Hybrid incompatibilities in the parasitic wasp genus Nasonia: negative effects of hemizygosity and the identification of transmission ratio distortion loci

The occurrence of hybrid incompatibilities forms an important stage during the evolution of reproductive isolation. In early stages of speciation, males and females often respond differently to hybridization. Haldane's rule states that the heterogametic sex suffers more from hybridization than the homogametic sex. Although haplodiploid reproduction (haploid males, diploid females) does not involve sex chromosomes, sex-specific incompatibilities are predicted to be prevalent in haplodiploid species. Here, we evaluate the effect of sex/ploidy level on hybrid incompatibilities and locate genomic regions that cause increased mortality rates in hybrid males of the haplodiploid wasps Nasonia vitripennis and Nasonia longicornis. Our data show that diploid F(1) hybrid females suffer less from hybridization than haploid F(2) hybrid males. The latter not only suffer from an increased mortality rate, but also from behavioural and spermatogenic sterility. Genetic mapping in recombinant F(2) male hybrids revealed that the observed hybrid mortality is most likely due to a disruption of cytonuclear interactions. As these sex-specific hybrid incompatibilities follow predictions based on Haldane's rule, our data accentuate the need to broaden the view of Haldane's rule to include species with haplodiploid sex determination, consistent with Haldane's original definition.     

Cytoplasmic Incompatibility and Bacterial Density in Nasonia Vitripennis

Cytoplasmically (maternally) inherited bacteria that cause reproductive incompatibility between strains are widespread among insects. In the parasitoid wasp Nasonia, incompatibility results in improper condensation and fragmentation of the paternal chromosomes in fertilized eggs. Some form of genome imprinting may be involved. Because of haplodiploidy, incompatibility results in conversion of (diploid) female eggs into (haploid) males. Experiments show that bacterial density is correlated with compatibility differences between male and female Nasonia. Males from strains with high bacterial numbers are incompatible with females from strains with lower numbers. Temporal changes in compatibility of females after tetracycline treatment are generally correlated with decreases in bacterial levels in eggs. However, complete loss of bacteria in mature eggs precedes conversion of eggs to the ``asymbiont' compatibility type by 3-4 days. This result is consistent with a critical ``imprinting' period during egg maturation, when cytoplasmic bacteria determine compatibility. Consequent inheritance of reduced bacterial numbers in F(1) progeny has different effects on compatibility type of subsequent male vs. female progeny. In some cases, partial incompatibility occurs which results in reduced offspring numbers, apparently due to incomplete paternal chromosome elimination resulting in aneuploidy.     

Evidence for female mortality in Wolbachia-mediated cytoplasmic incompatibility in haplodiploid insects: epidemiologic and evolutionary consequences

Abstract.— Until now, only two Wolbachia-mediated cytoplasmic incompatibility (CI) types have been described in haplodiploid species, the first in Nasonia (Insect) and the second in Tetranychus (Acari). They both induce a malebiased sex ratio in the incompatible cross. In Nasonia, CI does not reduce fertility since incompatible eggs develop as haploid males, whereas in Tetranychus CI leads to a partial mortality of incompatible eggs, thus reducing the fertility of females. Here, we study Wolbachia infection in a Drosophila parasitoid, Leptopilina heterotoma (Hymenoptera: Figitidae). A survey of Wolbachia infection shows that all natural populations tested are totally infected. Crosses between infected males and cured females show complete incompatibility: almost no females are produced. Moreover, incompatible eggs die early during their development, unlike Nasonia. This early death allows the parasitized Drosophila larva to achieve its development and to emerge. Thus, uninfected females crossed with infected males have reduced offspring production consisting only of males. Evidence of this CI type in insects demonstrates that the difference in CI types of Nasonia and Tetranychus is not due to specific factors of insects or acari. Using theoretical models, we compare the invasion processes of different strategies of Wolbachia: CI in diploid species, the two CI types in haplodiploid species, and parthenogenesis (the classical effect in haplodiploid species). Models show that CI in haplodiploid species is less efficient than in diploid ones. However, the Leptopilina type is advantageous compared to the Nasonia type. Parthenogenesis may be more or less advantageous, depending on the infection cost and on the proportion of fertilized eggs. Finally, we can propose different processes of Wolbachia strategy evolution in haplodiploid species from Nasonia CI type to Leptopilina CI type or parthenogenesis.     

Effective population size in Hymenoptera with complementary sex determination

Complementary sex determination in the haplodiploid Hymenoptera leads to the production of inviable or effectively sterile diploid males when diploid progeny are homozygous at the sex-determining locus. The production of diploid males reduces the number of females in a population and biases the effective breeding sex ratio in favor of haploid males. This in turn will reduce the effective population size (Ne) of hymenopteran populations with complementary sex determination relative to the expected reductions due to haplodiploidy alone. The effects of diploid male production on Ne in hymenopterans with complementary sex determination when diploid males are either inviable or effectively sterile are assessed theoretically. In both models, low allelic diversity at the sex locus reduces the Ne of hymenopteran populations, and this effect is largest when diploid males are effectively sterile.     

Phenotypic fitness effects of the selfish B chromosome,paternal sex ratio (PSR) in the parasitic waspNasonia vitripennis

Summary B chromosomes are often considered genomic parasites. Paternal sex ratio (PSR) is an extreme example of a parasitic B chromosome in the parasitoid waspNasonia vitripennis. PSR is transmitted through the sperm of carrier males and destroys the other paternal chromosomes in early fertilized eggs. PSR disrupts the normal haplodiploid sex determination in this wasp by converting diploid (female) eggs into haploid (male) eggs that bear PSR. In this study I compare a number of phenotypic fitness aspects of PSR and standard (non-PSR) males. In general, PSR males were as fit as standard males. No significant differences were found in longevity (with one exception), ability to compete for mates and sperm depletion rates. PSR males produced 11–22% larger family sizes and developed slightly faster than standard males. Under conditions of sperm competition, females who mated with both types of males fertilized a constant proportion of eggs with each sperm type over their lifetime. PSR males produced fewer offspring among progenies from double-inseminated females. Phenotypic fitness effects are believed to play a minor role in determining PSR frequencies in natural populations.     

Genetics of sex determination in the haplodiploid wasp <Emphasis Type="Italic">Nasonia vitripennis</Emphasis> (Hymenoptera: Chalcidoidea)

The parasitoid wasp Nasonia vitripennis reproduces by haplodiploidy; males are haploid and females are diploid. Sex determination in Nasonia is not governed by complementary alleles at one or more sex loci. As in most other insects, the sex-determining pathway consists of the basal switch doublesex that is sex-specifically regulated by transformer. Analysis of a polyploid and a mutant gynandromorphic strain, suggested a parent-specific effect (imprinting) on sex determination in Nasonia. Zygotic activity of transformer is autoregulated and depends on a combination of maternal provision of tra mRNA and a paternal genome set. This constitutes a novel way of transformer control in insect sex determination implying maternal imprinting. The nature of the maternal imprint is not yet known and it remains to be determined how broadly the Nasonia sex-determining mechanism applies to other haplodiploids.     

No patrigenes required for femaleness in the haplodiploid wasp Nasonia vitripennis

The parasitoid wasp Nasonia vitripennis is an emerging model organism for developmental and behavioral genetics. It reproduces by haplodiploidy; males typically develop parthenogenetically from haploid eggs and females from fertilized diploid eggs. A polyploid mutant strain is available in which females are triploid and lay haploid and diploid eggs that normally develop into males when unfertilized. In contrast to previous reports, approximately 2% of triploid females were found to occasionally produce daughters as well as gynandromorphs from diploid unfertilized eggs. Daughter production increased with age and differed among familial lineages. This is the first report of parthenogenetic female development in Nasonia. The results show that a paternally provided genome is not required for femaleness and call for modifications of existing models of sex determination in Nasonia.     

What haplodiploids can teach us about hybridization and speciation

Most evolutionary theory focuses on species that reproduce through sexual reproduction where both sexes have a diploid chromosome count. Yet a substantial proportion of multicellular species display complex life cycles, with both haploid and diploid life stages. A classic example is haplodiploidy, where females develop from fertilized eggs and are diploid, while males develop from unfertilized eggs and are haploid. Although haplodiploids make up about 15% of all animals (de la Filia et al. 2015 ), this type of reproduction is rarely considered in evolutionary theory. In this issue of Molecular Ecology, Patten et al. ( 2015 ) develop a theoretical model to compare the rate of nuclear and mitochondrial introgression in haplodiploid and diploid species. They show that when two haplodiploid species hybridize, nuclear genes are much less likely to cross the species barrier than if both species were to be diploids. The reason for this is that only half of the offspring resulting from matings between haplodiploid species are true hybrids: sons from such mating only inherit their mother genes and therefore only contain genes of the maternal species. Truly, hybrid males can only occur through backcrossing of a hybrid female to a male of one of the parental species. While this twist of haplodiploid transmission genetics limits nuclear introgression, mitochondrial genes, which are maternally inherited, are unaffected by the scarcity of hybrid males. In other words, the rate of mitochondrial introgression is the same for haplodiploid and diploid species. As a result, haplodiploid species on average show a bias of mitochondrial compared to nuclear introgression.     

PSR (paternal sex ratio) chromosomes: the ultimate selfish genetic elements

PSR (paternal sex ratio) chromosomes are a type of supernumerary (or B) chromosomes that occur in haplodiploid arthropods. They are transmitted through sperm but then cause loss of the paternal chromosomes (except themselves) early in development. As a result, PSR chromosomes convert diploid fertilized eggs (which would normally develop into females) into haploid males that carry a PSR chromosome. Because they act by completely eliminating the haploid genome of their hosts, PSR chromosomes are the most extreme form of selfish or parasitic DNA known. PSR was originally described in the parasitic wasp Nasonia vitripennis (Pteromalidae). A second PSR chromosome has been found in Trichogramma kaykai, an egg parasitoid from a different family of Hymenoptera (Trichogrammatidae). We argue that PSR chromosomes are likely to be widespread in haplodiploid organisms, but have so far gone under reported due to a paucity of population genetic studies in haplodiploids. We describe the two known PSR systems and related phenomena, and models indicating the conditions conducive to increase of PSR like chromosomes in haplodiploids.     

The kin selection theory of genomic imprinting and modes of reproduction in the eusocial Hymenoptera

Genomic imprinting is known from flowering plants and mammals but has not been confirmed for the Hymenoptera even though the eusocial Hymenoptera are prime candidates for this peculiar form of gene expression. Here, the kin selection theory of genomic imprinting is reviewed and applied to the eusocial Hymenoptera. The evidence for imprinting in eusocial Hymenoptera with the typical mode of reproduction, involving the sexual production of diploid female offspring, which develop into workers or gynes, and the arrhenotokous parthenogenesis of haploid males, is also reviewed briefly. However, the focus of this review is how atypical modes of reproduction, involving thelytokous parthenogenesis, hybridisation and androgenesis, may also select for imprinting. In particular, naturally occurring hybridisation in several genera of ants may provide useful tests of the role of kin selection in the evolution of imprinting. Hybridisation is expected to disrupt the coadaptation of antagonistically imprinted loci, and thus affect the phenotypes of hybrids. Some of the limited data available on hybrid worker reproduction and on colony sex ratios support predictions about patterns of imprinting derived from kin selection theory.     

Epigenetic inheritance and genome regulation: is DNA methylation linked to ploidy in haplodiploid insects?

Organisms show great variation in ploidy level. For example, chromosome copy number varies among cells, individuals and species. One particularly widespread example of ploidy variation is found in haplodiploid taxa, wherein males are typically haploid and females are typically diploid. Despite the prevalence of haplodiploidy, the regulatory consequences of having separate haploid and diploid genomes are poorly understood. In particular, it remains unknown whether epigenetic mechanisms contribute to regulatory compensation for genome dosage. To gain greater insights into the importance of epigenetic information to ploidy compensation, we examined DNA methylation differences among diploid queen, diploid worker, haploid male and diploid male Solenopsis invicta fire ants. Surprisingly, we found that morphologically dissimilar diploid males, queens and workers were more similar to one another in terms of DNA methylation than were morphologically similar haploid and diploid males. Moreover, methylation level was positively associated with gene expression for genes that were differentially methylated in haploid and diploid castes. These data demonstrate that intragenic DNA methylation levels differ among individuals of distinct ploidy and are positively associated with levels of gene expression. Thus, these results suggest that epigenetic information may be linked to ploidy compensation in haplodiploid insects. Overall, this study suggests that epigenetic mechanisms may be important to maintaining appropriate patterns of gene regulation in biological systems that differ in genome copy number.     

Deletion Analysis of the Selfish B Chromosome, Paternal Sex Ratio (Psr), in the Parasitic Wasp Nasonia Vitripennis

Paternal Sex Ratio (PSR) is a ``selfish' B chromosome in the parasitoid wasp Nasonia vitripennis. It is transmitted via sperm, but causes supercondensation and destruction of the paternal chromosomes in early fertilized eggs. Because this wasp has haplodiploid sex determination, the effect of PSR is to convert diploid (female) eggs into haploid (male) eggs that carry PSR. Characterizing its genetic structure is a first step toward understanding mechanisms of PSR action. The chromosome is largely heterochromatic and contains several tandemly repeated DNA sequences that are not present on the autosomes. A deletion analysis of PSR was performed to investigate organization of repeats and location of functional domains causing paternal chromosome destruction. Deletion profiles using probes to PSR-specific repetitive DNA indicate that most repeats are organized in blocks on the chromosome. This study shows that the functional domains of PSR can be deleted, resulting in nonfunctional PSR chromosomes that are transmitted to daughters. A functional domain may be linked with the psr22 repeat, but function may also depend on abundance of PSR-specific repeats on the chromosome. It is hypothesized that the repeats act as a ``sink' for a product required for proper paternal chromosome processing. Almost all deletion chromosomes remained either functional of nonfunctional in subsequent generations following their creation. One chromosome was exceptional in that it reverted from nonfunctionality to functionality in one lineage. Transmission rates of nonfunctional deletion chromosomes were high through haploid males, but low through diploid females.     

Haplodiploidy, sex, and the evolution of pesticide resistance

The assumption that males and females are equally tolerant to pesticides in haplodiploid arthropods led to the prediction that the evolution of resistance is faster in haplodiploid than in diploid arthropods. However, in this review, it was found that the ratio of male to female tolerance is substantially smaller in haplodiploid than in diploid arthropods, indicating that resistance alleles are not strongly up-regulated in haploid males. In addition, males were generally less tolerant than females in both haplodiploid and diploid arthropods. Factors such as sexual size dimorphism and sex-dependent selection may account for the lower tolerance in males than in females. Little among-population variation in the ratio of male to female tolerance was found in three species. Moreover, the tolerance ratio generally remained unchanged by selection for resistance to pesticides, although significant among-species variation was present within arthropod orders. This indicates that sexual dimorphism in pesticide tolerance evolves at a slower rate than resistance to pesticides. Simulations considering between-sex differences in pesticide tolerance showed that resistance evolution can be slower in haplodiploids than in diploids. Recessive resistance, low male tolerance to pesticides, fitness costs expressed in males, and the use of refuges contributed in substantially delaying the evolution of resistance in haplodiploid arthropods. These findings cast a new perspective on the evolution of pesticide resistance in haplodiploid herbivores and natural enemies.     

Sex determination in the haplodiploid wasp Nasonia vitripennis (Hymenoptera: Chalcidoidea): a critical consideration of models and evidence

Sex determining mechanisms are highly diverse. Like all Hymenoptera, the parasitic wasp Nasonia vitripennis reproduces by haplodiploidy: males are haploid and females are diploid. Sex in Nasonia is not determined by complementary alleles at sex loci. Evidence for several alternative models is considered. Recent studies on a polyploid and a gynandromorphic mutant strain point to a maternal product that is balanced against the number of chromosomal complements in the zygote and a parent-specific (imprinting) effect. Research is now focused on the molecular details of sex determination in Nasonia.     

Segregation and Recombination of Non-Mendelian Genes in Chlamydomonas

Non-Mendelian genes in Chamydomonas reinhardtii are inherited in a uniparental (UP) fashion. Most zygotes and their progeny receive UP genes only from the mt(+) or maternal parent. However, a few exceptional zygotes are also found in which the mt(-) or paternal UP genome is transmitted. Most of the exceptional zygotes are biparental in that their progeny segregate UP genes transmitted by both parents. As a result, biparental zygotes have been extensively used to study the rules governing UP inheritance.The frequency of biparental zygotes can be greatly increased if the maternal parent is irradiated with ultraviolet light prior to mating. Based principally on studies with ultraviolet-induced biparental zygotes, Sager has argued that a vegetative cell contains two copies of the UP genome and that the progeny of a biparental zygote receive a copy derived from each parent. Results reported in this paper with spontaneous and ultraviolet-induced biparental zygotes do not support the two copy model, but argue for a mulitple copy model with most of the copies normally being transmitted by the maternal parent. A multiple copy model which accounts for both Sager's results and ours is presented.     

The relationship between DNA methylation and chromosome imprinting in the coccid Planococcus citri

The phenomenon of chromosome, or genomic, imprinting indicates the relevance of parental origin in determining functional differences between alleles, homologous chromosomes, or haploid sets. In mealybug males (Homoptera, Coccoidea), the haploid set of paternal origin undergoes heterochromatization at midcleavage and remains so in most of the tissues. This different behavior of the two haploid sets, which depends on their parental origin, represents one of the most striking examples of chromosome imprinting. In mammals, DNA methylation has been postulated as a possible molecular mechanism to differentially imprint DNA sequences during spermatogenesis or oogenesis. In the present article we addressed the role of DNA methylation in the imprinting of whole haploid sets as it occurs in Coccids. We investigated the DNA methylation patterns at both the molecular and chromosomal level in the mealybug Planococcus citri. We found that in both males and females the paternally derived haploid set is hypomethylated with respect to the maternally derived one. Therefore, in males, it is the paternally derived hypomethylated haploid set that is heterochromatized. Our data suggest that the two haploid sets are imprinted by parent-of-origin-specific DNA methylation with no correlation with the known gene-silencing properties of this base modification.     

The dice of fate: the csd gene and how its allelic composition regulates sexual development in the honey bee, Apis mellifera

Perhaps 20% of known animal species are haplodiploid: unfertilized haploid eggs develop into males and fertilized diploid eggs into females. Sex determination in such haplodiploid species does not rely on a difference in heteromorphic sex chromosome composition but the genetic basis has been elucidated in some hymenopteran insects (wasps, sawflies, ants, bees). In these species, the development into one sex or the others depends on an initial signal whether there is only one allele or two different alleles of a single gene, the complementary sex determiner (csd), in the zygotic genome. The gene has been most-recently identified in the honey bee and has been found to encode an arginine serine-rich (SR) type protein. Heterozygosity generates an active protein that initiates female development while hemizygosity/homozygosity results in a non-active CSD protein and default male development. I will discuss plausible models of how the molecular decision of male and female is made and implemented. Comparison to hierarchies of dipteran insects suggests that SR-type protein has facilitated the differentiation of sex-determining systems and hierarchies.     

A genetic screen for zygotic embryonic lethal mutations affecting cuticular morphology in the wasp Nasonia vitripennis

We have screened for zygotic embryonic lethal mutations affecting cuticular morphology in Nasonia vitripennis (Hymenoptera; Chalcidoidea). Our broad goal was to investigate the use of Nasonia for genetically surveying conservation and change in regulatory gene systems, as a means to understand the diversity of developmental strategies that have arisen during the course of evolution. Specifically, we aim to compare anteroposterior patterning gene functions in two long germ band insects, Nasonia and Drosophila. In Nasonia, unfertilized eggs develop as haploid males while fertilized eggs develop as diploid females, so the entire genome can be screened for recessive zygotic mutations by examining the progeny of F1 females. We describe 74 of >100 lines with embryonic cuticular mutant phenotypes, including representatives of coordinate, gap, pair-rule, segment polarity, homeotic, and Polycomb group functions, as well as mutants with novel phenotypes not directly comparable to those of known Drosophila genes. We conclude that Nasonia is a tractable experimental organism for comparative developmental genetic study. The mutants isolated here have begun to outline the extent of conservation and change in the genetic programs controlling embryonic patterning in Nasonia and Drosophila.     

膜翅目昆虫的性别决定机制

昆虫性别决定机制存在多样性和复杂性,其中膜翅目昆虫的性别决定由单双倍体决定,单倍体为雄性,二倍体为雌性。本文就膜翅目昆虫的性别决定模式和分子机制进行综述。膜翅目昆虫性别决定有6种模式,即互补性性别决定(complementary sex determination, CSD)、多位点互补性性别决定(multiple-locus CSD, ml-CSD)、基因组印记、母体效应、内共生体诱导产雌单性生殖、父本遗传基因组消除(paternal genome elimination, PGE)。其中,CSD机制是目前在膜翅目昆虫中普遍接受的性别决定模式。而蜜蜂的CSD性别决定机制是膜翅目昆虫性别决定模式中的典型代表,受csd→fem→dsx这一调控级联的控制。     

The inheritance of organelle genes and genomes: patterns and mechanisms.

Unlike nuclear genes and genomes, the inheritance of organelle genes and genomes does not follow Mendel's laws. In this mini-review, I summarize recent research progress on the patterns and mechanisms of the inheritance of organelle genes and genomes. While most sexual eukaryotes show uniparental inheritance of organelle genes and genomes in some progeny at least part of the time, increasing evidence indicates that strictly uniparental inheritance is rare and that organelle inheritance patterns are very diverse and complex. In contrast with the predominance of uniparental inheritance in multicellular organisms, organelle genes in eukaryotic microorganisms, such as protists, algae, and fungi, typically show a greater diversity of inheritance patterns, with sex-determining loci playing significant roles. The diverse patterns of inheritance are matched by the rich variety of potential mechanisms. Indeed, many factors, both deterministic and stochastic, can influence observed patterns of organelle inheritance. Interestingly, in multicellular organisms, progeny from interspecific crosses seem to exhibit more frequent paternal leakage and biparental organelle genome inheritance than those from intraspecific crosses. The recent observation of a sex-determining gene in the basidiomycete yeast Cryptococcus neoformans, which controls mitochondrial DNA inheritance, has opened up potentially exciting research opportunities for identifying specific molecular genetic pathways that control organelle inheritance, as well as for testing evolutionary hypotheses regarding the prevalence of uniparental inheritance of organelle genes and genomes.