Genetic and environmental effects on morphological traits of social phenotypes in wasps.

Many species exhibit distinct phenotypic classes, such as sexes in dioecious species or castes in social species. The evolution of these classes is affected by the genetic architecture governing traits shared between phenotypes. However, estimates of the genetic and environmental factors contributing to phenotypic variation in distinct classes have rarely been examined. We studied the genetic architecture underlying morphological traits in phenotypic classes in the social wasp Vespula maculifrons. Our data revealed patriline effects on a few traits, indicating weak genetic influences on caste phenotypic variation. Interestingly, traits exhibited higher heritability in queens than workers. This result suggests that genetic variation has a stronger influence on trait variation in the queen caste than the worker caste, which is unexpected because queens typically experience direct selection. Moreover, estimates of heritability for traits were correlated between the castes, indicating that variability in trait size was governed by similar genetic architecture in the two castes. However, we failed to find evidence for a significant relationship between caste dimorphism and caste correlation, as would be expected if trait evolution was constrained by intralocus genetic conflict. Our analyses also uncovered variation in the allometric relationships for traits. These analyses suggested that worker traits were proportionally smaller than queen traits for most traits examined. Overall, our data provide evidence for a strong environmental and moderate genetic basis of trait variation among castes. Moreover, our results suggest that selection previously operated on caste phenotype in this species, and phenotypic variation is now governed primarily by environmental differences.

Aphid secondary symbionts do not affect prey attractiveness to two species of predatory lady beetles.

Heritable symbionts have been found to mediate interactions between host species and their natural enemies in a variety of organisms. Aphids, their facultative symbionts, and their potential fitness effects have been particularly well-studied. For example, the aphid facultative symbiont Regiella can protect its host from infection from a fungal pathogen, and aphids with Hamiltonella are less likely to be parasitized by parasitic wasps. Recent work has also found there to be negative fitness effects for the larvae of two species of aphidophagous lady beetles that consumed aphids with facultative symbionts. In both species, larvae that consumed aphids with secondary symbionts were significantly less likely to survive to adulthood. In this study we tested whether adult Harmonia axyridis and Hippodamia convergens lady beetles avoided aphids with symbionts in a series of choice experiments. Adults of both lady beetle species were as likely to choose aphids with symbionts as those without, despite the potential negative fitness effects associated with consuming aphids with facultative symbionts. This may suggest that under natural conditions aphid secondary symbionts are not a significant source of selection for predatory lady beetles.

Evidence of indirect symbiont conferred protection against the predatory lady beetle Harmonia axyridis in the pea aphid.

BACKGROUND: Defensive symbionts can provide significant fitness advantages to their hosts. Facultative symbionts can protect several species of aphid from fungal pathogens, heat shock, and parasitism by parasitoid wasps. Previous work found that two of these facultative symbionts can also indirectly protect pea aphids from predation by the lady beetle Hippocampus convergens. When aphids reproduce asexually, there is extremely high relatedness among aphid clone-mates and often very limited dispersal. Under these conditions, symbionts may indirectly protect aphid clone-mates from predation by negatively affecting the survival of a predator after the consumption of aphids harboring the same vertically transmitted facultative symbionts. In this study, we wanted to determine whether this indirect protection extended to another lady beetle species, Harmonia axyridis. RESULTS: We fed Ha. axyridis larvae aphids from one of four aphid sub-clonal symbiont lines which all originated from the same naturally symbiont free clonal aphid lineage. Three of the sub-clonal lines harbor different facultative symbionts that were introduced to the lines via microinjection. Therefore these sub-clonal lineages vary primarily in their symbiont composition, not their genetic background. We found that aphid facultative symbionts affected larval survival as well as pupal survival in their predator Ha. axyridis. Additionally, Ha. axyridis larvae fed aphids with the Regiella symbiont had significantly longer larval developmental times than beetle larvae fed other aphids, and females fed aphids with the Regiella symbiont as larvae weighed less as adults. These fitness effects were different from those previously found in another aphid predator Hi. convergens suggesting that the fitness effects may not be the same in different aphid predators. CONCLUSIONS: Overall, our findings suggest that some aphid symbionts may indirectly benefit their clonal aphid hosts by negatively impacting the development and survival of a lady beetle aphid predator Ha. axyridis. By directly affecting the survival of predatory lady beetles, aphid facultative symbionts may increase the survival of their clone-mates that are clustered nearby and have significant impacts across multiple trophic levels. We have now found evidence for multiple aphid facultative symbionts negatively impacting the survival of a second species of aphid predatory lady beetle. These same symbionts also protect their hosts from parasitism and fungal infections, though these fitness effects seem to depend on the aphid species, predator or parasitoid species, and symbiont type. This work further demonstrates that beneficial mutualisms depend upon complex interactions between a variety of players and should be studied in multiple ecologically relevant contexts.

Aphid facultative symbionts reduce survival of the predatory lady beetle Hippodamia convergens.

BACKGROUND: Non-essential facultative endosymbionts can provide their hosts with protection from parasites, pathogens, and predators. For example, two facultative bacterial symbionts of the pea aphid (Acyrthosiphon pisum), Serratia symbiotica and Hamiltonella defensa, protect their hosts from parasitism by two species of parasitoid wasp. Previous studies have not explored whether facultative symbionts also play a defensive role against predation in this system. We tested whether feeding on aphids harboring different facultative symbionts affected the fitness of an aphid predator, the lady beetle Hippodamia convergens. RESULTS: While these aphid faculative symbionts did not deter lady beetle feeding, they did decrease survival of lady beetle larvae. Lady beetle larvae fed a diet of aphids with facultative symbionts had significantly reduced survival from egg hatching to pupation and therefore had reduced survival to adult emergence. Additionally, lady beetle adults fed aphids with facultative symbionts were significantly heavier than those fed facultative symbiont-free aphids, though development time was not significantly different. CONCLUSIONS: Aphids reproduce clonally and are often found in large groups. Thus, aphid symbionts, by reducing the fitness of the aphid predator H. convergens, may indirectly defend their hosts' clonal descendants against predation. These findings highlight the often far-reaching effects that symbionts can have in ecological systems.

Effects of size, shape, genotype, and mating status on queen overwintering survival in the social wasp Vespula maculifrons.

Many insects survive cold winter temperatures by entering into a transient, dormant state. Social wasp queens in the genus Vespula undergo such a state of physiological dormancy known as diapause to survive winter conditions. We experimentally investigated the effects of size, shape, genotype, and matedness on the overwintering survival of Vespula maculifrons Buysson (Hymenoptera: Vespidae) queens in two study years. Queens first were mated in the laboratory and then subjected to overwintering conditions. The sizes of several morphological traits of each queen were measured, and the genotype of each queen was determined at polymorphic microsatellite loci allowing queens to be assigned to distinct patrilines. We found that overall large body size resulted in high overwintering survival for queens in one study year. In addition, queen thinness was positively and significantly associated with overwintering survival in both years. In contrast, we found no evidence for an effect of patriline on overwintering survival, which is consistent with the idea that natural selection removes genetic variation associated with traits directly linked to fitness. We also found no effect of queen matedness on overwintering survival, as expected under models suggesting that mating should not adversely affect female viability in social hymenopteran insects. Overall, our study demonstrates that some aspects of body size and shape are significantly associated with overwintering survival in V. maculifrons queens. However, variation in survival arising from genotypic differences or mating effects is limited due, in part, to the way selection operates in social insect species.

Horizontally transferred fungal carotenoid genes in the two-spotted spider mite Tetranychus urticae.

Carotenoids are organic pigments commonly synthesized by plants, algae and some micro-organisms. Through absorption of light energy, carotenoids facilitate photosynthesis and provide protection against photo-oxidation. While it was presumed that all carotenoids in animals were sequestered from their diets, aphids were recently shown to harbour genomic copies of functional carotenoid biosynthesis genes that were acquired via horizontal gene transfer from fungi. Our search of available animal transcripts revealed the presence of two related genes in the two-spotted spider mite Tetranychus urticae. Phylogenetic analyses suggest that the T. urticae genes were transferred from fungi into the spider mite genome, probably in a similar manner as recently suggested for aphids. The genes are expressed in both green and red morphs, with red morphs exhibiting higher levels of gene expression. Additionally, there appear to be changes in the expression of these genes during diapause. As carotenoids are associated with diapause induction in these animals, our results add to recent findings highlighting the importance of eukaryotic horizontal gene transfer in the ecology and evolution of higher animals.

Resource allocation in a social wasp: effects of breeding system and life cycle on reproductive decisions.

Organisms must make important decisions on how to allocate resources to reproduction. We investigated allocation decisions in the social wasp Vespula maculifrons to understand how social insects make reproductive choices. We first determined how annual colonies apportioned resources to growth and reproduction by analysing developing brood. In contrast to expectations, colonies invested in both growth (workers) and reproduction (males) simultaneously. In addition, colonies showed evidence of producing males in pulses and reversing their reproductive choices by decreasing investment in males late in the season. This reversal is consistent with theory suggesting that colonies decrease production in males if fitness of late emerging males is low. To further investigate reproductive decisions within colonies, we determined if the male mates of multiply-mated queens varied in their reproductive success over time. Sperm use by queens did vary over time suggesting that male success may depend on sperm clumping within the female reproductive tract. Finally, we tested if colony sex ratio conformed to expectations under kin selection theory that nestmate relatedness would positively correlate with investment in new queens if workers controlled sex allocation. Surprisingly, the proportion of queens produced by colonies was negatively correlated with nestmate relatedness, suggesting that allocation may be shaped by advantages arising from increased genetic diversity resulting from multiple mating by queens. Overall, our study suggests that the reproductive decisions of colonies are flexible and may depend both on environmental cues arising from energetic needs of the colony and genetic cues arising from mating behaviours of queens.

Genetic structure and breeding system in a social wasp and its social parasite.

BACKGROUND: Social insects dominate ecological communities because of their sophisticated group behaviors. However, the intricate behaviors of social insects may be exploited by social parasites, which manipulate insect societies for their own benefit. Interactions between social parasites and their hosts lead to unusual coevolutionary dynamics that ultimately affect the breeding systems and population structures of both species. This study represents one of the first attempts to understand the population and colony genetic structure of a parasite and its host in a social wasp system. RESULTS: We used DNA microsatellite markers to investigate gene flow, genetic variation, and mating behavior of the facultative social parasite Vespula squamosa and its primary host, V. maculifrons. Our analyses of genetic variability uncovered that both species possessed similar amounts of genetic variation and failed to show genetic structure over the sampling area. Our analysis of mating system of V. maculifrons and V. squamosa revealed high levels of polyandry and no evidence for inbreeding in the two species. Moreover, we found no significant differences between estimates of worker relatedness in this study and a previous investigation conducted over two decades ago, suggesting that the selective pressures operating on queen mate number have remained constant. Finally, the distribution of queen mate number in both species deviated from simple expectations suggesting that mate number may be under stabilizing selection. CONCLUSION: The general biology of V. squamosa has not changed substantially from that of a typical, nonparasitic Vespula wasp. For example, population sizes of the host and its parasite appear to be similar, in contrast to other social parasites, which often display lower population sizes than their hosts. In addition, parasitism has not caused the mating behavior of V. squamosa queens to deviate from the high levels of multiple mating that typify Vespula wasps. This stands in contrast to some socially parasitic ants, which revert to mating with few males. Overall, the general similarity of the genetic structure of V. maculifrons and V. squamosa presumably reflects the fact that V. squamosa is still capable of independent colony founding and thus reflects an intermediate stage in the evolution of social parasitism.

The significance of multiple mating in the social wasp Vespula maculifrons.

The evolution of the complex societies displayed by social insects depended partly on high relatedness among interacting group members. Therefore, behaviors that depress group relatedness, such as multiple mating by reproductive females (polyandry), are unexpected in social insects. Nevertheless, the queens of several social insect species mate multiply, suggesting that polyandry provides some benefits that counteract the costs. However, few studies have obtained evidence for links between rates of polyandry and fitness in naturally occurring social insect populations. We investigated if polyandry was beneficial in the social wasp Vespula maculifrons. We used genetic markers to estimate queen mate number in V. maculifrons colonies and assessed colony fitness by counting the number of cells that colonies produced. Our results indicated that queen mate number was directly, strongly, and significantly correlated with the number of queen cells produced by colonies. Because V. maculifrons queens are necessarily reared in queen cells, our results demonstrate that high levels of polyandry are associated with colonies capable of producing many new queens. These data are consistent with the explanation that polyandry is adaptive in V. maculifrons because it provides a fitness advantage to queens. Our research may provide a rare example of an association between polyandry and fitness in a natural social insect population and help explain why queens in this taxon mate multiply.

Irregular brood patterns and worker reproduction in social wasps.

The potential for reproductive conflict among colony members exists in all social insect societies. For example, queens and workers may be in conflict over the production of males within colonies. Kin selection theory predicts that in a colony headed by a multiply-mated queen, worker reproduction is prevented by worker policing in the form of differential oophagy. However, few studies have demonstrated that workers actually lay eggs within queenright colonies. The purpose of this study was to determine if workers laid male eggs within unmanipulated queen-right colonies of the polyandrous social wasps Vespula maculifrons and V. squamosa. We focused our analysis on an unusual brood pattern within colonies, multiple egg cells. We were primarily interested in determining if individuals reared in these irregular circumstances were queen or worker offspring. To address this question, we genotyped 318 eggs from eight V. maculifrons and two V. squamosa colonies. No worker reproduction was detected in any of the queenright colonies; all of the eggs found in multiple egg cells were consistent with being queen-produced. However, the frequency of multiple egg cells differed among colonies, suggesting that queens vary in the frequency of errors they make when laying eggs within cells. Finally, we suggest that workers may not be laying eggs within queenright colonies and that worker reproduction may be controlled through mechanisms other than differential oophagy in polyandrous Vespula wasps.

Lack of conflict during queen production in the social wasp Vespula maculifrons.

Social insects display extreme cooperative and helping behaviours. However, social insect colonies are also arenas of intense competitive interactions. One particularly important matter over which colony members may compete centres on the development of sexual offspring. Specifically, colony members may engage in selfish behaviours leading to reproductive competition, whereby individuals either strive to develop as sexuals or assist kin so that close relatives emerge as new reproductives. We investigated whether reproductive competition occurred in the polyandrous social wasp Vespula maculifrons. We genotyped V. maculifrons workers and new queens at eight polymorphic microsatellite loci to determine if larvae of particular genotypes were reared as gynes more frequently than expected by chance. However, we found no significant evidence of reproductive competition in this species. The proportional contributions of males to workers and new queens did not vary within colonies. Moreover, male reproductive skew did not differ between workers and new queens. Finally, novel statistical techniques uncovered no evidence of patriline reversal, the phenomenon whereby males that contribute little to worker production contribute substantially to new queen production. Consequently, we conclude that individual level selection operating to increase the frequency of selfish behaviours that would lead to reproductive competition has been nullified by colony-level selection acting to maintain colony efficiency and cooperation.