The phenotypic correlates and quantitative genetics of masculinization in the rodent, Octodon degus.

This corrects the article DOI: 10.1038/hdy.2017.20.

The phenotypic correlates and quantitative genetics of masculinization in the rodent, Octodon degus.

In some mammals, female characteristics have been shown to depend, in part, on the intrauterine position during development of female fetuses relative to male fetuses. Females developing in close proximity to males show behavioral, physiological and life history characteristics that are masculinized. With the exception of one inconclusive study, nothing is known of the genetic basis of this phenomenon. In this paper, we reported an analysis of the quantitative genetic basis of masculinization, as indicated by the anogenital distance (AGD) at birth and weaning, in the rodent Octodon degus. Because AGD is related to weight, we included a genetic analysis of pup weight at birth and weaning. Pairwise correlations showed that AGD at birth varied negatively with litter size and parturition number but positively with weaning AGD, birth weight, dam AGD and percentage of males in the litter. AGD at weaning varied similarly except that it tended to vary positively with litter size. Genetic (co)variances of AGD at birth and weight at birth differed in females and males. In females, the best genetic model included substantial effects of direct additive, additive maternal and a negative additive genetic covariance between these two. In males, variances were small and there was difficulty in discriminating between additive maternal and common environmental variances. By weaning, genetic (co)variances had somewhat declined in weight and were not statistically significant in AGD in either sex. This paper showed the occurrence of both phenotypic and genetic components in masculinization with effects being greater in females.

Intra- and trans-generational effects of larval diet on susceptibility to an entomopathogenic fungus, Beauveria bassiana, in the greater wax moth, Galleria mellonella.

In addition to nutritional conditions experienced by individuals themselves, those experienced by their parents can affect their immune function. Here, we studied the intra- and trans-generational effects of larval diet on susceptibility to an entomopathogenic fungus, Beauveria bassiana, in the greater wax moth, Galleria mellonella. In the first part of the study, a split-brood design was used to compare the susceptibility of full sibs raised either on low- or on high-nutrition larval diet. In the second part of the study, a similar experimental design was employed to investigate the effects of maternal and paternal diet as well as their interaction on offspring's susceptibility. In the first part of the study, we found that individuals fed with high-nutrition diet had higher mortality from infection than individuals fed with low-nutrition diet. However, diet did not affect post-infection survival time. Conversely, in the second part of the study, maternal diet was found to have no significant effect on final mortality rate of offspring, but it affected survival time: larvae with high-nutrition maternal diet survived fewer days after infection than larvae with low-nutrition maternal diet. Paternal diet had no significant effect on offspring's susceptibility to the fungus, indicating that paternal effects are not as important as maternal effects in influencing immune function in this species. Our findings provide further indication that maternal nutrition affects immune function in insects, and suggest that the direct effects of nutrition on immunity may be different, yet parallel, to those caused by parental nutrition.

Bias in the heritability of preference and its potential impact on the evolution of mate choice.

The evolution of mate choice is a function of the heritability of preference. Estimation in the laboratory is typically made by presenting a female with a limited number of males. We show that such an approach produces a downwardly biased estimate, which we term the heritability of choice. When preference is treated as a threshold trait then less biased estimates are obtained particularly for preferences based on the relative value of the preferred trait. Because females in the wild typically survey on average less than five males we argue that the heritability of choice may be more meaningful than the heritability of preference. The restricted number of males surveyed can lead to a reduction in the phenotypic variance of the preferred trait in the group of males selected by the females if the phenotypic variance in preference is equal to or less than the phenotypic variance in the referred trait. If the phenotypic variance in preference exceeds that of the preferred trait then the opposite occurs. A second effect of the restricted number of males sampled is that females are likely to mate initially with males that are not the most preferred. The failure to find the most preferred male may account for the common observation of multiple matings and extra-pair copulations. We suggest that current explanations for polyandry need to take this failure into account.

The quantitative genetics of sexually selected traits, preferred traits and preference: a review and analysis of the data.

The maintenance of variation in sexually selected traits is a puzzle that has received increasing attention in the past several decades. Traits that are related to fitness, such as life-history or sexually selected traits, are expected to have low additive genetic variance (and hence, heritability) due to the rapid fixation of advantageous alleles. However, previous analyses have suggested that the heritabilities of sexually selected traits are on average higher than nonsexually selected traits. We show that the heritabilities of sexually selected traits are not significantly different from those of nonsexually selected traits overall or when separated into the three trait categories: behavioural, morphological and physiological. In contrast with previous findings, the heritability of preference is quite low (h(2) = 0.25 ± 0.06) and is in the same range as life-history traits. We distinguish preferred traits as a category of sexually selected traits and find that the heritability of the former is not significantly different than sexually selected traits overall (0.48 ± 0.04 vs. 0.46 ± 0.03). We test the hypothesis that the heritability of sexually selected traits is negatively correlated with the strength of sexual selection. As predicted, there is a significant negative correlation between the heritabilities of sexually selected traits and the strength of selection. This suggests that heritabilities do indeed decrease as sexual selection increases but sexual selection is not strong enough to cause heritabilities of sexually selected traits to deviate from the same type of nonsexually selected traits.

The costs of being dark: the genetic basis of melanism and its association with fitness-related traits in the sand cricket.

Melanism is an important component of insect cuticle and serves numerous functions that enhance fitness. Despite its importance, there is little information on its genetic basis or its phenotypic and genetic correlation with fitness-related traits. Here, we examine the heritability of melanism in the wing dimorphic sand cricket and determine its phenotypic and genetic correlation with wing morphology, gonad mass and size of the dorso-longitudinal muscles (the principle flight muscles). Previously demonstrated trade-offs among these traits are significant factors in the evolution of life history variation. Using path analysis, we show that melanization is causally related to gonad mass, but not flight muscle mass. Averaged over the sexes, the heritability of melanism was 0.61, the genetic correlation with gonad mass was -0.36 and with wing morph was 0.51. The path model correctly predicted the ranking of melanization score in lines selected for increased ovary mass, increased flight muscle mass, an index that increased both traits and an unselected control. Our results support the general hypothesis that melanization is costly for insects and negatively impacts investment in early reproduction.

Genetic and phenotypic relationships between immune defense, melanism and life-history traits at different temperatures and sexes in Tenebrio molitor.

Insect cuticle melanism is linked to a number of life-history traits, and a positive relationship is hypothesized between melanism and the strength of immune defense. In this study, the phenotypic and genetic relationships between cuticular melanization, innate immune defense, individual development time and body size were studied in the mealworm beetle (Tenebrio molitor) using three different temperatures with a half-sib breeding design. Both innate immune defense and cuticle darkness were higher in females than males, and a positive correlation between the traits was found at the lowest temperature. The effect of temperature on all the measured traits was strong, with encapsulation ability and development time decreasing and cuticle darkness increasing with a rise in temperature, and body size showing a curved response. The analysis showed a highly integrated system sensitive to environmental change involving physiological, morphological and life-history traits.

There is more than one way to skin a G matrix.

Because of its importance in directing evolutionary trajectories, there has been considerable interest in comparing variation among genetic variance-covariance (G) matrices. Numerous statistical approaches have been suggested but no general analysis of the relationship among these methods has previously been published. In this study, we used data from a half-sib experiment and simulations to explore the results of applying eight tests (T method, modified Mantel test, Bartlett's test, Flury hierarchy, jackknife-manova, jackknife-eigenvalue test, random skewers, selection skewers). Whereas a randomization approach produced acceptable estimates, those from a bootstrap were typically unacceptable and we recommend randomization as the preferred method. All methods except the jackknife-eigenvalue test gave similar results although a fine-scale analysis suggested that the former group can be subdivided into two or possibly three groups, hierarchical tests, skewers and the rest (jackknife-manova, modified Mantel, T method, probably Bartlett's). An advantage of the jackknife methods is that they permit tests of association with other factors, such as in this case, temperature and sex. We recommend applying all the tests described in this article, with the exception of the T method, and provide R functions for this purpose.

Path analysis of the genetic integration of traits in the sand cricket: a novel use of BLUPs.

This study combines path analysis with quantitative genetics to analyse a key life history trade-off in the cricket, Gryllus firmus. We develop a path model connecting five traits associated with the trade-off between flight capability and reproduction and test this model using phenotypic data and estimates of breeding values (best linear unbiased predictors) from a half-sibling experiment. Strong support by both types of data validates our causal model and indicates concordance between the phenotypic and genetic expression of the trade-off. Comparisons of the trade-off between sexes and wing morphs reveal that these discrete phenotypes are not genetically independent and that the evolutionary trajectories of the two wing morphs are more tightly constrained to covary than those of the two sexes. Our results illustrate the benefits of combining a quantitative genetic analysis, which examines statistical correlations between traits, with a path model that focuses upon the causal components of variation.

Analysis of the effects of early nutritional environment on inbreeding depression in Drosophila melanogaster.

The impact of nutritional deficiencies early in life in determining life-history variation in organisms is well recognized. The negative effects of inbreeding on fitness are also well known. Contrary to studies on vertebrates, studies on invertebrates are not consistent with the observation that inbreeding compromises resistance to parasites and pathogens. In this study, we investigated the effect of early nutrition on the magnitude of inbreeding depression in development time, adult body size and adult resistance to the bacterium Serratia marcescens in Drosophila melanogaster. We found that early nutritional environment had no effect on the magnitude of inbreeding depression in development time or adult body size but may have played a small role in adult resistance to the bacterial infection. Estimates of heritabilities for development time under the poor nutritional environment were larger than those measured under the standard nutritional conditions.

Trade-off acquisition and allocation in Gryllus firmus: a test of the Y model.

Many models of life history evolution assume trade-offs between major life history traits; however, these trade-offs are often not found. The Y model predicts that variation in acquisition can mask underlying allocation trade-offs and is a major hypothesis explaining why negative relationships are not always found between traits that are predicted to trade-off with one another. Despite this model's influence on the field of life history evolution, it has rarely been properly tested. We use a model system, the wing dimorphic cricket, Gryllus firmus as a case study to test the assumptions and predictions of the Y model. By experimentally altering the acquisition regime and by estimating energy acquisition and energy allocation directly in this species, we are able to explicitly test this important model. Overall, we find strong support for the predictions of the Y model.

Genetic independence of female signal form and male receiver design in the almond moth, Cadra cautella.

Efficient signalling requires coordination of signal form and receiver design. To maintain signal function, parallel changes in signaller and receiver traits are required. Genetic correlation and co-evolution among signal and response traits have been proposed to preserve signal function (i.e. coordination) during the evolution of mate recognition systems. Empirical studies have provided support for both mechanisms; however, there is debate regarding the interpretation of some of these studies. Tests for a genetic correlation typically hybridize divergent signalling systems and look at hybrid signal form and receiver design, or impose artificial selection on signal form and look for an indirect response to selection in receiver design. Some of the hybridization studies did not achieve reassortment of genes from the parental types, whereas some of the artificial selection studies incorporated random mating in their designs. As a result of these limitations, the hybridization studies cannot discriminate between genetic correlation and co-evolution with primarily additive genetic effects underlying signal and response traits. Similarly, the artificial selection experiments cannot discriminate between genetic correlation because of linkage disequilibrium and co-evolution. This study examined the mating preferences of male almond moths, Cadra cautella, before and after female moths were artificially selected (using a design incorporating assortative mating) for novel pheromone blend ratios. Our results demonstrate the absence of a genetic correlation between signal and response traits in the almond moth.

Male sand crickets trade-off flight capability for reproductive potential.

In this paper, we test the hypothesis that male sand crickets, Gryllus firmus, experience a trade-off between flight capability and reproductive potential expressed as reduced testis weight in flight-capable morphs. We used a half-sib design with 130 sires, three dams per sire and an average of 5.66 males per dam family, for a total of 2206 F1 offspring. Traits measured were head width, somatic dry weight, testis weight, wing morph (micropterous/macropterous), weight of the dorso-longitudinal flight muscles (DLM) and the functional status of these muscles. Heritabilities of all traits were significant and ranged from 0.14 to 0.43. All traits were positively correlated with body size, but removal of this covariance revealed a highly significant trade-off, both phenotypically and genetically, between testes size and flight capability as measured by wing morph, DLM size or DLM status. The possible implications of this for morph-specific reproductive tactics are discussed.

Comparing sire and dam estimates of heritability: jackknife and likelihood approaches.

Three estimates of heritability are available from the half-sib pedigree design: the sire, dam and genotypic estimates. Because of its significantly smaller standard error, the genotypic estimate is preferred provided that there are no non-additive effects that inflate the estimate. I present two methods to test for such effects: these are a t-test of the paired sire and dam pseudovalues from the jackknife procedure and the likelihood ratio test from the animal model. Both methods are shown to be valid tests for significant dominance and/or maternal effects. SPLUS coding for the implementation of the jackknife method is provided. Unless sample sizes are very large, the power of the tests is low and hence caution is advised in the use of the genotypic estimate following a nonsignificant test. An approximate power analysis can be done using the data from the jackknife method but the estimated power is typically a substantial underestimate of the true power and its use is not recommended.

Inbreeding and extreme outbreeding cause sex differences in immune defence and life history traits in Epirrita autumnata.

Empirical studies in vertebrates support the hypothesis that inbreeding reduces resistance against parasites and pathogens. However, studies in insects have not found any evidence that inbreeding compromises immune defence. Here we tested whether one generation of brother-sister mating or extreme outbreeding (mating between two populations) have an effect on innate immunity and life history traits in the autumnal moth, Epirrita autumnata. We show that the effect of inbreeding on immune response differed between the sexes: whereas in females, inbreeding significantly reduced encapsulation response against nylon monofilament ability, it did not have a significant effect on male immune response. There were also differences in the correlation of the immune response with other traits: in females increased immune response was positively correlated with large size, whereas in males immune response increased with a reduction in development time. Immune response differed significantly among families in males but not in females, both for the inbreeding and extreme outbreeding experiments. In conjunction with the observed immune responses to inbreeding, these data suggest that in males genetic variation for immune response is largely additive or non-directional with respect to dominance, whereas in females variation is much reduced and consists of directional dominance variance. Further, we show that encapsulation response against nylon monofilament is associated with the resistance against real pathogens suggesting that this widely used method to measure the strength of immune defence in insects is also a biologically relevant method.

The evolution of trade-offs: where are we?

Trade-offs are a core component of many evolutionary models, particularly those dealing with the evolution of life histories. In the present paper, we identify four topics of key importance for studies of the evolutionary biology of trade-offs. First, we consider the underlying concept of 'constraint'. We conclude that this term is typically used too vaguely and suggest that 'constraint' in the sense of a bias should be clearly distinguished from 'constraint' in the sense of proscribed combinations of traits or evolutionary trajectories. Secondly, we address the utility of the acquisition-allocation model (the 'Y-model'). We find that, whereas this model and its derivatives have provided new insights, a misunderstanding of the pivotal equation has led to incorrect predictions and faulty tests. Thirdly, we ask how trade-offs are expected to evolve under directional selection. A quantitative genetic model predicts that, under weak or short-term selection, the intercept will change but the slope will remain constant. Two empirical tests support this prediction but these are based on comparisons of geographic populations: more direct tests will come from artificial selection experiments. Finally, we discuss what maintains variation in trade-offs noting that at present little attention has been given to this question. We distinguish between phenotypic and genetic variation and suggest that the latter is most in need of explanation. We suggest that four factors deserving investigation are mutation-selection balance, antagonistic pleiotropy, correlational selection and spatio-temporal variation, but as in the other areas of research on trade-offs, empirical generalizations are impeded by lack of data. Although this lack is discouraging, we suggest that it provides a rich ground for further study and the integration of many disciplines, including the emerging field of genomics.

The importance of growth and mortality costs in the evolution of the optimal life history.

A central assumption of life history theory is that the evolution of the component traits is determined in part by trade-offs between these traits. Whereas the existence of such trade-offs has been well demonstrated, the relative importance of these remains unclear. In this paper we use optimality theory to test the hypothesis that the trade-off between present and future fecundity induced by the costs of continued growth is a sufficient explanation for the optimal age at first reproduction, alpha, and the optimal allocation to reproduction, G, in 38 populations of perch and Arctic char. This hypothesis is rejected for both traits and we conclude that this trade-off, by itself, is an insufficient explanation for the observed values of alpha and G. Similarly, a fitness function that assumes a mortality cost to reproduction but no growth cost cannot account for the observed values of alpha. In contrast, under the assumption that fitness is maximized, the observed life histories can be accounted for by the joint action of trade-offs between growth and reproductive allocation and between mortality and reproductive allocation (Individual Juvenile Mortality model). Although the ability of the growth/mortality model to fit the data does not prove that this is the mechanism driving the evolution of the optimal age at first reproduction and allocation to reproduction, the fit does demonstrate that the hypothesis is consistent with the data and hence cannot at this time be rejected. We also examine two simpler versions of this model, one in which adult mortality is a constant proportion of juvenile mortality [Proportional Juvenile Mortality (PJM) model] and one in which the proportionality is constant within but not necessarily between species [Specific Juvenile Mortality (SSJM) model]. We find that the PJM model is unacceptable but that the SSJM model produces fits suggesting that, within the two species studied, juvenile mortality is proportional to adult mortality but the value differs between the two species.

A quantitative analysis of the mechanism that controls body size in Manduca sexta.

BACKGROUND: Body size is controlled by mechanisms that terminate growth when the individual reaches a species-specific size. In insects, it is a pulse of ecdysone at the end of larval life that causes the larva to stop feeding and growing and initiate metamorphosis. Body size is a quantitative trait, so it is important that the problem of control of body size be analyzed quantitatively. The processes that control the timing of ecdysone secretion in larvae of the moth Manduca sexta are sufficiently well understood that they can be described in a rigorous manner. RESULTS: We develop a quantitative description of the empirical data on body size determination that accurately predicts body size for diverse genetic strains. We show that body size is fully determined by three fundamental parameters: the growth rate, the critical weight (which signals the initiation of juvenile hormone breakdown), and the interval between the critical weight and the secretion of ecdysone. All three parameters are easily measured and differ between genetic strains and environmental conditions. The mathematical description we develop can be used to explain how variables such as growth rate, nutrition, and temperature affect body size. CONCLUSION: Our analysis shows that there is no single locus of control of body size, but that body size is a system property that depends on interactions among the underlying determinants of the three fundamental parameters. A deeper mechanistic understanding of body size will be obtained by research aimed at uncovering the molecular mechanisms that give these three parameters their particular quantitative values.

The quantitative genetics of sexual dimorphism: assessing the importance of sex-linkage.

Sexual dimorphism (SD) is a defining feature of gonochorous animals and dioecious plants, but the evolution of SD from an initially monomorphic genome presents a conundrum. Theory predicts that the evolution of SD will be facilitated if genes with sex-specific fitness effects occur on sex chromosomes. We review this theory and show that it generates three testable predictions. For organisms with an XX/XY chromosomal system of sex determination: (1) SD should be associated with X-linked effects; (2) X-linked effects should show strong directional dominance for sexually dimorphic traits favored in males but expressed in both sexes; and (3) SD should be associated with a reduction in the between-sex additive genetic covariance and correlation. A literature review reveals that empirical evaluations of the association between sex-linkage and SD have lagged behind theory. Tests for the presence of sex-linked effects have been plagued by the need to make simplifying assumptions, such as the absence of dominance or maternal effects, that greatly weaken their discriminatory power. Further, most have used comparisons between species or populations, whereas the correct level of analysis is within populations. To overcome these problems, we derive a novel pedigree design that permits separate estimation of X-linked, dominance and maternal effects. We suggest that the data from such a design would be most appropriately analyzed using the animal model. This novel protocol will allow quantitative evaluation of the above predictions, and hence should spur progress in understanding the role of sex-linkage in the evolution of SD.

Analysis of the importance of genotypic variation, metabolic rate, morphology, sex and development time on immune function in the cricket, Gryllus firmus.

Immune defence is hypothesized to be a trait that bears significant fitness costs as well as benefits in that mounting a defence depreciates the value of other life-history traits. Thus the cost of mounting an immune response could affect the evolution of both the immune system and correlated life history traits. In this study we examined, by means of a diallel cross of four inbred lines, the genetic basis of two measures of immune function, metabolic rate and several traits in the sand cricket, Gryllus firmus. We specifically addressed the following questions: (1) is immune function determined primarily by genetic constitution or correlations with phenotypic traits that could reduce the effectiveness of the immune response; (2) do the two measures of immune function covary; (3) What are the contributions of additive, nonadditive and maternal effects to the immune function? As estimates of immune function, we used lytic activity and encapsulation rate. We found that inbred crickets were smaller than individuals from the crossed lines and took longer to develop. However, inbred lines did not differ from the crossed lines in immune function nor metabolic rates, suggesting that increased homozygosity has little or no effect on these traits in G. firmus. We found that both immune parameters showed significant genetic variation but no consistent relationships with the other phenotypic traits (metabolic rate, head width, body mass, development time and activity). There was significant additive genetic variation only in encapsulation rate, but, with the exception of the activity measure, significant nonadditive and reciprocal variances were found in all traits. Metabolic rate of crickets was heritable, but there was neither phenotypic nor genetic association between metabolic rate and the two parameters of immune function. Further, there was no correlation between these two measures. Females showed a higher encapsulation response than males, but there was no sex differences in lytic activity. Our study indicates that genetic variation in immune parameters can be a very significant contributor to phenotypic variation in immune function.