Genetic mapping of developmental instability: design, model and algorithm

Developmental instability or noise, defined as the phenotypic imprecision of an organism in the face of internal or external stochastic disturbances, has been thought to play an important role in shaping evolutionary processes and patterns. The genetic studies of developmental instability have been based on fluctuating asymmetry (FA) that measures random differences between the left and the right sides of bilateral traits. In this article, we frame an experimental design characterized by a spatial autocorrelation structure for determining the genetic control of developmental instability for those traits that cannot be bilaterally measured. This design allows the residual environmental variance of a quantitative trait to be dissolved into two components due to permanent and random environmental factors. The degree of developmental instability is quantified by the relative proportion of the random residual variance to the total residual variance. We formulate a mixture model to estimate and test the genetic effects of quantitative trait loci (QTL) on the developmental instability of the trait. The genetic parameters including the QTL position, the QTL effects, and spatial autocorrelations are estimated by implementing the EM algorithm within the mixture model framework. Simulation studies were performed to investigate the statistical behavior of the model. A live example for poplar trees was used to map the QTL that control root length growth and its developmental instability from cuttings in water culture.     

Tolerance assessment of Cistus ladanifer to serpentine soils by developmental stability analysis

Developmental instability is the result of random environmental perturbations during development. Its absence (developmental stability) depends on an organism's ability to buffer environmental disturbances. Both genotype and environment influence the phenotypic expression of developmental instability and it is susceptible to selection pressure. We studied developmental instability (as indicated by increased within-individual asymmetry of repeated traits) in vegetative and reproductive structures of three populations of Cistus ladanifer L. living in different soil substrates (serpentine, siliceous and contact zone) to detect tolerance to serpentine soils. Serpentine soils, characterized by high concentrations of heavy metals (Ni, Cr, and Co), low levels of Ca/Mg ratio and high water deficit, can adversely affect plant performance. In this study we demonstrated that asymmetry and within-plant variance were higher in the contact zone population than either the silica or serpentine populations, proving the adaptation of C. ladanifer to serpentine soils. Within-population estimates of developmental instability were concordant for both vegetative and reproductive traits. There was little or no within-individual correlation among estimates of developmental instability based on different structures, i.e., plants that had highly asymmetric leaves always had high developmental instability in translational symmetry. Radial asymmetry of petals was negatively correlated with petal size, especially in silica soil plants, providing evidence of selection for symmetric and large petals. While leaf size was positively correlated with absolute fluctuating asymmetry, suggesting selection for small or intermediate size leaves. Serpentine soils presented the largest foliar and floral traits, as well as shoot elongation, while silica soil plants had the smallest scores. On the contrary, aboveground plant biomass was larger in silica soil plants, while the contact zone plants had the lowest biomass.     

Are human preferences for facial symmetry focused on signals of developmental instability?

Humans find symmetrical faces more attractive than are asymmetricalfaces. Evolutionary psychologists claim that our preferencefor symmetry can be explained in the context of mate choicebecause symmetry is an honest indicator of the genetic qualityof potential mates. These arguments assume that asymmetry inhuman faces is fluctuating asymmetry (FA), because this formof asymmetry can be revealing of developmental instability.However, no study has yet examined the characteristics of facialasymmetry. Here we provide the first detailed study of the patternsof asymmetry in human faces. We measured asymmetry in 35 facialtraits. Although some traits had distributions characteristicof FA, many had distributions that characterize directionalasymmetry (DA); on average, both men and women had right hemi-facedominance. For DA traits we used deviations from the mean asymmetryas a measure of developmental instability. Our measures of asymmetryaccounted for a moderate proportion of the variance in perceivedsymmetry. Importantly, only FAs and random deviations from DAcontributed to people's perception of symmetry. DA was not importantin symmetry judgments. Faces rated as symmetrical were alsorated as attractive. Random deviations from DA were weakly relatedto women's attractiveness judgments of men's faces. DAs didnot influence attractiveness judgments. Our data suggest thatpeople focus on aspects of facial asymmetry that may be revealingof developmental instability. Further studies that isolate FAfrom other forms of asymmetry are required to accurately assessthe influence of developmental instability on the quality ofindividuals and its potential role in mate preferences.     

Leaf herbivory and fluctuating asymmetry as indicators of mangrove stress

Wetlands Ecology and Management - Fluctuating asymmetry (FA), a widely used measure of developmental instability in plants and animals, which describes random differences in size and/or shape...     

The effect of nutrient stress on developmental instability in leaves of Acer platanoides (Aceraceae) and Betula pendula (Betulaceae)

Studies of developmental stability can provide insights into the amount of genetic or environmental stress experienced by individuals or populations. In the present study, we used young plants of Acer platanoides (Norway maple) and Betula pendula (silver birch), two distantly related tree species with widely different leaf morphologies, to compare the expression of developmental instability in two contrasting environments: one with free access to nutrients and the other with a severely limited supply of nutrients. Using the difference in size between the right and left side of each leaf as a measure of developmental instability, we found no effect of nutrient deficiency on leaf asymmetry, despite large sample sizes (370-380 plants per species and treatment) and evidence for stress-related changes in overall leaf size and plant biomass. Moreover, there was no consistent relationship between individual leaf asymmetry and plant biomass within each nutrient treatment. In view of these observations, leaf asymmetry appears to be a poor indicator of nutrient stress in young plants of Acer platanoides and Betula pendula.     

Developmental instability and immune function in colour polymorphic pygmy grasshoppers

Random left minus right deviations from symmetry in otherwise bilaterally symmetric traits may arise due to developmental instability in response to environmental stress. Here we test for variation in developmental instability, measured as asymmetry of (femur) size, among individuals belonging to four different genetically encoded colour morphs of the pygmy grasshopper Tetrix undulata (Sow.) (Orthoptera: Tetrigidae). Such a difference is expected under the hypotheses that perturbed growth and development and concomitant morphological asymmetry may result from exposure to unfavourable temperatures or costs associated with melanization, or reflect a by-product of past disease in individuals with poor immune responsiveness. Results from mixed model two-way ANOVAs uncovered no statistically significant directional asymmetry in femur size, whereas non-directional asymmetry was significant in each of the four different colour morphs. The degree of intraindividual, interlateral variance in femur size varied significantly among individuals belonging to different colour morphs, being considerably higher in the black morph, which also suffers the greatest risk of overheating, compared to the brown, striped and grey morphs. Asymmetry in femur size was not associated with one measure of immune responsiveness, the encapsulation response of individuals experimentally implanted with a novel antigen consisting of a nylon mono-filament. These results are consistent with the notion that individual pigmentation pattern may indirectly influence developmental instability and morphological asymmetry, via the effects of coloration on body temperature.Co-ordinating editor: Hurst     

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.     

Fruit abortion,developmental selection and developmental stability in<Emphasis Type="Italic"> Quercus ilex</Emphasis>

Fruit abortion has been hypothesized to be a parental means of selective removal of propagules with low viability. In particular, aborted zygotes have been suggested to have developmentally deviant phenotypes, and surviving offspring may therefore give rise to adults with a developmentally stable phenotype. We tested predictions from this hypothesis using acorns of holm oaks Quercus ilex as a model system. Fecundity of oak trees was negatively related to mean fluctuating asymmetry of leaves, and abortion rates were positively related to leaf fluctuating asymmetry in at least one population. Aborted acorns were asymmetric in 83-99% of cases in three samples, while mature acorns were only asymmetric in 57-78% of cases. Acorn asymmetry was unrelated to germination probability and germination date, and had no significant effect on number of leaves, leaf mass, stem mass, seedling height or leaf area of seedlings. However, acorn asymmetry affected the trade-off between number and size of leaves in seedlings. Seedlings from asymmetric acorns showed a positive relationship between acorn size and number of leaves, but no relationships between acorn size and leaf area, while symmetric acorns showed the opposite. A positive relationship between acorn size and number of leaves in spring was found for naturally emerged seedlings that died during their first summer, whereas the number of leaves produced by surviving seedlings did not depend on acorn size. These findings are consistent with the hypothesis that stressed trees selectively abort propagules of low viability, and that developmental selection acts on a measure of developmental instability of fruits.     

Developmental Instability and Sexual Selection in Stag Beetles from Chernobyl and a Control Area

Radioactivity may have both phenotypic and genetic effects through its disruption of physiological processes and mutations. I studied the size and asymmetry of secondary sexual and ordinary morphological characters of stag beetles ( Lucanus cervus ) in two areas in the Ukraine: near Chernobyl, where levels of radiation are high, and in a control area with low background radiation. Developmental instability of morphological characters was estimated from the degree of fluctuating asymmetry using the restricted maximum likelihood parameter estimation (REML) method to partition measurement error from asymmetry. The degree of asymmetry estimated from unsigned differences in size of right and left secondary sexual character was similar to estimates based on the REML method. Beetles from the contaminated area had a significantly elevated level of fluctuating asymmetry in the secondary sexual character compared with animals from the control area. Male stag beetles found with a female had significantly lower asymmetry than males found alone. While mated males did not differ in asymmetry between areas, unmated males from Chernobyl were much more asymmetric than unmated males from the control area. These findings provide evidence for radiation disrupting developmental homeostasis, and thereby affecting the mating status of free-living beetles.     

Symmetry breaking in interspecific Drosophila hybrids is not due to developmental noise

Hybrids from crosses of different species have been reported to display decreased developmental stability when compared to their pure species, which is conventionally attributed to a breakdown of coadapted gene complexes. Drosophila subobscura and its close relative D. madeirensis were hybridized in the laboratory to test the hypothesis that genuine fluctuating asymmetry, measured as the within-individual variance between right and left wings that results from random perturbations in development, would significantly increase after interspecific hybridization. When sires of D. subobscura were mated to heterospecific females following a hybrid half-sib breeding design, F1 hybrid females showed a large bilateral asymmetry with a substantial proportion of individuals having an asymmetric index larger than 5% of total wing size. Such an anomaly, however, cannot be plainly explained by an increase of developmental instability in hybrids but is the result of some aberrant developmental processes. Our findings suggest that interspecific hybrids are as able as their parents to buffer developmental noise, notwithstanding the fact that their proper bilateral development can be harshly compromised. Together with the low correspondence between the co-variation structures of the interindividual genetic components and the within-individual ones from a Procrustes analysis, our data also suggest that the underlying processes that control (genetic) canalization and developmental stability do not share a common mechanism. We argue that the conventional account of decreased developmental stability in interspecific hybrids needs to be reappraised.     

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.     

Geometric morphometrics and leaf phenotypic plasticity: assessing fluctuating asymmetry and allometry in European white oaks (Quercus)

Because plants are unable to move away from unfavourable habitats and environmental perturbations, leaf phenotypic plasticity facilitates light absorption and gas exchange. Oaks (Quercus spp.) are particularly known for their adaptability and plastic phenotypes, and leaf allometry and developmental instability may represent important mechanisms for their adaptation to environments and evolution. Because of its important role in the adaptation of plant populations to different environments, allometry can be involved in diversifying selection. Developmental instability is related to environmental perturbations and stresses by producing random deviations in structures characterized by bilateral symmetry, such as oak leaves. In addition, developmental instability can also arise from genetic bottlenecks or as a result of hybridization. The splitting of symmetric and asymmetric components of variation and their separate analysis allows the variability in leaf shape traits to be summarized, reducing the variation produced by developmental instability. The geometric morphometric approach is a useful method for the study of leaf asymmetry and allometric patterns. This method provides an important tool for the visualization of shape attributes that characterize species with highly variable leaf phenotypic patterns. In this study, leaf shape and size variability of three white oak species was investigated by means of a two‐dimensional landmark‐based method providing improved knowledge of variance partitioning, species discrimination, fluctuating asymmetry and allometric patterns of variation resulting from the different analyses. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 335–348.     

A latent variable model of developmental instability in relation to men's sexual behaviour

A single trait's fluctuating asymmetry (FA) is expected to be a poor measure of developmental instability. Hence, studies that examine associations between FA and outcomes expected to covary with developmental instability often have little power in detecting meaningful relationships. One way of increasing the power of detecting relationships between developmental instability and outcomes is through the use of multiple traits' FA. The way multiple traits have typically been used is in trait aggregates. Here, we illustrate another way of examining relationships with developmental instability using multiple traits' FA: through structural equation modelling. Covariances between measures of FA and an outcome variable are interpreted within the context of an explicit model of associations between variables, which is tested for fit and the parameters specified within the model are estimated. We used nine traits' FA as markers of a latent variable of men's developmental instability, which was associated with the number of sexual partners. The results indicate a sizeable correlation between developmental instability and men's sexual history, despite small correlations between individual traits' FA and sexual history.     

Asymmetric size and shape variation in the Central European transect across the house mouse hybrid zone

We studied asymmetric variation of the mandible in the Central European portion of the hybrid zone between two house mouse subspecies, Mus musculus musculus and Mus musculus domesticus. Within introgression classes, defined by the share of diagnostic allozymes, we quantified the directional and fluctuating component of asymmetric variation, as well as skewness and kurtosis of individual asymmetry distributions. Furthermore, in the same manner we re‐analysed asymmetric variation of the ventral side of the skull. According to the quadratic polynomial model, the mandible shape‐fluctuating asymmetry, but not size‐fluctuating asymmetry, was significantly decreased in the centre of the hybrid zone (with a minimum predicted for a hybrid index of 0.41). On the contrary, the skull shape‐fluctuating asymmetry non‐monotonically increased towards the musculus side of the hybrid zone (with a peak predicted for a hybrid index of 0.86). Thus, the impact of hybridization on fluctuating asymmetry is trait‐specific in this portion of the house mouse hybrid zone. The only general feature of asymmetric variation we observed was the shift towards the platykurtosis of asymmetry distributions in the centre of the hybrid zone. Taken together, we suggest genetic variability for right–left asymmetries to be generally increased, but the developmental instability of mandible shape to be decreased, by hybridization. We hypothesize the decrease of developmental instability to be caused by overdominant effects on developmental dynamics rather than by increased heterozygosity. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 13–27.     

The repeatability of fluctuating asymmetry: a revision and extension

Fluctuating asymmetry (FA) is an imperfect measure of the developmental stability of an individual. Estimates of the heritability of FA will underestimate the heritability of developmental stability; the extent of this bias can be estimated and corrected by estimating the repeatability of developmental stability. This note corrects an error in a previous derivation of this repeatability using the absolute value of the difference between sides as the measure of FA, and derives the repeatability of developmental stability for another common measure of FA, the variance among sides within an individual.     

Does isolation affect phenotypic variability and fluctuating asymmetry in the endangered Red Apollo?

Isolated populations represent one of the main focuses in conservation biology. Long-term isolation often causes losses of genetic diversity and as a consequence might reduce individual fitness. Morphometric characters can be used as suitable markers to analyse ecological stress and individual fitness of local populations. Asymmetry in bilateral symmetry is used as a measure for developmental instability of populations and is often negatively correlated with population size and low genetic diversity. As a study system, we selected the endangered butterfly Parnassius apollo, which occurs in small and isolated remnant populations in Central Europe, but also in fairly large metapopulations in the Alps. We analysed wing morphometrics (shape and size characters) of 812 individuals representing (1) already extinct, (2) highly isolated and (3) still interconnected populations. Seventeen landmarks on veins were used to analyse morphological variances in the wing shape. Our data show significant deviations between landmarks on the left and right wing side within individuals and strong morphological variance among individuals. The highest morphological variability could be found for individuals in the Alps, however, the level of asymmetry was very similar for all populations analysed. The higher morphological variability found in the Alps can be interpreted as a consequence of the higher level of the genetic diversity detectable for this region. Analysis on morphological variance of P. apollo individuals of the Mosel valley using time series ranging from 1895 until today showed no significant rise in asymmetry and no decline of morphological variability over time, although, anthropogenic habitat destruction has caused severe bottlenecks in this population.     

The Functional Significance of Chiral Genitalia: Patterns of Asymmetry,Functional Morphology and Mating Success in the Praying Mantis Ciulfina baldersoni

Genital asymmetry is relatively common and widespread throughout the animal kingdom. The functional significance of genital asymmetry is however, poorly understood for most species. Male praying mantids of the genus Ciulfina are remarkable in possessing complex and directionally asymmetric genital phallomeres in some species, and chirally dimorphic/antisymmetric genitalia in others. Here we explore the chiral dimorphism in male genitalia of Ciulfina baldersoni which appear to exhibit genital antisymmetry. We test whether genital orientation influences mating success, copulation duration and the attachment duration of spermatophores. Additionally we investigate genital interactions between male and females using x-ray micro-computed tomography (micro-CT) and scanning electron microscopy (SEM). Lastly we assess whether genital asymmetry is associated with non-genital morphological asymmetry of a range of traits. Our results highlight the complex functional morphology of genitalia in this praying mantis species and yet demonstrate no functional difference between dextral and sinistral morphs other than the direction of attachment with both morphs enjoying equal levels of mating success. Chiral morphs also did not strongly associate with any other forms of asymmetry. We therefore conclude that genital chirality in Ciulfina baldersoni is a likely case of antisymmetry with no functional significance to genital orientation, and is likely to be selectively neutral.     

Antisymmetry,directional asymmetry,and dynamic morphogenesis

Fluctuating asymmetry is the most commonly used measure of developmental instability. Some authors have claimed that antisymmetry and directional asymmetry may have a significant genetic basis, thereby rendering these forms of asymmetry useless for studies of developmental instability. Using a modified Rashevsky-Turing reaction-diffusion model of morphogenesis, we show that both antisymmetry and directional asymmetry can arise from symmetry-breaking phase transitions. Concentrations of morphogen on right and left sides can be induced to undergo transitions from phase-locked periodicity, to phase-lagged periodicity, to chaos, by simply changing the levels of feedback and inhibition in the model. The chaotic attractor has two basins of attraction-right sidedominance and left side dominance. With minor disturbance, a developmental trajectory settles into one basin or the other. With increasing disturbance, the trajectory can jump from basin to basin. The changes that lead to phase transitions and chaos are those expected to occur with either genetic change or stress. If we assume that the morphogen influences the behavior of cell populations, then a transition from phase-locked periodicity to chaos in the morphogen produces a corresponding transition from fluctuating asymmetry to antisymmetry in both morphogen concentrations and cell populations. Directional asymmetry is easily modeled by introducing a bias in the conditions of the simulation. We discuss the implications of this model for researchers using fluctuating asymmetry as an indicator of stress.     

2,3,7,8-Tetrachlorodibenzo-p-dioxin affects fluctuating asymmetry of molar shape in mice, and an epistatic interaction of two genes for molar size

Fluctuating asymmetry (FA), random variation between left and right sides in a bilaterally symmetrical character, is a commonly used measure of developmental instability that is expected to increase with increasing environmental stress. One potential stressor is 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a powerful toxicant known to disturb tooth development. In this study, mice in the F(2) generation produced from an intercross between two inbred strains (C57BL/6J and AKR/J) were exposed in utero to TCDD. We hypothesized that TCDD would increase FA in the molars of exposed mice over that of the control mice. In addition, we hypothesized that we would discover genes for molar size, shape or asymmetry whose expression would be affected by TCDD. We detected a very small, but significant, increase in FA of molar shape (but not size) in the TCDD-exposed mice compared to the control mice, although molar size and shape did not differ between these groups. Although we did not uncover any genes that acted differently in the TCDD exposed and control groups, we did identify two genes whose dominance by additive epistatic effect on molar size was affected by TCDD. We concluded that although TCDD may be affecting the expression of some genes governing the development of molars in our population of mice, FA of molar size and shape is not a particularly sensitive indicator of this effect.     

Fluctuating asymmetry and stress in a medieval Nubian population

Fluctuating asymmetry is commonly used as a bioindicator of developmental stress. This study addresses asymmetry under nutritional/systemic stress in the human craniofacial skeleton and its utility as an indicator of developmental instability. Crania from the diachronic Christian cemeteries at Kulubnarti (Sudanese Nubia) were chosen as a model for nutrition/systemic stress. Previous studies indicate that individuals from the Early Christian cemetery were subjected to greater developmental stress when compared with individuals from the Late Christian cemetery. Therefore, crania from the Early Christian cemetery should display a greater magnitude of fluctuating asymmetry than crania from the Late Christian cemetery. Thirty adult crania of comparable age and sex were selected from each population. Landmark coordinates were digitized in two separate trials and averaged to minimize error. Euclidean distance matrix analysis (EDMA) was used to measure and compare the magnitude of fluctuating asymmetry in each sample. Results indicate that crania from the Early Christian cemetery display greater amounts of fluctuating asymmetry than those from the Late Christian cemetery, as predicted. The degree of fluctuating asymmetry for each linear distance is highly correlated between the cemeteries, suggesting that all humans may share common patterns of fluctuating asymmetry in the skull. In contrast, there is little correlation between magnitude of fluctuating asymmetry and length of linear distance, between-subject variability, or measurement error. These results support the hypothesis that poor nutrition/systemic stress increases developmental instability in the human skull and that increased fluctuating asymmetry constitutes morphological evidence of this stress.