Introgression and persistence of cultivar alleles in wild carrot (Daucus carota) populations in the United States.

PREMISE: Cultivated species and their wild relatives often hybridize in the wild, and the hybrids can survive and reproduce in some environments. However, it is unclear whether cultivar alleles are permanently incorporated into the wild genomes or whether they are purged by natural selection. This question is key to accurately assessing the risk of escape and spread of cultivar genes into wild populations. METHODS: We used genomic data and population genomic methods to study hybridization and introgression between cultivated and wild carrot (Daucus carota) in the United States. We used single nucleotide polymorphisms (SNPs) obtained via genotyping by sequencing for 450 wild individuals from 29 wild georeferenced populations in seven states and 144 cultivars from the United States, Europe, and Asia. RESULTS: Cultivated and wild carrot formed two genetically differentiated groups, and evidence of crop-wild admixture was detected in several but not all wild carrot populations in the United States. Two regions were identified where cultivar alleles were present in wild carrots: California and Nantucket Island (Massachusetts). Surprisingly, there was no evidence of introgression in some populations with a long-known history of sympatry with the crop, suggesting that post-hybridization barriers might prevent introgression in some areas. CONCLUSIONS: Our results provide support for the introgression and long-term persistence of cultivar alleles in wild carrots populations. We thus anticipate that the release of genetically engineered (GE) cultivars would lead to the introduction and spread of GE alleles in wild carrot populations.

The decision-making process of leafcutting bees when selecting patches.

Change in land configuration is an important driver of pollinator decline. Understanding patch selection by bees in fragmented landscapes has therefore become imperative to guide the design of habitats that support pollinators and ensure their conservation. This is especially true for solitary bees that make up most bee species in the world. To elucidate the decision-making process of a solitary bee when selecting patches, we tested four models of patch attractiveness that differed in the role of patch size and isolation distance in the selection process. In these models, bees used both patch size and patch distance, only patch distance, or chose randomly among patches. When patch size was included, bees could estimate patch resources fully or partially. An experiment with a centre patch, surrounded by four peripheral patches of different sizes and distances from the centre, provided observed transition data to test against predictions derived from each of the models. The alfalfa leafcutting bee, Megachile rotundata, does not move randomly among patches. This bee uses both patch size and isolation distance when selecting a patch but can only evaluate patch resources partially. This knowledge can guide the design of habitats in fragmented landscapes to facilitate solitary bee conservation.

Population-specific responses of floral volatiles to abiotic factors in changing environments.

PREMISE: Shifts in abiotic factors can affect many plant traits, including floral volatiles. This study examined the response of floral volatiles to water availability and whether phenotypic plasticity to water availability differs among populations. It also investigated genetic differentiation in floral volatiles, determined the effect of temperature on phenotypic plasticity to water availability, and assessed temporal variation in floral scent emission between day and evening, since pollinator visitation differs at those times. METHODS: Rocky Mountain columbine plants (Aquilegia coerulea), started from seeds collected in three wild populations in Colorado, Utah, and Arizona, were grown under two water treatments in a greenhouse in Madison, Wisconsin, United States. One population was also grown under the two water treatments, at two temperatures. Air samples were collected from enclosed flowers using dynamic headspace methods and floral volatiles were identified and quantified by gas chromatography (GC) with mass spectrometry (MS). RESULTS: Emission of three floral volatiles increased in the wetter environment, indicating phenotypic plasticity. The response of six floral volatiles to water availability differed among populations, suggesting genetic differentiation in phenotypic plasticity. Five floral volatiles varied among populations, and emission of most floral volatiles was greater during the day. CONCLUSIONS: Phenotypic plasticity to water availability permits a quick response of floral volatiles in changing environments. The genetic differentiation in phenotypic plasticity suggests that phenotypic plasticity can evolve but complicates predictions of the effects of environmental changes on a plant and its pollinators.

Bee species visiting Medicago sativa differ in pollen deposition curves with consequences for gene flow.

PREMISE: Pollinator foraging behavior can influence pollen dispersal and gene flow. In many plant species a pollinator trips a flower by applying pressure to release its sexual organs. We propose that differences in tripping rate among grooming pollinators could generate distinct pollen deposition curves, the pattern of pollen deposition over successive flowers visited. This study compares the pollen deposition curves of two grooming pollinators, a social bumble bee and a solitary leafcutting bee, with distinct tripping rates on Medicago sativa flowers. We predict a steeper deposition curve for pollen moved by leafcutting bees, the pollinator with the higher tripping rate. METHODS: Medicago sativa plants carrying a gene (GUS) whose product is easily detected by staining, were used as pollen donors. After visiting the GUS plants, a bee was released on a linear array of conventional M. sativa plants. The number of GUS pollen grains deposited over successive flowers visited or over cumulative distances was examined. Distinct mixed effect Poisson regression models, illustrating different rates of decay in pollen deposition, were fitted to the pollen data for each bee species. RESULTS: Pollen decay was steeper for leafcutting bees relative to bumble bees for both models of flowers visited and cumulative distance, as predicted by their higher tripping rate. CONCLUSIONS: This is the first report of a difference in pollen deposition curves between two bee species, both grooming pollinators. Such differences could lead to distinct impacts of bee species on gene flow, genetic differentiation, introgression, and ultimately speciation.

Floral traits influencing plant attractiveness to three bee species: Consequences for plant reproductive success.

PREMISE OF THE STUDY: The ability to attract pollinators is crucial to plants that rely on insects for pollination. We contrasted the roles of floral display size and flower color in attracting three bee species and determined the relationships between plant attractiveness (number of pollinator visits) and seed set for each bee species. METHODS: We recorded pollinator visits to plants, measured plant traits, and quantified plant reproductive success. A zero-inflated Poisson regression model indicated plant traits associated with pollinator attraction. It identified traits that increased the number of bee visits and traits that increased the probability of a plant not receiving any visits. Different components of floral display size were examined and two models of flower color contrasted. Relationships between plant attractiveness and seed set were determined using regression analyses. KEY RESULTS: Plants with more racemes received more bee visits from all three bee species. Plants with few racemes were more likely not to receive any bee visits. The role of flower color varied with bee species and was influenced by the choice of the flower color model. Increasing bee visits increased seed set for all three bee species, with the steepest slope for leafcutting bees, followed by bumble bees, and finally honey bees. CONCLUSIONS: Floral display size influenced pollinator attraction more consistently than flower color. The same plant traits affected the probability of not being visited and the number of pollinator visits received. The impact of plant attractiveness on female reproductive success varied, together with pollinator effectiveness, by pollinator species.

The role of pollinators in maintaining variation in flower colour in the Rocky Mountain columbine, Aquilegia coerulea.

BACKGROUND AND AIMS: Flower colour varies within and among populations of the Rocky Mountain columbine, Aquilegia coerulea, in conjunction with the abundance of its two major pollinators, hawkmoths and bumble-bees. This study seeks to understand whether the choice of flower colour by these major pollinators can help explain the variation in flower colour observed in A. coerulea populations. METHODS: Dual choice assays and experimental arrays of blue and white flowers were used to determine the preference of hawkmoths and bumble-bees for flower colour. A test was made to determine whether a differential preference for flower colour, with bumble-bees preferring blue and hawkmoths white flowers, could explain the variation in flower colour. Whether a single pollinator could maintain a flower colour polymorphism was examined by testing to see if preference for a flower colour varied between day and dusk for hawkmoths and whether bumble-bees preferred novel or rare flower colour morphs. KEY RESULTS: Hawkmoths preferred blue flowers under both day and dusk light conditions. Naïve bumble-bees preferred blue flowers but quickly learned to forage randomly on the two colour morphs when similar rewards were presented in the flowers. Bees quickly learned to associate a flower colour with a pollen reward. Prior experience affected the choice of flower colour by bees, but they did not preferentially visit novel flower colours or rare or common colour morphs. CONCLUSIONS: Differences in flower colour preference between the two major pollinators could not explain the variation in flower colour observed in A. coerulea. The preference of hawkmoths for flower colour did not change between day and dusk, and bumble-bees did not prefer a novel or a rare flower colour morph. The data therefore suggest that factors other than pollinators may be more likely to affect the flower colour variation observed in A. coerulea.

Genetic structure and domestication of carrot (Daucus carota subsp. sativus) (Apiaceae).

PREMISE OF THE STUDY: Analyses of genetic structure and phylogenetic relationships illuminate the origin and domestication of modern crops. Despite being an important worldwide vegetable, the genetic structure and domestication of carrot (Daucus carota) is poorly understood. We provide the first such study using a large data set of molecular markers and accessions that are widely dispersed around the world. • METHODS: Sequencing data from the carrot transcriptome were used to develop 4000 single nucleotide polymorphisms (SNPs). Eighty-four genotypes, including a geographically well-distributed subset of wild and cultivated carrots, were genotyped using the KASPar assay. • KEY RESULTS: Analysis of allelic diversity of SNP data revealed no reduction of genetic diversity in cultivated vs. wild accessions. Structure and phylogenetic analysis indicated a clear separation between wild and cultivated accessions as well as between eastern and western cultivated carrot. Among the wild carrots, those from Central Asia were genetically most similar to cultivated accessions. Furthermore, we found that wild carrots from North America were most closely related to European wild accessions. • CONCLUSIONS: Comparing the genetic diversity of wild and cultivated accessions suggested the absence of a genetic bottleneck during carrot domestication. In conjunction with historical documents, our results suggest an origin of domesticated carrot in Central Asia. Wild carrots from North America were likely introduced as weeds with European colonization. These results provide answers to long-debated questions of carrot evolution and domestication and inform germplasm curators and breeders on genetic substructure of carrot genetic resources.

Differential ability of three bee species to move genes via pollen.

Since the release of genetically engineered (GE) crops, there has been increased concern about the introduction of GE genes into non-GE fields of a crop and their spread to feral or wild cross-compatible relatives. More recently, attention has been given to the differential impact of distinct pollinators on gene flow, with the goal of developing isolation distances associated with specific managed pollinators. To examine the differential impact of bee species on gene movement, we quantified the relationship between the probability of getting a GE seed in a pod, and the order in which a flower was visited, or the cumulative distance traveled by a bee in a foraging bout. We refer to these relationships as 'seed curves' and compare these seeds curves among three bee species. The experiments used Medicago sativa L. plants carrying three copies of the glyphosate resistance (GR) allele as pollen donors (M. sativa is a tetraploid), such that each pollen grain carried the GR allele, and conventional plants as pollen recipients. Different foraging metrics, including the number of GR seeds produced over a foraging bout, were also quantified and contrasted among bee species. The lowest number of GR seeds set per foraging bout, and the GR seeds set at the shortest distances, were produced following leafcutting bee visits. In contrast, GR seeds were found at the longest distances following bumble bee visits. Values for honey bees were intermediate. The ranking of bee species based on seed curves correlated well with field-based gene flow estimates. Thus, differential seed curves of bee species, which describe patterns of seed production within foraging bouts, translated into distinct abilities of bee species to move genes at a landscape level. Bee behavior at a local scale (foraging bout) helps predict gene flow and the spread of GE genes at the landscape scale.

Bee species perform distinct foraging behaviors that are best described by different movement models.

In insect-pollinated plants, the foraging behavior of pollinators affects their pattern of movement. If distinct bee species vary in their foraging behaviors, different models may best describe their movement. In this study, we quantified and compared the fine scale movement of three bee species foraging on patches of Medicago sativa. Bee movement was described using distances and directions traveled between consecutive racemes. Bumble bees and honey bees traveled shorter distances after visiting many flowers on a raceme, while the distance traveled by leafcutting bees was independent of flower number. Transition matrices and vectors were calculated for bumble bees and honey bees to reflect their directionality of movement within foraging bouts; leafcutting bees were as likely to move in any direction. Bee species varied in their foraging behaviors, and for each bee species, we tested four movement models that differed in how distances and directions were selected, and identified the model that best explained the movement data. The fine-scale, within-patch movement of bees could not always be explained by a random movement model, and a general model of movement could not be applied to all bee species.

Phylogenetic insights into the correlates of dioecy in meadow-rues (Thalictrum, Ranunculaceae).

Numerous studies have examined the evolution of sexual systems in angiosperms, but few explore the interaction between these and the evolution of pollination mode. Wind pollination is often associated with unisexual flowers, but which evolved first and played a causative role in the evolution of the other is unclear. Thalictrum, meadow-rues (Ranunculaceae), provides a unique opportunity to study the evolution of these traits because it contains insect and wind pollination and four sexual systems. We used a phylogenetic approach to reconstruct ancestral states for sexual system, pollination mode, and geographic distribution in Thalictrum, and tested for correlations to uncover the factors involved in the evolution of unisexuality and wind pollination. Our results show that dioecy, andro- and gynomonoecy evolved at least twice from hermaphroditism. Wind pollination, unisexual flowers, and New World distribution were all significantly correlated. Wind pollination may have evolved early in the genus, followed by multiple losses and gains, and likely preceded the origin of unisexual flowers in several cases; we found no evidence for unisexual flowers evolving prior to wind pollination. Given a broad scale study showing the evolution of dioecy before wind pollination, our results from a finer scale analysis highlight that different evolutionary pathways are likely to occur throughout angiosperms.

Self-Fertilization, Inbreeding, and Yield in Alfalfa Seed Production.

Selfing (self-pollination) is the ultimate form of inbreeding, or mating among close relatives. Selfing can create yield loss when inbreeding depression, defined as a lower survival and reproduction of inbred relative to outbred progeny, is present. To determine the impact of selfing in alfalfa (Medicago sativa L.), we quantified the selfing rate of 32 alfalfa seed production fields located in three regions, namely, the Pacific Northwest (PNW), the Central Valley of California (CEV), and the Imperial Valley of California (IMP). Selfing rates (the proportion of selfed seeds) varied between 5.3 and 30% with an average of 12.2% over the 32 seed production fields. In both the parents and their progeny, we observed an excess of heterozygotes relative to Hardy-Weinberg expectations. We detected notable levels of inbreeding in parents (0.231 ± 0.007 parental inbreeding coefficient) and progeny (0.229 ± 0.005). There were a 15% decrease in the number of seeds per stem (seed set) and a 13% decline in the number of seeds per pod in selfed relative to outcrossed stems, but negligible inbreeding depression for pods per raceme and seed weight. The number of racemes on selfed stems increased significantly in fields with greater selfing rates, supporting the presence of geitonogamous or among flower selfing. Despite the significant level of inbreeding depression, seed set did not decrease in fields with higher selfing rates, where the greater number of racemes on the selfed stems increased the seed set. The effects of the field selfing rate on the seed yield metrics were mostly indirect with direct effects of the number of racemes per stem. Available data indicate that the majority of selfing in alfalfa is pollinator-mediated, and thus, eliminating selfing in alfalfa seed production would require the selection of self-incompatible varieties, which, by eliminating inbreeding depression, would provide a 15% potential increase in seed yield and an increase in future hay yield.

Patch selection by bumble bees navigating discontinuous landscapes.

Pollen and nectar resources are unevenly distributed over space and bees must make routing decisions when navigating patchy resources. Determining the patch selection process used by bees is crucial to understanding bee foraging over discontinuous landscapes. To elucidate this process, we developed four distinct probability models of bee movement where the size and the distance to the patch determined the attractiveness of a patch. A field experiment with a center patch and four peripheral patches of two distinct sizes and distances from the center was set up in two configurations. Empirical transition probabilities from the center to each peripheral patch were obtained at two sites and two years. The best model was identified by comparing observed and predicted transition probabilities, where predicted values were obtained by incorporating the spatial dimensions of the field experiment into each model's mathematical expression. Bumble bees used both patch size and isolation distance when selecting a patch and could assess the total amount of resources available in a patch. Bumble bees prefer large, nearby patches. This information will facilitate the development of a predictive framework to the study of bee movement and of models that predict the movement of genetically engineered pollen in bee-pollinated crops.

Gene flow in commercial alfalfa (Medicago sativa subsp. sativa L.) seed production fields: Distance is the primary but not the sole influence on adventitious presence.

In insect-pollinated crops, gene flow is affected by numerous factors including crop characteristics, mating system, life history, pollinators, and planting management practices. Previous studies have concentrated on the impact of distance between genetically engineered (GE) and conventional fields on adventitious presence (AP) which represents the unwanted presence of a GE gene. Variables other than distance, however, may affect AP. In addition, some AP is often present in the parent seed lots used to establish conventional fields. To identify variables that influence the proportion of AP in conventional alfalfa fields, we performed variable selection regression analyses. Analyses based on a sample-level and a field-level analysis gave similar, though not identical results. For the sample-level model, distance from the GE field explained 66% of the variance in AP, confirming its importance in affecting AP. The area of GE fields within the pollinator foraging range explained an additional 30% of the variation in AP in the model. The density of alfalfa leafcutting bee domiciles influenced AP in both models. To minimize AP in conventional alfalfa seed fields, management practices should focus on optimizing isolation distances while also considering the size of the GE pollen pool within the pollinator foraging range, and the foraging behavior of pollinators.

Phenotypic Selection on Flower Color and Floral Display Size by Three Bee Species.

Plants exhibit a wide array of floral forms and pollinators can act as agent of selection on floral traits. Two trends have emerged from recent reviews of pollinator-mediated selection in plants. First, pollinator-mediated selection on plant-level attractants such as floral display size is stronger than on flower-level attractant such as flower color. Second, when comparing plant species, distinct pollinators can exert different selection patterns on floral traits. In addition, many plant species are visited by a diverse array of pollinators but very few studies have examined selection by distinct pollinators. In the current study, we examined phenotypic selection on flower color and floral display size by three distinct bee species, the European honey bee, Apis mellifera, the common eastern bumble bee, Bombus impatiens, and the alfalfa leafcutting bee, Megachile rotundata, foraging on Medicago sativa. To estimate phenotypic selection by each bee species and for all bees combined simultaneously and on the same group of plants, we introduce a new method that combines pollinator visitation data to seed set and floral trait measurements data typical of phenotypic selection study. When comparing floral traits, all bee species selected on the number of racemes per stem and the number of stems per plant, two components of floral display size. However, only leafcutting bees selected on hue or flower color and only bumble bees selected on chroma or darkness of flowers. Selection on chroma occurred via correlational selection between chroma and number of open flowers per raceme and we examine how correlational selection may facilitate the evolution of flower color in plant populations. When comparing bee species, the three bee species exerted similar selection pattern on some floral traits but different patterns on other floral traits and differences in selection patterns were observed between flower-level and plant-level attractants. The trends detected were consistent with previous studies and we advocate the approach introduced here for future studies examining the impact of distinct pollinators on floral trait evolution.

The influence of distinct pollinators on female and male reproductive success in the Rocky Mountain columbine.

Although there are many reasons to expect distinct pollinator types to differentially affect a plant's reproductive success, few studies have directly examined this question. Here, we contrast the impact of two kinds of pollinators on reproductive success via male and female functions in the Rocky Mountain columbine, Aquilegia coerulea. We set up pollinator exclusion treatments in each of three patches where Aquilegia plants were visited by either day pollinators (majority bumble bees), by evening pollinators (hawkmoths), or by both (control). Day pollinators collected pollen and groomed, whereas evening pollinators collected nectar but did not groom. Maternal parents, potential fathers and progeny arrays were genotyped at five microsatellite loci. We estimated female outcrossing rate and counted seeds to measure female reproductive success and used paternity analysis to determine male reproductive success. Our results document that bumble bees frequently moved pollen among patches of plants and that, unlike hawkmoths, pollen moved by bumble bees sired more outcrossed seeds when it remained within a patch as opposed to moving between patches. Pollinator type differentially affected the outcrossing rate but not seed set, the number of outcrossed seeds or overall male reproductive success. Multiple visits to a plant and more frequent visits by bumble bees could help to explain the lack of impact of pollinator type on overall reproductive success. The increase in selfing rate with hawkmoths likely resulted from the abundant pollen available in experimental flowers. Our findings highlighted a new type of pollinator interactions that can benefit a plant species.

Pollinators of the Rocky Mountain columbine: temporal variation, functional groups and associations with floral traits.

BACKGROUND AND AIMS: Pollinators together with other biotic and some abiotic factors can select for floral traits. However, variation in pollinator abundance over time and space can weaken such selection. In the present study, the variation in pollinator abundance over time and space was examined in populations of the Rocky Mountain columbine. The variation in three floral traits is described and correlations between pollinator type, functional pollinator groups or altitude and floral traits are examined. METHODS: Pollinator observations took place in six Aquilegia coerulea populations over 1-4 years and spur length, flower colour and sepal length were measured in 12 populations. Pollinator abundance, measured as visits per flower per hour, was compared among populations and years. Pollinators were grouped into two functional groups: pollen or nectar collectors. The following associations were examined: annual presence of hawkmoths and whiter flowers with longer spurs; the presence of Sphinx vashti and longer spurs; and higher altitudes and whiter flowers. The study looked at whether an increase in the proportion of hawkmoths in a population was associated with whiter and larger flowers with longer spurs. KEY RESULTS: The abundance of different pollinator groups varied over time and space. Floral traits varied among populations. Higher altitude was correlated with bluer flowers. Whiter flowers were associated with the annual presence of hawkmoths. Populations visited by Sphinx vashti had longer spurs than populations visited only by Hyles lineata. Populations with greater percentage of nectar-collecting pollinators did not have whiter, larger flowers with longer spurs. CONCLUSIONS: Despite the large variation in pollinator abundance over time and space, one species of bumble-bee or hawkmoth tended to predominate in each population each year. Future studies of Aquilegia coerulea should examine the specific influences of pollinators and the environment on flower colour and of hawkmoth species on spur length.

Linking the foraging behavior of three bee species to pollen dispersal and gene flow.

Foraging behaviors that impact gene flow can guide the design of pollinator strategies to mitigate gene flow. Reduced gene flow is expected to minimize the impact of genetically engineered (GE) crops on feral and natural populations and to facilitate the coexistence of different agricultural markets. The goal of this study is to link foraging behavior to gene flow and identify behaviors that can help predict gene flow for different bee species. To reach this goal, we first examined and compared the foraging behaviors of three distinct bee species, the European honey bee, Apis mellifera L., the common eastern bumble bee, Bombus impatiens Cr., and the alfalfa leafcutting bee, Megachile rotundata F., foraging on Medicago sativa flowers. Each foraging behavior investigated differed among bee species. Both social bees exhibited directionality of movement and had similar residence, in contrast to the random movement and shorter residence of the solitary bee. Tripping rate and net distance traveled differed among the three bee species. We ranked each behavior among bee species and used the relative ranking as gene flow predictor before testing the predictions against empirical gene flow data. Tripping rate and net distance traveled, but not residence, predicted relative gene dispersal among bee species. Linking specific behaviors to gene flow provides mechanisms to explain differences in gene flow among bee species and guides the development of management practices to reduce gene flow. Although developed in one system, the approach developed here can be generalized to different plant/pollinator systems.

Impact of insect pollinator group and floral display size on outcrossing rate.

Despite the strong influence of pollination ecology on the evolution of selfing, we have little information on how distinct groups of insect pollinators influence outcrossing rate. However, differences in behavior between pollinator groups could easily influence how each group affects outcrossing rate. We examined the influence of distinct insect pollinator groups on outcrossing rate in the rocky mountain columbine, Aquilegia coerulea. The impact of population size, plant density, size of floral display, and herkogamy (spatial separation between anthers and stigmas) on outcrossing rate was also considered as these variables were previously found to affect outcrossing rate in some plant species. We quantified correlations between all independent variables and used simple and two-factor regressions to determine direct and indirect impact of each independent variable on outcrossing rate. Outcrossing rate increased significantly with hawkmoth abundance but not with the abundance of any of the other groups of floral visitors, which included bumblebees, solitary bees, syrphid flies, and muscidae. Outcrossing rate was also significantly affected by floral display size and together, hawkmoth abundance and floral display size explained 87% of the variation in outcrossing rate. None of the other independent variables directly affected the outcrossing rate. This is the first report of a significant impact of pollinator type on outcrossing rate. Hawkmoths did not visit fewer flowers per plant relative to other pollinator groups but preferred visiting female-phase flowers first on a plant. Both the behavior of pollinators and floral display size affected outcrossing rate via their impact on the level of geitonogamous (among flower) selfing. Given that geitonogamous selfing is never advantageous, the variation in outcrossing rate and maintenance of mixed mating systems in populations of A. coerulea may not require an adaptive explanation.

The extent of hybridization and its impact on the genetic diversity and population structure of an invasive tree, Ulmus pumila (Ulmaceae).

Ulmus pumila is considered an invasive tree in 41 of the United States. In this study, we examined the extent of hybridization in naturalized populations of U. pumila, its impact on genetic diversity and genetic structure and its potential role in explaining the invasion process of U. pumila. Genetic analyses indicated widespread hybridization with native Ulmus rubra in naturalized U. pumila populations. Hybridization increased the genetic diversity of U. pumila populations and affected their genetic structure. The level of genetic diversity in 'mature' accessions, many of which may represent original plantings throughout the USA, was high and similar to the diversity of East Asian accessions. Hybridization with the native red elm may play an important role in the success of Siberian elm as an invader in temperate regions of the USA.

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.