Studies of Enzyme Polymorphisms in the Kamuela Population of D. MERCATORUM. I. Estimation of the Level of Polymorphism

A Kamuela, Hawaii, population of Drosophila mercatorum was surveyed for enzyme variability. The mean heterozygosity and the proportion of polymorphic loci were estimated as 0.1255 and 0.37, respectively. Neither deviates more than one standard error from their respective means for 43 Drosophila species (Nevo 1978). Heterozygosity was distributed across enzyme categories in much the same manner as observed in other species (Gillespie and Kojima 1968; Johnson 1974), and enzymes associated with glycolysis were about as variable as other enzymes of central metabolism.--The levels of heterozygosity and polymorphism in this population do not seem to have been affected by a low-level capacity for parthenogenesis. The observed parthenogenetic reproduction is not strongly associated with particular allelic variants among viable parthenogenetic adults; however, the capacity to establish a self-sustaining parthenogenetic clone is strongly associated with the phenotype with the most frequent allele at every locus studied. We interpret these results to mean that isozyme variants do not strongly influence viability under total homozygosity (the genetic condition imposed by parthenogenesis), but they do have an impact upon the reproductive biology of parthenogenetic adults.     

Intra- and interspecific polymorphism of (AAT) n in microsatellite locus Du47D in parthenogenetic species of the genus Darevskia

The molecular structure of the allelic variants of (AAT) _n of the Du47D microsatellite locus was determined in parthenogenetic lizards Darevskia dahli, D. armeniaca, and D. rostombekovi. Comparative analysis of these alleles showed that they were characterized by perfect structure of microsatellite cluster, and were different in the number of (AAT) monomeric units, as well as in the combinations of species-specific substitutions and deletions in the microsatellite flanking regions. Molecular structure of microsatellite cluster, species-specific single nucleotide polymorphism (SNP), and different representation of alleles Du47 in the samples of parthenogenetic species examined point to the origin of the alleles from different bisexual species, which is consistent with the hybrid nature of unisexual species of the genus Darevskia. In addition, these data reflect different combination patterns of interspecific hybridization events with the participation of the same bisexual species upon the formation of hybrid genomes of parthenogenetic species. Possible application of the allelic variants of microsatellite loci of parthenogenetic lizards as the genetic markers for the analysis of the genomes of parthenogenetic species in the light of evolution, ecology, and parthenogenetic type of reproduction in vertebrates is discussed.     

Superinstability (TCT/TCC)n of microsatellite DNA in parthenogenetic lizards Darevskia unisexualis (family Lacertidae)

Using multilocus DNA fingerprinting, we have examined variability of (TCT)n microsatellite and M13 minisatellite DNA repeats in populations, families, and tissues of Caucasian parthenogenetic rock lizards Darevskia unisexualis (Lacertidae). It has been shown for the first time that population and family DNA samples of D. unisexualis (75 samples in total) have individually specific DNA fingerprinting patterns of (TCT)n fragments. Analysis of inheritance of (TCT)n microsatellites in 46 first-generation progeny in 17 parthenogenetic D. unisexualis families revealed their extremely high instability. Mutant TCT fingerprint phenotypes were found in virtually each animal of the progeny. Moreover, varying fragments in the progeny and their original variants in the mothers were shown to simultaneously contain (TCT)n and (TCC)n polypyrimidine clusters. At the same time, no variability of (TCT)n fragments has been detected in the tissues and organs of mature parthenogenetic lizards and in the analogous tissues of the two-week-old progeny of this year. This suggests the absence of somatic mosaicism and methylation of the corresponding loci in the samples. Along with the hyperinstability of (TCT/TCC)n polypyrimidine clusters, we have shown that the population and family DNA fingerprinting patterns of M13 minisatellites were invariable and monomorphic in the same DNA samples of D. unisexualis. Our results indicate that mutations at loci containing polypyrimidine microsatellites significantly contribute to the total genomic variability of parthenogenetic lizards D. unisexualis.     

Molecular characterization of allelic variants of (GATA)n microsatellite loci in parthenogenetic lizards Darevskia unisexualis (Lacertidae)

Populations of parthenogenetic lizards of the genus Darevskia consist of genetically identical animals, and represent a unique model for studying the molecular mechanisms underlying the variability and evolution of hypervariable DNA repeats. As unisexual lineages, parthenogenetic lizards are characterized by some level of genetic diversity at microsatellite loci. We cloned and sequenced a number of (GATA)n microsatellite loci of Darevskia unisexualis. PCR products from these loci were also sequenced and the degree of intraspecific polymorphism was assessed. Among the five (GATA)n loci analysed, two (Du215 and Du281) were polymorphic. Cross-species analysis of Du215 and Du281 indicate that the priming sites at the D. unisexualis loci are conserved in the bisexual parental species, D. raddei and D. valentini. Sequencing the PCR products amplified from Du215 and Du281 and from monomorphic Du323 showed that allelic differences at the polymorphic loci are caused by microsatellite mutations and by point mutations in the flanking regions. The haplotypes identified among the allelic variants of Du281 and among its orthologues in the parental species provide new evidence of the cross-species origin of D. unisexualis. To our knowledge, these data are the first to characterize the nucleotide sequences of allelic variants at microsatellite loci within parthenogenetic vertebrate animals.     

Low, but Strongly Structured Mitochondrial DNA Diversity in Root Knot Nematodes (Meloidogyne)

Root-knot nematodes (genus Meloidogyne) have been the subject of recent and numerous studies of genetic variation because of the need to develop molecular diagnostics for the four globally distributed, parthenogenetic species that are significant agricultural pests. Our analysis of Meloidogyne mtDNA improves on previous studies: (i) by examining restriction site polymorphism among a large number of isolates also characterized for standard morphological, host range and allozyme phenotypes; (ii) by using higher resolution electrophoretic techniques; and (iii) by mapping variable restriction sites with reference to the complete nucleotide sequence. This revealed fivefold less sequence divergence (<0.6%) between variants than estimated in previous restriction fragment length polymorphism (RFLP) studies, but perfect correspondence between mtDNA haplotype and allozyme (esterase) phenotypes. The mtDNA variation, although limited, is strongly structured with as much divergence between two lineages of Meloidogyne arenaria as between either of these and Meloidogyne javanica. The low diversity of mtDNAs suggests that these parthenogenetic lineages arose from distinct but closely related sexual females, a pattern seen in other parthenogenetic complexes. In contrast to the concordance between mtDNA and allozyme markers, there were several discrepancies between the traditional methods of identification. We suggest that further studies of these nematodes should focus on well defined genetic groups, whether or not these coincide with existing taxonomic units.     

Stability and variation in aphid clonal lineages

Evidence in the literature purporting to show genetic variability within parthenogenetic lines of aphids is reviewed. The results of three experimental approaches to this problem are reported: (1) a long term study of fluctuation in alate morph production in six lines of Myzus persicae; (2) an attempt tp select for caudal hair form in Acyrthosiphon pisum; (3) a study of the inheritance of esterase variants of M. persicae in sexual and parthenogenetic reproduction. It is concluded that genetic recombination during parthenogenesis is not the general rule in diese aphids, and that aphid parthenogenesis should be regarded as of ameiotic or apomictic type. Possible alternative explanations for the different kinds of variation or apparent variation which occur within aphid clonal lineages are discussed.     

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.     

Genetic polymorphism and evolution in parthenogenetic animals. I. Polyploid curculionidae

The genetic variability at enzyme loci in different triploid and tetraploid parthenogenetic weevil populations has been elucidated by starch gel electrophoresis. The overall genotype of individual weevils belonging to different populations has been determined for over 25 loci. The results are compared with those obtained for diploid bisexual races of either the same or closely related species. The variation within a parthenogenetic population differs from that in diploid, sexually reproducing populations, i.e. the allele frequencies are not in a Hardy-Weinberg equilibrium. The results indicate that apomictic parthenogenetic populations can differentiate genetically. The genotypes within a population resemble each other more than genotypes belonging to different populations. It is evident that evolution still continues—even if slowed down—in parthenogenetic weevils. A comparison between the allele relationships in geographically isolated polyploid parthenogenetic populations and related diploid bisexual forms does not support the hypothetical hybrid origin of parthenogenesis and polyploidy in weevils. Parthenogenesis within a parthenogenetic weevil species is evidently monophyletic.     

Molecular structure of the allelic variants of (AAT)n microsatellite locus Du47D in the parthenogenetic species Darevskia unisexualis and bisexual parental species D. valentini and D. raddei

Microsatellite repeats are one of the most widespread elements of the eukaryotic genome, but are poorly studied in species with clonal reproduction. PCR analysis and DNA sequencing were used to study the molecular structure of the allelic variants of microsatellite locus Du47D in the parthenogenetic species Darevskia unisexualis and its evolutionary ancestors, bisexual species D. raddei and D. valentini, of the genus Darevskia (Lacerta saxicola complex). Sequencing showed that the allelic variants of the D. unisexualis Du47D locus and the alleles of its D. raddei and D. valentini orthologs have a perfect microsatellite cluster structure, differ in number of ATT monomeric units, and have certain species-specific combinations of nucleotide substitutions, deletions, and insertions in the microsatellite-flanking DNA sequences. The Du47D alleles that the parthenogenetic species inherited from D. valentini or from D. raddei were identified.     

Developmental Potential of Haploid-derived Parthenogenetic Cells in Mouse Chimeric Embryos1

Studies were made on the contribution of haploid-derived parthenogenetic cells to haploid parthenogenetic ? fertilized chimeric embryos on day 9 and 10 of pregnancy. In most cases, the contribution of haploid-derived parthenogenetic cells to embryonic tissues was higher than that to extraembryonic tissues. The contribution of haploid-derived cells to embryonic tissues of some chimeras was more than 90%. Chromosomal analysis showed that actively dividing cells in most chimeric embryos contained about 40 chromosomes, indicating that they were diploidized, as haploid parthenogenetic blastocysts have about 20 chromosomes. Results suggested that haploid-derived parthehogenetic cells in chimeric embryos diploidized spontaneously after the blastocyst stage. These cells were capable of differentiating into most cell types of embryonic tissues, but scarcely differentiated into extraembryonic tissues of day 9 embryos. The fate of haploid-derived parthenogenetic cells during postimplantational development was similar to that of diploid parthenogenetic cells that had been diploidized experimentally in the one-cell stage.     

Molecular and genetic characterization of the allelic variants of Du215, Du281, Du323, and Du47G microsatellite loci in parthenogenetic lizard <Emphasis Type="Italic">Darevskia armeniaca</Emphasis> (Lacertidae)

A key issue in the study of unisexual (parthenogenetic) vertebrate species is the determination of their genetic and clonal diversity. In pursuing this aim, various markers of nuclear and mitochondrial genomes can be used. The most effective genetic markers include microsatellite DNA, characterized by high variability. The development and characterization of such markers is a necessary step in the genetic studies of parthenogenetic species. In the present study, using locus-specific PCR, for the first time, an analysis of allelic polymorphism of four microsatellite loci is performed in the populations of parthenogenetic species Darevskia armeniaca. In the studied populations, allelic variants of each locus are identified, and the nucleotide sequences of each allele are determined. It is demonstrated that allele differences are associated with the variation in the structure of microsatellite clusters and single nucleotide substitutions at fixed distances in flanking DNA regions. Structural allele variations form haplotype markers that are specific to each allele and are inherited from their parental bisexual species. It is established which of the parental alleles of each locus were inherited by the parthenogenetic species. The characteristics of the distribution and frequency of the alleles of microsatellite loci in the populations of D. armeniaca determining specific features of each population are obtained. The observed heterozygosity of the populations at the studied loci and the mutation rates in genome regions, as well as Nei’s genetic distances between the studied populations, are determined, and the phylogenetic relationships between them are established.     

PHYLOGENY AND EVOLUTION OF PARTHENOGENETIC WEEVILS OF THE ARAMIGUS TESSELLATUS SPECIES COMPLEX (COLEOPTERA: CURCULIONIDAE: NAUPACTINI): EVIDENCE FROM MITOCHONDRIAL DNA SEQUENCES

Molecular-phylogenetic studies of parthenogenetic animals have been a valuable recent addition to the literature on the evolutionary biology of sex. By illuminating the origins and ages of parthenogenetic lineages, such studies can help to define the temporal scale at which selection acts against parthenogenetic lineages, as well as provide an essential framework for further study. Although parthenogenetic weevils have played an important role in cytogenetic and protein-electrophoretic studies of parthenogenesis, they have not previously been subjects of DNA-based molecular-phylogenetic study. A mitochondrial DNA study of Aramigus tessellatus, a species complex of weevils native to South America, indentified 12 distinct (1–9% divergent) maternal lineages, of which 2 represent sexual populations, while at least 9 represent parthenogenetic lineages. These lineages partially correspond to lineages previously recognized by morphological differences. Phylogenetic analysis found 14 most parsimonious trees, according to which parthenogenesis appears to have arisen 3–7 times. There is a monophyletic group of lineages (the “brown clade”), having up to 4.5% sequence divergence within it, which may be primitively parthenogenetic and over 2 million years old.     

Life cycle of the cereal aphid Sitobion avenae F.: polymorphism and comparison of life history traits associated with sexuality

1. Clones of Sitobion avenae from the west of France exhibited four types of responses in growth chambers to short days and low temperatures: in addition to permanently parthenogenetic clones (anholocyclic) and clones producing both males and parthenogenetic females (androcyclic), lineages producing both mating females and males were categorized into holocyclic and intermediate types, the latter investing into both sexual and parthenogenetic reproduction.
2. Holocyclic, intermediate and androcyclic clones were tested outdoors for sexual production in autumn and results were consistent with those obtained in growth chambers. Eggs from holocyclic and intermediate clone selfings were laid and hatched for most crosses.
3. Reciprocal crosses within and between life cycle variants showed no effect of the type of cross on the number of eggs laid per mating female, their hatching success, or the survival and fecundity of the parthenogenetic females born from eggs (fundatrices).
4. Eggs from intermediate × intermediate crosses hatched 20 days earlier than those involving at least one holocyclic parent. It is suggested that this short diapausing time gives a selective advantage to intermediate clones in oceanic climates, which could compensate for their lower sexual production in autumn.
5. Selfed offspring showed inbreeding depression, which seems to be common in non host-alternating aphids.
6. Mating with androcyclic males affects percentage (lower) and time (longer) of egg hatching, but not the other investigated traits.
7. The significance of intragenotypic strategy mixing in androcyclic and intermediate clones is discussed as an overall adaptation to the uncertainty of winter climate in oceanic zones.     

Could sex be maintained through harmful males?

All else being equal, parthenogenetic females should produce as many surviving daughters as sexual couples produce daughters plus sons. Hence the resources spent on producing sons are a cost of sex and parthenogenetic females economize on sons. It has recently been shown that a small competitive advantage of sexual individuals can recoup this large reproductive disadvantage, while the adaptation behind the competitive advantage might differ from case to case. One hypothesis that has not yet been considered as a potential competitive advantage is that males could differentially harm parthenogenetic females, for example, through harassment, toxic seminal fluids, or infanticide. Harmful male functions result from the selection for males that maximise their fitness at the expense of females in the context of sexual conflict. Unless parthenogenetic lineages can maintain their resistance against harmful male functions, a competitive advantage for sex should be a by-product benefit of sexual conflict.
Mutations that make males harm parthenogenetic females worse than sexual ones, however, can be seen as evolutionary spite. The spiteful trait is not the production of costly sons, but the production of males that discriminate against parthenogenetic females. Spiteful behaviour can be positively selected, if it acts against negatively related victims. Sexual and parthenogenetic individuals within a population should usually be negatively related, because the genomes of the sexual individuals are bound together by recombination while those of parthenogenetic individuals will be identical except for divergence through mutation.
Some unusual cases of parthenogenesis are discussed in the light of this new hypothesis and an experimental approach for testing it is suggested.     

Moderate expression of Wnt signaling genes is essential for porcine parthenogenetic embryo development

Parthenogenetic embryos are invariably lost in mid-gestation, possibly due to the lack of the paternal genome and the consequent induction of aberrant gene expression. Wnt signaling is essential for embryonic development; however, the studies of this pathway in porcine parthenogenetic embryos have been limited. Here, the role of Wnt signaling in porcine parthenogenetic embryos was studied. In vivo embryos were used as controls. Single cell quantitative real-time PCR showed that Wnt signaling was down-regulated in porcine parthenogenetic embryos. Furthermore, immunofluorescence staining and real-time PCR demonstrated that porcine parthenogenetic embryo development was largely unaffected by the inhibition of Wnt signaling with IWP-2, but blastocyst hatching and trophectoderm development was blocked. In addition, parthenogenetic blastocyst hatching was improved by the activation of Wnt signaling by BIO. However, the developmental competency of porcine embryos, including blastocyst hatching, was impaired and apoptosis was induced upon the excessive activation of Wnt signaling. These findings constitute novel evidence that Wnt signaling is important for porcine pre-implantation development and that its down-regulation may lead to the low hatching rate of porcine parthenogenetic blastocysts.     

Hyperunstable (TCT/TCC) n Microsatellite Loci in Parthenogenetic Lizards Darevskia unisexualis (Lacertidae)

Using multilocus DNA fingerprinting, we have examined variability of (TCT) _n microsatellite and M13 minisatellite DNA repeats in populations, families, and tissues of parthenogenetic Caucasian rock lizards Darevskia unisexualis (Lacertidae). It has been shown for the first time that population and family DNA samples of D. unisexualis (75 samples in total) have individually specific DNA fingerprinting patterns of (TCT) _n fragments. Analysis of inheritance of (TCT) _n microsatellites in 46 first-generation progeny in 17 parthenogenetic D. unisexualis families revealed their extremely high instability. Mutant TCT fingerprint phenotypes were found in virtually each animal of the progeny. Moreover, varying fragments in the progeny and their original variants in the mothers were shown to simultaneously contain (TCT) _n and (TCC) _n polypyrimidine clusters. At the same time, no variability of (TCT) _n fragments has been detected in the tissues and organs of mature parthenogenetic lizards and in the analogous tissues of the two-week-old progeny of this year. This suggests the absence of somatic mosaicism and methylation of the corresponding loci in the samples. Along with the hyperinstability of (TCT/TCC) _n polypyrimidine clusters, we have shown that the population and family DNA fingerprinting patterns of M13 minisatellites were invariable and monomorphic in the same DNA samples of D. unisexualis.Our results indicate that mutations at loci containing polypyrimidine microsatellites significantly contribute to the total genomic variability of parthenogenetic lizards D. unisexualis.     

Genetic Polymorphism and Evolution in Parthenogenetic Animals. II. Diploid and Polyploid SOLENOBIA TRIQUETRELLA (Lepidoptera: Psychidae)

Genic polymorphism at sixteen enzyme loci of four different chromosomal races of Solenobia triquetrella (bisexual, two diploid parthenogenetic races and tetraploid parthenogenetic) has been studied by starch gel electrophoresis. Isolated small diploid bisexual populations have rather uniform allele frequencies at all loci which we have studied. Diploid and tetraploid parthenogenetic individuals of this species are in general as heterozygous as bisexual ones. All parthenogenetic local populations are different from each other in the Alps. These parthenogenetic genotypes cannot be derived from a common ancestor through single mutations but rather bear evidence for a polyphyletic origin of parthenogenesis in Solenobia triquetrella. In the marginal distribution areas of the species in northern Europe single genotypes are spread over far larger areas than in the mountain regions of central Europe. This may be due to the old origin of parthenogenesis and polyploidy in northern Europe. No new parthenogenetic and polyploid strains have lately arisen in the regions outside of the Alps.     

The activation of bovine oocytes to parthenogenetic development by ethanol

Dependence of parthenogenetic development ability of in vitro matured cow oocytes from their "aging" had been studied. It had been showed the possibility of parthenogenetic development of physiologically matured cow oocytes after inhibition of the first meiotic division and treatment with ethanol at least to 8-16-cell stage embryos.     

The histochemical identification of primordial germ cells in diploid parthenogenetic mouse embryos

An attempt has been made to improve the early post-implantation development potential of diploid parthenogenetic mouse embryos by transferring parthenogenetic blastocysts to one uterine horn of a pseudopregnant recipient and a similar number of fertilized embryos to the contralateral horn. In control studies, diploid parthenogenetic embryos were transferred to both uterine horns of appropriate recipients. Unfortunately no obvious advantage appeared to be gained by carrying out the former manoeuvre. A significant improvement in the development potential of the parthenogenones could have indicated that their poor post-implantation survival might have been associated with a deficiency, possibly of hormonal origin, in the functioning of their decidual reaction. However, sufficient somite-containing parthenogenetic embryos were obtained in this study to allow a comparison to be made between them and fertilized embryos that were morphologically at a comparable stage of development. The parthenogenones were found to have a markedly smaller crown-rump length than their fertilized counterparts. A high proportion of both the parthenogenetic and fertilized embryos were subsequently fixed and appropriately stained in order to localize alkaline phosphatase activity. The analysis of this material clearly demonstrated that parthenogenetic mouse embryos are in fact capable of producing primordial germ cells. The latter were recognized by their morphology, histochemical staining appearance, and characteristic location, being found in the early 'turned' embryos within the dorsal mesentery in close proximity to the developing gut tube, and in the more advanced limb-bud stage embryos within the gonadal ridges.     

Low parasitism rates in parthenogenetic bagworm moths do not support the parasitoid hypothesis for sex

The parasite hypothesis for sex is one of the many theories that have been suggested to solve the mystery of the widespread occurrence of sex despite its high short‐term costs. It suggests that sexual lineages have an evolutionary advantage over parthenogens because they can frequently generate new genotypes that are temporarily less prone to coevolving parasites. In this study, we looked for further supporting evidence for the parasite hypothesis of sex in an attempt to understand the coexistence of sexual and parthenogenetic bagworm moths (Naryciinae). The bagworm moths and their parasitoids form one of the few natural host–parasite systems where sexual and parthenogenetic hosts are apparently not separated by ecological or geographical barriers. Furthermore, in support of the parasite hypothesis for sex, parthenogenetic presence is negatively correlated with parasitism rate. We specifically tested, by identifying the reproductive mode of the parasitized individuals, whether parasitoids preferentially attack the parthenogens in sites with both sexual and parthenogenetic forms, as predicted by the parasite hypothesis. We collected hosts from sites with different frequencies of parthenogenetic and sexual moths. A DNA barcoding approach was used to determine the reproductive mode of the parasitized hosts. Furthermore, we investigated whether differences in host and parasitoid phenology could provide an alternative explanation for the variation in parasitism rates between parthenogens and sexuals. Our results contradict the prediction of the parasite hypothesis because parthenogenetic bagworm moths were less parasitized than sexuals in sympatric sites. Our findings can be explained by differences in phenology between the parthenogenetic and sexual moths rather than genetic incompatibility between parthenogenetic hosts and parasitoids. The stable coexistence of sexual and parthenogenetic Naryciinae despite the many apparent costs of sex in this system remains a mystery. Our work adds to the list of studies were the assumptions of the parasite hypothesis for sex are not all met.