No evidence for minority male mating advantage in wild type strains of Drosophila ananassae tested in multiple-choice experiments

Minority male mating advantage was tested in wild type strains of Drosophila ananassae through multiple-choice experiments. Mating success of two types of flies present in five different ratios was scored by direct observation in an Elens-Wattiaux mating chamber. We found no evidence for minority male mating advantage in wild type strains of D. ananassae. The relative mating success of two types of females was also compared in the multiple-choice experiments at different ratios; there was no evidence for a rare female effect. Further, there was similarity in the results of experiments employing different methods. The total number of homogamic and heterogamic matings was obtained by combining the data (all five ratios) from each experiment. Homogamic matings were significantly more frequent than heterogamic ones, which demonstrates preferential mating between males and females of the same strain; this was also supported by a lower isolation estimate. There was also a significant difference in the degree of mating preference between the two strains.     

Evolution of mating isolation between populations of Drosophila ananassae

Prezygotic mating isolation has been a major interest of evolutionary biologists during the past several decades because it is likely to represent one of the first stages in the transition from populations to species. Mate discrimination is one of the most commonly measured forms of prezygotic isolation and appears to be relatively common among closely related species. In some cases, it has been used as a measure to distinguish populations from subspecies, races, and sister species, yet the influences of various evolutionary mechanisms that may generate mate discrimination are largely unknown. In this study, we measured the level and pattern of mate discrimination among 18 populations of a cosmopolitan drosophilid species, Drosophila ananassae , from throughout its geographical range and its sister species, Drosophila pallidosa, which has a restricted geographical distribution in the South Pacific Islands. In addition, we measured genetic differentiation between all 18 populations using mitochondrial DNA polymorphism data. Mate discrimination varies considerably throughout the species range, being higher among populations outside the ancestral Indonesian range, and highest in the South Pacific. Our results suggest that colonization and genetic differentiation may have an influence on the evolutionary origin of mate discrimination. Our phylogeographical approach clarifies the ancestral relationships of several populations from the South Pacific that show particularly strong mate discrimination and suggests that they may be in the early stages of speciation. Furthermore, both the genetic and behavioral results cast doubt on the status of D. pallidosa as a good species.     

Evidence for minority male mating success and minority female mating disadvantage in Drosophila ananassae

Frequency-dependent mating success was tested for three pairs of wild-type and mutant strains of Drosophila ananassae, MY and yellow body color (y), PN and claret eye color (ca), and TIR and cut wing (ct). The two strains of each pair were chosen for their approximately equal mating propensities. Multiple-choice experiments, using different experimental procedures, were employed. The tests were carried out by direct observation in Elens-Wattiaux mating chambers with five different sex ratios (4:16, 8:12, 10:10, 12:8, and 16:4). There was no assortative mating and sexual isolation between the strains, based on 2 x 2 contingency chi2 analysis and isolation estimate values. One-sided rare male mating advantages were found in two experiments, one for ca males and the other for wild-type males (TIR). However, no advantage was found for rare males in the experiment with MY and y flies. Mating disadvantages for rare females were found for sex-linked mutants (y and ct). Two different observational methods (removal or direct observation of mating pairs) imparted no overall significant effects on the outcome of the frequency-dependent mating tests.     

Variation in mating propensity and fertility in isofemale strains of Drosophila ananassae

Mating propensity and fertility were studied in five laboratory strains of Drosophila ananassae which were established from single females collected from different geographical localities. The results show statistically significant variation among different isofemale lines with respect to mating propensity and fertility. The strains showing greater sexual activity produce more progeny. Thus, there is a positive correlation between mating activity and fertility in D. ananassae. The comparison of mating frequencies of strains and their hybrids reveals the existence of heterosis and reciprocal effects. The data suggest that the males are more subject to intrasexual selection.     

Interracial hybridization in Drosophila ananassae

Crosses were made between strains of Drosophila ananassae, homozygous for the ST or AL gene sequences in the second chromosome, and originating from geographically distant localities in India. All interracial crosses were maintained separately in culture bottles for ten generations. After ten generations it was observed that the inversion heterozygotes were heterotic. Thus it was found that interracial hybridization does not lead to breakdown of heterosis. Therefore, it has been suggested that localised coadaptation is absent in the natural populations of Indian D. ananassae. Thus D. ananassae does not show the pattern of some other species.     

Behavioral genetics of Drosophila ananassae

Drosophila ananassae has a unique status among Drosophila species because of certain peculiarities in its genetic behavior. The most unusual feature of this species is its relatively high frequency of spontaneous male recombination. The results of studies on non-sexual behavior, such as phototactic responses, eclosion rhythm, and preferences for oviposition and pupation sites, lead us to suggest that this behavior is under polygenic control, with a substantial amount of additive genetic variation. Sexual isolation has been reported in D. ananassae with the degree of such isolation being stronger in isofemale lines than in natural populations. The significant variations seen in the mating propensity of several isofemale strains, inversion karyotypes and wild type strains, the diminishing effects of certain mutations on the sexual activity of males, and the positive responses to selection for high and low mating propensity point to a genetic control of sexual behavior in D. ananassae. Males contribute more to variation and thus are more subject to intrasexual selection than females. There is a positive correlation between sternopleural bristle number, mating propensity and fertility in D. ananassae. This correlation between morphometric traits and mating success suggests that larger flies are more successful in mating than smaller ones. There is also evidence for adaptive plasticity and a trade-off between longevity and productivity in D. ananassae. Rare, specific courtship song parameters that provide males with a mating advantage have also been reported in different geographic strains of D. ananassae. The remating behavior of males and females, sperm displacement, and the bi-directional selection for female remating speed indicate that post-mating behavior in this species may also be under genetic control. The occurrence of size assortative mating further indicates that there is size-dependent sexual selection in D. ananassae.     

Genetic analysis of female mating recognition between Drosophila ananassae and Drosophila pallidosa: application of interspecific mosaic genome lines

Drosophila ananassae and Drosophila pallidosa are closely related species that can produce viable and fertile hybrids of both sexes, although strong sexual isolation exists between the two species. Females are thought to discriminate conspecific from heterospecific males based on their courtship songs. The genetic basis of female discrimination behavior was analyzed using isogenic females from interspecific mosaic genome lines that carry homozygous recombinant chromosomes. Multiple regression analysis indicated a highly significant effect of the left arm of chromosome 2 (2L) on the willingness of females to mate with D. ananassae males. Not only 2L but also the left arm of chromosome X (XL) and the right arm of chromosome 3 (3R) had significant effects on the females' willingness to mate with D. pallidosa males. All regions with strong effects on mate choice have chromosome arrangements characterized by species-specific inversions. Heterospecific combinations of 2L and 3R have previously been suggested to cause postzygotic reproductive isolation. Thus, genes involved in premating as well as postmating isolation are located in or near chromosomal inversions. This conclusion is consistent with the recently proposed hypothesis that "speciation genes" accumulate at a higher rate in non-recombining genome regions when species divergence occurs in the presence of gene flow.     

Recombination between heterozygous inversions in Drosophila ananassae

The rate of recombination between linked inversions of the third chromosome of Drosophila ananassae has been investigated by means of the salivary gland smear technique. Different homozygous strains of the species were utilised during the present investigation. The data show that the crossover rate between heterozygous inversions of the third chromosome of D. ananassae varies among different strains. This suggests that recombination between heterozygous inversions is influenced by genic (strain) factors.     

Size Dependent Sexual Selection in Drosophila ananassae

Mate choice based on body size is widespread and can have numerous consequences. We present data, which show the effect of male and female body size on sexual selection in Drosophila ananassae. The relationships between wing size, locomotor activity, mating latency, courtship pattern, fertility and mating success were studied. Mating latency was negatively correlated with wing length and with locomotor activity, while wing length and locomotor activity was positively correlated in males as well as in females. In female- and male-choice, we found that mate choice influenced size-assortative mating by: (1) large and small males preferring to mate with large females, (2) large males successfully competing for large females, leaving small males to mate with small females. Males increased their reproductive success by mating with large and more fecund females. In addition, in pairs of long/short winged flies, long winged flies courted and mated more successfully than short winged flies and they also have longer duration of copulation and more progeny than short winged flies. We found sterile mating in pairs of small winged males and females.     

Genetic differentiation in natural populations of Drosophila ananassae

Inversion polymorphism has been studied in three natural populations of Drosophila ananassae. The three cosmopolitan inversions have been detected in all the three populations analysed. Some quantitative variations in the inversion frequencies seem to exist and the level of inversion heterozygosity varies between the different populations.     

Search for species-specific mating signal in courtship songs of sympatric sibling species,Drosophila ananassae and D. pallidosa

Sexual isolation is one of the most important mechanisms that may lead to speciation. Drosophila ananassae and D. pallidosa are useful for the study of sexual isolation, because of their sympatric distribution and no postmating isolation between them. Courtship songs are considered to play a crucial role in sexual isolation between D. ananassae and D. pallidosa. We recorded and analyzed male courtship songs of D. ananassae and D. pallidosa for eight and four geographical strains, respectively. Courtship behaviors of the two species were consistent with those previously described, however, male's middle leg shaking, which had not before been described, was observed in both species. Males sing by wing vibration only during courtship. Their song oscillograms were distinct between species, but those of conspecific strains were very similar, in spite of their different geographical derivation. We found species-specificity in burst length, pulse length, cycle number in a pulse, and frequency spectra of bursts; these results suggest that these song parameters may play a role in mate recognition that enforces their sexual isolation. The specific values of interpulse interval, cycle number in a pulse and intrapulse frequency were involved with the determination of specificity in frequency spectra of bursts. We discussed the possibility that the specific frequency spectra of bursts are recognized by females as the species-specific signal rather than each parameter individually.     

On intra- and interchromosomal associations in Drosophila ananassae

In Drosophila ananassae, three cosmopolitan inversions are very common in natural populations. Chromosomal polymorphism due to these inversions often persists when strains are maintained in the laboratory. A chromosomal analysis of a number of strains was made. Data on the frequencies of different gene sequences will be described separately. During the present investigation the same data have been analyzed in order to test the intra- and interchromosomal interactions in D. ananassae. The results reveal that interchromosomal interactions do not occur with respect to viability as unlinked inversions are associated randomly. However, the linked inversions of the third chromosome show non-random associations maintained by epistatic gene interaction and suppression of crossing over.     

Population and behaviour genetics of Drosophila ananassae

Drosophila ananassae is a cosmopolitan and domestic species. It occupies a nuique status among the Drosophila species due to certain peculiarities in its genetic behaviour. The most unusual feature of this species is spontaneous male recombination in appreciable frequency. The present review summarises the work done on population and behaviour genetics of D. ananassae from India. Population dynamics of three cosmopolitan inversions has been studied in Indian population of D. ananassae and it is evident from the results that there is a considerable degree of genetic divergence at the level of inversion polymorphism. In general, the populations from south India show more differentiation than those from the north. These three cosmopolitan inversions, which are coextensive with the species, exhibit heterosis. Interracial hybridization does not lead to beaakdown of heterosis, which suggests that evidence for coadaptation is lacking in geographic populations of D. ananassae. Heterosis appears to be simple luxuriance rather than populational heterosis (coadaptation). Unlinked inversions occur in random associations, indicating no interchromosomal interactions. However, two inversions of the third chromosome often show strong linkage disequilibrium in laboratory populations, which is due to epistatic gene interaction and suppression of crossing-over. Genetic variations for certain allozyme polymorphism and sternoleural bristle phenotypes in Indian populations of D. ananassae have also been observed.A number of investigations have also been carried out on certain aspects of behaviour genetics of Indian D. ananassae. There is evidence for sexual isolation within D. ananassae. Significant variations in mating propensity of several isofemale strains, inversion karyotypes, the diminishing effects of certain mutations on sexual activity of males and positive response to selection for high and low mating propensity provide evidence for genetic control of sexual behaviour in D. ananassae. Males contribute more to variation and thus are more subject to intra-sexual selection than females. Evidence for rare male mating advantage has also been presented. Geographic strains of D. ananassae show variation with respect to oviposition site preference. The results of studies on pupation site preference, which is an important component of larval behaviour, suggest that larval pupation behaviour in D. ananassae is under polygenic control with a substantial amount of additive genetic variation.     

A site specific increase in recombination in Drosophila ananassae

The e65 pi; bri ru stock of Drosophila ananassae produced an extremely high rate of recombination in males when made heterozygous with any one of the wild type stocks. We analyzed and characterized the genetic factors which caused this phenomenon. We show that the second chromosome of the e65 pi; bri ru stock carries an enhancer of male recombination. The enhancer, En(2)-ep, is located between Om(2C) and Arc. The enhancement of meiotic recombination both in males and females was also observed at the specific region between Om(2C) and Arc on 2L. The magnitude of increased recombination was 30-40 fold in males and 13-30 fold in females. The relation between the hotspot of recombination in both sexes and the enhancer of male recombination is discussed.     

Disruptive selection for sternopleural bristle phenotypes in Drosophila ananassae

Disruptive selection is potentially critical in maintaining variation and initiating speciation and plays an important role in the organization of genetic variability in natural populations. It occurs when extreme phenotypes have a fitness advantage over intermediate phenotypes. Disruptive selection for high and low numbers of sternopleural bristles in Drosophila ananassae was applied for 12 generations to test its effect in induction of behavioural isolation. Pattern of mating between flies of high and low lines was tested in an Elens-Wattiaux mating chamber by using a multiple-choice technique after G(5) and G(12). Data was analyzed by calculating χ(2) under the assumption of random mating to test the difference between homoand heterogamic matings, and sexual isolation was tested by calculating the isolation estimate. The results show that there is no evidence for sexual isolation in G(5) and G(12). The realized heritability, standard error of regression coefficient, and t values suggest that disruptive selection for sternopleural bristle phenotypes was effective, but it does not lead to behavioral isolation in D. ananassae. Rather, it induces differences in mating propensity, which is influenced by sternopleural bristle phenotypes.     

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.     

Experimental Evidence for Nutrition Regulated Stress Resistance in Drosophila ananassae

Background

The amount and quality of nutrients consumed by organisms have a strong impact on stress resistance, life-history traits and reproduction. The balance between energy acquisition and expenditure is crucial to the survival and reproductive success of animals. The ability of organisms to adjust their development, physiology or behavior in response to environmental conditions, called phenotypic plasticity, is a defining property of life. One of the most familiar and important examples of phenotypic plasticity is the response of stress tolerance and reproduction to changes in developmental nutrition. Larval nutrition may affect a range of different life-history traits as well as responses to environmental stress in adult.

Principal Findings

Here we investigate the effect of larval nutrition on desiccation, starvation, chill-coma recovery, heat resistance as well as egg to adult viability, egg production and ovariole number in Drosophila ananassae. We raised larvae on either protein rich diet or carbohydrate rich diet. We found that flies consuming protein rich diet have higher desiccation and heat shock resistance whereas flies developed on carbohydrate rich diet have higher starvation and cold resistance. Egg production was higher in females developed on protein rich diet and we also found trade-off between egg production and Egg to adult viability of the flies. Viability was higher in carbohydrate rich diet. However, sex specific viability was found in different nutritional regimes. Higher Egg production might be due to higher ovariole number in females of protein rich diet.

Conclusion

Thus, Drosophila ananassae adapts different stress tolerance and life-history strategies according to the quality of the available diet, which are correlated with phenotypic adjustment at anatomical and physiological levels.