Genetic analysis of extended lifespan in Drosophila melanogaster III. On the relationship between artificially selected and wild stocks

Adult lifespans, age‐specific survival, age‐specific mortality, survival times on paraquat, and survival times on DDT were assayed in seven lines of Drosophila melanogaster, including two genetically heterogeneous wild lines recently collected from nature, and three inbred and recombinant inbred lines derived from an artificial selection experiment for increased lifespan. Survival on paraquat is positively correlated with adult lifespan. DDT resistance is uncorrelated with either paraquat resistance or lifespan. The wild lines are unexceptional with respect to average lifespan, paraquat resistance, age‐specific survivorship, and leveling off of mortality rates at advanced ages, but have high levels of DDT resistance. Cluster analysis groups the wild lines with three unselected laboratory stocks in one cluster, while two long‐lived elite recombinant inbred lines form a second cluster. Long‐lived laboratory‐adapted lines are quantitatively differentiated from the wild stocks, both with respect to average adult lifespans and resistance to an oxidizing agent. We reject the ‘recovery’ hypothesis, which proposes that Drosophila artificially selected for long life have phenotypes that merely recover the wild state. This revised version was published online in July 2006 with corrections to the Cover Date.     

Correlated responses to selection for wing length in allozyme systems of Drosophila melanogaster

Summary Significant changes of genotypic structure in 20 lines selected for wing length are detected by analysis of the allelic frequencies of several enzyme loci (XDH, LAP-D, EST-6, 1-APH, ADH, -GPDH). These changes are not haphazard but a consequence of the effects of selection on the genetic structure of the population, since replicate lines always behave in a parallel way. The changes are larger in the lines selected for short wings, in which the genetic variability decreases considerably. This decrease is the result of selection for homozygosity, detected at the allozyme loci, but most probably reflects homozygosity of more or less extended chromosomal segments. Selection for wing length, especially for short wings, favoured recombinants of the initial founder chromosomes. Only in the 1-APH and the EST-6 loci, separated by 11.7 centimorgans on the genetic map, do the alleles linked in the founder lines change in parallel in the control and long wing lines. The correlated response in the allozyme allele frequencies cannot be accounted for by a direct influence of the allozymes on the variability in wing length. The changes in the EST-6, 1-APH and perhaps in the LAP-D, can be explained by a direct effect of natural selection on the allozyme loci, probably in interaction with the effect of selection for wing length on linked loci. This last effect seems to be the main factor contributing to the change detected in the XDH locus.     

Longevity and resistance to cold stress in cold‐stress selected lines and their controls in Drosophila melanogaster

Thermal environments can influence many fitness‐related traits including life span. Here, we assess whether longevity in Drosophila melanogaster can experimentally evolve as a correlated response to cold‐stress selection, and whether genotype‐by‐temperature and sex‐by‐temperature interactions are significant components of variation in life span. Three replicated S lines were cold‐stress selected and compared with their respective unselected controls (Clines) in the 16^th generation of thermal selection. Cold‐stress resistance exhibited a substantial direct response to selection, and also showed a significant interaction between sex and type of line. Mean longevity exhibited a significant interaction between adult test temperature (14 and 25 °C) and line (with suggestive evidence for increased longevity of S lines when tested at 14 °C), but there was no evidence for increased longevity in S lines at normal temperatures (i.e. 25 °C). Another temperature‐dependent effect was sex‐specific, with males being the longer lived sex at 25 °C but the less long‐lived sex at 14 °C. Additionally, we tested in an exploratory way the relationship between longevity and cold‐stress resistance by also measuring resistance to a prefreezing temperature before and after one generation of longevity selection at 14 °C (selection intensity, i = 1.47 for S lines, and 1.42 for C lines). In this longevity selection, we found that cold‐stress resistance increased by about 6% in S lines and 18% in C lines. However, taken together, the results indicate no simple relationship between longevity and cold‐stress resistance, with genotype‐by‐sex interactions in both traits. Temperature dependent interaction in longevity is apparent between S and C lines, and sex‐specific variation in mean longevity also depends on temperature.     

The impact of genetic parental distance on developmental stability and fitness in Drosophila buzzatii

Measures of genetic parental distances (GPD) based on microsatellite loci (D (2) and IR), have been suggested to be better correlated with fitness than individual heterozygosity (H), as they contain information about past events of inbreeding or admixture. We investigated if GPD increased with increasing genetic divergence between parental populations in Drosophila buzzatii and if the measures indicate past events of admixture. Further we evaluated the relationship between GPD, fitness and fluctuating asymmetry (FA) of size and shape. We investigated three populations of Drosophila buzzati, from Argentina, Europe and Australia. From these populations two intraspecific hybridisation lines were made; one between the Argentinean and European populations, which have been separated 200 years and one between the populations from Argentina and Australia, which have been separated 80 years. By doing this we obtained hybrid progeny having different levels of GPD. We found that D (2) and H can be used as indicators of admixture when comparing hybrid individuals with their parentals. IR was not informative. Our results does not exclude the presence of genetic fitness correlations (GFC) over individuals with a broad fitness range from populations in equilibrium, but we doubt the presence of GFC using GPD measures in admixed populations. Shape FA could be a relevant measure for fitness, however, only when comparing populations, not at individual level.     

Allozyme genotype-environment relationships in natural populations of Drosophila buzzatii

Allozyme frequency data from natural populations of Drosophila buzzatii were analyzed for genotype-environment relationships. Allele frequency and heterozygosity at six loci polymorphic throughout eastern Australia and a number of environmental factors (both means and variabilities) were examined by a variety of multivariate techniques. Significant genotype-environment associations were found for five of the six loci, and after correcting for geographic location significant associations remained for Est-2 and Adh-1 gene frequencies and heterozygosities and for Pgm gene frequencies. The results are discussed in relation to selection and gene flow and provide the basis for laboratory studies to disentangle confounded effects of (1) environmental means and environmental variabilities and (2) allele frequency and heterozygosity, and thus to further test for and determine the nature of any natural selection at particular allozyme loci.This work was supported by a grant from the Australian Research Grants Committee to J. S. F. B.     

Visual control of wing beat frequency in Drosophila

The present study shows that the wing beat frequency of Drosophila is visually controlled and modulated in response to different optomotor stimuli.Whereas rotational large field stimuli do not appear to modulate wing beat frequency, single rotating vertical stripes increase or decrease wing beat frequency when moving back-to-front or front-to-back, respectively. Maximal modulations occur at lateral stripe positions.Expansion stimuli eliciting the landing response cause a marked increase in wing beat frequency. Parameters of this frequency response depend in a graded fashion on certain stimulus properties, and the frequency response co-habituates with the landing response. Several results indicate that the frequency response is an integral component of the landing response, although it can also occur when the characteristic front leg extension is not observed. The complex spatial input integration underlying the frequency response and other motor components of the landing response cannot easily be explained by a system of large field integration units, but might indicate the existence of local expansion detectors.     

Differential mutagenic response in selected lines of Drosophila melanogaster

Summary The mutagenic efficiency of ionizing radiations has been tested on different lines of Drosophila melanogaster. It has been shown that differential lethal effects are obtained when irradiated females from different lines are mated to flies carrying heterozygous lethal genes. The results seem not to be attributable to differential expression of the lethality in the various crosses performed with the irradiated flies. This might suggest that gene activity is involved in the expression of the mutagenic effects of radiations.     

Spatial differences in patterns of modification: selection on hairy in Drosophila melanogaster wings

Artificial selection was carried out for over 45 generations to enhance and suppress expression of the mutation hairy on the Drosophila melanogaster wing. Whole chromosome mapping of X‐linked and autosomal modifiers of sense organ number displayed regional differences in magnitude and direction of their effects. Regional specificity of modifier effects was also seen in some interchromosomal interactions. Scanning electron microscopy allowed precise measurement of sense organ size and position along the L3 longitudinal wing vein. Sense organ size varied in a predictable fashion along the proximal–distal axis, and the dorsal pattern differed from the ventral pattern. The high and low selection lines differed most in the proximal portion of the L3 vein. Extra sense organs in the High line were often associated with vein fragments at locations predicted from ancestral vein patterns. Thus, regional specificity of polygenic or quantitative trait locus modifier effects was identified in several different parts of the wing. This revised version was published online in July 2006 with corrections to the Cover Date.     

Heterosis in crosses between lines of Drosophila melanogaster selected for adaptation to different environments

Summary Experiments were designed to examine whether heterosis would occur in crosses of Drosophila melanogaster populations adapted to 18 °C or 28 °C environments. Crosses were examined in parental environments, an intermediate environment (23 °C) and a mixed environment (alternating 18°/28°C). Parental populations did not show divergence for larval viability, cold shock or high temperature mortalities when tested in a common environment. However, the 28 °C population was less fecund than the 18 °C population, but had higher larval competitive ability and higher adult longevity. Heterosis for viability, cold shock mortality and high temperature mortality occurred in crosses between a population adapted to 18 °C and another adapted to 28 °C, but not in crosses between two populations adapted to the same temperature. The results suggest that, in the absence of drift, heterosis is expected in crosses between lines or populations with different histories of selection but not between lines with the same selection histories.     

Genomic distribution of transposable elements among individuals of an inbred Drosophila line

The stability of the elements of eleven transposon families (412, B 104, blood, 297, 1731, G, copia, mdg 4, hobo, jockey and I) has been compared by the Southern technique among individuals of a Drosophila line that has been subjected to 30 generations of sister sib matings. The 412, B104, blood, 297, 1731 and G elements appear stable. Heterochromatic copia and hobo elements and euchromatic I elements appear highly polymorphic. In addition, copia, mdg 4, jockey and I elements undergo an instability resulting in significant variations in relative intensity among autoradiographic bands. The extent of the polymorphisms detected strongly suggests de novo rearrangements of transposable elements.     

A study of wing morphology and fluctuating asymmetry in interspecific hybrids between Drosophila buzzatii and D. koepferae

In this work we investigate the effect of interspecific hybridization on wing morphology using geometric morphometrics in the cactophilic sibling species D. buzzatii and D. koepferae. Wing morphology in F₁ hybrids exhibited an important degree of phenotypic plasticity and differs significantly from both parental species. However, the pattern of morphological variation between hybrids and the parental strains varied between wing size and wing shape, across rearing media, sexes, and crosses, suggesting a complex genetic architecture underlying divergence in wing morphology. Even though there was significant fluctuating asymmetry for both, wing size and shape in F₁ hybrids and both parental species, there was no evidence of an increased degree of fluctuating asymmetry in hybrids as compared to parental species. These results are interpreted in terms of developmental stability as a function of a balance between levels of heterozygosity and the disruption of coadaptation as an indirect consequence of genomic divergence.     

Altitudinal patterns for longevity, fecundity and senescence in Drosophila buzzatii

We tested for variation in longevity, senescence rate and early fecundity of Drosophila buzzatii along an elevational transect in Argentina, using laboratory-reared flies in laboratory tests performed to avoid extrinsic mortality. At 25 °C, females from lowland populations lived longer and had a lower demographic rate of senescence than females from highland populations. Minimal instead of maximal temperature at the sites of origin of population best predicted this cline. A very different pattern was found at higher test temperature. At 29.5 °C, longevity of males increased with altitude of origin of population. No clinal trend was apparent for longevity of females at 29.5 °C. There was evidence for a trade-off between early fecundity and longevity at non-stressful temperature (25 °C) along the altitudinal gradient. This trait association is consistent with evolutionary theories of aging. Population-by-temperature and sex-by-temperature interactions indicate that senescence patterns are expressed in environment specific ways.     

Embryonic cAMP and developmental potential in Drosophila melanogaster

Summary Measurements of cAMP in early embryos of Drosophila melanogaster demonstrate that the dunce gene plays a major role, and the rutabaga gene a secondary role, in maternal regulation of embryonic cAMP content. Studying the double mutant combination, we find that variability in elevated cAMP content between individual embryos is associated with a wide variability in developmental potential. Embryos with about five times the normal cAMP content define a threshold between apparently normal and abnormal development. Measurements of cAMP content in anterior and posterior halves of embryos indicate that the posterior embryonic region, which is developmentally more sensitive to the effects of elevated cAMP than the anterior region, does not contain more cAMP than the anterior region. The variety of developmental defects observed is discussed in relation to possible targets of cAMP action.Offprint requests to: J.A. Kiger, Jr     

Size-related sexual selection and yeast diet inDrosophila buzzatii (Diptera: Drosophilidae)

Size-related sexual selection (SRSS) was examined on four traits (thorax and wing length and head and face width) inDrosophila buzzatii, by scoring male copulatory status in two mass-mating experiments. Using axenic females, experiment 1 was carried out with axenic males, and experiment 2 with yeast-supplemented males. While there was no indication of SRSS in experiment 1, such selection was substantial in yeast-supplemented males, which transmitted yeasts to mating females. Multivariate analyses of selection indicated that face width is the measured trait on which directional SRSS essentially acted in yeast-supplemented males, resulting in indirect selection on body size. Because this selection was affected by yeast diet in males, its possible interaction with the yeast transmission from males to females during the courtship is discussed.     

Genetic analysis of extended life span in Drosophila melanogaster I. RAPD screen for genetic divergence between selected and control lines

Using lines selected for long life by Luckinbill and his co-workers, we screened two selected and two control lines for allelic frequency differences at 1200 randomly chosen RAPD marker loci. Twenty-three marker loci showed frequency differences in excess of 80%, and five were greater than 90%. Age-specific effects of the five most differentiated loci were estimated by collecting complete survival data in segregating backcross populations. Alleles at four of the five marker loci were associated with significant extension of life span in males, while two marker loci had significant effects in females. Eighty percent of the total selection response in males can be explained by the identified QTL's, under the assumption of additivity. The N14+ marker allele accounted for a 12-day life span extension in males, but had little effect in females. Both sex-limited and sex-shared effects were observed. Analysis of age-specific mortality rates suggests that life span extension occurs by a combination of genetic factors that moderate both the level of mortality and the rate at which mortality increases with age. This revised version was published online in July 2006 with corrections to the Cover Date.     

Second-site modifiers of the Delta wing phenotype in Drosophila melanogaster

Summary We have screened for dominant enhancers and suppressors of the wing phenotype associated with two Delta alleles: Dl ^9P39, an amorphic allele, and Dl ^FE32, an antimorphic allele. The interactions of some of the modifiers with Delta are due to haplo-insufficient expression of the corresponding genes. Although not explicitly shown for the remaining cases, we assume that haploin-sufficiency is also the basis for the relationships of these genes to Delta, since no allele specific interactions were observed. The modifiers found define 22 genes with pleiotropic expression, which can be classified into two groups: genes required for wing vein pattern formation and for neurogenesis, and genes which are not required for neurogenesis. Among the genes of the first group, Hairless and Star were previously known to participate in neural development. One further modifier was found which may correspond to a new neurogenic gene. The second group of genes is larger and includes already known loci, e.g., Plexate, blistered, plexus, etc, as well as other previously unidentified genes, which function during wing morphogenesis. Correspondence to: J.A. Campos-Ortega     

The cell biology of Drosophila wing metamorphosis in vitro

Summary We have examined the metamorphosis of the wing imaginal disc of Drosophila during culture in vitro in the continuous presence of 20-hydroxy ecdysone (0.1 g/ ml). We find that the sequence of cellular changes in the wing blade during culture closely match those occurring in situ, involving two periods at which the dorsal and ventral surfaces are joined only by cell processes containing trans-alar microtubule arrays. Good pupal and imaginal cuticle secretion is found in this system.     

Effect of Temperature on Longevity and Productivity in Drosophila Ananassae: Evidence for Adaptive Plasticity and Trade-Off Between Longevity and Productivity

The genetic response of body size to temperature in the laboratory provides an interesting example of phenotypic plasticity. We found that females of Drosophila ananassae reared to adulthood at 18°C showed significant increase in body weight as compared to females reared at 25°C. At a given temperature, early productivity and lifetime productivity were the highest when the rearing and test temperature were the same. The effect of test temperature was highly significant for total productivity and early productivity. The interaction between test temperature and development temperature was also highly significant. Effect of development temperature was not significant. The females reared at 18°C showed greater body weight but their productivity was not significantly higher than smaller females reared at 25°C. Thus, the usually close relationship between size and fecundity is lost when the size change is due to rearing temperature. These findings provide evidence for adaptive plasticity in D. ananassae. We also found a negative correlation (trade-off) between longevity and productivity, the first report of such a trade-off between longevity and productivity in D. ananassae.     

A three-locus system of interspecific incompatibility underlies male inviability in hybrids between Drosophila buzzatii and D. koepferae

In hybrids between the sibling species D. buzzatii and D. koepferae, both sexes are more or less equally viable in the F₁: However, backcross males to D. buzzatii are frequently inviable, apparently because of interspecific genetic incompatibilities that are cryptic in the F₁. We have performed a genetic dissection of the effects of the X chromosome from D. koepferae. We found only two cytological regions, termed hmi-1 and hmi-2, altogether representing 9% of the whole chromosome, which when introgressed into D. buzzatii cause inviability of hybrid males. Observation of the pattern of asynapsis of polytene chromosomes (incomplete pairing, marking introgressed material) in females and segregation analyses were the technique used to infer the X chromosome regions responsible for this hybrid male inviability. The comparison of these results with those previously obtained with the same technique for hybrid male sterility in this same species pair indicate that in the X chromosome of D. koepferae there are at least seven times more regions that produce hybrid male sterility than hybrid male inviability. We have also found that the inviability brought about by the introgression of hmi-1 is suppressed by the cointrogression of two autosomal sections from D. koepferae. Apparently, these three regions conform to a system of species-specific complementary factors involved in an X-autosome interaction that, when disrupted in backcross hybrids by recombination with the genome of its sibling D. buzzatii, brings about hybrid male inviability.     

Accumulation of lethals in highly selected lines of Drosophila melanogaster

Summary Four synthetic lines of D. melanogaster selected for low sternopleural bristle number for 50 generations were screened for lethals on chromosome III when their mean score equalled 2.5. Each line originated from a cross between line M (previously selected for the same trait during 130 generations) and a different unselected cage population. Line M was already known to carry a recessive lethal on chromosome III affecting the selected trait, such that the bristle score of the lethal heterozygote was lower than that of the viable homozygote. Tests revealed 18 lethals, 15 of these present in at least two lines. Each line carried from 10 to 16 lethals. All lines carried groups of lethals present on the same chromosome, and at least six lethals in each line were included in such an association with a frequency of 0.18 or higher. It appears that the lethal affecting bristle score in line M has protected a segment of chromosome III from natural selection and that the remaining 14 lethals have accumulated later in that line.