Limited hybridization between Quercus lobata and Quercus douglasii (Fagaceae) in a mixed stand in central coastal California

Many oak species are interfertile, and morphological and genetic evidence for hybridization is widespread. Here we use DNA microsatellite markers to characterize hybridization between two closely related oak species in a mixed stand in central coastal California, Quercus lobata (valley oak) and Q. douglasii (blue oak) (Fagaceae). Genotypes from four microsatellite loci indicate that many alleles are shared between the two species. However, each species harbors unique alleles, and allele frequencies differ significantly. A Bayesian analysis of genetic structure in the stand identified two highly differentiated genetic clusters, essentially corresponding to species assignment based on morphology. Data from the four loci were sufficient to assign all 135 trees to one of the two species. In addition, five putative hybrid individuals having intermediate morphologies could be assigned genetically to one or the other species, and all but one had low probability of hybrid ancestry. Overally, only six (4.6%) trees showed >0.05 probability of hybrid ancestry, in all cases their probabilities for nonhybrid ancestry were substantially higher. We conclude that adult hybrids of Q. douglasii × Q. lobata are rare at this site and plasticity in morphological characters may lead to overestimates of hybridization among Quercus species.     

Variation in morphological traits in a recent hybrid zone between closely related Quercus liaotungensis and Q. mongolica (Fagaceae)

Aims Hybridization usually leads to gene introgression between related species in hybrid zones, associated with complex patterns of morphological variation. Nevertheless, previous studies have tended to ignore the effects of geographic variation in hybridization rates on species taxonomy. This study aims to investigate the variation of morphological traits between two sympatric and taxonomically confused oak species, Quercus liaotungensis and Q. mongolica, and reveal the effects of hybridization rates on morphological traits and the taxonomic boundary.Methods We used seven microsatellite loci to evaluate species status and measured 15 morphological traits in 26 trees in the recent hybrid zone between Q. liaotungensis and Q. mongolica, and we characterized the differences between the two oak species and their hybrids for the investigated traits.Important findings Molecular analyses indicated that 74% of 78 sampled maternal trees were hybrids between Q. liaotungensis and Q. mongolica although the observed morphological variation suggested that they had remained distinct species. Across all of the differentiated leaf and reproductive traits, the hybrids expressed patterns similar to Q. liaotungensis, which may suggest dominant expression of parental characters. These results are consistent with our expectation that hybrids will be difficult to distinguish from parental species in a recent hybrid zone.     

Community structure of canopy arthropods associated to Quercus crassifolia×Quercus crassipes complex

Quercus crassifolia and Q. crassipes are dominant species in temperate forests of central Mexico and hybridize between each other when they occur in sympatry. Oak canopies contain a considerable portion of arthropod diversity and the hybrid zones can provide new habitats to epiphyte fauna. We tested if the establishment of hybrids in contact zones with their parental hosts increases the species diversity of canopy arthropods assuming that hybrid trees constitute new genotypes of potential new habitats to small organisms. We examined the effect of hybridization on some community structure parameters (diversity, composition, similarity and density of arthropod fauna) of canopy arthropods compared to their parental species in a hybrid zone located in central Mexico. We employed 17 leaf morphological traits and six diagnostic RAPD primers to identify parental and hybrid plants. The RAPDs marker showed unidirectional introgression towards Q. crassifolia, and were detected hybrid (F₁), backcrosses and introgression individual trees. In total, 30 oak canopies were fogged during rainy and dry season. We recognized 532 taxa of arthropods belonging to 22 orders associated with tree canopies. The taxonomic status of host‐trees may be an important factor in the arthropod community structure and that seasonality (dry and rainy) is not a factor that could modify their organization. Trees of Q. crassipes registered the highest densities of arthropod fauna followed by hybrid hosts (F₁); trees originated by backcrosses towards Q. crassifolia registered a significant less arthropod density than F₁ hybrids; and trees of Q. crassifolia had the lowest density. Hybrid plants and Q. crassipes individuals had higher diversity (H′) of arthropods than Q. crassifolia plants. Hybrid plants had also more rare species in both seasons in comparison with parental species. This study suggests that hybrid oaks act as a center of biodiversity by accumulating arthropods of both parental and different species including a considerable number of rare species.     

Coincidence of small-scale spatial discontinuities in leaf morphology and nuclear microsatellite variation of Quercus petraea and Q. robur in a mixed forest

BACKGROUND AND AIMS: The taxon complex comprising Quercus petraea and Q. robur shows distinct morphologies and ecological preferences, but mostly low differentiation in various types of molecular markers at a broad spatial range. Local, spatially explicit analyses may reveal patterns induced by microevolutionary processes operating mainly over short distances. However, no attempts have been made to date to explore the potential of spatial analyses combining morphological and genetic data of these oaks. METHODS: A mixed oak stand was studied to elucidate the small-scale population genetic structure. All adult individuals were classified and putative hybrids were identified using multivariate discrimination analysis of leaf morphological characters. Likewise, all trees were genotyped with five nuclear microsatellites, and a Bayesian assignment method was applied based on maximum likelihood of multilocus genotypes for taxon and putative hybrid classification. KEY RESULTS: Multivariate analyses of leaf morphological data recognized two groups with few individuals as putative hybrids. These groups were significantly differentiated at the five microsatellites, and genetic taxon assignment coincided well with morphological classification. Furthermore, most putative hybrids were assigned to the taxon found in their spatial neighbourhood. When grouping trees into clusters according to their spatial positions, these clusters were clearly dominated by one taxon. Discontinuities in morphological and genetic distance matrices among these clusters showed high congruence. CONCLUSIONS: The spatial-genetic analyses and the available literature led to the assumption that reproductive barriers, assortative mating, limited seed dispersal and microsite-induced selection in favour of the locally adapted taxon at the juvenile stage may reinforce taxon-specific spatial aggregation that fosters species separation. Thus, the results tend to support the hypothesis that Q. petraea and Q. robur are distinct taxa which share a recent common ancestry. Occasional hybrids are rarely found in adults owing to selection during establishment of juveniles.     

Comparative analysis of micromorphological characters in two distantly related Mexican oaks, Quercus conzattii and Q. eduardii (Fagaceae), and their hybrids

Interspecific hybridization occurs with high frequency in the genusQuercus, but few studies have analyzed and compared micromorphological characters in putative parental species and their hybrids.Quercus eduardii andQ. conzattii are two Mexican black oak species that, although distantly related, have formed at least one population of hybrid origin, where individuals with intermediate macromorphology are present. The purpose of this investigation was to analyze the degree of differentiation in micromorphological characters between the two species and to assess the expression of these characters in individuals with intermediate macromorphology. Foliar trichomes, epicuticular waxes, stomata, and pollen grains, were examined using scanning electron microscopy (SEM) in the three types of individuals (Q. conzatti, Q. eduardii, and intermediates). Trichome density was quantified with light microscopy. Types of trichomes present, length of trichome arms, types of epicuticular wax on the leaf surfaces, and the position of stomata with respect to the foliar surface were characters useful to differentiate betweenQ. conzattii andQ. eduardii. Plants with intermediate macromorphology displayed a pattern of micromorphological characters that were identical to one parental species (Q. conzattii), or extreme or novel relative to both species.     

STUDIES IN THE QUERCUS UNDULATA COMPLEX. III. THE CONTRIBUTION OF Q. ARIZONICA

Tucker, J. M. (U. California, Davis.) Studies in the Quercus undulata complex. III. The contribution of Q. arizonica . Amer. Jour. Bot. 50(7): 699–708. Illus. 1903.—Of the 7 oak species involved in the Quercus undulata complex, Q. arizonica contributes the least of all. The latter has hybridized with Q. gambelii at a few widely separated localities (listed in Table 1) in central Arizona, and northern Sonora and Chihuahua, Mexico. The putative hybrids (identifiable as Quercus undulata) occur as isolated individuals with the parental species. The parents, although regionally sympatric, are usually ecologically isolated. They differ in a number of morphological characters, 6 of which were analyzed in detail. The resulting data, presented as pietorialized scatter diagrams, demonstrate that the putative hybrids are intermediate, in general. This is taken as evidence of their hybrid nature. Factors limiting hybridization in oaks are discussed. It is speculated that hybridization between Q. arizonica and Q. gambelii has occurred since the postglacial hypsithermal interval.     

Hybridization increases canopy arthropod diversity in the <Emphasis Type="Italic">Quercus affinis</Emphasis> × <Emphasis Type="Italic">Quercus laurina</Emphasis> complex

Understanding the factors that influence the diversity and composition of arthropod communities is a major topic in ecology. Canopy arthropod communities are a major constituent of biodiversity and show great variation in time and space according to different factors. Recently, genetic variation within tree species has attracted attention as a significant factor determining the diversity and composition of canopy arthropod communities. A major source of genetic and phenotypic novelty in plant species is interspecific hybridization, and therefore it is of interest to evaluate how this process affects the communities of associated organisms. In this study, we used microsatellite markers and geometric morphometry of leaf shape to analyze genetic and morphological variation in 45 individuals in a local hybrid zone between the oaks Quercus affinis and Q. laurina in Mexico. Individual trees were assigned to one of the parental species or to the hybrid category. The percentage of leaf area removed by herbivores was quantified in each individual and the canopies of five individuals of each categeory (two parental species and hybrids) was fogged with insecticide to assess the diversity and composition of arthropod communities. Results indicated that hybrid trees experience higher levels of herbivory than parental species and also sustain a higher abundance and richness of canopy arthropods. In general, our study supports the “hybrid susceptibility hypothesis” that predicts a higher incidence of associated arthropods on hybrid plants than in their parental species as result of the disruption of co-adapted gene complexes associated to resistance traits.     

Molecular evidence for hybrid origin of Quercus crenata Lam. (Fagaceae) from Q. cerris L. and Q. suber L.

Abstract

Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeats (ISSR) markers were employed to examine samples from Quercus cerris, Q. suber and Q. crenata in order to test the hypothesis of the hybrid origin of Q. crenata from Q. cerris and Q. suber in a part of its distribution area where the two putative parents do not overlap at present. Leaves from 21 Q. crenata and 37 Q. cerris individual trees were collected at natural sites in northern Italy, where Q. suber is currently lacking; 21 Q. suber and six Q. crenata plants from central Italy were also analysed. Results from Unweighted Pair Group Method with Arithmetic mean (UPGMA) analysis and principal component analysis (PCA) implied that exchange of neutral markers has been considerable between the three species, while differences in morphological characters have remained comparatively stable. The Mantel test indicated low correlation between RAPD- and ISSR-based similarity matrices, showing that the two screening techniques reveal unrelated estimates of genetic relationships. Hybrid indices computed for both markers displayed an intermediate position of Q. crenata individuals between the two putative parents shifted toward Q. cerris. Results from the present study corroborate the hypothesis of a hybrid origin for Q. crenata occurring in northern Italy, and suggest asymmetrical backcrossing with Q. cerris acting as the recurrent parent.     

Asymmetrical introgression between two Morus species (M. alba, M. rubra) that differ in abundance

Asymmetrical introgression is an expected genetic consequence of hybridization when parental taxa differ in abundance; however, evidence for such effects in small populations is scarce. To test this prediction, we estimated the magnitude and direction of hybridization between red mulberry (Morus rubra L.), an endangered species in Canada, and the introduced and more abundant white mulberry (Morus alba L.) using nuclear (randomly amplified polymorphic DNA) and cytoplasmic (chloroplast DNA sequence) markers. Parentage of 184 trees (n = 42 using cpDNA) from four sympatric populations was estimated using a hybrid index and related to six morphological characters and population frequencies of the parental classes. Overall, the frequency of nuclear hybrids was 53.7% (n = 99) and ranged from 43% to 67% among populations. The parental and hybrid taxa differed with respect to all of the morphological traits. Sixty-seven percent of all hybrids contained more nuclear markers from M. alba than M. rubra (hybrid index x = 0.46); among populations, the degree of M. alba bias was correlated with the frequency of M. alba. In addition, the majority of hybrids (68%) contained the chloroplast genome of white mulberry. These results suggest that introgression is bidirectional but asymmetrical and is related, in part, to the relative frequency of parental taxa.     

Morphological and RAPD analysis of hybridization between Quercus affinis and Q. laurina (fagaceae), two Mexican red oaks

Quercus affinis and Q. laurina are two closely related Mexican red oaks with partially overlapping distributions. Within the area of overlap, there are localities where morphological intergradation occurs. A previous hypothesis explained this pattern as a result of secondary contact between the two species, followed by hybridization and introgression. This possibility was analyzed here by examining foliar and genetic variation in 16 localities situated along a macrogeographic gradient, which included morphologically representative populations of both species and populations from within the area of overlap. Maximum-likelihood hybrid index scores calculated from nine semi-diagnostic RAPD markers indicated a shift in the genetic composition of populations from one species to the other along the macrogeographic gradient, with genetically intermediate populations situated in the area of overlap. Foliar variation followed a partially congruent pattern, but Q. laurina-like morphology predominated in some of the genetically intermediate populations. There were several instances of correlated frequency changeovers of single RAPD markers and morphological characters along the macrogeographic gradient and a few cases of markedly parallel patterns between markers. The results were interpreted as consistent with a hypothesis of secondary contact between the two oak species that has resulted in some differential introgression among markers.     

Geographic variation in the structure of oak hybrid zones provides insights into the dynamics of speciation

Studying geographic variation in the rate of hybridization between closely related species could provide a useful window on the evolution of reproductive isolation. Reinforcement theory predicts greater prezygotic isolation in areas of prolonged contact between recently diverged species than in areas of recent contact, which implies that old contact zones would be dominated by parental phenotypes with few hybrids (bimodal hybrid zones), whereas recent contact zones would be characterized by hybrid swarms (unimodal hybrid zones). Here, we investigate how the hybrid zones of two closely related Chinese oaks, Quercus mongolica and Q. liaotungensis, are structured geographically using both nuclear and chloroplast markers. We found that populations of Q. liaotungensis located around the Changbai Mountains in Northeast China, an inferred glacial refugium, were introgressed by genes from Q. mongolica, suggesting historical contact between the two species in this region. However, these introgressed populations form sharp bimodal hybrid zones with Q. mongolica. In contrast, populations of Q. liaotungensis located in North China, which show no sign of ancient introgression with Q. mongolica, form unimodal hybrid zones with Q. mongolica. These results are consistent with the hypothesis that selection against hybrids has had sufficient time to reinforce the reproductive barriers between Q. liaotungensis and Q. mongolica in Northeast China but not in North China.     

Transgressive character expression in hybrid zones between the native invasives Tithonia tubaeformis and Tithonia rotundifolia (Asteraceae) in Mexico

Natural hybridization frequently promotes gene introgression among closely related species in sympatric populations, producing complex patterns of morphological variation. Therefore, a detailed understanding of the dynamics of interspecific gene flow and its morphological patterns is of widespread interest. We tested if introgressive hybridization promotes an increase in transgressive characters in comparison with the parental species. A sunflower species complex occurring in Mexico formed by two native invasive species, Tithonia tubaeformis and Tithonia rotundifolia, was analyzed using 46 morphological characters (leaf, flower and fruit) in five hybrid zones (N = 150 individuals) and two pure sites for each parental species (N = 80 individuals). In general, T. tubaeformis differed significantly from T. rotundifolia in all the examined characters, except six foliar and one inflorescence character. Morphological characters support the hypothesis of hybridization in this complex, even though both species remain morphological distinct in mixed stands. Individual hybrids appear to be a mosaic of parent-like (24.8 % of traits), intermediate (26.1 %) and transgressive (37.8 %) phenotypes (the remaining 11.3 % of the traits did not differ significantly from both parental species). Our results suggest that individuals from the same parental species were more similar among themselves than to putative hybrids, indicating occasional hybridization with segregation in hybrid types or backcrossing to parents. Evidence indicates a unidirectional pattern of gene flow toward T. rotundifolia.     

Effect of hybridization of the Quercus crassifolia×Quercus crassipes complex on the community structure of endophagous insects

In a previous study, we showed that the geographic proximity of hybrid plants to the allopatric areas of parental species increases their morphological and genetic similarity with them. In the present work, we explored whether the endophagous fauna of hybrid plants show the same pattern. We studied the canopy species richness, diversity and composition of leaf-mining moths (Lepidoptera: Tischeridae, Citheraniidae) and gall-forming wasps (Hymenoptera: Cynipidae) associated with two species of red oaks (Quercus crassifolia and Quercus crassipes) and their interspecific hybrid (Quercus×dysophylla Benth pro sp.) in seven hybrid zones in central Mexico, during four seasons in 2 years. The study was conducted on 194 oak trees with known genetic status [identified by leaf morphology and molecular markers (random amplified polymorphic DNAs)], and the results indicate a bidirectional pattern of gene flow. Hybrid plants supported intermediate levels of infestation of gall-forming and leaf-mining insects compared to their putative parental species. The infestation level of leaf-mining insects varied significantly following the pattern: Q. crassifolia>hybrids>Q. crassipes, whereas the gall-forming insects showed an inverse pattern. A negative and significant relationship was found between these two types of insect guilds in each host taxa, when the infestation percentage was evaluated. It was found that 31.5% (n=11) of the endophagous insects were specific to Q. crassipes, 22.9% (n=8) to Q. crassifolia, and 8.6% (n=3) to hybrid individuals. The hybrid bridge hypothesis was supported in the case of 25.7% (n=9) of insects, which suggests that the presence of a hybrid intermediary plant may favor a host herbivore shift from one plant species to another. Greater genetic diversity in a hybrid zone is associated with greater diversity in the endophagous community. The geographic proximity of hybrid plants to the allopatric site of a parental species increases their similarity in terms of endophagous insects and the Eje Neovolcánico acts as a corridor favoring this pattern. Electronic Supplementary Material Supplementary material is available for this article at     

STUDIES IN THE QUERCUS UNDULATA COMPLEX. II. THE CONTRIBUTION OF QUERCUS TURBINELLA

Tucker , J. M. (U. California, Davis), W. P. Cottam , and R. Drobnick . Studies in the Quercus undulata complex. II. The contribution of Quercus turbinella. Amer. Jour. Bot. 48(4): 329–339. Illus. 1961.—Quercus undulata has been interpreted as a hybrid complex involving Q. gambelii and several other species, including Q. turbinella (Tucker, 1961). In the present paper, the total distribution of the hybrids between Q. gambelii and Q. turbinella is given. Lacking direct genetic evidence, proof of hybridity is sought in a demonstration of the morphological intermediacy of these putative hybrids. Population samples of both parental species, other samples containing hybrids, and numerous individual hybrids, are analyzed on the basis of 6 differences between the parental species. The data obtained, presented in the form of pictorialized scatter diagrams, clearly show the general intermediacy of the hybrids. Of the various binomials that have been applied to forms in the Q. undulata complex, Quercus pauciloba Rydb. applies to this hybrid. The appropriate change in status (Quercus × pauciloba Rydb.) is made.     

Patterns of hybridization and introgression between invasive Ulmus pumila (Ulmaceae) and native U. rubra

Natural hybridization between introduced species and their native congeners occurs frequently and can create serious conservation concerns. Ulmus pumila (Siberian elm) is an introduced Asian elm species that has naturalized in the United States and is now considered invasive in 41 states. Red elm (U. rubra), a native to the eastern United States, often occurs in sympatry with Siberian elm, and the two species are thought to hybridize. Here, we genetically characterized reference populations of the two elm species to identify species-specific microsatellite alleles. These markers were used to classify individuals in putative hybrid zones as parental species or hybrids, assess the extent of hybridization, and track patterns of introgression. We identified nine U. rubra, 32 U. pumila, and 51 hybrid individuals in our hybrid zones. Of the 51 hybrids, 35 were classified as first-generation hybrids and 16 as backcrosses. The majority of the backcrosses (88%) were introgressed toward U. pumila. Our classification of genotypes was consistent whether we used manual classification, principal coordinate analyses or Bayesian clustering. We observed greater genetic diversity and new combination of alleles in the hybrids. Our study indicates widespread hybridization between U. pumila and U. rubra and an asymmetric pattern of introgression toward U. pumila.     

Morphological and molecular evidence of natural hybridization in Shorea (Dipterocarpaceae)

Shorea (Dipterocarpaceae) is a large genus in which many closely related species often grow together in Southeast Asian lowland tropical rain forests. Many Shorea species share common pollinators, and earlier studies suggested occurrence of interspecific hybridization and introgression. Here, we show morphological and molecular evidence of hybridization between Shorea species. In the census of all the trees of Shorea curtisii, Shorea leprosula, and Shorea parvifolia (>30 cm dbh) within the 164-ha area of Bukit Timah Nature Reserve in Singapore, we found 21 morphologically recognizable hybrid individuals. All of the putative hybrids could be distinguished obviously from the parental species on the basis of vegetative characters. Population genetic analysis of DNA sequences of two nuclear (GapC and PgiC) and chloroplast (trnL-trnF) regions demonstrated that each of the three species had several species-specific mutations. The nuclear sequences of the putative hybrids were heterozygote at all the species-specific sites between two parental species. Hybrid between S. curtisii and S. leprosula was found most, while S. curtisii × S. parvifolia and S. leprosula × S. parvifolia hybrids were also found. Almost no shared polymorphism between populations of the parental species suggests rarity of introgression. The study indicated that natural hybridization between sympatric Shorea species should not be uncommon, but all of the hybrid individuals were F₁, and the post-F₁ hybrids were considerably rare.     

Identification of hybrids betweenQuercus petraea andQ. robur (Fagaceae): Results obtained with RAPD markers confirm allozyme studies based on theGot-2 locus

RAPDs were employed as genetic markers to detect interspecific hybridization between the closely related oak speciesQuercus robur andQ. petraea. Fourteen primers were used in order to check the genetic status (pure or hybrid) of individuals classified morphologically. Among the 147 PCR fragments obtained 11 appear to be species-specific. In the phenotypically intermediate individuals different combinations of these species-specific bands were obtained. The patterns in these putative hybrids were not additive, which may be either the result of repeated backcrossing and introgression between the two species or of heterozygosity within the parental species. The results of the RAPD study are consistent with morphological analyses and allozyme data obtained for theGot-2 locus. Thus the RAPD markers used in this study may provide a powerful genetic tool for the identification of hybrids and the discrimination between the two pure species.     

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.     

Interspecific hybridization in natural populations ofCyrtandra (Gesneriaceae) on the Hawaiian Islands: Evidence from RAPD markers

Interspecific hybridization among Hawaiian species ofCyrtandra (Gesneriaceae) was investigated using randomly amplified polymorphic DNA (RAPD) markers. Thirty-three different primers were used to investigate interspecific hybridization for 17 different putative hybrids based on morphological intermediacy and sympatry with putative parental species. RAPD data provided evidence for the hybrid origin of all putative hybrid taxa examined in this analysis. However, the patterns in the hybrid taxa were not found to be completely additive of the patterns found in the parental species. Markers missing in the hybrid taxa can be attributed to polymorphism in the populations of the parental species and the dominant nature of inheritance for RAPD markers. Unique markers found within hybrid taxa require further explanation but do not necessarily indicate that the taxa are not of hybrid origin. The implications suggest that these interspecific hybridization events had, and continue to have, an effect on the adaptive radiation and conservation biology ofCyrtandra.     

Morphometric variation in oaks of the Apostle Islands in Wisconsin: evidence of hybridization between Quercus rubra and Q. ellipsoidalis (Fagaceae)

The Apostle Islands in Lake Superior are populated by trees that are clearly related to Quercus rubra L. However, several islands have trees with morphological characteristics suggestive of hybridization with Q. ellipsoidalis Hill. Leaf specimens were collected from trees in five locations: the outermost island, an intermediate island, the nearest-shore island, the northeast shoreline, and an inland forest about 24 km from the shoreline. Seventeen landmarks were digitized for two to five leaves per tree. These landmarks were used to generate nine linear characters and three angles. These characters, along with the number of bristle tips per leaf, were used in various combinations for several principal component analyses. In addition, the landmark configurations were examined using rotational-fit methods. The patterns observed in both types of analysis indicate phenotypic variation coincident with a line connecting the two most distant sample sites. The location nearest the geographic center of this line is also nearest the center of the two-dimensional view of phenotypic variation. Trees at each site illustrate a distinctive pattern in the rotational-fit analyses, and patterns of co-variation in the morphometric characters are different for each site. The observed morphometric variation is consistent with the hypothesis that there is hybridization between these two species, most likely in the form of introgression from Q. ellipsoidalis into Q. rubra.