Does intraspecific hybridization contribute to the evolution of invasiveness?: an experimental test

One of the major objectives of research on invasive species is to determine the relative importance of different evolutionary and ecological forces in the invasion process. It was recently suggested that post-introduction intraspecific hybridization between previously isolated genotypes could produce novel and/or heterotic progeny that might express enhanced invasiveness. We tested this hypothesis with Silene latifolia, a European native that has successfully invaded North America and has previously been shown to have undergone genetic change since its introduction. In a common garden experiment we compared the performance of plants derived from within and between population crosses from eight European and 18 North American populations. Results showed that there was no significant effect of crossing distance on progeny phenotype. Furthermore, progeny from within or between population crosses did not differ in size, reproductive output or survival. Collectively, these results suggest that the invasive phenotype of S. latifolia is likely the result of natural selection and/or genetic drift rather than intraspecific hybridization.     

Intra- and interspecific hybridization in invasive Siberian elm

Hybridization creates unique allele combinations which can facilitate the evolution of invasiveness. Frequent interspecific hybridization between the Siberian elm, Ulmus pumila, and native elm species has been detected in the Midwestern United States, Italy and Spain. However, Ulmus pumila also occurs in the western United States and Argentina, regions where no native elm species capable of hybridizing with it occurs. We examined whether inter- or intraspecific hybridization could be detected in these regions. Nuclear markers and the program STRUCTURE helped detect interspecific hybridization and determine the population genetic structure in both the native and the two non-native ranges. Chloroplast markers identified sources of introduction into these two non-native ranges. No significant interspecific hybridization was detected between U. pumila and U. rubra in the western United States or between U. pumila and U. minor in Argentina and vice versa. However, the genetic findings supported the presence of intraspecific hybridization and high levels of genetic diversity in both non-native ranges. The evidence presented for intraspecific hybridization in the current study, combined with reports of interspecific hybridization from previous studies, identifies elm as a genus where both inter- and intraspecific hybridization may occur and help maintain high levels of genetic diversity potentially associated with invasiveness.     

Reproductive success of cultivated Pyrus calleryana (Rosaceae) and establishment ability of invasive, hybrid progeny

?Premise of the study: Pyrus calleryana Decne., an ornamental tree species introduced from China, is a relatively new invasive that has only recently begun to spread across the United States after intraspecific hybridization between cultivars. The function of such hybridization in the evolution of invasiveness is still relatively understudied, especially with respect to the initial establishment and persistence of invasive genotypes. Multiple introductions of genetically divergent populations or cultivars may benefit from new genetic combinations created during hybridization events and/or release from Allee effects in founder populations. ?Methods: We quantified the outcome of intraspecific hybridization between cultivars of P. calleryana in a common garden. Measures of the reproductive success and establishment ability of their early- and advanced-generation hybrid offspring were collected to assess the likelihood of particular cultivar genotypes to establish in invasive populations. These traits also were compared between cultivated and invasive parents to identify any generational differences in invasive potential. ?Key results: Differences were detected in measures of reproductive ability, but no group emerged as consistently more fecund. Advanced-generation hybrids also had significantly less biomass, indicating a reduction in hybrid performance relative to that of the cultivated progeny. ?Conclusion: Ultimately, this study indicates that increased spread of P. calleryana has been initiated by introduction of multiple cultivar types and subsequent widespread planting and is not the result of an inherent fitness advantage of hybrid progeny.     

Hybridization and the colonization of novel habitats by annual sunflowers

Although invasive plant species often have a hybrid ancestry, unambiguous evidence that hybridization has stimulated the evolution of invasive behaviors has been difficult to come by. Here, we briefly review how hybridization might contribute to the colonization of novel habitats, range expansions, and invasiveness and then describe work on hybrid sunflowers that forges a direct link between hybridization and ecological divergence. We first discuss the invasion of Texas by the common sunflower and show that the introgression of chromosomal segments from a locally adapted species may have facilitated range expansion. We then present evidence that the colonization of sand dune, desert floor, and salt marsh habitats by three hybrid sunflower species was made possible by selection on extreme or “transgressive” phenotypes generated by hybridization. This body of work corroborates earlier claims regarding the role of hybridization in adaptive evolution and provides an experimental and conceptual framework for ongoing studies in this area.     

Molecular analysis of <Emphasis Type="Italic">Ixodes granulatus</Emphasis>, a possible vector tick for <Emphasis Type="Italic">Borrelia burgdorferi</Emphasis> sensu lato in Taiwan

The genetic identity of Ixodes granulatus ticks was determined for the first time in Taiwan. The phylogenetic relationships were analyzed by comparing the sequences of mitochondrial 16S ribosomal DNA gene obtained from 19 strains of ticks representing seven species of Ixodes and two outgroup species (Rhipicephalus sanguineus and Haemaphysalis inermis). Four major clades could be easily distinguished by neighbour-joining analysis and were congruent by maximum-parsimony method. All these I. granulatus ticks of Taiwan were genetically affiliated to a monophyletic group with highly homogeneous sequences (92.2–99.3% similarity), and can be discriminated from other Ixodes species and other genera of ticks with a sequence divergence ranging from 11.7 to 30.8%. Moreover, intraspecific analysis revealed that two distinct lineages are evident between the same species of I. granulatus ticks collected from Taiwan and Malaysia. Our results demonstrate that all these I. granulatus ticks of Taiwan represent a unique lineage distinct from the common vector ticks (I. ricinus complex) for Borrelia burgdorferi spirochetes.     

The leafy liverwortPorella baueri (Porellaceae) is an allopolyploid

Porella platyphylla andP. cordaeana are genetically well differentiated in Europe (I = 0.486). Isozyme data from 156 European colonies reveal thatP. baueri is polyploid and could be an interspecific hybrid between these two species. It is characterized by large lobe cells, toothed margins of the female bracts, and ciliate perianth mouth. Correspondence analysis using morphological characters revealed that its colonies cluster into two geographicallý separated groups, similar to one or other of the putative parental species. These results agree with the genetic analysis, that also revealed two main allelic associations, each with a distinct geographical range. We hypothesize that this pattern was produced by introgression subsequent to hybridization, or by gene silencing. The discovery of this new, allopolyploid species suggests that hybridization is an important evolutionary process in liverworts.     

Parallel speciation or long‐distance dispersal? Lessons from seaweeds (Fucus) in the Baltic Sea

Parallel evolution has been invoked as a forceful mechanism of ecotype and species formation in many animal taxa. However, parallelism may be difficult to separate from recently monophyletically diverged species that are likely to show complex genetic relationships as a result of considerable shared ancestral variation and secondary hybridization in local areas. Thus, species' degrees of reproductive isolation, barriers to dispersal and, in particular, limited capacities for long‐distance dispersal will affect demographical structures underlying mechanisms of divergent evolution. Here, we used nine microsatellite DNA markers to study intra‐ and interspecific genetic diversity of two recently diverged species of brown macroalgae, Fucus radicans (L. Bergström & L. Kautsky) and F. vesiculosus (Linnaeus), in the Baltic Sea. We further performed biophysical modelling to identify likely connectivity patterns influencing the species' genetic structures. For each species, we found intraspecific contrasting patterns of clonality incidence and population structure. In addition, strong genetic differentiation between the two species within each locality supported the existence of two distinct evolutionary lineages (F_ST = 0.15–0.41). However, overall genetic clustering analyses across both species' populations revealed that all populations from one region (Estonia) were more genetically similar to each other than to their own taxon from the other two regions (Sweden and Finland). Our data support a hypothesis of parallel speciation. Alternatively, Estonia may be the ancestral source of both species, but is presently isolated by oceanographic barriers to dispersal. Thus, a limited gene flow in combination with genetic drift could have shaped the seemingly parallel structure.     

Hybridization between two sympatrically breeding species of fur seal at Iles Crozet revealed by genetic analysis

Introgressive hybridization occurs when closely related taxa overlap in distribution and is often associated with historically isolated populations coming into contact as a result of anthropogenic disturbance. There is evolutionary and conservation interest in detecting hybridization to determine its implications on future species composition, especially for threatened and recovering taxa such as subantarctic (Arctocephalus tropicalis) and Antarctic (A. gazella) fur seals, which were driven to the brink of extinction by human exploitation. Hybridization between these species has been reported at two locations and they breed sympatrically at a third site, Iles Crozet. While hybrid individuals have previously been identified based on phenotype, individuals can be difficult to classify based on these characteristics alone. Genotypic hybrid identification has been successful in several species, including fur seals. In this study we conducted an assignment test using microsatellite data to identify hybrids and to measure the frequency of hybridization at Iles Crozet. Samples were collected from 372 individuals and screened with 6 polymorphic microsatellite markers. MtDNA genotypes were also determined for individuals identified as hybrids or backcrosses based on microsatellite genotype. Phenotype, microsatellite and mtDNA genotype were then compared in order to identify hybrids. The results indicate that 1% of the population have hybrid genotypes and at a minimum, 2.4% of the population are backcrossed to parental species. We found that the two species are genetically distinct from one another and given the low rate of hybridization it is unlikely that they will fuse. These results suggest that there is a mechanism for species recognition that acts as a barrier to hybridization. It therefore seems unlikely that fur seals are threatened by significant introgression. Further investigation of fur seal mating systems would provide valuable insight into the mechanisms governing hybridization and species recognition in mate choice.     

Phenotypic and genetic diversity in aposematic Malagasy poison frogs (genus Mantella)

Intraspecific color variation has long fascinated evolutionary biologists. In species with bright warning coloration, phenotypic diversity is particularly compelling because many factors, including natural and sexual selection, contribute to intraspecific variation. To better understand the causes of dramatic phenotypic variation in Malagasy poison frogs, we quantified genetic structure and color and pattern variation across three closely related species, Mantella aurantiaca, Mantella crocea, and Mantella milotympanum. Although our restriction site‐associated DNA (RAD) sequencing approach identified clear genetic clusters, they do not align with current species designations, which has important conservation implications for these imperiled frogs. Moreover, our results suggest that levels of intraspecific color variation within this group have been overestimated, while species diversity has been underestimated. Within major genetic clusters, we observed distinct patterns of variation including: populations that are phenotypically similar yet genetically distinct, populations where phenotypic and genetic breaks coincide, and populations that are genetically similar but have high levels of within‐population phenotypic variation. We also detected admixture between two of the major genetic clusters. Our study suggests that several mechanisms—including hybridization, selection, and drift—are contributing to phenotypic diversity. Ultimately, our work underscores the need for a reevaluation of how polymorphic and polytypic populations and species are classified, especially in aposematic organisms.     

Hybridization and the evolution of invasiveness in plants and other organisms

Less than a decade ago, we proposed that hybridization could serve as a stimulus for the evolution of invasiveness in plants (Ellstrand and Schierenbeck Proc Nat Acad Sci USA 97:7043–7050, 2000). A substantial amount of research has taken place on that topic since the publication of that paper, stimulating the symposium that makes up this special issue. Here we present an update of this emergent field, based both on the papers in this volume and on the relevant literature. We reevaluate the lists that we presented in our earlier paper of reports in which hybridization has preceded the evolution of invasiveness. We discard a few cases that were found to be in error, published only as abstracts, or based on personal communication. Then we augment the list from examples in this volume and a supplementary literature search. Despite the omissions, the total number of cases has increased. Many have been strengthened. We add a list of cases in which there has been evidence that intra-taxon hybridization has preceded the evolution of invasiveness. We also provide a number of examples from organisms other than plants. We consider how our examples suggest mechanisms whereby hybridization may act to stimulate the evolution of invasiveness. Hybridization does not represent the only evolutionary pathway to invasiveness, but it is one that can explain why the appearance of invasiveness often involves a long lag time and/or multiple introductions of exotics.     

Genetic relationships between peanut and wild species ofArachis sect.Arachis (Fabaceae): Evidence from RAPDs

Twenty-six accessions of wildArachis species and domesticated peanuts,A. hypogaea, introduced from South America were analyzed for random amplified polymorphic DNA (RAPD). The objective of the study was to investigate inter- and intraspecific variation and affinities among species of sect.Arachis which have been proposed as possible progenitors for the domesticated peanut. Ten primers resolved 132 DNA bands which were useful for separating species and accessions. The most variation was observed among accessions ofA. cardenasii andA. glandulifera whereas the least amount of variation was observed inA. hypogaea andA. monticola. The two tetraploid species could not be separated by using RAPDs.Arachis duranensis was most closely related to the domesticated peanut and is believed to be the donor of the A genome. The data indicated thatA. batizocoi, a species previously hypothesized to contribute the B genome toA. hypogaea, was not involved in its evolution. The investigation showed that RAPDs can be used to analyze both inter- and intraspecific variation in peanut species. Southern hybridization of RAPD probes to blots containing RAPD of theArachis species provided information on genomic relationships and revealed the repetitive nature of the amplified DNA.     

Evolution of mating systems inLycopersicon hirsutum as deduced from genetic variation in electrophoretic and morphological characters

Populations in the central part of the distribution are mostly self-incompatible and tend to be highly variable for allozymic and morphological characters; those in the north and south limits are entirely self-compatible and tend to be genetically highly uniform. Gradations in variability are observed in the intermediate regions. Flower size tends to diminish in the peripheral areas. The extensive differences in genotype observed between the north and south marginal populations are not compatible with the concept of a single origin of self-compatibility, but suggest, along with other evidence, that the substitution of different alleles resulted from differentiation in the marginal areas from older, self-incompatible stocks of the central region. The conclusions regarding patterns of genetic variation and nature of evolution of mating systems inL. hirsutum conform to a remarkable extent with those reached previously forL. pimpinellifolium, a species that is distinct in morphology and ecological preferences yet has a similar latitudinal distribution.     

A Comparison of Hybridization between Mottled Ducks (<Emphasis Type="Italic">Anas fulvigula</Emphasis>)and Mallards (<Emphasis Type="Italic">A. platyrhynchos</Emphasis>) in Florida and South Carolina using Microsatellite DNA Analysis

Interspecific hybridization has been implicated in population declines for some waterfowl species within the mallard complex, and hybridization with mallards (Anas platyrhynchos) is currently considered the largest threat to mottled ducks (A. fulvigula), one North American member of that complex. We assessed genetic variation among 225 mottled ducks and mallards using five microsatellite loci, and detected significant overall differences between these species within two geographic areas. We characterized hybridization in Florida, where mottled ducks are endemic and mallards are beginning to appear on the breeding grounds, and in South Carolina, where mottled ducks were introduced outside their native range. We used Bayesian genetic mixture analysis in an attempt to distinguish between these closely related species. In Florida, we detected two distinct genetic groups, and 10.9% of our samples from Florida mottled ducks were inferred to have been hybrids. In contrast only 3.4% of Florida mallards were inferred to have been hybrids, suggesting asymmetric hybridization. Populations from different geographic areas within Florida exhibited hybridization rates ranging from 0% to 24%. These data indicate a genetic component would be appropriate in actively managing interspecific hybridization in Florida mottled ducks. In contrast, South Carolina mottled ducks and mallards cannot be differentiated.     

Phylogenetic species delimitation in ectomycorrhizal fungi and implications for barcoding: the case of the Tricholoma scalpturatum complex (Basidiomycota)

Population studies have revealed that the fungal ectomycorrhizal morphospecies Tricholoma scalpturatum consists of at least two genetically distinct groups that occur sympatrically in several geographical areas. This discovery prompted us to examine species boundaries and relationships between members formerly assigned to T. scalpturatum and allied taxa using phylogenetic analyses. Sequence data were obtained from three nuclear DNA regions [internal transcribed spacer (ITS), gpd and tef], from 101 carpophores collected over a large geographical range in Western Europe, and some reference sequences from public databases. The ITS was also tested for its applicability as DNA barcode for species delimitation. Four highly supported phylogenetic clades were detected. The two previously detected genetic groups of T. scalpturatum were assigned to the phylospecies Tricholoma argyraceum and T. scalpturatum. The two remaining clades were referred to as Tricholoma cingulatum and Tricholoma inocybeoides. Unexpectedly, T. cingulatum showed an accelerated rate of evolution that we attributed to narrow host specialization. This study also reveals recombinant ITS sequences in T. inocybeoides, suggesting a hybrid origin. The ITS was a useful tool for the determination of species boundaries: the mean value of intraspecific genetic distances in the entire ITS region (including 5.8S rDNA) was <0.2%, whereas interspecific divergence estimates ranged from 1.78% to 4.22%. Apart from giving insights into the evolution of the T. scalpturatum complex, this study contributes to the establishment of a library of taxonomically verified voucher specimens, an a posteriori correlation between phenotype and genotype, and DNA barcoding of ectomycorrhizal fungi.     

Molecular investigations of pathogenesis-related Bet v 1 homologues in Passiflora (Passifloraceae)

The major birch pollen allergen, Bet v 1, responsible for allergic reactions in many areas of the world, is homologous to a large number of pathogenesis-related proteins (PRs), identified as PR10. As part of a long-range investigation of these types of proteins and of evolution in Passiflora,DNA sequences from eight Bet v 1 homologue isoforms were obtained from five species of this genus in Brazil, and their sequences compared among themselves and with 30 others from 8 different species, classified in different taxonomic units. The objective was a first characterization of these PRs in wild passionflowers, and their use for evolutionary and applied investigations. High interspecific, but low intraspecific variability was observed, as expected from multigenic families subjected to concerted evolution. The relationships obtained both within Passiflora and between it and seven other genera probably best reflect functional similarities than evolutionary history.     

Intraspecific genetic analysis of the summer tanager Piranga rubra: implications for species limits and conservation

The summer tanager Piranga rubra is a Neotropical migrant that has experienced noted declines in the southwestern United States caused by extensive habitat loss of native riparian woodlands. This species is composed of two morphologically and behaviorally distinct taxa that traditionally have been recognized as subspecies, each occupying unique habitats in the southern part of North America. Genetic analyses of intraspecific variation are important in studies of threatened or endangered species because they can indicate whether smaller management units exist below the species level and they also provide estimates of within population variability. Using a mitochondrial DNA marker, the intraspecific genetic variation of this species is explored to determine whether the morphologically and behaviorally distinct subspecies are also genetically unique. By using traditional phylogenetic methods and building haplotype networks, results from this study indicate that the subspecies represent two phylogenetic species and should be managed as separate units. In addition, the level of gene flow among geographically isolated populations of the western subspecies is explored using Nested Clade Phylogeographic Analysis and population genetic tests. These analyses show that populations are genetically diverse and that haplotypes are shared across populations. Newly colonized populations are as diverse as older populations. This suggests that as habitat degrades in traditional breeding areas of the summer tanager, if suitable habitat elsewhere becomes available for new populations, these new colonies should be genetically diverse.     

Intraspecific sterility barrier confirms that introduction of Sphaerotrichia divaricata (Phaeophyceae,Chordariales) into the Mediterranean was from Japan

Crossing studies revealed an intraspecific sterility barrier on the level of zygote formation between Japanese Sphaerotrichia divaricata and isolates of the same species from the Northeast Pacific and the North Atlantic. Because no consistent morphological differences exist between sporophytes from Japan and other areas, we propose not to distinguish the intersterile populations as different species. Japanese Sphaerotrichia and isolates from a recently detected population in the Étang de Thau, French Mediterranean coast, are interfertile. The crossing studies support the assumption that this Mediterranean population is a recent introduction from Japan.     

Primate numts and reticulate evolution of capped and golden leaf monkeys (Primates: Colobinae)

A recent phylogenetic study of langurs and leaf monkeys of South Asia suggested a reticulate evolution of capped and golden leaf monkeys through ancient hybridization between Semnopithecus and Trachypithecus. To test this hybridization scenario, I analysed nuclear copies of the mitochondrial cytochrome b gene (numts) from capped, golden and Phayre’s leaf monkeys. These numts were aligned with mitochondrial cytochrome b sequences of various species belonging to the genera Semnopithecus and Trachypithecus. In the phylogenetic tree derived from this alignment, the numts fell into three distinct clades (A, B and C) suggesting three independent integration events. Clade A was basal to Semnopithecus, and clades B and C were basal to Trachypithecus. Among the numts in clades A and C were sequences derived from species not represented in their respective sister mitochondrial groups. This unusual placement of certain numts is taken as additional support for the hybridization scenario. Based on the molecular dating of these integration events, hybridization is estimated to have occurred around 7.1 to 3.4 million years ago. Capped and golden leaf monkeys might have to be assigned to a new genus to reconcile their unique evolutionary history. Additionally, northeast India appears to be a ‘hot spot’ for lineages that might have evolved through reticulate evolution.     

Taxonomic and evolutionary implications of the Endosperm Balance Number hypothesis in potatoes

Summary The Endosperm Balance Number (EBN) hypothesis can explain to a considerable degree the crossability between tuber-bearing Solanum species. It has been shown to be genetically controlled and is dosage dependent. There is a good correlation between EBN and the postulated evolution and present taxonomy of potatoes. The primitive white stellate-flowered species from Mexico are 1EBN, and this condition is also found in species from South America with flowers of the same colour and shape. The evolution of a rotate corolla seems to be correlated with 2EBN. It is postulated that the 2EBN state arose as a reproductive isolating mechanism in South America. The taxonomic and evolutionary implications of the EBN hypothesis are discussed.     

Evolution of enhanced reproduction in the hybrid-derived invasive, California wild radish (Raphanus sativus)

Evolution is receiving increased attention as a potentially important factor in invasions. For example, hybridization may have stimulated the evolution of invasiveness in several well-known plant pests. However, the mechanism for success of such hybrid-derived lineages remains unknown in the majority of the cases studied. Here we ask whether increased reproductive success (in terms of maternal fitness) has evolved in an invasive lineage with confirmed hybrid ancestry. We compare the relative fitness of the non-native, hybrid-derived California wild radish (Raphanus sativus) to that of its two progenitor species in field experiments at different sites and in different years. We found that California wild radish has high survivorship and produces more fruits per plant and more seeds per plant than either of its progenitors in several environments. Furthermore, populations of California wild radish display a strong genotype-by-environment interaction, indicating that maintenance of genetic and phenotypic diversity between populations may be responsible for the weed’s ability to invade a wide breadth of California habitats. Our results suggest that hybridization may contribute the evolution of enhanced invasiveness and, also, that by limiting the introduction and subsequent hybridization of congeners, we may be able to prevent the evolution of new invasive lineages.