Does high relatedness promote cheater-free multicellularity in synthetic lifecycles?

The evolution of multicellularity is one of the key transitions in evolution and requires extreme levels of cooperation between cells. However, even when cells are genetically identical, noncooperative cheating mutants can arise that cause a breakdown in cooperation. How then, do multicellular organisms maintain cooperation between cells? A number of mechanisms that increase relatedness amongst cooperative cells have been implicated in the maintenance of cooperative multicellularity including single-cell bottlenecks and kin recognition. In this study, we explore how relatively simple biological processes such as growth and dispersal can act to increase relatedness and promote multicellular cooperation. Using experimental populations of pseudo-organisms, we found that manipulating growth and dispersal of clones of a social amoeba to create high levels of relatedness was sufficient to prevent the spread of cheating mutants. By contrast, cheaters were able to spread under low-relatedness conditions. Most surprisingly, we saw the largest increase in cheating mutants under an experimental treatment that should create intermediate levels of relatedness. This is because one of the factors raising relatedness, structured growth, also causes high vulnerability to growth rate cheaters.

Social amoebae mating types do not invest unequally in sexual offspring.

Unequal investment by different sexes in their progeny is common and includes differential investment in the zygote and differential care of the young. The social amoeba Dictyostelium discoideum has a sexual stage in which isogamous cells of any two of the three mating types fuse to form a zygote which then attracts hundreds of other cells to the macrocyst. The latter cells are cannibalized and so make no genetic contribution to reproduction. Previous literature suggests that this sacrifice may be induced in cells of one mating type by cells of another, resulting in a higher than expected production of macrocysts when the inducing type is rare and giving a reproductive advantage to this social cheat. We tested this hypothesis in eight trios of field-collected clones of each of the three D. discoideum mating types by measuring macrocyst production at different pairwise frequencies. We found evidence that supported differential contribution in only two of the 24 clone pairs, so this pattern is rare and clone-specific. In general, we did not reject the hypothesis that the mating types contribute cells relative to their proportion in the population. We also found a significant quadratic relationship between partner frequency and macrocyst production, suggesting that when one clone is rare, macrocyst production is limited by partner availability. We were also unable to replicate previous findings that macrocyst production could be induced in the absence of a compatible mating partner. Overall, mating type-specific differential investment during sex is unlikely in microbial eukaryotes like D. discoideum.

Sex ratio and gamete size across eastern North America in Dictyostelium discoideum, a social amoeba with three sexes.

Theory indicates that numbers of mating types should tend towards infinity or remain at two. The social amoeba, Dictyostelium discoideum, however, has three mating types. It is therefore a mystery how this species has broken the threshold of two mating types, but has not increased towards a much higher number. Frequency-dependent selection on rare types in combination with isogamy, a form of reproduction involving gametes similar in size, could explain the evolution of multiple mating types in this system. Other factors, such as drift, may be preventing the evolution of more than three. We first looked for evidence of isogamy by measuring gamete size associated with each type. We found no evidence of size dissimilarities between gametes. We then looked for evidence of balancing selection, by examining mating type distributions in natural populations and comparing genetic differentiation at the mating type locus to that at more neutral loci. We found that mating type frequency varied among the three populations we examined, with only one of the three showing an even sex ratio, which does not support balancing selection. However, we found more population structure at neutral loci than the mating type locus, suggesting that the three mating types are indeed maintained at intermediate frequencies by balancing selection. Overall, the data are consistent with balancing selection acting on D. discoideum mating types, but with a sufficiently weak rare sex advantage to allow for drift, a potential explanation for why these amoebae have only three mating types.

Concurrent coevolution of intra-organismal cheaters and resisters.

The evolution of multicellularity is a major transition that is not yet fully understood. Specifically, we do not know whether there are any mechanisms by which multicellularity can be maintained without a single-cell bottleneck or other relatedness-enhancing mechanisms. Under low relatedness, cheaters can evolve that benefit from the altruistic behaviour of others without themselves sacrificing. If these are obligate cheaters, incapable of cooperating, their spread can lead to the demise of multicellularity. One possibility, however, is that cooperators can evolve resistance to cheaters. We tested this idea in a facultatively multicellular social amoeba, Dictyostelium discoideum. This amoeba usually exists as a single cell but, when stressed, thousands of cells aggregate to form a multicellular organism in which some of the cells sacrifice for the good of others. We used lineages that had undergone experimental evolution at very low relatedness, during which time obligate cheaters evolved. Unlike earlier experiments, which found resistance to cheaters that were prevented from evolving, we competed cheaters and noncheaters that evolved together, and cheaters with their ancestors. We found that noncheaters can evolve resistance to cheating before cheating sweeps through the population and multicellularity is lost. Our results provide insight into cheater-resister coevolutionary dynamics, in turn providing experimental evidence for the maintenance of at least a simple form of multicellularity by means other than high relatedness.

A conserved extraordinarily long serine homopolymer in Dictyostelid amoebae.

Eukaryotic protein sequences often contain amino-acid homopolymers that consist of a single amino acid repeated from several to dozens of times. Some of these are functional but others may persist largely because of high expansion rates due to DNA slippage. However, very long homopolymers with over a hundred repeats are very rare. We report an extraordinarily long homopolymer consisting of 306 tandem serine repeats from the single-celled eukaryote Dictyostelium discoideum, which also has a multicellular stage. The gene has a paralog with 132 repeats and orthologs, also with high serine repeat numbers, in various other Dictyostelid species. The conserved gene structure and protein sequences suggest that the homopolymer is functional. The high codon diversity and very poor alignment of serine codons in this gene between species similarly indicate functionality. This is because the serine homopolymer is conserved despite much DNA sequence change. A survey of other very long amino-acid homopolymers in eukaryotes shows that high codon diversity is the rule, suggesting that these too may be functional.

On status badges and quality signals in the paper wasp Polistes dominulus: body size, facial colour patterns and hierarchical rank.

To establish a dominance order, social animals often rely on indicators of fighting to avoid costly aggressive encounters. In some species, individuals use colour patterns to signal their social status. Recent studies claimed that facial markings in the eusocial paper wasp Polistes dominulus are status badges that allow co-foundresses to form a linear hierarchy based on individual quality. Here, we evaluated facial patterns in natural populations of P. dominulus, in its native range, to observe whether the marks reflect overall wasp quality in different contexts. We used the same measures of clypeus patterns used by earlier studies, but did not find that they functioned as status badges. Our analyses showed no evidence that visual markers are related to: (i) size, (ii) probability of surviving winter, (iii) social rank in spring associations, or (iv) health status (assessed by the presence of strepsipteran endoparasites). Size, however, is important. Larger wasps are more likely to survive the winter and to acquire the dominant position in spring associations. Larvae infected with endoparasites become smaller adult wasps. These findings suggest that body size is a reliable quality indicator on which wasps build their social networks, and that clypeus patterning is not involved.

The social parasite wasp Polistes atrimandibularis does not form host races.

Parasites that exploit the parental behaviour of several host species may be selected to form distinct host-specific genetic lineages. This process is well documented in bird brood parasites, but not in insect social parasites. Polistes atrimandibularis is the only paper-wasp social parasite known to exploit four host species. It does not form genetically distinct host races according to analyses based on microsatellite loci. Also, there were no size-matching between parasites and host species. Instead, P. atrimandibularis queens seemed to be successful as parasites in this population only when they originated from nests of P. dominulus, the largest species. The other host species are a sink for P. atrimandibularis since adult females emerging from those nests appear too small to usurp colonies themselves. Traits that may help P. atrimandibularis infiltrate multiple species may include its nonaggressive usurpation tactics and its ability to acquire host cuticular hydrocarbon recognition labels.

A linear dominance hierarchy among clones in chimeras of the social amoeba Dictyostelium discoideum.

Amoebae from different clones of Dictyostelium discoideum aggregate into a common slug, which migrates towards light for dispersal, then forms a fruiting body consisting of a somatic, dead stalk, holding up a head of living spores. Contributions of two clones in a chimera to spore and stalk are often unequal, with one clone taking advantage of the other's stalk contribution. To determine whether there was a hierarchy of exploitation among clones, we competed all possible pairs among seven clones and measured their relative representation in the prespore and prestalk stages and in the final spore stage. We found a clear linear hierarchy at the final spore stage, but not at earlier stages. These results suggest that there is either a single principal mechanism or additive effects for differential contribution to the spore, and that it involves more than spore/stalk competition.

Worker interests and male production in Polistes gallicus, a Mediterranean social wasp.

The resolution of social conflict in colonies may accord with the interests of the most numerous party. In social insect colonies with single once-mated queens, workers are more closely related to the workers' sons than they are to the queens' sons. Therefore, they should prefer workers to produce males, against the queen's interests. Workers are capable of producing males as they arise from unfertilized eggs. We found Polistes gallicus to have colonies of single, once-mated queens, as determined by microsatellite genotyping of the workers, so worker interests predict worker male production. In colonies lacking queens, workers produced the males, but not in colonies with original queens. Thus worker interests were expressed only when the queen was gone. The high fraction of missing queens and early end to the colony cycle relative to climate so early in the season is surprising and may indicate a forceful elimination of the queen.

Co-occurrence in nature of different clones of the social amoeba, Dictyostelium discoideum.

The social amoeba, Dictyostelium discoideum, produces a multicellular fruiting body and has become a model system for cell-cell interactions such as signalling, adhesion and development. However, unlike most multicellular organisms, it forms by aggregation of cells and, in the laboratory, forms genetic chimeras where there may be competition among clones. Here we show that chimera formation is also likely in nature, because different clones commonly co-occur on a very small scale. This suggests that D. discoideum will likely have evolved strategies for competing in chimeras, and that the function of some developmental genes will be competitive. Natural chimerism also makes D. discoideum a good model organism for the investigation of issues relating to coexistence and conflict between cells.

Swarm-founding in the polistine wasps: the importance of finding many microsatellite loci in studies of adaptation.

We developed 52 microsatellite loci for the wasp, Polybioides tabidus, for the purpose of studying the evolution and inclusive fitness consequences of swarm-founding. The large number of loci is important for three reasons that may apply to many other systems. Heterozygosity was low in our target species, yet we found enough polymorphic loci for accurate kinship studies in this species. Many monomorphic loci were polymorphic in other polistine wasps, making comparative studies possible. Finally, enough loci amplified over a broad range of species to add a historical dimension. We sequenced six loci in other polistine wasps and used the flanking sequences to construct a phylogeny. Based on this phylogeny, we infer that swarm-founding has evolved independently three times in the polistine wasps.

Absence of within-colony kin discrimination: foundresses of the social wasp, Polistes carolina, do not prefer their own larvae.

There is great potential for conflict within social insect colonies especially when there are multiple inseminated females laying eggs. One reason that conflict is not always realized may be that these females do not identify their own progeny and direct their attentions preferentially towards them. Using DNA microsatellite loci we were able to determine exactly which female was the mother of each larva in eight nests of the social wasp, Polistes carolina. Using 26 h of videotapes of natural nests we observed 2,093 feedings of specific larvae by these adults and found that they did not preferentially feed their own progeny. Instead feedings were distributed to progeny as predicted based on their frequency in the nest. The absence of nepotism towards closest kin within colonies in this system is likely to promote colony harmony.

Unrelated helpers in a social insect.

High-resolution genetic markers have revolutionized our understanding of vertebrate mating systems, but have so far yielded few comparable surprises about kinship in social insects. Here we use microsatellite markers to reveal an unexpected and unique social system in what is probably the best-studied social wasp, Polistes dominulus. Social insect colonies are nearly always composed of close relatives; therefore, non-reproductive helping behaviour can be favoured by kin selection, because the helpers aid reproductives who share their genes. In P. dominulus, however, 35% of foundress nestmates are unrelated and gain no such advantage. The P. dominulus system is unlike all other cases of unrelated social insects, because one individual has nearly complete reproductive dominance over subordinates who could have chosen other reproductive options. The only significant advantage that subordinates obtain is a chance at later reproduction, particularly if the queen dies. Thus, P. dominulus societies are functionally unlike other social insects, but similar to certain vertebrate societies, in which the unrelated helpers gain through inheritance of a territory or a mate.

A phylogenetic perspective on sequence evolution in microsatellite loci.

We examined the evolution of the repeat regions of three noncoding microsatellite loci in 58 species of the Polistinae, a subfamily of wasps that diverged over 140 million years ago. A phylogenetic approach allows two new kinds of approaches to studying microsatellite evolution: character mapping and comparative analysis. The basic repeat structure of the loci was highly conserved, but was often punctuated with imperfections that appear to be phylogenetically informative. Repeat numbers evolved more rapidly than other changes in the repeat region. Changes in number of repeats among species seem consistent with the stepwise mutation model, which is based on slippage during replication as the main source of mutations. Changes in repeat numbers can occur even when there are very few tandem repeats but longer repeats, especially perfect repeats led to greater rates of evolutionary change. Species phylogenetically closer to the one from which we identified the loci had longer stretches of uninterrupted repeats and more different motifs, but not longer total repeat regions. The number of perfect repeats increased more often than it decreased. However, there was no evidence that some species have consistently greater numbers of repeats across loci than other species have, once ascertainment bias is eliminated. We also found no evidence for a population size effect posited by one form of the directionality hypothesis. Overall, phylogenetic variation in repeat regions can be explained by adding neutral evolution to what is already known about the mutation process. The life cycle of microsatellites appears to reflect a balance between growth by slippage and degradation by an essentially irreversible accumulation of imperfections.