Testing the Drosophila maternal haploid gene for functional divergence and a role in hybrid incompatibility

Crosses between Drosophila simulans females and Drosophila melanogaster males produce viable F1 sons and poorly viable F1 daughters. Unlike most hybrid incompatibilities, this hybrid incompatibility violates Haldane’s rule, the observation that incompatibilities preferentially affect the heterogametic sex. Furthermore, it has a different genetic basis than hybrid lethality in the reciprocal cross, with the causal allele in Drosophila melanogaster being a large species-specific block of complex satellite DNA on its X chromosome known as the 359-bp satellite, rather than a protein-coding locus. The causal allele(s) in Drosophila simulans are unknown but likely involve maternally expressed genes or factors since the F1 females die during early embryogenesis. The maternal haploid (mh) gene is an intriguing candidate because it is expressed maternally and its protein product localizes to the 359-bp repeat. We found that this gene has diverged extensively between Drosophila melanogaster and Drosophila simulans. This observation led to the hypothesis that Drosophila melanogaster mh may have coevolved with the 359-bp repeat and that hybrid incompatibility thus results from the absence of a coevolved mh allele in Drosophila simulans. We tested for the functional divergence of mh by creating matched transformants of Drosophila melanogaster and Drosophila simulans orthologs in both Drosophila melanogaster and Drosophila simulans strains. Surprisingly, we find that Drosophila simulans mh fully complements the female sterile phenotype of Drosophila melanogaster mh mutations. Contrary to our hypothesis, we find no evidence that adding a Drosophila melanogaster mh gene to Drosophila simulans increases hybrid viability

Nejire/dCBP-mediated control of H3 acetylation and transcriptional regulation by testis-specific Plus3 domain proteins during Drosophila spermatogenesis

Spermatogenesis describes the development from germ line stem cells to highly specialized sperm. Drosophila melanogaster spermatogenesis is a good model system for chromatin remodelling processes as many of these processes are similar in mammals and in flies. Histone modifications are a prerequisite for the exchange of histones by protamines during these chromatin remodelling processes but also transcription processes in earlier germ cell stages can be compared. In this thesis, the histone acetyltransferase Nejire/dCBP has been characterised as being responsible for the modification of histone H3 at lysine 18 and lysine 27 during post-meiotic germ cell development as well as in the spermatocyte stage. An RNAi-mediated knock-down revealed that the function of Nejire/dCBP is essential for fertility of male flies. Efficient mRNA synthesis of postmeiotic chromatin components depends on Nejire/dCBP whereas incorporation of protamines into the chromatin does not seem to depend on Nejire/dCBP function. Drosophila spermatogenesis is featured by a special regulation of transcription and translation. While most transcripts are synthesised in spermatocytes, a large portion has to be translationally repressed until required in later stages of germ cell development. Transcriptional regulation is supported by testis-specific variants of the general transcription machinery. This includes among others the tTAFs, the tMAC complex and bromodomain proteins. Bromodomain proteins are able to recognise and bind acetylated lysine residues on N-terminal histone chains. The bromodomain proteins tBRD-1 and tBRD-2 are expressed in spermatocytes and can interact with tTAFs, this might facilitate recruitment of the TFIID complex to certain chromatin areas. Further testis-specific variants of ubiquitously expressed proteins are the Plus3 domain proteins. Here, the expression and function of the testis-enriched proteins tPlus3a and tPlus3b have been examined. Both proteins share the conserved Plus3 domain of Rtf1. RNAseq analysis using RNA from mutant testes revealed that tPlus3a and tPlus3b likely contribute to the regulation of transcription in spermatocytes. Furthermore, genes which also depend on tBRD-1 function were identified. We hypothesise that tPlus3a and tPlus3b regulate a group of genes overlapping with tBRD-1-depending genes but not with genes depending on tTAFs. tPlus3a and tPlus3b might therefore contribute to diversification of transcriptional regulation in spermatocytes

Identification of male- and female-specific olfaction genes in antennae of the oriental fruit fly (Bactrocera dorsalis)

The oriental fruit fly (Bactrocera dorsalis) is a species of tephritid fruit fly, endemic to Southeast Asia but also introduced to many regions of the US, and it is one of the major pest species with a broad host range of cultivated and wild fruits. Although males of B. dorsalis respond strongly to methyl eugenol and this is used for monitoring and estimating populations, the molecular mechanism of the oriental fruit fly olfaction has not been elucidated yet. Therefore, in this project, using next generation sequencing technologies, we sequenced the transcriptome of the antennae of male and female adults of B. dorsalis. We identified a total of 20 candidate odorant binding proteins (OBPs), 5 candidate chemosensory proteins (CSPs), 35 candidate odorant receptors (ORs), 12 candidate ionotropic receptors (IRs) and 4 candidate sensory neuron membrane proteins (SNMPs). The sex-specific expression of these genes was determined and a subset of 9 OR genes was further characterized by qPCR with male and female antenna, head, thorax, abdomen, leg and wing samples. In the male antennae, 595 genes showed a higher expression, while 128 genes demonstrated a higher expression in the female antennae. Interestingly, 2 ORs (BdorOR13 and BdorOR14) were highly and specifically expressed in the antennae of males, and 4 ORs (BdorOR13, BdorOR16, BdorOR18 and BdorOR35) clustered with DmOR677, suggesting pheromone reception. We believe this study with these antennae-enriched OBPs, CSPs, ORs, IRs and SNMPs can play an important role in the detection of pheromones and general odorants, and so in turn our data improve our current understanding of insect olfaction at the molecular level and provide important information for disrupting the behavior of the oriental fruit fly using chemical communication methods

Lack of phenotypic and evolutionary cross-resistance against parasitoids and pathogens in Drosophila melanogaster

BackgroundWhen organisms are attacked by multiple natural enemies, the evolution of a resistance mechanism to one natural enemy will be influenced by the degree of cross-resistance to another natural enemy. Cross-resistance can be positive, when a resistance mechanism against one natural enemy also offers resistance to another; or negative, in the form of a trade-off, when an increase in resistance against one natural enemy results in a decrease in resistance against another. Using Drosophila melanogaster, an important model system for the evolution of invertebrate immunity, we test for the existence of cross-resistance against parasites and pathogens, at both a phenotypic and evolutionary level.MethodsWe used a field strain of D. melanogaster to test whether surviving parasitism by the parasitoid Asobara tabida has an effect on the resistance against Beauveria bassiana, an entomopathogenic fungus; and whether infection with the microsporidian Tubulinosema kingi has an effect on the resistance against A. tabida. We used lines selected for increased resistance to A. tabida to test whether increased parasitoid resistance has an effect on resistance against B. bassiana and T. kingi. We used lines selected for increased tolerance against B. bassiana to test whether increased fungal resistance has an effect on resistance against A. tabida.Results/ConclusionsWe found no positive cross-resistance or trade-offs in the resistance to parasites and pathogens. This is an important finding, given the use of D. melanogaster as a model system for the evolution of invertebrate immunity. The lack of any cross-resistance to parasites and pathogens, at both the phenotypic and the evolutionary level, suggests that evolution of resistance against one class of natural enemies is largely independent of evolution of resistance against the other

Female remating in Drosophila ananassae: bidirectional selection for remating speed

In Drosophila ananassae, artificial selection was carried out for fast and slow remating speed for 10 generations. Response to selection resulted in rapid divergence in remating time in each of two replicates of both fast and slow lines. There were significant differences in mean remat-ing time in females among fast, slow, and control lines. Regression coefficients for both fast and slow lines are significantly different from zero. The realized heritability over 10 genera-tions of selection is from 0.26 to 0.33 for two replicates of fast line and from 0.23 to 0.27 for two replicates of slow line. These findings suggest that female remating time in D. ananassae is under polygenic control. Remating frequency of females showed a correlated response in both fast and slow lines. At generation 10, correlated response to selection was also investigated. Mating propensity of D. ananassae of fast and slow lines was observed in an Elens-Wattiaux mating chamber. Fifteen pairs per test showed that on the average, the fast lines (11.20, 11.60) were more successful in mating than those of slow (6.40, 5.60) and control (8.00) lines. Pro-ductivity of once-mated females was measured in terms of number of progeny produced per fe-male and the results of productivity analysis indicate that females of fast lines (157.83, 130.83) produced more progeny compared with slow (72.70, 85.83) and control (109.23) lines

Adventures in Time and Space: What Shapes Behavioural Decisions in Drosophila melanogaster?

Variation in behaviour can be observed both between individuals, based on their condition and experience as well as between populations due to sources of heterogeneity in the environment. These behavioural differences have evolved as a result of natural and sexual selection where different strategies may be favoured depending on the costs and benefits associated with those behaviours. In this thesis I examine two sources of heterogeneity within the environment and their behavioural consequences: how spatial complexity mediates sexual selection over time, and how inter and intraspecific signals and individual condition influence social oviposition behaviour. By increasing spatial complexity, we were able to manipulate male-female interaction rate which in turn influenced courtship behaviour and male-induced harm, the consequence of this was an increase in female fecundity especially in the later days of the assay and no change in offspring fitness. These results supported the idea that spatial complexity is able to mediate sexual selection through decreased harm to females. Oviposition decisions are of high consequence to an individual’s fitness and can be shaped by many environmental conditions. Instead of expending energy to evaluate all their different costs and benefits of the conditions of potential oviposition sites females can chose to rely on the signals left by others, in this case it would be beneficial for females to identify signals most like themselves. While we found females oviposited with individuals of the same species and diet, when given the option they showed more interest in and laid more eggs on media that previously held virgin males, bringing into question many assumptions of copying behaviour. In Drosophila melanogaster the only control females have over their offspring is who they mate with and where they oviposit their eggs, thus, these two factors can have a long-lasting impact on individual fitness for future generations. It is also important to consider how the standard lab environment may be shaping these behaviours, and the consequences this has for the evolutionary trajectory of lab populations

The onset of homologous chromosome pairing during Drosophila melanogaster embryogenesis.

We have determined the position within the nucleus of homologous sites of the histone gene cluster in Drosophila melanogaster using in situ hybridization and high-resolution, three-dimensional wide field fluorescence microscopy. A 4.8-kb biotinylated probe for the histone gene repeat, located approximately midway along the short arm of chromosome 2, was hybridized to whole-mount embryos in late syncytial and early cellular blastoderm stages. Our results show that the two homologous histone loci are distinct and separate through all stages of the cell cycle up to nuclear cycle 13. By dramatic contrast, the two homologous clusters were found to colocalize with high frequency during interphase of cycle 14. Concomitant with homolog pairing at cycle 14, both histone loci were also found to move from their position near the midline of the nucleus toward the apical side. This result suggests that coincident with the initiation of zygotic transcription, there is dramatic chromosome and nuclear reorganization between nuclear cycles 13 and 14