Pollen competition as an asymmetric reproductive barrier between two closely related Silene species

Reproductive barriers are important determinants of gene flow between divergent populations or species. We studied pollen competition as a post‐mating reproductive barrier between Silene dioica and S. latifolia. Gene flow between these species is extensive, but early‐generation hybrids are rare. In an experiment with conspecific, heterospecific and 50 : 50 mixed pollinations in the two species, pollination treatments did not significantly affect seed set and seed weight. However, molecular determination of siring success after mixed pollinations showed that fewer than expected hybrids were produced in S. latifolia (18% hybrids) but not in S. dioica (51% hybrids). This constitutes an asymmetric post‐mating reproductive barrier and likely contributes to the rarity of early‐generation hybrids. Our study shows that pollen competition can be an effective barrier to hybridization between closely related species that likely acts in concert with other reproductive barriers.     

Pollen flow and post-pollination barriers in two varieties of Dactylorhiza incarnata s.l. (Orchidaceae)

Dactylorhiza orchids are known for their high variation in morphology and distinct varieties have been named in D. incarnata s.l. However, it is not known how these varieties interact in mixed populations and why they remain stable. We conducted three field experiments in West-Estonian populations of D. incarnata to examine if the two most common varieties co-occurring these are separated from each other either by pre-pollination or post-pollination reproductive barriers. We found that pollinia were far more frequently transferred between the purple-flowered var. incarnata and the pale-flowered var. ochroleuca than between plants of the same variety. Furthermore, in hand-pollination and germination experiments concerning the same two varieties, we found that pollen source (self-pollination, within- or between-varieties pollination) did not affect seed production or probability of fungus infection of the germinating seeds. These two varieties of D. incarnata thus had no pre-pollination or early functioning post-pollination reproductive barriers. Post-pollination barriers may, however, act later in seedling or adult stage.     

Capturing Genetic Variation during Ecological Restorations: An Example from Kankakee Sands in Indiana

Genetic variation in populations, both natural and restored, is usually considered crucial for response to short‐term environmental stresses and for long‐term evolutionary change. To have the best chance of successful long‐term survival, restored populations should reflect the extant variation found in remnants, but restored sites may suffer from genetic bottlenecks as a result of founder effects. Kankakee Sands is a large‐scale restoration being conducted by The Nature Conservancy (TNC) in northwestern Indiana. Our goal was to test for loss of genetic variation in restored plant populations by comparing them with TNC’s seed source nursery and with local remnant populations that were the source of nursery seed and of the first few restored sites. Allozyme analysis of Baptisia leucantha, Asclepias incarnata, Coreopsis tripteris, and Zizia aurea showed low levels of allozyme diversity within all species and reductions in polymorphism, alleles per locus, and expected heterozygosity between remnants and restorations for all species except A. incarnata. Almost all lost alleles were rare; restored populations contained almost 90% of alleles at polymorphic loci that occurred in remnants at frequencies greater than 1%. Allele frequencies for most loci did not differ between remnants and restored sites. Most species showed significant allele frequency differentiation among remnant populations and among restored sites. Our results indicate that seed collection techniques used at Kankakee Sands captured the great majority of allozyme variation present in seed source remnant populations.     

Pollinator-mediated mating restriction between sympatric varieties of yellow lady’s slipper orchids (Cypripedium parviflorum Salisb.)

A predominant theme in the study of orchid evolution has been the importance of floral traits contributing to pollinator-mediated isolating barriers (i.e., floral isolation). However, few studies have quantified the contribution of floral isolation in sympatric orchid populations. Cypripedium parviflorum vars. makasin and pubescens are excellent taxa to test the strength of floral isolation because of their recent phylogenetic separation, overlapping flowering phenologies, and sympatric populations that can lack intermediate morphologies. In this paper, we use sympatric populations to (1) test for pre-mating and early-acting post-mating reproductive isolating barriers, (2) quantify genetic differentiation among populations using allozyme loci, and (3) document floral size differences between varieties. Pollen tracking experiments using fluorescent powders revealed minimal inter-varietal pollen movement. Across two sympatric sites, only 4 inter-varietal pollinations were observed among 52 var. makasin pollinations and 40 var. pubescens pollinations. All of these inter-varietal pollinations had var. makasin as the pollen donor. In contrast, artificial crosses within and between the varieties revealed no statistically significant differences in fruit set, seed weight, or embryo morphology. Allozyme data are consistent with restricted inter-varietal gene flow, showing strong allele frequency differences between the varieties across multiple loci. In a clustering analysis, these differences caused the varieties to group taxonomically and not by their geographic location. These data along with statistically significant morphological differences between the varieties strongly suggest the existence of a pollinator-mediated isolating barrier. The possible nature of the isolating barrier is discussed as well as the likelihood of other late acting post-zygotic barriers.     

Better common than rare? Effects of low reproductive success,scarce pollinator visits and interspecific gene flow in threatened and common species of Tibouchina (Melastomataceae)

The reasons for plant rarity have been the focus of many studies, especially because rare species are more prone to extinction than common species. Under the same habitat conditions, rare plants are expected to attract fewer flower visitors and to show some limitation in their reproductive success. Here, using one of the most emblematic Neotropical plant genus (Tibouchina) we tested whether narrow endemic and threatened species in Ecuador have a lower reproductive success or are visited by fewer pollinators than common species, in 13 populations monitored from 2011 to 2013. We also assessed whether interspecific gene flow could be considered a threat to the rare species. However, contrary to expectations, we found that few pollinators visited the flowers, independently of species rarity. Natural outcross pollinations were always very low in all small‐size populations, leading to high levels of pollen limitation. Interspecific crossing experiments also revealed weak reproductive barriers in some species. This study reveals that both narrow and common species of Tibouchina have similar reproductive and pollinator patterns in Ecuador and, therefore, other causes of the rarity of these species should be considered.     

Oviposition preference and larval performance of North American monarch butterflies on four Asclepias species

Monarch butterflies, Danaus plexippus L. (Lepidoptera: Nymphalidae), occur world‐wide and are specialist herbivores of plants in the milkweed family (Asclepiadaceae). In North America, two monarch populations breed east and west of the continental divide in areas populated by different host plant species. To examine the population variation in monarch responses to different Asclepias species, we measured oviposition preference and larval performance among captive progeny reared from adult butterflies collected in eastern and western North America. Host plant use was evaluated using two milkweed species widely distributed in eastern North America (A. incarnata and A. syriaca), and two species common to western North America (A. fascicularis and A. speciosa). We predicted that exposure to different host plant species in their respective breeding ranges could select for divergent host use traits, so that monarchs should preferentially lay more eggs on, and larvae should perform better on, milkweed species common to their native habitats. Results showed that across all adult female butterflies, oviposition preferences were highest for A. incarnata and lowest for A. fascicularis, but mean preferences did not differ significantly between eastern and western monarch populations. Larvae from both populations experienced the highest survival and growth rates on A. incarnata and A. fascicularis, and we again found no significant interactions between monarch source population and milkweed species. Moreover, the average rank order of larval performance did not correspond directly to mean female oviposition preferences, suggesting that additional factors beyond larval performance influence monarch oviposition behavior. Finally, significant family level variation was observed for both preference and performance responses within populations, suggesting an underlying genetic variation or maternal effects governing these traits.     

Genetic diversity and reproductive ecology of the sage-leaved rockrose,Cistus salviifolius L., in the Swiss Alps

Range marginal populations are often perceived to have lower conservation value compared to those in the core area. The allocation of resources to maintain peripheral populations is therefore often questioned. The sage-leaved rockrose (Cistus salviifolius L.) is a self-incompatible and obligate seeder widely distributed in the Mediterranean area but rare and patchily distributed in Switzerland at its range margin on the southern slopes of the Alps. Here, we combined analysis of genetic diversity with pollinator surveys and field studies of reproductive ecology to compare peripheral Cistus populations in the Alps with range central populations in the Mediterranean. Our results showed no differences in genetic diversity between peripheral and central populations and between fragmented and connected ones at its range margin in the Alps. Although the fragmented populations were visited by more abundant and species richer pollinators (bees and wasps), they showed lower number of seeds and higher self-compatibility compared to the connected ones, which excludes the pollination limitation hypothesis. Overall, our study highlights that peripheral populations of C. salviifolius in the Alps are likely to contribute to maintain genetic diversity, while showing variation in reproductive ecology, and are therefore important for the conservation of this species.

     

Components of reproductive isolation between the monkeyflowers Mimulus lewisii and M. cardinalis (Phrymaceae)

Evolutionists have long recognized the role of reproductive isolation in speciation, but the relative contributions of different reproductive barriers are poorly understood. We examined the nature of isolation between Mimulus lewisii and M. cardinalis, sister species of monkeyflowers. Studied reproductive barriers include: ecogeographic isolation; pollinator isolation (pollinator fidelity in a natural mixed population); pollen competition (seed set and hybrid production from experimental interspecific, intraspecific, and mixed pollinations in the greenhouse); and relative hybrid fitness (germination, survivorship, percent flowering, biomass, pollen viability, and seed mass in the greenhouse). Additionally, the rate of hybridization in nature was estimated from seed collections in a sympatric population. We found substantial reproductive barriers at multiple stages in the life history of M. lewisii and M. cardinalis. Using range maps constructed from herbarium collections, we estimated that the different ecogeographic distributions of the species result in 58.7% reproductive isolation. Mimulus lewisii and M. cardinalis are visited by different pollinators, and in a region of sympatry 97.6% of pollinator foraging bouts were specific to one species or the other. In the greenhouse, interspecific pollinations generated nearly 50% fewer seeds than intraspecific controls. Mixed pollinations of M. cardinalis flowers yielded >75% parentals even when only one-quarter of the pollen treatment consisted of M. cardinalis pollen. In contrast, both species had similar siring success on M. lewisii flowers. The observed 99.915% occurrence of parental M. lewisii and M. cardinalis in seeds collected from a sympatric population is nearly identical to that expected, based upon our field observations of pollinator behavior and our laboratory experiments of pollen competition. F1 hybrids exhibited reduced germination rates, high survivorship and reproduction, and low pollen and ovule fertility. In aggregate, the studied reproductive barriers prevent, on average, 99.87% of gene flow, with most reproductive isolation occurring prior to hybrid formation. Our results suggest that ecological factors resulting from adaptive divergence are the primary isolating barriers in this system. Additional studies of taxa at varying degrees of evolutionary divergence are needed to identify the relative importance of pre- and postzygotic isolating mechanisms in speciation.     

Sexual reproduction in the cork oak (Quercus suber L). II. Crossing intra- and interspecific barriers

Intraspecific barriers promote outcrossing while interspecific mechanisms may contribute to the isolation of species; both control the exchange of genes between plants. In this paper we establish that post-pollination mechanisms promote outcross and act at different temporal and spatial levels to control seed set and quality in Quercus species. Controlled pollinations were performed to investigate intraspecific crossing barriers in Q. suber and pollen-pistil interactions following interspecific pollinations with some simpatric Quercus species. Cytological data of intraspecific crosses in Q. suber and Q. ilex have shown different kinetics on pollen tube growth after self and outcross pollination, but pollen tubes were able to reach the base of the styles in both species and seed set was successful. Although some pre-zygotic interaction is occurring at the style, the most important interaction takes place at ovary. The cross between Q. ilex and Q. suber is possible only in one direction, revealing a case of unilateral incongruity. We show that the lack of seed set observed in the cross Q. suber×Q. ilex is due to the inability of pollen tubes to penetrate the transmitting tissue after germination. With Q. suber mainly as female parent, pollinations with other simpatric Quercus species have shown different levels of constraint on pollen tube progression at stigma and style levels. Eventual hybridisation between Quercus species will depend on the compatibility of pollen-pistil interactions, on the competitive ability of pollen genotypes, and, most important, on the overlapping of geographical, phenological and ecological factors. Differences in seed set and seed allocation was evident in all crosses, and was particularly outstanding in interspecific and in self intraspecific crosses, determining the ultimate level of seed production and quality in Quercus species. Received: 4 April 2001 / Accepted: 8 May 2001     

Reproductive ecology of a cleistogamous annual,Impatiens noli-tangere L., occurring under different environmental conditions

Demographic and reproductive schedules were compared among five populations of a cleistogamous annual,Impatients noli-tangere L., occurring in habitats with contrasting moisture and/or light environments. In all populations, flowering extended for 2–3 months during which light environment and mortality changed. Seasonal patterns of growth and mortality were significantly different among the five populations studied. The beginning and duration of flowering and the ratio of chasmogamous flowers to cleistogamous ones were also significantly different among populations. An experiment was conducted under different light conditions (open and closed) to separate the genetic and environmental components of the variation in reproductive traits observed among populations ofI. noli-tangere. Transplanted plants showed significant among-population variation in flowering time, as is observed in natural populations, suggesting genetic differentiation among populations of this species. On the other hand, the ratio of chasmogamous flowers did not differ among plants transplanted from three populations. Based on these results, the authors suggest that facultative cleistogamy is a conditional strategy under seasonally changing environments.     

INTERSPECIFIC GENE FLOW IN CUCURBITA: C. TEXANA VS. C. PEPO

The potential for interspecific genetic exchange was examined by monitoring flowering patterns, pollinator movement, and gene flow among experimental populations of the Texas gourd (Cucurbita texana) and cultivars of Cucurbita pepo. While flowering patterns and pollinator movement tended to maximize self-pollination and local gene exchange, movement of effective pollen exceeded 1,300 m. This movement, mediated by the solitary bee Xenoglossa strenua and monitored by tracking allozyme variants, produced interspecific hybrids in 5% of the progeny from experimental plants. Interspecific gene exchange occurred in either direction with either species serving as staminate or pistillate parent. No obvious constraints to gene flow among plants representing C. texana and distinctive cultivars (vars. ovifera, medullosa, melopepo) of C. pepo were detected. Genetic exchange among different species and cultivars is enhanced by the foraging behavior of Xenoglossa. Multiple visits to either staminate (pollen carryover) or pistillate (multiple pollinations) flowers often result in the deposition of mixed pollen on receptive stigmas. The wild type (C. texana) can donate and receive effective pollen when growing under both weedy and natural conditions. The observed lack of interspecific reproductive isolation supports treatment of cultivars and wild types as a single species and, in conjunction with available data concerning temporal/geographical relationships among bees, squash, gourds, and humans in eastern North America, suggests the possibility of long-term genetic interaction between wild types and domesticates.     

How chromosomal rearrangements shape adaptation and speciation: Case studies in Drosophila pseudoobscura and its sibling species Drosophila persimilis

The gene arrangements of Drosophila have played a prominent role in the history of evolutionary biology from the original quantification of genetic diversity to current studies of the mechanisms for the origin and establishment of new inversion mutations within populations and their subsequent fixation between species supporting reproductive barriers. This review examines the genetic causes and consequences of inversions as recombination suppressors and the role that recombination suppression plays in establishing inversions in populations as they are involved in adaptation within heterogeneous environments. This often results in the formation of clines of gene arrangement frequencies among populations. Recombination suppression leads to the differentiation of the gene arrangements which may accelerate the accumulation of fixed genetic differences among populations. If these fixed mutations cause incompatibilities, then inversions pose important reproductive barriers between species. This review uses the evolution of inversions in Drosophila pseudoobscura and D. persimilis as a case study for how inversions originate, establish and contribute to the evolution of reproductive isolation.     

Strength and variability of postmating reproductive isolating barriers between four European white oak species

The identification and quantification of the relative importance of reproductive isolating barriers is of fundamental importance to understand species maintenance in the face of interspecific gene flow between hybridising species. Yet, such assessments require extensive experimental fertilisations that are particularly difficult when dealing with more than two hybridising and long-generation-time species such as oaks. Here, we quantify the relative contribution of four postmating reproductive isolating barriers consisting of two prezygotic barriers (gametic incompatibility, conspecific pollen precedence) and two postzygotic barriers (germination rate, early survival) from extensively controlled pollinations between four oak species (Quercus robur, Quercus petraea, Quercus pubescens and Quercus pyrenaica) that have been shown to frequently hybridise in natural populations. We found high variation in the strength of total reproductive isolation between species, ranging from total reproductive isolation to advantage toward hybrid formation. As previously found, Q. robur pollen was unable to fertilise Q. petraea due to a strong reproductive isolating mechanism. On the contrary, Q. pubescens pollen was more efficient at fertilising Q. petraea than conspecific pollen. Overall, prezygotic barriers contribute far more than postzygotic barriers to isolate species reproductively, suggesting a role for reinforcement in the development of prezygotic barriers. Conspecific pollen precedence reduced hybrid formation when pollen competition was allowed; however, presence of conspecific pollen did not totally prevent hybridization. Our results suggest that pollen competition depends on multiple ecological and environmental parameters, including species local abundance, and that it may be of uppermost importance to understand interspecific gene flow among natural multispecies populations.     

No evidence of intrinsic reproductive isolation between two reciprocally non-monophyletic,ecologically differentiated mountain plants at an early stage of speciation

Adaptation to dissimilar habitats can trigger phenotypic and genetic differences between populations, which may, in the absence of gene flow, ultimately lead to ecological speciation. Reproductive isolation of diverging populations is a critical step at the onset of speciation. An excellent example for exploring the extent of reproductive isolation at early stages of speciation is provided by Heliosperma pusillum and H. veselskyi (Caryophyllaceae), two reciprocally non-monophyletic, ecologically differentiated species from the Alps. Interspecific gene flow—as revealed by recent genetic studies—is rare even between geographically close populations. Cross pollinations and fitness experiments revealed no evidence of intrinsic reproductive barriers, since fitness parameters measured under uniform conditions were not lower in inter- than in intraspecific crosses. Further, morphometric analyses of the offspring clearly showed that the differentiation of parental species is heritable. As parental phenotypes are likely adaptive, the intermediate morphology of hybrids may lead to reduced hybrid fitness in parental habitats. Altogether, H. pusillum and H. veselskyi provide an increasingly well characterised model system offering exciting insights into early stages of ecological speciation.     

Fertile allotetraploid from the cross betweenPhaseolus coccineus L. andPhaseolus acutifolius A. Gray

Summary As somatic hybridization and genetic transformation are not yet applicable to beans, a programme of hybridization between a male sterile line ofP. coccineus and a wild genotype ofP. acutifolius var.tenuifolius was carried out in order to introduce useful characters from the wild parent into the genome of the cultivated species. This interspecific cross is of particular interest sinceP. acutifolius is a source of resistance to many diseases, drought and high temperature. The difficulties in producing these hybrids were overcome by repeated pollinations and with the help of embryo culture. The F₁ hybrid shows a high sterility which may be explained by the poor pollen quality and the presence of a chromosomic asynapsis at meiosis. Fertile allotetraploids (Co) were successfully produced in progeny from colchicine treated cuttings of F₁ hybrids. Several (C₂) mature seeds were harvested from selfed allotetraploid plants.Abbreviations Co initial allotetraploid plants - C₁, C₂ first and second allotetraploid generation - PMC pollen mother cell     

REPRODUCTIVE ISOLATION BETWEEN SYMPATRIC RACES OF PEA APHIDS. I. GENE FLOW RESTRICTION AND HABITAT CHOICE

Determining the extent and causes of barriers to gene flow between genetically divergent populations or races of single species is an important complement to post facto analyses of the causes of reproductive isolation between recognized species. Sympatric populations of pea aphids (Acyrthosiphon pisum Harris, Homoptera: Aphididae) on alfalfa and red clover are highly genetically divergent and locally adapted. Here, hierarchical estimates of population structure based on F_st suggest that gene exchange between closely adjacent aphid populations on the two hosts is highly restricted relative to that among fields of the same host plant. Although these host-associated races are presently considered to be the same subspecies, they appear to be significantly reproductively isolated, suggesting incipient speciation. Habitat (host) choice was investigated as the first in a temporal series of factors that could reduce gene exchange between these sympatric populations. Field studies of winged colonists to newly planted fields of each host suggest pronounced habitat fidelity. This result was verified using replicated observations of the host choice behavior of different aphid genotypes for which the relative demographic performance on each host was known. These laboratory observations of behavior revealed a strong genetic correlation between habitat choice (or acceptance) and the relative performance in each habitat. Because mating occurs on the host plant, habitat choice in this system leads to assortative mating and is therefore a major cause of reproductive isolation between the sympatric pea aphid populations on alfalfa and clover. However, the extent of dispersal between hosts estimated from the field study of winged colonists (9–11%) is too great to be consistent with the genetic divergence estimated between the races. This suggests that barriers to gene flow other than host choice also exist, such as selection against migrants or hybrids in the parental environments, hybrid sterility, or hybrid breakdown.     

Genetic structure of populations and natural hybridization between <Emphasis Type="Italic">Dactylorhiza salina</Emphasis> and <Emphasis Type="Italic">D. incarnata</Emphasis> (Orchidaceae)

The results of studying the polymorphism and genetic structure of populations of D. salina and D. incarnata growing in Zabaykalsky krai and Buryatia are represented according to the data of allozyme analysis of eight genetic loci (PGI, NADHD, SKDH, GDH, PGM, DIA, ADH, and IDH). The specificity of the allelic structure of loci SKDH, PGM, and IDH is established, for which D. salina and D. incarnata reliably differ from each other. It is shown that interspecies introgressive hybrid complexes with different genetic structures were formed in Transbaikalia. Places of mass growth of D. incarnata were observed to have single plants of D. salina, the interspecies hybrids of the first and subsequent generations. Places of mass growth of D. salina were observed to contain only the hybrids that are not hybrids of the first generation. They were heterozygous not for three loci with differentiating alleles of both parents, SKDH, PGM, and IDH, but for only one of them. The degree of genetic differentiation among five populations of D. salina was on average 7.5% and that of D. incarnata was 7.1%, which in accordance with Wright’s estimation relates to mean values. The average value of FST for all studied populations of the two related species of the genus Dactylorhiza was 0.478, indicating a very high degree of genetic differentiation between D. salina and D. incarnata growing in Transbaikalia. The greatest differences between the species are for the allelic structure of loci SKDH, PGM, and IDH (FST was equal to 0.705, 0.976, and 0.762, respectively). Analysis of molecular variance (AMOVA) showed that populations of D. salina and D. incarnata in the zone where their ranges in Zabaykalsky krai and Buryatya overlap have essential differences both for the variation of alleles frequencies of eight loci (71%, d.f. = 9) and for the variability of genotypes (61%, d.f. = 9). Despite the fact that D. salina and D. incarnata explicitly share a gene flow as a result of interspecies hybridization, the genetic differentiation of populations of these related species remains at a high level.     

Population differentiation in floral and life history traits ofOxalis corniculata L. (Oxalidaceae) with style length polymorphism

A quantitative genetic analysis was conducted on the amounts and distribution of variation of 20 floral, reproductive and life history traits of a self-compatible perennial,Oxalis cornlculata L. (Oxalidaceae). This species comprises two floral morphs, homostyled and long-styled, with different breeding systems. The hierarchical design of the experiments on three homostyled and three long-styled populations allowed partitioning of variation into four levels of organization using nested ANOVAs. Seven of the 20 traits examined were differentiated between homostyled and long-styled populations. Significant genetic variance components were detected in the major traits (114 of the 120 traits) examined for six populations. Average values of variance components among families within a population across 20 traits for homostyled populations were higher than those of long-styled populations. These responses likely reflect the consequences of different levels of selfing and/or mixed mating on genetic variation in the two floral morphs ofO. corniculata. Pearson product-moment correlations for family means of seven traits selected were also calculated. Two groups of trait combinations (i.e., between floral traits, and between reproductive and life history traits) showed significant family mean correlation coefficients. The origins of these varlation patterns found in different populations ofO. corniculata are discussed in terms of the underlying selective regimes operating in each population.     

Reproductive isolation and hybridization in sympatric populations of three Dactylorhiza species (Orchidaceae) with different ploidy levels

Background and Aims

The potential for gene exchange between species with different ploidy levels has long been recognized, but only a few studies have tested this hypothesis in situ and most of them focused on not more than two co-occurring species. In this study, we examined hybridization patterns in two sites containing three species of the genus Dactylorhiza (diploid D. incarnata and D. fuchsii and their allotetraploid derivative D. praetermissa).

Methods

To compare the strength of reproductive barriers between diploid species, and between diploid and tetraploid species, crossing experiments were combined with morphometric and molecular analyses using amplified fragment length polymorphism markers, whereas flow cytometric analyses were used to verify the hybrid origin of putative hybrids.

Key Results

In both sites, extensive hybridization was observed, indicating that gene flow between species is possible within the investigated populations. Bayesian assignment analyses indicated that the majority of hybrids were F₁ hybrids, but in some cases triple hybrids (hybrids with three species as parents) were observed, suggesting secondary gene flow. Crossing experiments showed that only crosses between pure species yielded a high percentage of viable seeds. When hybrids were involved as either pollen-receptor or pollen-donor, almost no viable seeds were formed, indicating strong post-zygotic reproductive isolation and high sterility.

Conclusions

Strong post-mating reproductive barriers prevent local breakdown of species boundaries in Dactylorhiza despite frequent hybridization between parental species. However, the presence of triple hybrids indicates that in some cases hybridization may extend the F₁ generation.     

Unmasking cryptic species: morphometric and phylogenetic analyses of the Ibero‐North African Linaria incarnata complex

Linaria incarnata has been treated as a widely distributed Ibero‐North African species in the latest taxonomic reviews. Morphological and phylogenetic disparity between populations of this species has been previously reported. Here we present new morphological and phylogenetic evidence for the disintegration of L. incarnata into three distinct species: L. incarnata from the western Iberian Peninsula; L . mamorensis sp. nov. from north‐western Morocco; and L. onubensis from south‐western Spain. The relatively poor morphological differentiation between these taxa (which can be regarded as cryptic species) and their distinct phylogenetic positions indicate that characters of the L. incarnata morphotype have been acquired multiple times in the evolution of Linaria section Versicolores. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 00, 000–000.