The effect of rearing temperature on body shape and meristic characters in zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) juveniles

Considering its immediate costs of producing dispensable males, the maintenance of sexual reproduction is a major paradox in evolutionary biology. Asexual lineages that do not face such costs theoretically should replace sexuals over time. Nonetheless, several systems are known in which closely related sexual and asexual lineages stably coexist. In the present study, we studied a sexual/asexual mating complex of a sperm-dependent parthenogenetic fish (amazon molly, Poecilia formosa) and its sexual congeners, the sailfin molly P. latipinna and the Atlantic molly P. mexicana. We asked whether differences in feeding behavior could contribute to their stable coexistence. We conducted a laboratory experiment to compare feeding efficiencies and also measured the competitive abilities between the two reproductive forms. Additionally, we measured gut fullness of fishes caught in natural habitats. Contrary to our predictions, we could not find P. formosa to be less efficient in feeding. We argue that food competition in mollies plays a minor role in mediating coexistence between closely related asexual and sexual mollies.     

How to go extinct by mating too much: population consequences of male mate choice and efficiency in a sexual–asexual species complex

Selection acting on individuals is not predicted to maximize population persistence, yet examples that explicitly quantify conflicts between individual and population level benefits are scarce. One such conflict occurs over sexual reproduction because of the cost of sex: sexual populations that suffer the cost of producing males have only half the growth rate compared to asexuals. Male behaviour can additionally impact population dynamics in a variety of ways, and here we study an example where the impact is unusually clear: the riddle of persistence of sperm‐dependent sexual–asexual species complexes. Here, a sexually reproducing host species coexists with an ameiotically reproducing all‐female sperm parasite. Sexual–asexual coexistence should not be stable because the proportion of asexually reproducing females will rapidly increase and the relative abundance of the sexually reproducing host species will decline. A severe shortage of males will lead to sperm limitation for sexual and asexual females and the system collapses. Male mate choice could reduce the reproductive potential of the asexual species and thus potentially prevent the collapse. In the gynogenetic (sperm‐dependent parthenogenetic) Amazon molly Poecilia formosa and its host (P. latipinna or P. mexicana), males discriminate against asexual females to some extent. Using a population‐dynamical model, we examine the population dynamics of this species complex with varying strengths of male discrimination ability and efficiency with which they locate females and produce sperm. The sexual species would benefit from stronger discrimination, thus preventing being displaced by the asexual females. However, males would be required to evolve preferences that are probably too strong to be purely based upon selection acting on individuals. We conclude that male behaviour does not fully prevent but delays extinction, yet this is highly relevant because low local extinction rates strongly promote coexistence as a metapopulation.     

Divergent female mating preference in a clonal fish

In asexual all-female species, such as the Amazon molly, Poecilia formosa, that depend on sperm from “host males” only to trigger embryogenesis, mate choice does not provide any indirect, genetic benefits to the choosing female, although direct benefits are possible. Asexual species are thought to have a low evolutionary potential or evolvability due to the absence of meiotic recombination. Hence, theory predicts that mating preferences in P. formosa for male ornaments—if existent—should resemble those of females of the two parental species (Poecilia latipinna and Poecilia mexicana) involved in the natural hybridization that gave rise to the asexual P. formosa. When examining the female preference for dummy males with or without black vertical bars in the two parental species and in two lineages of P. formosa, a preference was detected in P. latipinna, but not in P. mexicana females. Interestingly, P. formosa living syntopic with P. latipinna also preferred striped males, while others living syntopic with P. mexicana preferred non-striped males. The evolutionary significance of this phenomenon remains largely unexplained, but it might indicate the evolution of mating preferences in a species with low evolutionary potential. Possible mechanisms include introgression and mitotic gene conversion. Females might use male coloration as indicator mechanisms for male traits that matter in terms of direct benefits.     

Examination of boldness traits in sexual and asexual mollies (<Emphasis Type="Italic">Poecilia latipinna</Emphasis>, <Emphasis Type="Italic">P. formosa</Emphasis>)

Considering the high costs of sexual reproduction (e.g., the production of males), its maintenance and predominance throughout the Animal Kingdom remain elusive. Especially the mechanisms allowing for a stable coexistence of closely related sexual and asexual species are still subject to a lively debate. Asexuals should rapidly outnumber sexuals due to higher population growth rates, unless they face some disadvantages. Here, we investigate potential differences in feeding behavior in a system of sexual (sailfin mollies, Poecilia latipinna) and coexisting gynogenetic fishes (Amazon mollies, Poecilia formosa). In two different experiments, we tested for differences in behavioral traits associated with boldness. Bold individuals take higher risks for gains in resources, so shyer individuals should be less competitive. Our study was motivated by the recent finding that P. formosa are less likely to be preyed upon by piscine predators than P. latipinna. We asked whether this result is indicative of low boldness in P. formosa. However, no differences between the two species were detectible in our behavioral experiments measuring (a) time to emerge from shelter to explore a novel environment, (b) latency time until feeding in a novel environment, and (c) recovery time until feeding restarted after a simulated predator attack. Furthermore, different boldness measures were not correlated with each other within individuals.     

Mating Preferences of the Gynogenetic Amazon Molly Differ Between Populations Sympatric with Different Host Species

Gynogenetic species rely on sperm from heterospecifics for reproduction but do not receive genetic benefits from mating because none of the paternal genome is incorporated into offspring. The gynogenetic Amazon molly (Poecilia formosa) is a species of hybrid origins that are sympatric with one of the two parent species that provide sperm for reproduction, P. latipinna or P. mexicana. Amazons should not prefer to mate with one species over the other because sperm from both species will trigger embryogenesis, but mating preferences may be present in Amazons through other mechanisms. Amazons may prefer the more familiar species (males found in sympatry), or Amazons may prefer males with the greatest lateral projection area (LPA), a preference that is present in the parent species and may be retained within the Amazon hybrid genome. We tested association preferences of two populations of Amazons sympatric with either P. mexicana or P. latipinna. We first performed live trials to test whether Amazons preferred one host species over the other and found that neither population of Amazons showed a preference. We then tested whether Amazons preferred sympatric male (familiar) host or the male with the greatest lateral projection area (LPA) using four animated male models that varied in host species and manipulation of LPA. We found Amazons from a population sympatric with P. latipinna showed no variation in their association preference across the different models. In contrast, Amazons from a population sympatric with P. mexicana (naturally small LPA) spent more time associating with the male models that had smaller LPA, which is more familiar to this population of Amazons. We suggest that Amazons may have population differences in mating preferences, where Amazons sympatric with P. latipinna may not show mating preference for host species, but Amazons sympatric with P. mexicana may show preferences for more familiar‐shaped males.     

Juvenile survival in a unisexual/sexual complex of mollies

Unisexual species like the gynogenetic Amazon molly, Poecilia formosa, enjoy a twofold advantage over sexual species, because they do not produce males. Therefore, unisexuals should be able to outcompete and consequently, replace sexual species. For sperm-dependent (gynogenetic) unisexuals this creates a paradox: they cannot replace their sexual hosts without eradicating themselves. Thus, mechanisms must be in place to stabilize such mating systems. We assessed juvenile survivorship between asexual P. formosa and sexual Poecilia latipinna as a possible factor allowing for persistence and coexistence between the two sympatric species. Offspring of gynogenetic Amazon mollies did not differ significantly in survivorship compared to their sexual host, the Sailfin molly, P. latipinna. The presence of an adult female significantly reduced survival in both species, suggesting that filial cannibalisms operates in this system, but does not appear to play a role in stabilizing mixed sexual/asexual populations. Clark Hubbs, who spent 59 years at the University of Texas and was widely regarded as one of the state’s foremost researchers in the field of ichthyology, the study of fish, passed away February 3rd of 2008 after a long battle with colorectal cancer. He was 86.     

ORIGIN AND MAINTENANCE OF A FEMALE MATING PREFERENCE

We investigated the role of maintenance and origin of female mating preferences in three species of fish. Poecilia latipinna and P. mexicana are sexual species that hybridized to form the gynogenetic clonal P. formosa, which reproduces asexually but requires sperm to initiate embryogenesis. We demonstrate that all three species display almost identical and statistically indistinguishable preferences for large males. Although processes of good genes, runaway sexual selection, and direct selection could maintain preferences in the sexual species, good genes and runaway sexual selection are unlikely to operate in the asexual species. Furthermore, we found that the most likely direct selection benefit, an increase in fecundity, can also be excluded in the gynogens. We conclude that the most parsimonious explanation for this P. formosa preference is that it was inherited from the parental species and is maintained without forces generated by good genes, runaway selection, or direct selection for increased fecundity. This preference may be maintained because of pleiotropic effects (e.g., sensory bias) or mate searching costs.     

Differential susceptibility to food stress in neonates of sexual and asexual mollies (<Emphasis Type="Italic">Poecilia</Emphasis>, Poeciliidae)

The maintenance of sex is still an evolutionary puzzle given its immediate costs. Stably coexisting complexes of asexually and sexually reproducing forms allow to study mechanisms that balance the costs and benefits of both asexual and sexual reproduction. Here, we tested whether coexisting asexual and sexual fish of the genus Poecilia differed in neonate mortality when exposed to environmental stress in the form of fluctuating temperatures and food deprivation. We find that asexual Amazon mollies, Poecilia formosa, are significantly more sensitive to food stress than their sexual relative Poecilia latipinna, but both are equally unaffected by variable temperatures. Differences in the susceptibility to environmental stress may contribute to diminishing the asexuals’ benefits of a higher intrinsic population growth rate and thus mediate stable coexistence of the two reproductive forms.     

Discordance between nuclear and mitochondrial genomes in sexual and asexual lineages of the freshwater snail Potamopyrgus antipodarum

The presence and extent of mitonuclear discordance in coexisting sexual and asexual lineages provides insight into 1) how and when asexual lineages emerged, and 2) the spatial and temporal scales at which the ecological and evolutionary processes influencing the evolution of sexual and asexual reproduction occur. Here, we used nuclear single‐nucleotide polymorphism (SNP) markers and a mitochondrial gene to characterize phylogeographic structure and the extent of mitonuclear discordance in Potamopyrgus antipodarum. This New Zealand freshwater snail is often used to study the evolution and maintenance of sex because obligately sexual and obligately asexual individuals often coexist. While our data indicate that sexual and asexual P. antipodarum sampled from the same lake population are often genetically similar, suggesting recent origin of these asexuals from sympatric sexual P. antipodarum, we also found significantly more population structure in sexuals vs. asexuals. This latter result suggests that some asexual lineages originated in other lakes and/or in the relatively distant past. When comparing mitochondrial and nuclear population genetic structure, we discovered that one mitochondrial haplotype (‘1A’) was rare in sexuals, but common and widespread in asexuals. Haplotype 1A frequency and nuclear genetic diversity were not associated, suggesting that the commonness of this haplotype cannot be attributed entirely to genetic drift and pointing instead to a role for selection.     

Existence of two sexual races in the planarian species switching between asexual and sexual reproduction

In certain planarian species that are able to switch between asexual and sexual reproduction, determining whether a sexual has the ability to switch to the asexual state is problematic, which renders the definition of sexuals controversial. We experimentally show the existence of two sexual races, acquired and innate, in the planarian Dugesia ryukyuensis. Acquired sexuals used in this study were experimentally switched from asexuals. Inbreeding of acquired sexuals produced both innate sexuals and asexuals, but inbreeding of innate sexuals produced innate sexuals only and no asexuals. Acquired sexuals, but not innate sexuals, were forced to become asexuals by ablation and regeneration (asexual induction). This suggests that acquired sexuals somehow retain asexual potential, while innate sexuals do not. We also found that acquired sexuals have the potential to develop hyperplastic and supernumerary ovaries, while innate sexuals do not. In this regard, acquired sexuals were more prolific than innate sexuals. The differences between acquired and innate sexuals will provide a structure for examining the mechanism underlying asexual and sexual reproduction in planarians.     

Biogeography of the Amazon molly: ecological niche and range limits of an asexual hybrid species

Aim To understand the relative contributions of environmental factors, dispersal limitations and the presence of sperm donors in determining the distribution of the Amazon molly (Poecilia formosa), a sperm‐dependent unisexual fish species of hybrid origin. To explore niche similarities and/or differences between the hybrid and parental species. To evaluate whether large‐scale abiotic factors can explain a successful introduction of both P. formosa and Poecilia latipinna. Location South‐east United States, Mexico and Central America. Methods We used abiotic variables in ecological niche modelling (ENM) to identify regions with suitable conditions for the presence of the Amazon molly and its two parental species (P. latipinna and Poecilia mexicana). We also used a recently developed metric to calculate the degree of niche overlap between the hybrid and its parental species. Results ENM produced highly significant models [all area under the curve (AUC) > 0.99 for the three species]. Annual mean temperature and minimum temperature of the coldest month were the variables that best explained the distribution of the Amazon molly. With the exception of south Florida, few areas beyond the known distribution of the species were predicted to have suitable environmental conditions. The hybrid species niche overlaps partially with the parental species. However, given the available data, it is neither more similar nor more different than expected by chance. Main conclusions Two different processes are acting to limit the distribution of P. formosa. At the northern limit, although a sperm donor species is present further north, suitable environmental conditions are absent from nearby locations. At the southern limit, a sperm donor species is present and areas with good environmental conditions are present at nearby locations, suggesting that dispersal ability is the limiting factor. We found that the hybrid species overlaps in a similar way with both parental species while still having its own niche identity. This result may be explained by the fact that hybrid species inherit characteristics of two ecologically divergent species, which can result in intermediate or even transgressive phenotypes. These results support recent work on the role of hybridization in diversification.     

Novel distribution pattern between coexisting sexual and obligate asexual variants of the true estuarine macroalga Ulva prolifera

Where sexual and asexual forms coexist within a species, the asexuals are often found to be prevalent in marginal habitats. This asexual distribution pattern has received evolutionary attention linked to the paradox of sex. In many marine species, there is a distributional trend of asexual modes being more common in lower salinity waters regarded as the ecogeographic marginal, being explained by negative effects of low salinities on sexual reproductive success. However, the distribution pattern of estuarine species recently adapted to low salinity waters has remained unknown. The brackish macroalga Ulva prolifera being a major benthic component of estuarine ecosystems includes a sexual variant and obligate asexual variants by means of motile spores. We examined the sexual–asexual distribution pattern of this alga along a salinity gradient in river estuaries. Surprisingly, opposite to the distributional trend of marine organisms, the results clearly showed the persistent predominance of sexuals in the lower salinity reaches than the asexuals. In addition, a frequent alternating of dioecious gametophytes and sporophytes in the sexual population was observed, suggesting the sexual reproductive process would be robustly performed in the low salinity waters. Considering U. prolifera had evolved from the ancestral marine species to become a true estuarine species of which the core habitat is the low salinity reaches, in a broad sense its sexual–asexual distribution pattern would be involved in asexual marginal occupations of the species range previously reported in other organisms. Based on the frozen niche variation model, we can give a concise explanation for the evolutionary process of this pattern; multiple asexuals with frozen genotypic variation had arisen from sexual ancestors undergoing low salinity adaptation and share the estuarine habitat with the sexuals at present.     

Effects of Female‐Female Aggression in a Sexual/Unisexual Species Complex

The maintenance of sexual reproduction is still a largely unresolved question in evolutionary biology, and one of the most puzzling aspects of this is the coexistence of sexual and asexual females. This often leads to widespread niche overlap, posing an interesting challenge to competitive exclusion theory. In this study, we investigate how the aggressive behaviors between females in a sexual/unisexual mating complex of mollies (Poecilia latipinna and P. formosa) differ, and the effects these behaviors have on both the performer and receiver of aggressive acts. We exposed females to five treatments: (1) alone (control); (2) a large and small sexual female; (3) a large sexual female and a small unisexual female; (4) a small sexual female and a large unisexual female; and (5) a large and small unisexual female. We found that P. formosa females were on average more aggressive over time when compared to their sexual hosts, P. latipinna. The females of P. formosa, however, incurred a higher cost of being aggressive. Large, aggressor females of this species had a lower body fat content than their conspecific recipients. By contrast, P. latipinna females showed no differentiation between aggressors and recipients. Here, we combine life history methodology and behavioral observations to show that female competition can in fact be quantitatively measured and that these aggressive behaviors are costly to perform. The adaptive value of this interspecies aggression is not yet completely understood, but sexual competition, access to resources, and position within the shoal are all possible functions.     

HYBRIDIZATION LEADS TO SENSORY REPERTOIRE EXPANSION IN A GYNOGENETIC FISH,THE AMAZON MOLLY (POECILIA FORMOSA): A TEST OF THE HYBRID‐SENSORY EXPANSION HYPOTHESIS

Expansions in sensory systems usually require processes such as gene duplication and divergence, and thus evolve slowly. We evaluate a novel mechanism leading to rapid sensory repertoire expansion: hybrid‐sensory expansion (HSE). HSE occurs when two species with differently tuned sensory systems form a hybrid, bringing together alleles from each of the parental species. In one generation, a sensory repertoire is created that is the sum of the variance between parental species. The Amazon molly presents a unique opportunity to test the HSE hypothesis in a “frozen” hybrid. We compared opsin sequences of the Amazon molly, Poecilia formosa, to those of the parental species. Both parental species are homozygous at the RH2–1 locus and each of the four long wavelength sensitive loci, while P. formosa possess two different alleles at these loci; one matching each parental allele. Gene expression analysis showed P. formosa use the expanded opsin repertoire that was the result of HSE. Additionally, behavioral tests revealed P. formosa respond to colored stimuli in a manner similar or intermediate to the parental species P. mexicana and P. latipinna. Together these results strongly support the HSE hypothesis. Hybrid‐sensory repertoire expansion is likely important in other hybrid species and in other sensory systems.     

Do I Know You? Species Recognition Operates Within and Between the Sexes in a Unisexual–Bisexual Species Complex of Mollies

Male sailfin mollies (Poecilia latipinna) can be sexually parasitized by closely related, unisexual, gynogenetic Amazon mollies (Poecilia formosa). This study examined possible cues used by male P. latipinna to distinguish between conspecific females and sympatric, heterospecific P. formosa. Digital photos were used to create models to test male P. latipinna preference for model female P. latipinna and P. formosa with a full suite of traits and altered models of P. latipinna and P. formosa. Male P. latipinna significantly preferred models of either species over no stimulus, demonstrating that models elicit a male response. Males also significantly preferred female P. latipinna models over P. formosa models. We also examined species recognition by female sailfin mollies using the same models, and found that female sailfin mollies significantly preferred to associate with female P. latipinna over P. formosa. These results taken together suggest that the use of fish models yield results similar to those studies using live stimuli. Male preference was then tested for unaltered vs. altered models in the following combinations: (i) P. formosa vs. P. formosa with a female P. latipinna fin; (ii) P. formosa vs. P. formosa with a female P. latipinna lateral spot pattern; (iii) P. formosa vs. P. latipinna with a P. formosa fin and their spotless lateral pattern. Males did not significantly prefer models with any isolated traits over the unaltered P. formosa models. Thus, males may be using traits other than the ones isolated for species recognition or males may be using a suite of multiple traits to recognize conspecific females.     

Female sperm limitation in natural populations of a sexual/asexual mating complex (Poecilia latipinna, Poecilia formosa)

In sperm-dependent sexual/asexual mating systems, male mate choice is critical for understanding the mechanisms behind apparent stability observed in natural populations. The gynogenetic Amazon molly (Poecilia formosa) requires sperm from sexual males (e.g. Poecilia latipinna) to trigger embryogenesis, but inheritance is strictly maternal. Consequently, males should try to avoid or reduce the cost of mating with asexuals. We investigated male mate choice by documenting the presence of sperm in natural populations and found that a higher proportion of sexual females had sperm than asexuals. In addition, among those females that had sperm, sexuals had more sperm than asexuals. Our results hint at a role for male mate choice as a stabilizing factor in such systems.     

Behavioural and life‐history regulation in a unisexual/bisexual mating system: does male mate choice affect female reproductive life histories?

In theory, unisexual taxa have an advantage over ecologically similar bisexual species because unisexuals produce twice as many daughters and, thus, should quickly outcompete coexisting bisexuals in any given population. For sperm‐dependent unisexual (gynogenetic) species, stable coexistence with their bisexual sperm donors can be postulated if male mate choice puts unisexual females at a disadvantage through sperm limitation, thus halving their reproductive output compared to bisexuals (‘behavioural regulation hypothesis’). We tested for a potential life‐history signature of male mate choice in a system of coexisting bisexual sailfin mollies (Poecilia latipinna) and gynogenetic Amazon mollies (Poecilia formosa). Specifically, we gave P. latipinna males an opportunity to freely interact (and mate) with both types of females and, after 25 days, quantified the proportion of (1) females with sperm in their genital tract and (2) pregnant females. A higher proportion of P. latipinna females (53.7%) had sperm in their genital tract (compared to only 25.9% in P. formosa), corroborating a previous study on wild‐caught fish. This translated into a higher frequency (42.6%) of P. latipinna females being pregnant (compared to 29.6% in P. formosa); however, among pregnant females, no significant differences between species in reproductive life‐history traits (such as offspring number or size) were uncovered. Hence, although the findings of the present study confirm that male discrimination against unisexual females leads to reduced reproductive output in unisexuals, the observed magnitude of differences in targeted life histories between the two types of females is unlikely to be the sole factor regulating stable coexistence in this system. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 598–606.     

Feeding rates in the sailfin molly Poecilia latipinna and its coexisting sexual parasite,the gynogenetic Amazon molly Poecilia formosa

Feeding rates of the gynogenetic Amazon molly Poecilia formosa and one of its sexual hosts, the sailfin molly Poecilia latipinna, were measured under winter and summer temperature conditions. Food consumption of the unisexual P. formosa in winter conditions was significantly higher than that of P. latipinna, and it is hypothesized that the resulting food stress might have an important influence on the population composition of these closely related fishes via higher winter mortality in P. formosa.     

Male mating behaviour of a molly, <Emphasis Type="Italic">Poecilia latipunctata</Emphasis>: a third host for the sperm-dependent Amazon molly, <Emphasis Type="Italic">Poecilia formosa</Emphasis>

The Tamesí molly, Poecilia latipunctata, has a very limited biogeographical range in northeast Mexico. This area is nested within the ranges of the Atlantic molly, Poecilia mexicana, and the unisexual Amazon molly, Poecilia formosa. Based on morphology, especially fin shape, the Tamesí molly has been considered to be a "short-fin" molly. We describe the courtship sequence of P. latipunctata. The courtship clearly places the species into the clade of "long-fin" mollies, a finding that corroborates earlier studies based on nuclear DNA and mitochondrial DNA. All three species live together in certain habitats. This renders P. latipunctata a potential host species for the sperm-dependent, unisexual Amazon molly. Using behavioural tests, we demonstrate that P. latipunctata males actually copulate with Amazon mollies, despite a pronounced preference for conspecific females. In laboratory experiments P. latipunctata males are capable of triggering embryogenesis in P. formosa females. Field observations support the hypothesis that P. latipunctata is a third host species for P. formosa, indicating that the Amazon molly effectively exploits all available host species for its gynogenetic mode of reproduction. Electronic Publication     

VARIATION IN THE STRENGTH OF MALE MATE CHOICE ALLOWS LONG‐TERM COEXISTENCE OF SPERM‐DEPENDENT ASEXUALS AND THEIR SEXUAL HOSTS

In several asexual taxa, reproduction requires mating with related sexual species to stimulate egg development, even though genetic material is not incorporated from the sexuals (gynogenesis). In cases in which gynogens do not invest in male function, they can potentially have a twofold competitive advantage over sexuals because the asexuals avoid the cost of producing males. If unmitigated, however, the competitive success of the asexuals would ultimately lead to their own demise, following the extinction of the sexual species that stimulate egg development. We have studied a model of mate choice among sexual individuals and asexual gynogens, where males of the sexual species preferentially mate with sexual females over gynogenetic females, to determine if such mating preferences can stably maintain both gynogenetic and sexual individuals within a community. Our model shows that stable coexistence of gynogens and their sexual hosts can occur when there is variation among males in the degree of preference for mating with sexual females and when pickier males pay a higher cost of preference.