Wolbachia Age-Sex-Specific Density in Aedes albopictus: A Host Evolutionary Response to Cytoplasmic Incompatibility?

Wolbachia bacteria have invaded many arthropod species by inducing Cytoplasmic Incompatibility (CI). These symbionts represent fascinating objects of study for evolutionary biologists, but also powerful potential biocontrol agents. Here, we assess the density dynamics of Wolbachia infections in males and females of the mosquito Aedes albopitcus, an important vector of human pathogens, and interpret the results within an evolutionary framework.Wolbachia densities were measured in natural populations and in age controlled mosquitoes using quantitative PCR. We show that the density dynamics of the wAlbA Wolbachia strain infecting Aedes albopictus drastically differ between males and females, with a very rapid decay of infection in males only.Theory predicts that Wolbachia and its hosts should cooperate to improve the transmission of infection to offspring, because only infected eggs are protected from the effects of CI. However, incompatible matings effectively lower the fertility of infected males, so that selection acting on the host genome should tend to reduce the expression of CI in males, for example, by reducing infection density in males before sexual maturation. The rapid decay of one Wolbachia infection in Aedes albopictus males, but not in females, is consistent with this prediction. We suggest that the commonly observed reduction in CI intensity with male age reflects a similar evolutionary process. Our results also highlight the importance of monitoring infection density dynamics in both males and females to assess the efficiency of Wolbachia-based control strategies

The evolution of sex ratio distorter suppression affects a 25 cM genomic region in the butterfly Hypolimnas bolina

Open Access ArticleSymbionts that distort their host's sex ratio by favouring the production and survival of females are common in arthropods. Their presence produces intense Fisherian selection to return the sex ratio to parity, typified by the rapid spread of host 'suppressor' loci that restore male survival/development. In this study, we investigated the genomic impact of a selective event of this kind in the butterfly Hypolimnas bolina. Through linkage mapping, we first identified a genomic region that was necessary for males to survive Wolbachia-induced male-killing. We then investigated the genomic impact of the rapid spread of suppression, which converted the Samoan population of this butterfly from a 100:1 female-biased sex ratio in 2001 to a 1:1 sex ratio by 2006. Models of this process revealed the potential for a chromosome-wide effect. To measure the impact of this episode of selection directly, the pattern of genetic variation before and after the spread of suppression was compared. Changes in allele frequencies were observed over a 25 cM region surrounding the suppressor locus, with a reduction in overall diversity observed at loci that co-segregate with the suppressor. These changes exceeded those expected from drift and occurred alongside the generation of linkage disequilibrium. The presence of novel allelic variants in 2006 suggests that the suppressor was likely to have been introduced via immigration rather than through de novo mutation. In addition, further sampling in 2010 indicated that many of the introduced variants were lost or had declined in frequency since 2006. We hypothesize that this loss may have resulted from a period of purifying selection, removing deleterious material that introgressed during the initial sweep. Our observations of the impact of suppression of sex ratio distorting activity reveal a very wide genomic imprint, reflecting its status as one of the strongest selective forces in nature.Natural Environment Research Council (NERC

Categorization of species as native or nonnative using DNA sequence signatures without a complete reference library.

New genetic diagnostic approaches have greatly aided efforts to document global biodiversity and improve biosecurity. This is especially true for organismal groups in which species diversity has been underestimated historically due to difficulties associated with sampling, the lack of clear morphological characteristics, and/or limited availability of taxonomic expertise. Among these methods, DNA sequence barcoding (also known as "DNA barcoding") and by extension, meta-barcoding for biological communities, has emerged as one of the most frequently utilized methods for DNA-based species identifications. Unfortunately, the use of DNA barcoding is limited by the availability of complete reference libraries (i.e., a collection of DNA sequences from morphologically identified species), and by the fact that the vast majority of species do not have sequences present in reference databases. Such conditions are critical especially in tropical locations that are simultaneously biodiversity rich and suffer from a lack of exploration and DNA characterization by trained taxonomic specialists. To facilitate efforts to document biodiversity in regions lacking complete reference libraries, we developed a novel statistical approach that categorizes unidentified species as being either likely native or likely nonnative based solely on measures of nucleotide diversity. We demonstrate the utility of this approach by categorizing a large sample of specimens of terrestrial insects and spiders (collected as part of the Moorea BioCode project) using a generalized linear mixed model (GLMM). Using a training data set of known endemic (n = 45) and known introduced species (n = 102), we then estimated the likely native/nonnative status for 4,663 specimens representing an estimated 1,288 species (412 identified species), including both those specimens that were either unidentified or whose endemic/introduced status was uncertain. Using this approach, we were able to increase the number of categorized specimens by a factor of 4.4 (from 794 to 3,497), and the number of categorized species by a factor of 4.8 from (147 to 707) at a rate much greater than chance (77.6% accuracy). The study identifies phylogenetic signatures of both native and nonnative species and suggests several practical applications for this approach including monitoring biodiversity and facilitating biosecurity

A DNA barcode-based survey of terrestrial arthropods in the Society Islands of French Polynesia : host diversity within the SymbioCode Project

We report here on the taxonomic and molecular diversity of 10 929 terrestrial arthropod specimens, collected on four islands of the Society Archipelago, French Polynesia. The survey was part of the 'SymbioCode Project' that aims to establish the Society Islands as a natural laboratory in which to investigate the flux of bacterial symbionts (e.g., Wolbachia) and other genetic material among branches of the arthropod tree. The sample includes an estimated 1127 species, of which 1098 included at least one DNA-barcoded specimen and 29 were identified to species level using morphological traits only. Species counts based on molecular data emphasize that some groups have been understudied in this region and deserve more focused taxonomic effort, notably Diptera, Lepidoptera and Hymenoptera. Some taxa that were also subjected to morphological scrutiny reveal a consistent match between DNA and morphology-based species boundaries in 90% of the cases, with a larger than expected genetic diversity in the remaining 10%. Many species from this sample are new to this region or are undescribed. Some are under description, but many await inspection by motivated experts, who can use the online images or request access to ethanol-stored specimens.Peer reviewe

The joint evolutionary histories of Wolbachia and mitochondria in Hypolimnas bolina

Background. The interaction between the Blue Moon butterfly, Hypolimnas bolina, and Wolbachia has attracted interest because of the high prevalence of male-killing achieved within the species, the ecological consequences of this high prevalence, the intensity of selection on the host to suppress the infection, and the presence of multiple Wolbachia infections inducing different phenotypes. We examined diversity in the co-inherited marker, mtDNA, and the partitioning of this between individuals of different infection status, as a means to investigate the population biology and evolutionary history of the Wolbachia infections. Results. Part of the mitochondrial COI gene was sequenced from 298 individuals of known infection status revealing ten different haplotypes. Despite very strong biological evidence that the sample represents a single species, the ten haplotypes did not fall within a monophyletic clade within the Hypolimnas genus, with one haplotype differing by 5% from the other nine. There were strong associations between infection status and mtDNA haplotype. The presence of wBol1 infection in association with strongly divergent haplotypes prompted closer examination of wBol1 genetic variation. This revealed the existence of two cryptic subtypes, wBol1a and wBol1b. The wBol1a infection, by far the most common, was in strict association with the single divergent mtDNA haplotype. The wBol1b infection was found with two haplotypes that were also observed in uninfected specimens. Finally, the wBol2 infection was associated with a large diversity of mtDNA haplotypes, most often shared with uninfected sympatric butterflies. Conclusion. This data overall supports the hypothesis that high prevalence of male-killing Wolbachia (wBol1) in H. bolina is associated with very high transmission efficiency rather than regular horizontal transmission. It also suggests this infection has undergone a recent selective sweep and was introduced in this species through introgression. In contrast, the sharing of haplotypes between wBol2-infected and uninfected individuals indicates that this strain is not perfectly transmitted and/or shows a significant level of horizontal transmission

Evolution of Male-Killer Suppression in a Natural Population

Male-killing bacteria are widespread in arthropods, and can profoundly alter the reproductive biology of their host species. Here we detail the first case of complete suppression of a male killer. The nymphalid butterfly Hypolimnas bolina is infected with a strain of the bacterium Wolbachia, wBol1, which kills male host embryos in Polynesian populations, but does not do so in many areas of Southeast Asia, where both males and female adults are naturally infected, and wBol1-infected females produce a 1:1 sex ratio. We demonstrate that absence of male killing by wBol1 is associated with dominant zygotic suppression of the action of the male killer. Simulations demonstrate host suppressors of male-killer action can spread very rapidly, and historical data indicating the presence of male killing in Southeast Asia in the very recent past suggests suppressor spread has been a very recent occurrence. Thus, male killer/host interactions are much more dynamic than previously recognised, with rapid and dramatic loss of the phenotype. Our results also indicate that suppression can render male killers completely quiescent, leading to the conclusion that some species that do not currently express a male killer may have done so in the past, and thus that more species have had their biology affected by these parasites than previously believed

The evolution of sex ratio distorter suppression affects a 25 cM genomic region in the butterfly Hypolimnas bolina.

Symbionts that distort their host's sex ratio by favouring the production and survival of females are common in arthropods. Their presence produces intense Fisherian selection to return the sex ratio to parity, typified by the rapid spread of host 'suppressor' loci that restore male survival/development. In this study, we investigated the genomic impact of a selective event of this kind in the butterfly Hypolimnas bolina. Through linkage mapping, we first identified a genomic region that was necessary for males to survive Wolbachia-induced male-killing. We then investigated the genomic impact of the rapid spread of suppression, which converted the Samoan population of this butterfly from a 100:1 female-biased sex ratio in 2001 to a 1:1 sex ratio by 2006. Models of this process revealed the potential for a chromosome-wide effect. To measure the impact of this episode of selection directly, the pattern of genetic variation before and after the spread of suppression was compared. Changes in allele frequencies were observed over a 25 cM region surrounding the suppressor locus, with a reduction in overall diversity observed at loci that co-segregate with the suppressor. These changes exceeded those expected from drift and occurred alongside the generation of linkage disequilibrium. The presence of novel allelic variants in 2006 suggests that the suppressor was likely to have been introduced via immigration rather than through de novo mutation. In addition, further sampling in 2010 indicated that many of the introduced variants were lost or had declined in frequency since 2006. We hypothesize that this loss may have resulted from a period of purifying selection, removing deleterious material that introgressed during the initial sweep. Our observations of the impact of suppression of sex ratio distorting activity reveal a very wide genomic imprint, reflecting its status as one of the strongest selective forces in nature.This is the final version of the article. It first appeared from PLOS via http://dx.doi.org/10.1371/journal.pgen.100482

Draft genome sequence of the male-killing Wolbachia strain wBol1 reveals recent horizontal gene transfers from diverse sources.

Background The endosymbiont Wolbachia pipientis causes diverse and sometimes dramatic phenotypes in its invertebrate hosts. Four Wolbachia strains sequenced to date indicate that the constitution of the genome is dynamic, but these strains are quite divergent and do not allow resolution of genome diversification over shorter time periods. We have sequenced the genome of the strain wBol1-b, found in the butterfly Hypolimnas bolina, which kills the male offspring of infected hosts during embyronic development and is closely related to the non-male-killing strain wPip from Culex pipiens. Results The genomes of wBol1-b and wPip are similar in genomic organisation, sequence and gene content, but show substantial differences at some rapidly evolving regions of the genome, primarily associated with prophage and repetitive elements. We identified 44 genes in wBol1-b that do not have homologs in any previously sequenced strains, indicating that Wolbachia's non-core genome diversifies rapidly. These wBol1-b specific genes include a number that have been recently horizontally transferred from phylogenetically distant bacterial taxa. We further report a second possible case of horizontal gene transfer from a eukaryote into Wolbachia. Conclusions Our analyses support the developing view that many endosymbiotic genomes are highly dynamic, and are exposed and receptive to exogenous genetic material from a wide range of sources. These data also suggest either that this bacterial species is particularly permissive for eukaryote-to-prokaryote gene transfers, or that these transfers may be more common than previously believed. The wBol1-b-specific genes we have identified provide candidates for further investigations of the genomic bases of phenotypic differences between closely-related Wolbachia strains.Peer reviewe

Les contours flous de l’individu

L’individu, souvent, nous saute aux yeux : vous et moi, mon chat, cet oiseau qui passe... Mais l’évidence, parfois, se dérobe. Ce champignon, ou son corps sous-terrain ? Ce peuplier, ou tous ses clones, cette forêt tout entière ? Cette bactérie, ou ce biofilm ? Cette fourmi stérile, ce petit organe à six pattes, ou sa fourmilière ? Ce puceron, et ce symbiote intracellulaire, ou la nouvelle unité qu’ils composent ? Vous et moi, ou ces éléments hétéroclites, et pour la plupart égoïstes, qui constituent nos génomes ? Ou nos innombrables cellules, et la ménagerie de notre microbiote ? À la lumière de la symbiose, du conflit et de la coopération, l’histoire du vivant devient une histoire d’associations et de dissociations, une histoire de changements d’échelles. Les cellules s’assemblent pour former des organismes multicellulaires, où l’hérédité est réglée à la baguette par les strictes lois de Mendel, mais les occasions de s’y dérober ne manquent pas. De nouvelles échelles émergent, par association, sans que les échelles antérieures ne disparaissent tout à fait. De nouveaux « niveaux de sélection » prennent le contrôle, de nouveaux « individus » émergent, mais la reproduction sexuée ou le transfert horizontal sont autant d’occasions pour les éléments qui les composent d’exister encore « pour eux-mêmes », de constituer eux aussi des niveaux de sélection efficaces, et à ce titre, des échelles de description pertinentes pour « comprendre » le vivant, pour révéler, derrière les structures, des fonctions. Mes travaux de recherche s’inscrivent dans cette perspective, dans ces questionnements. Ils apportent parfois des bribes de réponses mais consistent essentiellement (je le constate à travers ce manuscrit) à questionner les questions elles-mêmes, à les inspecter sous tous les angles. Ils portent sur la symbiose, sur Wolbachia, dont les stratégies d’invasion égoïstes défient l’imagination, même si ces bactéries intracellulaires sont parfois essentielles, si elles peuvent devenir de leur hôte le « meilleur ennemi ». Ils portent sur le transfert horizontal, cette transmission de matériel génétique entre différentes lignées évolutives, notamment entre différentes espèces d’insectes, des papillons et leur parasitoïdes, ces prédateurs de l’intérieur, ces Aliens. Mes travaux portent aussi sur la nature de l’hérédité, sur les possibles limitations d’un modèle, celui de la génétique, dont la puissance explicative pourrait masquer d’autres formes d’hérédité, peut-être trop instables pour peser lourd dans l’évolution, mais peut-être pas. Enfin, mes travaux portent sur une hypothèse, celle d’une évolution biologique qui dépasserait ses frontières, d’une sélection naturelle qui pourrait être à l’œuvre en dehors du vivant que nous connaissons, dans d’autres systèmes physiques ou chimiques, à d’autres échelles temporelles ou spatiales, en d’autres lieux, ou peut-être ici même... mais serions-nous seulement capables de les reconnaître ? Pour l’heure, cette exploration se heurte à un obstacle, un problème de poule et d’œuf où le concept d’individu se fait à nouveau central : sans individus, pas de variation héritable, pas de sélection naturelle, pas d’évolution ; mais sans évolution, pas d’adaptations, pas de « parties fonctionnelles d’un tout », pas d’unités de sélection, pas d’individus... Quelle forme pourrait prendre une (méta)théorie de l’évolution qui échapperait à ce paradoxe, une théorie évolutive « de l’évolution elle-même », qui traverserait la biologie et la physique, en expliquant, sans s’y référer, l’émergence de l’individu