Mechanisms Of Speciation Research Articles

Nuclear phylogeography reveals strong impacts of gene flow in big brown bats

AbstractAimUnderstanding speciation mechanisms requires disentangling processes that promote and erode population‐level divergence. Three hypotheses are raised that contemporary population structure is mainly shaped by refugial isolation, gene flow or both. Testing these hypotheses requires range‐wide phylogeography and integrative analyses across scales. Here we aimed to (1) re‐estimate the previously unresolved nuclear divergence within a widespread bat; (2) test the above three phylogeographical hypotheses and (3) inform conservation management under climatic change.LocationNorth America including the Caribbean.TaxonThe big brown bat (Eptesicus fuscus).MethodsWe collected range‐wide samples and genome‐wide markers using restriction site‐associated DNA sequencing. Population structure was analysed by clustering methods and spatial estimations. Nuclear phylogeographical divergence was estimated using tree methods (concatenation and coalescence) and network analyses (TreeMix). Phylogeographical hypotheses were tested by comparing alternative evolutionary scenarios using demographic modelling. Species distribution modelling was used to help identify Pleistocene refugia and predict future range shifts under climatic change.ResultsWe identified three populations in the Caribbean, eastern and western North America. The western population further split into three phylogeographical clades: Pacific, southwestern North America and Mexico. Discordance among mitochondrial and nuclear topologies reflected strong impacts of gene flow without sex bias. Demographic modelling supported scenarios of historical isolation followed by secondary gene flow and estimated Holocene divergence times. Species distribution was essentially continuous during glaciation with possible regional isolation, and northward range shifts were predicted under future climatic change.Main ConclusionsContemporary population divergence of big brown bats was shaped by both historical isolation and secondary gene flow, supporting the third phylogeographical hypothesis. While climatic change likely triggered initial divergence, ongoing gene flow has largely impacted the dynamic within‐species evolution and generated population divergence without speciation.

Pollinator sharing, copollination, and speciation by host shifting among six closely related dioecious fig species

The obligate pollination mutualism between figs (Ficus, Moraceae) and pollinator wasps (Agaonidae, Hymenoptera) is a classic example of cospeciation. However, examples of phylogenetic incongruencies between figs and their pollinators suggest that pollinators may speciate by host shifting. To investigate the mechanism of speciation by host shifting, we examined the phylogenetic relationships and population genetic structures of six closely related fig species and their pollinators from southern China and Taiwan-Ryukyu islands using various molecular markers. The results revealed 1) an extraordinary case of pollinator sharing, in which five distinct fig species share a single pollinator species in southern China; 2) two types of copollination, namely, sympatric copollination by pollinator duplication or pollinator migration, and allopatric copollination by host migration and new pollinator acquisition; 3) fig species from southern China have colonized Taiwan repeatedly and one of these events has been followed by host shifting, reestablishment of host specificity, and pollinator speciation, in order. Based on our results, we propose a model for pollinator speciation by host shifting in which the reestablishment of host-specificity plays a central role in the speciation process. These findings provide important insights into understanding the mechanisms underlying pollinator speciation and host specificity in obligate pollination mutualism.

Emergent Diversity and Persistent Turnover in Evolving Microbial Cross-Feeding Networks.

A distinguishing feature of many ecological networks in the microbial realm is the diversity of substrates that could potentially serve as energy sources for microbial consumers. The microorganisms are themselves the agents of compound diversification via metabolite excretion or overflow metabolism. It has been suggested that the emerging richness of different substrates is an important condition for the immense biological diversity in microbial ecosystems. In this work, we study how complex cross-feeding networks (CFN) of microbial species may develop from a simple initial community given some elemental evolutionary mechanisms of resource-dependent speciation and extinctions using a network flow model. We report results of several numerical experiments and report an in-depth analysis of the evolutionary dynamics. We find that even in stable environments, the system is subject to persisting turnover, indicating an ongoing co-evolution. Further, we compare the impact of different parameters, such as the ratio of mineralization, as well as the metabolic versatility and variability on the evolving community structure. The results imply that high microbial and molecular diversity is an emergent property of evolution in cross-feeding networks, which affects transformation and accumulation of substrates in natural systems, such as soils and oceans, with potential relevance to biotechnological applications.

Patterns of genomic divergence in sympatric and allopatric speciation of three Mihoutao (Actinidia) species.

Isolation by geographic distance is involved in the formation of potential genomic islands and the divergence of genomes, which often result in speciation. The mechanisms of sympatric and allopatric speciation associated with geographic distance remain a topic of interest to evolutionary biologists. Here, we examined genomic divergence in three Actinidia species from large-scale sympatric and allopatric regions. Genome sequence data revealed that hexaploid Actinidia deliciosa originated from Actinidia chinensis and supported the speciation-with-gene-flow model in sympatric regions. The common ancestor of Actinidia setosa and A. deliciosa migrated from the mainland to the Taiwan Island ~2.91 Mya and formed A. setosa ~0.92 Mya, and the speciation of A. setosa is consistent with the divergence-after-speciation model with selective sweeps. Geographic isolation resulted in population contraction and accelerated the process of lineage sorting and speciation due to natural selection. Genomic islands contained genes associated with organ development, local adaptation, and stress resistance, indicating selective sweeps on a specific set of traits. Our results highlight the patterns of genomic divergence in sympatric and allopatric speciation, with the mediation of geographic isolation in the formation of genomic islands during Actinidia speciation.

Intraspecific C-value variation and the outcomes in Psidium cattleyanum Sabine essential oil.

Polyploidy, a numerical alteration of the karyotype, is one of the most important mechanisms in plant speciation and diversification, but could also be detected among populations, the cytotypes. For example, Psidium cattleyanum, a polyploid complex, has chromosome numbers ranging from 2n=3x=33 to 2n=12x=132. Polyploidization causes an increase in DNA content, and both modifications may cause alteration in plant growth, physiology, and epigenetics. Based on this possibility, here we aim to verify the influence of the polyploidization on the production of P. cattleyanum essential oil chemotypes. Differences in the DNA contents, as a proxy to different ploidies, were observed and three distinct chemotypes were identified through the chromatographic profile analysis. The Psidium cattleyanum DNA content and qualitative and quantitative characteristics of the essential oils presented a positive relationship. Plants with higher DNA contents presented higher levels of oil production, which was mostly composed of hydrogenated sesquiterpenes, while plants with lower DNA contents produced lower amount of oil, which was mostly composed of hydrogenated monoterpenes. Based on the importance of essential oils, polyploid plants, which present higher DNA content, are recommended as possible matrices for the propagation of new plants with the potential to produce major compounds of agronomic and pharmacological interest.

Speciation of chromium aqua and chloro complexes in hydrochloric acid solutions at 298 K.

The distribution of metal aqua and chloro complexes is fundamental information for analysis of a chemical reaction involving these complexes. The present study investigated the speciation and structures of chromium aqua and chloro complexes using the thermodynamic model fitting analysis of UV-vis/X-ray absorption spectra. The existence of a negatively charged species was examined by adsorbability of chromium species on a strong base anion exchanger. In the results, a planar square [CrIII(H2O)4]3+, a planar square or distorted tetrahedral [CrIIICl(H2O)3]2+, a trigonal bipyramidal [CrIIICl3(H2O)2]0, and a distorted octahedral [CrIIICl4(H2O)2]- were confirmed and the thermodynamic parameters of complexation reactions were quantitatively determined. Investigation of the evolution of speciation of chromium aqua and chloro complexes in a pH 1 solution suggested the existence of [CrIIICl2(H2O) m ]+ (m = 2 or 3) during the hydration process, which diminished in the equilibrium state. The kinetic analysis deserves further investigation to understand the speciation process quantitatively.

The Neuroptera of São Tomé e Principe

Two new entomological surveys were carried out in the República Democrática de São Tomé e Principe in 2019 (earlier surveys were in 1956 and 2001). Of 16 species of Neuroptera identified, only one, a Mantispidae, appears endemic to the archipelago. Chrysopidae and Hemerobiidae account for the majority of species and individuals collected, concentrated here in plantations and anthropized environments, and known to be widely distributed in Africa and even worldwide. Unusually, the family Coniopterygidae is again absent, this time on natural terrains, confirming its earlier absence in 2001 on anthropized and plantation terrains. The findings and ecological distribution support the hypothesis that Hemerobiidae and Chrysopidae were introduced with cash crop cultivation, some of them as late as the 19th century. Their isolation in island environments is probably too recent to have allowed speciation mechanisms to generate endemic species in São Tomé.

Saccharomyces yeast hybrids on the rise.

Saccharomyces hybrid yeasts are receiving increasing attention as a powerful model system to understand adaptation to environmental stress and speciation mechanisms, using experimental evolution and omics techniques. We compiled all genomic resources available from public repositories of the eight recognized Saccharomyces species and their interspecific hybrids. We present the newest numbers on genomes sequenced, assemblies, annotations, and sequencing runs, and an updated species phylogeny using orthogroup inference. While genomic resources are highly skewed towards Saccharomyces cerevisiae, there is a noticeable movement to use wild, recently discovered yeast species in recent years. To illustrate the degree and potential causes of reproductive isolation, we reanalyzed published data on hybrid spore viabilities across the entire genus and tested for the role of genetic, geographic, and ecological divergence within and between species (28 cross types and 371 independent crosses). Hybrid viability generally decreased with parental genetic distance likely due to antirecombination and negative epistasis, but notable exceptions emphasize the importance of strain-specific structural variation and ploidy differences. Surprisingly, the viability of crosses within species varied widely, from near reproductive isolation to near-perfect viability. Geographic and ecological origins of the parents predicted cross viability to an extent, but with certain caveats. Finally, we highlight publication trends in the field and point out areas of special interest, where hybrid yeasts are particularly promising for innovation through research and development, and experimental evolution and fermentation.

The speciation view: Disentangling multiple causes of adaptive and non‐adaptive radiation in terms of speciation

AbstractBiological diversification often includes burst of lineage splitting. Such “radiation” has been known to act as evolutionary arenas with the potential to generate unique phylogenetic clusters and further novel groups. Although these radiations when accompanied by ecological diversification, so‐called “adaptive radiation” have persisted as a central premise in evolutionary biology, the ecological and genetic mechanism of such rapid diversification has remained unclear. There are several critical definitions for the pattern of adaptive radiation, and those provide delimitation of adaptive and non‐adaptive radiation. That being said, only a few studies have provided any clear demarcations in our understanding of the adaptive and non‐adaptive causes of radiation from the mechanism of speciation. Here, we review the current consensus for the causes of adaptive radiation, especially along with the recent theoretical synthesis of “ecological speciation.” Further, we suggest the signature of adaptive and non‐adaptive radiation in the earliest stages of diversification from the viewpoint of speciation. These criteria from the speciation view are useful to find the cases with the signatures of adaptive/non‐adaptive radiation.

植物交配系统与分子进化理论及其应用研究

<p indent="0mm">With the transition of plant mating system from outcrossing to selfing, both the flower traits and genomic structure develop into selfing syndromes. Here we systematically discussed the theoretical basis and the molecular mechanisms that form these characteristics, and analyzed the basic theory of the interactions between mating system and different evolutionary processes. Selfing results in decrease in the effective population size, mutation rate, recombination rate and selection efficacy, but increase in genetic drift effects and linkage disequilibrium. These theories are effective in explaining the patterns of the relative rate of nonsynonymous to synonymous mutations, the change of the number of transposable elements, the genetic correlation between multigene family members and the pattern of genomic structure. Although molecular markers are often employed to score mating system, studies of mating system and molecular evolution have been mainly in different research fields. Some questions of biological significance could arise from an integration of these two research fields, such as the mode of driving or reinforcing molecular mechanism of speciation that the mating system operates, the different models of molecular evolution between populations of selfing versus outcrossing system, and the distinct genomic structure between populations of the same species with different mating systems. Clarification of these questions aids in our understanding of the role that mating system plays in affecting molecular evolution within and between species.

Ecological Divergence and the History of Gene Flow in the Nearctic Milksnakes (Lampropeltis triangulum Complex).

Many phylogeographic studies on species with large ranges have found genetic-geographic structure associated with changes in habitat and physical barriers preventing or reducing gene flow. These interactions with geographic space, contemporary and historical climate, and biogeographic barriers have complex effects on contemporary population genetic structure and processes of speciation. While allopatric speciation at biogeographic barriers is considered the primary mechanism for generating species, more recently it has been shown that parapatric modes of divergence may be equally or even more common. With genomic data and better modeling capabilities, we can more clearly define causes of speciation in relation to biogeography and migration between lineages, the location of hybrid zones with respect to the ecology of parental lineages, and differential introgression of genes between taxa. Here, we examine the origins of three Nearctic milksnakes (Lampropeltis elapsoides, Lampropeltis triangulum and Lampropeltis gentilis) using genome-scale data to better understand species diversification. Results from artificial neural networks show that a mix of a strong biogeographic barrier, environmental changes, and physical space has affected genetic structure in these taxa. These results underscore conspicuous environmental changes that occur as the sister taxa L. triangulum and L. gentilis diverged near the Great Plains into the forested regions of the Eastern Nearctic. This area has been recognized as a region for turnover for many vertebrate species, but as we show here the contemporary boundary does not isolate these sister species. These two species likely formed in the mid-Pleistocene and have remained partially reproductively isolated over much of this time, showing differential introgression of loci. We also demonstrate that when L. triangulum and L. gentilis are each in contact with the much older L. elapsoides, some limited gene flow has occurred. Given the strong agreement between nuclear and mtDNA genomes, along with estimates of ecological niche, we suggest that all three lineages should continue to be recognized as unique species. Furthermore, this work emphasizes the importance of considering complex modes of divergence and differential allelic introgression over a complex landscape when testing mechanisms of speciation. [Cline; delimitation; Eastern Nearctic; Great Plains; hybrids; introgression; speciation.].

Phylogenomic analyses of the East Asian endemic Abelia (Caprifoliaceae) shed insights into the temporal and spatial diversification history with widespread hybridization.

Abelia (Caprifoliaceae) is a small genus with five species, including one artificial hybrid and several natural hybrids. The genus has a discontinuous distribution in Mainland China, Taiwan Island and the Ryukyu Islands, providing a model system to explore the mechanisms of species dispersal in the East Asian flora. However, the current phylogenetic relationships within Abelia remain uncertain. We reconstructed the phylogenetic relationships within Abelia using nuclear loci generated by target enrichment and plastomes from genome skimming. Divergence time estimation, ancestral area reconstruction and ecological niche modelling (ENM) were used to examine the diversification history of Abelia. We found extensive cytonuclear discordance across the genus. By integrating lines of evidence from molecular phylogenies, divergence times and morphology, we propose to merge Abelia macrotera var. zabelioides into A. uniflora. Network analyses suggested that there have been multiple widespread hybridization events among Abelia species. These hybridization events may have contributed to the speciation mechanism and resulted in the high observed morphological diversity. The diversification of Abelia began in the early Eocene, followed by A. chinensis var. ionandra colonizing Taiwan Island during the Middle Miocene. The ENM results suggested an expansion of climatically suitable areas during the Last Glacial Maximum and range contraction during the Last Interglacial. Disjunction between the Himalayan-Hengduan Mountain region and Taiwan Island is probably the consequence of topographical isolation and postglacial contraction. We used genomic data to reconstruct the phylogeny of Abelia and found a clear pattern of reticulate evolution in the group. In addition, our results suggest that shrinkage of postglacial range and the heterogeneity of the terrain have led to the disjunction between Mainland China and Taiwan Island. This study provides important new insights into the speciation process and taxonomy of Abelia.

Chromosome-scale genome assembly of Castanopsis tibetana provides a powerful comparative framework to study the evolution and adaptation of Fagaceae trees.

Fagaceae species are increasingly used as models to elucidate the process and mechanism of adaptation and speciation by integrating ecology, evolution and genomics. The genus Castanopsis belongs to the family Fagaceae and is mainly distributed across subtropical and tropical Asia. In the present study, we reported the first chromosome-scale genome assembly of Castanopsis tibetana, a common species of evergreen broadleaved forests in subtropical China. The combination of Nanopore sequencing and Hi-C technologies enabled a high-quality genome assembly. The final assembled genome size of C.tibetana was 878.6Mb (97.6% of the estimated genome size), consisting of 477 contigs with an N50length of 3.3Mb. The benchmarking universal single-copy orthologue (BUSCO) assessment indicated a completeness of 93.0%. Hi-C scaffolding generated 12pseudochromosomes, representing 98.7% of the assembled genome. Subsequently, 40,937 protein-coding genes were predicted and 90.04% of them were functionally annotated. More than 476.9Mb of repetitive sequences (54.3% of the genome) were identified, and the percentage of the genome covered by TE elements was 39.98%. Comparative genomics analysis revealed that C.tibetana was most closely related to Castanea mollissima and diverged at 18.48Ma, and that C.tibetana has undergone considerable gene family expansion and contraction. Evidence of positive selection was detected in 53genes, which showed different arrangement pattern compared to Quercus robur. The chromosome-scale genome assembly of C.tibetana will expand Fagaceae genome resources across the family and provide a powerful comparative framework to study the adaptation and evolution of Fagaceae trees.

The mode of speciation during a recent radiation in open-ocean phytoplankton

Despite the enormous ecological importance of marine phytoplankton, surprisingly little is known about how new phytoplankton species originate and evolve in the open ocean, in the absence of apparent geographic barriers that typically act as isolation mechanisms in speciation. To investigate the mechanism of open-ocean speciation, we combined fossil and climatic records from the late Quaternary with genome-wide evolutionary genetic analyses of speciation in the ubiquitous and abundant pelagic coccolithophore genus Gephyrocapsa (including G.huxleyi, formerly known as Emiliania huxleyi). Based on the analysis of 43 sequenced genomes, we report that the best-fitting scenario for all speciation events analyzed included an extended period of complete isolation followed by recent (Holocene) secondary contact, supporting the role of geographic or oceanographic barriers in population divergence and speciation. Consistent with this, fossil data reveal considerable diachroneity of species first occurrence. The timing of all speciation events coincided with glacial phases of glacial-interglacial cycles, suggesting that stronger isolation between the ocean basins and increased segregation of ecological niches during glaciations are important drivers of speciation in marine phytoplankton. The similarity across multiple speciation events implies the generality of this inferred speciation scenario for marine phytoplankton.

Novel wing display and divergent agonistic behaviors of two incipient Psolodesmus damselflies.

Sexual selection via male competition is a strong evolutionary force that can drive rapid changes in competitive traits and subsequently lead to population divergence and speciation. Territorial males of many odonates are known to use their colorful wings as visual signals and to perform agonistic displays toward intruders. Psolodesmus mandarinus dorothea and Psolodesmus mandarinus mandarinus are two parapatrically distributed sister damselflies that share similar ecological characteristics but differ markedly in wing coloration. The wings of P. m. dorothea are mostly clear, whereas those of P. m. mandarinus have a large area of black pigmentation and a central white patch. We investigated whether territorial males of the two damselflies at breeding sites display distinct agonistic behaviors associated with their respective wing colors. Behavioral interactions between territorial and intruder males and their wing kinematics were filmed and analyzed for P. m. dorothea in Lienhuachih of central Taiwan, and P. m. mandarinus in Tianxiyuan andFusan of northern Taiwan. We observed that the P. m. mandarinus males exhibited a novel set of perched wing displays, which was not only absent in its sister P. m. dorothea but also previously unknown in Odonata. At breeding sites, perched rival males of P. m. mandarinus with pigmented wings exhibited escalating agonistic wing-flapping and wing-hitting displays toward each other. In contrast, territorial males of P. m. dorothea with clear wings engaged only in aerial chase or face-to-face hovering when intruder males approached from the air. These results indicate that the two sister P. mandarinus damselflies diverged behaviorally in territorial contests and support the hypothesis of coadaptation on the basis of wing colors and types of wing movement in Odonata. Our findings further suggest that divergent agonistic wing displays may play a pivotal role in the speciation mechanism of P. mandarinus damselflies. The sequential analyses of behavioral characteristics and progression suggest that P. m. mandarinus damselflies likely use mutual assessment of rivals in territorial contests.

Phylogeographic reconstruction of the marbled crayfish origin

The marbled crayfish (Procambarus virginalis) is a triploid and parthenogenetic freshwater crayfish species that has colonized diverse habitats around the world. Previous studies suggested that the clonal marbled crayfish population descended as recently as 25 years ago from a single specimen of P. fallax, the sexually reproducing parent species. However, the genetic, phylogeographic, and mechanistic origins of the species have remained enigmatic. We have now constructed a new genome assembly for P. virginalis to support a detailed phylogeographic analysis of the diploid parent species, Procambarus fallax. Our results strongly suggest that both parental haplotypes of P. virginalis were inherited from the Everglades subpopulation of P. fallax. Comprehensive whole-genome sequencing also detected triploid specimens in the same subpopulation, which either represent evolutionarily important intermediate genotypes or independent parthenogenetic lineages arising among the sexual parent population. Our findings thus clarify the geographic origin of the marbled crayfish and identify potential mechanisms of parthenogenetic speciation.

Stable allopolyploidy in a hybrid zone within the Matsukawaura Lagoon (Japan) between Neopyropia yezoensis and Neopyropia tenera (Bangiales, Rhodophyta) and its relevance to speciation mechanisms and crop improvement

AbstractTo confirm the distribution of the allopolyploid and ancestral species in the Matsukawaura Lagoon in a hybrid zone between Neopyropia yezoensis and Neopyropia tenera, molecular marker analyses and culture experiments were performed using collected blades. All the blades examined, except for one blade identified as N. tenera, exhibited nuclear genotypes of both species, but only the chloroplast haplotype of N. yezoensis. In addition, four conchocelis strains were isolated from each of the blades, and they contained genotypes of both N. yezoensis and N. tenera; a conchocelis strain from the sole N. tenera blade was also isolated. Using the conchocelis strains, F1 blades that were produced from archeospores were obtained under laboratory culture and examined by nuclear DNA markers. These results indicated that the collected blades and the examined F1 blades, except for the single N. tenera blade, were allodiploids and that the four conchocelis strains could be allotetraploids derived from interspecific hybridization between N. yezoensis as the maternal strain and N. tenera as the paternal strain. Therefore, most of the Neopyropia blades growing in the Matsukawaura Lagoon are presumed to be from allopolyploid lineages derived from interspecific hybridization. Furthermore, a growth comparison between the allopolyploid strain and the N. tenera strain was carried out under identical culture conditions. The rate of increase in blade length was significantly higher in the allopolyploid hybrid than in the pure N. tenera. The results suggest that further exploration of allopolyploidy in the potential hybrid zones will be needed not only for the investigation of speciation mechanisms in the bladed Bangiales but also for the genetic improvement of the marine crop.

Divergence and hybridization in sea turtles: Inferences from genome data show evidence of ancient gene flow between species.

Reconstructing past events of hybridization and population size changes are required to understand speciation mechanisms and current patterns of genetic diversity, and ultimately contribute to species' conservation. Sea turtles are ancient species currently facing anthropogenic threats including climate change, fisheries, and illegal hunting. Five of the seven extant sea turtle species are known to currently hybridize, especially along the Brazilian coast where some populations can have ~32%–42% of hybrids. Although frequently observed today, it is not clear what role hybridization plays in the evolutionary diversification of this group of reptiles. In this study, we generated whole genome resequencing data of the five globally distributed sea turtle species to estimate a calibrated phylogeny and the population size dynamics, and to understand the role of hybridization in shaping the genomes of these ancient species. Our results reveal discordant species divergence dates between mitochondrial and nuclear genomes, with a high frequency of conflicting trees throughout the nuclear genome suggesting that some sea turtle species frequently hybridized in the past. The reconstruction of the species' demography showed a general decline in effective population sizes with no signs of recovery, except for the leatherback sea turtle. Furthermore, we discuss the influence of reference bias in our estimates. We show long‐lasting ancestral gene flow events within Chelonioidea that continued for millions of years after initial divergence. Speciation with gene flow is a common pattern in marine species, and it raises questions whether current hybridization events should be considered as a part of these species' evolutionary history or a conservation issue.

Female mate preferences do not predict male sexual signals across populations

Abstract New species can arise when female preferences and male sexual signals diverge across populations and thereby reduce mating between populations. Under this hypothesized mechanism for speciation, mate preferences and sexual signals should be correlated, but divergent, across populations. We evaluated this prediction using spadefoot toads (Spea multiplicata). We measured a sexually selected male signal (call rate) for which female preferences are known to vary across populations in response to the risk of hybridizing with another species. Contrary to expectation, we found no correlation between male call rate and female preferences across populations. We discuss possible mechanisms of this pattern, including the effect of gene flow from heterospecifics on male call rate. Our results suggest that, even when populations vary in mating traits, the independent evolution of female preferences and male sexual signals might impede reproductive isolation between populations.

Variation in heteroploid reproduction and gene flow across a polyploid complex: One size does not fit all.

Whole‐genome duplication is considered an important speciation mechanism in plants. However, its effect on reproductive isolation between higher cytotypes is not well understood. We used backcrosses between different ploidy levels and surveys of mixed‐ploidy contact zones to determine how reproductive barriers differed with cytotype across a polyploid complex. We backcrossed F1 hybrids derived from 2X‐4X and 4X‐6X crosses in the Campanula rotundifolia autopolyploid complex, measured backcross fitness, and estimated backcross DNA cytotype. We then sampled four natural mixed‐ploidy contact zones (two 2X‐4X and two 4X‐6X), estimated ploidy, and genotyped individuals across each contact zone. Reproductive success and capacity for gene flow was markedly lower for 2X‐4X than 4X‐6X hybrids. In fact, 3X hybrids could not backcross; all 2X‐4X backcross progeny resulted from neotetraploid F1 hybrids. Further, no 3X individuals were found in 2X‐4X contact zones, and 2X and 4X individuals were genetically distinct. By contrast, backcrosses of 5X hybrids were relatively successful, particularly when crossed to 6X individuals. In 4X‐6X contact zones, 5X individuals and aneuploids were common and all cytotypes were largely genetically similar and spatially intermixed. Taken together, these results provide strong evidence that reproduction is low between 2X and 4X cytotypes, primarily occurring via unreduced gamete production, but that reproduction and gene flow are ongoing between 4X and 6X cytotypes. Further, it suggests whole‐genome duplication can result in speciation between diploids and polyploids, but is less likely to create reproductive barriers between different polyploid cytotypes, resulting in two fundamentally different potentials for speciation across polyploid complexes.