GENETIC ARCHITECTURE OF ISOLATION BETWEEN TWO SPECIES OF SILENE WITH SEX CHROMOSOMES AND HALDANE'S RULE

Examination of the genetic architecture of hybrid breakdown can provide insight into the genetic mechanisms of commonly observed isolating phenomena such as Haldane's rule. We used line‐cross analysis to dissect the genetic architecture of divergence between two plant species that exhibit Haldane's rule for male sterility and rarity, Silene latifolia and Silene diclinis. We made 15 types of crosses, including reciprocal F₁, F₂, backcrosses, and later‐generation crosses, grew the seeds to flowering, and measured the number of viable ovules, proportion of viable pollen, and sex ratio. Typically, Haldane's rule for male rarity in XY animal hybrids is explained by interactions involving recessive X‐linked alleles that are deleterious when hemizygous (dominance theory), whereas sterility is explained by rapid evolution of spermatogenesis genes (faster‐male evolution). In contrast, we found that the genetic mechanisms underlying Haldane's rule between the two Silene species did not follow these conventions. Dominance theory was sufficient to explain male sterility, but male rarity likely involved faster‐male evolution. We also found an effect of the neo‐sex chromosomes of S. diclinis on the extreme rarity of some hybrid males. Our findings suggest that the genetic architecture of Haldane's rule in dioecious plants may differ from those commonly found in animals.     

Differential strength of sex-biased hybrid inferiority in impeding gene flow may be a cause of Haldane's rule

Abstract In animals, if one sex of the F₁ hybrid between two species is sterile or inviable, it is usually the heterogametic (XY or WZ) sex. This phenomenon, known as Haldane's rule, is currently thought to be coincidentally caused by different mechanisms in separate entities. The following questions have never been asked: Are heterogametic and homogametic inferiority (sterility or inviability) equivalent as isolating mechanisms? Could discrepancies between them, if existing, produce Haldane's rule? Here I consider sex‐biased hybrid inferiority strictly as an isolating mechanism, and quantitatively evaluate its strength in impeding gene flow. The comparison reveals that the ability of sex‐biased inferiority to impede gene flow varies according to the sex and chromosome involved. Heterogametic inferiority is a weaker barrier when unidirectional and a much stronger one when in compound reciprocal directions, compared with homogametic inferiority. Such differential strength may affect divergence in speciation and produce Haldane's rule.     

TEMPERATURE EFFECTS AND GENOTYPE-BY-ENVIRONMENT INTERACTIONS IN HYBRIDS: HALDANE'S RULE IN FLOUR BEETLES

When males of the flour beetle, Tribolium castaneum, are crossed to females of its close relative T. freemani, the sex ratio of the hybrids is female biased, owing in part to hybrid male mortality. Morphological abnormalities are also frequent in the surviving hybrid males, but not in the hybrid females. The finding that the heterogametic sex (male) is more adversely affected in interspecific crosses than the homogametic sex is consistent with Haldane's rule, which predicts that hybrid dysfunction should emerge as an indirect byproduct of divergent adaptation to differing environments. If so, environmental effects and genotype-by-environment interactions (GEI) should characterize the expression of Haldane's rule and interspecific hybrid traits in general. We used two wild-collected populations of T. castaneum (from Infantes, Spain, and Madagascar) to investigate the effects of environmental variation on the expression of Haldane's rule. Males from each population were mated to several T. freemani females and the half-sibling hybrid progenies were reared across a series of temperature regimes. For both populations, we found that hybrids raised at higher temperatures exhibited a more extreme expression of Haldane's rule: The hybrid sex ratios were more biased toward females and hybrid males had a much higher incidence of morphological abnormalities. The average response to temperature, the norm of reaction for Haldane's rule, varied between the two populations, and we found considerable and significant GEI for both hybrid traits within both populations. The evolutionary implications of these findings are discussed in the context of speciation arising as an indirect effect of local adaptation.     

100 years of Haldane's rule

Haldane's rule is one of the ‘two rules of speciation’. It states that if one sex is ‘absent, rare or sterile’ in a hybrid population, then that sex will be heterogametic. Since Haldane first made this observation, 100 years have passed and still questions arise over how many independent examples exist and what the underlying causes of Haldane's rule are. This review aims to examine research that has occurred over the last century. It seeks to do so by discussing possible causes of Haldane's rule, as well as gaps in the research of these causes that could be readily addressed today. After 100 years of research, it can be concluded that Haldane's rule is a complicated one, and much current knowledge has been accrued by studying the model organisms of speciation. This has led to the primacy of dominance theory and faster-male theory as explanations for Haldane's rule. However, some of the most interesting findings of the 21st century with regard to Haldane's rule have involved investigating a wider range of taxa emphasizing the need to continue using comparative methods, including ever more taxa as new cases are discovered.     

X‐AUTOSOME INCOMPATIBILITIES IN DROSOPHILA MELANOGASTER: TESTS OF HALDANE'S RULE AND GEOGRAPHIC PATTERNS WITHIN SPECIES

Substantial genetic variation exists in natural populations of Drosophila melanogaster. This segregating variation includes alleles at different loci that interact to cause lethality or sterility (synthetic incompatibilities). Fitness epistasis in natural populations has important implications for speciation and the rate of adaptive evolution. To assess the prevalence of epistatic fitness interactions, we placed naturally occurring X chromosomes into genetic backgrounds derived from different geographic locations. Considerable amounts of synthetic incompatibilities were observed between X chromosomes and autosomes: greater than 44% of all combinations were either lethal or sterile. Sex‐specific lethality and sterility were also tested to determine whether Haldane's rule holds for within‐species variation. Surprisingly, we observed an excess of female sterility in genotypes that were homozygous, but not heterozygous, for the X chromosome. The recessive nature of these incompatibilities is similar to that predicted for incompatibilities underlying Haldane's rule. Our study also found higher levels of sterility and lethality for genomes that contain chromosomes from different geographical regions. These findings are consistent with the view that genomes are coadapted gene complexes and that geography affects the likelihood of epistatic fitness interactions.     

Biased introgression of mitochondrial and nuclear genes: a comparison of diploid and haplodiploid systems

Hybridization between recently diverged species, even if infrequent, can lead to the introgression of genes from one species into another. The rates of mitochondrial and nuclear introgression often differ, with some taxa showing biases for mitochondrial introgression and others for nuclear introgression. Several hypotheses exist to explain such biases, including adaptive introgression, sex differences in dispersal rates, sex‐specific prezygotic isolation and sex‐specific fitness of hybrids (e.g. Haldane's rule). We derive a simple population genetic model that permits an analysis of sex‐specific demographic and fitness parameters and measures the relative rates of mitochondrial and nuclear introgression between hybridizing pairs. We do this separately for diploid and haplodiploid species. For diploid taxa, we recover results consistent with previous hypotheses: an excess of one sex among the hybridizing migrants or sex‐specific prezygotic isolation causes a bias for one type of marker or the other; when Haldane's rule is obeyed, we find a mitochondrial bias in XY systems and a nuclear bias in ZW systems. For haplodiploid taxa, the model reveals that owing to their unique transmission genetics, they are seemingly assured of strong mitochondrial biases in introgression rates, unlike diploid taxa, where the relative fitness of male and female hybrids can tip the bias in either direction. This heretofore overlooked aspect of hybridization in haplodiploids provides what is perhaps the most likely explanation for differential introgression of mitochondrial and nuclear markers and raises concerns about the use of mitochondrial DNA barcodes for species delimitation in these taxa.     

Gene flow across a hybrid zone maintained by a weak heterogametic incompatibility and positive selection of incompatible alleles

Hybridization between incipient species is more likely to produce sterile or inviable F₁ offspring in the heterogametic (XY or ZW) sex than in the homogametic (XX or ZZ) sex, a phenomenon known as Haldane's rule. Population dynamics associated with Haldane's rule may play an important role in early speciation of sexually reproducing organisms. The dynamics of the hybrid zone maintained by incomplete hybrid inferiority (sterility/inviability) in the heterogametic sex (a ‘weak’ Haldane's rule) caused by a Bateson–Dobzhansky–Muller incompatibility was modelled. The influences and interplays of the strengths of incompatibility, dispersal, density‐dependent regulation (DDR) and local adaptation of incompatible alleles in a scenario of short‐range dispersal (the stepping‐stone model) were examined. It was found that a partial heterogametic hybrid incompatibility could efficiently impede gene flow and maintain characteristic clinal noncoincidence and discordance of alleles. Density‐dependent regulation appears to be an important factor affecting hybrid zone dynamics: it can effectively skew the effects of the partial incompatibility and dispersal as measured by effective dispersal, clinal structures and density depression. Unexpectedly, local adaptation of incompatible alleles in the parental populations, which would be critical for the establishment of the incompatibility, exerts little effect on hybrid zone dynamics. These results strongly support the plausibility of the adaptive origin of hybrid incompatibility and ecological speciation: an adaptive mutation, if it confers a marginal fitness advantage in the local population and happens to cause epistatic inferiority in hybrids, could efficiently drive further genetic divergence that may result in the gene becoming an evolutionary hotspot.     

Speciation in two neotropical butterflies: extending Haldane's rule

Anartia fatima and A. amathea form a hybrid zone in Panama where F1 and back-cross hybrids are found. Crosses were carried out to determine the nature of any reproductive isolation between these two butterflies. A novel analysis demonstrated both strong assortative mating among the pure forms and an unusual example of Haldane''s rule: F1 hybrid females (the heterogametic sex) from the cross A. amathea (female) multiplied by A. fatima (male) have a reduced tendency to mate. Historically, Haldane''s rule has been restricted to hybrid mortality or sterility and most studies have concentrated on taxa (predominantly Drosophila) between which strong barriers to gene flow already exist. Our data suggest that Haldane''s rule might be extended to cover any decrease in hybrid fitness and that mating propensity may provide a sensitive and comparable means of assessing such decreases. Other barriers to gene flow were also evident in Anartia: F1 hybrid females have reduced fertility (also a Haldane effect) and larval survivorship was greatly reduced in F2 hybrids of both sexes. These examples of hybrid disruption are expected under the dominance theory of Haldane''s rule but do not exclude other explanations.     

Increased differentiation and reduced gene flow in sex chromosomes relative to autosomes between lineages of the brown creeper Certhia americana

The properties of sex chromosomes, including patterns of inheritance, reduced levels of recombination, and hemizygosity in one of the sexes may result in the faster fixation of new mutations via drift and natural selection. Due to these patterns and processes, the two rules of speciation to describe the genetics of postzygotic isolation, Haldane's rule and the large‐X effect, both explicitly include quicker evolution on sex chromosomes relative to autosomes. Because sex‐linked mutations may be the first to become fixed in the speciation process, and appear to be due to stronger genetic drift (in birds), we may identify pronounced genetic differentiation in sex chromosomes in taxa experiencing recent speciation and diverging mainly via genetic drift. Here, we use nine sex‐linked and 21 autosomal genetic markers to investigate differential divergence and introgression between marker types in Certhia americana. We identified increased levels of genetic differentiation and reduced levels of gene flow on sex chromosomes relative to autosomes. This pattern is similar to those observed in other recently‐divergent avian species, providing another case study of the earlier role of sex chromosomes in divergence, relative to autosomes. Additionally, we identify three markers that may be under selection between Certhia americana lineages.     

Gametic selection,developmental trajectories,and extrinsic heterogeneity in Haldane's rule

Deciphering the genetic and developmental causes of the disproportionate rarity, inviability, and sterility of hybrid males, Haldane's rule, is important for understanding the evolution of reproductive isolation between species. Moreover, extrinsic and prezygotic factors can contribute to the magnitude of intrinsic isolation experienced between species with partial reproductive compatibility. Here, we use the nematodes Caenorhabditis briggsae and C. nigoni to quantify the sensitivity of hybrid male viability to extrinsic temperature and developmental timing, and test for a role of mito‐nuclear incompatibility as a genetic cause. We demonstrate that hybrid male inviability manifests almost entirely as embryonic, not larval, arrest and is maximal at the lowest rearing temperatures, indicating an intrinsic‐by‐extrinsic interaction to hybrid inviability. Crosses using mitochondrial substitution strains that have reciprocally introgressed mitochondrial and nuclear genomes show that mito‐nuclear incompatibility is not a dominant contributor to postzygotic isolation and does not drive Haldane's rule in this system. Crosses also reveal that competitive superiority of X‐bearing sperm provides a novel means by which postmating prezygotic factors exacerbate the rarity of hybrid males. These findings highlight the important roles of gametic, developmental, and extrinsic factors in modulating the manifestation of Haldane's rule.     

Evolution of ZZ/ZW and XX/XY sex-determination systems in the closely related medaka species, <Emphasis Type="Italic">Oryzias hubbsi</Emphasis> and <Emphasis Type="Italic">O. dancena</Emphasis>

A DM-domain gene on the Y chromosome was identified as the sex-determining gene in the medaka, Oryzias latipes, and named DMY (also known as dmrt1bY). However, this gene is absent in most Oryzias fishes, suggesting that closely related species have another sex-determining gene. In fact, it has been demonstrated that the Y chromosome in O. dancena is not homologous to that in O. latipes, whereas both species have an XX/XY sex-determination system. Through a progeny test of sex-reversed fish and a linkage analysis of isolated sex-linked DNA markers, we show that O. hubbsi, which is one of the most closely related species to O. dancena, has a ZZ/ZW system. In addition, genetic and fluorescence in situ hybridization mapping of the sex-linked markers revealed that sex chromosomes in O. hubbsi and O. dancena are not homologous, indicating different origins of these ZW and XY sex chromosomes. Furthermore, we found that O. hubbsi has morphologically heteromorphic sex chromosomes, in which the W chromosome has 4,6-diamidino-2-phenylindole (DAPI)-positive heterochromatin blocks and is larger than the Z chromosome, although such differentiated sex chromosomes have not been observed in other Oryzias species. These findings suggest that a variety of sex-determining mechanisms and sex chromosomes have evolved in Oryzias.     

Haldane rules: costs of outbreeding at production of daughters in sand lizards

Haldane's rule is one of the most widely applicable paradigms in evolutionary biology, stating that in species crossings, the heterogametic sex will suffer more severely in terms of sterility and inviability. We address this in a within‐species outbreeding situation by assessing the risk of producing inviable offspring depending on the sex ratio of the clutch produced in between‐population crossings in the laboratory. In crossings between male and female sand lizards (Lacerta agilis) from two different sampling regions, one in Sweden, one in central Europe, risk of gametic incompatibility is unaffected by outbreeding, but offspring from between‐population crossings show 300% higher malformation frequency and 10% lower hatching success. The risk of having inviable offspring increases with the production of daughters, i.e. the hemizygous sex in this species (ZW). Such sex‐specific genetic costs of offspring production need to be incorporated into life history ecology, e.g. sex allocation theory.     

DIVERGENCE OF MEIOTIC DRIVE-SUPPRESSION SYSTEMS AS AN EXPLANATION FOR SEX-BIASED HYBRID STERILITY AND INVIABILITY

Two empirical generalizations about speciation remain unexplained: the tendency of the heterogametic sex to be sterile or inviable in F₁ hybrids (Haldane's rule), and the tendency of the X chromosome to harbor the genetic elements that cause this sex bias in hybrid fitness. I suggest that divergence of meiotic drive systems on the sex chromosomes can explain these observations. The theory follows from two simple facts. First, sex chromosomes are particularly susceptible to the forces of meiotic drive. Second, divergence of meiotic drive systems can cause hybrid sterility and in viability. The main objection to the theory is that meiotic drive is apparently rare, whereas the observed pattern of hybrid fitness is widespread. I answer this objection by showing that divergence of meiotic drive systems can explain the two generalizations even if large departures from Mendelian segregation are rarely observed.     

HALDANE'S RULE AND SEX BIASSED GENE FLOW BETWEEN TWO HYBRIDIZING FLYCATCHER SPECIES (FICEDULA ALBICOLLIS AND F. HYPOLEUCA,AVES: MUSCICAPIDAE)

The collared flycatcher (Ficedula albicollis) and the pied flycatcher (F. hypoleuca) hybridize where their geographic ranges overlap. Restriction fragment comparison of 5% of the mitochondrial genome showed a sequence divergence of 10% between these flycatcher species. This degree of sequence divergence between a closely related pair of bird species is unusually high and contrasts with the low level of divergence between F. albicollis and F. hypoleuca in nuclear genes (Nei's D = 0.0006) revealed by enzyme electrophoresis. The low nuclear differentiation is explained by sex biassed gene flow and introgression in nuclear genes (via fertile male hybrids), while the high mitochondrial DNA sequence divergence is preserved by sterility of female hybrids, which prevents mitochondrial introgression. This pattern is in accordance with Haldane's rule and is supported by field data on hybrid fertility. The high mtDNA differentiation could be explained by transfer of mitochondrial DNA from a third species during a past period of hybridization.     

The genomics of incompatibility factors and sex determination in hybridizing species of Cottus (Pisces)

Cottus rhenanus and Cottus perifretum have formed hybrid lineages and narrow hybrid zones that can be best explained through the action of natural selection. However, the underlying selective forces as well as their genomic targets are not well understood. This study identifies genomic regions in the parental species that cause hybrid incompatibilities and tests whether these manifest in a sex-specific manner to learn about processes that affect natural hybridization in Cottus. Interspecific F₂ crosses were analyzed for 255 markers for genetic mapping and to detect transmission distortion as a sign for genetic incompatibilities. The Cottus map consists of 24 linkage groups with a total length of 1575.4 cM. A male heterogametic (XY) sex determination region was found on different linkage groups in the two parental species. Genetic incompatibilities were incomplete, varied among individuals and populations and were not associated with the heterogametic sex. The variance between populations and individuals makes it unlikely that there are species-specific incompatibility loci that could affect the gene pool of natural hybrids in a simple and predictable way. Conserved synteny with sequenced fish genomes permits to genetically study the Cottus genome through the transfer of genomic information from the model fish species. Homology relationships of candidate genomic regions in Cottus indicate that sex determination is not based on the same genomic regions found in other fish species. This suggests a fast evolutionary turnover of the genetic basis of sex determination that, together with the small size of the heterogametic regions, may contribute to the absence of fitness effects related to the Haldane''s rule.     

Whole chromosome painting reveals independent origin of sex chromosomes in closely related forms of a fish species

The wolf fish Hoplias malabaricus includes well differentiated sex systems (XY and X₁X₂Y in karyomorphs B and D, respectively), a nascent XY pair (karyomorph C) and not recognized sex chromosomes (karyomorph A). We performed the evolutionary analysis of these sex chromosomes, using two X chromosome-specific probes derived by microdissection from the XY and X₁X₂Y sex systems. A putative-sex pair in karyomorph A was identified, from which the differentiated XY system was evolved, as well as the clearly evolutionary relationship between the nascent XY system and the origin of the multiple X₁X₂Y chromosomes. The lack of recognizable signals on the sex chromosomes after the reciprocal cross-FISH experiments highlighted that they evolved independently from non-homologous autosomal pairs. It is noteworthy that these distinct pathways occur inside the same nominal species, thus exposing the high plasticity of sex chromosome evolution in lower vertebrates. Possible mechanisms underlying this sex determination liability are also discussed.     

Dioecious Silene at the X-road: the reasons Y

Among the variety of breeding systems developed by flowering plants, those based on heteromorphic sex chromosomes are the most intellectually challenging in evolutionary terms. This is because, among other things, they enable us to compare sex determination processes between plants and animals. White campion (Silene latifolia, also named Lychnis or Melandrium) is dioecious and, much like us, females are homogametic (XX) and males are heterogametic (XY). Sexual dimorphism in white campion is controlled by two independent developmental pathways operating from the Y chromosome at very early developmental stages and within distinct regions of the flower. In addition, all basic steps in the evolution from the bisexual to the dioecious condition with heteromorphic sex chromosomes are known and available to experimentation in the genus Silene. This group of species has been under scrutiny for more than a century. Such an ideal experimental system enables us to tackle, with novel methodological tools, several classical questions in biology. These include the question of how sexual dimorphism evolved and how dimorphic development is controlled, as well as questions of how sex chromosomes evolve in the absence of meiotic recombination or how male-female genetic conflicts are generated. At the turn of the century, the time is now ripe to have a closer look. Received: 21 September 1999 / Accepted: 11 October 1999     

HISTORICAL DIVERGENCE AND GENE FLOW: COALESCENT ANALYSES OF MITOCHONDRIAL,AUTOSOMAL AND SEX‐LINKED LOCI IN PASSERINA BUNTINGS

Quantifying the role of gene flow during the divergence of closely related species is crucial to understanding the process of speciation. We collected DNA sequence data from 20 loci (one mitochondrial, 13 autosomal, and six sex‐linked) for population samples of Lazuli Buntings (Passerina amoena) and Indigo Buntings (Passerina cyanea) (Aves: Cardinalidae) to test explicitly between a strict allopatric speciation model and a model in which divergence occurred despite postdivergence gene flow. Likelihood ratio tests of coalescent‐based population genetic parameter estimates indicated a strong signal of postdivergence gene flow and a strict allopatric speciation model was rejected. Analyses of partitioned datasets (mitochondrial, autosomal, and sex‐linked) suggest the overall gene flow patterns are driven primarily by autosomal gene flow, as there is no evidence of mitochondrial gene flow and we were unable to reject an allopatric speciation model for the sex‐linked data. This pattern is consistent with either a parapatric divergence model or repeated periods of allopatry with gene flow occurring via secondary contact. These results are consistent with the low fitness of female avian hybrids under Haldane's rule and demonstrate that sex‐linked loci likely are important in the initial generation of reproductive isolation, not just its maintenance.     

Chromosome banding in Amphibia

The mitotic and meiotic chromosomes of the marsupial frog Gastrotheca riobambae were analysed with various banding techniques. The karyotype of this species is distinguished by considerable amounts of constitutive heterochromatin and unusual, heteromorphic XY sex chromosomes. The Y chromosome is considerably larger than the X chromosome and almost completely heterochromatic. The analysis of the banding patterns obtained with GC- and AT-base-pair-specific fluorochromes shows that the constitutive heterochromatin in the Y chromosome consists of at least three different structural categories. The only nucleolus organizer region (NOR) of the karyotype is localized in the short arm of the X chromosome. This causes a sex-specific difference in the number of NOR: female animals have two NORs in diploid cells, male animals one. No cytological indications were found for the inactivation of one of the two X chromosomes in the female cells. In male meiosis, the heteromorphic sex chromosomes form a characteristic sex-bivalent by pairing their telomeres in an end-to-end arrangement. The significance of the XY/XX sex chromosomes of G. riobambae for the study of X-linked genes in Amphibia, the evolution of sex chromosomes and their specific DNA sequences, and the significance of the meiotic process of sex chromosomes are discussed.