Small-scale spatial genetic structure in an ant species with sex-biased dispersal

In a population of the monogynous, polyandrous ant Cataglyphis cursor , we analysed the spatial genetic structure of queens, colony fathers and workers at a microgeographical scale to infer the extent of sex-biased dispersal and to assess the impact of limited dispersal on the patterns of relatedness within the colony. To this end, four microsatellite markers were scored for the queen and an average of 26 workers from each of 35 mapped colonies. We used pair-wise kinship coefficients between all pairs of genotypes, including the reconstructed colony father genotypes (1) to test and quantify isolation by distance patterns within each sex or caste through the analysis of kinship–distance curves, and (2) to compute the average relatedness between categories of colony members. The kinship–distance curve was much steeper for colony queens than colony fathers, indicating male-biased dispersal. However, colony fathers also displayed a non-random spatial genetic structure, so that even males show some dispersal limitation at the scale of the population, which extends over less than 250 m. The degree of relatedness between the different sexes and castes of colonies was well predicted from the number of mates per queen and the inbreeding of queens, and the impact of limited dispersal was very weak at this scale of observation. We discuss the interest of kinship–distance curves to assess sex-biased dispersal on a local scale and we compare our results with large-scale analyses of genetic structure in Cataglyphis cursor and other monogynous ant species.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 93 , 465–473.     

Habitat age, breeding system and kinship in the ant Formica fusca

In polygyne ants (multiple queens per colony) factors that affect the distribution and survival of queens may play a key role in shaping the population-wide mating system and colony kin structure. The aim of this paper was to study the breeding system in two populations of different age in the facultatively polygyne ant Formica fusca. Both the observed numbers of queens, and the relatedness patterns among queens, workers and colony fathers were compared in two adjacent populations (ages 17 years and > 100 years) in Southern Finland. The results showed that both the mating system and colony kin structure differed between the study populations. In the old population the relatedness among workers, queens and colony fathers was high. The queens were also related to their mates, resulting in significant inbreeding in workers, but not in queens. Finally, the number of queens per colony fluctuated between years, suggesting queen turnover, and nest-mate queens shared their reproduction unequally (reproductive skew). In the younger population relatedness among queens and workers was lower than in the old population, and the colony fathers were unrelated. Furthermore, inbreeding was absent, and no conclusive evidence was found for reproductive skew among nest-mate queens. Finally, the number of queens per colony appeared more stable between years, although queen turnover occurred also in this population. The observed differences in dispersal and mating behaviour are discussed in the light of a potential connection between population age and habitat saturation.     

Male-biased dispersal promotes large scale gene flow in a subterranean army ant, Dorylus (Typhlopone) fulvus

Sex-biased dispersal is a widespread phenomenon in the animal kingdom, which strongly influences gene flow and population structure. Particularly army ants, important key-stone predators in tropical ecosystems, are prone to population fragmentation and isolation due to their extraordinary mating system: queens are permanently wingless, propagate via colony fission, and only the males disperse in mating flights. Here we report on sex-biased dispersal and the genetic population structure of an African subterranean army ant, Dorylus (Typhlopone) fulvus. Using maternally inherited mtDNA markers and bi-parentally inherited nuclear microsatellites we found strong geographical structuring of mtDNA haplotypes, whereas the nuclear genetic population structure was less pronounced. Strong mtDNA (Φ _ST = 0.85), but significantly lower nuclear (F _ST = 0.23) genetic differentiation translated into a more than an order of magnitude larger male migration rate compared to that of queens, reflecting the low motility of queens and strong, promiscuous dispersal by males. Thus, the well flying D. fulvus males appear to be the sex to promote large scale gene flow, and D. fulvus is indeed a species in which sex specific dispersal patterns and the mating system profoundly affect the population structure and phylogeography.     

Inbreeding and kinship in the ant Plagiolepis pygmaea

In ants the presence of multiple reproductive queens (polygyny) decreases the relatedness among workers and the brood they rear, and subsequently dilutes their inclusive fitness benefits from helping. However, adoption of colony daughters, low male dispersal in conjunction with intranidal (within nest) mating and colony reproduction by budding may preserve local genetic differences, and slow down the erosion of relatedness. Reduced dispersal and intranidal mating may, however, also lead to detrimental effects owing to competition and inbreeding. We studied mating and dispersal patterns, and colony kinship in three populations of the polygynous ant Plagiolepis pygmaea using microsatellite markers. We found that the populations were genetically differentiated, but also a considerable degree of genetic structuring within populations. The genetic viscosity within populations can be attributed to few genetically homogeneous colony networks, which presumably have arisen through colony reproduction by budding. Hence, selection may act at different levels, the individuals, the colonies and colony networks. All populations were also significantly inbred (F=0.265) suggesting high frequencies of intranidal mating and low male dispersal. Consequently the mean regression relatedness among workers was significantly higher (r = 0.529-0.546) than would be expected under the typically reported number (5-35) of queens in nests of the species. Furthermore, new queens were mainly recruited from their natal or a neighbouring related colony. Finally, the effective number of queens coincided with that found upon excavation, suggesting low reproductive skew.     

Population structure and mating biology of the polygynous ponerine ant Gnamptogenys striatula in Brazil

Gnamptogenys striatula is a polygynous ponerine ant, whose colonies contain either several differentiated queens or several gamergates. Population structure, queen mating frequency and deviation from random mating were investigated in a north-eastern Brazilian population. Eight workers from each of 33 queenright colonies and 17 queens and their progeny (20-40 offspring) were genotyped using eight variable microsatellite markers. Population differentiation tests indicated limited gene flow at the scale of several kilometres, and tests of isolation by distance revealed population viscosity at the scale of a few metres. This population structure, together with the frequent colony migrations and fissions observed in the field, suggest that new nests are founded by budding in G. striatula. Genetic data showed that 13 of our 17 queens were single-mated and four were double-mated. The estimation of the range of maximal frequency of double-mated queens in the population was 0.232-0.259, demonstrating that mating frequency is low in G. striatula. The low estimated mean relatedness between the 17 queens and their mates (-0. 04 +/- 0.49) indicated no evidence of inbreeding in G. striatula.     

Sib‐mating in the ant Plagiolepis pygmaea: adaptative inbreeding?

Multiple functional queens in a colony (polygyny) and multiple mating by queens (polyandry) in social insects challenge kin selection, because they dilute inclusive fitness benefits from helping. Colonies of the ant Plagiolepis pygmaea brash contain several hundreds of multiply mated queens. Yet, within‐colony relatedness remains unexpectedly high. This stems from low male dispersal, extensive mating among relatives and adoption of young queens in the natal colony. We investigated whether inbreeding results from workers expelling foreign males, and/or from preferential mating between related partners. Our data show that workers actively repel unrelated males entering their colony, and that queens preferentially mate with related males. These results are consistent with inclusive fitness being a driving force for inbreeding: by preventing outbreeding, workers reduce erosion of relatedness within colonies due to polygyny and polyandry. That virgin queens mate preferentially with related males could result from a long history of inbreeding, which is expected to reduce depression in species with regular sibmating.     

Polygyny and polyandry in small ant societies

Social insects, ants in particular, show considerable variation in queen number and mating frequency resulting in a wide range of social structures. The dynamics of reproductive conflicts in insect societies are directly connected to the colony kin structure, thus, the study of relatedness patterns is essential in order to understand the evolutionary resolution of these conflicts. We studied colony kin structure and mating frequencies in two closely related Neotropical ant species Pachycondyla inversa and Pachycondyla villosa. These represent interesting model systems because queens found new colonies cooperatively but, unlike many other ant species, they may still co-exist when the colony becomes mature (primary polygyny). By using five specific and highly variable microsatellite markers, we show that in both species queens usually mate with two or more males and that cofounding queens are always unrelated. Polygynous and polyandrous colonies are characterized by a high genetic diversity, with a mean relatedness coefficient among worker nestmates of 0.27 (+/- 0.03 SE) for P. inversa and 0.31 (+/- 0.05 SE) for P. villosa. However, relatedness among workers of the same matriline is high (0.60 +/- 0.03 in P. inversa, 0.62 +/- 0.08 in P. villosa) since males that mated with the same queen are on average closely related. Hence, we have found a new taxon in social Hymenoptera with high queen-mating frequencies and with intriguing mating and dispersal patterns of the sexuals.     

The determinants of queen size in a socially polymorphic ant

In social animals, body size can be shaped by multiple factors, such as direct genetic effects, maternal effects, or the social environment. In ants, the body size of queens correlates with the social structure of the colony: colonies headed by a single queen (monogyne) generally produce larger queens that are able to found colonies independently, whereas colonies headed by multiple queens (polygyne) tend to produce smaller queens that stay in their natal colony or disperse with workers. We performed a cross‐fostering experiment to investigate the proximate causes of queen size variation in the socially polymorphic ant Formica selysi. As expected if genetic or maternal effects influence queen size, eggs originating from monogyne colonies developed into larger queens than eggs collected from polygyne colonies, be they raised by monogyne or polygyne workers. In contrast, eggs sampled in monogyne colonies were smaller than eggs sampled in polygyne colonies. Hence, eggs from monogyne colonies are smaller but develop into larger queens than eggs from polygyne colonies, independently of the social structure of the workers caring for the brood. These results demonstrate that a genetic polymorphism or maternal effect transmitted to the eggs influences queen size, which probably affects the social structure of new colonies.     

Alternative genetic foundations for a key social polymorphism in fire ants

Little is known about the genetic foundations of colony social organization. One rare example in which a single major gene is implicated in the expression of alternative social organizations involves the presumed odorant-binding protein gene Gp-9 in fire ants. Specific amino acid substitutions in this gene invariably are associated with the expression of monogyny (single queen per colony) or polygyny (multiple queens per colony) in fire ant species of the Solenopsis richteri clade. These substitutions are hypothesized to alter the abilities of workers to recognize queens and thereby regulate their numbers in a colony. We examined whether these same substitutions underlie the monogyny/polygyny social polymorphism in the distantly related fire ant S. geminata. We found that Gp-9 coding region sequences are identical in the polygyne and monogyne forms of this species, disproving our hypothesis that one or a few specific amino acid replacements in the protein are necessary to induce transitions in social organization in fire ants. On the other hand, polygyne S. geminata differs genetically from the monogyne form in ways not mirrored in the two forms of S. invicta, a well-studied member of the S. richteri clade, supporting the conclusion that polygyny did not evolve via analogous routes in the two lineages. Specifically, polygyne S. geminata has lower genetic diversity and different gene frequencies than the monogyne form, suggesting that the polygyne form originated via a founder event from a local monogyne population. These comparative data suggest an alternative route to polygyny in S. geminata in which loss of allelic variation at genes encoding recognition cues has led to a breakdown in discrimination abilities and the consequent acceptance of multiple queens in colonies.     

Genetic evidence for landscape effects on dispersal in the army ant Eciton burchellii

Inhibited dispersal, leading to reduced gene flow, threatens populations with inbreeding depression and local extinction. Fragmentation may be especially detrimental to social insects because inhibited gene flow has important consequences for cooperation and competition within and among colonies. Army ants have winged males and permanently wingless queens; these traits imply male‐biased dispersal. However, army ant colonies are obligately nomadic and have the potential to traverse landscapes. Eciton burchellii, the most regularly nomadic army ant, is a forest interior species: colony raiding activities are limited in the absence of forest cover. To examine whether nomadism and landscape (forest clearing and elevation) affect population genetic structure in a montane E. burchellii population, we reconstructed queen and male genotypes from 25 colonies at seven polymorphic microsatellite loci. Pairwise genetic distances among individuals were compared to pairwise geographical and resistance distances using regressions with permutations, partial Mantel tests and random forests analyses. Although there was no significant spatial genetic structure in queens or males in montane forest, dispersal may be male‐biased. We found significant isolation by landscape resistance for queens based on land cover (forest clearing), but not on elevation. Summed colony emigrations over the lifetime of the queen may contribute to gene flow in this species and forest clearing impedes these movements and subsequent gene dispersal. Further forest cover removal may increasingly inhibit Eciton burchellii colony dispersal. We recommend maintaining habitat connectivity in tropical forests to promote population persistence for this keystone species.     

Gene flow is maintained by polyandry and male dispersal in the army ant <Emphasis Type="Italic">Eciton burchellii</Emphasis>

The combination of haplodiploidy, complementary sex determination and eusociality constrains the effective population size (N _e) of social Hymenoptera far more than in any other insect group. Additional limitations on N _e occur in army ants since they have wingless queens and colony fission, both of which are factors causing restricted maternal gene flow and high population viscosity. Therefore, winged army ant males gain a particular significance to ensure dispersal, facilitate gene flow and avoid inbreeding. Based on population genetic analyses with microsatellite markers, we studied a population of the Neotropical army ant Eciton burchellii, finding a high level of heterozygosity, weak population differentiation and no evidence for inbreeding. Moreover, by using sibship reconstruction analyses, we quantified the actual number of male contributing colonies represented in a queen’s mate sample, demonstrating that, through extreme multiple mating, the queens are able to sample the genes of males from up to ten different colonies, usually located within an approximate radius of 1 km. We finally correlated the individual mating success of each male contributing colony with the relative siring success of individual males and found a significant colony-dependent male fitness component. Our results imply that the dispersal and mating system of these army ants seem to enhance gene flow and minimise the deleterious effects associated with small effective population size. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Hierarchical Analysis of Genetic Structure in Native Fire Ant Populations: Results from Three Classes of Molecular Markers

We describe genetic structure at various scales in native populations of the fire ant Solenopsis invicta using two classes of nuclear markers, allozymes and microsatellites, and markers of the mitochondrial genome. Strong structure was found at the nest level in both the monogyne (single queen) and polygyne (multiple queen) social forms using allozymes. Weak but significant microgeographic structure was detected above the nest level in polygyne populations but not in monogyne populations using both classes of nuclear markers. Pronounced mitochondrial DNA (mtDNA) differentiation was evident also at this level in the polygyne form only. These microgeographic patterns are expected because polygyny in ants is associated with restricted local gene flow due mainly to limited vagility of queens. Weak but significant nuclear differentiation was detected between sympatric social forms, and strong mtDNA differentiation also was found at this level. Thus, queens of each form seem unable to establish themselves in nests of the alternate type, and some degree of assortative mating by form may exist as well. Strong differentiation was found between the two study regions using all three sets of markers. Phylogeographic analyses of the mtDNA suggest that recent limitations on gene flow rather than longstanding barriers to dispersal are responsible for this large-scale structure.     

Strict monandry in the ponerine army ant genus Simopelta suggests that colony size and complexity drive mating system evolution in social insects

Altruism in social insects has evolved between closely related full-siblings. It is therefore of considerable interest why some groups have secondarily evolved low within-colony relatedness, which in turn affects the relatedness incentives of within-colony cooperation and conflict. The highest queen mating frequencies, and therefore among the lowest degrees of colony relatedness, occur in Apis honeybees and army ants of the subfamilies Aenictinae, Ecitoninae, and Dorylinae, suggesting that common life history features such as reproduction by colony fission and male biased numerical sex-ratios have convergently shaped these mating systems. Here we show that ponerine army ants of the genus Simopelta, which are distantly related but similar in general biology to other army ants, have strictly monandrous queens. Preliminary data suggest that workers reproduce in queenright colonies, which is in sharp contrast to other army ants. We hypothesize that differences in mature colony size and social complexity may explain these striking discrepancies.     

Mating frequency of ant queens with alternative dispersal strategies, as revealed by microsatellite analysis of sperm

In social Hymenoptera (ants, bees, and wasps), the number of males that mate with the same queen affects social and genetic organization of the colony. However, the selective forces leading to single mating in certain conditions and multiple mating in others remain enigmatic. In this study, I investigated whether queens of the wood ant Formica paralugubris adopting different dispersal strategies varied in their mating frequency (the number of males with whom they mated). The frequency of multiple mating was determined by using microsatellite markers to genotype the sperm stored in the spermatheca of queens, and the validity of this method was confirmed by analysing mother–offspring combinations obtained from experimental single-queen colonies. Dispersing queens, which may found new colonies, did not mate with more males than queens that stayed within polygynous colonies, where the presence of numerous reproductive individuals ensured a high level of genetic diversity. Hence, this study provides no support to the hypotheses that multiple mating is beneficial because it increases genetic variability within colonies. Most of the F. paralugubris queens mated with a single male, whatever their dispersal strategy and life history. Moreover, multiple mating had little effect on colony genetic structure: the effective mating frequency was 1.11 when calculated from within-brood relatedness, and 1.13 when calculated from the number of mates detected in the sperm. Hence, occasional multiple mating by F. paralugubris queens may have no adaptive significance.     

Polygynous supercolonies of the acacia‐ant Pseudomyrmex peperi,an inferior colony founder

In ant–plant protection mutualisms, plants provide nesting space and nutrition to defending ants. Several plant–ants are polygynous. Possessing more than one queen per colony can reduce nestmate relatedness and consequently the inclusive fitness of workers. Here, we investigated the colony structure of the obligate acacia‐ant Pseudomyrmex peperi, which competes for nesting space with several congeneric and sympatric species. Pseudomyrmex peperi had a lower colony founding success than its congeners and thus, appears to be competitively inferior during the early stages of colony development. Aggression assays showed that P. peperi establishes distinct, but highly polygynous supercolonies, which can inhabit large clusters of host trees. Analysing queens, workers, males and virgin queens from two supercolonies with eight polymorphic microsatellite markers revealed a maximum of three alleles per locus within a colony and, thus, high relatedness among nestmates. Colonies had probably been founded by one singly mated queen and supercolonies resulted from intranidal mating among colony‐derived males and daughter queens. This strategy allows colonies to grow by budding and to occupy individual plant clusters for time spans that are longer than an individual queen’s life. Ancestral states reconstruction indicated that polygyny represents the derived state within obligate acacia‐ants. We suggest that the extreme polygyny of Pseudomyrmex peperi, which is achieved by intranidal mating and thereby maintains high nestmate relatedness, might play an important role for species coexistence in a dynamic and competitive habitat.     

Effects of social organization and elevation on spatial genetic structure in a montane ant

Studying patterns of population structure across the landscape sheds light on dispersal and demographic processes, which helps to inform conservation decisions. Here, we study how social organization and landscape factors affect spatial patterns of genetic differentiation in an ant species living in mountainous regions. Using genome‐wide SNP markers, we assess population structure in the Alpine silver ant, Formica selysi. This species has two social forms controlled by a supergene. The monogyne form has one queen per colony, while the polygyne form has multiple queens per colony. The two social forms co‐occur in the same populations. For both social forms, we found a strong pattern of isolation‐by‐distance across the Alps. Within regions, genetic differentiation between populations was weaker for the monogyne form than for the polygyne form. We suggest that this pattern is due to higher dispersal and effective population sizes in the monogyne form. In addition, we found stronger isolation‐by‐distance and lower genetic diversity in high elevation populations, compared to lowland populations, suggesting that gene flow between F. selysi populations in the Alps occurs mostly through riparian corridors along lowland valleys. Overall, this survey highlights the need to consider intraspecific polymorphisms when assessing population connectivity and calls for special attention to the conservation of lowland habitats in mountain regions.     

Selective male mortality in the red imported fire ant, Solenopsis invicta

Males in polygyne populations of Solenopsis invicta are primarily sterile diploids and thought to not express the Gp-9 gene coding for a pheromone-binding protein affecting complex social behavior. We examined an aspect of the breeding system hitherto not considered--male Gp-9 genotypes in relation to sperm stored in queens. Four sites with varying frequencies of sympatric monogyne and polygyne colonies were sampled, including sexuals, workers, and broods from four colonies. Most queens were heterozygotes storing B sperm. Although predicted to be common, only 14 of 504 males were B or BB genotypes, suggesting strong selection. Increased frequency of polygyne colonies at each site paralleled increases in queens with b sperm (1.9-32.8%) and of noninseminated queens. The presence of both B and b sperm in 1.9-18.9% of queens, genotype profiles of colonies, and genotypes of offspring from individual queens suggest some frequency of multiple mating. The bb genotype, rather than an obligate, developmental lethal, was present in some queens and common in alates, workers, and brood. Selective mortality of sexuals may affect multiple aspects of the breeding system, including female-mediated dispersal, mating success, and gene flow.     

Facultative polygyny and habitat succession in boreal ants

High dispersal risks of ant queens make staying in the natal patch more attractive than long range dispersal. These alternative strategies and the mode of colony founding determine the average number of queens in the population. Increasing competition and queen predation make independent colony founding increasingly difficult and the only option for new queens to reproduce in the habitat patch may be to enter an existing colony. The effect of nest-site availability to the number of queens was studied in successional spruce-dominated taiga forests in facultatively polygynous ants Myrmica ruginodis, M. sulcinodis, Leptothorax acervorum, Formica sanguinea and F. truncorum. Decreasing relatedness among worker nestmates supports an association between increasing habitat age and polygyny to some extent. M. sulcinodis and L. acervorum persist in this type of taiga only for a relatively short period. Relatedness varied only slightly among populations, but lower relatedness estimated in other studies suggested higher levels of polygyny in older populations. In M. ruginodis there was more variation in relatedness and it was possibly connected to the relative proportions of the two social forms of the species. In F. sanguinea and F. truncorum the decrease in relatedness with increasing age of the habitat was clearest. Other factors favouring limited dispersal and acceptance of new queens in the colonies are, however, hard to separate.     

Mating Triggers Queen Elimination by Workers of Japanese Harvester Ant (<Emphasis Type="Italic">Messor aciculatus</Emphasis>)

During reproduction, ant colonies produce winged queens. These new queens usually leave the nest to mate and can then establish a new nest. If the new nest is close to an existing colony, it will be in competition with the existing colony. Therefore, workers will kill any mated queens they find outside the colony during the reproductive season. In this study, factors that might determine whether workers eliminate queens were investigated. Mating status (mated or unmated), colony origin (same or different to tested workers) and mating partners (inbred or outbred) of the queens of Japanese harvester ants (Messor aciculatus) were manipulated and the workers’ behavior towards the queens was observed. Mated queens were always attacked by workers, though this was not affected by either colony origin or mating partners. These results suggest that mating status triggers elimination of queens by workers, and that the colony origin and mating partner are unlikely to be important roles in elimination of queens.     

Habitat structure,dispersal strategies and queen number in two boreal Leptothorax ants

In two nearctic ants, Leptothorax canadensis and Leptothorax sp. A, young queens may either found their own nest solitarily after mating or seek adoption into an established colony. Whether a queen disperses or not is associated with genetically determined queen morphology in Leptothorax sp. A. Whereas a majority of winged queens attempt solitary colony founding after mating, most wingless, intermorphic queens return to their maternal nests and new colonies are founded by budding after hibernation. The latter strategy appears to be correlated with patchy, isolated habitats, whereas in extended boreal forests dispersal on the wing is probably more common. Alternative dispersal strategies strongly affect the average number of queens per colony and seasonal fluctuations of colony structure.