Infection densities of three Spiroplasma strains in the host Drosophila melanogaster

Maternally transmitted endosymbiotic bacteria of the genus Spiroplasma associate with numerous insect species, including the genus Drosophila. Among the Spiroplasma strains associated with Drosophila, several manipulate their host??s reproduction by killing the male offspring of the infected females. Although the male-killing mechanism is not well understood, previous studies of non-native strains transferred to D. melanogaster (strain Oregon-R) indicate that the male-killing strain achieves higher densities than two non-male-killing strains. Whether this pattern of higher male-killing strain densities occurs in other host-symbiont strain combinations is not known. Herein, we used quantitative PCR to examine infection densities of one non-male-killing strain native to D. hydei (Hyd1), and two male-killing strains; one native to D. nebulosa (NSRO), and one native to D. melanogaster (MSRO; recently discovered), upon artificial transfer to D. melanogaster (strain Canton-S). Infection densities were examined at four weekly intervals in adult flies, across three consecutive generations following artificial transfer. Infection densities of the non-male-killing strain were significantly lower than those of the two male killers immediately after adult emergence. At later time points, however, the non-male-killing strain (Hyd1) is capable of proliferating to densities similar to those of the two male-killing strains (NSRO and MSRO) in D. melanogaster (Canton-S). We also examined the effect of co-infection by the heritable bacterium Wolbachia, on Spiroplasma densities and male-killing ability. Wolbachia had little to no effect of Spiroplasma densities, but the male-killing ability of MSRO was lower in the presence of Wolbachia. Generation post-infection had little effect on Spiroplasma densities, but affected the male-killing ability.     

Frequency-dependent viability in mutant strains of Drosophila melanogaster

We investigated the effects of genotypic frequencies on egg-to-adult viabilities in pairwise combinations of four strains of Drosophila melanogaster. The experiments involved mixture of a total of 42,000 eggs in varying proportions under controlled densities and observation of surviving adults. Viabilities were found to depend on frequencies in several genotypic combinations. In the most extreme case, the absolute viability of cn;bw females increased monotonically from 54% when common to 70% when rare. The results illustrate several statistical and methodological problems that might explain why some experiments have failed to detect frequency-dependent viabilities. These problems include heterogeneity between replications, sex differences in susceptibility to competition, and strong dependence of the experimental outcome on the choice of competitor genotypes.     

Variation in resistance and acclimation to low-temperature stress among three geographical strains of Drosophila melanogaster

Variation in cold resistance was examined in cold acclimated and non-acclimated Drosophila melanogaster from three geographical strains representing Morocco, France and Finland. Resistance was estimated as survival of adults at 0°C; the acclimation treatment involved a long-term exposure to 11°C starting from the late pupal stage and continuing with adults. After the cold stress, two fitness traits, percentage of fertile individuals and the number of adult progeny, were scored in both acclimated and non-acclimated flies. Acclimation dramatically increased survival in all strains, but did not affect the pattern of geographic variation in cold resistance. The European flies tended to be more resistant than the African ones and the ranking from most to least resistant strain was France>Finland>Morocco. In the absence of acclimation, females showed a higher survival than males. Percentage of fertile males in all strains and the number of progeny in the Finnish and French strains were decreased after acclimation. Without cold acclimation, the number of progeny was higher in the European flies as compared with the African ones. The results suggest that populations of D. melanogaster from cold climates are better adapted to low stressful temperatures and among-population variation in cold resistance may be due to non-plastic rather than plastic genetic changes. The deleterious effects of cold pretreatment on the life-history parameters indicate a possibility for acclimation costs in reproduction.     

Functional significance of seminal receptacle length in Drosophila melanogaster

Despite its central role in post-copulatory sexual selection, the female reproductive tract is poorly understood. Here we provide the first experimental study of the adaptive significance of variation in female sperm-storage organ morphology. Using populations of Drosophila melanogaster artificially selected for longer or shorter seminal receptacles, we identify relationships between the length of this primary sperm-storage organ and the number of sperm stored, pattern of progeny production, rate of egg fertilization, remating interval, and pattern of sperm precedence. Costs and benefits of relatively short or long organs were identified. Benefits of longer receptacles include increased sperm-storage capacity and thus progeny production from a single insemination. Results suggest that longer receptacles have not naturally evolved because of developmental time costs and a correlated reduction in longevity of mated females. This latter cost may be a consequence of sexual conflict mediated by ejaculate toxicity. Receptacle length did not alter the pattern of sperm precedence, which is consistent with data on the co-evolution of sperm and female receptacle length, and a pattern of differential male fertilization success being principally determined by the interaction between these male and female traits.     

Mutational variability of enzyme loci in unstable Drosophila melanogaster strains

Biochemical analysis of a number of related unstable Drosophila melanogaster strains was carried out. These strains have been shown to undergo mutational transformations caused by insertion/excision of transposable elements (Gerasimova, 1985). Activity and mobility variants of alpha-GPDH, ADH, SOD, G6PD, 6PGD and EST-6 were analysed. Two loci controlling SOD and 6PGD proved to be invariable, the loci alpha-Gpdh and Est-6 causing reduced activity of their products in the initial strain ctMR2. The direct and reversed transformations analogous to the mutational passages of morphological characters were traced in two loci controlling ADH and G6PD. The data obtained are discussed in terms of up-to-date views on the functional role of transposable elements, in relation to the genetic stability of the strains studied, under the multiple transpositions of mobile elements.     

Diflubenzuron inhibits reproduction of different strains of Drosophila melanogaster

Abstract  The effect of (1-(4-chlorophenyl)-3-(2,6-difluorobenzoyl)urea) insecticide – diflubenzuron – on wild type and white type fruit flies Drosophila melanogaster (Meigen, 1830) was studied. Adult insects were placed in vials with different concentrations of the insecticide in the nutrient (from 0.048 to 48 mg of diflubenzuron per 1 cm³ of the nutrient). In each case, the insects showed some mortality, which was concentration-dependant. When comparing both strains, we could observe different acute toxicities, with wild type being more resistant to diflubenzuron. However, subacute toxicity was similar within both strains. In both strains the prolonged exposure led to the extinction of the majority of the populations (>70%). Although imagoes could freely copulate, we did not observe copulating fruit flies, and we did not find either eggs or larvae in any of the exposed groups. These results indicate that genetic mutation which leads to different eye colour can also affect resistance and survival of insects in pesticide-exposed areas.     

Radioinduced change in life span of laboratory strains of Drosophila melanogaster

Chronic irradiation (accumulated dose 0.6-0.8 Gy) was shown to change the life span in male Drosophila melanogaster. Death was retarded in wild-type strains and accelerated in mutant strains defective in DNA repair and displaying a higher sensitivity to induction of apoptosis.     

Morpho-functional changes of fat body in bacteria fed Drosophila melanogaster strains

We have examined the addition of Escherichia coli to the diet at day 0 of adult life of females from two Oregon R Drosophila melanogaster strains, selected for different longevities: a short-life with an average adult life span of 10 days and a long-life standard R strain with an average adult life span of 50 days. The addition of bacteria to the diet significantly prolonged the fly longevity in both strains and affected the structure and histochemical reactivity of the fat body. The increased survival was characterized by great amount of glycogen accumulated in fat body cells from both strains. In aged control animals, fed with standard diet, lipid droplets were seen to be stored in fat body of short-lived, but not long-lived, flies. On the whole, our data indicate that exogenous bacteria are able to extend the survival of Drosophila females, and suggest that such a beneficial effect can be mediated, at least in part, by the fat body cells that likely play a role in modulating the accumulation and mobilization of reserve stores to ensure lifelong energy homeostasis.     

Retrotransposon gtwin: structural analysis and distribution in Drosophila melanogaster strains

A search for noncanonical variants of the gypsy retrotransposon (MDG4) in the genome of the Drosophila melanogaster strain G32 led to the cloning of four copies of the poorly studied 7411-bp gtwin element. Sequence analysis showed that gtwin belongs to a family of endogeneous retroviruses, which are widespread in the Drosophila genome and have recently been termed insect erantiviruses. The gtwin retrotransposon is evolutionarily closest to MDG4, as evident from a good alignment of their nucleotide sequences including ORF1 (the pol gene) and ORF3 (the env gene), as well as the amino acid sequences of their protein products. These regions showed more than 75% homology. The distribution of gtwin was studied in several strains of the genus Drosophila. While strain G32 contained more than 20 copies of the element, ten other D. melanogaster strains carried gtwin in two to six copies per genome. The gtwin element was not detected in D. hydei or D. virilis. Comparison of the cloned gtwin sequences with the gtwin sequence available from the D. melanogaster genome database showed that the two variants of the mobile element differ by the presence or absence of a stop codon in the central region of ORF3. Its absence from the gtwin copies cloned from the strain G32 may indicate an association between the functional state of ORF3 and amplification of the element.     

Variation in fertility of two wild type strains of Drosophila melanogaster meigen

Initial observations of low fertility, reduced sexual activity of males, and a high frequency of abnormalities in sperm differentiation of the wild type strain (Sevelen, Zürich) of Drosophila melanogaster, normally used in this laboratory, have lead to a study of this phenomenon and its causes. The abnormalities occur during all spermiogenetic stages and are not unique but are found with much lower frequency in normally fertile flies (Oregon R). Growth of Sevelen flies at high but sublethal temperatures (30°C) results in complete sterility, highly abnormal sperm differentiation, and a failure to recover fertility after return to normal temperatures (25° C) in the time period in which normal flies recover. — The principal factor, or factors, controlling normal sperm differentiation are located on the Y chromosome, but are thus far not precisely localized. Expression of the phenotype is modified by genetic background in which the Y chromosome is found.     

Microcompartmentation of cAMP in wild-type and memory-mutant dunce strains of Drosophila melanogaster

The "enzyme-probe" method [Solti M, Friedrich P: Eur J Biochem 95:551, 1979] has been applied to characterize the cyclic AMP pool in wild-type Canton-S and memory-mutant dunceM11 strains of Drosophila melanogaster. The kinetics of cyclic AMP breakdown in whole fly homogenates by endogenous cyclic nucleotide phosphodiesterase(s) indicate that the cyclic AMP pool is divided into free and bound fractions. The bound fraction in Canton-S and dunceM11 is 0.5 and 1.5 pmole/mg fly, respectively. Considering the total cyclic AMP content of the two strains, 1.6 and 10 pmole/mg fly, respectively, we conclude that the bulk of excess cyclic AMP in the mutant is free nucleotide.     

Structural analysis of the three vitellogenin genes in Drosophila melanogaster.

Genomic fragments coding for sequences expressed as abundant mRNA in female Drosophila melanogaster were isolated from a lambda library. Hybridization of these clones to polytene chromosomes. in situ, identified four which mapped to X chromosomal region 9A to 9B, the locus for yolk proteins 1 and 2 (Ypl,2) and two which mapped to 12A6-7 to 12D3, the locus for Yp3. These clones were mapped with restriction enzymes, and the coding regions and regions of homology determined by Southern blots probed with cDNA, 5'-end-labelled RNA and nick-translated DNA. Heteroduplex and R-loop mapping confirmed that three of the clones carried two genes separated by about 1.4 kb and oriented in opposite directions. Southern blots probed with cDNA made from alkali-hydrolyzed RNA showed that these genes had their 5' ends next to each other. All 3 genes show homology to each other and have a main coding region of about 1.3 kb, the approximate size for the mRNAs.     

Mating behavior in mutant strains of Drosophila melanogaster at different population densities

The effects of mutations and genetic background on the mating activity of males and receptivity of females Drosophila melanogaster have been studied at different population densities. Population density, as well as its combinations with other factors, significantly affects mating behavior of D. melanogaster. There are two distinct trends in the effect of this factor on mating behavior: the maximum larval overpopulation may cause either a significant suppression of the behaviors studied or an increase in their expressivity. The mating behaviors of w ^a and cn mutants against a certain genetic background changed similarly in response to varying population density.     

Mating behavior in mutant strains of Drosophila melanogaster at different population densities

The effects of mutations and genetic background on the mating activity of male and receptivity of female Drosophila melanogaster have been studied at different population densities. Population density, as well as its combinations with other factors, significantly affects mating behavior of D. melanogaster. There are two distinct trends in the effect of this factor on mating behavior: the maximum larval overpopulation may cause either a significant suppression of the behaviors studied or an increase in their expressivity. The mating behaviors of wa and cn mutants against a certain genetic background changed similarly in response to varying population density.     

Transposition bursts in analysis of repeated mutations in unstable strains of Drosophila melanogaster

The phenomenon of transposition memory was earlier demonstrated for the cut locus and mdg4. This work has been aimed at finding out, in what way the transposition memory can be realized. An unstable stock cmMR17ctMRpN17 was analysed which had high frequency of double cm+ct+ reversions and cmMRctMRpN repeated mutations. A series of five such transpositions could be followed. The ctMRpN17 mutation is a result of insertion at the cut locus mdg4 with the jockey element inserted within it. As seen from in situ hybridization analysis, transitions to the normal phenotype correlate, as a rule, with the excision of mdg4 and the jockey from the cut locus. Analysis of distribution of mdg1, mdg2, mdg3 and jockey on the X-chromosome of unstable revertants and repeated mutants indicated that not only transpositions of mdg4 and jockey, but also those of all mobile elements tested occur. So, we propose that the transposition memory in our genetic system is manifested in the process of transposition bursts.