Seasonal cycle of colony structure in the Ponerine ant Pachycondyla chinensis in western Japan (Hymenoptera, Formicidae)

Investigation of reproductive strategies of ants in the subfamily Ponerinae is important for understanding of the evolution of social structure and of the significance of caste dimorphisms. The biology of species with mated and egg-laying workers (gamergates) has been studied for many species, however, little attention has been paid to species that reproduce via alate queens only. We investigated the seasonal cycle of changes in the colony structure of Pachycondyla chinensis reproduced by alate queens in western Japan, and found the following novel biological characteristics of this species. P. chinensis showed a remarkable caste dimorphism in ovariole numbers: workers had no ovaries while queens had 18 to 36 ovarioles in their ovaries. The nesting system seemed to be polydomous: 266 of 400 nests collected were queenless. The number of queenless nests increased during the reproductive season. Among the 134 queenright nests, 38 had several mated-queens without significant differences in ovary activation and the remaining 96 nests were monogynous. During winter to early spring, most nests were polygynous. After alate production, most of the old queens seemed to die or be expelled and replaced by new queens. Virgin dealated queens were often found and they seemed to have laid eggs. Received 3 August 2007; revised 19 December 2007; accepted 20 December 2007     

Breeding behaviour, relatedness and sex-investment ratios inLeptothorax tuberum Fabricius

The ant speciesLeptothorax tuberum was shown to be predominantly monogynous. Queens usually mate once only but some nests may have a multiply-mated queen or are partially or serially polygynous. As expected from these results, within nest relatedness between workers and between workers and alate queens was found to be high. Almost fifty percent of nests had no nest queen which may indicate high queen mortality, queens leaving to found new nests or nest fragmentation. Observed female investment frequencies (IF _o ) were not significantly different from those expected on the basis of worker control of sexual production and the relatedness estimates of workers to alate queens and workers to males calculated from isozyme data (IF _E ). These values were not consistent with queen control. There was no evidence for lower IF _o s in queenless nests nor for higher IF _o s in larger nests classified by worker number. When nests were classified by sexual productivity, however, there was a strong rank correlation between productivity and female bias. This is the first study of an ant species to test observed IF against expected IF calculated without inferring between caste relatedness from worker data or pooling of data from different castes.     

Dimorphic ergatoid males and their reproductive behavior in the ponerine antHypoponera bondroiti

Summary We studied the reproductive behavior of the ponerine antHypoponera bondroiti from Okinawa, Japan. This species has dimorphic wingless ergatoid males (major and minor), dimorphic reproductive females (alate queens and wingless reproductive intercastes), and workers. Workers have neither ovarioles nor spermatheca. Major ergatoid males are the largest colony members. Two major males fought one another in the nest until one disappeared, leaving the other to occupy the nest chambers where queens emerge and mate. Minor ergatoid males also fought one another, although they seemed to be less pugnacious, resulting in occasional cohabitation of multiple minor males in the same nest chamber. Major males never attacked minor ones, allowing them to coexist in the same nest chamber. Minor males seemed to mimic females. Both major and minor males mated with both alate queens and intercastes within the nest. After mating, some alate queens shed their wings and remained in the nest, while the others left the nest for dispersal in the laboratory. Intercastes remained in the nest.     

Competition over workers: fertility signalling in wingless queens of <Emphasis Type="Italic">Hypoponera opacior</Emphasis>

The ant Hypoponera opacior exhibits alternative reproductive morphs associated with distinct sexual behaviours. Wingless, worker-like gynes mate in fall in their mother nest. Subsequently, these polygynous nests divide and wingless queens disperse on foot accompanied by workers, which help during nest foundation. Local resource competition amongst wingless queens was indicated by male-biased sex ratios. Queens compete over access to workers, and this competition manifests in dominance interactions. We colour-marked queens and workers to study this behaviour. Marked queens were treated aggressively and frequently killed by nestmates, whilst similarly labelled workers were readopted. We hypothesise that mated wingless queens—although they resemble workers in external morphology—differ chemically and that fertile queens with divergent odours are not tolerated. Thereby, sterile H. opacior workers avoid the risk to raise offsprings of unrelated queens. However, when nests split, workers should base their decision which queen to follow not only on relatedness, but also on queen fertility. Cuticular hydrocarbon profiles of queens could serve as an honest signal for workers. Indeed, gas chromatography showed that hydrocarbon composition differs between workers and mated queens and is linked to ovary development in queens. Workers and unmated queens exhibit complex profiles of short-chained hydrocarbons, whilst fertile queens carry few, long-chained alkenes and branched alkanes. Furthermore, ovary length was correlated to the relative amounts of characteristic hydrocarbons of queens. The chemical profile was also associated with eye size—an unalterable trait in adult insects—suggesting that larval nutrition and consequently body size affects adult ovary development.     

Thelytokous parthenogenesis in the exotic dacetine ant Strumigenys rogeri (Hymenoptera: Formicidae) in Taiwan

Only recently has it become clear that several species of eusocial hymenopterans regularly reproduce by thelytokous parthenogenesis, that is, the production of diploid female offspring by unmated females. This phenomenon suggests that parthenogenetic reproduction might be advantageous to organisms under certain environmental conditions. Here the occurrence of asexual reproduction is reported for the first time in the dacetine ant, Strumigenys rogeri, at least for the focal populations in Taiwan. Virgin queens of S. rogeri maintained with several workers produced both workers and young queens from unfertilized eggs under laboratory conditions in as short as 39 days, whereas workers were strictly sterile as no spermatheca was discovered after dissection. Combined with additional evidence (i.e. absence of males in field colonies), queen thelytoky is confirmed. Such a reproductive mode and short development time may jointly help explain the success of this tramp ant species in Taiwan and elsewhere.     

Proximate Determinants of Reproductive Skew in Polygyne Colonies of the Ant Formica fusca

Understanding the determinants of reproductive skew (the partitioning of reproduction among co‐breeding individuals) is one of the major questions in social evolution. In ants, multiple‐queen nests are common and reproductive skew among queens has been shown to vary tremendously both within and between species. Proximate determinants of skew may be related to both queen and worker behaviour. Queens may attempt to change their reproductive share through dominance interactions, egg eating and by changing individual fecundity. Conversely, workers are in a position to regulate the reproductive output of queens when rearing the brood. This paper investigates queen behaviour at the onset of egg laying and the effect of queen fecundity and worker behaviour on brood development and reproductive shares of multiple queens in the ant Formica fusca. The study was conducted in two‐queen laboratory colonies where the queens produced only worker offspring. The results show that in this species reproductive apportionment among queens is not based on dominance behaviour and aggression, but rather on differences in queen fecundity. We also show that, although the queen fecundity at the onset of brood rearing is a good indicator of her final reproductive output, changes in brood composition occur during brood development. Our results highlight the importance of queen fecundity as a major determinant of her reproductive success. They furthermore suggest that in highly derived polygyne species, such as the Formica ants, direct interactions as a means for gaining reproductive dominance have lost their importance.     

Evolution of wingless reproductives in ants: weakly specialized ergatoid queen instead of gamergates in Platythyrea conradti

Platythyrea conradti is the only species in this genus with ergatoid (= permanently wingless) queens. Colonies lack gamergates (unlike other species in this genus), yet aggressive interactions among queen and workers define a hierarchy. A single fertile queen has the top rank and highranking workers do not lay eggs, except when the queen dies. Colonial reproduction by both alate queens (independent foundation) and gamergates (fission) seems the ancestral state in Platythyrea. Independent foundation can be selected against in some species, causing the loss of alate queens for economic reasons. Thus gamergates become the only reproductives, except in P. conradti in which queens became ergatoid. Gamergates and ergatoid queens are two mutually redundant reproductive phenotypes that allow colonial reproduction by fission. Received 1 July 2005; revised 10 November 2005; accepted 16 November 2005.     

The caste system of the dolichoderine antTechnomyrmex albipes (Hymenoptera: Formicidae): morphological description of queens,workers and reproductively active intercastes

Females ofTechnomyrmex albipes consist of winged queens, intercastes and workers. In established colonies, reproduction is performed by many intercastes (wingless females which have intermediate phenotypes between queen and worker characters). Dissection and morphological examination revealed that intercastes had a spermatheca, but workers did not. Intercastes can be divided into three classes: major intercastes with three ocelli, medium intercastes with one ocellus, and minor intercases without ocelli. Workers have no ocelli. The thoracic structure of intercastes gradually becomes more complex from minors to majors. The body size of intercastes gradually increases from minors to majors, and so does the number of overioles. The body size distributions of minor intercastes and workers overlap considerably, but the distributions of ovariole numbers overlap less. Winged queens had distinctly larger body sizes, more ovarioles and larger spermathecae than intercastes. Most intercastes were inseminated with developed ovaries and appeared to reproduce. The caste system and reproductive division of labour inT. albipes is compared to those of ant species in which permanently wingless females reproduce.     

Stay or drift? Queen acceptance in the ant <Emphasis Type="Italic">Formica paralugubris</Emphasis>

The acceptance of new queens in ant colonies has profound effects on colony kin structure and inclusive fitness of workers. Therefore, it is important to study the recognition and discrimination behaviour of workers towards reproductive individuals entering established colonies. We examined the acceptance rate of queens in populations of the highly polygynous ant F. paralugubris, where the genetic differentiation among nests and discrimination ability among workers suggest that workers might reject foreign queens. We experimentally introduced young queens in their natal nest and in foreign nests. Surprisingly, the survival rate of mated queens did not differ significantly when introduced in a foreign male-producing nest, a foreign female-producing nest, or the natal nest. Moreover, the survival of virgin queens in their natal nest was twice the one of mated queens, suggesting that mating status plays an important role for acceptance. The results indicate that other factors than queen discrimination by workers are implicated in the limited longdistance gene flow between nests in these populations. Received 8 April 2008; revised 16 June 2008; accepted 1 July 2008.     

Similarity in venom alkaloid chemistry of alate queens of imported fire ants: implication for hybridization between Solenopsis richteri and S. invicta in the Southern United States

Both cis- and trans-2-methyl-6-undecylpiperidines, MC11P, have been previously reported as the major components of the venom of alate queens of the imported fire ants, Solenopsis richteri (black) and S. invicta (red). To identify the minor components of venom alkaloids from alate queens and compare the venom alkaloid chemistry of alate queen of their hybrid (S. richteri×S. invicta) with that of the two parental fire ant species (S. richteri and S. invicta), silica-gel short-column chromatography was utilized for separating cis-stereoisomers of venom alkaloids from trans-stereoisomers. GC/MS Analyses of venom-alkaloid chemistry of alate queens demonstrated that fewer alkaloid peaks were detected in the chromatograms of the alate queens compared to those of workers. Three new compounds, 7, 12, and 13, were detected as minor components in the venom of alate queens of all three fire ant species. Alate queens of hybrid fire ants showed cis- and trans-alkaloid patterns similar to those of the parental species. Similarity in venom-alkaloid chemistry of alate queens of S. richteri and S. invicta, and their hybrid may indicate their reproductive compatibility in the hybrid zone in southern United States, where all three species occur sympatrically.     

Unusual Behavior of Polygyne Fire Ant Queens on Nuptial Flights

This study reports previously undescribed behavior of fire ant queens (Solenopsis invicta) on their nuptial flights. We captured large numbers of alate (winged) queens flying at low altitudes in dense swarms that were virtually devoid of males. We assayed the genotypes of these alate queens at the locus Gp-9, which exhibits strong genotype frequency differences between monogyne (single-queen) and polygyne (multiple-queen) populations, and found that almost all of these low-flying queens originated from polygyne colonies. Comparisons of mtDNA haplotype distributions of these queens to those of alates leaving polygyne nests suggest that the flying queens had not dispersed more than a few hundred meters. Moreover, the proportion of flying queens that were mated did not differ significantly from the proportion of reproductive queens that were mated within the same sites. Thus the flight behavior appears to occur subsequent to mating. We suggest that the flying queens are sampling the local environment in order to select a suitable landing site. Such a site would contain established polygyne nests into which the queens may be adopted as new reproductives.     

Ovarian activity correlates with extreme changes in cuticular hydrocarbon profile in the highly polygynous ant, Linepithema humile

In social insects, cuticular hydrocarbons of adults may vary with ovarian activity. Such variations are suggested to function in the regulation of reproduction within colonies. The modification of the CH profile with ovarian activity is usually interpreted as a signal of fertility causing workers to refrain from reproducing in the presence of the queen. We examined the effect of ovarian activity on the CH profiles in the Argentine ant Linepithema humile, a species where workers lack ovaries and are completely sterile. Our data show considerable differences in the CH profiles between fertile and infertile individuals. These differences are mainly qualitative. The CH profile strongly changes at the start of egg laying, both in mated and unmated queens. These results show that variation in cuticular hydrocarbons with ovarian activity in ants is not restricted to species with worker reproduction. We propose that in the Argentine ant, the cuticular hydrocarbons of laying queens correspond to a signal of fertility involved in the regulation of various aspects of reproduction, such as the rearing of new sexuals.     

Social Control over the Survival and Selection of Winged Virgin Queens in an Ant without Nuptial Flight: Iridomyrmex humilis

Laboratory and field experiments show that the number of winged virgin queens of the Argentine ant Iridomyrmex humilis allowed to remain alive in the nest is under social control. When reared in their natal nest, survival of virgin queens depends on the presence of males. In the presence of mature alate males or even male pupae only 20% of the gynes are executed before they are 4 days old. By contrast, if pupal and adult males are absent, about 55% of the gynes are executed. The workers tend to kill the lightest ones. Although the Argentine ant is a unicolonial species exhibiting complete acceptance of foreign conspecific workers and mated queens, regardless of whether the recipient nest is queenright or queenless, the acceptance of gynes is variable: they are executed by foreign queenless workers but widely accepted in queenright nests. These results are discussed with regard to possible existence of queen and gyne pheromones allowing recognition and to the lack of nuptial flight in this species.     

Colony structure and reproduction in the thelytokous parthenogenetic ant Platythyrea punctata (F. Smith) (Hymenoptera, Formicidae)

Summary: An important evolutionary characteristic of the formicine subfamily Ponerinae is the occurrence of various alternative reproductive tactics within single species. In Platythyrea punctata Smith, 1858, queens, gamergates and parthenogenetic workers co-occur in the same species. Morphological queens, both alate and dealate, were present in only 29 percent of the colonies collected in Florida, but absent from colonies collected in Barbados and Puerto Rico. One of the six queens which were dissected (three alate and three dealate) was found to be inseminated but not fertile. Instead, in most queenless colonies, a single uninseminated worker monopolized reproduction by means of thelytokous parthenogenesis, i.e., it produced female offspring from unfertilized eggs. A single mated, reproductive worker (gamergate) was found dominating reproduction in the presence of an inseminated alate queen in one of the Florida colonies. Thelytokous parthenogenesis was examined in artificial groups of virgin laboratory-reared workers, where one worker typically monopolized reproduction despite the presence of several individuals with elongated ovaries. In 16 colonies collected in Florida, a total of 66 individuals differed morphologically from queens and workers. Their thorax morphology varied from a worker-like to an almost queen-like structure. We refer to these individuals as "intercastes" (sensu Peeters, 1991a). The remarkable complexity of reproductive strategies renders P. punctata unique within ants.     

Repeated evolution of queen parthenogenesis and social hybridogenesis in Cataglyphis desert ants

Over the last decade, genetic studies on social insects have revealed a remarkable diversity of unusual reproductive strategies, such as male clonality, female clonality, and social hybridogenesis. In this context, Cataglyphis desert ants are useful models because of their unique reproductive systems. In several species, queens conditionally use sexual reproduction and parthenogenesis to produce sterile workers and reproductive queens, respectively. In social hybridogenesis, two distinct genetic lineages coexist within a population, and workers result from mating between partners of different lineages; in contrast, queens and males are both produced asexually by parthenogenesis. Consequently, nonreproductive workers are all interlineage hybrids, whereas reproductives are all pure lineage individuals. Here, we characterized the reproductive systems of 11 species to investigate the distribution of the conditional use of sex and social hybridogenesis in Cataglyphis. We identified one new case in which sexual reproduction was conditionally used in the absence of dependent‐lineage reproduction. We also discovered five new instances of social hybridogenesis. Based on our phylogenetic analyses, we inferred that both the conditional use of sex and social hybridogenesis independently evolved multiple times in the genus Cataglyphis.     

Genome-wide analysis reveals differences in brain gene expression patterns associated with caste and reproductive status in honey bees (Apis mellifera)

A key characteristic of eusocial species is reproductive division of labour. Honey bee colonies typically have a single reproductive queen and thousands of sterile workers. Adult queens differ dramatically from workers in anatomy, physiology, behaviour and lifespan. Young female workers can activate their ovaries and initiate egg laying; these 'reproductive' workers differ from sterile workers in anatomy, physiology, and behaviour. These differences, however, are on a much smaller scale than those observed between the queen and worker castes. Here, we use microarrays to monitor expression patterns of several thousand genes in the brains of same-aged virgin queens, sterile workers, and reproductive workers. We found large differences in expression between queens and both worker groups (~2000 genes), and much smaller differences between sterile and reproductive workers (221 genes). The expression patterns of these 221 genes in reproductive workers are more queen-like, and may represent a core group of genes associated with reproductive physiology. Furthermore, queens and reproductive workers preferentially up-regulate genes associated with the nurse bee behavioural state, which supports the hypothesis of an evolutionary link between worker division of labour and molecular pathways related to reproduction. Finally, several functional groups of genes associated with longevity in other species are significantly up-regulated in queens. Identifying the genes that underlie the differences between queens, sterile workers, and reproductive workers will allow us to begin to characterize the molecular mechanisms underlying the evolution of social behaviour and large-scale remodelling of gene networks associated with polyphenisms.     

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.     

Genetic determination of female castes in a hybridogenetic desert ant

In most social insects, the brood is totipotent and environmental factors determine whether a female egg will develop into a reproductive queen or a functionally sterile worker. However, genetic factors have been shown to affect the female's caste fate in a few ant species. The desert ant Cataglyphis hispanica reproduces by social hybridogenesis. All populations are characterized by the coexistence of two distinct genetic lineages. Queens are almost always found mated with a male of the alternate lineage than their own. Workers develop from hybrid crosses between the genetic lineages, whereas daughter queens are produced asexually via parthenogenesis. Here, we show that the association between genotype and caste in this species is maintained by a ‘hard‐wired’ genetic caste determination system, whereby nonhybrid genomes have lost the ability to develop as workers. Genetic analyses reveal that, in a rare population with multiple‐queen colonies, a significant proportion of nestmate queens are mated with males of their own lineage. These queens fail to produce worker offspring; they produce only purebred daughter queens by sexual reproduction. We discuss how the production of reproductive queens through sexual, intralineage crosses may favour the stability of social hybridogenesis in this species.     

Behavioral property of unusual large workers in the ant,<Emphasis Type="Italic"> Pristomyrmex pungens</Emphasis>

We surveyed the behavioral properties of unusual large workers in the parthenogenetic queenless ant, Pristomyrmex pungens. The majority of colonies mostly consist of monomorphic workers, which have two ovarioles (small workers). Large workers with four ovarioles are occasionally found in some colonies. We compared the behavior of small and large workers. Large workers performed brood care less frequently than small workers and almost never foraged. We set out three hypotheses of the adaptive significance of large workers: they are (1) reproductive castes, that is, functionally defined queens, (2) major workers, such as soldiers, and (3) of a socially parasitic lineage. Electronic Publication     

Queen adoption in the polygynous and polydomous ant, Leptothorax curvispinosus

Alate female reproductives of the facultatively polygynous andpolydomous ant, Lepiothorax curuispinosus, were reared fromfield-collected nests, mated, and introduced into either theirparental nests or alien conspecific nests. The 41 queens introducedinto alien nests were usually attacked and rejected (97. 6%),but one queen was accepted after initial aggression. The 27queens introduced into their parental nests received a variableresponse. Some were accepted without any apparent aggression(59. 3%), but others were strongly attacked and rejected (40.7%). Sequential introductions of up to four queens into particularparental nests indicated that nests consistently either acceptor reject their mated offspring. The presence or absence ofresident queens in parental nests had no apparent influenceon the acceptance of offspring queens. Nests that accepted queenshad significantly fewer workers than those that did not, butthis slight difference is unlikely to explain these dichotomousresults and could be spurious. Dissections of the introducedqueens revealed that 79. 0% were inseminated and 98. 3% haddeveloping, yolked eggs in their ovarioles, but these variableshad no apparent effect on acceptability. Similarly, the sizeof the introduced queens and the time that elapsed between matingand introduction had no apparent effect. The consistent responseof parental nests in either accepting or rejecting their matedoffspring indicates a mechanism of queen number regulation inthis species that involves characteristics of the colony ornest rather than variability among offspring queens. This mechanismcould be responsible for maintaining relatively low numbersof queens and high genetic relatedness in colonies (or individualnests) while promoting flexibility in colony reproduction bycolony fission ("budding") and the dispersal of young queens.This mechanism could also involve an important conflict of interestbetween parental colonies and their mated offspring and mightcontribute to the evolution of socially parasitic colony foundationstrategies. Acceptance of mated offspring by their parentalcolonies might only occur during certain periods in colony development,depend on the level of genetic diversity within the colony (ornest), reflect the condition of the colony, nest, queen(s),brood, or local habitat, or result from a genetic polymorphism.