Natural genetic variation in fluctuating asymmetry of wing shape in Drosophila melanogaster

Fluctuating asymmetry (FA), defined as random deviation from perfect symmetry, has been used to assay the inability of individuals to buffer their developmental processes from environmental perturbations (i.e., developmental instability). In this study, we aimed to characterize the natural genetic variation in FA of wing shape in Drosophila melanogaster, collected from across the Japanese archipelago. We quantified wing shapes at whole wing and partial wing component levels and evaluated their mean and FA. We also estimated the heritability of the mean and FA of these traits. We found significant natural genetic variation in all the mean wing traits and in FA of one of the partial wing components. Heritability estimates for mean wing shapes were significant in two and four out of five wing traits in males and females, respectively. On the contrary, heritability estimates for FA were low and not significant. This is a novel study of natural genetic variation in FA of wing shape. Our findings suggest that partial wing components behave as distinct units of selection for FA, and local adaptation of the mechanisms to stabilize developmental processes occur in nature.     

Temperature and genotypic effects on life history and fluctuating asymmetry in a field strain of Culex pipiens

Fluctuating asymmetry (FA) has been proposed as a tool to measure levels of stress experienced by populations of organisms during development. To be of value as a bio-marker to highlight conditions at particular sites, it is important that variation in FA is due to environmental (eg pollution) variation and not genetic variation among populations and families, in other words heritability for FA should be very close to zero. A full-sib design was set up in which families of Culex pipiens mosquitoes collected from the field were reared at three different developmental temperatures. The effects of temperature and family on developmental rate, egg to adult survival and four wing morphological measures were assessed. There was both a temperature and a family effect on development rate and survival. Temperature affected all four wing traits, but an influence of family was only evident in two of the wing traits. Two separate measures of FA for each of the wing traits were obtained. The mean estimates of FA were mainly around 1% of the value of the character measured. There was evidence of an increase in FA with increase in temperature stress. Heritability was estimated for the wing traits and wing trait FA's using restricted estimation maximum likelihood. The estimates of heritability for the wing traits were small and, individually, did not differ significantly from zero. There was also no evidence of heritable genetic variation for any of the wing trait FA's. The results are discussed in relation to other studies where FA heritabilities have been estimated and in relation to the use of FA as an indicator of environmental stress.     

Artificial selection reveals heritable variation for developmental instability

Fluctuating (nondirectional) asymmetry (FA) of bilaterally paired structures on a symmetrical organism is commonly used to assay the developmental instability (DI) caused by environmental or genetic factors. Although evidence for natural selection to reduce FA has been reported, evidence that FA (and by extension DI) is heritable is weak. We report the use of artificial selection to demonstrate heritable variation in the fluctuating asymmetry of interlandmark distances within the wing in an outbred population of Drosophila melanogaster. Our estimates for the heritability of FA range from 0% to 1% and result in estimates for the heritability of DI as large as 20%, comparable to values typical for life-history traits. These values indicate the existence of evolutionarily relevant genetic variation for DI and the effectiveness of selection for reduced FA suggests that natural selection has not fixed all the genetic variants that would improve developmental stability in these populations.     

Genetic architecture of wing morphology in Drosophila simulans and an analysis of temperature effects on genetic parameter estimates

The Drosophila wing has been used as a model to investigate the mechanisms responsible for size and shape changes in nature, since such changes might underlie morphological evolution. To improve the understanding of wing morphological variation and the interpretation of genetic parameters estimates, we have established 59 lines from a Drosophila simulans laboratory population through single pair random matings. The offspring of each line were reared at three different temperatures, and the wing morphology of 12 individuals was analyzed by adjusting an ellipse to the wings' contour. Temperature, sex and line significantly affected wing trait variation, which was mainly characterized by longer wings having the second, fourth and fifth longitudinal veins closer together at the wing tip. As for the genetic parameter estimates, while the cross-environment heritability of some traits, such as wing size (SI), decreased with an increasing difference between the temperatures at which parents and offspring were reared, wing shape (SH) heritability did not seem to change. Since we found indications that neither an increase in the phenotypic variation nor the occurrence of genotype-environment interactions could fully explain the low heritabilities of SI estimated by cross-environment regressions, we discuss the importance of other effects for explaining this discrepancy between the SI and SH heritability estimates. In addition, although the genetic matrix was not entirely represented in the phenotypic matrix, several correspondences were identified, suggesting that the observed patterns of wing morphology variation are genetically controlled.     

Heritability of Wing Morphology in a Natural Population of Drosophila gouveai

The natural and laboratory heritabilities of a series of parameters related to wing size and shape were estimated in a population of Drosophila gouveai (repleta group) under field and laboratory conditions. A morphometric analysis was done using 17 wing parameters related to wing landmark positions obtained using the method of the best adjustment of an ellipse to the wing edge. Three parameters (thetaA, thetaC and thetaD) showed highly significant heritability in the wild (average 0.61), whereas only wing size (W(SI)) had significant heritability in the laboratory (0.71). The additive genetic variance of most parameters was greater in the wild than in the laboratory. These results showed that some parameters possessed a substantial genetic additive component in their phenotypic variance, and that morphometric parameters of D. gouveai wings are appropriate quantitative markers for assessing morphological differentiation among populations.     

Effects of parental radiation exposure on developmental instability in grasshoppers

Mutagenic and epigenetic effects of environmental stressors and their transgenerational consequences are of interest to evolutionary biologists because they can amplify natural genetic variation. We studied the effect of parental exposure to radioactive contamination on offspring development in lesser marsh grasshopper Chorthippus albomarginatus. We used a geometric morphometric approach to measure fluctuating asymmetry (FA), wing shape and wing size. We measured time to sexual maturity to check whether parental exposure to radiation influenced offspring developmental trajectory and tested effects of radiation on hatching success and parental fecundity. Wings were larger in early maturing individuals born to parents from high radiation sites compared to early maturing individuals from low radiation sites. As time to sexual maturity increased, wing size decreased but more sharply in individuals from high radiation sites. Radiation exposure did not significantly affect FA or shape in wings nor did it significantly affect hatching success and fecundity. Overall, parental radiation exposure can adversely affect offspring development and fitness depending on developmental trajectories although the cause of this effect remains unclear. We suggest more direct measures of fitness and the inclusion of replication in future studies to help further our understanding of the relationship between developmental instability, fitness and environmental stress.     

Structural Characteristics of Allomyrina Dichotoma Beetle＇s Hind Wings for Flapping Wing Micro Air Vehicle

In this study, we present a complete structural analysis ofAllomyrina dichotoma beetle＇s hind wings by investigating their static and dynamic characteristics. The wing was subjected to the static loading to determine its overall flexural stiffness. Dy- namic characteristics such as natural frequency, mode shape, and damping ratio of vibration modes in the operating frequency range were determined using a Bruel ＆ Kjaer fast Fourier transform analyzer along with a laser sensor. The static and dynamic characteristics of natural Allomyrina dichotoma beetle＇s hind wings were compared to those of a fabricated artificial wing. The results indicate that natural frequencies of the natural wing were significantly correlated to the wing surface area density that was defined as the wing mass divided by the hind wing surface area. Moreover, the bending behaviors of the natural wing and artificial wing were similar to that of a cantilever beam. Furthermore, the flexural stiffness of the artificial wing was a little higher than that of the natural one whereas the natural frequency of the natural wing was close to that of the artificial wing. These results provide important information for the biomimetic design of insect-scale artificial wings, with which highly ma- neuverable and efficient micro air vehicles can be designed.     

PLASTICITY, CANALIZATION, AND DEVELOPMENTAL STABILITY OF THE DROSOPHILA WING: JOINT EFFECTS OF MUTATIONS AND DEVELOPMENTAL TEMPERATURE

The phenotypic effects of genetic and environmental manipulations have been rarely investigated simultaneously. In addition to phenotypic plasticity, their effect on the amount and directions of genetic and phenotypic variation is of particular evolutionary importance because these constitute the material for natural selection. Here, we used heterozygous insertional mutations of 16 genes involved in the formation of the Drosophila wing. The flies were raised at two developmental temperatures (18°C and 28°C). Landmark-based geometric morphometrics was used to analyze the variation of the wing size and shape at different hierarchical levels: among genotypes and temperatures; among individuals within group; and fluctuating asymmetry (FA). Our results show that (1) the phenotypic effects of the mutations depend on temperature; (2) reciprocally, most mutations affect wing plasticity; (3) both temperature and mutations modify the levels of FA and of among individuals variation within lines. Remarkably, the patterns of shape FA seem unaffected by temperature whereas those associated with individual variation are systematically altered. By modifying the direction of available phenotypic variation, temperature might thus directly affect the potential for further evolution. It suggests as well that the developmental processes responsible for developmental stability and environmental canalization might be partially distinct.     

Quantitative-genetic analysis of wing form and bilateral asymmetry in isochromosomal lines ofDrosophila subobscura using Procrustes methods

Fluctuating asymmetry (FA) is often used as a measure of underlying developmental instability (DI), motivated by the idea that morphological variance is maladaptive. Whether or not DI has evolutionary potential is a highly disputed topic, marred by methodological problems and fuzzy prejudices. We report here some results from an ongoing study of the effects of karyotype, homozygosity and temperature on wing form and bilateral asymmetry using isochromosomal lines ofDrosophila subobscura. Our approach uses the recently developed methodologies in geometric morphometrics to analyse shape configurations of landmarks within the standard statistical framework employed in studies of bilateral asymmetries, and we have extended these methods to partition the individual variation and the variation in asymmetries into genetic and environmental causal components. The analyses revealed temperaturedependent expression of genetic variation for wing size and wing shape, directional asymmetry (DA) of wing size, increased asymmetries at suboptimal temperature, and a transition from FA to DA in males as a result of increase in the rearing temperature. No genetic variation was generally detected for FA in our samples, but these are preliminary results because no crosses between lines were carried out and, therefore, the contribution of dominance was not taken into account. In addition, only a subset of the standing genetic variation was represented in the experiments.     

The effects of latitude,body size,and sexual selection on wing shape in a damselfly

Under natural selection, wing shape is expected to evolve to optimize flight performance. However, other selective factors besides flight performance may influence wing shape. One such factor could be sexual selection in wing sexual ornaments, which may lead to alternative variations in wing shape that are not necessarily related to flight performance. In the present study, we investigated wing shape variations in a calopterygid damselfly along a latitudinal gradient using geometric morphometrics. Both sexes show wing pigmentation, which is a known signal trait at intra‐ and interspecific levels. Wing shape differed between sexes and, within the same sex, the shape of the hind wing differed from the front wing. Latitude and body size explained a high percentage of the variation in wing shape for female front and hind wings, and male front wings. In male hind wings, wing pigmentation explained a high amount of the variation in wing shape. On the other hand, the variation in shape explained by pigmentation was very low in females. We suggest that the conservative morphology of front wings is maintained by natural selection operating on flight performance, whereas the sex‐specific differences in hind wings most likely could be explained by sexual selection. The observed sexual dimorphism in wing shape is likely a result of different sex‐specific behaviours. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102 , 263–274.     

Genetic variation for the positioning of wing veins in Drosophila melanogaster

SUMMARY To define the components of variation for wing shape in Drosophila in relation to what is known about the developmental control of wing patterning, we have characterized shape variation in the wings of 12 randomly chosen highly inbred lines. Despite large differences in wing size between males and females, and between flies reared at 18°C or 25°C, wing shape is remarkably unaffected by these variables and is highly line specific. The shape of each intervein region of the wing appears to be independently regulated at the genetic level, consistent with the role of secreted growth factors in establishing the locations of wing veins. Sex and temperature were found to have different effects on cell number in two intervein regions, with the result that wing shape is to a large extent independent of cell density. Dietary cholesterol was also shown to affect the breadth of the central intervein region, consistent with an effect on the strength of Hedgehog signaling during wing development. We conclude that wing shape is under tighter genetic control than wing size, and hypothesize that this control is achieved in large part by gene activity at the level of wing vein determination and differentiation.     

Does stress mess with rodents’ heads? Influence of habitat amount and genetic factors in mandible fluctuating asymmetry in South American water rats (Nectomys squamipes,Sigmodontinae) from Brazilian Atlantic rainforest remnants

Loss of developmental stability can lead to deviations from bilateral symmetry (i.e. Fluctuating Asymmetry ‐ FA), and is thought to be caused by environmental and genetic factors associated with habitat loss and stress. Therefore, levels of FA might be a valuable tool to monitor wild populations if FA serves as an indicator of exposure to stress due to impacts of habitat loss and fragmentation. In studies examining FA and habitat fragmentation, FA levels are often explained by loss of genetic variation, though few studies have addressed FA’s use as indicator of environmental impact. Here, we investigated whether habitat loss, genetic variation, and/or inbreeding affect the developmental instability in Brazilian Atlantic forest populations of a Neotropical water rat (Nectomys squamipes). We sampled individuals from eight sites within Atlantic forest remnants with different amounts of available forest habitat and assessed FA levels with geometric morphometric techniques using adult mandibles. We used observed heterozygosity (H_o) and inbreeding coefficient (F_is), from seven microsatellite markers, as a proxy of genetic variation at individual and population levels. Populations were not significantly different for shape or size FA levels. Furthermore, interindividual variation in both shape and size FA levels and interpopulational differences in size FA levels were best explained by chance. However, habitat amount was negatively associated with both interpopulational variance and average shape FA levels. This association was stronger in populations living in areas with <28% of forest cover, which presented higher variance and higher average FA, suggesting that Nectomys squamipes might have a tolerance threshold to small availability of habitat. Our work is one of the first to use FA to address environmental stress caused by habitat loss in small mammal populations from a Neotropical biome. We suggest that shape FA might serve as a conservation tool to monitor human impact on natural animal populations.     

Rhodnius prolixus and Rhodnius robustus–like (Hemiptera, Reduviidae) wing asymmetry under controlled conditions of population density and feeding frequency

Habitat change in Rhodnius spp may represent an environmental challenge for the development of the species, particularly when feeding frequency and population density vary in nature. To estimate the effect of these variables in stability on development, the degree of directional asymmetry (DA) and fluctuating asymmetry (FA) in the wing size and shape of R. prolixus and R. robustus–like were measured under laboratory controlled conditions. DA and FA in wing size and shape were significant in both species, but their variation patterns showed both inter-specific and sexual dimorphic differences in FA of wing size and shape induced by nutrition stress. These results suggest different abilities of the genotypes and sexes of two sylvatic and domestic genotypes of Rhodnius to buffer these stress conditions. However, both species showed non-significant differences in the levels of FA between treatments that simulated sylvan vs domestic conditions, indicating that the developmental noise did not explain the variation in wing size and shape found in previous studies. Thus, this result confirm that the variation in wing size and shape in response to treatments constitute a plastic response of these genotypes to population density and feeding frequency.     

Wing shape heritability and morphological divergence of the sibling species Drosophila mercatorum and Drosophila paranaensis

The fruit-flies Drosophila paranaensis and Drosophila mercatorum pararepleta are sibling species belonging to the repleta group. Females of these two species are normally considered to be morphologically indistinguishable while males only differ consistently in the morphology of their genitalia. These species are sympatric throughout a large area of their geographic distribution. In this study, we investigated the degree of morphological divergence between D. paranaensis and D. mercatorum pararepleta based on morphometric analysis of their wings. The ellipse method was used to describe the placement of the longitudinal and transversal wing veins as well as the size of the wing and the shape of its outline. The heritability under laboratory and field conditions was also estimated from the parameters generated. Multivariate analysis showed that wing morphology possessed sufficient differences to discriminate between the two species with a successful classification rate of 95-98% for females and 82-87% for males. The results of the autoclassification were confirmed by a cross-validation test for females (92-96%). Most measurements possessed significant natural heritability (a mean of 0.48 for D. mercatorum and 0.88 for D. paranaensis), indicating that the variation observed was related to differences in genes acting additively. The principal difference between the two species was in the placement of the posterior transverse wing vein. However, the pattern of morphological variation in the wings of both species was similar, possibly because of shared restrictions in wing development pathways.     

Geometric estimates of heritability in biological shape

The recently developed geometric morphometrics methods represent an important contribution of statistics and geometry to the study of biological shapes. We propose simple protocols using shape distances that incorporate geometric techniques into linear quantitative genetic models that should provide insights into the contribution of genetics to shape variation in organisms. The geometric approaches use Procrustes distances in a curved shape space and distances in tangent spaces within and among families to estimate shape heritability. We illustrate the protocols with an example of wing shape variation in the honeybee, Apis mellifera. The heritability of overall shape variation was small, but some localized components depicting shape changes on distal wing regions showed medium to large heritabilities. The genetic variance-covariance matrix of the geometric shape variables was significantly correlated with the phenotypic shape variance-covariance matrix. A comparison of the results of geometric methods with the traditional multivariate analysis of interlandmark distances indicated that even with a larger dimensionality, the interlandmark distances were not as rich in shape information as the landmark coordinates. Quantitative genetics studies of shape should greatly benefit from the application of geometric methods.     

Morphological intergration between development compartments in the Drosophila wing

Abstract. Developmental integration is the covariation among morphological structures due to connections between the developmental processes that built them. Here we use the methods of geometric morphometrics to study integration in the wing of Drosophila melanogaster . In particular, we focus on the hypothesis that the anterior and posterior wing compartments are separate developmental units that vary independently. We measured both variation among genetically diverse individuals and random differences between body sides of single individuals (fluctuating asymmetry, FA). For both of these sources of variation, the patterns of variation identified by principal component analyses all involved landmarks in both the anterior and posterior compartments simultaneously. Analyses focusing exclusively on the covariation between the anterior and posterior compartments, by the partial least-squares method, revealed pervasive integration of the two compartments, for both individual variation and FA. These analyses clearly indicate that the anterior and posterior compartments are not separate units of variation, but that the covariation between compartments is sufficient to account for nearly all the variation throughout the entire wing. We conclude that variation among individuals as well as the developmental perturbations responsible for FA generate shape variation primarily through developmental processes that are integrated across both compartments. In contrast, much less of the shape variation in our sample can be attributed to the localized processes that establish the identity of particular wing veins.     

Phenotypic Buffering in a Monogenean: Canalization and Developmental Stability in Shape and Size of the Haptoral Anchors of Ligophorus cephali (Monogenea: Dactylogyridae)

Phenotypic variation results from the balance between sources of variation and counteracting regulatory mechanisms. Canalization and developmental stability are two such mechanisms, acting at two different levels of regulation. The issue of whether or not they act concurrently as a common developmental buffering capacity has been subject to debate. We used geometric morphometrics to quantify the mechanisms that guarantee phenotypic constancy in the haptoral anchors of Ligophorus cephali. Canalization and developmental stability were appraised by estimating inter- and intra-individual variation, respectively, in size and shape of dorsal and ventral anchors. The latter variation was estimated as fluctuating asymmetry (FA) between anchor pairs. The general-buffering-capacity hypothesis was tested by two different methods based on correlations and Principal Components Analyses of the different components of size and shape variation. Evidence for FA in the dorsal and ventral anchors in both shape and size was found. Our analyses supported the hypothesis of a general developmental buffering capacity. The evidence was more compelling for shape than for size and, particularly, for the ventral anchors than for the dorsal ones. These results are in line with previous studies of dactylogyrids suggesting that ventral anchors secure a firmer, more permanent attachment, whereas dorsal anchors are more mobile. Because fixation to the host is crucial for survival in ectoparasites, we suggest that homeostatic development of the ventral anchors has been promoted to ensure the morphological constancy required for efficient attachment. Geometric morphometrics can be readily applied to other host-monogenean models, affording not only to disentangle the effects of canalization and developmental stability, as shown herein, but to further partition the environmental and genetic components of the former.     

Detecting genetic variation in developmental instability by artificial selection on fluctuating asymmetry

Abstract Fluctuating asymmetry (FA) is frequently used as a measure of developmental instability (DI). Assuming a genetic basis to DI, many have argued that FA may be a good indicator of genetic quality to potential mates and to human managers of populations. Unfortunately FA is a poor indicator of DI, making it very difficult to verify this assertion. A recent review of the literature suggests that previous studies of the inheritance of FA and DI using half‐sib covariances and parent–offspring regression have been unable to put meaningful limits on the heritability of FA and DI because of the extremely low power of the experiments performed. In this study, we consider the power of artificial selection on FA as an alternative approach to studying the inheritance of FA and DI. Using simulations, we investigate the efficacy of selection for both increased and decreased FA for detecting genetic variation. We find that selection for increased FA has much more power to detect the presence of genetic variation than does selection for decreased FA. These results hold when realistic sample sizes are employed. Artificial selection for increased FA is currently the most powerful approach for the detection of genetic variation in DI.     

Evolution of the male genitalia: morphological variation of the aedeagi in a natural population of <Emphasis Type="Italic">Drosophila mediopunctata</Emphasis>

To investigate the size and shape of the aedeagus of Drosophila mediopunctata, we used basic statistics and geometric morphometrics. We estimated the level of phenotypic variation, natural and laboratory heritability as well as the phenotypic correlations between aedeagus and wing measures. The wing was used as an indicator for both body size and shape. Positive significant correlation was obtained for centroid size of aedeagus and wing for field parents and their offspring reared in the laboratory. Many positive significant phenotypic correlations were found among linear measures of both organs. The phenotypic correlations were few for aedeagus and wing shape. Coefficients of variation of the measures were on average larger in the aedeagus than in the wing for offspring reared in laboratory, but not for flies coming from the field. Significant “natural” heritabilities were found for five linear measures of the aedeagus and only one for the wing. Few significant heritabilities were found for aedeagus and wing shape, mostly ones concerning the uniform components. In an exploratory analysis, we found that inversion DS-PC0 is associated with both uniform and nonuniform components of shape, respectively, in the wing and aedeagus. Our results do not support the lock-and-key hypothesis for the male genitalia evolution, but cannot refute the sexual selection and pleiotropy hypotheses.     

Stability of Development and Variability in Morphological Traits in a Natural Population of Drosophila melanogaster: Seasonal Dynamics in 1999

Seasonal dynamics of developmental stability and variability of morphological traits was examined in a natural population of Drosophila melanogaster in order to compare these two parameters as indicators of temperature stress. Morphometric (thorax length and wing length) and meristic (number of sternopelural and orbital bristles) were studied. Variability was measured as the coefficient of variation. Stability of development was estimated as fluctuating asymmetry (FA) of bilateral traits. Thorax length and wing length did not exhibit consistent seasonal trends whereas wing loading significantly decreased. Significant seasonal changes in FA were not detected in any trait examined. Two traits showed reduced variation in autumn. The use of FA as an indicator of ecological stresses in insects is discussed on the basis of these results and the literature evidence.