Abdominal pigmentation and growth temperature in Indian Drosophila melanogaster: Evidence for genotype-environment interaction

Phenotypic variability for abdominal pigmentation in females of an Indian natural population ofDrosophila melanogaster was studied using isofemale lines and by rearing the larvae and pupae at 4 different temperatures ranging from 20–30°C. The dark pigmented area was found to increase in all the three segments when the growth temperature decreases. A significant positive correlation was detected for the occurrence of dark pigmentation in the 5th and 6th segments in each growth temperature but for other comparisons the correlation was not regular. Analysis of variance (ANOVA) was carried out both for individual segments over different growth temperatures and also for each temperature over the three abdominal segments and in all cases found to be statistically significant. The results are quite different from the earlier observation in FrenchDrosophila melanogaster and suggest that genes controlling pigmentation are temperature dependent; temperature could affect post-transitional events involved in pigmentation. The present findings also clearly indicate that significant genotype-environment interaction exists, responsible for the production of desired phenotype at the opportune moment during the life span of a species.     

THE INFLUENCE OF ABDOMINAL PIGMENTATION ON DESICCATION AND ULTRAVIOLET RESISTANCE IN TWO SPECIES OF DROSOPHILA

Drosophila yakuba and D. santomea are sister species that differ in their levels of abdominal pigmentation; D. yakuba shows heavily pigmented posterior abdominal segments in both sexes, whereas D. santomea lacks dark pigment anywhere on its body. Using naturally collected lines, we demonstrate the existence of altitudinal variation in abdominal pigmentation in D. yakuba but not in D. santomea. We use the variation in pigmentation within D. yakuba and two body‐color mutants in D. yakuba to elucidate selective advantage of differences in pigmentation. Our results indicate that although differences in abdominal pigmentation have no effect on desiccation resistance, lighter pigmentation confers ultraviolet radiation resistance in this pair of species.     

Regulation of 20-hydroxyecdysone on the larval pigmentation and the expression of melanin synthesis enzymes and yellow gene of the swallowtail butterfly, Papilio xuthus

The swallowtail butterfly, Papilio xuthus, changes its larval body pattern dramatically during the fourth ecdysis. Cuticular pigmentation occurs with precise timing just before ecdysis. We previously found that the cuticular pigmentation was regulated by three melanin synthesis genes, tyrosine hydroxylase (TH), dopa decarboxylase (DDC), and ebony. We discovered that yellow is strongly expressed in the presumptive black markings earlier than TH and DDC. Because the ecdysis is triggered by 20-hydroxyecdysone (20E), the effects of 20E on the pigmentation and expression of the melanin synthesis genes were examined. Here, we established a method for the topical application of 20E to molting specimens, so that 20E has only a partial effect, resulting in successful ecdysis. When we applied 20E during the mid-phase of the molting period, when the 20E titer is declining, cuticular pigmentation was completely inhibited. The cessation of hormonal treatments caused delayed pigmentation. yellow expression was promoted by a high titer of 20E, whereas the expression of TH, DDC, and ebony was suppressed, suggesting that a decline in the 20E concentration is necessary for the induction of the expression of the latter three genes. These results indicate that cuticular pigmentation is controlled by the exposure to 20E and its removal.     

Abdominal pigmentation variation in drosophila polymorpha: geographic variation in the trait, and underlying phylogeography

Drosophila polymorpha is a widespread species that exhibits abdominal pigmentation variation throughout its range. To gain insight into this variation we combined phenotypic and genotypic data to test a series of nested hypotheses. First, we tested the null hypothesis that geographic variation in pigmentation is due to neutral factors. We used nested clade analysis to examine the distribution of haplotypes from a nuclear and a mitochrondrial locus. Restricted gene flow via isolation by distance, the primary inference of this phylogeographic analysis, was then used to generate and test the hypothesis of increasing average abdominal pigmentation difference with increasing geographic distance. We found no correlation between geographic distance and phenotypic distance. We then tested the hypothesis that pigmentation is affected by environmental differences among localities. We found a significant effect of habitat type on the average abdominal pigmentation phenotype of different localities. Finally, we tested the hypothesis that pigmentation in D. polymorpha is associated with desiccation resistance. We found that dark individuals of both sexes survived significantly longer in a desiccating environment than light individuals. These patterns combined lead us to hypothesize that abdominal pigmentation variation in D. polymorpha is important in mediating the organism's interactions with local ecological factors.     

Natural variation of ebony gene controlling thoracic pigmentation in Drosophila melanogaster

We identified the causal genetic variation for the difference in the thoracic trident pigmentation intensity between two wild-derived strains of Drosophila melanogaster. It was found to be the difference in expression level of ebony, which codes for an enzyme in the melanin-synthesis pathway and has pleiotropic effects on vision and behavior.     

Genetic basis of natural variation in body pigmentation in Drosophila melanogaster

Body pigmentation in insects and other organisms is typically variable within and between species and is often associated with fitness. Regulatory variants with large effects at bab1, t and e affect variation in abdominal pigmentation in several populations of Drosophila melanogaster. Recently, we performed a genome wide association (GWA) analysis of variation in abdominal pigmentation using the inbred, sequenced lines of the Drosophila Genetic Reference Panel (DGRP). We confirmed the large effects of regulatory variants in bab1, t and e; identified 81 additional candidate genes; and validated 17 candidate genes (out of 28 tested) using RNAi knockdown of gene expression and mutant alleles. However, these analyses are imperfect proxies for the effects of segregating variants. Here, we describe the results of an extreme quantitative trait locus (xQTL) GWA analysis of female body pigmentation in an outbred population derived from light and dark DGRP lines. We replicated the effects on pigmentation of 28 genes implicated by the DGRP GWA study, including bab1, t and e and 7 genes previously validated by RNAi and/or mutant analyses. We also identified many additional loci. The genetic architecture of Drosophila pigmentation is complex, with a few major genes and many other loci with smaller effects.     

Genetic basis of natural variation in body pigmentation in Drosophila melanogaster

Body pigmentation in insects and other organisms is typically variable within and between species and is often associated with fitness. Regulatory variants with large effects at bab1, t and e affect variation in abdominal pigmentation in several populations of Drosophila melanogaster. Recently, we performed a genome wide association (GWA) analysis of variation in abdominal pigmentation using the inbred, sequenced lines of the Drosophila Genetic Reference Panel (DGRP). We confirmed the large effects of regulatory variants in bab1, t and e; identified 81 additional candidate genes; and validated 17 candidate genes (out of 28 tested) using RNAi knockdown of gene expression and mutant alleles. However, these analyses are imperfect proxies for the effects of segregating variants. Here, we describe the results of an extreme quantitative trait locus (xQTL) GWA analysis of female body pigmentation in an outbred population derived from light and dark DGRP lines. We replicated the effects on pigmentation of 28 genes implicated by the DGRP GWA study, including bab1, t and e and 7 genes previously validated by RNAi and/or mutant analyses. We also identified many additional loci. The genetic architecture of Drosophila pigmentation is complex, with a few major genes and many other loci with smaller effects.     

GROWTH TEMPERATURE AND ADULT PIGMENTATION IN TWO DROSOPHILA SIBLING SPECIES: AN ADAPTIVE CONVERGENCE OF REACTION NORMS IN SYMPATRIC POPULATIONS?

Phenotypic plasticity of abdomen pigmentation was investigated in populations of the sibling species Drosophila melanogaster and D. simulans, living in sympatry in two French localities. Ten isofemale lines of each population and species were grown at different constant temperatures spanning their complete thermal range from 12 to 31°C. Genetic variability between isofemale lines was not affected by growth temperature, but was consistently less in D. simulans. For all traits, the dark pigmentation of the abdominal segments decreased according to growth temperature, in agreement with the thermal budget adaptive hypothesis. The shapes of the response curves were different between the abdominal segments, but for a given segment, quite similar in the two species. On average D. simulans was lighter than D. melanogaster, but the difference was mainly expressed at higher temperatures. An interesting result was the difference observed between the two localities: flies from the colder locality (Villeurbanne) were found to be darker than flies from the warmer locality (Bordeaux). Interestingly, this difference was expressed only at low temperatures, 21°C and below, that is, at temperatures encountered in natural conditions. This suggests an adaptive response resulting in a change of the shape of reaction norm and involving genotype-environment interactions. When comparing the genetic structure of geographic populations for quantitative traits, several laboratory environments should be preferred to a single one.     

敲除ebony基因导致二化螟体表黑色素沉积

[目的]二化螟Chilo suppressalis是中国和其他亚洲地区为害水稻的重要害虫.由于缺乏遗传学操作工具,对二化螟的功能基因组学研究受到严重限制.本研究的目的是借助一个标记基因ebony,在二化螟体内建立CRISPR/Cas9介导的基因编辑体系.[方法]以家蚕Bombyx mori ebony蛋白作为参考序列,...     

黑腹果蝇黑条体突变体突变位点的鉴定

根据果蝇ebony基因的序列设计了一系列特异性引物,对1991年发现的一种黑腹果蝇的新的体色突变（Qian＆Zhang,1994）——黑条体突变型的ebony基因进行克隆和序列测定。与野生型W91910的ebony基因相比,黑条体突变型的ebony基因的编码区无明显的大片段变异,变异仅存在于数个氨基酸位点,且均不位于该基因产物的关键序列。在黑条体的ebony基因的5’端序列的克隆和序列测定中发现,与野生型w^91910及黑檀体e。的ebony基因相比,黑条体突变型的ebony基因有一个大片段的缺失,该缺失包括外显子1的206个碱基和内含子1的747个碱基,由此确定了黑条体突变体的突变类型和突变位点。     

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.     

A Genome-Wide,Fine-Scale Map of Natural Pigmentation Variation in Drosophila melanogaster

Various approaches can be applied to uncover the genetic basis of natural phenotypic variation, each with their specific strengths and limitations. Here, we use a replicated genome-wide association approach (Pool-GWAS) to fine-scale map genomic regions contributing to natural variation in female abdominal pigmentation in Drosophila melanogaster, a trait that is highly variable in natural populations and highly heritable in the laboratory. We examined abdominal pigmentation phenotypes in approximately 8000 female European D. melanogaster, isolating 1000 individuals with extreme phenotypes. We then used whole-genome Illumina sequencing to identify single nucleotide polymorphisms (SNPs) segregating in our sample, and tested these for associations with pigmentation by contrasting allele frequencies between replicate pools of light and dark individuals. We identify two small regions near the pigmentation genes tan and bric-à-brac 1, both corresponding to known cis-regulatory regions, which contain SNPs showing significant associations with pigmentation variation. While the Pool-GWAS approach suffers some limitations, its cost advantage facilitates replication and it can be applied to any non-model system with an available reference genome.     

An introgression approach to mapping differences in mating success and sperm competitive ability in Drosophila simulans and D. sechellia

The progeny of Drosophila females doubly-mated to males from the same and a closely related species are mostly sired by conspecific males. We examined the genetic basis for conspecific mating preference and sperm precedence by using 186 Drosophila lines in which random chromosomal fragments of D. sechellia were introgressed into D. simulans. Sperm competition was measured for each of these lines by crossing ebony D. simulans female with ebony D. simulans males followed by wild-type males from the introgressed lines. Variation in sperm competition (proportion of progeny sired by the second male), mating discrimination (proportion of introgressed males that failed to remate), and male fecundity (proportion of progeny sired by introgressed males) were scored. The introgressed lines exhibited highly significant heterogeneity in the three phenotypes scored, motivating an analysis to locate quantitative trait loci (QTLs) responsible for the differences. Applying composite interval mapping, we found eight QTLs that explain a significant level of variation among introgressed lines in the phenotypes scored. Cytological position overlapped among some QTLs suggesting possible pleiotropic effects. Analysis of the joint effects of simulans/sechellia genetic composition at different QTLs and markers suggests that complex interactions among alleles are partially responsible for interspecific differences in sexual traits.     

Local adaptation for body color in Drosophila americana

Pigmentation is one of the most variable traits within and between Drosophila species. Much of this diversity appears to be adaptive, with environmental factors often invoked as selective forces. Here, we describe the geographic structure of pigmentation in Drosophila americana and evaluate the hypothesis that it is a locally adapted trait. Body pigmentation was quantified using digital images and spectrometry in up to 10 flies from each of 93 isofemale lines collected from 17 locations across the United States and found to correlate most strongly with longitude. Sequence variation at putatively neutral loci showed no evidence of population structure and was inconsistent with an isolation-by-distance model, suggesting that the pigmentation cline exists despite extensive gene flow throughout the species range, and is most likely the product of natural selection. In all other Drosophila species examined to date, dark pigmentation is associated with arid habitats; however, in D. americana, the darkest flies were collected from the most humid regions. To investigate this relationship further, we examined desiccation resistance attributable to an allele that darkens pigmentation in D. americana. We found no significant effect of pigmentation on desiccation resistance in this experiment, suggesting that pigmentation and desiccation resistance are not unequivocally linked in all Drosophila species.     

Genetic Architecture of Abdominal Pigmentation in Drosophila melanogaster

Pigmentation varies within and between species and is often adaptive. The amount of pigmentation on the abdomen of Drosophila melanogaster is a relatively simple morphological trait, which serves as a model for mapping the genetic basis of variation in complex phenotypes. Here, we assessed natural variation in female abdominal pigmentation in 175 sequenced inbred lines of the Drosophila melanogaster Genetic Reference Panel, derived from the Raleigh, NC population. We quantified the proportion of melanization on the two most posterior abdominal segments, tergites 5 and 6 (T5, T6). We found significant genetic variation in the proportion of melanization and high broad-sense heritabilities for each tergite. Genome-wide association studies identified over 150 DNA variants associated with the proportion of melanization on T5 (84), T6 (34), and the difference between T5 and T6 (35). Several of the top variants associated with variation in pigmentation are in tan, ebony, and bric-a-brac1, genes known to affect D. melanogaster abdominal pigmentation. Mutational analyses and targeted RNAi-knockdown showed that 17 out of 28 (61%) novel candidate genes implicated by the genome-wide association study affected abdominal pigmentation. Several of these genes are involved in developmental and regulatory pathways, chitin production, cuticle structure, and vesicle formation and transport. These findings show that genetic variation may affect multiple steps in pathways involved in tergite development and melanization. Variation in these novel candidates may serve as targets for adaptive evolution and sexual selection in D. melanogaster.     

Five colour morphs and three new species of Gyptis (Hesionidae, Annelida) under a jetty in Edithburgh, South Australia

We report five different colour morphs of the hesionid polychaete genus Gyptis co-occurring in a small area in shallow water under Edithburgh jetty, South Australia. The five morphs cannot be separated using standard morphological features, but phylogenetic analyses of sequence data (COI and ITS1) unequivocally show that three species are present, introduced as Gyptis simpsonorum , new species, G. paucilineata , new species and G. polymorpha , new species. Gyptis simpsonorum has a speckled pigmentation pattern and G. paucilineata a few transverse lines on specific segments. Gyptis polymorpha is polymorphic with three different, distinct pigmentation patterns, either as dense transverse lines, as a thin longitudinal, mid-dorsal line, or as an uniformly dark dorsum. The speckled pigmentation likely represents the plesiomorphic condition, and G. propinqua is the closest known relative to these new species.     

Phenotypic plasticity of body pigmentation in Drosophila: influence of a developmental thermoperiodic regime in two sibling species

Abstract Variation of body pigmentation according to growth temperature was analysed in two European populations (Bordeaux and Cordoba) of the two sibling species Drosophila melanogaster (Meigen) and D. simulans (Sturtevant). Results obtained at constant temperatures were compared with those observed under alternating thermal regimes of various amplitudes and with a daily periodicity of 12–12 h. Abdominal pigmentation in segments 5, 6 and 7, and of their sum, was investigated in both species in females only. Thoracic trident pigmentation was studied in males and females but in Drosophila melanogaster only.
Thoracic pigmentation exhibited curvilinear convex reaction norms under constant conditions, with a darker pigmentation in the population (Bordeaux) living in a colder climate. No specific effects of alternating regimes were seen. For abdomen pigmentation, slightly different reaction norms were observed under constant conditions between Bordeaux and Cordoba strains. There were also slight differences between the two species. Alternating temperatures produced a significant decrease in average pigmentation, roughly proportional to the thermal amplitude.
Alternating stressing conditions induced also an increase in the phenotypic variability of abdominal pigmentation, especially in segments 6 and 7. Finally, because an increase of growth temperature results in a monotonically decreasing reaction norm of both abdomen pigmentation in females and a size-related trait, the wing/thorax ratio, we found a linear relationship between the two variables, with a very high overall correlation at constant temperatures ( r  = 0.99). The correlation was slightly less under alternating conditions, due to a broader dispersal of the experimental values around the regression lines.     

Fonction chromatrope de la pars intercerebralis chez l'acridien Oedipoda coerulescens

The median neurosecretory cells of the protocerebrum of fifth instar larvae and 1- or 2-day-old adults of Oedipoda coerulescens produced more fuchsinophilic material when reared on dark backgrounds than on pale ones. In the latter case, the neurosecretory cell bodies were almost completely devoid of neurosecretion. The same differences were found in the anterior lobes of the corpora cardiaca. Individuals implanted with pars intercerebralis from donors reared on a dark background showed dark pigmentation no matter what their own background colour (and conversely). Black or red pigmentation appeared on absorbent grounds whereas a pale one was produced on reflecting grounds with high levels of incident light predominantly between 576 and 588 mμ. The significance of these results is discussed. A possible scheme for the interaction of environmental factors and pigmentation is suggested.