Introgression in hybrid ants is favored in females but selected against in males.

Hybridization is not a mere reproductive dead end but has been suggested to play a central role in speciation, for example, by introducing adaptive genetic variation. Our previous study uncovered a unique consequence of hybridization in Formica ants. In a population including two isolated but partially introgressed genetic groups, the females have an apparent hybrid background, whereas the males do not. This situation results in large-scale differences between male and female genomes that are stable throughout generations. Here, we compare genotypes from different developmental stages to investigate how sex-specific introgression and genetic differences between sexes are maintained. We show that strong selection rather than sex-dependent transmission maintains the genetic differences between sexes. All genotype combinations are produced and observed in the eggs of both sexes, but the alleles acquired through hybridization disappear from the haploid males during development from egg to adult as their frequencies drop toward zero. However, the same introgressed alleles increase in frequency and are favored when heterozygous in the females. Genotypes eliminated from males most likely represent incompatibilities arising from hybridization. Our results show an unusual situation of opposite selection, where introgression is favored in diploid females but selected against in haploid males. This finding suggests that introgressed genomic regions harbor both fitness-enhancing and -reducing elements. Our work highlights the complex consequences of hybridization and provides a rare opportunity to observe natural selection in real time in nature.

The genome of the fire ant Solenopsis invicta.

Ants have evolved very complex societies and are key ecosystem members. Some ants, such as the fire ant Solenopsis invicta, are also major pests. Here, we present a draft genome of S. invicta, assembled from Roche 454 and Illumina sequencing reads obtained from a focal haploid male and his brothers. We used comparative genomic methods to obtain insight into the unique features of the S. invicta genome. For example, we found that this genome harbors four adjacent copies of vitellogenin. A phylogenetic analysis revealed that an ancestral vitellogenin gene first underwent a duplication that was followed by possibly independent duplications of each of the daughter vitellogenins. The vitellogenin genes have undergone subfunctionalization with queen- and worker-specific expression, possibly reflecting differential selection acting on the queen and worker castes. Additionally, we identified more than 400 putative olfactory receptors of which at least 297 are intact. This represents the largest repertoire reported so far in insects. S. invicta also harbors an expansion of a specific family of lipid-processing genes, two putative orthologs to the transformer/feminizer sex differentiation gene, a functional DNA methylation system, and a single putative telomerase ortholog. EST data indicate that this S. invicta telomerase ortholog has at least four spliceforms that differ in their use of two sets of mutually exclusive exons. Some of these and other unique aspects of the fire ant genome are likely linked to the complex social behavior of this species.

Towards greater realism in inclusive fitness models: the case of worker reproduction in insect societies.

The conflicts over sex allocation and male production in insect societies have long served as an important test bed for Hamilton's theory of inclusive fitness, but have for the most part been considered separately. Here, we develop new coevolutionary models to examine the interaction between these two conflicts and demonstrate that sex ratio and colony productivity costs of worker reproduction can lead to vastly different outcomes even in species that show no variation in their relatedness structure. Empirical data on worker-produced males in eight species of Melipona bees support the predictions from a model that takes into account the demographic details of colony growth and reproduction. Overall, these models contribute significantly to explaining behavioural variation that previous theories could not account for.

Segregation distortion causes large-scale differences between male and female genomes in hybrid ants.

Hybridization in isolated populations can lead either to hybrid breakdown and extinction or in some cases to speciation. The basis of hybrid breakdown lies in genetic incompatibilities between diverged genomes. In social Hymenoptera, the consequences of hybridization can differ from those in other animals because of haplodiploidy and sociality. Selection pressures differ between sexes because males are haploid and females are diploid. Furthermore, sociality and group living may allow survival of hybrid genotypes. We show that hybridization in Formica ants has resulted in a stable situation in which the males form two highly divergent gene pools whereas all the females are hybrids. This causes an exceptional situation with large-scale differences between male and female genomes. The genotype differences indicate strong transmission ratio distortion depending on offspring sex, whereby the mother transmits some alleles exclusively to her daughters and other alleles exclusively to her sons. The genetic differences between the sexes and the apparent lack of multilocus hybrid genotypes in males can be explained by recessive incompatibilities which cause the elimination of hybrid males because of their haploid genome. Alternatively, differentiation between sexes could be created by prezygotic segregation into male-forming and female-forming gametes in diploid females. Differentiation between sexes is stable and maintained throughout generations. The present study shows a unique outcome of hybridization and demonstrates that hybridization has the potential of generating evolutionary novelties in animals.

A high recombination rate in eusocial Hymenoptera: evidence from the common wasp Vespula vulgaris.

BACKGROUND: High recombination rates have previously been detected in two groups of eusocial insects; honeybees and ants. In this study we estimate recombination rate in a eusocial wasp Vespula vulgaris that represents a third phylogenetic lineage within eusocial hymenopterans. RESULTS: A genetic linkage map of V. vulgaris based on 210 markers shows that the total map length is 2129 cM and the recombination rate is 9.7 cM/Mb (or 103 kb/cM). The present estimate in V. vulgaris is somewhat smaller than in the honeybee Apis mellifera and intermediate between the estimates from two ant species (Acromyrmex echinatior, Pogonomyrmex rugosus). Altogether, the estimates from these eusocial species are higher than in any other insect reported so far. CONCLUSIONS: [corrected] The four species (V. vulgaris, A. mellifera, A. echinatior, P. rugosus) are characterized by advanced eusociality with large colonies, clear queen-worker dimorphism and well developed task specialization. They also have colonies with a single, normally multiply inseminated (polyandrous) queen. Benefits of genotypic diversity within colonies (e.g. through improved task specialization or pathogen and parasite resistance) may have selected for both polyandry and high recombination rate in such advanced eusocial insects.

Multiple endosymbionts in populations of the ant Formica cinerea.

BACKGROUND: Many insects, including ants, are infected by maternally inherited Wolbachia endosymbiotic bacteria though other secondary endosymbionts have not been reported in ants. It has been suggested that the ability of Wolbachia to invade and remain in an ant population depends on the number of coexisting queens in a colony. We study the genetic and social structure of populations in the ant Formica cinerea which is known to have populations with either monogynous or polygynous colonies. We screen populations for several endosymbiotic bacteria to evaluate the presence of different endosymbionts, possible association between their prevalence and the social structure, and the association between endosymbiont prevalence and genetic differentiation of ant populations. RESULTS: We found three endosymbiotic bacteria; 19% of the nests were infected by Wolbachia, 3.8% by Cardinium and 33% by Serratia. There was significant variation among the populations regarding the proportion of nests infected by Serratia, Wolbachia and the pooled set of all the endosymbionts. Some individuals and colonies carried two of the bacteria, the frequency of double infections agreeing with the random expectation. The proportion of infected ants (individuals or colonies) did not correlate significantly with the population level relatedness values. The difference in the prevalence of Wolbachia between population pairs correlated significantly with the genetic distance (microsatellites) of the populations. CONCLUSIONS: The discovery of several endosymbionts and co-infections by Wolbachia and Cardinium demonstrate the importance of screening several endosymbionts when evaluating their possible effects on social life and queen-worker conflicts over sex allocation. The low prevalence of Wolbachia in F. cinerea departs from the pattern observed in many other Formica ants in which all workers have been infected. It is likely that the strain of Wolbachia in F. cinerea differs from those in other Formica species. The correlation between the difference in Wolbachia prevalence and the pair-wise genetic distance of populations suggests that spreading of the bacteria is restricted by the isolation of the host populations.

Rapid evolution of immune proteins in social insects.

The existence of behavioral traits connected to defense against pathogens manifests the importance of pathogens in the evolution of social insects. However, very little is known about how pathogen pressure has affected the molecular evolution of genes involved in their innate immune system. We have studied the sequence evolution of several immune genes in ants and honeybees. The results show high rates of evolution in both ants and honeybees as measured by the ratio of amino acid changes to silent nucleotide changes, the ratio being clearly higher than in Drosophila immune genes or in nonimmunity genes of bees. This conforms to our expectations based on high pathogen pressure in social insects. The codon-based likelihood method found clear evidence of positive selection only in one ant gene, even though positive selection has earlier been found in both ant and termite immune genes. There is now indication that selection on the amino acid composition of the immune-related genes has been an important part in the fight against pathogens by social insects. However, we cannot distinguish in all the cases whether the high observed d(N)/d(S) ratio results from positive selection within a restricted part of the studied genes or from relaxation of purifying selection associated with effective measures of behaviorally based colony-level defenses.

Wolbachia transmission dynamics in Formica wood ants.

BACKGROUND: The role of Wolbachia endosymbionts in shaping the mitochondrial diversity of their arthropod host depends on the effects they have on host reproduction and on the mode of transmission of the bacteria. We have compared the sequence diversity of wsp (Wolbachia surface protein gene) and the host mtDNA in a group of Formica ant species that have diverged approximately 0.5 million years ago (MYA). The aim was to study the relationship of Wolbachia and its ant hosts in terms of vertical and horizontal transmission of the bacteria. RESULTS: All studied ant species were doubly infected with two Wolbachia strains (wFex1 and wFex4) all over their geographical distribution area in Eurasia. The most common haplotypes of these strains were identical with strains previously described from a more distantly related Formica ant, with an estimated divergence time of 3.5 - 4 MYA. Some strain haplotypes were associated to the same or closely related mtDNA haplotypes as expected under vertical transmission. However, in several cases the wsp haplotypes coexisted with distant mtDNA haplotypes, a pattern which is more compatible with horizontal transmission of the bacteria. CONCLUSION: Two lines of evidence suggest that the sharing of Wolbachia strains by all F. rufa species is rather due to horizontal than vertical transmission. First, the fact that endosymbiont strains identical to those of F. rufa ants have been found in another species that diverged 3.5-4 MYA strongly suggests that horizontal transfer can and does occur between Formica ants. Second, the frequent sharing of identical Wolbachia strains by distant mitochondrial lineages within the F. rufa group further shows that horizontal transmission has occurred repeatedly. Nevertheless, our dataset also provides some evidence for longer-term persistence of infection, indicating that Wolbachia infection within this host clade has been shaped by both horizontal and vertical transmission of symbionts. The fact that all the ants were infected irrespective of the family structure of their societies gives no support to the proposed hypotheses that the spreading of Wolbachia in ants might be associated to the types of their societies.

Sex allocation conflict between queens and workers in Formica pratensis wood ants predicts seasonal sex ratio variation.

Sex allocation theory predicts parents should adjust their investment in male and female offspring in a way that increases parental fitness. This has been shown in several species and selective contexts. Yet, seasonal sex ratio variation within species and its underlying causes are poorly understood. Here, we study sex allocation variation in the wood ant Formica pratensis. This species displays conflict over colony sex ratio as workers and queens prefer different investment in male and female offspring, owing to haplodiploidy and relatedness asymmetries. It is unique among Formica ants because it produces two separate sexual offspring cohorts per season. We predict sex ratios to be closer to queen optimum in the early cohort but more female-biased and closer to worker optimum in the later one. This is because the power of workers to manipulate colony sex ratio varies seasonally with the availability of diploid eggs. Consistently, more female-biased sex ratios in the later offspring cohort over a three-year sampling period from 93 colonies clearly support our prediction. The resulting seasonal alternation of sex ratios between queen and worker optima is a novel demonstration how understanding constraints of sex ratio adjustment increases our ability to predict sex ratio variation.

Larger genetic differences within africans than between Africans and Eurasians.

The worldwide pattern of single nucleotide polymorphism (SNP) variation is of great interest to human geneticists, population geneticists, and evolutionists, but remains incompletely understood. We studied the pattern in noncoding regions, because they are less affected by natural selection than are coding regions. Thus, it can reflect better the history of human evolution and can serve as a baseline for understanding the maintenance of SNPs in human populations. We sequenced 50 noncoding DNA segments each approximately 500 bp long in 10 Africans, 10 Europeans, and 10 Asians. An analysis of the data suggests that the sampling scheme is adequate for our purpose. The average nucleotide diversity (pi) for the 50 segments is only 0.061% +/- 0.010% among Asians and 0.064% +/- 0.011% among Europeans but almost twice as high (0.115% +/- 0.016%) among Africans. The African diversity estimate is even higher than that between Africans and Eurasians (0.096% +/- 0.012%). From available data for noncoding autosomal regions (total length = 47,038 bp) and X-linked regions (47,421 bp), we estimated the pi-values for autosomal regions to be 0.105, 0.070, 0.069, and 0.097% for Africans, Asians, Europeans, and between Africans and Eurasians, and the corresponding values for X-linked regions to be 0.088, 0.042, 0.053, and 0.082%. Thus, Africans differ from one another slightly more than from Eurasians, and the genetic diversity in Eurasians is largely a subset of that in Africans, supporting the out of Africa model of human evolution. Clearly, one must specify the geographic origins of the individuals sampled when studying pi or SNP density.

Coexistence of the social types: genetic population structure in the ant Formica exsecta.

The ant Formica exsecta has two types of colonies that exist in sympatry but usually as separate subpopulations: colonies with simple social organization and single queens (M type) or colonial networks with multiple queens (P type). We used both nuclear (DNA microsatellites) and mitochondrial markers to study the transition between the social types, and the contribution of males and females in gene flow within and between the types. Our results showed that the social types had different spatial genetic structures. The M subpopulations formed a fairly uniform population, whereas the P subpopulations were, on average, more differentiated from each other than from the nearby M subpopulations and could have been locally established from the M-type colonies, followed by philopatric behavior and restricted emigration of females. Thus, the relationship between the two social types resembles that of source (M type) and sink (P type) populations. The comparison of mitochondrial (phiST) and nuclear (FST) differentiation indicates that the dispersal rate of males is four to five times larger than that of females both among the P-type subpopulations and between the social types. Our results suggest that evolution toward complex social organization can have an important effect on genetic population structure through changes in dispersal behavior associated with different sociogenetic organizations.

Phylogenetic relationships of Palaearctic Formica species (Hymenoptera, Formicidae) based on mitochondrial cytochrome B sequences.

Ants of genus Formica demonstrate variation in social organization and represent model species for ecological, behavioral, evolutionary studies and testing theoretical implications of the kin selection theory. Subgeneric division of the Formica ants based on morphology has been questioned and remained unclear after an allozyme study on genetic differentiation between 13 species representing all subgenera was conducted. In the present study, the phylogenetic relationships within the genus were examined using mitochondrial DNA sequences of the cytochrome b and a part of the NADH dehydrogenase subunit 6. All 23 Formica species sampled in the Palaearctic clustered according to the subgeneric affiliation except F. uralensis that formed a separate phylogenetic group. Unlike Coptoformica and Formica s. str., the subgenus Serviformica did not form a tight cluster but more likely consisted of a few small clades. The genetic distances between the subgenera were around 10%, implying approximate divergence time of 5 Myr if we used the conventional insect divergence rate of 2% per Myr. Within-subgenus divergence estimates were 6.69% in Serviformica, 3.61% in Coptoformica, 1.18% in Formica s. str., which supported our previous results on relatively rapid speciation in the latter subgenus. The phylogeny inferred from DNA sequences provides a necessary framework against which the evolution of social traits can be compared. We discuss implications of inferred phylogeny for the evolution of social traits.

Origin and evolution of the dependent lineages in the genetic caste determination system of Pogonomyrmex ants.

Hybridizing harvester ants of the Pogonomyrmex barbatus/rugosus complex have an exceptional genetic caste determination (GCD) mechanism. We combined computer simulations, population genomics, and linkage mapping using >1000 nuclear AFLP markers and a partial mtDNA sequence to explore the genetic architecture and origin of the dependent lineages. Our samples included two pairs of hybridizing lineages, and the mitochondrial and nuclear data showed contradicting affinities between them. Clustering of individual genotypes based on nuclear markers indicated some exceptions to the general GCD system, that is, interlineage hybrid genes as well as some pure-line workers. A genetic linkage map of P. rugosus showed one of the highest recombination rates ever measured in insects (14.0 cM/Mb), supporting the view that social insects are characterized by high recombination rates. The population data had 165 markers in which sibling pairs showed a significant genetic difference depending on the caste. The differences were scattered in the genome; 13 linkage groups had loci with F(ST)>0.9 between the hybridizing lineages J1 and J2.The mapping results and the population data indicate that the dependent lineages have been initially formed through hybridization at different points in time but the role of introgression has been insignificant in their later evolution.

Selection on an antimicrobial peptide defensin in ants.

Ants live in crowded nests with interacting individuals, which makes them particularly prone to infectious diseases. The question is, how do ants cope with the increased risk of pathogen transmission due to sociality? We have studied the molecular evolution of defensin, a gene encoding an antimicrobial protein, in ants. Defensin sequences from several ant species were analyzed with maximum likelihood models of codon substitution to infer selection. Positive selection was detected in the mature region of defensin, whereas the signal and pro regions seem to be evolving neutrally. We also found a significantly higher rate of nonsynonymous substitutions in some phylogenetic lineages, as well as dN/dS >1, suggesting varying selection pressures in different lineages. Earlier studies on the molecular evolution of insect antimicrobial peptide genes have focused on termites and dipteran species, and detected positive selection only in duplicated termicin genes in termites. These findings, together with our present results, provide an indication that the immune systems of social insects (ants and termites) and dipteran insects may have responded differently to the selection pressure caused by microbial pathogens.

Exceptionally high density of NUMTs in the honeybee genome.

The available genome sequences of 4 insects (the fruit fly, the African malaria mosquito, the flour beetle, and the honeybee) are used to compare the amount of mitochondrial DNA transferred to the nuclear genome (NUMTs). The data from the beetle and the bee show frequent transfer of NUMTs, whereas NUMTs in the 2 other insects are rare. The density of NUMTs in the honeybee (>1.0 bp transferred DNA per 1 kb of the nuclear sequence) is the highest in any animal studied, about ten times higher than in humans and comparable to the densities in plant genomes. The density of NUMTs in the beetle (0.056 bp/kb) is of the same order of magnitude as that in humans. The analysis of the honeybee genome indicates that NUMTs originate from all parts of the mitochondrial genome, that about two-thirds of the nuclear copies result from secondary transpositions within the nuclear genome, that the copies are significantly associated to "mariner" type transposons, and that the NUMTs consist mainly of short and fragmented copies.

STATISTICAL TESTS OF PHENOGRAMS BASED ON GENETIC DISTANCES.

Methods used for estimating the confidence levels of UPGMA-type phenograms based on molecular-genetic data are examined. The methods based on internodal variances or on bootstrapping over characters are compared by simulating a three-species phylogeny with a trifurcation. The bootstrap method seems slightly better in this comparison. Weighting of OTU's when constructing the phenogram is also examined. A method that weights each OTU according to the estimated independent evolutionary information, a modified WPGMA, appears slightly better than UPGMA in estimating the branching points and branch lengths. The methods are applied to the data on restriction sites in eight Hawaiian Drosophila species. The significance of the clusters among them differ from those reported earlier.

Highly variable social organisation of colonies in the ant Formica cinerea.

Social organisation of colonies was examined in the ant Formica cinerea by estimating the coefficient of genetic relatedness among worker nest mates. The estimates based on microsatellite genotypes at three loci ranged from values close to zero to 0.61 across the populations studied in Finland. These results showed that a fundamental feature of colonies, the number of reproductive queens, varied greatly among the populations. Colonies in some populations had a single queen, whereas the nests could have a high number number of queens in other populations. There was a weak but non-significant correlation between the genetic and metric distance of nests within two populations with intermediate level of relatedness. Differentiation among nearby populations (within the dispersal distance of individuals) in one locality indicated limited dispersal or founder effects. This could occur when females are philopatric and stay in the natal polygynous colony which expands by building a network of nest galleries within a single habitat patch.

Recent speciation in the Formica rufa group ants (Hymenoptera, Formicidae): inference from mitochondrial DNA phylogeny.

This study examines phylogenetic relationships among six species of the Formica rufa group ants (F. polyctena, F. rufa, F. lugubris, F. paralugubris, F. aquilonia, and F. pratensis). The phylogeny based on a 2051bp fragment of mtDNA including cyt b, tRNASer, and ND1 genes supports the division of the group into three major clusters: one with the species F. polyctena and F. rufa, one with F. aquilonia, F. lugubris, and F. paralugubris, and the third one with F. pratensis. The interspecific divergence estimates (mean 0.98 +/- 0.15% for the main phylogenetic groups) imply that radiation took place during the Pleistocene. Comparison of the divergence estimates among the F. rufa group species with divergence estimates among other closely related species of insects suggests that speciation in the group was relatively fast, and the mitochondrial lineages of F. polyctena and F. rufa have not fully separated. The haplotype tree shows also signs of transfer of mtDNA between species through hybridisation. The distribution of polygyny (multiple queens per nest) along the branches of the tree indicates that the social type characterised by highly polygynous societies and large colonial networks, has originated at least three times. The species F. aquilonia and F. paralugubris that build such large supercolonies, cluster tightly together with very little nucleotide variation, suggesting that this type of social organisation could be a factor promoting speciation in the ants.