Age-specific Effects of Novel Mutations in Drosophila Melanogaster I. Mortality

Theories for the evolution of aging rest on the assumption that at least some deleterious mutations have effects that are limited to certain ages. Many mutation accumulation studies have tried to measure the number and magnitude of deleterious mutations, but few studies have tried to determine the extent to which the effects of mutations are limited to particular ages. Here we estimate the age-specific effect of deleterious mutations on mortality rate in an outbred population of the fruit fly, Drosophila melanogaster. We used the ‘middle class neighborhood’ approach to accumulation mutations in populations of flies that had recently been obtained from the wild. There are mutations that increase mortality rates, but whose effects are limited to specific ages. The age-specificity of mutational effects differs between the sexes, between virgin and mated flies, and over time. After 10 and 20 generations of mutation accumulation, there were clear age-specific effects of mutations. After 30 generations, however, the degree of age-specificity decreased. In addition, mutation accumulation led to a steady increase in larval mortality and a small but significant increase in the sex ratio of eclosing flies. We discuss the implications of these results for models of aging, and suggest approaches that future studies should take to obtain accurate information on the age-specificity of novel mutations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Heterosis in age‐specific selected populations of a seed beetle: Sex differences in longevity and reproductive behavior

We tested mutation accumulation hypothesis for the evolution of senescence using short‐lived and long‐lived populations of the seed‐feeding beetle, Acanthoscelides obtectus (Say), obtained by selection on early‐ and late‐life for many generations. The expected consequence of the mutation accumulation hypothesis is that in short‐lived populations, where the force of natural selection is the strongest early in life, the late‐life fitness traits should decline due to genetic drift which increases the frequency of mutations with deleterious effects in later adult stages. Since it is unlikely that identical deleterious mutations will increase in several independent populations, hybrid vigor for late‐life fitness is expected in offspring obtained in crosses among populations selected for early‐life fitness traits. We tested longevity of both sexes, female fecundity and male reproductive behavior for hybrid vigor by comparing hybrid and nonhybrid short‐lived populations. Hybrid vigor was confirmed for male virility, mating speed and copulation duration, and longevity of both sexes at late ages. In contrast to males, the results on female fecundity in short‐lived populations did not support mutation accumulation as a genetic mechanism for the evolution of this trait. Contrary to the prediction of this hypothesis, male mating ability indices and female fecundity in long‐lived populations exhibited hybrid vigor at all assayed age classes. We demonstrate that nonhybrid long‐lived populations diverged randomly regarding female and male reproductive fitness, indicating that sexually antagonistic selection, when accompanied with genetic drift for female fecundity and male virility, might be responsible for overriding natural selection in the independently evolving long‐lived populations.     

Probing the evolution of senescence inDrosophila melanogaster withP-element tagging

Natural populations host a wealth of genetic variation in longevity and age-specific schedules of reproduction. This variation provides critical information for inferring the evolutionary origin of senescence. Patterns of mutational effects on age-specific fecundity and survival provide additional insight to distinguish alternative models of senescence. In this study,P-elements bearing thewhite minigene were inserted at random into a common genetic background, generating lines ofD. melanogaster with single, stable transposon inserts. A series of 48 single-P-element lines revealed statistically significant heterogeneity in both longevity and fecundity. Longevity and early fecundity were only weakly positively correlated (r=0.286,P=0.0398). Both the pooled sample and 30 of the individual lines exhibited a leveling of age-specific mortality at advanced ages, in opposition to the classical demographic models. To the extent that these mutational effects are representative of naturally-occurring mutations in heterogeneous populations, this result presents a problem for the evolutionary theory of senescence. Natural selection is inefficient at removing deleterious mutations that are expressed only at late ages, and selection may not differentiate between mutations whose effects on longevity are post-reproductive. A leveling of the mortality rate would also be seen if mutations whose expression is delayed until very late simply do not occur. A simulation of mutation-selection balance among the 48P-element tagged lines shows that the mean longevity declines monotonically with increasing mutation rate, consistent with the mutation-accumulation model.     

Effect of rate of inbreeding on inbreeding depression in Drosophila melanogaster

Summary This experiment was designed to study the relationship between rate of inbreeding and observed inbreeding depression of larval viability, adult fecundity and cold shock mortality in Drosophila melanogaster. Rates of inbreeding used were full-sib mating and closed lines of N=4 and N=20. Eight generations of mating in the N=20 lines, three generations in the N=4 lines and one generation of full-sib mating were synchronised to simultaneously produce individuals with an expected level of inbreeding coefficient (F) of approximately 0.25. Inbreeding depression for the three traits was significant at F=0.25. N=20 lines showed significantly less inbreeding depression than full-sib mated lines for larval viability at approximately the same level of F. A similar trend was observed for fecundity. No effect of rate of inbreeding depression was found for cold shock mortality, but this trait was measured with less precision than the other two. Natural selection acting on loci influencing larval viability and fecundity during the process of inbreeding could explain these results. Selection is expected to be more effective with slow rates of inbreeding because there are more generations and greater opportunity for selection to act before F=0.25 is reached. Selection intensities seem to have been different in the three traits measured. Selection was most intense for larval viability, less intense for fecundity and, perhaps, negligible at loci influencing cold shock mortality.     

Spontaneous Mutational Variances and Covariances for Fitness-Related Traits in Drosophila Melanogaster

Starting from a completely homozygous population of Drosophila melanogaster, 176 lines were derived and independently maintained by a single brother-sister mating per generation. Three fitness-related traits were considered (fecundity, egg-to-pupa and pupa-to-adult viabilities). Mutational heritabilities of these traits and genetic correlations between all possible pairs were calculated from the between line divergence (codivergence), after 104-106 generations of mutation accumulation. Mutational heritabilities ranged from 0.60 X 10(-3) to 0.82 X 10(-3) and correlations from -0.11 to 0.25. These values are likely to be underestimates due to selection against deleterious mutations. The distribution of the means of the lines was asymmetric, positive for fecundity and negative for both viability components. The coefficients of asymmetry are also likely to be biased, again due to selection. Extreme lines from the two tails of the distribution were examined in detail. Homozygous line effects were all negative for viability traits but predominantly positive for fecundity, indicating the fixation of mutations with positive effects on the latter. Corresponding heterozygous line effects showed a variable degree of dominance.     

POLYGENIC MUTATION IN DROSOPHILA MELANOGASTER: ESTIMATES FROM DIVERGENCE AMONG INBRED STRAINS

A highly inbred line of Drosophila melanogaster was subdivided into 25 replicate sublines, which were independently maintained for 100 generations with 10 pairs of unselected flies per generation. The polygenic mutation rate (V_M) for two quantitative traits, abdominal and sternopleural bristle number, was estimated from divergence among sublines at 10 generation intervals from generations 30-100, and from response of each line to divergent selection after more than 65 generations of mutation accumulation. Estimates of V_M averaged over males and females both from divergence among lines and from response to selection within lines were 3.3 × 10^-3 V_E for abdominal bristles and 1.5 × 10^-3 V_E for sternopleural bristles, where V_E is the environmental variance. The actual rate of production of mutations affecting these traits may be considerably higher if the traits are under stabilizing selection, and if mutations affecting bristle number have deleterious effects on fitness. There was a substantial component of variance for sex × mutant effect interaction and the sublines evolved highly significant mutational variation in sex dimorphism of abdominal bristle number. Pleiotropic effects on sex dimorphism may be a general property of mutations at loci determining bristle number.     

The Effects of Spontaneous Mutation on Quantitative Traits. I. Variances and Covariances of Life History Traits

We have accumulated spontaneous mutations in the absence of natural selection in Drosophila melanogaster by backcrossing 200 heterozygous replicates of a single high fitness second chromosome to a balancer stock for 44 generations. At generations 33 and 44 of accumulation, we extracted samples of chromosomes and assayed their homozygous performance for female fecundity early and late in adult life, male and female longevity, male mating ability early and late in adult life, productivity (a measure of fecundity times viability) and body weight. The variance among lines increased significantly for all traits except male mating ability and weight. The rate of increase in variance was similar to that found in previous studies of egg-to-adult viability, when calculated relative to trait means. The mutational correlations among traits were all strongly positive. Many correlations were significantly different from 0, while none was significantly different from 1. These data suggest that the mutation-accumulation hypothesis is not a sufficient explanation for the evolution of senescence in D. melanogaster. Mutation-selection balance does seem adequate to explain a substantial proportion of the additive genetic variance for fecundity and longevity.     

The evolution of age-specific mortality rates in Drosophila melanogaster: genetic divergence among unselected lines.

Age-specific effects of spontaneous mutations on mortality rates in Drosophila are inferred from three large demographic experiments. Data were collected from inbred lines that were allowed to accumulate spontaneous mutations for 10, 19, and 47 generations. Estimates of age-specific mutational variance for mortality were based on data from all three experiments, totalling approximately 225,000 flies, using a model developed for genetic analysis of age-dependent traits (the character process model). Both within- and among-generation analyses suggest that the input of genetic variance is greater for early life mortality rates than for mortality at older ages. In females, age-specific mutational variances ranged over an order of magnitude from 5.96 x 10(-3) at 2 wk posteclosion to 0.02 x 10(-3) at 7 wk. The male data show a similar pattern. Age-specific genetic variances were substantially less at generation 47 than at generation 19-an unexplained observation that is likely due to block effects. Mutational correlations among mortality rates at different ages tend to increase with the accumulation of new mutations. Comparison of the mutation-accumulation lines at generations 19 and 47 with their respective control lines suggests little age-specific mutational bias.     

Age-specific properties of spontaneous mutations affecting mortality in Drosophila melanogaster.

An analysis of the effects of spontaneous mutations affecting age-specific mortality was conducted using 29 lines of Drosophila melanogaster that had accumulated spontaneous mutations for 19 generations. Divergence among the lines was used to estimate the mutational variance for weekly mortality rates and the covariance between weekly mortality rates at different ages. Significant mutational variance was observed in both males and females early in life (up to approximately 30 days of age). Mutational variance was not significantly different from zero for mortality rates at older ages. Mutational correlations between ages separated by 1 or 2 wk were generally positive, but they declined monotonically with increasing separation such that mutational effects on early-age mortality were uncorrelated with effects at later ages. Analyses of individual lines revealed several instances of mutation-induced changes in mortality over a limited range of ages. Significant age-specific effects of mutations were identified in early and middle ages, but surprisingly, mortality rates at older ages were essentially unaffected by the accumulation procedure. Our results provide strong evidence for the existence of a class of polygenic mutations that affect mortality rates on an age-specific basis. The patterns of mutational effects measured here relate directly to recently published estimates of standing genetic variance for mortality in Drosophila, and they support mutation accumulation as a viable mechanism for the evolution of senescence.     

The effect of spontaneous mutations on competitive ability

Understanding the impact of spontaneous mutations on fitness has many theoretical and practical applications in biology. Although mutational effects on individual morphological or life‐history characters have been measured in several classic genetic model systems, there are few estimates of the rate of decline due to mutation for complex fitness traits. Here, we estimate the effects of mutation on competitive ability, an important complex fitness trait, in a model system for ecological and evolutionary genomics, Daphnia. Competition assays were performed to compare fitness between mutation‐accumulation (MA) lines and control lines from eight different genotypes from two populations of Daphnia pulicaria after 30 and 65 generations of mutation accumulation. Our results show a fitness decline among MA lines relative to controls as expected, but highlight the influence of genomic background on this effect. In addition, in some assays, MA lines outperform controls providing insight into the frequency of beneficial mutations.     

MORTALITY PLATEAUS AND THE EVOLUTION OF SENESCENCE: WHY ARE OLD-AGE MORTALITY RATES SO LOW?

Age-specific mortality rates level off far below 100% at advanced ages in experimental populations of Drosophila melanogaster and other organisms. This observation is inconsistent with the equilibrium predictions of both the antagonistic pleiotropy and mutation accumulation models of senescence, which, under a wide variety of assumptions, predict a “wall” of mortality rates near 100% at postreproductive ages. Previous models of age-specific mortality patterns are discussed in light of recent demographic data concerning late-age mortality deceleration and age-specific properties of new mutations. The most recent theory (Mueller and Rose 1996) argues that existing evolutionary models can easily and robustly explain the demographic data. Here we discuss the sensitivity of that analysis to different types of mutational effects, and demonstrate that its conclusion is very sensitive to assumptions about mutations. A legitimate resolution of evolutionary theory and demographic data will require experimental observations on the age-specificity of mutational effects for new mutations and the degree to which mortality rates in adjacent ages are constrained to be similar (positive pleiotropy), as well as consideration of redundancy and heterogeneity models from demographic theory.     

Correlated Changes in Body Melanisation and Mating Traits of Drosophila melanogaster: A Seasonal Analysis

Altitudinal localities of the northern India are associated with high seasonal changes. Drosophila melanogaster flies are darker during the winter season as compared to the autumn season. We tested the hypothesis whether there are altitudinal clines for mating related traits. We observed negative cline for mating latency and positive for copulation period along altitude in D. melanogaster. We further tested if seasonally varying body melanisation is correlated with mating propensity in D. melanogaster. Thus, we examined the D. melanogaster flies collected during autumn and winter season for changes in body melanisation and mating-related traits. Flies from the winter season show high melanisation, copulation duration and fecundity/day as compared to the autumn season flies. By contrast mating latency is longer during autumn as compared to winter season. Based on within- and between-population analysis, body melanisation shows positive correlation with copulation duration and fecundity/day, while negative correlation with mating latency. Within-population analyses show no correlation between body size and ovariole number with body melanisation. Thus, our data suggest that seasonal changes in body melanisation are correlated with mating latency, copulation duration and fecundity/day, but no correlation with body size and ovariole numbers. Further, we observed that seasonal changes in these clines, although have some component of plasticity, have strong genetic basis as the seasonal and population differences were maintained for various traits after 8 generations in the laboratory.     

SPONTANEOUS MUTATIONAL GENOTYPE-ENVIRONMENT INTERACTION FOR FITNESS-RELATED TRAITS IN DROSOPHILA MELANOGASTER

Spontaneous mutations were allowed to accumulate for 104–161 generations in 113–176 inbred lines, independently maintained by a single brother-sister mating per generation, all of them derived from a completely homozygous population of Drosophila melanogaster. In each of two to three consecutive generations, all lines were scored for fecundity, egg-to-pupa and pupa-to-adult viabilities, both in the standard laboratory culture medium (ST) and in three harsh media differing from the former by a single factor: higher temperature (HT), higher NaCl concentration (HSC), or a much reduced concentration of nutrients (D). Relative to the standard medium, productivity (fecundity × viability) decreased by 25% (HT), 66% (HSC), and 80% (D). In each medium, mutational variances of those traits and mutational covariances between all possible pairs were calculated from the between-line divergence (codivergence). Mutational correlations between character states in different media were also obtained. Because we used inbred lines, those estimates were mainly due to the accumulation of mildly detrimental mutations, deleterious mutations of large effect being underrepresented. For all traits, mutational heritabilities ranged from 1.41 × 10^–4 to 11.24 × 10^–4, and did not increase with intensified environmental harshness. Mutational correlations between character states in different media were usually not large (average absolute value 0.31), reflecting a high degree of environmental specificity of the mutations involved. In our results, mutations quasi-neutral in ST conditions and mildly detrimental in more stressful media were not, as a class, important. Mutational correlations between fecundity and egg-to-pupa viability were small and positive in all media. Those involving pupa-to-adult viability were positive in HT, nonsignificant in HSC, and negative in ST and D, showing how the genetic covariance structure of quantitative traits in populations may change in variable environments.     

Mutation accumulation may be a minor force in shaping life history traits

Is senescence the adaptive result of tradeoffs between younger and older ages or the nonadaptive burden of deleterious mutations that act at older ages? To shed new light on this unresolved question we combine adaptive and nonadaptive processes in a single model. Our model uses Penna''s bit-strings to capture different age-specific mutational patterns. Each pattern represents a genotype and for each genotype we find the life history strategy that maximizes fitness. Genotypes compete with each other and are subject to selection and to new mutations over generations until equilibrium in gene-frequencies is reached. The mutation-selection equilibrium provides information about mutational load and the differential effects of mutations on a life history trait - the optimal age at maturity. We find that mutations accumulate only at ages with negligible impact on fitness and that mutation accumulation has very little effect on the optimal age at maturity. These results suggest that life histories are largely determined by adaptive processes. The non-adaptive process of mutation accumulation seems to be unimportant at evolutionarily relevant ages.     

Inbreeding and incompatibility in Trichogramma nr. brassicae: evidence and implications for quality control

Inbreeding effects and incompatibility relationships were examined in strains of the egg parasitoid Trichogramma nr brassicae (Hymenoptera: Trichogrammatidae) from southeastern Australia. Crosses between strains provided weak evidence of incompatibility in a few cases. However sex ratio in crosses within strains tended to be more female-biased than in crosses between strains. Inbreeding was imposed for four generations (F>0.59) of sib mating. The fitness of inbred strains was compared to that of outbred strains generated by crossing the inbred strains. No effects of inbreeding were found for any of the four female traits examined (fecundity, body length, head width and hind tibia length), indicating that T. nr. brassicae is not subjected to inbreeding depression. Inbreeding effects were also not found for male mating success as expected for the haploid sex. There were differences among strains for all traits apart from fecundity, indicating heritable variation. Strain differences for fitness measures were uncorrelated with wasp size. The potential use of inbreeding in the quality control of Trichogramma for mass-release is discussed. Inbreeding may be a useful tool in minimising the effects of laboratory adaptation, thereby extending the useful life of a strain.     

Inbreeding depression and male-mating behavior inDrosophila melanogaster

There have been relatively few studies designed to investigate the effects of inbreeding on behavioral traits. To study this phenomenon, five experimental lines ofDrosophila melanogaster made isogenic for chromosome 2 were evaluated for their male-mating ability and, subsequently, male courtship behavior. All lines showed significant reductions in overall mating ability, and males from all of these lines displayed impaired mating behavior, with two lines displaying particularly aberrant courtship patterns. Line 16 displayed an inability to successfully initiate copulation following successful courtship, while line 17 displayed significant reduction in locomotor activity, resulting in virtually no successful courtship or copulatory activity. The implications of these findings for competitive mating ability in wildDrosophila populations are presented. Further, the importance of mating success as a fitness component in the management of potentially highly inbred populations of endangered species is discussed.     

MUTATION,SELECTION, AND THE MAINTENANCE OF LIFE-HISTORY VARIATION IN A NATURAL POPULATION

In an effort to provide insight into the role of mutation in the maintenance of genetic variance for life-history traits, we accumulated spontaneous mutations in 10 sets of clonal replicates of Daphnia pulex for approximately 30 generations and compared the variance generated by mutation with the standing level of variation in the wild population. Mutations for quantitative traits appear to arise at a fairly high rate in this species, on the order of at least 0.6 per character per generation, but have relatively small heterozygous effects, changing the phenotype by less than 2.5% of the mean. The mean persistence time of a new mutation affecting life-history/body-size traits is approximately 40 generations in the natural population, which requires an average selection coefficient against new mutations of approximately 3% in the heterozygous state. These data are consistent with the idea that the vast majority of standing genetic variance for life-history characters may be largely a consequence of the recurrent introduction of transient cohorts of mutations that are at least conditionally deleterious and raise issues about the meaning of conventional measures of standing levels of variation for fitness-related traits.     

Variation in adult life history and stress resistance across five species of<Emphasis Type="Italic">Drosophila</Emphasis>

Dry weight at eclosion, adult lifespan, lifetime fecundity, lipid and carbohydrate content at eclosion, and starvation and desiccation resistance at eclosion were assayed on a long-term laboratory population ofDrosophila melanogaster, and one recently wild-caught population each of four other species ofDrosophila, two from themelanogaster and two from theimmigrans species group. The relationships among trait means across the five species did not conform to expectations based on correlations among these traits inferred from selection studies onD. melanogaster. In particular, the expected positive relationships between fecundity and size/lipid content, lipid content and starvation resistance, carbohydrate (glycogen) content and desiccation resistance, and the expected negative relationship between lifespan and fecundity were not observed. Most traits were strongly positively correlated between sexes across species, except for fractional lipid content and starvation resistance per microgram lipid. For most traits, there was evidence for significant sexual dimorphism but the degree of dimorphism did not vary across species except in the case of adult lifespan, starvation resistance per microgram lipid, and desiccation resistance per microgram carbohydrate. Overall,D. nasuta nasuta andD. sulfurigaster neonasuta (immigrans group) were heavier at eclosion than themelanogaster group species, and tended to have somewhat higher absolute lipid content and starvation resistance. Yet, these twoimmigrans group species were shorter-lived and had lower average daily fecundity than themelanogaster group species. The smallest species,D. malerkotliana (melanogaster group), had relatively high daily fecundity, intermediate lifespan and high fractional lipid content, especially in females.D. ananassae (melanogaster group) had the highest absolute and fractional carbohydrate content, but its desiccation resistance per microgram carbohydrate was the lowest among the five species. In terms of overall performance, the laboratory population ofD. melanogaster was clearly superior, under laboratory conditions, to the other four species if adult lifespan, lifetime fecundity, average daily fecundity, and absolute starvation and desiccation resistance are considered. This finding is contrary to several recent reports of substantially higher adult lifespan and stress resistance in recently wild-caught flies, relative to flies maintained for a long time in discretegeneration laboratory cultures. Possible explanations for these apparent anomalies are discussed in the context of the differing selection pressures likely to be experienced byDrosophila populations in laboratory versus wild environments. This paper is dedicated to the memory of our friend and former colleague Dr Hans Raj Negi, who tragically passed away at a very young age in a road accident in November 2003.     

ACCLIMATION,CROSS-GENERATION EFFECTS,AND THE RESPONSE TO SELECTION FOR INCREASED COLD RESISTANCE IN DROSOPHILA

The way populations respond to selection can be altered when populations are acclimated prior to selection. To examine this possibility, the responses of replicate lines of Drosophila melanogaster and D. simulans to selection for increased resistance to cold were compared. Flies were selected without hardening or after they had been hardened by holding them at 4°C for one hour. The selection response in both species was much greater when flies were not cold-hardened. Cold resistance in both sets of selected lines reached a plateau after a few generations. Surprisingly, continued selection for increased resistance resulted in decreasing levels of resistance. This decrease was no longer evident after selection had been relaxed for a generation, suggesting cross-generation effects. The magnitude of the cross-generation effects increased with additional generations of selection. Cross-generation effects were also detected for fitness components. Relaxing selection for a generation increased fecundity, weight, viability, and development time. Comparisons of relaxed lines and control lines indicated that only fecundity was influenced by selection. Both sets of selected lines had a lower fecundity than control lines. Crosses between control and selected lines and among replicate selected lines indicated that this decrease in fecundity was not associated with inbreeding. The direct and correlated responses to selection for cold resistance can therefore be influenced by acclimation and cross-generation effects.     

Phenology,fecundity and life table parameters of the predator Hippodamia variegata reared on Dysaphis crataegi

Phenological and some biological characteristics of Hippodamia (Adonia) variegata (Goeze) (Coleoptera: Coccinellidae), such as voltinism, hibernation, number of progeny produced by each generation, mating activity and sex ratio were studied, in order to evaluate the significance of this predator. The study of phenology was conducted in outdoor cages in Kifissia (Athens), during 1999--2001 and as prey was given Dysaphis crataegi (Kaltenbach). Hippodamia variegata completed seven generations between April and November. The hibernating population of H. variegata consisted of adults of 6th and 7th generations. The fecundity of the predator was studied under constant conditions [25 °C, 65% R.H. and 16:8(L:D)h photoperiod] in the laboratory and some population parameters were calculated: The total fecundity ranged between 789 and 1256 eggs, while the mean total fecundity was 959.6 eggs. The greatest proportion of eggs (45%) was oviposited in clutches of 11--20 eggs. The net reproductive value (R₀) was found to be 425.9 females/female, the intrinsic rate of increase (r_m) 0.178 females/female/day and the mean generation time (T) 34.0 days.