DNA EVIDENCE FOR NONHYBRID ORIGINS OF PARTHENOGENESIS IN NATURAL POPULATIONS OF VERTEBRATES

Naturally occurring unisexual reproduction has been documented in less than 0.1% of all vertebrate species. Among vertebrates, true parthenogenesis is known only in squamate reptiles. In all vertebrate cases that have been carefully studied, the clonal or hemiclonal taxa have originated through hybridization between closely related sexual species. In contrast, parthenogenetic reproduction has arisen in invertebrates by a variety of mechanisms, including likely cases of “spontaneous” (nonhybrid) origin, a situation not currently documented in natural populations of vertebrates. Here, we present molecular data from the Neotropical night lizard genus Lepidophyma that provides evidence of independent nonhybrid origins for diploid unisexual populations of two species from Costa Rica and Panama. Our mitochondrial and nuclear phylogenies are congruent with respect to the unisexual taxa. Based on 14 microsatellite loci, heterozygosity (expected from a hybrid origin) is low in Lepidophyma reticulatum and completely absent in unisexual L. flavimaculatum. The unique value of this system will allow direct comparative studies between parthenogenetic and sexual lineages in vertebrates, with an enormous potential for this species to be a model system for understanding the mechanisms of nonhybrid parthenogenesis.     

The origin and evolution of a unisexual hybrid: Poecilia formosa

Clonal reproduction in vertebrates can always be traced back to hybridization events as all known unisexual vertebrates are hybrids between recognized species or genetically defined races. Interestingly, clonal vertebrates often also rely on interspecific matings for their reproduction because gynogenesis (sperm-dependent parthenogenesis) and hybridogenesis are common modes of propagation. While in most cases these hybridization events leave no hereditary traces in the offspring, occasionally the genome exclusion mechanism fails and either small parts of male genetic material remain inside the oocyte in the form of microchromosomes, or fusion of the sperm nucleus with the oocyte nucleus leads to polyploid individuals. In this review, we highlight the important role of hybridization for the origin and evolution of a unisexual hybrid: the Amazon molly, Poecilia formosa.     

Regain of sex determination system and sexual reproduction ability in a synthetic octoploid male fish

Polyploids in vertebrates are generally associated with unisexual reproduction, but the direct consequences of polyploidy on sex determination system and reproduction mode remain unknown. Here, we synthesized a group of artificial octoploids between unisexual gynogenetic hexaploid Carassius gibelio and sexual tetraploid Carassius auratus. The synthetic octoploids were revealed to have more than 200 chromosomes, in which 50 chromosomes including the X/Y sex determination system were identified to transfer from sexual tetraploid C. auratus into the unisexual gynogenetic hexaploid C. gibelio. Significantly, a few synthetic octoploid males were found to be fertile, and one octoploid male was confirmed to regain sexual reproduction ability,which exhibits characteristics that are the same to sexual reproduction tetraploid males, such as 1:1 sex ratio occurrence, meiosis completion and euploid sperm formation in spermatogenesis, as well as normal embryo development and gene expression pattern during embryogenesis. Therefore, the current finding provides a unique case to explore the effect of sex determination system incorporation on reproduction mode transition from unisexual gynogenesis to sexual reproduction along with genome synthesis of recurrent polyploidy in vertebrates.     

Genetic basis and breeding application of clonal diversity and dual reproduction modes in polyploid Carassius auratus gibelio

A unisexual species is generally associated with polyploidy, and reproduced by a unisexual reproduction mode, such as gyno- genesis, hybridogenesis or parthenogenesis. Compared with other unisexual and polyploid species, gibel carp (Carassius au- ratus gibelio) has a higher ploidy level of hexaploid. It has undergone several successive rounds of genome polyploidy, and experienced an additional, more recent genome duplication event. More significantly, the dual reproduction modes, including gynogenesis and s...     

Evolution of sexual mimicry in sperm-dependent all-female forms of Poeciliopsis (Pisces: Poeciliidae)

The twofold advantage of all-female reproduction is limited in many asexual lineages because females are sperm-dependent. Males of a related sexual-host species typically prefer conspecific females as mates. According to the “sexual mimicry” hypothesis, an all-female lineage that closely resembles females of the sexual-host species should have enhanced mating success. Examination of mating success in all-female fish of the genus Poeciliopsis supported this hypothesis. The excellent sexual mimicry of some all-female strains could have evolved through mutations within clonal lineages subsequent to their origins as interspecific hybrids. Alternatively, this mimicry may have been “frozen” from variation in the sexual gene pool when new unisexual lineages first arose. To test the latter hypothesis, we examined laboratory synthesized strains of the hybridogenetic fish P. monacha-lucida. The frozen variation hypothesis was supported by the present results.     

Genetic structure of unisex hybrid population of frogs Rana esculenta complex in lowland Transcarpathian region

The biochemical genetic marking and analysis of the ploidy and sexual structure of the green frog populations from the Transcarpathian lowlands have demonstrated that this region is inhabited by the unisexual populations composed of the allodiploid females only. Their genome includes a small portion of the marsh frog genic diversity. This phenomenon is discussed in relation to the unisexual population reproduction problem. The assumption is proposed that in the Transcarpathian hybrid populations the hybrids are spawned by the parthenogenesis.     

Multiple parthenoforms of Empoasca leafhoppers from Madeira Island: where are these unisexual forms coming from?

There are controversial opinions on whether asexual reproduction is more common on islands than on the mainland. Although some authors consider that the evidences of geographical parthenogenesis support the view that asexual reproduction is more common on islands, comparative data on the modes of reproduction of insular and continental taxa confirming this statement are very limited. In this work, we report the presence of three unisexual forms and three bisexual species of the genus Empoasca (Cicadelloidea, Hemiptera, Insecta) from Madeira Island. Experimentally, the unisexual forms reproduced in the absence of males for several generations. The chromosome analysis has shown that the bisexual species differ from one another in chromosome number, and unisexual forms are apomictic and also each have different chromosome numbers. Of parthenoforms, one is triploid and two are of obscure level of ploidy, 2n or 3n. The results obtained show that for this genus unisexual forms are more common on Madeira Island than in the nearby continental areas. It is suggested that unisexual forms may be more plentiful on islands than on the mainland because if an asexual reproduction event occurs, the relaxing competition in these underexploited and enemy-free habitats may favor the establishment of new parthenogenetic lineages.     

Relating hybrid advantage and genome replacement in unisexual salamanders

Unisexual vertebrates are model systems for understanding the evolution of sex. Many predominantly clonal lineages allow occasional genetic recombination, which may be sufficient to avoid the accumulation of deleterious mutations and parasites. Introgression of paternal DNA into an all-female lineage represents a one-way flow of genetic material. Over many generations, this could result in complete replacement of the unisexual genomes by those of the donor species. The process of genome replacement may be counteracted by contemporary dispersal or by positive selection on hybrid nuclear genomes in ecotones. I present a conceptual model that relates nuclear genome replacement, positive selection on hybrids and biogeography in unisexual systems. I execute an individual-based simulation of the fate of hybrid genotypes in contact with a single host species. I parameterize these models for unisexual salamanders in the Ambystoma genus, for which the frequency of genome replacement has been a source of ongoing debate. I find that, if genome replacement occurs at a rate greater than 1/10,000 in Ambystoma, then there must be compensating positive selection in order to maintain observed levels of hybrid nuclei. Future researchers studying unisexual systems may use this framework as a guide to evaluating the hybrid superiority hypothesis.     

Automictic reproduction in interspecific hybrids of poeciliid fish

Automixis, the process whereby the fusion of meiotic products restores the diploid state of the egg, is a common mode of reproduction in plants but has also been described in invertebrate animals. In vertebrates, however, automixis has so far only been discussed as one of several explanations for isolated cases of facultative parthenogenesis. Analyzing oocyte formation in F1 hybrids derived from Poecilia mexicana limantouri and P. latipinna crosses (the cross that led to the formation of the gynogenetic Poecilia formosa), we found molecular evidence for automictic oocyte production. The mechanism involves the random fusion of meiotic products after the second meiotic division. The fertilization of diploid oocytes gives rise to fully viable triploid offspring. Although the automictic production of diploid oocytes as seen in these F1 hybrids clearly represents a preadaptation to parthenogenetic reproduction, it is also a powerful intrinsic postzygotic isolation mechanism because the resulting next generation triploids were always sterile. The mechanism described here can explain facultative parthenogenesis, as well as varying ploidy levels reported in different animal groups. Most importantly, at least some of the reported cases of triploidy in humans can now be traced back to automixis.     

Preadaptation for parthenogenetic colony foundation in subterranean termites Reticulitermes spp. (Isoptera: Rhinotermitidae)

Thelytokous (all-female producing) parthenogenesis, in some cases, involves reproductive advantages against obligate sexual reproduction. However, the completion of parthenogenesis takes multiple steps without the help of males, and thus preadaptation that meets those requirements will be an important factor for the evolution of parthenogenesis. The Japanese subterranean termite, Reticulitermes speratus, is known to have the ability of parthenogenetic colony foundation, where females that failed to mate with males found colonies cooperatively with partner females and reproduce by parthenogenesis. In this study, we compared the parthenogenetic ability and the colony initiation behavior among six Reticulitermes species in Japan. All species other than R. speratus were not able to reproduce parthenogenetically. Nevertheless, females of these species without the parthenogenetic ability performed homosexual female–female colony initiation and produced eggs without fertilization. In addition, in one species without parthenogenetic reproduction, R. kanmonensis, female–female pair initiated founding behavior as quickly as a heterosexual pair. These results suggest that female–female colony initiation and virgin egg-laying are predominant characters among the genus Reticulitermes and provide a preadaptive condition for parthenogenetic colony foundation in R. speratus.     

A novel nucleo-cytoplasmic hybrid clone formed via androgenesis in polyploid gibel carp

Background

Unisexual vertebrates have been demonstrated to reproduce by gynogenesis, hybridogenesis, parthenogenesis, or kleptogenesis, however, it is uncertain how the reproduction mode contributes to the clonal diversity. Recently, polyploid gibel carp has been revealed to possess coexisting dual modes of unisexual gynogenesis and sexual reproduction and to have numerous various clones. Using sexual reproduction mating between clone D female and clone A male and subsequent 7 generation multiplying of unisexual gynogenesis, we have created a novel clone strain with more than several hundred millions of individuals. Here, we attempt to identify genetic background of the novel clone and to explore the significant implication for clonal diversity contribution.

Methods

Several nuclear genome markers and one cytoplasmic marker, the mitochondrial genome sequence, were used to identify the genetic organization of the randomly sampled individuals from different generations of the novel clone.

Results

Chromosome number, Cot-1 repetitive DNA banded karyotype, microsatellite patterns, AFLP profiles and transferrin alleles uniformly indicated that nuclear genome of the novel clone is identical to that of clone A, and significantly different from that of clone D. However, the cytoplasmic marker, its complete mtDNA genome sequence, is same to that of clone D, and different from that of clone A.

Conclusions

The present data indicate that the novel clone is a nucleo-cytoplasmic hybrid between the known clones A and D, because it originates from the offspring of gonochoristic sexual reproduction mating between clone D female and clone A male, and contains an entire nuclear genome from the paternal clone A and a mtDNA genome (cytoplasm) from the maternal clone D. It is suggested to arise via androgenesis by a mechanism of ploidy doubling of clone A sperm in clone D ooplasm through inhibiting the first mitotic division. Significantly, the selected nucleo-cytoplasmic hybrid female still maintains its gynogenetic ability. Based on the present and previous findings, we discuss the association of rapid genetic changes and high genetic diversity with various ploidy levels and multiple reproduction modes in several unisexual and sexual complexes of vertebrates and even other invertebrates.

     

A TEST OF THE SPONTANEOUS HETEROSIS HYPOTHESIS FOR UNISEXUAL VERTEBRATES

The coupling between clonal modes of reproduction and hybridization in unisexual vertebrates has led to the hypothesis that heterosis accounts for their ecological success (the “spontaneous heterosis” hypothesis). High levels of genic heterozygosity characteristic of unisexual-hybrid vertebrates are believed to result in enhanced growth, survivorship, and fertility relative to their sexual ancestors. To test this hypothesis, we synthesized 33 new unisexual-hybrid strains of fishes in the genus Poeciliopsis (Atheriniformes: Poeciliidae). On average, the synthetic unisexuals had lower survivorship and a higher incidence of birth defects than either of the sexual ancestors or two natural strains of unisexuals. However, a subset of these synthetic unisexuals exhibited characteristics within the range of the sexual ancestors and natural unisexual strains. These results support the alternative hypothesis that the ecological success of natural unisexuals results from selection of the most fit clones from a broad spectrum of genotypes that arose via multiple hybrid events. We propose that the coupling between unisexuality and hybridization in the vertebrates exists because hybridization is a dysgenic process that can disrupt normal gametogenesis and thus lead to clonal reproduction.     

Evolution of the Reproductive Mechanisms in Tardigrades — A Review

Although tardigrades can reproduce only through gametes they have exploited several modes of reproduction, which may be determined by their environment. Marine species (mainly heterotardigrades) are gonochoristic; hermaphroditism is only cited once, and parthenogenesis is unknown. In many cases females mature one egg at a time throughout adult life, whereas males are semelparous. Gonochorism is still present in limno-terrestrial species, while sporadic hermaphroditism occurs in several eutardigrade families. Thelytoky is the most common mode of reproduction in non-marine Tardigrada. Females are iteroparous, laying groups of eggs (free or in the exuvium), while males are semelparous (in a limnic species) or iteroparous with a continuous or cyclical maturation of the spermatozoa (in species from moss and leaf litter). Self-fertilisation appears to characterise hermaphroditic species, found in freshwater, mosses, leaf litter and soil. Egg maturation in these species is similar to that of the gonochoristic species, while spermatozoa mature in appreciable numbers before the oocytes, subsequently maturing continuously but in small numbers over the life of the animal. Parthenogenesis in limno-terrestrial tardigrades always appears continuous. In many species only females occur, but morpho-species populations may be found with both bisexual amphimictic (diploid) and unisexual thelytokous (often but not always polyploid) cytotypes. We can hypothesise that with the evolution of cryptobiosis and passive dispersal unstable and isolated habitats may favour parthenogenesis and self-fertilisation, as both reproductive modes allow colonisation of a new territory by a single individual. Parthenogenesis and hermaphroditism do not occur in the same species, and we can surmise that self-fertilisation will only evolve where parthenogenesis has never occurred.     

Monophyletic origin of multiple clonal lineages in an asexual fish (Poecilia formosa)

Despite the advantage of avoiding the costs of sexual reproduction, asexual vertebrates are very rare and often considered evolutionarily disadvantaged when compared to sexual species. Asexual species, however, may have advantages when colonizing (new) habitats or competing with sexual counterparts. They are also evolutionary older than expected, leaving the question whether asexual vertebrates are not only rare because of their 'inferior' mode of reproduction but also because of other reasons. A paradigmatic model system is the unisexual Amazon molly, Poecilia formosa, that arose by hybridization of the Atlantic molly, Poecilia mexicana, as the maternal ancestor, and the sailfin molly, Poecilia latipinna, as the paternal ancestor. Our extensive crossing experiments failed to resynthesize asexually reproducing (gynogenetic) hybrids confirming results of previous studies. However, by producing diploid eggs, female F(1) -hybrids showed apparent preadaptation to gynogenesis. In a range-wide analysis of mitochondrial sequences, we examined the origin of P. formosa. Our analyses point to very few or even a single origin(s) of its lineage, which is estimated to be approximately 120,000 years old. A monophyletic origin was supported from nuclear microsatellite data. Furthermore, a considerable degree of genetic variation, apparent by high levels of clonal microsatellite diversity, was found. Our molecular phylogenetic evidence and the failure to resynthesize the gynogenetic P. formosa together with the old age of the species indicate that some unisexual vertebrates might be rare not because they suffer the long-term consequences of clonal reproduction but because they are only very rarely formed as a result of complex genetic preconditions necessary to produce viable and fertile clonal genomes and phenotypes ('rare formation hypothesis').     

Reproductive mode of the geographic parthenogenetic mayfly Ephoron shigae, with findings from some new localities (Insecta: Ephemeroptera, Polymitarcyidae)

The burrowing polymitarcyid mayfly Ephoron shigae is distributed widely in Japan. Some populations are bisexual, others are unisexual, and the distributions of the two types overlap broadly. Experimental evidence of parthenogenetic reproduction, long suspected in unisexual populations, is presented here, based on a comparative analysis of the developmental rate of fertilized and unfertilized eggs. The developmental rate of fertilized eggs from 20 mated females in a bisexual population was 98.4% ± 0.73% (mean ± SD), and no unfertilized eggs from 20 virgin females in that population developed. The developmental rates of unfertilized eggs in two unisexual populations were 89.0% ± 4.59% and 84.2% ± 1.96%, respectively. This article presents experimental evidence of geographic parthenogenesis in E. shigae and provides support for the previous interpretation. In addition, we discuss the relationship between the sex ratio of each population and the developmental rate of fertilized versus unfertilized eggs from the females in those populations.     

Parthenogenesis in Hexapoda: Entognatha and non‐holometabolous insects

In hexapods, unlike the majority of animals, development without fertilization is a common phenomenon. They evolved a striking diversity of unisexual reproductive types that include a variety of modes starting from spontaneous parthenogenesis in females to the production of impaternate males with different variants in between. Many reports about parthenogenetic species have accumulated over time. Here, we present a review of various parthenogenetic hexapod groups with a particular focus on their chromosome systems and ploidy level. We show that conclusions about the reproductive mode often lack solid evidence and sometimes inefficiently demonstrate how parthenogenesis is maintained in corresponding groups. In this review, basal hexapods (Protura, Collembola, Diplura), primarily wingless insect groups (‘Apterygota’) and non‐holometabolous insects are listed with references to a variety of their unisexual reproductive modes.     

Unisexual Reproduction Reverses Muller’s Ratchet

Cryptococcus neoformans is a pathogenic basidiomycetous fungus that engages in outcrossing, inbreeding, and selfing forms of unisexual reproduction as well as canonical sexual reproduction between opposite mating types. Long thought to be clonal, >99% of sampled environmental and clinical isolates of C. neoformans are MATα, limiting the frequency of opposite mating-type sexual reproduction. Sexual reproduction allows eukaryotic organisms to exchange genetic information and shuffle their genomes to avoid the irreversible accumulation of deleterious changes that occur in asexual populations, known as Muller’s ratchet. We tested whether unisexual reproduction, which dispenses with the requirement for an opposite mating-type partner, is able to purge the genome of deleterious mutations. We report that the unisexual cycle can restore mutant strains of C. neoformans to wild-type genotype and phenotype, including prototrophy and growth rate. Furthermore, the unisexual cycle allows attenuated strains to purge deleterious mutations and produce progeny that are returned to wild-type virulence. Our results show that unisexual populations of C. neoformans are able to avoid Muller’s ratchet and loss of fitness through a unisexual reproduction cycle involving α-α cell fusion, nuclear fusion, and meiosis. Similar types of unisexual reproduction may operate in other pathogenic and saprobic eukaryotic taxa.