Cryptic reproductive isolation in the Drosophila simulans species complex

Forms of reproductive isolation that act after copulation but before fertilization are potentially important components of speciation, but are studied only infrequently. We examined postmating, prezygotic reproductive isolation in three hybridizations within the Drosophila simulans species complex. We allowed females to mate only once, observed and timed all copulations, dissected a subset of the females to track the storage and retention of sperm, examined the number and hatchability of eggs laid after insemination, counted all progeny produced, and measured the longevity of mated females. Each of the three hybridizations is characterized by a different set of cryptic barriers to heterospecific fertilization. When D. simulans females mate with D. sechellia males, few heterospecific sperm are transferred, even during long copulations. In contrast, copulations of D. simulans females with D. mauritiana males are often too short to allow sperm transfer. Those that are long enough to allow insemination, however, involve the transfer of many sperm, but only a fraction of these heterospecific sperm are stored by females, who also lay fewer eggs than do D. simulans females mated with conspecific males. Finally, when D. mauritiana females mate with D. simulans males, sperm are transferred and stored in abundance, but are lost rapidly from the reproductive tract and are therefore used inefficiently. These results add considerably to the list of reproductive isolating mechanisms in this well-studied clade and possibly to the list of evolutionary processes that could contribute to their reproductive isolation.     

Sexual isolation between Drosophila melanogaster females and D. simulans males: sexual maturation of males

Isofemale lines of D. simulans were examined to determine the age of sexual maturity of males with conspecific females, and for the frequency of hybridization with D. melanogaster females. Males started to mature sexually on the first day after eclosion but their ability to mate slowly increased during the following day. The estimates of both the age sexual maturation started and the switch from immature to mature males were strongly dependent on the female genotypes used in the tests. No clear differences in speed of maturation were apparent between male lines. In contrast, differences in frequency of hybridization with D. melanogaster females did occur. From the above results it is concluded that the differential hybridization success of male D. simulans lines is not related to the speed at which males mature sexually.

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Résumé Des lignées isofemelles de D. simulans ont été examinées pour déterminer l'âge de la maturité sexuelle des mâles avec des femelles conspécifiques et pour établir la fréquence de l'hybridation avec des femelles de D. melanogaster. Les mâles ont commencé à être sexuellement mûrs le premier jour après l'émergence, mais leur aptitude à la copulation a augmenté lentement pendant le jour suivant. Les estimations, tant de l'âge du début de la maturation sexuelle que de l'âge du passage de mâle immature à mâle sexuellement mûr dépendaient étroitement des génotypes des femelles utilisées dans les expériences. Il n'y avait pas de différences nettes entre les lignées de mâles. Par contre, des différences dans les fréquences d'hybridation avec les femelles de D. melanogaster ont été observées. De ces résultats, on peut conclure que les différences dans la réussite des hybridations des lignées de mâles de D. simulans n'étaient pas dues à la vitesse de maturation sexuelle des mâles.

     

Quantitative trait loci for sexual isolation between Drosophila simulans and D. mauritiana

Sexual isolating mechanisms that act before fertilization are often considered the most important genetic barriers leading to speciation in animals. While recent progress has been made toward understanding the genetic basis of the postzygotic isolating mechanisms of hybrid sterility and inviability, little is known about the genetic basis of prezygotic sexual isolation. Here, we map quantitative trait loci (QTL) contributing to prezygotic reproductive isolation between the sibling species Drosophila simulans and D. mauritiana. We mapped at least seven QTL affecting discrimination of D. mauritiana females against D. simulans males, three QTL affecting D. simulans male traits against which D. mauritiana females discriminate, and six QTL affecting D. mauritiana male traits against which D. simulans females discriminate. QTL affecting sexual isolation act additively, are largely different in males and females, and are not disproportionately concentrated on the X chromosome: The QTL of greatest effect are located on chromosome 3. Unlike the genetic components of postzygotic isolation, the loci for prezygotic isolation do not interact epistatically. The observation of a few QTL with moderate to large effects will facilitate positional cloning of genes underlying sexual isolation.     

Interspecific sexual isolation and phylogeny among different members of the Drosophila bipectinata species complex

The degree and pattern of sexual isolation among closely related species can be utilized for predicting the direction of evolution and deriving a phylogeny. The Drosophila bipectinata species complex is a group of four morphologically very similar species, belonging to the ananassae subgroup of the melanogaster species group. Sexual isolation among the members of this complex was studied by employing the male-choice technique. To test the difference between the homogamic and heterogamic matings, chi-square values were calculated under the assumption of random mating and the degree of sexual isolation was measured by estimating isolation index. To test the difference in proportions of heterogamic matings between reciprocal crosses, normal deviate (z) was calculated. The direction of evolution among these species has been discussed in the light of models proposed by different workers. Our results suggest that while D. bipectinata, Drosophila parabipectinata and Drosophila malerkotliana are closely related to each other, D. pseudoananassae is distantly related to these three species. The species pairs D. bipectinata and D. parabipectinata and D. parabipectinata and D. malerkotliana show asymmetrical sexual isolation. Based on this, it is suggested that D. bipectinata and D. malerkotliana share a common ancestor where as D. parabipectinata has been derived from D. bipectinata. Their phylogenetic relationship has been discussed in the light of phylogenies suggested by earlier workers.     

Intraspecific sexual isolation in Drosophila.

Intraspecific sexual isolation was examined among wild strains of Drosophila malerkotliana, D. parabipectinata and D. pseudoananassae by multiple-choice method in Elens-Wattiaux mating chamber. In D. pseudoananassae, mating between two strains tested was random and isolation estimate was close to one. In one out of 6 crosses, involving geographic strains of D. malerkotliana, there was significant deviation from randomness and isolation estimate remained low which shows non-random (preferential or positive assortative) mating. In D. parabipectinata, the deviation from randomness was statistically significant due to higher number of homogamic matings in three crosses involving wild strains derived from geographically distant places and isolation estimate remained low in these crosses. The results provide evidence for incipient sexual isolation within D. malerkotliana and D. parabipectinata as a result of genetic divergence.     

Genetics of sexual isolation based on courtship song between two sympatric species: Drosophila ananassae and D. pallidosa

Sexual isolation has been considered one of the primary causes of speciation and its genetic study has the potential to reveal the genetics of speciation. In Drosophila, the importance of courtship songs in sexual isolation between closely related species has been well investigated, but studies analysing the genetic basis of the difference in the courtship songs associated with sexual isolation are less well documented. Drosophila ananassae and Drosophila pallidosa are useful for studies of sexual isolation, because of their sympatric distribution and absence of postmating isolation. Courtship songs are known to play a crucial role in sexual isolation between these two species, and the female discrimination behaviour against the courting male has been revealed to be controlled by a very narrow region on the second chromosome. In this study we investigated the genetic basis controlling the song differences associated with their sexual isolation, using intact and wingless males with chromosomes substituted between species. The results obtained from F₁ hybrid males between these species indicate the dominance of the song characters favoured by D. pallidosa females. In addition, the results obtained from backcross F₂ males indicate that chromosome 2 had a major effect on the control of the song characters associated with sexual isolation.     

Genetics and the environment in interspecific competition: a study using the sibling species Drosophila melanogaster and Drosophila simulans

The outcome of interspecific competition of two closely related species may depend upon genetic variation in the two species and the environment in which the experiment is carried out. Interspecific competition in the two sibling species, Drosophila melanogaster and D. simulans, is usually investigated using longterm laboratory stocks that often have mutant markers that distinguish them. To examine competition in flies that genetically more closely resemble flies in nature, we utilized freshly caught wildtype isofemale lines of the two species collected at the same site in San Carlos, Mexico. Under ordinary laboratory conditions, D. melanogaster always won in competition. However, in hotter and drier conditions, D. simulans competed much more effectively. In these environmental conditions, there were genetic differences in competitive ability among lines with the outcome of competition primarily dependent upon the line of D. melanogaster used but in some cases also influenced by the line of D. simulans used. Differences in the measures of productivity and developmental time did not explain the differences in competitive ability among lines. This suggests that the outcome of competition was not due to differences in major fitness components among the isofemale lines but to some other attribute(s) that influenced competitive ability. When lines of flies were combined, the outcome of competition was generally consistent with competitive outcomes between pairs of lines. In several cases, the combination of lines performed better than the best of the constituent lines, suggesting that competitive ability was combined heterotically and that the total amount of genetic variation was important in the outcome of interspecific competition.     

The population genetics of the origin and divergence of the Drosophila simulans complex species

The origins and divergence of Drosophila simulans and close relatives D. mauritiana and D. sechellia were examined using the patterns of DNA sequence variation found within and between species at 14 different genes. D. sechellia consistently revealed low levels of polymorphism, and genes from D. sechellia have accumulated mutations at a rate that is approximately 50% higher than the same genes from D. simulans. At synonymous sites, D. sechellia has experienced a significant excess of unpreferred codon substitutions. Together these observations suggest that D. sechellia has had a reduced effective population size for some time, and that it is accumulating slightly deleterious mutations as a result. D. simulans and D. mauritiana are both highly polymorphic and the two species share many polymorphisms, probably since the time of common ancestry. A simple isolation speciation model, with zero gene flow following incipient species separation, was fitted to both the simulans/mauritiana divergence and the simulans/sechellia divergence. In both cases the model fit the data quite well, and the analyses revealed little evidence of gene flow between the species. The exception is one gene copy at one locus in D. sechellia, which closely resembled other D. simulans sequences. The overall picture is of two allopatric speciation events that occurred quite near one another in time.     

Quantitative trait loci for cuticular hydrocarbons associated with sexual isolation between Drosophila simulans and D. sechellia

The identification of genes with large effects on sexual isolation and speciation is an important link between classic evolutionary genetics and molecular biology. Few genes that affect sexual isolation and speciation have been identified, perhaps because many traits influencing sexual isolation are complex behaviors. Cuticular hydrocarbons (CHs) of species of the Drosophila melanogaster group play a large role in sexual isolation by functioning as contact pheromones influencing mate recognition. Some of the genes that play key roles in determining species-specific CHs have been identified. We have performed separate quantitative trait locus (QTL) analyses of 7-tricosene (7-T) and 7,11-heptacosadiene (7,11-HD), the two major female CHs differing between D. simulans and D. sechellia. We find that approximately 40% of the phenotypic variance in each CH is associated with two to four chromosomal regions. A region on the right arm of chromosome 3 contains QTL that affect both traits, but other QTL are in distinct chromosomal regions. Epistatic interactions were detected between two pairs of QTL for 7,11-HD such that if either were homozygous for the D. simulans allele, the fly was similar to D. simulans in phenotype, with a low level of 7,11-HD. We discuss the location of these regions with regard to candidate genes for CH production, including those for desaturases.     

Genetics of Drosophila simulans male mating discrimination in crosses with D. melanogaster

The genetic bases of sexual isolation between Drosophila melanogaster and D. simulans have been mainly studied in females, and there is little information about the role of the males in interspecific mating discrimination. Using D. simulans synthetic lines with compound chromosomes from a population of the Seychelles Islands (high frequency of interspecific mating) and a multimarker strain (low frequency), we show that D. simulans males play an important role in discriminating D. melanogaster females. The genetics of male discrimination fits well with the inheritance mode of a single locus, dominant for sexual isolation, located in chromosome II near the net mutation (2L-0.0). The heterospecific mating success of the male was not related to his sexual vigor. The specific load of male cuticular hydrocarbons was counted as a possible source of discrimination used by the D. melanogaster female.     

Genetics of sexual behaviour in Drosophila

The analysis of genetics of behaviour within and between species provides important clues about the forces shaping the evolution of behavioural genes. In Drosophila, a number of key processes such as emergence from the pupal case, locomotor activity, feeding, olfaction and aspects of mating behaviour are under circadian regulation. Genes controlling sexual behaviour are likely to control species specific differences in courtship that are involved in reproductive isolation of closely related species. Courtship in Drosophila is characterized by a series of stereotyped behaviours that lead to copulation and more than 30 genes have been identified through mutations that affect one or more of these elements. Although curiosity about behavioural differences between the sexes undoubtedly predates recorded history, little efforts have been made to uncover the molecular basis of male and female courtship. The brain and nervous system functions that underlie sex-specific behaviour are of obvious importance to all animals including humans. To understand behaviour related to sex it is important to distinguish those aspects that are controlled genetically. The isolation and analysis of Drosophila mutants with altered sexual orientation lead to the identification of novel branches in the sex-determination cascade, which govern the sexually dimorphic development of the nervous system.     

A quantitative genetic analysis of male sexual traits distinguishing the sibling species Drosophila simulans and D. sechellia

A quantitative trait locus (QTL) genetic analysis of morphological and reproductive traits distinguishing the sibling species Drosophila simulans and D. sechellia was carried out in a backcross design, using 38 markers with an average spacing of 8.4 cM. The direction of QTL effects for the size of the posterior lobe was consistent across the identified QTL, indicating directional selection for this trait. Directional selection also appears to have acted on testis length, indicating that sexual selection may have influenced many reproductive traits, although other forms of directional selection cannot be ruled out. Sex comb tooth number exhibited high levels of variation both within and among isofemale lines and showed no evidence for directional selection and, therefore, may not have been involved in the early speciation process. A database search for genes associated with significant QTL revealed a set of candidate loci for posterior lobe shape and size, sex comb tooth number, testis length, tibia length, and hybrid male fertility. In particular, decapentaplegic (dpp), a gene known to influence the genital arch, was found to be associated with the largest LOD peak for posterior lobe shape and size.     

Bacteria-induced sexual isolation in Drosophila

Commensal bacteria can induce sexual isolation between populations of Drosophila. This phenomenon has implications for speciation, and raises questions about its behavioral and developmental mechanisms, which are not yet known. In this Extra View, we discuss related work by others, bearing directly on these issues, and we speculate about how bacteria might influence fly behavior.

There are many reports of interaction between Drosophila and their microbiota that significantly impacts mating preferences. Sexual isolation can be enhanced or reduced by altering the culture media, or the microbiota inhabiting those media. More dramatically, the endoparasite Wolbachia has induced strong mate preferences in some instances. While a sudden, ecologically induced shift in mating preferences falls far short of the changes required for speciation, it might be a first step in that direction.

We hypothesize that bacteria-induced sexual isolation is caused by chemosensory cues. In our experiments, bacteria altered the profile of cuticular hydrocarbons, which function as sex pheromones. Commensal bacteria may act directly on these hydrocarbons, or they may affect their synthesis. Alternatively, bacterial metabolites might perfume the flies in ways that affect mate choice. In that event, habituation or conditioning likely plays a role.     

Sexual isolation between Drosophila simulans and D. mauritiana: D. simulans females do not discriminate against intact-wing D. mauritiana males

Journal of Ethology - Female mate choice plays an important role in sexual isolation. The present study examined sexual isolation using D. simulans and D. mauritiana in conditions...     

The evolution of reproductive isolation in the Drosophila yakuba complex of species

In the Drosophila melanogaster subgroup, the yakuba species complex, D. yakuba, D. santomea and D. teissieri have identical mitochondrial genomes in spite of nuclear differentiation. The first two species can be readily hybridized in the laboratory and produce fertile females and sterile males. They also form hybrids in natural conditions. Nonetheless, the third species, D. teissieri, was thought to be unable to produce hybrids with either D. yakuba or D. santomea. This in turn posed the conundrum of why the three species shared a single mitochondrial genome. In this report, we show that D. teissieri can indeed hybridize with both D. yakuba and D. santomea. The resulting female hybrids from both crosses are fertile, whereas the hybrid males are sterile. We also characterize six isolating mechanisms that might be involved in keeping the three species apart. Our results open the possibility of studying the history of introgression in the yakuba species complex and dissecting the genetic basis of interspecific differences between these three species by genetic mapping.     

Male pheromone polymorphism and reproductive isolation in populations of Drosophila simulans

The dominant cuticular hydrocarbons (HC) in Drosophila simulans are 7‐tricosene (7‐T) and 7‐pentacosene (7‐P). The 7‐T is the major HC in East Africa and in other continents. In West Africa, D. simulans is very rare and displays 7‐P as the major compound. We studied three D. simulans strains from Egypt (Eg), Sao‐Tome (ST), and Cameroon (Cam), with 7‐T, intermediary or 7‐P phenotypes. HC profiles of ST and Cam female differed slightly from corresponding male profiles; females had more 7‐T and less 7‐P. Varying temperature affected all HCs (even those with 27 and 29 carbons)‐not just 7‐T and 7‐P; there was no clear relationship between HC phenotype and resistance to desiccation. We report reproductive isolation between Eg and ST and Eg and Cam (but not between ST and Cam), which is due to Eg and Cam female preferences for their own males. In conclusion, our findings do support divergence of D. simulans populations from West Africa for both pheromonal profile and mating preference.