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     

Character displacement in sailfin mollies, <Emphasis Type="Italic">Poecilia latipinna</Emphasis>: allozymes and behavior

Synopsis We analyzed variation in allozymes and mating preferences in 12 populations across much of the range of the sailfin molly, Poecilia latipinna. Sailfin mollies can be sympatric with its sexual parasite Amazon mollies, P. formosa. Amazon mollies must co-exist and mate with bisexual males of closely related species (including sailfin mollies) to induce embryogenesis but inheritance is strictly maternal. Where sailfin and Amazon mollies are sympatric there is evidence of reproductive character displacement as males show a significantly stronger mating preference for sailfin molly females over Amazon mollies compared to preferences of males from allopatric populations. From the allozyme data we found a moderate amount of genetic variation across all populations but this variation did not reveal significant partitioning between sympatric and allopatric populations. Additionally, we found no evidence for isolation by distance as genetic distance was not significantly correlated with geographic distance. While allozyme variation also did not significantly correlate with male mating preferences, there was a significant correlation between male mating preferences and geographic distance. This correlation between mating preferences and geographic distance may have arisen from coevolution with Amazon mollies resulting in reproductive character displacement. Taken together, the distribution of genetic and behavioral variation among sympatric and allopatric populations suggests that behavioral evolution has outpaced evolution at the allozyme loci we examined in P. latipinna.     

Are you more than the sum of your parents’ genes? Phenotypic plasticity in a clonal vertebrate and F1 hybrids of its parental species

All known vertebrate clones have originated from hybridization events and some have produced distinct evolutionary lineages via hybrid speciation. Amazon mollies (Poecilia formosa) present an excellent study system to investigate how clonal species have adapted to heterogeneous environments because they are the product of a single hybridization event between male sailfin mollies (Poecilia latipinna) and female Atlantic mollies (Poecilia mexicana). Here, we ask whether the hybrid species differs from the combination of its parental species’ genes in its plastic response to different environments. Using a three-way factorial design, we exposed neonates produced by Amazon mollies and reciprocal F1 hybrid crosses to different thermal (24°C and 29°C) and salinity (0/2, 12, and 20 ppt) regimes. We measured various ontogenetic and life history characteristics across the life span of females. Our major results were as follows: (1) Reaction norms of growth and maturation to temperature and salinity are quite similar between the two hybrid crosses; (2) Amazon molly reaction norms were qualitatively different than the P. latipinna male and P. mexicana female (L×M) hybrids for the ontogenetic variables; (3) Amazon molly reaction norms in reproductive traits were also quite different from L×M hybrids; and (4) The reaction norms of net fertility were very different between Amazon mollies and L×M hybrids. We conclude that best locale for Amazon mollies is not the best locale for hybrids, which suggests that Amazon mollies are not just an unmodified mix of parental genes but instead have adapted to the variable environments in which they are found. Hybridization resulting in asexuality may represent an underappreciated mechanism of speciation because the unlikely events required to produce such hybrids rarely occur and is dependent upon the genetic distance between parental species.     

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.     

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.     

EVOLUTIONARY ORIGIN OF A PARTHENOFORM,THE AMAZON MOLLY POECILIA FORMOSA,ON THE BASIS OF A MOLECULAR GENEALOGY

The appearance of vertebrate species that reproduce without genetic recombination has been explained by their origin from a rare hybridization event between members of two distantly related species. For the first recognized vertebrate unisexual, the Amazon molly Poecilia formosa, mostly morphological and biochemical genetic information has been available so far with respect to its evolutionary origin. DNA sequence analyses of transcribed portions of the genome (tyrosine kinase proto-oncogenes) demonstrated its hybrid state unequivocally. Both alleles can be traced in a DNA sequence-based phylogenetic tree to extant species that represent the parental species or that are closely related to the corresponding extinct forms, namely P. mexicana limantouri and a so far taxonomically ill-defined north Mexican subspecies of the P. latipinna/P. velifera complex. A rough estimate from the mutation rates dates the hybridization event further back than would have been predicted on the basis of “Muller's ratchet” for an ecologically successful species.     

Polymorphic MHC loci in an asexual fish,the amazon molly (Poecilia formosa; Poeciliidae)

Genes of the major histocompatibility complex (MHC) encode molecules that control immune recognition and are highly polymorphic in most vertebrates. The remarkable polymorphisms at MHC loci may be maintained by selection from parasites, sexual selection, or both. If asexual species show equal (or higher) levels of polymorphisms at MHC loci as sexual ones, this would mean that sexual selection is not necessary to explain the high levels of diversity at MHC loci. In this study, we surveyed the MHC diversity of the asexual amazon molly (Poecilia formosa) and one of its sexual ancestors, the sailfin molly (P. latipinna), which lives in the same habitat. We found that the asexual molly has polymorphic MHC loci despite its clonal reproduction, yet not as polymorphic as the sexual species. Although the nucleotide diversity was similar between the asexual and sexual species, the sexual species exhibited a greater genotypic diversity compared to the asexual one from the same habitats. Within‐genome diversity was similar for MHC class I loci, but for class IIB, the sexual species had higher diversity compared to the asexual — despite the hybrid origins and higher levels of heterozygosity at microsatellite loci in the asexual species. The level of positive selection appears to be similar between the two species, which suggests that these polymorphisms are maintained by selection. Thus, our findings do not allow us to rule out the sexual selection hypothesis for the evolution of MHC diversity, and although the sexual fish has higher levels of MHC‐diversity compared to the asexual species, this may be due to differences in demography, parasites, or other factors, rather than sexual selection.     

Differences in thermal tolerance in coexisting sexual and asexual mollies (Poecilia, Poeciliidae, Teleostei)

This study reports significant differences between the gynogenetic Amazon molly Poecilia formosa and one of its sperm hosts, and the sexual sailfin molly Poecilia latipinna in the critical temperatures at which individual fishes lost motion control. Based on these measurements, it is suggested that cold snaps occurring in winter, but not summer temperatures, can significantly change population composition of these closely related fishes by inflicting higher mortality on P. formosa .     

Frozen F1's amidst a masterpiece of nature: new insights into the rare hybrid origin of gynogenesis in the Amazon molly (Poecilia formosa)

All‐female ‘species’ of fish have been shown to be great models in ecological and evolutionary studies because of the insights they can provide into the origin and evolution of asexuality, the ecology of hybrids, associations between genotype and environment, and the maintenance of sex. Gynogenetic organisms that evolved from sexual ancestors, and combine the disadvantageous traits from sexuality and asexuality, have long baffled evolutionary biologists trying to understand their origin and persistence with their sympatric sexual counterparts. In this issue, a new study using an integrated molecular phylogenetic and classical genetic approach has uncovered compelling evidence regarding the obscure asexual origin of the Amazon molly, Poecilia formosa. By performing an extensive phylogeographic analysis, Stöck et al. (2010) provide evidence that the Amazon molly arose only once within its history, with monophyly being strongly supported by mitochondrial DNA and microsatellite analyses. This result, combined with an elaborate failed attempt to resynthesize the lineage, suggests that vertebrate gynogens such as the Amazon molly are not rare because they are at a disadvantage to their sexual counterparts, but because the genomic conditions under which they arise are rare. Organisms that apparently combine the disadvantages of both sexuality and asexuality remain difficult to understand from both an ecological and an evolutionary perspective, and Stöck et al. (2010) highlight several outstanding important questions. Nonetheless, given that we now have a better knowledge of the origin and history of this unique ‘species’, this should allow researchers to better understand how these frozen F1’s can persist amidst the masterpiece of nature.     

Aspects of the mating behaviour of male mollies (Poecilia spp.)

The unisexual fish Poecilia formosa (the Amazon molly) reproduces by gynogenesis, a process in which sperm from the males of the host bisexual species activates development of its eggs. Unisexuals live with one of the host species in nature and compete with bisexual females for the males. It was long thought that male discrimination and mate selection established a balance between the unisexual and bisexual populations. Thus, hierarchies of males were set up in which dominant males mated with their conspecific females and subordinates mated with the Amazon molly. Recent evidence suggested, however, that male fish do not discriminate between their own females and the Amazon molly, and that there always are more males available than sexually receptive females of both species.
Our findings indicate that male behaviour may be more complex than suggested by either hypothesis. Mate discrimination and courtship behaviour appear to increase with age, so that large males show almost complete preference for their conspecific females, but smaller males will mate with the Amazon molly. In complex groups, small males often dart in and mate with their own females whilst the large males are engaged in courtship activities and defence of territory.     

Dietary niche overlap in sympatric asexual and sexual livebearing fishes Poecilia spp

The present study investigated the spatiotemporal patterns in trophic resource use in a system of a gynogenetic poeciliid fish, the Amazon molly Poecilia formosa, and its sexual congeners the sailfin molly Poecilia latipinna and the Atlantic molly Poecilia mexicana using gut contents analysis. No statistically significant differences in trophic resource use were found between sexual and gynogenetic species, but gut contents varied significantly across sites and over time. In addition, variation in trophic morphology (i.e. gut length) was significant across sites but not species, and laboratory experiments indicated that gut length is phenotypically plastic. Overall, trophic differentiation between coexisting asexual and sexual Poecilia appears to be minimal, and it is unlikely that niche differentiation contributes to a stable coexistence of the two reproductive forms.     

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.     

The role of female competition in the mating success of two clones of the Amazon molly, Poecilia formosa

Clones of the unisexual fish Poecilia formosa , the Amazon molly, live in the wild in association with their progenitor bisexual species, and compete with them for resources. These resources include food, space and also mates, since the Amazon molly reproduces gynogenetically, a process in which sperm transferred from a male host species activates their eggs. Earlier studies of the adaptive mechanisms that facilitate the coexistence of the bisexual and unisexual forms have focused upon male behaviour, but recent work with another unisexual species, Poeciliopsis , demonstrated that female aggressive behaviour may play an important part in determining the ecological standing of a clone.
We have extended our observations to female mating behaviour in two clones in the Amazon molly; clone 1 that does not thrive well under laboratory culture; and clone 2 that is hardy and survives very well. There was no aggression between the clones, nor did the clones show aggression towards females of the bisexual species. However, clone 1 consistently was more successful in the number of mating encounters made with the male; it approached the male more often than clone 2 and fled from the male less often. Males also showed some preference for mating with clone 1 females. If there are physiological differences in the wild clones similar to those seen in the laboratory, the mating advantage of the less robust clone 1 may ensure its continued survival in competition with a more vigorous clone.     

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.     

Limia vittata as host species for the Amazon molly: no evidence for sexual reproduction

DNA fingerprinting demonstrated no sexual reproduction of the gynogenetic Amazon molly Poecilia formosa with Limia vittata as host species, in contrast with a single report that claims to have found sexual reproduction with such matings.     

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.     

The impact of social context on male mate preference in a unisexual--bisexual mating complex

Male sailfin mollies Poecilia latipinna were tested in five different treatments that varied in the relative frequency of heterospecific gynogens (Amazon molly Poecilia formosa) to conspecific females to determine whether social interactions among males within a population causes some males to mate with heterospecific females. Male P. latipinna inseminated a significantly higher proportion of conspecific females and fertilized a significantly higher number of conspecific eggs regardless of the treatment. Nonetheless, preference for conspecific females was not exclusive as a range of 20 to 50% of heterospecific females were fertilized. Social interactions among males may best explain the results and may therefore play an important role in the maintenance of unisexual--bisexual mating complexes.     

Isolation and characterization of 43 microsatellite DNA markers for guppy (Poecilia reticulata)

Thirty‐nine polymorphic microsatellite DNA markers were isolated from the guppy (Poecilia reticulata) genomic library. All of the loci showed moderate allelic variation ranging from two to seven alleles, with observed heterozygosities from 0.000 to 0.938. The microsatellite DNA markers isolated will be available for use in analysis of quantitative trait loci in breeding programmes and for population genetic studies on experimental fish.     

Development of 51 genomic microsatellite DNA markers of guppy (Poecilia reticulata) and their application in closely related species

Fifty‐one microsatellite DNA markers of guppy (Poecilia reticulata) were developed. All of the markers detected moderate allelic variation. The number of alleles for each locus ranged from two to 10. Observed and expected heterozygosities varied from 0.10 to 0.63 and from 0.23 to 0.77, respectively. Three additional fish species of family Poeciliidae were used for the cross‐species amplification of these markers. It was found that only four to 16 markers detected polymorphism in these three fish species.     

A somatic mosaic of the gynogenetic Amazon molly

Breeding experiments with the gynogenetic Amazon molly, Poecilia formosa , proved that mosaicism, detected by flow cytometry with the fluorescent dye DAPI was restricted to the soma, and that gynogenetic reproduction was maintained by the offspring. Potential pathways for how mosaicism might have arisen in P. formosa are discussed.