Condition dependence and the nature of genetic variation for male sex comb bristle number in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

Genetic architecture of variation underlying male sex comb bristle number, a rapidly evolving secondary sexual character of Drosophila, was examined. First, in order to test for condition dependence, diet was manipulated in a set of ten Drosophila melanogaster full-sib families. We confirmed heightened condition dependent expression of sex comb bristle number and its female homologue (distal transverse row bristles) as compared to non-sex sternopleural bristles. Significant genotype by environment effects were detected for the sex traits indicating a genetic basis for condition dependence. Next we measured sex comb bristle number and sternopleural bristle number, as well as residual mass, a commonly used condition index, in a set of thirty half-sib families. Sire effect was not significant for sex comb and sternopleural bristle number, and we detected a strong dominance and/or maternal effect or X chromosome effect for both traits. A strong sire effect was detected for condition and its heritability was the highest as compared to sex comb and sternopleural bristles. We discuss our results in light of the rapid response to divergent artificial selection for sex comb bristle number reported previously. The nature of genetic variation for male sex traits continues to be an important unresolved issue in evolutionary biology.     

Developmental stress in wild-living Drosophilids inferred from biometry: metric and meristic traits react differently to heterogeneous environmental conditions

Abstract.  1. During their development in natural conditions, Drosophila larvae and pupae face heterogeneous environmental conditions (HEC). Analysing the morphology of wild-living adults is a means of evaluating the effects of HEC.
2. Two drosophilid species of the Zaprionus genus that recently invaded the Nile delta were investigated, and three metric, size-related traits, and one meristic trait, the sternopleural bristle number, were measured. Data were compared with those of F₁ generation reared under favourable laboratory conditions.
3. Body size was smaller in nature, but also extremely variable among individuals, with an average coefficient of variation (CV) of 9.1 ± 0.3, against a much lesser value of 2.4 ± 0.1 in laboratory flies. Correlations among size traits were also greater in nature (0.96 ± 0.01) than in the laboratory (0.75 ± 0.04).
4. By contrast, sternopleural bristles produced similar results in wild-living and laboratory flies. In nature, mean numbers were only slightly less than in the laboratory and the CVs were almost identical (10.87 ± 0.70 vs 10.80 ± 0.76).
5. Fluctuating asymmetry of sternopleural bristles was also identical in the two kinds of flies, and was not influenced by HEC, indicating a high level of developmental canalisation with respect to HEC.
6. The implications of the results for the problems of field heritability and developmental stability are discussed.     

Genetic Variability and Fluctuating Asymmetry of Morphological Traits in Drosophila melanogasterReared on a Pesticide-Containing Medium

The effects of chlorine-organic insecticide endosulfane (thiodan) on phenotypic and genetic variation in four morphological traits of Drosophila melanogaster(wing length, thorax length, the number of orbital bristles and the number of sternopleural bristles) were examined. In addition, the effect of this pesticide on stability of development measured as fluctuating asymmetry of bilateral traits was estimated. On the medium with endosulfane, phenotypic variation of morphometric traits was significantly higher. No difference in fluctuating asymmetry between the stressed and the control samples was found. The among-line variance of morphometric traits of flies reared on the endosulfane-containing medium was significantly higher as compared to the corresponding variance under control conditions. The efficiency of using fluctuating asymmetry and phenotypic variation of morphometric and meristic traits as indicators of environmental stress in insect populations is discussed.     

Arrangement of bristles as a function of bristle number on a leg segment inDrosophila melanogaster

Summary The arrangement of bristles on a leg segment of the fruitflyDrosophila melanogaster was studied in various mutants that have abnormal numbers of bristles on this segment. Eighteen mutations at six different genetic loci were analyzed, plus five double or triple mutant combinations. Recessive mutations at theachaete-scute locus were found to affect distinct groups of bristles:achaete mutations remove mechanosensory bristles, whereasscute mutations remove mainly chemosensory bristles. Mechanosensory bristles remain uniformly spaced along the longitudinal axis unless their number decreases below a certain threshold, suggesting that spacing is controlled by cell interactions that cannot function when bristle cells are too far apart. Above a certain threshold, bristle spacing and alignment both become irregular, perhaps due to excessive force from these same interactions. Chemosensory bristles occupy definite positions that are virtually unaffected by removal of individual bristles from the array. Extra chemosensory bristles develop only near the six normal sites. At two of the six sites the multiple bristles tend to exhibit uniform longitudinal spacing — a property confined to mechanosensory bristles in wild-type flies. To explain the various mutant phenotypes the following scheme is proposed, with different mutations directly or indirectly affecting each step: (1) spots and stripes are demarcated within the pattern area, (2) one bristle cell normally arises within each spot, multiple bristle cells within each stripe, (3) incipient bristle cells inhibit neighboring cells from becoming bristle cells, and (4) the bristle cells within each stripe become aligned to form rows and then repel one another to generate uniform spacing.     

Quantitative trait loci responsible for variation in sexually dimorphic traits in Drosophila melanogaster

To understand the mechanisms of morphological evolution and species divergence, it is essential to elucidate the genetic basis of variation in natural populations. Sexually dimorphic characters, which evolve rapidly both within and among species, present attractive models for addressing these questions. In this report, we map quantitative trait loci (QTL) responsible for variation in sexually dimorphic traits (abdominal pigmentation and the number of ventral abdominal bristles and sex comb teeth) in a natural population of Drosophila melanogaster. To capture the pattern of genetic variation present in the wild, a panel of recombinant inbred lines was created from two heterozygous flies taken directly from nature. High-resolution mapping was made possible by cytological markers at the average density of one per 2 cM. We have used a new Bayesian algorithm that allows QTL mapping based on all markers simultaneously. With this approach, we were able to detect small-effect QTL that were not evident in single-marker analyses. Our results show that at least for some sexually dimorphic traits, a small number of QTL account for the majority of genetic variation. The three strongest QTL account for >60% of variation in the number of ventral abdominal bristles. Strikingly, a single QTL accounts for almost 60% of variation in female abdominal pigmentation. This QTL maps to the chromosomal region that Robertson et al. have found to affect female abdominal pigmentation in other populations of D. melanogaster. Using quantitative complementation tests, we demonstrate that this QTL is allelic to the bric a brac gene, whose expression has previously been shown to correlate with interspecific differences in pigmentation. Multiple bab alleles that confer distinct phenotypes appear to segregate in natural populations at appreciable frequencies, suggesting that intraspecific and interspecific variation in abdominal pigmentation may share a similar genetic basis.     

The development and evolution of bristle patterns in Diptera

The spatial distribution of sensory bristles on the notum of different species of Diptera is compared. Species displaying ancestral features have a simple organization of randomly distributed, but uniformly spaced, bristles, whereas species thought to be more derived bear patterns in which the bristles are aligned into longitudinal rows. The number of rows of large bristles on the scutum was probably restricted to four early on in the evolution of cyclorraphous Brachyceran flies. Most species have stereotyped patterns based on modifications of these four rows. The possible constraints placed upon the patterning mechanisms due to growth and moulting within the Diptera are discussed, as well as within hemimetabolous insects. The holometabolic life cycle and the setting aside of groups of imaginal cells whose function is not required during the growth period, may have provided the freedom necessary for the evolution of elaborate bristle patterns. We briefly review the current state of knowledge concerning the complex genetic pathways regulating achaete-scute gene expression and bristle pattern in Drosophila melanogaster, and consider mechanisms for the genetic regulation of the bristle patterns of other species of Diptera.     

Sexual selection for symmetrical male medflies (Diptera: Tephritidae) confirmed in the field

The role of fluctuating asymmetry as an indicator of fitness to females in mate choice remains controversial. Previous studies indicated that male medflies with symmetrical supra-fronto-orbital (SFO) bristles achieve relatively high mating success under laboratory conditions. Here we present data from field cage studies of wild collected medflies in Guatemala and Crete, which reveal the same association between fluctuating asymmetry in SFO bristle length and mating success as that seen in the laboratory. The experiments in Crete included males that were missing one or both of their bristles. A comparison of mating success between the three groups indicated that the mere presence of bristles did not exert a major influence. Analysis of attempted courtships suggests that the association between male mating success and FA in bristle length appears to be generated as a result of females being more likely to enter into courtships with symmetrical males, rather than through a rejection of asymmetrical males during or after it. This raises the possibility that the primary stimulus that makes a symmetrical male attractive is acting at too great a distance to depend on symmetry itself. Alternatives might include superior pheromone emissions or the occupation of a prime location within the lek.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society, 2004, 81 , 347–355.     

Action of natural selection in lines of Drosophila selected for a new level of canalization

Summary Four lines of Drosophila melanogaster previously selected for a stabilized phenotype of two extra dorsocentral bristles were examined for 20 generations of canalizing selection and relaxation of selection. A substantial frequency of flies with either two anterior or two posterior extra bristles was maintained in the relaxed lines. These patterns were the only ones tolerated by natural selection, i.e., the only symmetric ones. It was concluded that anterior and posterior dorsocentral bristles are two independent development structures, and the results are discussed in relation to two proposed genetic systems for bristle determination.     

Genetic variability and fluctuating asymmetry of morphological signs of Drosophila melanogaster during development in a pesticide-containing environment

The effects of chlorine-organic insecticide endosulfane (thiodan) on phenotypic and genetic variation in four morphological traits of Drosophila melanogaster (wing length, thorax length, the number of orbital bristles and the number of sternopleural bristles) were examined. In addition, the effect of this pesticide on stability of development measured as fluctuating asymmetry of bilateral traits was estimated. On the medium with endosulfane, phenotypic variation of morphometric traits was significantly higher. No difference in fluctuating asymmetry between the stressed and the control samples was found. The among-line variance of morphometric traits of flies reared on the endosulfane-containing medium was significantly higher as compared to the corresponding variance under control conditions. The efficiency of using fluctuating asymmetry and phenotypic variation of morphometric and meristic traits as indicators of environmental stress in insect populations is discussed.     

塔里木沙拐枣果实刺毛对果实扩散及萌发的影响

塔里木沙拐枣（Calligonum roborowskii A.Los.）是仅分布于我国西北地区荒漠环境的特有种,其果实为不开裂而周围有椭圆形刺毛的瘦果,以果实为单元进行扩散和萌发。本文采用野外观察与室内控制实验相结合的方法,对塔里木沙拐枣刺毛在果实扩散、吸水、脱水及萌发中的作用进行研究。结果显示：不同贮藏时间的果实颜色、大小、重量及刺毛长度存在显著差异。完整果实在水面漂浮时间及水媒扩散能力比去除刺毛的果实更强。完整果实在1 m/s和4 m/s风速下的扩散距离比无刺毛果实的扩散距离长。10 mmol/L赤霉素（AG₃）及不同干藏时间处理的完整果实与无刺毛果实在20℃/30℃光照和黑暗条件下的萌发率间存在显著差异,其中无刺毛果实的萌发率要高于完整果实的萌发率。室外盆栽实验显示,无刺毛果实的萌发率比完整果实高,说明果实刺毛对果实萌发有抑制作用。研究结果表明果实刺毛对塔里木沙拐枣在塔克拉玛干沙漠边缘荒漠环境中的扩散、萌发及种群更新具有重要的辅助作用。     

A Genetic Analysis of Pannier, a Gene Necessary for Viability of Dorsal Tissues and Bristle Positioning in Drosophila

A genetic and phenotypic analysis of the gene pannier is described. Animals mutant for strong alleles die as embryos in which the cells of the amnioserosa are prematurely lost. This leads to a dorsal cuticular hole. The dorsal-most cells of the imagos are also affected: viable mutants exhibit a cleft along the dorsal midline. pannier mRNA accumulates specifically in the dorsal-most regions of the embryo and the imaginal discs. Viable mutants and mutant combinations also affect the thoracic and head bristle patterns in a complex fashion. Only those bristles within the area of expression of pannier are affected. A large number of alleles have been studied and reveal that pannier may have opposing effects on the expression of achaete and scute leading to a loss or a gain of bristles.     

Condition dependence of developmental stability in the sexually dimorphic fly Telostylinus angusticollis (Diptera: Neriidae)

Developmental stability is widely regarded as a condition‐dependent trait, but its relation to genotype and environment, and extent of developmental integration, remain contentious. In Telostylinus angusticollis, the dorsocentral bristles exhibit striking variation in developmental stability, manifested as fluctuating asymmetry (FA) in bristle position (‘positional FA’) and failure to develop some bristles (‘bristle loss’), in natural and laboratory populations. To determine whether this variation reflects condition, I tested for effects of genotype and environment (larval diet quality), and examined covariation with condition‐dependent traits. Positional FA was not affected by genotype or environment. However, positional FA covaried negatively with secondary sexual trait expression in males, and with sexual dimorphism in body shape, but covaried positively with body size in females. Bristle loss reflected both genotype and larval diet. Flies reared on poor‐quality diet exhibited a similar rate of bristle loss as wild flies. Both positional FA and bristle loss were greater in males. These results suggest that the relation between developmental stability and condition is complex and sex dependent.     

Premolis semirufa (Walker, 1856) envenomation, disease affecting rubber tappers of the Amazon: searching for caterpillar-bristles toxic components

Background

The caterpillar of the moth Premolis semirufa (Lepidoptera: Arctiidae), commonly named Pararama, is endemic of the Amazon basin. Accidental contact with these caterpillar bristles causes local symptoms such as intense heat, pain, edema and itching which last for three to seven days; however, after multiples contacts, it may induce joint-space narrowing and bone alteration, as well as degeneration of the articular cartilage and immobilization of the affected joints. Specific treatment for this disease does not exist, but corticosteroids are frequently administered. Despite of the public health hazard of Premolis semirufa caterpillar poisoning, little is known about the nature of the toxic components involved in the induction of the pathology.

Methodology/Principal Findings

Here we have investigated the biological and immunochemical characteristics of the caterpillar''s bristles components. Analysis of the bristles extract in in vitro assays revealed the presence of proteolytic and hyaluronidase activities but no phospholipase A₂ activity. In vivo, it was observed that the bristles extract is not lethal but can induce an intense inflammatory process, characterized by the presence of neutrophils in the paw tissues of injected mice. Furthermore, the bristles components stimulated an intense and specific antibody response but autoantibodies such as anti-DNA or anti-collagen type II were not detected.

Conclusion

The results suggest that Premolis semirufa caterpillar bristles secretion contains a mixture of different enzymes that may act together in the generation and development of the clinical manifestations of the Pararama envenomation. Moreover, the high immunogenicity of the caterpillar bristles components, as shown by the generation of high antibody titers, may also contribute to the induction and establishment of the inflammatory disease.     

The quantitative genetics of fluctuating asymmetry

Fluctuating asymmetry (subtle departures from identical expression of a trait across an axis of symmetry) in many taxa is under stabilizing selection for reduced asymmetry. However, lack of reliable estimates of genetic parameters for asymmetry variation hampers our ability to predict the evolutionary outcome of this selection. Here we report on a study, based on analysis of variation within and between isofemale lines and of generation means (line-cross analysis), designed to dissect in detail the quantitative genetics of positional fluctuating asymmetry (PFA) in bristle number in natural populations of Drosophila falleni. PFA is defined as the difference between the two sides of the body in the placement or position of components of a meristic trait. Heritability (measured at 25 degrees C) of two related measures of PFA were 13% and 21%, both of which differed significantly from zero. In contrast, heritability estimates for fluctuating asymmetry in the total number of anterior (0.7%) and transverse (2.4%) sternopleural bristles were smaller, not significant, and in quantitative agreement with previously published estimates. Heritabilities for bristle number (trait size) were considerably greater than that for any asymmetry measure. The experimental design controlled for the potentially confounding effects of common familial environment, and repeated testing revealed that PFA differences between lines were genetically stable for up to 16 generations in the laboratory at 25 degrees C. We performed line cross analysis between strains at the extremes of the PFA distribution (highest and lowest values); parental strains, F1, F1r (reciprocal), F2, backcross, and backcross reciprocal generations were represented. The inheritance of PFA was described best by additive and dominance effects localized to the X-chromosomes, whereas autosomal dominance effects were also detected. Epistatic, maternal, and cytoplasmic effects were not detected. The inheritance of trait size was notably more complex and involved significant autosomal additive, dominance, and epistatic effects; maternal dominance effects; and additive and dominance effects localized to the X-chromosomes. The additive genetic correlation between PFA and its associated measure of trait size was negative (-0.049), but not statistically significant, indicating that the loci contributing additive genetic effects to these traits are probably different. It is suggested that PFA may be a sensitive measure of developmental instability because PFA taps the ability of an organism to integrate interconnected developmental pathways.     

Polygenic Mutation in Drosophila Melanogaster: Genetic Analysis of Selection Lines

We have conducted genetic analyses of 12 long-term selection lines of Drosophila melanogaster derived from a highly inbred base population, containing new mutations affecting abdominal and sternopleural bristle number. Biometric analysis of the number of effective factors differentiating the selected lines from the base inbred indicated that with the exception of the three lines selected for increased number of abdominal bristles, three or more mutations contributed to the responses of the selection lines. Analysis of the chromosomal distribution of effects revealed that mutations affecting abdominal bristle number occurred on all three major chromosomes. In addition, Y-linked mutations with effects ranging from one to three bristles occurred in all three lines selected for decreased number of abdominal bristles, as well as in one line selected for increased abdominal bristle number. Mutations affecting sternopleural bristle number were mainly on the X and third chromosomes. One abdominal and one sternopleural selection line showed evidence of a segregating lethal with large effects on bristle number. As an indirect test for allelism of mutations occurring in different selection lines, the three lines selected in the same direction for the same trait were crossed in all possible combinations, and selection continued from the F(2) hybrids. Responses of the hybrid lines usually did not exceed those of the most extreme parental lines, indicating that the responses of the parental lines may have been partly due to mutations at the same loci, although other interpretations are possible.     

Variability of morphological traits in Drosophila bipectinata complex

Phenotypic or morphological differences among different populations and sexual dimorphism in certain metric traits were analysed in D. bipectinata complex. It was noticed that different populations of D. bipectinata species group harbour large amount of variation for these characters. In all the populations, morphometric characters such as lengths of femur, tibia and wing length, wing width, number of sternopleural bristles and bristles on epandrium varied significantly among populations. The study indicates that the morphological variations are due to the interplay of genetic and environmental endowments. Further, females had significantly larger values, for lengths of femur, tibia and wing length, wing width and sternopleural bristles.     

Interspecies variability of number of bristles on dorsal surface of aedeagus in D. virilis species group and its genetic mapping with interspecies hybrids of D. virilis and D. lummei

The results of morphologic and hybrid analyses of the feature of the male reproductive system of sibling species in the virilis group were presented. Bristles appeared on the surfaces of male genitals (aedeagus). The occurrence of a specific expression of the examined feature in the phyllades of D. virilis group, the correspondence of both the number and distribution pattern of the bristles on surfaces of the aedeagus and developmental temperature in D. virilis and D. lummei, as well as the link between feature and sexual behavior, have been shown. Dominance of D. lummei phenotype in the interspecies D. virilis × D. lummei was found. The interspecies hybrids D. virilis and D. lummei were used for a genetic analysis of the variability of the examined feature. The significant influence of chromosomes 2 and 6 on the number of bristles on the aedeagus in hybrid males was shown. Furthermore, the correspondence between the effects of the autosomes 2 and 6 on the variability of the examined feature and the genetic status of the other chromosomes (the effect of interaction between genetic factors, chromosomes here) was revealed. The adaptive value of the examined feature related to the involvement in the formation of isolating barriers at the copulation stage is under discussion.     

Systematic Mapping of Protein Mutational Space by Prolonged Drift Reveals the Deleterious Effects of Seemingly Neutral Mutations

Systematic mappings of the effects of protein mutations are becoming increasingly popular. Unexpectedly, these experiments often find that proteins are tolerant to most amino acid substitutions, including substitutions in positions that are highly conserved in nature. To obtain a more realistic distribution of the effects of protein mutations, we applied a laboratory drift comprising 17 rounds of random mutagenesis and selection of M.HaeIII, a DNA methyltransferase. During this drift, multiple mutations gradually accumulated. Deep sequencing of the drifted gene ensembles allowed determination of the relative effects of all possible single nucleotide mutations. Despite being averaged across many different genetic backgrounds, about 67% of all nonsynonymous, missense mutations were evidently deleterious, and an additional 16% were likely to be deleterious. In the early generations, the frequency of most deleterious mutations remained high. However, by the 17th generation, their frequency was consistently reduced, and those remaining were accepted alongside compensatory mutations. The tolerance to mutations measured in this laboratory drift correlated with sequence exchanges seen in M.HaeIII’s natural orthologs. The biophysical constraints dictating purging in nature and in this laboratory drift also seemed to overlap. Our experiment therefore provides an improved method for measuring the effects of protein mutations that more closely replicates the natural evolutionary forces, and thereby a more realistic view of the mutational space of proteins.     

SECRETION AND DEPLOYMENT OF BRISTLES IN MALLOMONAS SPLENDENS (SYNUROPHYCEAE)1

Mallomonas splendens (G. S. West) Playfair has a cell covering of siliceous scales and bristles. Interphase cells bear four anterior and four posterior bristles that each articulate, at their flexed basal ends via a complex of labile fibers (the fibrillar complex), on a specialized body scale (a base-plate scale). Body scales, base-plate scales and bristles are formed independently of each other and at different times in silica deposition vesicles (SDVs) that are associated with one of the two chloroplasts. The fine structure of scale and bristle morphogenesis in M. splendens agrees with that previously described for Synura and Mallomonas. Four new posterior bristles are formed at late interphase with their basal ends towards the cell posterior. The fibrillar complex is formed in situ on the bristle in the SDV. Mature bristles are secreted one by one onto the surface of the protoplast, beneath the layer of body scales, where the basal ends of the bristles adhere to the plasma membrane via the fibrillar complex. The extrusion of posterior bristles and their deployment onto the cell surface was monitored with video. A fine cellular protuberance accompanies the bristles as they are extruded from beneath the scale layer with their basal ends leading. When distant from the cell, the basal ends of the bristles appear attached to the protuberance, possibly by way of their fibrillar complexes. Once bristles are fully extruded, and their tips free in the surrounding environment, the bristle bases are drawn back to the posterior apex of the cell, apparently by the now shortening protuberance. Thus a 180° reorientation of the posterior bristles has been effected outside the cell. Thin-sections of cells that are extruding bristles show a threadlike, cytoplasmic extension of the cell posterior which may be analogous to the protuberance seen in live cells. Four new posterior base-plate scales are secreted after the bristles have reoriented. Scanning electron microscopy indicates that the fibrillar complex is involved in positioning the bristles onto their respective base-plate scales. Anterior bristles are formed in new daughter cells in the same orientation as the posterior bristles; thus they are extruded tip first and no reorientation is required.     

Phenotypic plasticity and reaction norms of abdominal bristle number inDrosophila melanogaster

The phenotypic plasticity of abdominal bristle number (segments 3 and 4 in females) was investigated in 10 isofemale lines from a French population, grown at 7 constant temperatures, ranging from 12‡ to 31‡C. Overall concave reaction norms were obtained with a maximum around 20‡-21‡C. Intraclass correlation (isofemale line heritability) was not affected by temperature. Correlations between segments 3 and 4 strongly contrasted a low within-line phenotypic correlation (r = 0.39 ± 0.04) and a high, between-line genetic correlation (r = 0.89 ± 0.03). A significant decrease of the genetic correlation was observed when comparing more different temperatures. Finally, among 7 other morphometrical traits which were measured on the same set of lines, 3 provided a significant positive genetic correlation with abdominal bristles: thoracic bristles, abdomen pigmentation and thoracic pigmentation.