Spatial and temporal escape from fungal parasitism in natural communities of anciently asexual bdelloid rotifers

Sexual reproduction is costly, but it is nearly ubiquitous among plants and animals, whereas obligately asexual taxa are rare and almost always short-lived. The Red Queen hypothesis proposes that sex overcomes its costs by enabling organisms to keep pace with coevolving parasites and pathogens. If so, the few cases of stable long-term asexuality ought to be found in groups whose coevolutionary interactions with parasites are unusually weak. In theory, antagonistic coevolution will be attenuated if hosts disperse among patches within a metapopulation separately from parasites and more rapidly. We examined whether these conditions are met in natural communities of bdelloid rotifers, one of the longest-lived asexual lineages. At any life stage, these microscopic invertebrates can tolerate the complete desiccation of their ephemeral freshwater habitats, surviving as dormant propagules that are readily carried by the wind. In our field experiments, desiccation and wind transport enabled bdelloids to disperse independently of multiple fungal parasites, in both time and space. Surveys of bdelloid communities in unmanipulated moss patches confirmed that fungal parasitism was negatively correlated with extended drought and increasing height (exposure to wind). Bdelloid ecology therefore matches a key condition of models in which asexuals persist through spatio-temporal decoupling from coevolving enemies.     

Antagonistic Coevolution and Sex

One of the leading hypotheses for the maintenance of sexual reproduction is the Red Queen hypothesis. The underlying premise of the Red Queen hypothesis is that parasites rapidly evolve to infect common host genotypes. This response by parasites could result in the long-term maintenance of genetic variation and may favor sexual reproduction over asexual reproduction. The underlying ideas present a wonderful microcosm for teaching evolution. Here I present the reasons for why sex is anomalous for evolutionary theory, the rationale underlying the Red Queen hypothesis, and some empirical studies of the Red Queen hypothesis using a freshwater snail. The empirical results are consistent with the Red Queen hypothesis. In addition, the distribution of sexual and asexual reproduction in the snail leads naturally to thinking about coevolution in a geographic mosaic of parasite-mediated natural selection.     

Bloody‐minded parasites and sex: the effects of fluctuating virulence

Asexual lineages can grow at a faster rate than sexual lineages. Why then is sexual reproduction so widespread? Much empirical evidence supports the Red Queen hypothesis. Under this hypothesis, coevolving parasites favour sexual reproduction by adapting to infect common asexual clones and driving them down in frequency. One limitation, however, seems to challenge the generality of the Red Queen: in theoretical models, parasites must be very virulent to maintain sex. Moreover, experiments show virulence to be unstable, readily shifting in response to environmental conditions. Does variation in virulence further limit the ability of coevolving parasites to maintain sex? To address this question, we simulated temporal variation in virulence and evaluated the outcome of competition between sexual and asexual females. We found that variation in virulence did not limit the ability of coevolving parasites to maintain sex. In fact, relatively high variation in virulence promoted parasite‐mediated maintenance of sex. With sufficient variation, sexual females persisted even when mean virulence fell well below the threshold virulence required to maintain sex under constant conditions. We conclude that natural variation in virulence does not limit the relevance of the Red Queen hypothesis for natural populations; on the contrary, it could expand the range of conditions over which coevolving parasites can maintain sex.     

Escape from the Red Queen: an overlooked scenario in coevolutionary studies

Almost all eukaryotic organisms undergo sexual recombination at some stage of their life history. However, strictly asexual organisms should have higher per capita rate of reproduction compared with those that have sex, so the latter must convey some advantage which overrides the reproductive benefit of asexuality. For example, sexual reproduction and recombination may play an important role in allowing organisms to evolutionarily ‘keep up’ with parasites. Host–parasite coevolution can operate via negative frequency‐dependent selection whereby parasite genotypes adapt to infect host genotypes as they become locally common. By producing more genetically diverse offspring with unique genotypes, sexual organisms have an advantage over asexual counterparts. Essentially, sexual hosts are more difficult for coevolving parasites to ‘track’ over time. This scenario has been named the “Red Queen hypothesis”. It refers to a passage in Lewis Carroll's ‘Through the Looking Glass’ in which the Red Queen tells Alice: ‘it takes all the running you can do, to keep in the same place’; this statement resembles the negative frequency‐dependent dynamics of host–parasite coevolution.     

Parasites in sexual and asexual mollies (Poecilia, Poeciliidae, Teleostei): a case for the Red Queen?

The maintenance of sexual reproduction in the face of its supposed costs is a major paradox in evolutionary biology. The Red Queen hypothesis, which states that sex is an adaptation to fast-evolving parasites, is currently one of the most recognized explanations for the ubiquity of sex and predicts that asexual lineages should suffer from a higher parasite load if they coexist with closely related sexuals. We tested this prediction using four populations of the sexual fish species Poecilia latipinna and its asexual relative Poecilia formosa. Contrary to expectation, no differences in parasite load could be detected between the two species.     

The maintenance of sex: host–parasite coevolution with density‐dependent virulence

Why don’t asexual females replace sexual females in most natural populations of eukaryotes? One promising explanation is that parasites could counter the reproductive advantages of asexual reproduction by exerting frequency‐dependent selection against common clones (the Red Queen hypothesis). One apparent limitation of the Red Queen theory, however, is that parasites would seem to be required by theory to be highly virulent. In the present study, I present a population‐dynamic view of competition between sexual females and asexual females that interact with co‐evolving parasites. The results show that asexual populations have higher carrying capacities, and more unstable population dynamics, than sexual populations. The results also suggest that the spread of a clone into a sexual population could increase the effective parasite virulence as population density increases. This combination of parasite‐mediated frequency‐dependent selection, and density‐dependent virulence, could lead to the coexistence of sexual and asexual reproductive strategies and the long‐term persistence of sex.     

Role of parasite load and differential habitat preferences in maintaining the coexistence of sexual and asexual competitors in fish of the Cobitis taenia hybrid complex

In the context of the paradoxical ubiquity of sex, we tested whether stable coexistence of sexual and asexual fish of the genus Cobitis is mediated by parasites, as asexual fish suffer more from parasitic infections because of their lower genetic variability [the Red Queen hypothesis (RQH)], or by partial niche shift of the two strains differing in mode of reproduction. We did not find a clear correlation between infection risk with a helminth parasite and the proportion of sexuals, and we found similar infection rates among sexual females and co‐occurring asexuals in general, including the most frequent clone in particular. These results suggest that the mechanisms of the RQH are not directly engaged in stabilizing this asexual complex. On the other hand, the temporally stable gradient in sexual/asexual proportions along the river correlated with gradients in environmental parameters (physicochemical water parameters, velocity, and shading of the habitat) and turnover in the fish assemblage structure. Sexual and asexual forms thus appear to prefer different habitats. The Cobitis teania asexual complex thus contributes to the view that persistence of sex may, as in many taxa, be driven by case‐specific processes. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 220–235.     

On the reality and recognisability of asexual organisms: morphological analysis of the masticatory apparatus of bdelloid rotifers

Species concepts and definitions have been a long-standing debate in evolutionary biology since before Darwin, and almost all proposed solutions are based upon grouping and clustering, with species conceived as somehow biological distinct entities, originated and maintained mainly by reproductive isolation. Lacking reproductive exchange, asexual organisms such as bdelloid rotifers, the best-supported clade of so-called 'ancient asexuals', pose an interesting challenge to debates over the reality of species. However, few data are available on bdelloid rotifers. The only evidence has been that bdelloid species have been more consistently recognised than in their sister sexual group, the monogonont rotifers, across successive taxonomic treatments, but this is confounded by the much lesser degree of taxonomic interest in bdelloids. We applied geometric morphometrics analyses on shape and size of hard masticatory pieces, named trophi, of 1420 bdelloids, belonging to 48 populations of eight traditional species, to test the hypothesis of recognisability of bdelloids. Our morphological analysis confirms that traditional bdelloid species are separated distinct entities, possessing trophi morphologies divided by gaps between taxa, similar to patterns of morphological features in sexually reproducing organisms. In common with most microscopic understudied organisms, bdelloid rotifers harbour much previously undescribed diversity: we found significant differences in trophi morphology within traditional species, revealing the existence of cryptic taxa, similar to those also found in facultatively sexual monogonont rotifers. We confirm that recognisability in bdelloids is not qualitatively different from other small understudied animals such as monogononts, and that sexual versus asexual reproduction does not lead to differences in morphological diversity patterns, as previously suggested based on interpretation of taxonomic revisions.     

Sex versus parasitism versus density

The Red Queen hypothesis predicts that sexual reproduction should be favoured when the risk of infection by parasites is high. However, this prediction may also be achieved independently by means of the reproductive assurance and the density-dependent transmission hypotheses because higher densities increase the chances of fertilization (i.e. mates are easier to find) and increase the risk of infection. Additionally, the Red Queen hypothesis makes a long-term prediction that infection should be genotype-specific, with time-lagged dynamics. In the present study, we attempt to consolidate these two predictions by investigating whether sexuality and parasitism alternate in a time-lagged fashion on an ecological time-scale. In a 3-year study, we examined the time-lagged dynamics between male frequencies, infection levels (by Centrocestus sp.), and snail densities of four natural populations of sexual and asexual Melanoides tuberculata snails. The results obtained provide no evidence of either prediction of the Red Queen hypothesis. We found partial support for reproductive assurance in a single population, and could not discriminate between the three hypotheses in another population. The remaining two populations did not support any of the hypotheses in question.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 93 , 537–544.     

HOST-PARASITE COEVOLUTION: EVIDENCE FOR RARE ADVANTAGE AND TIME-LAGGED SELECTION IN A NATURAL POPULATION

In theory, parasites can create time-lagged, frequency-dependent selection in their hosts, resulting in oscillatory gene-frequency dynamics in both the host and the parasite (the Red Queen hypothesis). However, oscillatory dynamics have not been observed in natural populations. In the present study, we evaluated the dynamics of asexual clones of a New Zealand snail, Potamopyrgus antipodarum, and its trematode parasites over a five-year period. During the summer of each year, we determined host-clone frequencies in random samples of the snail to track genetic changes in the snail population. Similarly, we monitored changes in the parasite population, focusing on the dominant parasite, Microphallus sp., by calculating the frequency of clones in samples of infected individuals from the same collections. We then compared these results to the results of a computer model that was designed to examine clone frequency dynamics for various levels of parasite virulence. Consistent with these simulations and with ideas regarding dynamic coevolution, parasites responded to common clones in a time-lagged fashion. Finally, in a laboratory experiment, we found that clones that had been rare during the previous five years were significantly less infectible by Microphallus when compared to the common clones. Taken together, these results confirm that rare host genotypes are more likely to escape infection by parasites; they also show that host-parasite interactions produce, in a natural population, some of the dynamics anticipated by the Red Queen hypothesis.     

Parasite transmission among relatives halts Red Queen dynamics

The theory that coevolving hosts and parasites create a fluctuating selective environment for one another (i.e., produce Red Queen dynamics) has deep roots in evolutionary biology; yet empirical evidence for Red Queen dynamics remains scarce. Fluctuating coevolutionary dynamics underpin the Red Queen hypothesis for the evolution of sex, as well as hypotheses explaining the persistence of genetic variation under sexual selection, local parasite adaptation, the evolution of mutation rate, and the evolution of nonrandom mating. Coevolutionary models that exhibit Red Queen dynamics typically assume that hosts and parasites encounter one another randomly. However, if related individuals aggregate into family groups or are clustered spatially, related hosts will be more likely to encounter parasites transmitted by genetically similar individuals. Using a model that incorporates familial parasite transmission, we show that a slight degree of familial parasite transmission is sufficient to halt coevolutionary fluctuations. Our results predict that evidence for Red Queen dynamics, and its evolutionary consequences, are most likely to be found in biological systems in which hosts and parasites mix mainly at random, and are less likely to be found in systems with familial aggregation. This presents a challenge to the Red Queen hypothesis and other hypotheses that depend on coevolutionary cycling.     

Phylogenomics of Unusual Histone H2A Variants in Bdelloid Rotifers

Rotifers of Class Bdelloidea are remarkable in having evolved for millions of years, apparently without males and meiosis. In addition, they are unusually resistant to desiccation and ionizing radiation and are able to repair hundreds of radiation-induced DNA double-strand breaks per genome with little effect on viability or reproduction. Because specific histone H2A variants are involved in DSB repair and certain meiotic processes in other eukaryotes, we investigated the histone H2A genes and proteins of two bdelloid species. Genomic libraries were built and probed to identify histone H2A genes in Adineta vaga and Philodina roseola, species representing two different bdelloid families. The expressed H2A proteins were visualized on SDS-PAGE gels and identified by tandem mass spectrometry. We find that neither the core histone H2A, present in nearly all other eukaryotes, nor the H2AX variant, a ubiquitous component of the eukaryotic DSB repair machinery, are present in bdelloid rotifers. Instead, they are replaced by unusual histone H2A variants of higher mass. In contrast, a species of rotifer belonging to the facultatively sexual, desiccation- and radiation-intolerant sister class of bdelloid rotifers, the monogononts, contains a canonical core histone H2A and appears to lack the bdelloid H2A variant genes. Applying phylogenetic tools, we demonstrate that the bdelloid-specific H2A variants arose as distinct lineages from canonical H2A separate from those leading to the H2AX and H2AZ variants. The replacement of core H2A and H2AX in bdelloid rotifers by previously uncharacterized H2A variants with extended carboxy-terminal tails is further evidence for evolutionary diversity within this class of histone H2A genes and may represent adaptation to unusual features specific to bdelloid rotifers.     

Geographic variation in sterilizing parasite species and the Red Queen

The Red Queen hypothesis predicts that sexual reproduction should be favoured in locations where the risk of infection by virulent parasites is consistently high. When hosts are exposed to multiple parasites over their geographic range, the coevolving parasite species may vary among host populations. We surveyed 26 streams on the South Island of New Zealand to determine whether the frequency of snails ( Potamopyrgus antipodarum ) infected by various sterilizing trematode parasite species was correlated with the frequency of sexual individuals. We compared the results with a survey conducted over 20 years ago to determine whether the associations were consistent. We also evaluated different measures of parasite-mediated selection among populations, including prevalence of the most common local parasite (MCLP) species and parasite diversity to assess the best predictor of sexual reproduction among stream populations. The results showed that the relationship between male frequency and parasite infection is more geographically widespread than previously recorded. Additionally, we found that the prevalence of the MCLP was the best predictor of sex in habitats where hosts populations are infected with multiple parasites (approximately 15 trematode species). This study provides evidence that sexual snails occur more often in environments with high infection levels, and that the pattern of parasite-imposed selection is geographically variable. Support for the Red Queen may be strengthened by focussing on the MCLP, which may vary among host populations.     

Parasites driving host diversity: Incidence of disease correlated with Daphnia clonal turnover*

According to the Red Queen hypothesis, clonal diversity in asexual populations could be maintained by negative frequency‐dependant selection by coevolving parasites. If common clones are selected against and rare clones gain a concomitant advantage, we expect that clonal turnover should be faster during parasite epidemics than between them. We tested this hypothesis exploring field data of the Daphnia–Caullerya host–parasite system. The clonal make‐up and turnover of the Daphnia host population was tracked with high temporal resolution from 1998 until 2013, using first allozyme and later microsatellite markers. Significant differences in the clonal composition between random and infected subsamples of Daphnia populations were detected on six of seven tested occasions, confirming genetic specificity of the host–parasite interaction in this system. We used time series analysis to compare the rates of host clonal turnover to the incidence of parasitism, and found that Caullerya prevalence was significantly associated with microsatellite‐based clonal turnover. As alternate hypotheses, we further tested whether turnover was related to a variety of biotic, abiotic, and host demographic parameters. Other significant correlates of turnover were cyanobacterial biomass and (weakly) temperature. Overall, parasitism seems to be a strong driver of host clonal turnover, in support of the Red Queen hypothesis.     

Bdelloid rotifers: ‘sleeping beauties’ and ‘evolutionary scandals’, but not only

Bdelloid rotifers are mostly known for two peculiarities, continuous parthenogenetic reproduction and dormancy in response to habitat desiccation, a phenomenon named anhydrobiosis. These uncommon traits earned them the names of ‘evolutionary scandals’ and ‘sleeping beauties’, respectively. Relevant aspects of bdelloid biology have recently been described that connect parthenogenesis to anhydrobiosis and that might account for their evolutionary survival in spite of the conservative reproduction. In the present study, I explore recent literature, in the attempt to disentangle the apparent incongruency between the apomictic reproduction and the presumed long-term evolutionary survival of bdelloid species. Recent results remarkably improved our knowledge of bdelloid population biology, genetics, and molecular biology. The most relevant findings concern (i) acquisition of foreign genes through horizontal transfer, (ii) presence of divergent sequences possibly corresponding to ancient gene duplications and (iii) capacity to escape parasites: events that appear to be connected with dormancy. I also address the results of recent studies on the relationships between bdelloids and other rotifers, including acanthocephalans, in an attempt to highlight similarities and differences that should be clarified to better understand phylogenetic relationship among the Rotifera sensu lato.

     

Immediate and heritable costs of desiccation on the life history of the bdelloid rotifer <Emphasis Type="Italic">Philodina roseola</Emphasis>

The long-term persistence of the ‘ancient asexual’ bdelloid rotifers, a clade of small aquatic invertebrates, is often tied to their ability to enter anhydrobiosis. This ability has both clear benefits (e.g. survival of desiccating conditions), but offers considerable costs (e.g. subsequent repair of the genome as well as physiological and metabolic costs to re-establish the phenotype). Despite these costs, several studies show that the time spent dry is effectively ignored with respect to life expectancy (the Sleeping Beauty hypothesis) and that reconstruction of the genome after a desiccation event might even be necessary to repair mistakes accumulated in it from obligate parthenogenesis while the animals were active. We propose that this genomic repair might not derive exclusively because desiccation per se, but could also result from genetic exchange that appears to occur between individuals during this time. By comparing individuals of Philodina roseola Ehrenberg, 1832 desiccated in groups versus individually, we document costs to desiccation in the isolated treatment group that impact negatively on lifespan and reproduction. In addition, comparing both groups with continuously active individuals reveals no strong evidence for the Sleeping Beauty hypothesis in this species nor any decline in fitness over a six-month period for the latter group. Finally, many treatment effects are at least partly heritable and were found in the untreated F1 generations. In particular, individuals desiccated in groups and their offspring could both reproduce faster than the offspring of continuously active individuals. Thus, our results offer additional support for the hypothesis of genetic exchange occurring during desiccation events in P. roseola and highlight the importance of considering this factor, and desiccation in general, in explaining bdelloid fitness. Moreover, our results provide additional context for understanding how the genetic information of bdelloids is ultimately shaped.     

Diversity and the maintenance of sex by parasites

The Red Queen hypothesis (RQH) predicts that parasite‐mediated selection will maintain sexual individuals in the face of competition from asexual lineages. The prediction is that sexual individuals will be difficult targets for coevolving parasites if they give rise to more genetically diverse offspring than asexual lineages. However, increasing host genetic diversity is known to suppress parasite spread, which could provide a short‐term advantage to clonal lineages and lead to the extinction of sex. We test these ideas using a stochastic individual‐based model. We find that if parasites are readily transmissible, then sex is most likely to be maintained when host diversity is high, in agreement with the RQH. If transmission rates are lower, however, we find that sexual populations are most likely to persist for intermediate levels of diversity. Our findings thus highlight the importance of genetic diversity and its impact on epidemiological dynamics for the maintenance of sex by parasites.     

Faster clonal turnover in high‐infection habitats provides evidence for parasite‐mediated selection

According to the Red Queen hypothesis for sex, parasite‐mediated selection against common clones counterbalances the reproductive advantage of asexual lineages, which would otherwise outcompete sexual conspecifics. Such selection on the clonal population is expected to lead to a faster clonal turnover in habitats where selection by parasites is stronger. We tested this prediction by comparing the genetic structure of clonal and sexual populations of freshwater snail Potamopyrgus antipodarum between years 2003 and 2007 in three depth‐specific habitats in Lake Alexandrina (South Island, New Zealand). These habitats differ in the risk of infection by castrating trematodes and in the relative proportion of sexual individuals. As predicted, we found that the clonal structure changed significantly in shallow and mid‐water habitats, where prevalence of infection was high, but not in the deep habitat, where parasite prevalence was low. Additionally, we found that both clonal diversity and evenness of the asexual population declined in the shallow habitat. In contrast, the genetic structure (based on F–statistics) of the coexisting sexual population did not change, which suggests that the change in the clonal structure cannot be related to genetic changes in the sexual population. Finally, the frequency of sexuals had no effect on the diversity of the sympatric clonal population. Taken together, our results show a more rapid clonal turnover in high‐infection habitats, which gives support for the Red Queen hypothesis for sex.     

SEXUAL SPECIES ARE SEPARATED BY LARGER GENETIC GAPS THAN ASEXUAL SPECIES IN ROTIFERS

Why organisms diversify into discrete species instead of showing a continuum of genotypic and phenotypic forms is an important yet rarely studied question in speciation biology. Does species discreteness come from adaptation to fill discrete niches or from interspecific gaps generated by reproductive isolation? We investigate the importance of reproductive isolation by comparing genetic discreteness, in terms of intra‐ and interspecific variation, between facultatively sexual monogonont rotifers and obligately asexual bdelloid rotifers. We calculated the age (phylogenetic distance) and average pairwise genetic distance (raw distance) within and among evolutionarily significant units of diversity in six bdelloid clades and seven monogonont clades sampled for 4211 individuals in total. We find that monogonont species are more discrete than bdelloid species with respect to divergence between species but exhibit similar levels of intraspecific variation (species cohesiveness). This pattern arises because bdelloids have diversified into discrete genetic clusters at a faster net rate than monogononts. Although sampling biases or differences in ecology that are independent of sexuality might also affect these patterns, the results are consistent with the hypothesis that bdelloids diversified at a faster rate into less discrete species because their diversification does not depend on the evolution of reproductive isolation.