The strength of reproductive isolation in two hybridizing food-deceptive orchid species

Reproductive isolation is of fundamental importance for maintaining species boundaries in sympatry. In orchids, the wide variety of pollination systems and highly diverse floral traits have traditionally suggested a prominent role for pollinator isolation, and thus for prezygotic isolation, as an effective barrier to gene flow among species. Here, we examined the nature of reproductive isolation between Anacamptis morio and Anacamptis papilionacea, two sister species of Mediterranean food-deceptive orchids, in two natural hybrid zones. Comparative analyses of the two hybrid zones that are located on soils with volcanic origin and have different and well-dated ages consistently revealed that all hybrid individuals were morphologically and genetically intermediate between the parental species, but had strongly reduced fitness. Molecular analyses based on nuclear ITS1 and (amplified fragment length polymorphism) AFLP markers clearly showed that all examined hybrids were F1 hybrids, and that no introgression occurred between parental species. The maternally inherited plastid DNA markers indicated that hybridization between A. morio and A. papilionacea was bidirectional, as confirmed by the molecular analysis of seed families. The genetic architecture of the two hybrid zones suggests that the two parental species easily and frequently hybridize in sympatry as a consequence of partial pollinator overlap but that strong postzygotic barriers reduce hybrid fitness and prevent gene introgression. These results corroborate that chromosomal divergence is instrumental for reproductive isolation between these food-deceptive orchids and suggest that hybridization is of limited importance for their diversification.     

Morphological variation of genetically confirmed Alouatta Pigra × A. palliata hybrids from a natural hybrid zone in Tabasco,Mexico

While hybridization has been reported for a large number of primate taxa, there is a general lack of data on hybrid morphology for wild individuals with known genetic ancestry. A confirmed hybrid zone for the closely related Neotropical primates Alouatta palliata and A. pigra has provided a unique opportunity to study primate hybrid morphological variation. Here we used molecular evidence based on mitochondrial, Y‐chromosome, and autosomal data to assess hybrid ancestry. We conducted univariate and multivariate statistical comparisons of morphometric data collected from individuals both outside and within the hybrid zone in Tabasco, Mexico. Our results show that of all the hybrids detected (N = 128), only 12% of them were approximately genetically intermediate, and none of them were first generation hybrids. Univariate pairwise comparisons among parental individuals, multigenerational backcrossed hybrids, and intermediate hybrids showed that overall, multigenerational backcrossed hybrids resemble the parental species with which they share most of their alleles. Conversely, intermediates were highly variable. Similarly, principal component analysis depicts an overlap between the parental species and their backcrosses when considering overall morphological differences. Finally, discriminant function analysis of the morphological variables was overall unreliable for classifying individuals into their assigned genotypic classes. Taken together, our results suggest that primate natural hybridization studies should incorporate molecular methods for determining ancestry, because morphology may not always be a reliable indicator of hybrid status. Hybrid zones could comprise a large number of multigenerational backcrossed hybrids that are indistinguishable from the parental species. The implications for studying hybridization in the primate fossil record are discussed. Am J Phys Anthropol, 2013. © 2012 Wiley Periodicals, Inc.     

Is the Prunella (Lamiaceae) hybrid zone structured by an environmental gradient? Evidence from a reciprocaltransplant experiment

Hybrid zones may be structured by environmentally independent selection against intrinsically unfit hybrids (tension zone models), or by environmentally dependent fitness differences among parental species and hybrids (ecological selection-gradient models). A 30-m slope in a mountain grassland harbors a hybrid zone of the clonal perennials, Prunella grandiflora and P. vulgaris (Lamiaceae), with P. grandiflora in the upper, P. vulgaris in the lower, and both parental species and P. grandiflora × P. vulgaris Hybrids in a narrow middle part. We found gradients for soil depth and water content, and vegetation height and biomass along the slope. A reciprocal transplant experiment yielded crossing reaction norms for vegetative reproduction. Parental species were locally adapted to their home sites, while the three taxa did not differ in vegetative reproduction in the Hybrid transplant site. Local adaptation for vegetative reproduction of P. grandiflora was mediated through higher survival and that of P. vulgaris through higher ramet number, indicating adaptation of their clonal growth strategies (phalanx vs. guerrilla) to the different habitats. Hybrid performance was intermediate between that of the parental species in all three sites, although Hybrids flowered more often than the parental species in the Hybrid site. Our results support ecological selection-gradient rather than tension zone models.     

Drought stress tolerance of Prosopis chilensis and Prosopis flexuosa species and their hybrids

Natural hybridization commonly produces individuals with intermediate morphological and genetic characteristics, but their response to environmental stress is still uncertain, with some studies showing that transgressive performance would be common. Prosopis chilensis and Prosopis flexuosa are the most important tree species from Arid Chaco, South-America. Both species occupy different ecological niches in terms of water availability. Genetic and morphological studies have demonstrated the existence of interspecific hybrids in contact areas between these species. Hybrids are characterized by clear intermediate morphological characteristics, which have taxonomical value, and genetic structure compared to both parental species. We studied mechanisms implicated in drought stress tolerance in seedlings of P. chilensis, P. flexuosa and their interspecific hybrids trying to elucidate if hybrids have a morpho-physiological, growth and survival intermediate response to drought compared to differential parental responses or if they out-perform both parental species when subjected to drought. Our results suggest that hybridization does not result in individuals with intermediate mechanisms related to drought resistance, but with a unique trait combination leading to high growth when water availability is high (similar to the most vulnerable parental species) and high survival under drought stress (similar to the more resistant parental species). Certain uncoupling between symplastic and apoplastic resistence to drought was observed in hybrids, as well as decreased physiological-wood anatomical plasticity compared to parental species. The long-term consequences in terms of adaptive response to drought of this particular trait combination of hybrids remain still unknown.     

POLLEN TRANSFER BY NATURAL HYBRIDS AND PARENTAL SPECIES IN AN IPOMOPSIS HYBRID ZONE

Models of hybrid zones differ in their assumptions about the relative fitnesses of hybrids and the parental species. These fitness relationships determine the form of selection across the hybrid zone and, along with gene flow, the evolutionary dynamics and eventual outcome of natural hybridization. We measured a component of fitness, export and receipt of pollen in single pollinator visits, for hybrids between the herbaceous plants Ipomopsis aggregata and I. tenuituba and for both parental species. In aviary experiments with captive hummingbirds, hybrid flowers outperformed flowers of both parental species by receiving more pollen on the stigma. Although hummingbirds were more effective at removing pollen from anthers of I. aggregata, hybrid flowers matched both parental species in the amount of pollen exported to stigmas of other flowers. These patterns of pollen transfer led to phenotypic stabilizing selection, during that stage of the life cycle, for a stigma position intermediate between that of the two species and to directional selection for exserted anthers. Pollen transfer between the species was high, with flowers of I. aggregata exporting pollen equally successfully to conspecific and I. tenuituba flowers. Although this study showed that natural hybrids enjoy the highest quality of pollinator visits, a previous study found that I. aggregata receives the highest quantity of pollinator visits. Thus, the relative fitness of hybrids changes over the life cycle. By combining the results of both studies, pollinator-mediated selection in this hybrid zone is predicted to be strong and directional, with hybrid fitness intermediate between that of the parental species.     

Genetic and morphological patterns show variation in frequency of hybrids between Ipomopsis (Polemoniaceae) zones of sympatry

Variation in rates of hybridization among zones of sympatry between a pair of species provides a useful window into the effect of local conditions on the evolution of reproductive isolation. We employed floral morphological traits and neutral genetic markers to quantify the frequency of individuals intermediate to the two parental species in two zones of sympatry between Ipomopsis aggregata and I. tenuituba, using clustering methods that make no a priori assumptions about population structure. The sites differed not only in the frequency of intermediate individuals, but also in climate, pollinator abundance and behavior and spatial structure of plant populations. Both floral traits, which are likely to be under natural selection and molecular markers, which are quasi-neutral, indicated more population structure at one site than the other, the pattern being more pronounced for floral morphology. One likely explanation for this difference between sites is that local ecological conditions, particularly pollinator choice of flowers, have promoted different rates of hybridization between these species. Hence, the evolution of reproductive isolation might depend in part on local conditions, and thus differ among populations of the same pair of species.     

Daphnia hybridization along ecological gradients in pelagic environments: the potential for the presence of hybrid zones in plankton

The relative homogeneity of pelagic environments has been regarded as the reason for the absence of hybrid zones for hybridizing planktonic Daphnia (Crustacea: Cladocera); occasional dominance of interspecific hybrids over parental species was explained by their temporal superiority in fluctuating environments. However, water bodies with spatially varying environmental conditions might facilitate the formation of hybrid zones in plankton. We studied the distribution of species and hybrids of the Daphnia longispina complex in 11 canyon-shaped reservoirs, localities characterized by horizontal environmental gradients (particularly of food supply and size-selective predation); we also analysed patterns of carapace size and fecundity among coexisting taxa. Spatial distribution of taxa agreed with their ecological characteristics; those showing different affinities along longitudinal reservoir profiles differed in size according to the presumed fish predation gradient. Only hybrids of Daphnia galeata with Daphnia cucullata and D. longispina (=hyalina) were recorded. The latter two species preferred opposite ends of gradients, such spatial segregation probably explaining the absence of their hybrids. Distributional patterns were relatively stable in two consecutive summers, apart from a substantial decline of D. galeata X cucullata in the second year. The observed pattern of a hybrid-dominated zone in intermediate conditions suggests that local Daphnia hybrid zones may indeed form within reservoirs.     

PATTERNS OF MATING IN WILD SUNFLOWER HYBRID ZONES

Theory predicts that homoploid hybrid speciation will be facilitated by selfing, yet most well-documented hybrid species are outcrossers. One possible explanation for this puzzle is that conditions in hybrid populations may favor selfing, even in otherwise outcrossing species. For example, in self-incompatible plants, mixtures of self and interspecific pollen often induce selfing. Here, we examine patterns of mating in three hybrid zones and four “pure” populations of Helianthus annuus and H. petiolaris, wild, self-incompatible sunflower species that are thought to have parented three homoploid hybrid species. Fourteen to 16 maternal families from each pure population and 44–46 maternal families from each hybrid zone were analyzed for seven polymorphic isozyme loci. Maximum-likelihood (ML) methods were used to estimate multilocus outcrossing rates (T_m) and hybridization frequencies for each maternal family, each phenotypic group within each hybrid zone (annuus-like, hybrid, and petiolaris-like), and each population. As predicted for self-incompatible species, all four parental populations have outcrossing rate ML estimates of 1.0. Within the hybrid zones, outcrossing rates were lowest in the H. annuus–like fraction of the population (0.73, 0.72, and 0.74 in the three hybrid zones, respectively), largely intermediate in the H. petiolaris–like group (0.94, 0.90, and 0.94), and highest in the hybrid group (0.97, 0.93, and 0.97). Although outcrossing rates are lower in hybrid zones than in pure populations, it is unlikely that the observed decrease facilitates hybrid speciation because outcrossing rates in the critical hybrid fraction of the population do not differ significantly from 1.0. Dividing the outcrossed pollen pool into intraspecific and interspecific components revealed that maternal plants are largely fertilized by conspecific pollen, confirming an important role for pollen competition as a reproductive barrier. Highly sterile hybrid plants do not appear to discriminate between parental species pollen, but hybrids with higher fertility tend to be fertilized by pollen from the parental group they resemble genetically. Thus, gametic selection leads to substantial assortative mating in these hybrid zones.     

The predictability of traits and ecological interactions on 17 different crosses of hybrid oaks

Herbivory on hybrid plants has the potential to affect patterns of plant evolution, such as limiting gene-flow through hybrids, and can also affect herbivore biodiversity. However, few studies have surveyed multiple hybrid species to identify phylogenetic patterns in the inheritance of plant traits that may drive herbivory. We surveyed 15 leaf traits and patterns of chewing, mining, and galling herbivory in a common garden of 17 artificially crossed hybrid oak species and each of their parental species over a 2-year period. Using a phylogeny of oaks, we tested whether hybrids that resulted from more divergent parents received more herbivory than those derived from closely related parents (as would be predicted by a build-up of incompatibilities in defensive systems over evolutionary time) and found only marginal evidence in support of this. We found that chewing damage to hybrids was weakly predicted by the relatedness of a parental species to the single native oak. The levels of chewing and mining herbivory on hybrids were typically intermediate to those of their parental species, though less than the parental mean for chewing damage in 2008. Most leaf traits of hybrids were also intermediate to those of their parental species. There was no clear pattern in terms of an association between 11 species of cynipid gall wasps and hybrids. The patterns of (1) intermediate levels of herbivory on hybrids and (2) no trend in herbivory on hybrids based on the phylogenetic relatedness of parental species suggest that herbivory may not play a general role in limiting hybrid fitness (and thus gene-flow through hybrids) in oaks.     

Hybrid songbirds employ intermediate routes in a migratory divide

Migratory divides are contact zones between populations that use different routes to navigate around unsuitable areas on seasonal migration. Hybrids in divides have been predicted to employ intermediate and potentially inferior routes. We provide the first direct test of this hypothesis, using light‐level geolocators to track birds breeding in a hybrid zone between Swainson's thrushes in western Canada. Compared to parental forms, hybrids exhibited increased variability in their migratory routes, with some using intermediate routes that crossed arid and mountainous regions, and some using the same routes as one parental group on fall migration and the other on spring migration. Hybrids also tended to use geographically intermediate wintering sites. Analysis of genetic variation across the hybrid zone suggests moderately strong selection against hybrids. These results indicate that seasonal migratory behaviour might be a source of selection against hybrids, supporting a possible role for migration in speciation.     

Evidence of gene flow between Sulawesi macaques

A pilot field study was conducted in Sulawesi (Indonesia) to assess the status of macaque populations on the island. Wild and captive animals were sampled, mainly in border areas between presumed different species. The five species investigated were Macaca maurus, M. tonkeana, M. hecki, M. nigrescens, and M. nigra, for which morphological and gene frequency data suggested the presence of hybridization zones. Some individuals within these zones showed intermediate or mosaic morphology between parental forms. These individuals also had intermediate gene frequencies for most of the polymorphic systems investigated. Karyotypes were identical in all species, and no cytogenetic barrier to hybridization existed between species. A review of the recent literature also provided evidence for hybridization between Sulawesi macaques. Clinical frequencies in both morphological and biomolecular traits perhaps can be best explained by the operation of gene flow between the various forms of macaques on the island. However, additional data are necessary before current classification schemes are revised. The unique opportunity and need of further study of Sulawesi macaques for a range of evolutionary questions is emphasized.     

Leaf morphological responses to variation in water availability for plants in the <Emphasis Type="Italic">Piriqueta caroliniana</Emphasis> complex

Distribution of plants and the expression of traits associated with environmental variation can be affected by both average conditions and the variance in conditions including extreme climatic events. We expect that these same factors should affect the distribution of plants in hybrid zones between ecologically distinct species where the hybrids should occupy ecotones or intermediate habitats. We evaluated water availability and leaf morphological differences among parental and hybrid populations of herbaceous perennial plants in the Piriqueta caroliniana complex along environmental gradients in Southeastern North America. We focus on two taxa in this group; the viridis morphotype, which occurs in southern Florida, and the caroliniana morphotype, which is distributed from northern Florida to southern Georgia. Advanced-generation hybrid derivatives of these morphotypes occupy a broad geographic region that extends across much of central Florida. Overall, we found that hybrid populations occurred in significantly drier locations, indicating that their habitat requirements are transgressive (i.e., exceeding parental values) rather than intermediate to the parental morphotypes. Water availability differed between the two sampling years, and plants displayed morphological changes in response to these changes in moisture. During the drier year, leaves were narrower and more hirsute, corroborating experimental results that these leaf traits are plastic, and confirming that plasticity occurs in natural habitats. Hybrids exhibited intermediate leaf traits (shape and size) across both years, and displayed transgressive (hair density) leaf traits during the drier year. The apparent canalization of the hybrids’ leaf morphological traits may contribute to their tolerance of variable environmental conditions and may partially explain why they have displaced the caroliniana morphotype in central Florida.     

Morphology, genetics, and ecology of pocket gophers (genus Geomys) in a narrow hybrid zone

A population of hybrid pocket gophers ( Geomys bursarius × G. lutescens ) exists 1–2 km west of Oakdale, Antelope County, Nebraska, U.S.A. The hybrids occur in soil that has characteristics intermediate between that occupied by G. lutescens (sand) and by G. bursarius (silt loam); the vegetation associations on the different soils are Sandhills Prairie and Tall-grass Prairie, respectively, with mixed prairie on the intermediate soils. Hybrids are identifiable on the basis of both qualitative and quantitative morphological characteristics, allozymes and karyology. Concordance between morphological, allozymic, karyotypic and ecological data sets is very high. Hybrids appear to reproduce normally and survive well; i.e. they suffer no obvious loss of fitness. Backcrossing to either parental type is apparently rare. The parental species each support obligate parasitic lice ( Geomydoecus : Trichodectidae) of different species; these species are not sister species. We suggest that hybrid zones resulting from primary and secondary contact may be distinguished by (1) concordance of clines in different character sets, (2) fossil and biogeographic data, and (3) parasite data. We conclude that this zone resulted from secondary contact, and that the zone is maintained either by selection against hybrids (less likely) or by hybrid superiority (more likely).     

Unexpected ancestry of Populus seedlings from a hybrid zone implies a large role for postzygotic selection in the maintenance of species

In the context of potential interspecific gene flow, the integrity of species will be maintained by reproductive barriers that reduce genetic exchange, including traits associated with prezygotic isolation or poor performance of hybrids. Hybrid zones can be used to study the importance of different reproductive barriers, particularly when both parental species and hybrids occur in close spatial proximity. We investigated the importance of barriers to gene flow that act early vs. late in the life cycle of European Populus by quantifying the prevalence of homospecific and hybrid matings within a mosaic hybrid zone. We obtained genotypic data for 11 976 loci from progeny and their maternal parents and constructed a Bayesian model to estimate individual admixture proportions and hybrid classes for sampled trees and for the unsampled pollen parent. Matings that included one or two hybrid parents were common, resulting in admixture proportions of progeny that spanned the whole range of potential ancestries between the two parental species. This result contrasts strongly with the distribution of admixture proportions in adult trees, where intermediate hybrids and each of the parental species are separated into three discrete ancestry clusters. The existence of the full range of hybrids in seedlings is consistent with weak reproductive isolation early in the life cycle of Populus. Instead, a considerable amount of selection must take place between the seedling stage and maturity to remove many hybrid seedlings. Our results highlight that high hybridization rates and appreciable hybrid fitness do not necessarily conflict with the maintenance of species integrity.     

Seed beetle survivorship,growth and egg size plasticity in a paloverde hybrid zone

1. Although numerous studies have examined the ecology of plant resistance to herbivores and the distribution of herbivores within plant hybrid zones, few have examined how plant hybridization influences herbivore growth, development, or life history. The experiment reported here examines variation in survivorship, development time, and final adult body size of Stator limbatus reared on seeds of parental and hybrid Cercidium floridum-×-C. microphyllum trees from a paloverde hybrid zone in eastern California. Because S. limbatus exhibits egg size plasticity in response to host species, the size of eggs that females lay on hybrid and parental plants is also examined. 2. The hypotheses (a) that seeds of hybrid trees are intermediate between those of the two parental species in their resistance to penetration by S. limbatus larvae; (b) that seeds of hybrid trees are intermediate in their suitability for the growth of larvae that successfully penetrate the seed-coat; (c) that female S. limbatus can distinguish between hybrid trees and pure-bred trees, as quantified by the size of eggs they lay on seeds of each taxon, and (d) that female S. limbatus can distinguish among individual hybrid trees, are tested. 3. On average, S. limbatus survivorship was lower, development time longer, and emergence body mass lower on seeds of C. floridum than on seeds of C. microphyllum. Seeds of hybrid trees were, on average (across trees), intermediate between seeds of the parental species in the resistance of their seed-coats to penetration by S. limbatus larvae and in their suitability for larval growth. Individual hybrid trees also varied in the resistance of their seeds to, and the suitability of their seeds for, S. limbatus larvae. 4. Female S. limbatus laid significantly larger eggs on seeds of C. floridum than on seeds of C. microphyllum, and, on average, intermediate size eggs on hybrid trees. The size of eggs laid by females also varied among hybrid trees, with females laying C. floridum-sized eggs on some trees, and C. microphyllum-sized eggs on other trees. These results suggest that females have at least some ability to distinguish among hybrid trees, but there was no evidence that females laid larger eggs on poorer quality hybrid trees.     

An ecomorphological framework for the coexistence of two cyprinid fish and their hybrids in a novel environment

Niche variation between hybrid taxa and their parental species has been deemed imperative to the persistence of hybrid populations in nature. However, the ecological factors promoting hybrid establishment remain poorly understood. Through the application of a multidisciplinary approach integrating genetics, morphometry, life‐history, and trophic ecology, we studied the hybrids of roach (Rutilus rutilus L.) and bream (Abramis brama L.), and their parental species inhabiting an Irish lake. The roach × bream hybrid exhibited a body shape intermediate of that of the parental species. Diet analyses depicted the hybrid as a generalist, feeding on all prey items consumed by either parental species. Stable isotope data confirm the trophic niche breadth of hybrids. A significant correlation between body shape and diet was detected, suggesting that the intermediate phenotype of hybrids might play a role in their feeding abilities, resulting in the utilization of a broader trophic spectrum than the parental species. Growth and age class structure analyses also yielded a scenario that is consistent with the ecological success of hybrids. Genetic analyses suggest that the majority of hybrids result from first‐generation crosses between the parental species; however, a potentially significant proportion of back‐crosses with bream were also detected. The recent introduction of roach and bream into Irish waters, as well as the climatic and ecological features of the colonized habitats, can explain the remarkable success of the roach × bream hybrid in Ireland. The adaptive significance of hybridization and its demographic consequences for the parental species are discussed. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 768–783.     

Recombinant hybrids retain heterozygosity at many loci: new insights into the genomics of reproductive isolation in Populus

The maintenance of species barriers in the face of gene flow is often thought to result from strong selection against intermediate genotypes, thereby preserving genetic differentiation. Most speciation genomic studies thus aim to identify exceptionally divergent loci between populations, but divergence will be affected by many processes other than reproductive isolation (RI) and speciation. Through genomic studies of recombinant hybrids sampled in the wild, genetic variation associated with RI can be observed in situ, because selection against incompatible genotypes will leave detectable patterns of variation in the hybrid genomes. To better understand the mechanisms directly involved in RI, we investigated three natural ‘replicate’ hybrid zones between two divergent Populus species via locus‐specific patterns of ancestry across recombinant hybrid genomes. As expected, genomic patterns in hybrids and their parental species were consistent with the presence of underdominant selection at several genomic regions. Surprisingly, many loci displayed greatly increased between‐species heterozygosity in recombinant hybrids despite striking genetic differentiation between the parental genomes, the opposite of what would be expected with selection against intermediate genotypes. Only a limited, reproducible set of genotypic combinations was present in hybrid genomes across localities. In the absence of clearly delimited ‘hybrid habitats’, our results suggest that complex epistatic interactions within genomes play an important role in advanced stages of RI between these ecologically divergent forest trees. This calls for more genomic studies that test for unusual patterns of genomic ancestry in hybridizing species.     

Behavioral sterility of hybrid males in acoustically communicating grasshoppers (Acrididae,Gomphocerinae)

The effectiveness of hybridization barriers determines whether two species remain reproductively isolated when their populations come into contact. We investigated acoustic mating signals and associated leg movements responsible for song creation of hybrids between the grasshopper species Chorthippus biguttulus and C. brunneus to study whether and how songs of male hybrids contribute to reproductive isolation between these sympatrically occurring species. Songs of F1, F2, and backcross hybrids were intermediate between those of both parental species in terms phrase number and duration. In contrast, species-specific syllable structure within phrases was largely lost in hybrids and was produced, if at all, in an irregular and imperfect manner. These divergences in inheritance of different song parameters are likely the result of incompatibility of neuronal networks that control stridulatory leg movements in hybrids. It is highly probable that songs of hybrid males are unattractive to females of either parental species because they are intermediate in terms of phrase duration and lack a clear syllable structure. Males of various hybrid types (F1, F2, and backcrosses) are behaviorally sterile because their songs fail to attract mates.     

Reticulate evolution on a mosaic of soils: diversification of the New Caledonian endemic genus Codia (Cunoniaceae)

We reconstructed the evolutionary history of Codia , a plant genus endemic to the New Caledonia biodiversity hotspot in the southwest Pacific, using three single-copy nuclear genes. It seems likely that more than half of Codia species have a hybrid origin, but in the absence of cytological information, it is not known whether polyploids occur. Adaptation to ultramafic soils is possibly a plesiomorphic character for the entire genus. We found that species of hybrid origin can have some morphological characters absent in putative parental species, that is, they exhibit transgressive phenotypes. There is evidence of considerable range alteration post-origin in several species because some likely parental species of hybrid taxa no longer co-occur and are confined to putative rainforest refugia; in some cases, hybrid species do not now co-occur with either of their parental species. These results have implications for the design of conservation strategies, for example, prioritization of parental species for ex-situ conservation and preservation of the contact zones between soil types where hybridization is more likely to occur (i.e. conserving the possibility for the process to continue rather than trying to conserve taxa).     

Morphological distinctiveness of Ligularia tongolensis and L. cymbulifera is maintained between habitats despite bidirectional and asymmetrical introgression in multiple hybrid zones

Natural hybridization is a crucial evolutionary process and a long-standing topic of study in evolutionary biology. Hybrid zones, where two congeneric species interact, can provide insight into the process of natural hybridization, especially with respect to how taxon diversity is maintained. In this study, we used double digest restriction-site associated DNA sequencing technology (ddRAD-seq) to examine genetic structure and estimate introgression in four hybrid zones of Ligularia tongolensis and Ligularia cymbulifera. Our analysis demonstrated that parental species were highly differentiated, whereas pairwise F_ST between parents and their hybrids was low, indicating that sympatric sites can form hybrid zones. As most F₁ hybrid individuals were observed within these zones, our finding also implied the presence of substantial barriers to interbreeding. Furthermore, some individuals that possessed the typical morphology of the parental species belonged to the F₁ generation. Genomic clines analysis revealed that a large fraction of single nucleotide polymorphisms (SNPs) deviated from a model of neutral introgression in the four hybrid zones, and most SNPs exhibited selection favoring the L. cymbulifera genotype. Bidirectional but asymmetric introgression was revealed as evident in the four hybrid zones. Habitat differences between the four hybrid zones may affect isolation barriers between both species. Taken together, these findings suggest that where incomplete reproductive barriers allow natural hybridization, the introgression between species generates rich genetic recombination that contributes to the fast adaptation and diversification of the widespread Ligularia in the Hengduan Mountains Region (HMR).