairs at the posterior end of each great lateral tracheal trunk were found to be sensory with a dendrite at the base of each tuft.Internally in the pharynx of the larva, 3 groups of symmetrically located sensory organs, namely, the anteroventral, dorsal, and posteroventral groups were found. In all they contain 10 pairs of sensilla. Amongst them, 2 pairs of sensilla were found to be of the compound type; one having 9 dendrites arranged in 3 groups of 4, 3 and 2; while the other had 6 dendrites grouped as 2 and 4. In addition, 2 groups of sensilla were found on the internal dorsal fold on either side.Similarities were observed in the dendritic organisation of sensilla in the internal mouthparts of the Drosophila larva and the adult. Unlike nerves of the adult, the larval nerves connected with the dorsal and maxillary organs have a significantly thick layer of glial cells ensheathing the bundle of axons.     

Structure of cephalic abnormalities in Drosophila melanogaster (Diptera: Drosophilidae)

Scanning and transmission electron microscopy were utilized in studying the cephalic abnormalities of the tu-h strain of Drosophila melanogaster. Abnormalities, appearing as protuberances (growths) of different shapes and sizes, were observed only on or in close proximity to receptor cites. Compound eyes of some flies, besides having such protuberances, either had modified corneal lenses or were absent altogether. Cuticles of the growths were thinner than that of the normal surrounding layer. The epidermal cells associated with setae on the growths or underlying modified corneal lenses remained undifferntiated. Both setae and growths lacked innervation. The receptor portions (retinular sensory cells and secondary pigment cells) of the abnormal compound eye, where the dioptric portions were replaced by protuberances or remained undifferentiated, were unaffected by the mutation. Nuclei of several cells, including oenocytes and fat bodies, close to or underlying epidermal cells of abnormalities, were large and contained compact nucleoli without nucleonemas. Viruslike particles were observed in several nuclei of growth cells. It was concluded from the present study that the alterations induced by the mutation can be only a localized phenomenon restricted to at least several cephalic epidermal cells and/or their derivatives.     

A new trap and lure for Drosophila melanogaster (Diptera: Drosophilidae)

We conducted a series of nine laboratory experiments testing the response of "vinegar flies," Drosophila melanogaster Meigen (Diptera: Drosophilidae), released in bioassay chambers to experimental traps and lures. These experiments showed that an effective trap could be constructed from a clear 225-ml screw-cap jar fitted with a hollow 8-mm-diameter cylindrical cross bridge. Flies could enter the trap from either end of the cylindrical "gate" and in turn could enter the interior chamber of the trap through a cut out portion at mid-span of the cylinder. The experiments also showed that a natural-component lure could be made using a teabag containing freeze-dried banana powder, yeast, and carrageenan gum powder as a humectant. When dipped in water for 10-15 s and then placed in the bottom of a trap, the teabag provided effective attraction for at least 7 d. Captured flies were immobilized on a sticky card placed in the trap, allowing them to be easily seen. Unlike other traps that cannot be opened and have liquid lures, the cylindrical-gate trap can be reused repeatedly if the teabag and sticky card are replaced. A final two experiments showed that the prototype operational cylindrical-gate trap with a teabag lure captured 3.3 and 2.3 times more released flies, respectively, than the next best of three commercially available traps.     

Utilization of a Photosystem by Drosophila melanogaster Larvae (Diptera: Drosophilidae)

Foraging-stage third-instar larvae from most wild-type (normal) Drosophila melanogaster stocks are generally repelled by light. To identify factors that affect the larval photoresponse, we elucidated the effects of age, temperature, and time on the photoresponse of larvae from a wild-type Canton-S stock. In addition, we analyzed the larvae from the LI2 isofemale line, which are unresponsive to light in a photoassay. To determine whether LI2 larvae behave abnormally on other behavioral paradigms, in comparison to Canton-S controls, we tested larvae in taste and olfactory assays and observed them to determine whether they dispersed in a food source. Like Canton-S larvae, LI2 larvae and other isofemale lines whose progenitors were collected from the same natural population are responsive to taste and olfactory stimuli. Moreover, LI2 larvae disperse in the food source, as do Canton-S larvae tested in the dark. Larvae expressing para^sbl mutations, which respond normally to light but not to chemical stimuli, do not disperse normally in the food source, suggesting that dispersal may be mediated by perception of chemical cues.     

A genetic analysis of cyromazine resistance in Drosophila melanogaster (Diptera: Drosophilidae)

Flies resistant to the insect growth regulator cyromazine were selected in the F1 generation from a cyromazine-susceptible strain of Drosophila melanogaster (Meigen) treated with ethyl methanesulfonate. Four resistant strains were isolated by screening with cyromazine at a concentration > LC100 of susceptibles. In each strain, resistance is conferred by a single gene mutation. Cyromazine resistance in two of the mutants (rst(1a)cyr1 and rst(1a)cyr2) localizes to map position 17 of the X chromosome. Evidence is presented that these mutations are alleles of the gene rst(1a)cyr. Cyromazine resistance in another of the mutants (Rst(1b)Cyr) is also X-linked, and localizes to map position 49 of the X chromosome. The location of the gene conferring cyromazine resistance in the other mutant (Rst(2b)Cyr) is map position 66 of chromosome II. This is possibly an allele of a previously characterized cyromazine resistance gene, Rst(2)Cyr. Dosage-mortality analyses demonstrate a low level of cyromazine resistance is conferred in all strains.     

Isolation of dieldrin resistance from field populations of Drosophila melanogaster (Diptera: Drosophilidae)

High levels (about 4,000-fold) of resistance to dieldrin were isolated by screening field-collected populations of Drosophila melanogaster (Meigen). The resistance was made homozygous following 2-4 generations of selection. A single, major gene mapping to the left arm of chromosome III was solely responsible for resistance. The implications of the recovery of resistant mutants from field populations of D. melanogaster are discussed.     

Sensilla on the third antennal segment of Drosophila melanogaster meigen (Diptera : Drosophilidae)

The distribution and morphology of the sensilla on the 3rd antennal segment of Drosophila melanogaster Meigen (Diptera : Drosophilidae) were studied with light and electron microscopy. Four types of hairs were identified. Three types of hairs innervated by dendrites are sensilla basiconica, sensilla coeloconica and sensilla trichodea. They occur amongst a large number of the 4th type of uninnervated hairs or spinules.Sensilla basiconica and coeloconica can be easily identified by light microscopy on staining with 0.1016 silver nitrate in 70% ethanol. The tips of sensilla basiconica and coeloconica appear dark brown. Most of the sensilla trichodea and spinules remain unstained.Sensilla basiconica conform to the single-walled, multiporous sensilla, having poretubules and branched dendrites. Sensilla coeloconica are double-walled and have longitudinal channels near the tip. No wall pores are found on sensilla trichodea. Dendrites do not branch in sensilla coelonica and trichodea. A mechanosensory dendrite with characteristic tubular body is absent in these sensilla.Populations of sensilla basiconica and sensilla trichodea occur in diametrically opposite, distinct regions on the 3rd antennal segment-the former in the dorsomedial and the latter in the ventrolateral regions, whereas sensilla coeloconica are distributed on most of the anterior and posterior surfaces, including the cavity walls of the sacculus.The axons are arranged in distinct groups in the antennal nerves at the stalk of the 3rd segment. This grouping becomes more pronounced in the nerve prior to its entry into the brain.     

Phylogeny of a paradigm lineage: the Drosophila melanogaster species group (Diptera: Drosophilidae)

Although Drosophila melanogaster is a paradigm eukaryote for biology, relationships of this species and the other 174 species in the melanogaster species group are poorly explored and ambiguous. Gene regions of Cytochrome oxidase II (mt:CoII ), Alcohol dehydrogenase ( Adh ) and hunchback ( hb ) were sequenced and analysed phylogenetically to test prior hypotheses of relationships for the group based on chromosomes, morphology, and 28S rRNA gene sequences. A simultaneous cladistic analysis of the three newly sequenced gene regions produced a single well-resolved phylogeny for 49 exemplar species representing eight subgroups. Monophyly of each of the ananassae , melanogaster , montium , and takahashii subgroups is supported; the suzukii subgroup is polyphyletic. This phylogeny is consistent with variation in significant morphological structures, such as the male sex comb on the fore tarsus. The broad range of morphological variation among these species is interpreted and the applicability to evolution and developmental investigations is discussed. This phylogeny facilitates comparative investigations, such as gene family evolution, transposable element transmission, and evolution of morphological structures. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 76 , 21–37.     

Isolation of a deet-insensitive mutant of Drosophila melanogaster (Diptera: Drosophilidae).

Despite the widespread use of N,N,-diethyl-3-methylbenzamide (deet) in insect repellent products, nothing is known about the molecular basis for the repellency of deet, we initiated a molecular genetics program to elucidate the molecular mechanism of deet repellency in Drosophila melanogaster (Meigen). Deet repellency was apparently due to airborne vapors, as wild type flies were repelled by a deet-treated surface in the absence of physical contact and in the dark. A mutant was isolated using chemical mutagenesis and at choice assay. In a choice assay, mutant flies entered 82 +/- 1% of deet-containing tubes, whereas wild type flies entered only 6 +/- 2% of deet-containing tubes. The mutant was repelled by other repellents, benzaldehyde and citronellal. The mutation was recessive and located on the X chromosome.     

Genetic localization of DDT resistance in Drosophila melanogaster (Diptera: Drosophilidae).

Adult resistance to DDT conferred by various regions of the second chromosome of a DDT-resistant strain of Drosophila melanogaster (Meigen) was analyzed using the quantitative genetic method of Thoday and a topical test procedure. Resistance was localized in a small region near and to the left of cn. Both the genetic method and the topical test technique were effective. Results are consistent with the idea that the number of loci responsible for insecticide resistance in D. melanogaster is not large.     

Genetic basis of cross-resistance to three organophosphate insecticides in Drosophila melanogaster (Diptera: Drosophilidae)

To investigate the genetic basis of cross-resistance to insecticides, we conducted genetic analyses of resistance to three organophosphate insecticides, malathion, prothiophos, and fenitrothion. After isofemale lines resistant and susceptible to all of the three organophosphates had been screened from natural populations of Drosophila melanogaster (Meigen), chromosomal analyses were performed by using chromosome-substituted lines between the resistant and the susceptible lines. The chromosomal analyses revealed that both the second and the third chromosomes contributed to resistance to the organophosphates, suggesting that this resistant line possessed at least two factors for organophosphate resistance. However, the relative contribution of each chromosome was different in resistance to different organophosphates. We further carried out genetic mapping of a resistance factor for each organophosphate on each of the two chromosomes. Each resistance factor was mapped to the position of each chromosome, about II-62 and III-50. Results of the chromosomal analyses and the genetic mapping revealed that at least two resistance factors exhibiting different patterns of cross-resistance to the organophosphates existed within a natural population of D. melanogaster. Based on this research, genetic variation in insecticide resistance within natural populations and complex as well as simple aspects of the mechanism of cross-resistance are discussed.     

Anatomical and neurochemical classification of the antennal glomeruli in Drosophila melanogaster meigen (Diptera : Drosophilidae)

We are interested in how information about odours is processed in the antennal lobe of Drosophila melanogaster Meigen (Diptera : Drosophilidae). Our ultimate aim is to study how genes specify the development and functioning of the olfactory pathway. Such studies require a thorough knowledge of the anatomy, neurochemistry and development of the antennal system in normal files. This paper is a first attempt towards this end. Toluidine-methylene blue staining has revealed the presence of 22 glomeruli in the antennal lobe of Drosophila melanogaster. The number and size of glomeruli are invariant between individuals and the sexes. Fifteen of the glomeruli demonstrated specific uptake of choline, which is suggestive of some cholinergic innervation. In developmental studies, all glomeruli were found to be morphologically mature by 72 hr after formation of the white pupa. This study is a necessary first step towards an investigation of the role played by the glomeruli as functional units for odour coding.     

Mutagenic potential of permethrin in the Drosophila melanogaster (Diptera: Drosophilidae) sex-linked recessive lethal test

When permethrin was tested for mutagenicity in Drosophila melanogaster Meigen with the sex-linked recessive lethal test, it was nonmutagenic under conditions of this study. The frequencies of spontaneous mutation for permethrin and the negative control were 0.135% and 0.133%, respectively; the spontaneous mutation frequency for positive control was 12.6%. The difference between the mutation frequency of permethrin and the negative control was not significant.     

Fine structure of the spiracular glands in larval Drosophila melanogaster (Meig.) (Diptera : Drosophilidae)

The spiracular glands in the third-instar larvae of Drosophila melanogaster (Diptera : Drosophilidae) were examined with light and electron microscopy. Three club-shaped, unicellular glands are associated with each spiracle. Each gland has a large nucleus with a well-developed nucleolus and polytene chromosomes. Cytoplasmic features include a prominent plasma membrane reticular system (PMRS), which contains electron-dense material and gives rise to endocytotic vesicles. Numerous lipid droplets, mitochondria, free ribosomes, glycogen, lysosomes, and vesicles are scattered throughout the cytoplasm. The smooth endoplasmic reticulum is abundant, but rough endoplasmic reticulum and Golgi complexes are sparse. Small lipid droplets appear to fuse with one another to form larger droplets. In late third-instar glands, normal mitochondria decrease in number; they appear to degenerate and become converted into dense bodies with finely granular interiors. Several microvillous channels containing lipid are present within the cytoplasm. In the lumina of most of these channels are found the distal tips of cuticular ductules. The cuticular ductules merge with one another dorsally to form the main duct that carries the lipoid secretion to the spiracle cleft. The ultrastructure of the spiracular glands is consistent with their roles in the synthesis of lipoid secretion, which is used to provide hydrophobicity of the surface and to trap small particulate matter.     

The persistence of short‐term cold acclimation in Drosophila melanogaster (Diptera: Drosophilidae)

Daily and seasonal fluctuations in temperature present significant challenges for the survival of many ectothermic species that can be tempered via thermal acclimation. In the present study, we use multiple naturally derived genotypes of Drosophila melanogaster to determine the persistence of beneficial short‐term thermal acclimation on subsequent survival after cold shock. We found that the benefit of short‐term acclimation persisted for 2 h in most genotypes after a rapid cold hardening treatment. Genotype did not directly influence the persistence of short‐term acclimation benefits, indicating that environmental variation may be more important for the persistence of acclimation benefits rather than genetic capacity for acclimation. The present study extends the current understanding of the limits and importance of short‐term acclimation events, providing greater detail on the timing of the loss of short‐term acclimation benefits in a genetically variable natural population.     

Control of Penicillium roqueforti (Thom) infection in cultures of Drosophila melanogaster (Meigen) (Diptera: Drosophilidae)

Abstract  Microbial contamination of artificial insect food media can jeopardise the viability, productivity and survival of many insect cultures, including Drosophila melanogaster . Here we investigated and improved upon control methods for one common contaminant, Penicillium roqueforti . We found that the combined effect of methyl p -hydroxybenzoate (23.7 mM), propionic acid (67.5 mM) and sorbic acid (8.9 mM) (PS^NPS treatment) was the most effective of the four candidate treatments, at inhibiting the growth of P. roqueforti. PS^NPS treatment inhibited 100% of visible P. roqueforti growth for 21 days (a complete D. melanogaster life cycle) and thus reduced the risk of transmitting infection to the next generation. Although the PS^NPS treatment negatively affected the two D. melanogaster fitness components, survivorship (number of adults) and biomass (live weight), it did not prevent successful reproduction and is suitable for short-term treatment of P. roqueforti infections.     

Blood-eye barrier of the developing drosophila melanogaster (Diptera : Drosophilidae)

For the past quarter century, it has been known that a blood-eye barrier exists in adult insects. However, the life stage at which the barrier arises and the anatomical correlate of the barrier were not known. Compound eye development in Drosophila melanogaster (Diptera : Drosophilidae) is essentially complete at approximately 140 h after pupariation ; or about 20 h prior to eclosion. A search for a blood-eye barrier spanned late third-instar larvae, through late pupal life sampled at 40 and 140 h post-pupariation. No blood-eye barrier is present in the eye discs of last-instar larvae, based on the presence of lanthanum tracer among ommatidial cells and their processes. Pleated-sheet septate junctions, which link larval ommatidial cells, are not yet capable of totally blocking tracer from paracellular passage. The blood-eye barrier is constructed in the early phase (0–60 h) of pupal development in the wake of apoptosis and new cellular reorganization. In the developing compound eye, mature photoreceptor neurons must then be protected from the ionic vagaries of hemolymph to become electrophysically competent. First vestiges of a barrier with occluding septate junctions are seen in the 40-h old pupa, and by 140 h, the barrier is complete. The barrier prevails throughout adult life.