Geographical variation in reproductive character displacement in mate choice by male sailfin mollies

Female Amazon mollies, Poecilia formosa, are a unisexual species that reproduce by gynogenesis. They must coexist and mate with males of other species (usually the mollies Poecilia latipinna or Poecilia mexicana) to induce embryogenesis, but inheritance is strictly maternal. We examined the mating preference of the male sailfin molly, P. latipinna, for female sailfin mollies versus Amazon mollies, P. formosa. We compared the mating preferences of sympatric and allopatric populations collected throughout the Gulf Coast of North America. Male P. latipinna from six populations sympatric with Amazon mollies showed a significantly greater strength of preference for conspecific sailfin females than males from five populations that were allopatric with Amazon mollies. These results provide strong evidence for reproductive character displacement of male mate choice in sympatry. Furthermore, the large geographical range of populations that we tested revealed variation among populations within sympatry and allopatry, indicating that it is important to evaluate a large number of populations when examining reproductive character displacement.     

A Sexual Preference in the Amazon Molly, Poecilia formosa

The Amazon molly, Poecilia formosa, is an all-female species that reproduces by gynogenesis, i.e., it relies on sperm of males of closely related species to trigger embryogenesis. Sperm is supplied by males of P. latipinna and P. mexicana. Amazon mollies live in sympatry with at least one of these species, a few populations live in sympatry with two sperm-donor species. As P. formosa is sperm dependent, it needs mechanisms for species and mate recognition. To investigate the effect of rearing conditions on sexual preferences of Amazon mollies, we raised Amazon mollies in sympatry with P. latipinna and P. mexicana males. We used simultaneous choice tests to determine the effect of age on female sexual preferences. Immature Amazon mollies do not exhibit a preference if given a choice between a P. latipinna and a P. mexicana male, whereas adult P. formosa do have a preference for the P. latipinna male. We used two different stimuli in this study, live males and videotapes of males.     

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.     

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.     

Lack of species discrimination based on chemical cues by male sailfin mollies, <Emphasis Type="Italic">Poecilia latipinna</Emphasis>

When making mating decisions, individuals may rely on multiple cues from either the same or multiple sensory modalities. Although the use of visual cues in sexual selection is well studied, fewer studies have examined the role of chemical cues in mate choice. In addition, few studies have examined how visual and/or chemical cues affect male mating decisions. Male mate choice is important in systems where males must avoid mating with heterospecific females, as is found in a mating complex of Poecilia. Male sailfin mollies, Poecilia latipinna, are sexually parasitized by gynogenetic Amazon mollies, P. formosa. Little is known about the mechanism by which male sailfin mollies base their mating decisions. Here we tested the hypothesis that male sailfin mollies from an allopatric and a sympatric population with Amazon mollies use multiple cues to distinguish between conspecific and heterospecific females. We found that male sailfin mollies recognized the chemical cues of conspecific females, but we found no support for the hypothesis that chemical cues are by themselves sufficient for species discrimination. Lack of discrimination based on chemical cues alone may be due to the close evolutionary history between P. latipinna and P. formosa. Males from populations sympatric with Amazon mollies did not differentially associate with females of either of the two species when given access to both visual and chemical cues of the females, yet males from the allopatric population did associate more with conspecific females than with heterospecific females in the presence of both chemical and visual cues. The lack of discrimination by males from the sympatric population between conspecific and heterospecific females based on both chemical and visual cues suggests that these males require more complex combinations of cues to distinguish species, possibly due to the close relatedness of these species.     

Male Sailfin mollies (Poecilia latipinna) copy the mate choice of other males

Female mate-copying has been shown to occur between heterospecifics:female sailfin mollies Poecilia latipinna copy the choice oftheir gynogenetic associates, Amazon mollies P. formosa. Femalemate-copying thus contributes to the maintenance of this asexual-sexualspecies complex by providing an advantage to male sailfin molliesthat mate with Amazon females; because of mate-copying thesemales increase their attractiveness to conspecific females.Here we show that male mate-copying, an unreported phenomenon,also occurs and that it can reverse male preferences for conspecificfemales. Male mate-copying should also contribute to the maintenanceof gynogens and might be advantageous in allowing males a meansto rapidly assess female receptivity although sometimes resultingin heterospedfic matings.     

PEERING THROUGH THE LOOKING GLASS AT A SEXUAL PARASITE: ARE AMAZON MOLLIES RED QUEENS?

Abstract.— The gynogenetic Amazon molly ( Poecilia formosa ) is a clonal, all-female lineage of livebearing fish that faces an unusual obstacle to evolutionary persistence. Sperm from heterospecific males (either sailfin, P. latipinna , or Atlantic, P. mexicana , mollies) is necessary to trigger embryogenesis. However, none of the male's genes are incorporated into the genome of the gynogenetic offspring. Some investigators have proposed that the evolution of male mate discrimination is a result of this cost, leading to a coevolutionary arms race between male avoidance of P. formosa and P. formosa attractiveness. Given that P. formosa successfully reproduces and has not yet gone extinct, it is clear there are mechanisms by which they attract the sexual attention of males. Although a Red Queen coevolutionary process in typical host/parasite systems has been shown to favor the persistence of sexual species, in this system an arms race has been invoked to explain the reverse. Here I present behavioral data supporting a more parsimonious scenario: that mechanisms of attraction in P. formosa are simply a consequence of its hybrid origin. Poecilia latipinna and P. mexicana males do not discriminate between gynogenetic P. formosa females and first generation sexual hybrid females, and females do not differ in agonistic behaviors associated with competition for mates. Both results contradict predictions from the Red Queen hypothesis. Therefore, coevolution is not necessary to explain the apparent evolutionary persistence of P. formosa.     

Male fish deceive competitors about mating preferences

A fundamental question in animal communication is whether the information provided is honest or deceptive [1, 2]. This problem has received much attention, both in theoretical [1, 3] and experimental [4] work. Here we show that male Atlantic mollies (Poecilia mexicana), when observed during mate choice by another male, reduce their mating activity and no longer prefer mating with one of two females presented, which can be interpreted as an attempt to avoid unintended interception of information by the rival male. Most importantly, focal males directed their first sexual interaction (after they were presented with the rival male) toward the initially nonpreferred female, suggesting that males deceive other males about their mating preferences. Deception by the choosing male may be an adaptation to avoid sperm competition, because surrounding males may use public information and copy the focal male's mate choice.     

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.     

Unusual triploid males in a microchromosome-carrying clone of the Amazon molly, Poecilia formosa

The Amazon molly, Poecilia formosa, is an all-female fish of hybrid origin which reproduces by gynogenesis, i.e. it depends on sperm of males of closely related species to trigger parthenogenetic development of the embryo. Therefore the offspring is clonal and identical to the mother. In rare cases the exclusion mechanism fails and paternal introgression occurs. This may result either in triploid offspring - if the whole haploid chromosome set of the sperm fuses with the diploid egg nucleus - or in siblings with microchromosomes - if only subgenomic amounts of paternal DNA are included. In one of our diploid, microchromosome-carrying laboratory stocks we observed eight triploid individuals which all developed into males. We investigated the mitotic and meiotic chromosomes, the synaptonemal complex (SC), and sperm production of these males, and compared them to males of the gonochoristic parental species (P. latipinna and P. mexicana) and their hybrids. This comparison revealed that P. formosa males are functional males with reduced effective fertility. They show a deviation from the typical 23 bivalents in the synaptonemal complexes as well as in diakinesis due to the triploid state. They produced offspring but only with gynogenetic Amazon molly females. This shows that the probably aneuploid sperm from P. formosa males can trigger parthenogenetic development of unreduced eggs.     

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.     

Misleading mollies: surface- but not cave-dwelling Poecilia mexicana males deceive competitors about mating preferences

Individuals providing misleading information to conspecifics may benefit from deception at the receiver's expense. A recent study (Plath et al., Curr Biol 18:1138–1141, 2008c) suggested that male Atlantic mollies (Poecilia mexicana) deceive rival males about their preferred mate. Here, we contrasted potentially deceptive behavior in surface-dwelling P. mexicana males to males of the cave form of that species (the cave molly). Unlike many other cavefishes, cave mollies have retained functional eyes and readily respond to visual stimuli. Males could interact freely with two females (large and small), and an audience male was visually presented during the second part of the tests. When observed during mate choice, males reduced their mating activity, but this reduction was significantly weaker in cave mollies. Overall, the expression of mating preferences (determined through frequencies of nipping and thrusting) declined in front of an audience; again, this effect was significantly weaker in the cave form. Reduced sexual activity and reduced expression of mating preferences can be interpreted as an attempt of the focal male to avoid unintended interception of information by the rival male. Surface but not cave molly males directed their first sexual interaction (when being observed by the rival male) towards the initially non-preferred female, suggesting that surface-dwelling males deceive rival males about their mating preferences. Deception by the focal males may be an adaptation to avoid sperm competition, since other males in their social environment may use public information and copy the focal male's mate choice. It seems that sending deceptive signals is evolutionarily regressed in the cave molly, since mate choice copying is unlikely to occur under naturally dark conditions, and also the potential to deceive rivals about mating preferences is probably very limited.     

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.     

Turbidity affects association behaviour in male Poecilia latipinna

Male association preferences in a bisexual‐unisexual species complex were studied in clear and turbid environments. In south and central Texas, where the gynogenetic sexual‐parasite Poecilia formosa lives syntopically with Poecilia latipinna as its sexual host species, association times of P. latipinna males with conspecific sexual and heterospecific asexual females in clear and turbid water were measured sequentially. Turbidity had an influence on male mate association behaviour. Males spent less time with any kind of female stimulus in turbid water. There was no preference for conspecific sexual females, either in turbid water or under clear conditions. Also, origin of males and acclimatization to turbid water had no effect. How turbidity as a source of visual noise might affect communication among individuals and how this environmental factor might contribute to the stability of this sexual‐asexual mating complex in nature are discussed.     

Mating success of male guppy <Emphasis Type="Italic">Poecilia reticulata</Emphasis> in the wild

To understand how the diversity among male sexual traits is maintained despite directional female choice for attractive traits, it is important to study male reproductive success in each reproductive episode. Male reproductive success is indeed the comprehensive result of several processes such as mate choice, mating, and fertilization. In this study, male mating success was estimated in a wild guppy Poecilia reticulata population of Hiji River, Okinawa Island, in southern Japan. The mating success of males was estimated as the percentage of sperm transferred in relation to the total sperm reserve. In a preliminary experiment, it was confirmed that the sperm reserves of males were partly replenished after 12 h and fully replenished after 72 h, when all sperm were released. In addition, the results of investigation in the wild population revealed that approximately 20% of males had no sperm reserves at all, although their sperm reserves should have been replenished by sexual isolation. Male sexual traits such as body size or color pattern did not affect the percentage of sperm transferred. Results from this study provide important insights into male sperm transfer ability and sperm production in this population. However, the high frequency of male sperm-transfer success estimated in this study should to be taken with caution, and whether it is due to differences between wild and laboratory conditions, sex ratio or the rates of multiple matings by females in the wild population remain to be fully explored.     

Size-assortative mating, male choice and female choice in the curculionid beetle Diaprepes abbreviatus

In the beetle Diaprepes abbreviatus (L.) females are larger on average than males, as indicated by elytra length. Size-assortative matings were observed in wild populations in Florida and in laboratory mating experiments. We tested three mechanisms for this size-assortative mating: (1) mate availability; (2) mating constraints; and (3) mate choice. We found that mate choice influenced size-assortative mating by: (1) large and small males preferring to mate with large females; (2) large males successfully competing for large females, leaving small males to mate with small females; and (3) females accepting large males as mates more readily than small males. Males increased their reproductive success by mating with larger, more fecund females. They transferred protein to females during mating. Copyright 1999 The Association for the Study of Animal Behaviour.     

Spectral sensitivity of mollies: comparing surface- and cave-dwelling Atlantic mollies, Poecilia mexicana

The visual pigments of cones and rods in three species of mollies, Poecilia mexicana , Poecilia latipinna and their asexual hybrid Poecilia formosa , were examined using microspectrophotometry. In P. mexicana , populations from extreme photic habitats were used: one population originated from a clear water habitat, one from a milky water habitat and another from a completely dark cave. Ultraviolet-sensitive cones were found in all species. Differences in the λ_max values of the visual pigments were small between species and among the three P. mexicana populations, but dark-reared cave fishes showed appreciably higher variance. The hybrid species P. formosa showed a highly variable long wavelength cone absorbance, ranging from 528·9 to 598·5 nm, suggesting multiple opsin expression or chromophore mixing.     

Male fish use prior knowledge about rivals to adjust their mate choice

Mate choice as one element of sexual selection can be sensitive to public information from neighbouring individuals. Here, we demonstrate that males of the livebearing fish Poecilia mexicana gather complex social information when given a chance to familiarize themselves with rivals prior to mate choice. Focal males ceased to show mating preferences when being observed by a rival (which prevents rivals from copying mating decisions), but this effect was only seen when focal males have perceived rivals as sexually active. In addition, focal males that were observed by a familiar, sexually active rival showed a stronger behavioural response when rivals were larger and thus, more attractive to females. Our study illustrates an unparalleled adjustment in the expression of mating preferences based on social cues, and suggests that male fish are able to remember and strategically exploit information about rivals when performing mate choice.     

Baffling: a condition-dependent alternative mate attraction strategy using self-made tools in tree crickets

Intense sexual selection in the form of mate choice can facilitate the evolution of different alternative reproductive strategies, which can be condition-dependent. Tree cricket males produce long-distance acoustic signals which are used by conspecific females for mate localization and mate choice. Our study shows that baffling, an acoustic call amplification strategy employed by male tree crickets using self-made tools, is a classic example of a condition-dependent alternative strategy. We show that though most males can baffle, less preferred males, such as smaller and lower-amplitude callers, predominantly use this alternative strategy. Baffling allows these males to increase their call amplitude and advertisement range, which attracts a higher number of females. Baffling also gives these males a mating benefit because females mate for longer durations with them. Our results suggest that the advantage of baffling in terms of gain in the number of sperm cells transferred while mating is primarily limited to less preferred males, thus maintaining the polymorphism of calling strategies in the population. We summarize that baffling is a condition-dependent strategy used by less preferred tree cricket males to obtain mating benefits.     

Multiple post‐mating barriers to hybridization in field crickets

Mechanisms that prevent different species from interbreeding are fundamental to the maintenance of biodiversity. Barriers to interspecific matings, such as failure to recognize a potential mate, are often relatively easy to identify. Those occurring after mating, such as differences in the how successful sperm are in competition for fertilisations, are cryptic and have the potential to create selection on females to mate multiply as a defence against maladaptive hybridization. Cryptic advantages to conspecific sperm may be very widespread and have been identified based on the observations of higher paternity of conspecifics in several species. However, a relationship between the fate of sperm from two species within the female and paternity has never been demonstrated. We use competitive microsatellite PCR to show that in two hybridising cricket species, Gryllus bimaculatus and G. campestris, sequential cryptic reproductive barriers are present. In competition with heterospecifics, more sperm from conspecific males is stored by females. Additionally, sperm from conspecific males has a higher fertilisation probability. This reveals that conspecific sperm precedence can occur through processes fundamentally under the control of females, providing avenues for females to evolve multiple mating as a defence against hybridization, with the counterintuitive outcome that promiscuity reinforces isolation and may promote speciation.