Genetic structure in aquatic bladderworts: clonal propagation and hybrid perpetuation

• Background and Aims The free-floating aquatic bladderwort Utricularia australis f. australis is a sterile F₁ hybrid of U. australis f. tenuicaulis and U. macrorhiza. However, co-existence of the hybrids and parental species has not been observed. In the present study, the following questions are addressed. (a) Does the capacity of the two parental species to reproduce sexually contribute to higher genotypic diversity than that of sterile F₁ hybrid? (b) Are there any populations where two parental species and their hybrid co-exist? (c) If not, where and how do hybrids originate?• Methods The presence and absence of Utricularia was thoroughly investigated in two regions in Japan. An amplified fragment length polymorphism (AFLP) analysis was conducted for 397 individuals collected from all populations (33 in total) where Utricularia was observed.• Key Results The mean number of genotypes per population (G) and genotypic diversity (D) were extremely low irrespective of the capacity to reproduce sexually: G was 1·1–1·2 and D was 0·02–0·04. The hybrid rarely co-existed with either parental species, and the co-existence of two parental species was not observed. Several AFLP bands observed in the hybrid are absent in both parental genotypes, and parent and hybrid genotypes in the same region do not show greater genetic similarity than those in distant regions.• Conclusions The capacity to reproduce sexually in parental species plays no role in increasing genotypic diversity within populations. The observed genotypes of the hybrid could not have originated from hybridization between the extant parental genotypes within the study regions. Considering the distribution ranges of three investigated taxa, it is clear that the hybrid originated in the past, and hybrid populations have been maintained exclusively by clonal propagation, which may be ensured by both hybrid vigor and long-distance dispersal of clonal offspring.     

Variation of sterility and fertility in Utricularia australis f. australis in Hokkaido, northern Japan

Seed-production ability was studied in Utricularia australis R. Br. f. australis Komiya and Shibata through field observation, pollination experiment, pollen culture, and isozyme analysis for populations in various regions of Hokkaido, northern Japan. Utricularia australis f. australis has previously been postulated to be sterile in Japan, however, in the present study, fertile populations were found in eastern Hokkaido. Bisexual sterility and male sterility were suspected to be present in populations in the western part of Hokkaido. There was also a strain that seemed to produce no seed because of intra-clonal inbreeding depression or self-incompatibility. Isozyme analysis of three enzyme systems indicated a uniformity of multienzymatic phenotype (MEP) in each population or region. The MEP grouping of populations corresponded to the type of sterility and fertility revealed by pollination and pollen culture experiments. These experiments suggested that each regional strain had its own sterility or fertility type.     

Microbial colonization and competition on the marine alga Ulva australis

Pseudalteromonas tunicata and Roseobacter gallaeciensis are biofilm-forming marine bacteria that are often found in association with the surface of the green alga Ulva australis. They are thought to benefit the plant host by producing inhibitory compounds that are active against common fouling organisms. We investigated factors that influence the ability of P. tunicata and R. gallaeciensis to attach to and colonize the plant surface and also the competitive interactions that occur between these organisms and other isolates from U. australis during biofilm formation on the plant surface. A surprisingly high number of P. tunicata cells, at least 10(8) cells ml(-1), were required for colonization and establishment of a population of cells that persists on axenic surfaces of U. australis. Factors that enhanced colonization of P. tunicata included inoculation in the dark and pregrowth of inocula in medium containing cellobiose as the sole carbon source (cellulose is a major surface polymer of U. australis). It was also found that P. tunicata requires the presence of a mixed microbial community to colonize effectively. In contrast, R. gallaeciensis effectively colonized the plant surface under all conditions tested. Studies of competitive interactions on the plant surface revealed that P. tunicata was numerically dominant compared with all other bacterial isolates tested (except R. gallaeciensis), and this dominance was linked to production of the antibacterial protein AlpP. Generally, P. tunicata was able to coexist with competing strains, and each strain existed as microcolonies in spatially segregated regions of the plant. R. gallaeciensis was numerically dominant compared with all strains tested and was able to invade and disperse preestablished biofilms. This study highlighted the fact that microbial colonization of U. australis surfaces is a dynamic process and demonstrated the differences in colonization strategies exhibited by the epiphytic bacteria P. tunicata and R. gallaeciensis.     

Clonal genetic diversity and populational genetic differentiation in Phragmites australis distributed in the Songnen Prairie in northeast China as revealed by amplified fragment length polymorphism and sequence-specific amplification polymorphism molecular markers

Genetic variation within and between four naturally occurring Phragmites australis land populations, DBS, QG, SS1 and SS2 (named after locality), which colonise distinct habitats (different edaphic conditions) in the Songnen Prairie in northeast China, were investigated by amplified fragment length polymorphism (AFLP) and sequence‐specific amplification polymorphism (S‐SAP) markers. It was found that the selected primer combinations of both markers were highly efficient in revealing the inter‐clonal genetic diversity and inter‐populational genetic differentiation in P. australis from a molecular ecological perspective. Cluster analysis categorised the plants into distinct groups (DBS, QG and SS groups), which were in line with their localities, albeit the two SS group populations (SS1 and SS2) showed a lower degree of inter‐populational differentiation. These results were strongly supported by multiple statistical analysis including Mantel’s test, principal coordinate analysis, allocation test and analysis of molecular variance, which further suggested that gene flow, genetic drift and differences in as yet unidentified edaphic factors may all underpin the inter‐clonal genetic diversity and inter‐populational differentiation at the nucleotide sequence level. Analysis of intra‐population clonal diversity also revealed that the QG population harboured a strikingly lower amount of within‐population variation compared with those of the other three populations, presumably being caused by genetic drift and followed by physical and/or biological isolation. Homology analysis of a subset of population‐specific or population‐private AFLP and S‐SAP bands suggested that regulatory genes and retroelements might play important roles in the ecological adaptation and differentiation of the P. australis populations. Possible causes for and implications of the extensive genetic variability in P. australis were discussed for its future genetic conservation and use in ecological revegetation.     

Influence of die-back syndrome on reproductive strategies within Phragmites australis populations

Sexual plant reproduction is a strategy that allows plant populations to increase genetic variability, and consequently to be more efficient in adapting to new environments and to overcome stress conditions. Here, we focus on the reproductive mode of Phragmites australis, an important sub-cosmopolite species that can spread both by clonal propagation and sexual reproduction. In Europe, P. australis is affected by severe decline (known as Reed Die-Back Syndrome or RDBS); this study aims to understand if in RDBS condition the reproductive strategy of P. australis was altered, based on the hypothesis that if stress occurs, plants are likely to use their energy to increase genetic variability to overcome the stress. Inflorescences in five Italian wetlands were sampled from healthy and RDBS-affected stands of P. australis. Seed production, seed viability and seed germination were evaluated, with germination monitored under two different water conditions: moist and submerged. Stress induced by RDBS appears to influence the reproductive strategy of P. australis. In RDBS-affected stands, seed production was significantly higher than healthy stands for four of five sites. Seed viability and germination were also higher in RDBS-affected stands, but these differences were lower and site dependent.     

扎龙芦苇遗传多样性ISSR分析

采用ISSR分子标记技术对扎龙湿地的5个野生芦苇居群进行了遗传多样性研究.从75条ISSR引物中筛选出10个特异性强、稳定性好的引物进行ISSR分析.共获得96个位点,其中多态位点94个,多态位点百分率为97.92％.PopGene分析结果表明:居群的平均多态位点百分率为56.46％,Shannon's多样性指数(J)...     

Evidence for a switch in the reproductive biology of Rubus alceifolius (Rosaceae) towards apomixis, between its native range and its area of introduction

We compared the reproductive system of Rubus alceifolius in its native range in Southeast Asia, in Madagascar, where the plant was introduced apparently some centuries ago, and in La Réunion, an Indian Ocean island onto which R. alceifolius was introduced (from Madagascan source populations) around 1850. While tetraploidy makes it impossible to analyze variation in R. alceifolius using classical methods of population genetics, both the patterns of genetic diversity (as revealed by AFLP [amplified fragment length polymorphism] markers) and differences between half-sib progeny and their maternal parents (revealed by microsatellite markers) show that in the plant's native range in southeast Asia, seeds are produced sexually. In contrast, in Madagascar sexual reproduction cannot alone account for the genetic patterns observed with microsatellite markers. Over 85% of the half-sib progeny resulting from open pollination gave multilocus genotypes identical to those of their respective maternal parents, despite the fact that the latter had alleles that were rare in the population. The other progeny differed in having an allele with one motif more or less than that of the maternal parent. Seeds thus appear to be produced mostly or exclusively by apomixis in Madagascar. We present findings suggesting that Madagascan populations result from hybridization of introduced R. alceifolius and native populations of R. roridus, a closely related species of Rubus subgenus Malachobatus, and suggest that apomixis was a consequence of this hybridization. In Reunionese populations of R. alceifolius (derived from Madagascan populations), seeds obtained in controlled pollination experiments were all genetically identical to maternal parents. While genetic variation (microsatellite markers) in Reunionese populations was low, it was sufficient to allow us to demonstrate that seeds could not have resulted from fertilization by the pollen donors chosen for controlled pollinations, or from autogamy, and were produced exclusively by apomixis.     

Genetic characterization of hybridization and introgression between anadromous rainbow trout (Oncorhynchus mykiss irideus) and coastal cutthroat trout (O. clarki clarki)

Interspecific hybridization represents a dynamic evolutionary phenomenon and major conservation problem in salmonid fishes. In this study we used amplified fragment length polymorphisms (AFLP) and mitochondrial DNA (mtDNA) markers to describe the extent and characterize the pattern of hybridization and introgression between coastal rainbow trout (Oncorhynchus mykiss irideus) and coastal cutthroat trout (O. clarki clarki). Hybrid individuals were initially identified using principle coordinate analysis of 133 polymorphic AFLP markers. Subsequent analysis using 23 diagnostic AFLP markers revealed the presence of F1, rainbow trout backcross, cutthroat trout backcross and later-generation hybrids. mtDNA analysis demonstrated equal numbers of F1 hybrids with rainbow and cutthroat trout mtDNA indicating reciprocal mating of the parental types. In contrast, rainbow and cutthroat trout backcross hybrids always exhibited the mtDNA from the recurrent parent, indicating a male hybrid mating with a pure female. This study illustrates the usefulness of the AFLP technique for generating large numbers of species diagnostic markers. The pattern of hybridization raises many questions concerning the existence and action of reproductive isolating mechanisms between these two species. Our findings are consistent with the hypothesis that introgression between anadromous populations of coastal rainbow and coastal cutthroat trout is limited by an environment-dependent reduction in hybrid fitness.     

Multifaceted,Cross-Generational Costs of Hybridization in Sibling Drosophila species

Maladaptive hybridization, as determined by the pattern and intensity of selection against hybrid individuals, is an important factor contributing to the evolution of prezygotic reproductive isolation. To identify the consequences of hybridization between Drosophila pseudoobscura and D. persimilis, we estimated multiple fitness components for F1 hybrids and backcross progeny and used these to compare the relative fitness of parental species and their hybrids across two generations. We document many sources of intrinsic (developmental) and extrinsic (ecological) selection that dramatically increase the fitness costs of hybridization beyond the well-documented F1 male sterility in this model system. Our results indicate that the cost of hybridization accrues over multiple generations and reinforcement in this system is driven by selection against hybridization above and beyond the cost of hybrid male sterility; we estimate a fitness loss of >95% relative to the parental species across two generations of hybridization. Our findings demonstrate the importance of estimating hybridization costs using multiple fitness measures from multiple generations in an ecologically relevant context; so doing can reveal intense postzygotic selection against hybridization and thus, an enhanced role for reinforcement in the evolution of populations and diversification of species.     

The status of Clarkia australis (Onagraceae)

Clarkia australis and C. virgata grow on the western slope of the central Sierra Nevada of California. Clarkia australis was established to accommodate populations of C. virgata from south of the Tuolumne River that could not be successfully hybridized to populations north of the river. Although the species is maintained in the new Jepson Manual, its validity has been questioned because only two populations were originally tested, and they had no useful morphological traits that distinguished them from C. virgata. We report here the results of a large program of interpopulation hybridizations that show that C. australis is distinct and that its reproductive isolation from C. virgata is complete and absolute and reflects a compatibility block that apparently causes abortion of hybrid seeds in early development. Both species include populations north and south of the Tuolumne River and, in general, those of C. australis occupy higher elevations. Morphologically, the species are extremely similar though the mean values of several dimensions of the petals are different. However, significant variation among their populations has the consequence that, at present, the only certain way to assign particular populations to species is to test their compatibility with previously tested populations.     

Dispersal of Udonella australis (Monogenea: Udonellidae) between caligid copepods Caligus rogercresseyi and Lepeophtheirus mugiloidis on Chilean rock cod

Udonella australis is a platyhelminth that lives on the surface of the ectoparasite copepods Caligus rogercresseyi and Lepeophtheirus mugiloidis, which coexist on the Chilean rock cod Eleginops maclovinus. The absence of a planktonic oncomiracidium stage in the life cycle of udonellids may limit their dispersal ability. However, the high prevalence and intensity of U. australis on C. rogercresseyi suggest they have developed dispersal strategies to compensate for the lack of a free-living larval stage. The goals of this study were to determine the main dispersal mechanisms of U. australis in 1 copepod species and to compare the dispersal ability of U. australis between 2 different copepod species. Chilean rock cods were infected with female (without udonellids) and male (with and without udonellids) C. rogercresseyi. Other fishes were also infected with this copepod (with U. australis) and with L. mugiloidis (without U. australis). The dispersal of udonellids among copepods occurs through both intraspecific and interspecific processes. The main dispersal mechanism appears to be copepod mating; contact between same-sex individuals is less important. Intraspecific dispersal seems to be more dependent on the number of udonellids per fish than on copepod abundance, as observed for interspecific dispersal.     

LOCAL ADAPTATION,PHENOTYPIC DIFFERENTIATION,AND HYBRID FITNESS IN DIVERGED NATURAL POPULATIONS OF ARABIDOPSIS LYRATA

Selection for local adaptation results in genetic differentiation in ecologically important traits. In a perennial, outcrossing model plant Arabidopsis lyrata, several differentiated phenotypic traits contribute to local adaptation, as demonstrated by fitness advantage of the local population at each site in reciprocal transplant experiments. Here we compared fitness components, hierarchical total fitness and differentiation in putatively ecologically important traits of plants from two diverged parental populations from different continents in the native climate conditions of the populations in Norway and in North Carolina (NC, U.S.A.). Survival and number of fruits per inflorescence indicated local advantage at both sites and aster life‐history models provided additional evidence for local adaptation also at the level of hierarchical total fitness. Populations were also differentiated in flowering start date and floral display. We also included reciprocal experimental F₁ and F₂ hybrids to examine the genetic basis of adaptation. Surprisingly, the F₂ hybrids showed heterosis at the study site in Norway, likely because of a combination of beneficial dominance effects from different traits. At the NC site, hybrid fitness was mostly intermediate relative to the parental populations. Local cytoplasmic origin was associated with higher fitness, indicating that cytoplasmic genomes also may contribute to the evolution of local adaptation.     

Natural hybridization in Cardamine (Brassicaceae) in the Pyrenees: evidence from morphological and molecular data

While researching Cardamine (Brassicaceae) in the Pyrenees, putative hybrid plants were found at two natural sites. Pollen grain viability, AFLP, and multivariate morphometric analyses were performed in order to assess the plants' presumed hybrid origin, establishing that natural hybridization between the diploids C. crassifolia and C. amara ssp. pyrenaea had occurred. A new diploid nothospecies, C. × enriquei (2 n = 2x = 16), is described. Examination of 18 morphological characters showed the intermediacy of the hybrid between the parental taxa in most characters. AFLP analyses of C. amara ssp. amara , ssp. austriaca , ssp. olotensis and C. raphanifolia , demonstrated the close position of the hybrid to C. crassifolia , and revealed that the highest number of markers were shared with the parents. Polymorphism found in the AFLP pattern of the hybrid suggested recurrent origin, segregation and/or backcrosses, although assessment of pollen viability indicated high male sterility. The hybridization event reported here represents the second documented case between the C. pratensis group and C. amara . An account of the nomenclature of C. crassifolia is also presented, including lectotypification of relevant names.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 275–294.     

敦煌阳关盐沼湿地芦苇种群小尺度空间分布格局

植物种群的空间分布格局是研究连接植物与外界环境关系的重要桥梁,是植物种群稳定性和生活史策略的表现形式。本研究以敦煌阳关盐沼湿地克隆植物芦苇(Phragmites australis)为对象,设置湿生区、轻度盐化区、重度盐化区和荒漠区采样梯度,应用Ripley K(d)函数分析法研究了芦苇种群的小尺度空间分布格局特征及生态适应对策。结果表明:(1)随着从湿生到荒漠区环境梯度的变化,芦苇种群的盖度、密度、高度、地上生物量及种群领地面积都呈现出显著下降的趋势,空斑面积呈现出显著增加的趋势,而种群领地密度则呈现出先增加后下降的趋势;(2)从湿生到荒漠生境,芦苇种群的小尺度空间拓展策略主要表现为由聚集分布为主、非聚集分布(随机分布或均匀分布)为辅,转变为随机分布为主、非随机分布(聚集分布或均匀分布)为辅的特征。研究结果明晰了干旱区内陆河湿地中芦苇种群的空间分布格局特点及其影响因素,为进一步理解克隆植物的生态适应机制提供了案例研究,为科学保护和管理干旱区内陆河湿地生态系统提供了理论依据。     

Population genetic structure of <Emphasis Type="Italic">Pyrenophora teres</Emphasis>Drechs. the causal agent of net blotch in Sardinian landraces of barley (<Emphasis Type="Italic">Hordeum vulgare L.</Emphasis>)

Monoconidial cultures of Pyrenophora teres, the causal agent of barley net blotch, were isolated from leaves collected from six populations of the barley landrace "S'orgiu sardu" growing in five agro-ecological areas of Sardinia, Italy, and genotyped using AFLPs. The 150 isolates were from lesions of either the "net form" (P. teres f. sp. teres) or the "spot form" (P. teres f. sp. maculata) of the disease. Of 121 AFLP markers, 42%, were polymorphic. Cluster analysis resolved the isolates into two strongly divergent groups (F(ST) = 0.79), corresponding to the net (45% of the isolates) and the spot (55% of the isolates) forms (designated the NFR and SFR groups, respectively). The absence of intermediate genotypes and the low number of shared markers between the two groups indicated that hybridization between the two formae is rare or absent under the field condition of Sardinia. Five of the barley populations hosted both forms but in different proportions. The SFR populations were similar in overall polymorphism to the NFR populations. However, compared to the SFR form, the NFR occurred in all fields sampled and showed a higher population divergence (F(ST) = 0.43 versus F(ST) = 0.09 with all isolates; F(ST) = 0.37 versus F(ST) = 0.06 with clone corrected samples) probably due to a lower migration rate. AFLP fingerprints resolved 117 distinct genotypes among the 150 isolates sampled (78%), 87% in SFR and 68% in NFR isolates. Although the absolute numbers may be a function of the number of AFLP markers assayed, the relative difference suggests that clonality is more prevalent among the NFR isolates (with 11 of 46 haplotypes observed more than once), compared with SFR isolates (7 of 71 haplotypes). Both digenic and multilocus linkage disequilibrium analyses suggested that sexual reproduction occurs at significant levels within the NFR and SFR populations, and that the relative contribution of sexual and asexual reproduction varies among different environments.     

Population genetic structure in European populations of Spiranthes romanzoffiana set in the context of other genetic studies on orchids

Spiranthes romanzoffiana Cham. is restricted in Europe to the British Isles, where it is recognised as a conservation priority species due to frequent extirpation of populations along with no evidence of seed set; vegetative reproduction has been invoked as the sole means of perpetuation and dispersal. To investigate the reproductive ecology of this species, 17 populations have been sampled for chloroplast microsatellites and amplified fragment length polymorphisms (AFLPs). These markers revealed a previously unsuspected genetic-geographic split in the species, which correlates with differences in patterns of within-population variation. Northern populations were fixed for one chloroplast haplotype but showed high levels of AFLP genotypic diversity consistent with sexual reproduction (proportion of genotypes distinguishable, PD = 0.98). More southerly populations showed fixed differences from the northern populations in their chloroplast haplotype and for 10 AFLP markers. They harboured only 12 unique multilocus genotypes among 113 individuals from six populations (PD = 0.11). These genotypes differed mostly by single bands, and none by more than 4/138 loci, with identical multilocus genotypes occurring in widely separated populations. This uniformity in southern populations is consistent with agamospermous or autogamous reproduction, and/or an extreme population bottleneck. Finally, the observed patterns of population differentiation in S. romanzoffiana are compared with other studies of orchids, revealing a wide range of values that belie recent contrasting published generalisations that claim that orchids show either higher, or lower, levels of population differentiation than other plant families.     

Isolation and characterization of the mating-type locus of the barley pathogen Pyrenophora teres and frequencies of mating-type idiomorphs within and among fungal populations collected from barley landraces.

Pyrenophora teres f. sp. teres mating-type genes (MAT-1: 1190 bp; MAT-2: 1055 bp) have been identified. Their predicted proteins, measuring 379 and 333 amino acids, respectively, are similar to those of other Pleosporales, such as Pleospora sp., Cochliobolus sp., Alternaria alternata, Leptosphaeria maculans, and Phaeosphaeria nodorum. The structure of the MAT locus is discussed in comparison with those of other fungi. A mating-type PCR assay has also been developed; with this assay we have analyzed 150 isolates that were collected from 6 Sardinian barley landrace populations. Of these, 68 were P. teres f. sp. teres (net form; NF) and 82 were P. teres f. sp. maculata (spot form; SF). Within each mating type, the NF and SF amplification products were of the same length and were highly similar in sequence. The 2 mating types were present in both the NF and the SF populations at the field level, indicating that they have all maintained the potential for sexual reproduction. Despite the 2 forms being sympatric in 5 fields, no intermediate isolates were detected with amplified fragment length polymorphism (AFLP) analysis. These results suggest that the 2 forms are genetically isolated under the field conditions. In all of the samples of P. teres, the ratio of the 2 mating types was consistently in accord with the 1:1 null hypothesis. This ratio is expected when segregation distortion and clonal selection among mating types are absent or asexual reproduction is rare. Overall, sexual reproduction appears to be the major process that equalizes the frequencies of the 2 mating types within populations.     

Hybrid fitness in a locally adapted parasite

The parasite (Red Queen) hypothesis for the maintenance of sexual reproduction and genetic diversity assumes that host-parasite interactions result from tight genetic specificity. Hence, hybridization between divergent parasite populations would be expected to disrupt adaptive gene combinations, leading to reduced infectivity on exposure to parental sympatric hosts, as long as gene effects are nonadditive. In contrast, hybridization would not cause reduced infectivity on allopatric hosts unless the divergent parasite populations possess alleles that are intrinsically incompatible when they are combined. In three different experiments, we compared the infectivity of locally adapted parasite (trematode) populations with that of F(1) hybrid parasites when exposed to host (snail) populations that were sympatric to one of the two parasite populations. We tested for intrinsic genetic incompatibilities in two experiments by including one host population that was allopatric to both parasite populations. As predicted, when the target host populations were sympatric to the parasite populations, the hybrids were significantly less infective than the parental average, while hybrid parasites on allopatric hosts were not, thereby ruling out intrinsic genetic incompatibilities. The results are consistent with nonadditive gene effects and tightly specific host-driven selection underlying parasite divergence, as envisioned by coevolutionary theory and the Red Queen hypothesis.     

Reproductive behavior in the squid Sepioteuthis australis from South Australia: interactions on the spawning grounds

Squid behavior is synonymous with distinctive body patterns, postures, and movements that constitute a complex visual communication system. These communications are particularly obvious during reproduction. They are important for sexual selection and have been identified as a potential means of species differentiation. Here we present a detailed account of copulation, mating, and egg deposition behaviors from in situ observations of the squid Sepioteuthis australis from South Australia. We identified four mating types from 85 separate mating attempts: "Male-upturned mating" (64% of mating attempts); "Sneaker mating" (33%); "Male-parallel" (2%); and "Head-to-head" (1%). Intervals between successive egg deposition behaviors were clearly bimodal, with modes at 2.5 s and 70.0 s. Ninety-three percent of egg capsules contained 3 or 4 eggs (mean = 3.54), and each egg cluster contained between 218 and 1922 egg capsules (mean = 893.9). The reproductive behavior of S. australis from South Australia was different from that described for other cephalopod species. More importantly, comparison between these results and those for other populations of S. australis suggests that behavior may differ from one population to another.     

Asymmetric gene introgression in two closely related Orchis species: evidence from morphometric and genetic analyses

ABSTRACT: BACKGROUND: In food-deceptive orchids of the genera Anacamptis, Neotinea and Orchis floral isolation has been shown to be weak, whereas late-acting reproductive barriers are mostly strong, often restricting hybridization to the F1 generation. Only in a few species hybridization extends beyond the F1 generation, giving rise to hybrid swarms. However, little is known about the abundance of later-generation hybrids and what factors drive their occurrence in hybrid populations. In this study, molecular analyses were combined with detailed morphological measurements in a hybrid population of two closely related Orchis species (Orchis militaris and O. purpurea) to investigate the hypothesis that the abundance of later-generation hybrids is driven by changes in floral characters after hybridization that exert selective pressures that in turn affect hybridization. RESULTS: Both the molecular and morphological data point to extensive genetic and morphological homogenization and asymmetric introgression. Estimating genomic clines from the multi-locus genotype data and testing for deviation from neutrality revealed that 30 out of 113 (27%) AFLP markers significantly deviated from neutral expectations. Plants with large floral displays or plant with flowers that resembled more O. purpurea had higher female fitness than plants with small floral displays or plants with flowers resembling more O. militaris, suggesting that directional selection may have contributed to the observed patterns of introgression. CONCLUSIONS: These results indicate that in closely related orchid species hybridization and gene introgression may be partly driven by selection for floral traits of one of the parental types. However, because some pure individuals were still present in the studied population, the parental species appeared to be sufficiently isolated to survive the challenge of sympatry.