Patterns of cytotype variation of <Emphasis Type="Italic">Turnera sidoides</Emphasis> subsp. <Emphasis Type="Italic">pinnatifida</Emphasis> (Turneraceae) in mountain ranges of central Argentina

Cytogeographical variability among 564 plants from 26 populations of Turnera sidoides subsp. pinnatifida in mountain ranges of central Argentina was analysed with meiotic chromosome counts and flow cytometry and is described at regional and local scales. Populations were primarily tetraploids (2n = 4x = 28), although diploid (2n = 2x = 14), hexaploid (2n = 2x = 42), and mixed populations of diploids and triploids (2n = 3x = 21) were also found. Diploids, triploids, and hexaploids were fewer in number and restricted to narrow areas, while tetraploids were the most common and geographically widespread cytotype. Diploids grew at higher altitudes and in colder and wet locations; tetraploids had the broadest ecological spectrum, while hexaploids occurred at the lowest altitudes and in drier conditions. The cytotypes were also spatially segregated at a microgeographical scale. Diploids grew in the piedmont, tetraploids were in the adjacent valley, and in the contact zone of both cytotypes, patches of diploids and triploids were found. At a regional scale, the distribution of the cytotypes may be governed by a combination of ecological and historical variables, while segregation in the contact zone may be independent of the selective environment because the cytotypes are unable to coexist as a result of reproductive exclusion. The role of triploids is also discussed.     

Photosynthetic characteristics of three ploidy levels of Allium oleraceum L. (Amaryllidaceae) differing in ecological amplitude

To elucidate the mechanisms of the adaptive advantages of polyploidy, there is a need to identify physiological traits that participate in the success of polyploids. We studied selected photosynthetic characteristics, stomatal density, and specific leaf area of three ploidy levels (2n = 4x, 5x, 6x) of the geophyte Allium oleraceum that partially differ in their ecological niches. Although the cytotypes were on average similar with regard to most of the measured photosynthetic traits, the hexaploids showed more rapid initial photosynthetic induction and a tendency for a higher maximum photosynthetic rate per unit area. The stomatal density was not affected by ploidy, though the specific leaf area was reduced for the hexaploids compared to the other cytotypes. A lower intracytotype variation was found for most of the studied photosynthetic and anatomical traits for the hexaploids compared to the large variation found within other cytotypes. A comparison of the photosynthetic traits between the cytotypes showed that the ecological differentiation between cytotypes is only weakly related to the characteristics of their photosynthetic apparatus. However, contrasting ranges of variability in the measured traits between the cytotypes can be related to previously observed differences between cytotypes with regard to the ranges of intracytotype genetic variation, genome size variation, and niche breadth. A higher variability of photosynthetic traits in tetraploids and pentaploids may be related to the existence of a spectrum of types adapted to different environmental conditions. Hexaploids may represent a recently formed cytotype adapted to open environmental conditions.     

Neopolyploidy in Spartina pectinata Link: 1. Morphological analysis of tetraploid and hexaploid plants in a mixed natural population

Prairie cordgrass has been reported as a multi-polyploidy species having three cytotypes: tetra- (2n?=?4x?=?40), hexa- (2n?=?6x?=?60), and octoploid (2n?=?8x?=?80). A mixed-ploidy population comprising tetraploids and hexaploids was recently found at a single location in Illinois. However, adaptation and morphological differences between tetra- and hexaploids occurring in natural conditions as well as the contact zones of these cytotypes have yet to be determined. In this study, the cytotypes of 147 individuals of prairie cordgrass collected across the contact zone (4x?+?6x) were determined by flow cytometry using somatic G1 nuclei, and the results were confirmed by chromosome counts. Nineteen morphological characteristics were compared between the cytotypes. Tetra- and hexaploid plants have 2C genome sizes of 1.57 and 2.36?pg with chromosome counts of 40 and 60, respectively. This increase in polyploidy resulted in a greater variability of morphological expression in Illinois prairie cordgrass. Substantial differences in the flowering time, stomatal size, and plant morphological characteristics were observed between tetra- and hexaploids. The results indicate that the increasing of ploidy level in prairie cordgrass resulted in increased plant size in ploidy mixtures. The recent event of ploidy mixtures in prairie cordgrass natural populations offers unique opportunities for studying the formation and establishment of neopolyploidy.     

A cytogenetic study of Eriophyllum confertiflorum (Compositae,Helenieae)

Meiotic analyses of 176 plants from 130 populations of Eriophyllum confertiflorum var. confertiflorum showed that diploids (n = 8) and tetraploids were about equally frequent and that hexaploids and octoploids were rare. Supernumerary chromosomes were found in 14% of the individuals; other meiotic aberrations were uncommon. Eriophyllum confertiflorum var. tanacetiflorum is octoploid. Migration from southwest North America, tolerance of aridity, and spontaneous origin of polyploids may explain the nonrandom distribution of cytotypes in E. confertiflorum var. confertiflorum. Artificial intravarietal crosses of E. confertiflorum var. confertiflorum yielded vigorous, bivalent-forming progeny with mean pollen stainabilities of 41%-96% and 88%-90% at the diploid and tetraploid levels, respectively. Tetraploid x diploid crosses gave triploids in one case and tetraploids in another. The progenies of some artificial crosses between distant populations of E. confertiflorum var. confertiflorum had significantly lower rates of germination and pollen viability.     

Sympatric diploid and hexaploid cytotypes of Senecio carniolicus (Asteraceae) in the Eastern Alps are separated along an altitudinal gradient

We explored the fine-scale distribution of cytotypes of the mountain plant Senecio carniolicus along an altitudinal transect in the Eastern Alps. Cytotypes showed a statistically significant altitudinal segregation with diploids exclusively found in the upper part of the transect, whereas diploids and hexaploids co-occurred in the lower range. Analysis of accompanying plant assemblages revealed significant differences between cytotypes along the entire transect but not within the lower part only, where both cytotypes co-occur. This suggests the presence of ecological differentiation between cytotypes with the diploid possessing the broader ecological niche. No tetraploids were detected, indicating the presence of strong crossing barriers. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The Microgeographical Patterns of Morphological and Molecular Variation of a Mixed Ploidy Population in the Species Complex Actinidia chinensis

Polyploidy and hybridization are thought to have significant impacts on both the evolution and diversification of the genus Actinidia, but the structure and patterns of morphology and molecular diversity relating to ploidy variation of wild Actinidia plants remain much less understood. Here, we examine the distribution of morphological variation and ploidy levels along geographic and environmental variables of a large mixed-ploidy population of the A. chinensis species complex. We then characterize the extent of both genetic and epigenetic diversity and differentiation exhibited between individuals of different ploidy levels. Our results showed that while there are three ploidy levels in this population, hexaploids were constituted the majority (70.3%). Individuals with different ploidy levels were microgeographically structured in relation to elevation and extent of niche disturbance. The morphological characters examined revealed clear difference between diploids and hexaploids, however tetraploids exhibited intermediate forms. Both genetic and epigenetic diversity were high but the differentiation among cytotypes was weak, suggesting extensive gene flow and/or shared ancestral variation occurred in this population even across ploidy levels. Epigenetic variation was clearly correlated with changes in altitudes, a trend of continuous genetic variation and gradual increase of epigenomic heterogeneities of individuals was also observed. Our results show that complex interactions between the locally microgeographical environment, ploidy and gene flow impact A. chinensis genetic and epigenetic variation. We posit that an increase in ploidy does not broaden the species habitat range, but rather permits A. chinensis adaptation to specific niches.     

Multiple autopolyploidizations and range expansion of Allium przewalskianum Regel. (Alliaceae) in the Qinghai‐Tibetan Plateau

We used the Allium przewalskianum diploid–tetraploid complex on the Qinghai‐Tibetan Plateau (QTP) as a model to examine how this complex responded to the Quaternary climatic oscillations, and whether multiple autopolyploidizations have occurred. We sequenced five chloroplast DNA (cpDNA) fragments (accD‐psaI, trnH‐psbA, trnL‐trnF, trnS‐trnG and rpl16‐intron) in 306 individuals (all of known ploidy level) from 48 populations across the distribution of this species complex. We identified a total of 32 haplotypes—11 in diploids only, 13 in tetraploids only, and 8 found in both cytotypes. This, plus network analyses, indicated that tetraploids have arisen independently from diploids at least eight times. Most populations in the eastern QTP contained multiple haplotypes, but only a single haplotype was found for 17 tetraploid populations on the western QTP, suggesting a recent colonization of the western QTP. We further found that this species complex underwent an earlier range expansion around 5–150 thousand years ago (kya), after the largest glacial period (800–170 kya) in the QTP. In addition, the high frequencies of tetraploids in the QTP suggested that the tetraploid A. przewalskianum cytotype has evolutionary advantages over diploids in colonizing and/or surviving the arid habitats of the QTP.     

Hybridization between diploid Centaurea pseudophrygia and tetraploid C. jacea (Asteraceae): the role of mixed pollination,unreduced gametes,and mentor effects

We studied hybridization between the diploid Centaurea pseudophrygia and the tetraploid C. jacea by performing crossing experiments and screening natural populations using flow cytometry. The experiments confirm that the studied species exhibit strong reproductive isolation. Interspecific hybrids were formed at a low frequency, including triploids (originating from reduced gametes) and tetraploids (involving unreduced gametes of the diploids). In contrast, hybrids were almost absent among seeds and adult plants of natural mixed populations and among the offspring from experimental pollinations with a mixture of pollen of both ploidy levels. We found that mixed pollination is an important mechanism for preventing hybridization between plants of different ploidy levels and sustaining the reproduction of the tetraploids. A mentor effect (induced selfing in the presence of pollen of different ploidy levels) was observed in both diploids and tetraploids, reinforcing the reproductive isolation between cytotypes. Higher ploidy levels (pentaploid, hexaploid) involving unreduced gametes of the tetraploid species were identified. Notably, pentaploids were discovered for the first time in Centaurea sect. Jacea. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 93–106.     

Agrostis mertensii (Poaceae) in hokkaido

Populations of theAgrostis flaccida-mertensii complex in Hokkaido were examined as part of the taxonomic study of this complex, which has a wide range and comprises a polyploid series of 2x–8x (x=7). The principal aim was to clarify the taxonomic status of hexaploids growing in Hokkaido. Field explorations were made in the mountains and on the coasts in Hokkaido, and the materials were gathered from 19 localities. The occurrence of four cytotypes (2x, 4x, 6x, 8x), as well as some triploids and septaploids, of this complex in Hokkaido was confirmed, and the ranges of respective cytotypes were delineated. Morphological examinations showed that diploids, triploids and tetraploids are referable toAgrostis flaccida and octoploids toA. mertensii. The hexaploids of Hokkaido were morphologically different fromA. flaccida and also from the related hexaploid species,A. tateyamensis, distributed in Honshu. The hexaploids were quite similar to octoploidA. mertensii in all the morphological features examined and also in habitat preference. Septaploids were sterile hybrids between hexaploids and octoploids. A conclusion was drawn that the hexaploid plant of Hokkaido should be grouped with the octoploid in one and the same species,Agrostis mertensii Trin.Agrostis mertensii in this amended sense is discussed from a cytogeographical viewpoint.     

Polyploidy and habitat differentiation in Dactylis glomerata L. from Galicia (Spain)

Summary The microdistribution of diploid and tetraploid plants of Dactylis glomerata L. was examined and related to their immediate environment in several sites in central Galicia, where morphologically indistinguishable individuals of both ploidies grow in sympatry. The two related cytotypes differed in habitat preference. Diploids were mainly confined to the low-density forest-floor habitat in woodlands of mostly ancient origin, whereas tetraploids were widespread in varied habitats but clearly predominant in open areas, particularly in disturbed anthropic sites. The in situ comparison of plant performance showed that where plants of each ploidy were more common they produced more tillers, panicles and seeds. This habitat preference closely reflected differences in life-history characteristics. The tetraploids had an early and short flowering time almost always completed before the aestival drought, whereas the diploids began to flower several weeks later and flowered throughout the drought. Comparisons along artificial gradients of soil water availability and light transmittance indicated that the cytotypes had distinct physiological requirements which probably originated in metabolic and more general genetic differentiation and could be directly attributable to ploidy. Habitat differentiation increases the species' colonizing ability. It also amplifies divergence in reproductive strategy between diploids and tetraploids, which reduces ineffective crossing between cytotypes and thereby permits them to coexist in sympatry. The effect of hybridization at the polyploid level on the differentiation between cytotypes was assessed from the recent introduction of a foreign tetraploid entity into the study area. Hybridization between the two distinct tetraploids was found to increase habitat differentiation between the diploids and the tetraploids, but the major part of this differentiation is probably attributable to ploidy itself.     

Polyploidy associated with altered and broader ecological niches in the Claytonia perfoliata (Portulacaceae) species complex

? Premise of the study: Polyploids are often hypothesized to have distinct and broader niches than their diploid progenitors. Differences in geographic distributions between diploid and polyploids are frequently used to infer niche differentiation and increased breadth, but they are seldom used to test these hypotheses explicitly. ? Methods: Niche overlap and breadth were compared for diploids, tetraploids, and hexaploids of three taxa in the Claytonia perfoliata complex (C. parviflora, C. perfoliata, and C. rubra) with the use of species distribution models. Resampling and randomization approaches were used to test hypotheses of niche differentiation, breadth, and conservatism. ? Key results: Niche differentiation was detected between polyploid and diploid cytotypes assigned to the same taxon (e.g., C. parviflora 2× vs. 4×) but not between hexaploids and tetraploids within a taxon (e.g., C. parviflora 4× vs. 6×). Individual hexaploid cytotypes had broader ecological niches than individual diploid cytotypes. However, as a group the three hexaploid taxa did not exceed the combined niche breadth of the three diploids, suggesting that polyploidy does not result in transgressive niche breadth for this group. Niche overlap was lowest among diploids and was highest among the three hexaploid cytotypes, consistent with introgression associated with polyploidy resulting in greater ecological similarity. Although cytotypes possessed nonidentical niches, after accounting for environmental differences among ranges, cytotypes were more similar than expected, suggesting niche conservatism and similar responses to environmental characteristics. ? Conclusions: These results suggest that polyploids occupy distinct and broader niches relative to diploids but that cytotypes also share fundamentally similar responses to environmental variation across ploidy levels.     

A CYTOGEOGRAPHIC STUDY OF CHAENACTIS DOUGLASII (COMPOSITAE,HELENIEAE)

Analysis of 512 plants derived from 200 populations shows that the widely distributed western North American Chaenactis douglasii species-complex consists of diploids (n = 6), triploids, tetraploids, and hexaploids. Microsporocytes were the source of most of the chromosome counts. About 9% of all plants examined had one or more full-sized extra chromosomes. Multivalents, usually a ring or chain of four chromosomes, were almost entirely restricted to polyploids, where one or more were identified in 38% of the tetraploids and 33% of the hexaploids. With two exceptions, diploids and polyploids were not found in the same population. Frequencies of diploid, triploid, tetraploid, and hexaploid populations were, respectively, 34, 1.5, 55 and 9.5%. With significant exceptions, diploid populations predominate in the Pacific and Rocky Mountain Systems, whereas polyploid ones are most frequent in the intervening plateaus. Ploidy level is correlated with age of substrate, rather than with climate, elevation, vegetation, or soil type. Range, morphology, ploidy level, and meiotic behavior suggest that var. achilleifolia tetraploids and hexaploids are descendents of hybrids between other variants of the complex. The diploid-tetraploid-hexaploid geographic distribution and the age of the substrates where each tends to occur suggest that the complex evolved in late Cenozoic time in response to major climatic and geologic changes that induced migration and hybridization. The hybrid derivatives, stabilized by polyploidy and tolerant of increasing aridity, came to occupy newly available habitats in areas disturbed by volcanic activity and glacial or glacial-related processes.     

DNA ploidy-level variation in native and invasive populations of Lythrum salicaria at a large geographical scale

Aim  This study aimed to document precisely the patterns of DNA ploidy variation in the native and secondary ranges of Lythrum salicaria distribution. The hypothesis that species invasiveness had been induced by a switch in ploidy level was addressed.
Location  Europe, Middle East, North America.
Methods  DNA ploidy levels of 1884 progenies of 578+ plants collected at 124 localities were determined by DAPI flow cytometry.
Results  Large cytotype variation (2 x , 3 x , 4 x and 6 x ) was found across the native area of distribution (64 populations covering 12 European and two Middle Eastern countries). DNA hexaploids were detected for the first time, and rare DNA triploids were reliably confirmed. DNA tetraploids largely prevailed across the native range studied, while DNA diploids and DNA hexaploids were recorded only in Israel and Turkey, respectively. DNA triploid progenies occurred in one population from Hungary (together with DNA tetraploids). Sympatric growth of DNA tetraploids and DNA hexaploids was repeatedly encountered in Turkey. In contrast, cytotype uniformity was a typical feature of the invasive North American plants. Sixty populations, covering 13 states of the USA and provinces of Canada, were characterized by the presence of only DNA tetraploids.
Main conclusions  Several L. salicaria cytotypes (2 x , 3 x , 4 x , 6 x ) occur in the native range of distribution, with much variation concentrated in the Middle Eastern countries, whereas only DNA tetraploids appeared to occur in North America. Our data show that the invasive spread of North American populations was not triggered by differences in ploidy level. Alternative explanations should be sought.     

Ecological differentiation,lack of hybrids involving diploids,and asymmetric gene flow between polyploids in narrow contact zones of Senecio carniolicus (syn. Jacobaea carniolica,Asteraceae)

Areas of immediate contact of different cytotypes offer a unique opportunity to study evolutionary dynamics within heteroploid species and to assess isolation mechanisms governing coexistence of cytotypes of different ploidy. The degree of reproductive isolation of cytotypes, that is, the frequency of heteroploid crosses and subsequent formation of viable and (partly) fertile hybrids, plays a crucial role for the long‐term integrity of lineages in contact zones. Here, we assessed fine‐scale distribution, spatial clustering, and ecological niches as well as patterns of gene flow in parental and hybrid cytotypes in zones of immediate contact of di‐, tetra‐, and hexaploid Senecio carniolicus (Asteraceae) in the Eastern Alps. Cytotypes were spatially separated also at the investigated microscale; the strongest spatial separation was observed for the fully interfertile tetra‐ and hexaploids. The three main cytotypes showed highly significant niche differences, which were, however, weaker than across their entire distribution ranges in the Eastern Alps. Individuals with intermediate ploidy levels were found neither in the diploid/tetraploid nor in the diploid/hexaploid contact zones indicating strong reproductive barriers. In contrast, pentaploid individuals were frequent in the tetraploid/hexaploid contact zone, albeit limited to a narrow strip in the immediate contact zone of their parental cytotypes. AFLP fingerprinting data revealed introgressive gene flow mediated by pentaploid hybrids from tetra‐ to hexaploid individuals, but not vice versa. The ecological niche of pentaploids differed significantly from that of tetraploids but not from hexaploids.     

Detection of individual ploidy levels with genotyping‐by‐sequencing (GBS) analysis

Ploidy levels sometimes vary among individuals or populations, particularly in plants. When such variation exists, accurate determination of cytotype can inform studies of ecology or trait variation and is required for population genetic analyses. Here, we propose and evaluate a statistical approach for distinguishing low‐level ploidy variants (e.g. diploids, triploids and tetraploids) based on genotyping‐by‐sequencing (GBS) data. The method infers cytotypes based on observed heterozygosity and the ratio of DNA sequences containing different alleles at thousands of heterozygous SNPs (i.e. allelic ratios). Whereas the method does not require prior information on ploidy, a reference set of samples with known ploidy can be included in the analysis if it is available. We explore the power and limitations of this method using simulated data sets and GBS data from natural populations of aspen (Populus tremuloides) known to include both diploid and triploid individuals. The proposed method was able to reliably discriminate among diploids, triploids and tetraploids in simulated data sets, and this was true for different levels of genetic diversity, inbreeding and population structure. Power and accuracy were minimally affected by low coverage (i.e. 2×), but did sometimes suffer when simulated mixtures of diploids, autotetraploids and allotetraploids were analysed. Cytotype assignments based on the proposed method closely matched those from previous microsatellite and flow cytometry data when applied to GBS data from aspen. An R package (gbs2ploidy) implementing the proposed method is available from CRAN.     

THE HORDEUM VIOLACEUM COMPLEX OF IRAN

Fifty-seven Iranian collections of Hordeum violaceum Boiss. & Huet, a perennial forage grass, contained diploid (2n = 14), tetraploid (2n = 28), and hexaploid (2n = 42) chromosome races. All collections came from moderate to high elevations in the Alborz and Zagros mountains and adjacent plateau areas of Iran. Each chromosome race had a discrete distribution, and the hexaploids were the most widespread. The diploids were cytologically regular, except for a chromosome interchange that occurred in about half of the plants. The tetraploids and hexaploids behaved cytologically as autopolyploids. The hexaploids were taller, coarser and later-flowering than the diploids and tetraploids, and they had fewer but thicker culms and larger seeds. The tetraploids were the leafiest and most productive, making them the most desirable from an agronomic standpoint. All races were more or less self-sterile, a characteristic that sets H. violaceum apart from most other Hordeum species. The taxonomic status of H. violaceum and its closest relatives, H. turkestanicum Nevski and H. brevisubulatum Link, is uncertain because of close morphological similarities and the occurrence of chromosome races in each taxon.     

国产驴蹄草的细胞地理学研究

为探讨国产毛茛科(Ranunculaceae)驴蹄草属(Caltha)两种植物的演化,该文利用传统染色体压片技术和流式细胞术,并结合前人染色体研究结果,对我国驴蹄草23个居群和花葶驴蹄草10个居群进行了细胞学研究。结果表明:驴蹄草是由四倍体(2n=4x=32)、六倍体(2n=6x=48)和八倍体(2n=8x=64)构成的多倍体复合群,花葶驴蹄草具有四倍体(2n=4x=32)和八倍体(2n=8x=64)两种倍性水平。驴蹄草和花葶驴蹄草均是四倍体较为常见,目前尚未见有二倍体报道。由于驴蹄草和花葶驴蹄草大部分居群采自中国青藏高原地区,可能在冰期时存在古二倍体,其适应性较弱,逐渐被其他的倍性取代,这是由于不同细胞型对环境适应性的结果。驴蹄草可能存在两条进化路线:一条是从甘肃到达云南;另一条是从西藏到达云南。前期分子系统学研究显示花葶驴蹄草与驴蹄草的亲缘关系较近,该研究结果中花葶驴蹄草染色体比驴蹄草要小,花葶驴蹄草可能比驴蹄草相对进化。目前花葶驴蹄草只有10个居群,还需进一步增加居群量来解析其演化路线。     

Relationships among genetic distance,forage yield and heterozygosity in isogenic diploid and tetraploid alfalfa populations

Isogenic diploid and tetraploid alfalfa (Medicago sativa L.) was studied with molecular markers to help understand why diploid performance and breeding behavior does not always predict that of tetraploids. In a previous study of partially heterozygous alfalfa genotypes, we detected a low correlation between yields of isogenic diploid (2x) and tetraploid (4x) single-cross progenies, and genetic distances were more highly correlated with yields of tetraploids than diploids. These differences may be related to the level of RFLP heterozygosity expected among progenies derived from heterozygous parents at the two ploidy levels. The objectives of this study were to determine the relationships among genetic distance, forage yield and heterozygosity in isogenic 2 x and 4 x alfalfa populations. Four diploid genotypes were chromosome doubled to produce corresponding isogenic autotetraploids, and these genotypes were mated in 4 × 4 diallels to produce 6 single-cross families at each ploidy level for field evaluation. Allele compositions of parents were determined at 33 RFLP loci by monitoring segregation of homologous restriction fragments among individuals within progenies, and these were used to estimate RFLP heterozygosity levels for all single-cross progenies at both ploidy levels. RFLP heterozygosity rankings were identical between progenies of isogenic diploid and tetraploid parents; but significant associations (P < 0.05) between estimated heterozygosity levels and forage yield were detected only at the tetraploid level. Since tetraploid families were nearly 25% more heterozygous than the corresponding diploid families, inconsistencies in the association between molecular marker diversity and forage yields of isogenic 2 x and 4 x single crosses may be due to recessive alleles that are expressed in diploids but masked in tetraploids. The gene action involved in heterosis may be the same at both ploidy levels; however, tetraploids benefit from greater complementary gene interactions than are possible for equivalent diploids. Present address: AgResearch Grasslands, New Zealand Pastoral Agriculture Research Institute, Palmerston North, New Zealand     

Morphological and environmental differentiation as prezygotic reproductive barriers between parapatric and allopatric Campanula rotundifolia agg. cytotypes

Background and AimsReproductive isolation and local establishment are necessary for plant speciation. Polyploidy, the possession of more than two complete chromosome sets, creates a strong postzygotic reproductive barrier between diploid and tetraploid cytotypes. However, this barrier weakens between polyploids (e.g. tetraploids and hexaploids). Reproductive isolation may be enhanced by cytotype morphological and environmental differentiation. Moreover, morphological adaptations to local conditions contribute to plant establishment. However, the relative contributions of ploidy level and the environment to morphology have generally been neglected. Thus, the extent of morphological variation driven by ploidy level and the environment was modelled for diploid, tetraploid and hexaploid cytotypes of Campanula rotundifolia agg. Cytotype distribution was updated, and morphological and environmental differentiation was tested in the presence and absence of natural contact zones.MethodsCytotype distribution was assessed from 231 localities in Central Europe, including 48 localities with known chromosome counts, using flow cytometry. Differentiation in environmental niche and morphology was tested for cytotype pairs using discriminant analyses. A structural equation model was used to explore the synergies between cytotype, environment and morphology.Key ResultsTremendous discrepancies were revealed between the reported and detected cytotype distribution. Neither mixed-ploidy populations nor interploidy hybrids were detected in the contact zones. Diploids had the broadest environmental niche, while hexaploids had the smallest and specialized niche. Hexaploids and spatially isolated cytotype pairs differed morphologically, including allopatric tetraploids. While leaf and shoot morphology were influenced by environmental conditions and polyploidy, flower morphology depended exclusively on the cytotype.ConclusionsReproductive isolation mechanisms vary between cytotypes. While diploids and polyploids are isolated postzygotically, the environmental niche shift is essential between higher polyploids. The impact of polyploidy and the environment on plant morphology implies the adaptive potential of polyploids, while the exclusive relationship between flower morphology and cytotype highlights the role of polyploidy in reproductive isolation.