Sex Comb Tooth Number Research Articles

Long‐term coexistence of a hybridization‐derived population of Drosophila parapallidosa with closely related Drosophila ananassae (Diptera: Drosophilidae)

AbstractA 2012–13 survey on Penang Island, Malaysia, revealed the existence of both Drosophila ananassae and Drosophila parapallidosa, the latter of which carries chromosomes Y and 4 from D. ananassae and thus is of hybrid origin. We collected the flies again from the same location in 2018. The hybrid population remained present, which suggests that the D. parapallidosa of hybrid origin does not represent a mere transient population but is stable. Why do these two species coexist irrespective of gene flow? We realized that body size is generally larger in D. ananassae than in D. parapallidosa, which constitutes a new character with which to discriminate these species; previously the number of sex comb teeth was the only diagnostic trait. Character displacement was not detected, however, for those traits. We crossed these two species, which resulted in offspring that had an altered genomic constitution. The body size of D. ananassae was dominant, and the presence of chromosomes Y and 4 did not have a significant effect on body size. By contrast, the presence of chromosome 4 from D. ananassae significantly affected the number of sex comb teeth. Even flies having a genomic constitution similar to that of the Penang D. parapallidosa exhibited a number of sex comb teeth that was intermediate between the two species. We propose that the D. parapallidosa sex comb character underwent selection during evolution of the Penang Island population. Reproductive interference between the species, presumably caused by signal jamming, was detected.

Evidence for variations in the morphometric traits between two sibling species of Drosophila: D. ananassae and D. pallidosa

Darwinian theory of evolution states that, evolution occurs through the natural selection. Therefore, demonstration of natural selection in nature is the central aim of many evolutionary studies and selection acts primarily at the phenotypic level because it is well known that phenotypic traits are the primary target of natural selection. While keeping this in view, we have studied certain morphometric traits in the sibling species pair, D. ananassae and D. pallidosa to test intra- and interspecific variations. The traits studied are wing length, thorax length, ratio of wing length and thorax length, sternopleural bristle number, ovariole number and sex-comb tooth number. In females of D. ananassae, significant strain differences were found for all the traits except ovariole number. In males, significant strain differences were found for all the traits. On the other hand, in D. pallidosa, significant strain differences were found for all the traits in both, males and females. The values of all the morphometric traits were significantly higher in females of both the species in comparison to males. The values of all the morphometric traits were higher in D. ananassae. However, the phenotypic variability, expressed in terms of coefficient of variation, was higher in D. pallidosa. Except for ratio of wing length and thorax length, CV was higher in the case of females in comparison to males. Size related traits are least variable while bristle numbers and reproductive traits are most variable. Except few, most of the traits are positively correlated with each other in both the species. Intra- and interspecific variations were found with respect to different morphometric traits. Although sibling species have been defined as morphologically identical, our results show that sibling species may show variations in certain morphometric traits and these quantitative differences in the morphometric traits act as discriminant marker between these sibling species in the lack of any qualitative differences.

The large X-effect on secondary sexual characters and the genetics of variation in sex comb tooth number in Drosophila subobscura.

Genetic studies of secondary sexual traits provide insights into whether and how selection drove their divergence among populations, and these studies often focus on the fraction of variation attributable to genes on the X‐chromosome. However, such studies may sometimes misinterpret the amount of variation attributable to the X‐chromosome if using only simple reciprocal F1 crosses, or they may presume sexual selection has affected the observed phenotypic variation. We examined the genetics of a secondary sexual trait, male sex comb size, in Drosophila subobscura. This species bears unusually large sex combs for its species group, and therefore, this trait may be a good candidate for having been affected by natural or sexual selection. We observed significant heritable variation in number of teeth of the distal sex comb across strains. While reciprocal F1 crosses seemed to implicate a disproportionate X‐chromosome effect, further examination in the F2 progeny showed that transgressive autosomal effects inflated the estimate of variation associated with the X‐chromosome in the F1. Instead, the X‐chromosome appears to confer the smallest contribution of all major chromosomes to the observed phenotypic variation. Further, we failed to detect effects on copulation latency or duration associated with the observed phenotypic variation. Overall, this study presents an examination of the genetics underlying segregating phenotypic variation within species and illustrates two common pitfalls associated with some past studies of the genetic basis of secondary sexual traits.

Genetic dissection of intraspecific variation in a male-specific sexual trait in Drosophila melanogaster.

An open question in evolutionary biology is the relationship between standing variation for a trait and the variation that leads to interspecific divergence. By identifying loci underlying phenotypic variation in intra- and interspecific crosses we can determine the extent to which polymorphism and divergence are controlled by the same genomic regions. Sexual traits provide abundant examples of morphological and behavioral diversity within and among species, and here we leverage variation in the Drosophila sex comb to address this question. The sex comb is an array of modified bristles or ‘teeth' present on the male forelegs of several Drosophilid species. Males use the comb to grasp females during copulation, and ablation experiments have shown that males lacking comb teeth typically fail to mate. We measured tooth number in >700 genotypes derived from a multiparental advanced-intercross population, mapping three moderate-effect loci contributing to trait heritability. Two quantitative trait loci (QTLs) coincide with previously identified intra- and interspecific sex comb QTL, but such overlap can be explained by chance alone, in part because of the broad swathes of the genome implicated by earlier, low-resolution QTL scans. Our mapped QTL regions encompass 70–124 genes, but do not include those genes known to be involved in developmental specification of the comb. Nonetheless, we identified plausible candidates within all QTL intervals, and used RNA interference to validate effects at four loci. Notably, TweedleS expression knockdown substantially reduces tooth number. The genes we highlight are strong candidates to harbor segregating, functional variants contributing to sex comb tooth number.

Interspecific hybridization does not affect the level of fluctuating asymmetry (FA) in the Drosophila bipectinata species complex.

The Drosophila bipectinata species complex comprises of four very closely related species namely D. bipectinata, D. parabipectinata, D. malerkotliana and D. pseudoananassae. It was found that irrespective of the evolutionary divergence among the species, FA which is reflective of the developmental precision remains nearly same in four species. During the present study, the level of FA in different morphological traits was studied in interspecific hybrids and compared with that of parental species with the view that it would throw light on the degree of divergence between the parental species. If they have not diverged much, the interspecific hybrids may have a similar FA level, incompatibilities between their genomes being negligible. On the other hand, if there is substantial divergence, the level of FA may be higher due to incompatibility between the genomes of the parental species. The morphological traits taken were sternopleural bristle number and wing length in both males and females and ovariole number and sex-comb tooth number in females and males respectively. However, except in a few cases, we could not detect any significant differences in the level of FA in hybrids as compared to pure species. On the other hand, a number of abnormalities like poor viability, dystrophied ovaries, asymmetrical eyes etc., could be detected in hybrids from crosses involving D. pseudoananassae as one of the parents. Therefore, we conclude that specific developmental pathways are more susceptible to developmental disturbances due to genomic incompatibilities than the large complex system bringing about developmental stability.

Studies on fluctuating asymmetry (FA) for certain morphological traits in four species of the Drosophila bipectinata complex

Fluctuating asymmetry (FA) is defined as subtle deviations from perfect bilateral symmetry, evident in differences between the right and the left sides of any given trait. It is a pattern of variation between sides and measures developmental instability. Differences in the level of FA may be used for comparing developmental precision among closely related species and thus may give an idea whether developmental stability was affected during the divergence and separation of populations into distinct species. Keeping this in view, FA was studied in four species of the Drosophila bipectinata complex i.e. D. bipectinata, D. parabipectinata, D. malerkotliana and D. pseudoananassae. In females of all the four species, FA values did not vary significantly for any of the traits considered. However, in case of males, they varied significantly for Wing length (WL) and sex comb tooth number (SCTN). Also, while in females Composite fluctuating asymmetry (CFA) did not exhibit significant variation, in males it was found to vary significantly across the four species. However, Bonferroni t-tests did not reveal any consistent difference in FA levels between any two species. The magnitude of FA was found to differ significantly among traits and CFA values were found to be higher for males than females in all the four species. Therefore, it may be concluded that the level of FA shows trait specific variations and males are more prone to developmental perturbations. However, the FA levels are more or less similar in all the four species of this complex. Thus, developmental precision remains nearly same in all the four species of this complex irrespective of the degree of evolutionary divergence reached.

Sexual selection and the evolution of secondary sexual traits: sex comb evolution in Drosophila

Sexual selection can drive rapid evolutionary change in reproductive behaviour, morphology and physiology. This often leads to the evolution of sexual dimorphism, and continued exaggerated expression of dimorphic sexual characteristics, although a variety of other alternative selection scenarios exist. Here, we examined the evolutionary significance of a rapidly evolving, sexually dimorphic trait, sex comb tooth number, in two Drosophila species. The presence of the sex comb in both D.melanogaster and D.pseudoobscura is known to be positively related to mating success, although little is yet known about the sexually selected benefits of sex comb structure. In this study, we used experimental evolution to test the idea that enhancing or eliminating sexual selection would lead to variation in sex comb tooth number. However, the results showed no effect of either enforced monogamy or elevated promiscuity on this trait. We discuss several hypotheses to explain the lack of divergence, focussing on sexually antagonistic coevolution, stabilizing selection via species recognition and nonlinear selection. We discuss how these are important, but relatively ignored, alternatives in understanding the evolution of rapidly evolving sexually dimorphic traits.

Size and symmetry of sex combs were not related to male mating success inDrosophila melanogasterreared at different temperatures

AbstractTemperature is one of the most important climatic factors that may influence different traits (morphological, physiological or behavioral) inDrosophila. In this study, we examined the effects of two developmental temperatures (18°Cand 25°C) on the size and the symmetry of sex combs (a male sexual trait) and their importance for male mating success inDrosophila melanogaster. However, the number of sex comb teeth (“size”) and its difference between right and left legs (“symmetry”) were relevant neither to male mating success nor to the growth temperatures.

Are larger and/or more symmetrical Drosophila melanogaster (Diptera, Drosophilidae) males more successful in matings in nature?

Are larger and/or more symmetrical Drosophila melanogaster (Diptera, Drosophilidae) males more successful in matings in nature? Sexual selection in Drosophila melanogaster, related to body size and fluctuating asymmetry in wing length and number of sex comb teeth in males, was tested in natural conditions. Males collected in copula were significantly larger than those collected as a single, while no difference in mean number of sex comb teeth between copulating and single males was observed. On the other hand, single males had greater asymmetry both for wing length and number of sex comb teeth than their mating counterparts. It looks like that symmetry of these bilateral traits also may play a role in sexual selection in this dipteran species in nature.

High genetic differentiation between an African and a non-African strain of Drosophila simulans revealed by segregation distortion and reduced crossover frequency

Drosophila simulans strains originating from Madagascar and nearby islands in the Indian Ocean often differ from those elsewhere in the number of sex comb teeth and the degree of morphological anomaly in hybrids with D. melanogaster. Here, we report a strong segregation distortion in the F1 intercross between two D. simulans strains originating from Madagascar and the US, possibly at both the gametic and zygotic levels. Strong bias against alleles of the Madagascar strain was observed for all ten marker loci distributed over the entire second chromosome in the F1 intercross, but only a few showed a weak distortion in the isogenic backgrounds of either strains. Significant deviations of genotype frequencies from Hardy-Weinberg proportions were consistently observed for the second chromosome. By contrast, the X and third chromosomes did not show any strong segregation distortion. Crossover frequency on the second chromosome was uniformly reduced in isogenic backgrounds whereas the map lengths in the F1 intercross were comparable to or larger than that of the standard D. melanogaster map. We discuss these findings in relation to previous studies on other traits and interspecific differences between D. mauritiana, which is endemic to Mauritius Island, and D. simulans.

Variations in morphological and life-history traits under extreme temperatures in Drosophila ananassae

Using half-sib analysis, we analysed the consequences of extreme rearing temperatures on genetic and phenotypic variations in the morphological and life-history traits of Drosophila ananassae. Paternal half-sib covariance contains a relatively small proportion of the epistatic variance and lacks the dominance variance and variance due to maternal effect, which provides more reliable estimates of additive genetic variance. Experiments were performed on a mass culture population of D. ananassae collected from Kanniyakumari (India). Two extremely stressful temperatures (18 degree C and 32 degree C) and one standard temperature (25 degree C) were used to examine the effect of stressful and non-stressful environments on the morphological and life-history traits in males and females. Mean values of various morphological traits differed signifi cantly among different temperature regimens in both males and females. Rearing at 18 degree C and 32 degree C resulted in decreased thorax length, wing-to-thorax (w/t) ratio, sternopleural bristle number, ovariole number, sex comb-tooth number and testis length. Phenotypic variances increased under stressful temperatures in comparison with non-stressful temperatures. Heritability and evolvability based on among-sires (males), among-dams (females), and the sum of the two components (sire + dam) showed higher values at both the stressful temperatures than at the non-stressful temperature. These differences reflect changes in additive genetic variance. Viability was greater at the high than the low extreme temperature. As viability is an indicator of stress, we can assume that stress was greater at 18 degree C than at 32 degree C in D. ananassae. The genetic variations for all the quantitative and life-history traits were higher at low temperature. Variation in sexual traits was more pronounced as compared with other morphometric traits, which shows that sexual traits are more prone to thermal stress. Our results agree with the hypothesis that genetic variation is increased in stressful environments.

Effect of directional selection for body size on fluctuating asymmetry in certain morphological traits in Drosophila ananassae

Variation in the subtle differences between the right and left sides of bilateral characters or fluctuating asymmetry (FA) has been considered as an indicator of an organism's ability to cope with genetic and environmental stresses during development. However, due to inconsistency in the results of empirical studies, the relationship between FA and stress has been the subject of intense debate. In this study, we investigated whether stress caused by artificial bidirectional selection for body size has any effect on the levels of FA of different morphological traits in Drosophila ananassae. The realised heritability (h2) was higher in low-line females and high-line males, which suggests an asymmetrical response to selection for body size. Further, the levels of FA were compared across 10 generations of selection in different selection lines in both sexes for sternopleural bristle number, wing length, wing-to-thorax ratio, sex combtooth number and ovariole number. The levels of FA differed significantly among generations and selection lines but did not change markedly with directional selection. However, the levels of FA were higher in the G10 generation (at the end of selection) than G0 (at the start of selection) but lower than the G5 generation in different selection lines, suggesting that the levels of FA are not affected by the inbreeding generated during the course of selection. Also, the levels of FA in the hybrids of high and low lines were signifi cantly lower than the parental selection lines, suggesting that FA is influenced by hybridisation. These results are discussed in the framework of the literature available on FA and its relationship with stress.

Fluctuating Asymmetry in Hybrids of Sibling Species, Drosophila ananassae and Drosophila pallidosa, Is Trait and Sex Specific

Due to inconsistent results of the empirical studies, the relationship between fluctuating asymmetry (FA, a measure of developmental stability) and interspecific hybridization has been the subject of intense debates. In the present study, we have assessed the impact of interspecific hybridization between 2 sibling species of Drosophila: Drosophila ananassae and Drosophila pallidosa on the levels of FA over 3 generations. Trait size of different morphological traits, namely, sternopleural bristle number, wing length (WL), wing to thorax (W/T) ratio, sex comb tooth number (SCTN), and ovariole number differed significantly among parental species and their hybrids of different generations in both the sexes. However, the levels of FA of different morphological traits were similar in parental species and their hybrids of different generations in males (except SCTN) and in females (except for WL and W/T ratio). These results are interpreted in terms of developmental stability as a function of a balance between the level of heterozygosity and the disruption of coadapted gene complexes.

Effect of Mutations on Developmental Stability and Canalization in Morphological Traits in Drosophila ananassae

The present study was designed to determine the effects of visible mutations of large effect on developmental stability and canalization in different morphological traits, namely, sternopleural bristle number, wing length, wing to thorax ratio, ovariole number, and sex comb tooth number (SCTN) in Drosophila ananassae. We have compared the mean trait size, fluctuating asymmetry (FA) (as an index of developmental stability), and morphological variation (as an index of canalization) of different mutant strains (yellow body color, y; claret eye color, ca; plexus wing, px; spread wing, spr; ebony body and sepia eye color, e se; yellow body and claret eye color, y ca; and cardinal eye color, curled wing, and ebony body color, cd cu e) with wild-type strain. The mean trait size of all morphological traits differs significantly among the wild-type and mutant strains. The wild-type and mutant strains vary significantly for the morphological variation and also for the levels of the FA in different morphological traits. However, we have found no increase in either the variance or in the degree of FA with the increase of the mutations (except in SCTN in y mutant). The plausible reasons for the variation in wild-type and mutant strains with particular reference to developmental stability and canalization have been discussed.

Effect of developmental temperature stress on fluctuating asymmetry in certain morphological traits in Drosophila ananassae

In the present study, the effect of thermal stress on the variability and fluctuating asymmetry (FA) in different morphological traits, viz., thorax length (TL), sternopleural bristle number (SBN), wing length (WL), wing-to-thorax (W/T) ratio, sex comb tooth number (SCTN) and ovariole number (ON), was investigated in 10 isofemale lines of Drosophila ananassae. The phenotypic and genetic variability is higher in the flies reared at low (20 °C) and at high (30 °C) temperatures as compared to that of standard (25 °C) temperature. Further, the levels of FA of measured traits differed significantly among the three temperature regimes except SBN and SCTN in males and SBN and W/T ratio in females. Moreover, the magnitude of positional fluctuating asymmetry is similar in males reared at three different developmental temperatures for SBN and SCTN but it varies significantly for SBN in females. However, when FA across all the traits was combined into a composite index (CFA), significant differences were found for both temperature regimes and sexes. Males showed higher CFA at 30 °C whereas in females it was higher at 20 °C. The results suggest that temperature increases the levels of variability and FA but the effect seems to be trait and sex specific in D. ananassae.

New Candidate Genes for Sex-Comb Divergence BetweenDrosophila mauritianaandDrosophila simulans

A large-effect QTL for divergence in sex-comb tooth number between Drosophila simulans and D. mauritiana was previously mapped to 73A-84AB. Here we identify genes that are likely contributors to this divergence. We first improved the mapping resolution in the 73A-84AB region using 12 introgression lines and 62 recombinant nearly isogenic lines. To further narrow the list of candidate genes, we assayed leg-specific expression and identified genes with transcript-level evolution consistent with a potential role in sex-comb divergence. Sex combs are formed on the prothoracic (front) legs, but not on the mesothoracic (middle) legs of Drosophila males. We extracted RNA from the prothoracic and mesothoracic pupal legs of two species to determine which of the genes expressed differently between leg types were also divergent for gene expression. Two good functional candidate genes, Scr and dsx, are located in one of our fine-scale QTL regions. In addition, three previously uncharacterized genes (CG15186, CG2016, and CG2791) emerged as new candidates. These genes are located in regions strongly associated with sex-comb tooth number differences and are expressed differently between leg tissues and between species. Further supporting the potential involvement of these genes in sex-comb divergence, we found a significant difference in sex-comb tooth number between co-isogenic D. melanogaster lines with and without P-element insertions at CG2791.

Fluctuating asymmetry in certain morphological traits in laboratory populations of Drosophila ananassae

Fluctuating asymmetry (FA, subtle random deviations from perfect bilateral symmetry) is often used as a measure of developmental instability (DI), which results from perturbations in developmental pathways caused by genetic or environmental stressors. During the present study, we estimated FA in 5 morphological traits, viz. wing length (WL), wing to thorax ratio (W:T), sternopleural bristle number (SBN), sex-comb tooth number (SCTN), and ovariole number (ON) in 18 laboratory populations of Drosophila ananassae. FA levels of measured traits differed significantly among populations except for SBN (in males and females) and W:T ratio (in females). Positional fluctuating asymmetry (PFA), a sensitive measure of DI, also varied significantly among the populations for SBN in females and SCTN in males. Interestingly, both males and females were similar for nonsexual traits. However, when FA across all traits (sexual and nonsexual) was combined into a single composite index (CFA), significant differences were found for both populations and sexes. Males showed higher CFA values than females, suggesting that males are more prone to developmental perturbations. The magnitude of FA differed significantly among traits, being lowest for nonsexual traits (SBN, WL, W:T ratio) and highest for sexual traits (SCTN and ON). The trait size of sexual traits (SCTN and ON) was positively correlated with their asymmetry. The possible reasons for variation in FA both among traits and among populations, and the usefulness of FA as an indicator of developmental stress and phenotypic quality in D. ananassae are discussed.

Genetic architecture of variation in sex-comb tooth number in Drosophila simulans

The sex comb on the forelegs of Drosophila males is a secondary sexual trait, and the number of teeth on these combs varies greatly within and between species. To understand the relationship between the intra- and interspecific variation, we performed quantitative trait locus (QTL) analyses of the intraspecific variation in sex-comb tooth number. We used five mapping populations derived from two inbred Drosophila simulans strains that were divergent in the number of sex-comb teeth. Although no QTLs were detected on the X chromosome, we identified four QTLs on the second chromosome and three QTLs on the third chromosome. While identification and estimated effects of the second-chromosome QTLs depend on genetic backgrounds, significant and consistent effects of the two third-chromosome QTLs were found in two genetic backgrounds. There were significant epistatic interactions between a second-chromosome QTL and a third-chromosome QTL, as well as between two second-chromosome QTLs. The third-chromosome QTLs are concordant with the locations of the QTLs responsible for the previously observed differences in sex-comb tooth number between D. simulans and D. mauritiana.

Evidence for canalization of Distal‐less function in the leg of Drosophila melanogaster

A considerable body of theory pertaining to the evolution of canalization has emerged recently, yet there have been few empirical investigations of their predictions. To address this, patterns of canalization and trait correlation were investigated under the individual and joint effects of the introgression of a loss-of-function allele of the Distal-less gene and high-temperature stress on a panel of iso-female lines. Variation was examined for number of sex comb teeth and the length of the basi-tarsus on the pro-thoracic leg of male Drosophila melanogaster. I demonstrate that whereas there is evidence for trait canalization, there is no evidence to support the hypothesis of the evolution of genetic canalization as a response to microenvironmental canalization. Furthermore, I demonstrate that although there are genetic correlations between these traits, there is no association between their measures of canalization. I discuss the prospects of the evolutionary lability of the Distal-less gene within the context of changes in genetic variation and covariation.

Genetic studies of two sister species in the Drosophila melanogaster subgroup, D. yakuba and D. santomea

We performed genetic analysis of hybrid sterility and of one morphological difference (sex-comb tooth number) on D. yakuba and D. santomea, the former species widespread in Africa and the latter endemic to the oceanic island of São Tomé, on which there is a hybrid zone. The sterility of hybrid males is due to at least three genes on the X chromosome and at least one on the Y, with the cytoplasm and large sections of the autosomes having no effect. F1 hybrid females carrying two X chromosomes from either species are perfectly fertile despite their genetic similarity to completely sterile F1 hybrid males. This implies that the appearance of Haldane's rule in this cross is at least partially due to the faster accumulation of genes causing male than female sterility. The larger effects of the X and Y chromosomes than of the autosomes, however, also suggest that the genes causing male sterility are recessive in hybrids. Some female sterility is also seen in interspecific crosses, but this does not occur between all strains. This is seen in pure-species females inseminated by heterospecific males (probably reflecting incompatibility between the sperm of one species and the female reproductive tract of the other) as well as in inseminated F1 and backcross females, probably reflecting genetically based incompatibilities in hybrids that affect the reproductive system. The latter 'innate' sterility appears to involve deleterious interactions between D. santomea chromosomes and D. yakuba cytoplasm. The difference in male sex-comb tooth number appears to involve fairly large effects of the X chromosome. We discuss the striking evolutionary parallels in the genetic basis of sterility, in the nature of sexual isolation, and in morphological differences between the D. santomea/D. yakuba divergence and two other speciation events in the D. melanogaster subgroup involving island colonization.