Are linear‐leaved Potamogeton hybrids really so rare? Molecular evidence for multiple hybridizations between P. acutifolius and P. compressus in central Europe

Due to insufficient morphological differentiation, hybridization among linear‐leaved species of Potamogeton is rarely observed and recognized. Here, we applied molecular tools (sequencing of nrITS and rpl32‐trnL cpDNA intergenic spacer) to study several natural, central European populations of morphologically intermediate forms between two closely related species, P. acutifolius and P. compressus, to examine their taxonomical status and test whether they represented the variation range of the species or are results of interspecific hybridization. Both DNA regions provided distinguishing characters between putative parental species. The ITS sequences from all the morphologically intermediate specimens displayed an additive pattern combining the variation of both parental species, providing evidence for the hybrid origin of all these plants. This case study suggests that hybrids of linear‐leaved Potamogeton species are more common than usually believed, but have been difficult to identify without molecular tools.     

Using DNA‐based techniques to identify hybrids among linear‐leaved Potamogeton plants collected in China

Abstract It is well known that interspecific hybrids occur in the genus Potamogeton. The linear‐leaved Potamogeton species commonly have highly variable morphological characteristics. Their hybrids often show similar vegetative characters to their parental species and their identification based solely on morphology is not always conclusive. In order to clarify whether there are any hybrids from the linear‐leaved Potamogeton plants collected in China, we used internal transcribed spacers (ITS) of nuclear ribosomal DNA and chloroplast rbcL gene sequences to identify the hybrids. Using ITS sequence additivity, we identified four hybrids, namely P. orientalis (P. pusillus×P. oxyphyllus), P. pusillus×P. berchtoldii, P. foliosus×P. octandrus, and P. cristatus×P. octandrus. The latter three hybrids should be considered as new hybrids in Potamogeton. The maternal parents of the four hybrids were confirmed using chloroplast rbcL gene sequences.     

Persistence of transgenes in wild rice populations depends on the interaction between genetic background of recipients and environmental conditions

The persistence of transgenes in wild populations may cause unintended ecological consequences, and the possibility of transgenes' persistence and introgression is dependent on fitness performance of transgenic crop–wild hybrids. To investigate the effects of transgene and genotype × environment on the fitness of crop–wild rice hybrids, a total of 11 cross‐combination progenies between insect‐resistant transgene (CpTI and Bt/CpTI) rice lines and wild rice (Oryza rufipogon) were evaluated at different sites with contrasting insect treatments. The results showed that fitness performance varied between transgenic hybrids having different wild parents and under different environmental conditions, indicating that fitness effects of transgenes on hybrid progenies depend heavily on the genetic background of recipient plants and growing environment. Significant fitness advantages conferred by transgenes were found only in some hybrids under high insect pressure condition, demonstrating that the level of target insects in the field environment influences the persistence and spread of insect‐resistant transgenes in wild rice populations. These findings suggest that evolutionary fate of escaped transgenes is different in wild populations with diverse genetic backgrounds under various environmental conditions.     

Diagnostic SNPs reveal widespread introgressive hybridization between introduced bighead and silver carp in the Mississippi River Basin

Hybridization among conspecifics in native and introduced habitats has important implications for biological invasions in new ecosystems. Bighead (Hypophthalmichthys nobilis) and silver carp (H. molitrix) are genetically isolated and occur in sympatry within their native range. Following their introduction to North America, however, introgressant hybrids have been reported throughout their expanded range within the Mississippi River Basin (MRB). The extent of introgression, both spatially and generationally, is largely unknown. Therefore, we examined mixed‐species populations from across the MRB to characterize the extent of interspecific gene flow. We assayed 2798 individuals from nine locations with a suite of species‐diagnostic SNPs (57 nuclear and one mitochondrial). Forty‐four per cent (n = 1244) of individuals displayed hybrid genotypes. Moreover, the composition of hybrid genotypes varied among locations and represented complex hybrid swarms with multiple generations of gene flow. Introgressive hybrids were identified from all locations, were bidirectional and followed a bimodal distribution consisting primarily of parental or parental‐like genotypes and phenotypes. All described hybrid categories were present among individuals from 1999 to 2008, with parents and later‐generation backcrosses representing the largest proportion of individuals among years. Our mitochondrial SNP (COII), tested on a subset of 730 individuals, revealed a silver carp maternal bias in 13 of 21 (62%) F₁ hybrids, in all silver carp backcrosses, and maintained throughout many of the bighead carp backcrosses. The application of this suite of diagnostic markers and the spatial coverage permits a deeper examination of the complexity in hybrid swarms between two invasive, introduced species.     

Influence of niche similarity on hybridization between Myriophyllum sibiricum and M. spicatum

The impact of ecological factors on natural hybridization is of widespread interest. Here, we asked whether climate niche influences hybridization between the two closely related plant species Myriophyllum sibiricum and M. spicatum. Eight microsatellite loci and two chloroplast fragments were used to investigate the occurrence of hybridization between these two species in two co‐occurring regions: north‐east China (NEC) and the Qinghai‐Tibetan Plateau (QTP). The climate niches of the species were quantified by principal component analysis with bioclimatic data, and niche comparisons were performed between the two species in each region. Reciprocal hybridization was observed, and M. sibiricum was favoured as the maternal species. Furthermore, hybrids were rare in NEC but common in the QTP. Accordingly, in NEC, the two species were climatically distinct, and hybrids only occurred in the narrow geographical or ecological transition zone, whereas in the QTP, obvious niche overlaps were found for the two species, and hybrids occurred in multiple contact zones. This association between hybridization pattern and climate niche similarity suggests that the level of hybridization was promoted by niche overlap. Compared with the parental species, similar climate niches were found for the hybrid populations in the QTP, indicating that other environmental factors rather than climate were important for hybrid persistence. Our findings highlight the significance of climate niche with respect to hybridization patterns in plants.     

Genetic Evidence for Natural Hybridization between Species of Dioecious Ficus on Island Populations1

Natural hybrids between Ficus septica and two closely related dioecious species, F. fistulosa and F. hispida, were confirmed using amplified fragment length polymorphisms (AFLP) and chloroplast DNA markers. Ficus species have a highly species‐specific pollination mutualism with agaonid wasps. Therefore, the identification of cases in which breakdown in this sophisticated system occurs and the circumstances under which it happens is of interest. Various studies have confirmed that Ficus species are able to hybridize and that pollinator‐specificity breakdown can occur under certain conditions. This study is the first example in which hybrid identity and the presence of hybrids in the natural distribution of parental species for Ficus have been confirmed with molecular markers. Hybrid individuals were identified on three island locations in the Sunda Strait region of Indonesia. These findings support Janzen's (1979) hypothesis that breakdown in pollinator specificity is more likely to occur on islands. We hypothesized that hybrid events could occur when the population size of pollinator wasps was small or had been small in one of the parental species. Later generation hybrids were identified, indicating that backcrossing and introgression did occur to some extent and that therefore, hybrids could be fertile. The small number of hybrids found indicated that there was little effect of hybridization on parental species integrity over the study area. Although hybrid individuals were not common, their presence at multiple sites indicated that the hybridization events reported here were not isolated incidences. Chloroplast DNA haplotypes of hybrids were not derived solely from one species, suggesting that the seed donor was not of the same parental species in all hybridization events.     

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.     

ECOLOGICAL SEGREGATION IN A SMALL MAMMAL HYBRID ZONE: HABITAT‐SPECIFIC MATING OPPORTUNITIES AND SELECTION AGAINST HYBRIDS RESTRICT GENE FLOW ON A FINE SPATIAL SCALE

The degree to which closely related species interbreed is determined by a complex interaction of ecological, behavioral, and genetic factors. We examine the degree of interbreeding between two woodrat species, Neotoma bryanti and N. lepida, at a sharp ecological transition. We identify the ecological association of each genotypic class, assess the opportunity for mating between these groups, and test whether they have similar patterns of year‐to‐year persistence on our study site. We find that 13% of individuals have a hybrid signature but that the two parental populations and backcrosses are highly segregated by habitat type and use. Also, we find that adult hybrids are comparable to parental types in terms of year‐to‐year persistence on our site but that, among juveniles, significantly fewer hybrids reach adulthood on site compared to their purebred counterparts. Our analyses show that this hybrid zone is maintained by occasional nonassortative mating coupled with hybrid fertility, but that these factors are balanced by lower apparent survival of juvenile hybrids and habitat‐based preference or selection that limits heterospecific mating while promoting backcrossing to habitat‐specific genotypes. This system presents a novel example of the role that sharp resource gradients play in reproductive isolation and the potential for genetic introgression.     

A test of hybrid growth disadvantage in wild, free-ranging species pairs of threespine stickleback (Gasterosteus aculeatus) and its implications for ecological speciation

Ecological speciation is the evolution of reproductive isolation as a direct or indirect consequence of divergent natural selection. Reduced performance of hybrids in nature is thought to be an important process by which natural selection can favor the evolution of assortative mating and drive speciation. Benthic and limnetic sympatric species of threespine stickleback (Gasterosteus aculeatus) are adapted to alternative trophic niches (bottom browsing vs. open water planktivory, respectively) and reduced feeding performance of hybrids is thought to have contributed to the evolution of reproductive isolation. We tested this “hybrid‐disadvantage hypothesis” by inferring growth rates from otoliths sampled from wild, free‐ranging benthic, limnetic, and hybrid sticklebacks in two lakes. There were significant differences in growth rate between lakes, life‐history stages, and among years (maximum P = 0.02), as well as interactions between most factors, but not between hybrid and parental species sticklebacks in most comparisons. Our results provide little evidence of a growth disadvantage in hybrid sticklebacks when free‐ranging in nature. Although trophic ecology per se may contribute less to ecological speciation than envisioned, it may act in concert with other aspects of stickleback biology, such as interactions with parasites, predators, competitors, and/or sexual selection, to present strong multifarious selection against hybrids.     

Expression of anatomical leaf traits in homoploid hybrids between deciduous and evergreen species of Vaccinium

We investigated the anatomical expression of leaf traits in hybrids between evergreen Vaccinium vitis‐idaea and deciduous V. myrtillus. We compared parents from four populations with their respective F1 hybrids and tested whether (i) transgression can be the source of novel anatomical traits in hybrids; (ii) expression of transgressive traits is more probable for traits with similar values in parents and intermediate for more distinct values, as predicted by theory; and (iii) independent origin of hybrids leads to identical trait expression profiles among populations. We found that anatomical leaf traits can be divided into four categories based on their similarity to parents: intermediate, parental‐like, transgressive and non‐significant. Contrary to the common view, parental‐like trait values were equally important in shaping the hybrid profile, as were intermediate traits. Transgression was revealed in 17/144 cases and concerned mainly cell and tissue sizes. As predicted by theory, we observed transgressive segregation more often when there was little phenotypic divergence, but intermediate values when parental traits were differentiated. It is likely that cell and tissue sizes are phylogenetically more conserved due to stabilising selection, whereas traits such as leaf thickness and volume fraction of the intercellular spaces, showing a consistent intermediate pattern across populations, are more susceptible to directional selection. Hybrid populations showed little similarity in expression profile, with only three traits identically expressed across all populations. Thus local adaptation of parental species and specific genetic background may be of importance.     

Genetic markers for the identification of hybrids among catfish species of the family Pimelodidae

Fish hybrids provide genetically manipulated products of excellent value for the commercial aquaculture industry. However, if handled or marketed incorrectly, they can cause great financial loss to producers as well as threaten the native species. Herein, molecular markers are established to identify hybrid lineages of pimelodids and characterize them in relation to their parental species, Pseudoplatystoma corruscans, Pseudoplatystoma reticulatum, Phractocephalus hemioliopterus and Leiarius marmoratus. The results show that the mitochondrial genes are useful for identification of the cross‐direction through the characterization of the maternal lineage. The nuclear genes allow identification of the interspecific hybrids. Use of genetic markers can avoid misidentification of hybrids that occur in simple morphological analysis. Thus, the present results allow the routine monitoring of pimelodid hybrids for their correct management and trade in aquaculture.     

Post‐zygotic isolation in cactophilic Drosophila: larval viability and adult life‐history traits of D. mojavensis/D. arizonae hybrids

Drosophila mojavensis and Drosophila arizonae are cactophilic flies that have been used extensively in speciation studies. Incomplete premating isolation, evidence of reinforcement, and a lack of recent introgression between these species point to a potentially important role for post‐zygotic isolating barriers in this system. Other than hybrid male sterility, however, post‐zygotic isolation between D. mojavensis and D. arizonae has received little attention. In this study, we examined viability and life‐history traits of D. mojavensis/D. arizonae F₁ hybrids from sympatric crosses. Specifically, we reared hybrids and purebreds on the natural host cacti of each parental species and compared viability, development time, thorax length, and desiccation resistance between hybrids and purebreds. Interestingly, hybrid females from both crosses performed similarly or even better than purebred females. In contrast, hybrid sons of D. arizonae mothers, in addition to being sterile, had shorter average thorax length than males of both parental species, and hybrid males from both crosses had substantially lower desiccation resistance than D. mojavensis males. The probable cost to hybridization for D. mojavensis females resulting from reduced desiccation resistance of hybrid sons may have been an important selective factor in the history of reinforcement for crosses involving these females.     

Morphological and genetic evidence of contemporary intersectional hybridisation in Mediterranean Helichrysum (Asteraceae,Gnaphalieae)

Hybridisation is considered an important evolutionary phenomenon in Gnaphalieae, but contemporary hybridisation has been little explored within the tribe. Here, hybridisation between Helichrysum orientale and Helichrysum stoechas is studied at two different localities in the islands of Crete and Rhodes (Greece). Using three different types of molecular data (AFLP, nrDNA ITS sequences and cpDNA ndhF sequences) and morphological data, the aim is to provide simultaneous and direct comparisons between molecular and morphological variation among the parental species and the studied hybrid populations. AFLP profiles, ITS sequences and morphological data support the existence of hybrids at the two localities studied, shown as morphological and genetic intermediates between the parental species. Chloroplast DNA sequences show that both parental species can act either as pollen donor or as maternal parent. Fertility of hybrids is demonstrated by the viability of seeds produced by hybrids from both localities, and the detection of a backcross specimen to H. orientale. Although there is general congruence of morphological and molecular data, the analysis of morphology and ITS sequences can fail to detect backcross hybrids.     

Variation of plastic responses to oxygen availability within the hybridogenetic Rana esculenta complex

The hypothesis of superiority of hybrid phenotype to explain the maintenance of hybridogenesis (exclusion of one parental genome in the germline before meiosis) was investigated in the Rana esculenta complex. Survival and growth of progenies from each parental and hybrid lineages were compared across contrasted conditions of oxygen availability (this factor is suspected to influence habitat utilization of each form of the complex). Whereas growth and development were strongly affected by constant hypoxic conditions in the R. ridibunda lineage, they were not altered in the R. lessonae lineage. Because the performance of the hybrid (R. esculenta) lineage proved to be intermediate between the parental species, no heterosis effect was detected. These results supported the intermediate niche hypothesis as an alternative to the heterotic hybrid superiority hypothesis. The ecological and evolutionary implications are then discussed.     

Testing hybridization hypotheses and evaluating the evolutionary potential of hybrids in mangrove plant species

Natural hybridization is of marked importance from global to local biological diversity. In mangroves, species ranges overlap extensively with one another and species share a long overlap of flowering time. Although hybridization has been suggested, patterns of hybridization and the evolutionary potential of hybrids are not yet fully understood. This study provides molecular evidence for the parental origins and status of hybrids in the dominant mangrove genus Rhizophora based on comparisons of chloroplast and nuclear phylogenies and estimations of genetic relatedness and structure from inter‐simple sequence repeat (ISSR) markers. Phylogenetic analyses indicate that almost all species can act as maternal parents to hybrids and that hybridization can be bidirectional. Bayesian analyses indicate that hybrids are simple F₁s, and no trace of backcrossing was detected within populations. Hybridization, for the most part, occurs almost only locally and dispersal of hybrid individuals is limited beyond the hybrid sites.     

Different mechanisms for Arabidopsis thaliana hybrid necrosis cases inferred from temperature responses

Temperature is a major determinant of plant growth, development and success. Understanding how plants respond to temperature is particularly relevant in a warming climate. Plant immune responses are often suppressed above species‐specific critical temperatures. This is also true for intraspecific hybrids of Arabidopsis thaliana that express hybrid necrosis due to inappropriate activation of the immune system caused by epistatic interactions between alleles from different genomes. The relationship between temperature and defence is unclear, largely due to a lack of studies that assess immune activation over a wide range of temperatures. To test whether the temperature‐based suppression of ectopic immune activation in hybrids exhibits a linear or non‐linear relationship, we characterised the molecular and morphological phenotypes of two different necrotic A. thaliana hybrids over a range of ecologically relevant temperatures. We found both linear and non‐linear responses for expression of immunity markers and for morphological defects depending on the underlying genetic cause. This suggests that the influence of temperature on the trade‐off between immunity and growth depends on the specific defence components involved.     

The role of parental and hybrid species in multiple introgression events: evidence of homoploid hybrid speciation in Centaurea (Cardueae,Asteraceae)

Molecular approaches have greatly increased the number of confirmed homoploid hybrids, which suggests that the frequency of this phenomenon was underestimated in the past because it was much more difficult to detect than allopolyploidy. Centaurea is a suitable model group for studying homoploid speciation, as hybridization events have been commonly reported for this genus. Based on this, here we study Centaurea × forsythiana, a naturally occurring homoploid hybrid between two Sardinian endemics, C. horrida and C. filiformis, using a molecular approach involving nuclear and plastid markers, to understand the underlying population dynamics between homoploid hybrids and their parents. Our results confirm that C. × forsythiana is a hybrid between the above‐mentioned species and define the roles of the parents. Plastid markers point towards C. horrida as the maternal progenitor, and nuclear markers reveal that the other parental species, C. filiformis, is itself an old, stabilized homoploid hybrid related to the C. paniculata complex from the Italian mainland. Homoploid hybrid speciation is discussed and C. × forsythiana and C. filiformis are compared with other similar examples. The study confirms the importance of introgression between parental species mediated by hybrids and its potential implications in conservation. Furthermore, it shows how hybridization studies become even more complex when the parents are themselves of probable hybrid origin. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 453–467.     

Widespread hybridization within mound‐building wood ants in Southern Finland results in cytonuclear mismatches and potential for sex‐specific hybrid breakdown

Hybridization and gene flow between diverging lineages are increasingly recognized as common evolutionary processes, and their consequences can vary from hybrid breakdown to adaptive introgression. We have previously found a population of wood ant hybrids between Formica aquilonia and F. polyctena that shows antagonistic effects of hybridization: females with introgressed alleles show hybrid vigour, whereas males with the same alleles show hybrid breakdown. Here, we investigate whether hybridization is a general phenomenon in this species pair and analyse 647 worker samples from 16 localities in Finland using microsatellite markers and a 1200‐bp mitochondrial sequence. Our results show that 27 sampled nests contained parental‐like gene pools (six putative F. polyctena and 21 putative F. aquilonia) and all remaining nests (69), from nine localities, contained hybrids of varying degrees. Patterns of genetic variation suggest these hybrids arise from several hybridization events or, instead, have backcrossed to the parental gene pools to varying extents. In contrast to expectations, the mitochondrial haplotypes of the parental species were not randomly distributed among the hybrids. Instead, nests that were closer to parental‐like F. aquilonia for nuclear markers preferentially had F. polyctena's mitochondria and vice versa. This systematic pattern suggests there may be underlying selection favouring cytonuclear mismatch and hybridization. We also found a new hybrid locality with strong genetic differences between the sexes similar to those predicted under antagonistic selection on male and female hybrids. Further studies are needed to determine the selective forces that act on male and female genomes in these newly discovered hybrids.     

WIDESPREAD HOST‐DEPENDENT HYBRID UNFITNESS IN THE PEA APHID SPECIES COMPLEX

Linking adaptive divergence to hybrid unfitness is necessary to understand the ecological factors contributing to reproductive isolation and speciation. To date, this link has been demonstrated in few model systems, most of which encompass ecotypes that occupy relatively early stages in the speciation process. Here we extend these studies by assessing how host‐plant adaptation conditions hybrid fitness in the pea aphid, Acyrthosiphon pisum. We made crosses between and within five pea aphid biotypes adapted to different host plants and representing various stages of divergence within the complex. Performance of F1 hybrids and nonhybrids was assessed on a “universal” host that is favorable to all pea aphid biotypes in laboratory conditions. Although hybrids performed equally well as nonhybrids on the universal host, their performance was much lower than nonhybrids on the natural hosts of their parental populations. Hence, hybrids, rather than being intrinsically deficient, are maladapted to their parents’ hosts. Interestingly, the impact of this maladaptation was stronger in certain hybrids from crosses involving the most divergent biotype, suggesting that host‐dependent postzygotic isolation has continued to evolve late in divergence. Even though host‐independent deficiencies are not excluded, hybrid maladaptation to parental hosts supports the hypothesis of ecological speciation in this complex.     

The production of hybrids with high ecological amplitude between exotic Spartina densiflora and native S. maritima in the Iberian Peninsula

Aim Hybridization is a common and potent mechanism of plant evolution that has the potential to be evolutionary significant in its own right, and hybrids are common between invasive and native congeneric species. Our aims were to document the existence and nature of new Spartina hybrids arising between introduced Spartina densiflora and native S. maritima in tidal marshes of the Iberian Peninsula, to examine the actual and potential range of hybrids in the intertidal zone, and to analyse the seed set of hybrids. Location South‐West Iberian Peninsula. Methods Hybrids were characterized using chloroplast and nuclear DNA, and ploidy assessments. The ecological tolerance of the hybrids was studied using vegetation surveys and transplant experiments. Results We found that both parental species have been seed parents to hybrids and that all hybrids had an additive pattern of species‐specific nuclear markers consistent with F₁ hybrids. Hybrid chromosome numbers varied between 2n = ca. 65 and 2n = ca. 94–97, while S. maritima had 2n = ca. 60 and S. densiflora had 2n = ca. 70. Hybrids grew in three discrete locations along the intertidal zone but were capable of growing throughout the ranges of both parental species in transplanted gardens, and in most cases, grew better than the parental species. While the potential exists for the origination of another invasive Spartina hybrid species, thus far hybrid plants are not fertile, limiting their ability to invade and spread. Main conclusions We recommend the eradication of all of the currently quite limited hybrid cordgrass and to fight the S. densiflora invasion in the Iberian Peninsula to prevent the origination of a new invasive allopolyploid Spartina species.