Triploid Apomicts Research Articles

The Effect of Benzyl Amino Purin and NaphtalenaAcetic Acid Applications on Direct Shoot Organogenesis in Porang (Amorphophallus muelleri B)

Porang (Amorphophallus muelleri B.) is a tuberous plant with the potential to be the main source of carbohydrates and is rich in benefits. Porang proliferation is limited by a 6-month dormancy period per year, and generative propagation is unlikely due to the seeds being apomictic triploid. The aimed of my research to analyze the application of BAP and NAA in culture media for direct propagation of porang shoots. The explant used in this research was young leaves. The research was arranged in a completely randomized design with a combination of BAP and NAA hormones added to the MS medium. There were three BAP treatments, namely 1.0 mg/L, 2.0 mg/L, and 3.0 mg/L, while NAA treatments consisted of 2 levels, namely 2.0 mg/L and 4.0 mg/L. The addition of 1.0 mg/L BAP combined with 4.0 mg/L NAA was the best treatment that produced seven shoots with an average shoot length of 2.14 cm and root length of 3.6 cm, with the earliest bud emergence (9.7 weeks after planting).

Population structure, seasonal genotypic differentiation, and clonal diversity of weedy dandelions in three Boston area populations (Taraxacum sp.).

Weedy dandelions have a worldwide distribution and thrive in urban environments despite a lack of sexual reproduction throughout most of its range. North American dandelions, introduced from Eurasia, are believed to be primarily, if not exclusively, apomictic triploids. In some European populations, apomicts co‐occur with diploid sexual individuals and hybridizations can create genetically unique apomicts, which may subsequently disperse and establish new populations globally. Using six nuclear microsatellite markers and a cpDNA intergenic spacer, we investigate the impact of this unusual natural history on population structure and diversity in three urban Boston area dandelion populations. Our results show high levels of genetic diversity within populations, spatial population structure, and seasonal genotypic differentiation in flowering times. We find evidence that sexual reproduction and recombination, presumably in Europe, and extensive gene flow drive these patterns of diversity and create the appearance of panmixia despite the lack of evidence for local sexual reproduction.

Adapted dandelions trade dispersal for germination upon root herbivore attack.

A plant's offspring may escape unfavourable local conditions through seed dispersal. Whether plants use this strategy to escape insect herbivores is not well understood. Here, we explore how different dandelion (Taraxacum officinale agg.) populations, including diploid outcrossers and triploid apomicts, modify seed dispersal in response to root herbivore attack by their main root-feeding natural enemy, the larvae of the common cockchafer Melolontha melolontha. In a manipulative field experiment, root herbivore attack increased seed dispersal potential through a reduction in seed weight in populations that evolved under high root herbivore pressure, but not in populations that evolved under low pressure. This increase in dispersal potential was independent of plant cytotype, but associated with a reduction in germination rate, suggesting that adapted dandelions trade dispersal for establishment upon attack by root herbivores. Analysis of vegetative growth parameters suggested that the increased dispersal capacity was not the result of stress flowering. In summary, these results suggest that root herbivory selects for an induced increase in dispersal ability in response to herbivore attack. Induced seed dispersal may be a strategy that allows adapted plants to escape from herbivores.

Can obligate apomixis and more stable reproductive assurance explain the distributional successes of asexual triploids in Hieracium alpinum (Asteraceae)?

Although reproductive assurance has been suggested to be one of the most important factors shaping the differential distributional patterns between sexuals and asexuals (geographic parthenogenesis), it has only rarely been studied in natural populations of vascular plants with autonomous apomixis. Moreover, there are almost no data concerning the putative relationship between the level of apomictic versus sexual plant reproduction on one hand, and reproductive assurance on the other. We assessed the level of sexual versus apomictic reproduction in diploid and triploid plants of Hieracium alpinum across its distributional range using flow cytometric analyses of seeds, and compared the level of potential and realized seed set, i.e. reproductive assurance, between the two cytotypes under field and greenhouse conditions. Flow cytometric screening of embryos and endosperms of more than 4,100 seeds showed that diploids produced solely diploid progeny sexually, while triploids produced triploid progeny by obligate apomixis. Potential fruit set was much the same in diploids and triploids from the field and the greenhouse experiment. While in the pollination-limited environment in the greenhouse apomictic triploids had considerably higher realized fruit set than sexual diploids, there was no significant difference between cytotypes under natural conditions. In addition, sexuals varied to a significantly larger extent in realized fruit set than asexuals under both natural and greenhouse conditions. Our results indicate that triploid plants reproduce by obligate apomixis, assuring more stable and predictable fruit reproduction when compared to sexual diploids. This advantage could provide apomictic triploids with a superior colonisation ability, mirrored in a strong geographic parthenogenesis pattern observed in this species.

Complete chloroplast genomes from apomictic Taraxacum (Asteraceae): Identity and variation between three microspecies.

Chloroplast DNA sequences show substantial variation between higher plant species, and less variation within species, so are typically excellent markers to investigate evolutionary, population and genetic relationships and phylogenies. We sequenced the plastomes of Taraxacum obtusifrons Markl. (O978); T. stridulum Trávniček ined. (S3); and T. amplum Markl. (A978), three apomictic triploid (2n = 3x = 24) dandelions from the T. officinale agg. We aimed to characterize the variation in plastomes, define relationships and correlations with the apomictic microspecies status, and refine placement of the microspecies in the evolutionary or phylogenetic context of the Asteraceae. The chloroplast genomes of accessions O978 and S3 were identical and 151,322 bp long (where the nuclear genes are known to show variation), while A978 was 151,349 bp long. All three genomes contained 135 unique genes, with an additional copy of the trnF-GGA gene in the LSC region and 20 duplicated genes in the IR region, along with short repeats, the typical major Inverted Repeats (IR1 and IR2, 24,431bp long), and Large and Small Single Copy regions (LSC 83,889bp and SSC 18,571bp in O978). Between the two Taraxacum plastomes types, we identified 28 SNPs. The distribution of polymorphisms suggests some parts of the Taraxacum plastome are evolving at a slower rate. There was a hemi-nested inversion in the LSC region that is common to Asteraceae, and an SSC inversion from ndhF to rps15 found only in some Asteraceae lineages. A comparative repeat analysis showed variation between Taraxacum and the phylogenetically close genus Lactuca, with many more direct repeats of 40bp or more in Lactuca (1% larger plastome than Taraxacum). When individual genes and non-coding regions were for Asteraceae phylogeny reconstruction, not all showed the same evolutionary scenario suggesting care is needed for interpretation of relationships if a limited number of markers are used. Studying genotypic diversity in plastomes is important to characterize the nature of evolutionary processes in nuclear and cytoplasmic genomes with the different selection pressures, population structures and breeding systems.

Sexy males and sexless females: the origin of triploid apomicts.

Apomixis and polyploidy are closely associated in angiosperms, but the evolutionary reason for this association is unknown. Taraxacum officinale, the common dandelion, exists both as diploid sexuals and triploid apomicts. Here, in the context of T. officinale, we provide a model of the evolution of triploid apomicts from diploid sexuals. We posit an apomictic allele that arrests female meiosis in diploids, so that the plant produces diploid egg cells that can develop without fertilization, but haploid pollen. We propose occasional fertilization of diploid egg cells by haploid pollen, resulting in triploid apomicts that produce triploid egg cells but largely nonfunctional pollen. The irreversibility of this process renders diploid partial apomicts evolutionarily short-lived, and results in fixation of triploid apomicts except when they suffer extreme selective disadvantages. Our model can account for the high genetic diversity found in T. officinale triploid populations, because recombinant haploid pollen produced by diploids allows the apomictic allele to spread onto many genetic backgrounds. This leads to multiple clonal lineages in the newly apomictic population, and thereby alleviates some of the usual pitfalls of asexual reproduction.

Recent and dynamic transposable elements contribute to genomic divergence under asexuality.

BackgroundTransposable elements (TEs) are mobile pieces of genetic information with high mutagenic potential for the host genome. Transposition is often neutral or deleterious but may also generate potentially adaptive genetic variation. This additional source of variation could be especially relevant in non-recombining species reproducing asexually. However, evidence is lacking to determine the relevance of TEs in plant asexual genome evolution and their associated effects. Here, we characterize the repetitive fraction of the genome of the common dandelion, Taraxacum officinale and compare it between five accessions from the same apomictic lineage. The main objective of this study is to evaluate the extent of within-lineage divergence attributed to TE content and activity. We examined the repetitive genomic contribution, diversity, transcription and methylation changes to characterize accession-specific TEs.ResultsUsing low-coverage genomic sequencing, we report a highly heterogeneous TE compartment in the triploid apomict T. officinale representing up to 38.6 % of the homoploid genome. The repetitive compartment is dominated by LTR retrotransposon families accompanied by few non-LTR retrotransposons and DNA transposons. Up to half of the repeat clusters are biased towards very high read identity, indicating recent and potentially ongoing activity of these TE families. Interestingly, the five accessions are divided into two main clades based on their TE composition. Clade 2 is more dynamic than clade 1 with higher abundance of Gypsy Chromovirus sequences and transposons. Furthermore, a few low-abundant genomic TE clusters exhibit high level of transcription in two of the accessions analysed. Using reduced representation bisulfite sequencing, we detected 18.9 % of loci differentially methylated, of which 25.4 and 40.7 % are annotated as TEs or functional genes, respectively. Additionally, we show clear evidence for accession-specific TE families that are differentially transcribed and differentially methylated within the apomictic lineage, including one Copia Ale II LTR element and a PIF-Harbinger DNA transposon.ConclusionWe report here a very young and dynamic repetitive compartment that enhances divergence within one asexual lineage of T. officinale. We speculate that accession-specific TE families that are both transcriptionally and epigenetically variable are more prone to trigger changes in expression on nearby coding sequences. These findings emphasize the potential of TE-induced mutations on functional genes during asexual genome evolution.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3234-9) contains supplementary material, which is available to authorized users.

Depletion of Key Meiotic Genes and Transcriptome-Wide Abiotic Stress Reprogramming Mark Early Preparatory Events Ahead of Apomeiotic Transition.

Molecular dissection of apomixis – an asexual reproductive mode – is anticipated to solve the enigma of loss of meiotic sex, and to help fixing elite agronomic traits. The Brassicaceae genus Boechera comprises of both sexual and apomictic species, permitting comparative analyses of meiotic circumvention (apomeiosis) and parthenogenesis. Whereas previous studies reported local transcriptome changes during these events, it remained unclear whether global changes associated with hybridization, polyploidy and environmental adaptation that arose during evolution of Boechera might serve as (epi)genetic regulators of early development prior apomictic initiation. To identify these signatures during vegetative stages, we compared seedling RNA-seq transcriptomes of an obligate triploid apomict and a diploid sexual, both isolated from a drought-prone habitat. Uncovered were several genes differentially expressed between sexual and apomictic seedlings, including homologs of meiotic genes ASYNAPTIC 1 (ASY1) and MULTIPOLAR SPINDLE 1 (MPS1) that were down-regulated in apomicts. An intriguing class of apomict-specific deregulated genes included several NAC transcription factors, homologs of which are known to be transcriptionally reprogrammed during abiotic stress in other plants. Deregulation of both meiotic and stress-response genes during seedling stages might possibly be important in preparation for meiotic circumvention, as similar transcriptional alteration was discernible in apomeiotic floral buds too. Furthermore, we noted that the apomict showed better tolerance to osmotic stress in vitro than the sexual, in conjunction with significant upregulation of a subset of NAC genes. In support of the current model that DNA methylation epigenetically regulates stress, ploidy, hybridization and apomixis, we noted that ASY1, MPS1 and NAC019 homologs were deregulated in Boechera seedlings upon DNA demethylation, and ASY1 in particular seems to be repressed by global DNA methylation exclusively in the apomicts. Variability in stress and transcriptional response in a diploid apomict, which is geographically distinct from the triploid apomict, pinpoints both common and independent features of apomixis evolution. Our study provides a molecular frame-work to investigate how the adaptive traits associated with the evolutionary history of apomicts co-adapted with meiotic gene deregulation at early developmental stage, in order to predate meiotic recombination, which otherwise is thought to be favorable in stress and low-fitness conditions.

Estimating paternal efficiency in an agamic polyploid complex: pollen stainability and variation in pollen size related to reproduction mode, ploidy level and hybridogenous origin in Pilosella (Asteraceae)

Pollen quality was evaluated in Pilosella plants sourced from the field and representing various cytotypes (diploid to octoploid), reproduction modes (sexuality, facultative apomixis and seed sterility or semisterility) and status of origin (basic species, hybridogenous intermediate species and recent hybrids). Two methods of non-vital pollen staining were compared, providing a similar pattern of variation in pollen stainability and pollen size. A complex influence of ploidy level, reproduction mode and plant origin on pollen quality was found. Sexual biotypes had high and equable pollen stainability whereas apomictic and (semi)sterile plants were more variable in this trait. Nevertheless, the sexual, apomictic and (semi)sterile plants did not significantly differ in pollen stainability, while the impacts of both ploidy level and plant origin were significant. Apomictic triploids had low pollen stainability and heterogeneously sized pollen, most likely resulting from disturbed meiosis. The other biotypes, including higher odd-ploid cytotypes, displayed various patterns of pollen quality partially depending on the hybridogenous origin. Whereas high pollen quality was detected in most of the apomictic hexaploids and heptaploids, including recent hybrids, serious pollen dysfunction was found in most octoploids. Seed (semi)sterility was not strongly associated with reduced pollen quality. Either blocked or unstable pollen production was occasionally recorded, predominantly among recent hybrids. Despite autonomous apomixis independent of pollen, most apomictic biotypes displayed fairly high pollen stainability and pollen size homogeneity, implying sufficient production of viable reduced pollen. Most representatives of this agamic polyploid complex, including facultative apomicts, might efficiently mate as both male and female parents in natural populations.

Hybridization in natural mixed populations of sexual diploid and apomictic triploid dandelions (Taraxacum sect. Taraxacum): Why are the diploid sexuals not forced out?

In agamic complexes, the ‘minority cytotype exclusion principle’ can be modified to the ‘sexual cytotype exclusion principle’: Apomicts produce polyploid progeny of maternal genotype without the need for pollination, but still produce pollen. Outcrossing sexuals thus suffer from unsuccessful hybridizations – they produce reduced amounts of progeny. This causes a decrease in the relative frequency of sexuals and later leads to their exclusion from the population. However, in the agamic complex Taraxacum sect. Taraxacum, diploid sexuals coexist with triploid apomicts in stabilized mixed populations. This paper deals with hybridizations in natural populations, the possibilities of new triploid apomicts originating and investigates why the 'sexual cytotype exclusion principle' does not work in Taraxacum sect. Taraxacum. The progeny of diploids from natural populations screened by the flow-cytometric seed screening method consisted only of diploids. Different simulations of natural pollinations were carried experimentally. When the flowers of a diploid received different mixtures of pollen of surrounding diploids and triploids, pollination was successful and only diploid progeny was produced, probably due to the preference of high-quality compatible pollen of diploids by diploid maternal plants. Therefore, the 'sexual cytotype exclusion principle' cannot work in Taraxacum sect. Taraxacum, and diploids and triploids can coexist in common populations. Polyploid progeny was produced by a diploid plant surrounded only by triploid apomicts to a certain distance. This suggests that new polyploid apomicts can be produced only in places where isolated diploids can occur among triploids. Newly formed apomicts further spread their progeny to both areas of apomicts and areas of mixed populations.

Differential effects of polyploidy and diploidy on fitness of apomictic Boechera

The co-occurrence of apomixis (asexual reproduction) and polyploidy in plants has been the subject of debate in regard to the origin and evolution of asexuality. In recent years, polyploidy has been postulated as a maintenance and stabilization factor rather than as a source of apomixis origin. It is assumed polyploidy facilitates the compensation for mutation accumulation, and hence, the rare occurrence of diploid apomixis indirectly supports this finding. Nevertheless, diploid apomicts exist and are successful, especially in the genus Boechera. While comparing phenotypic traits, fitness-related traits and apomixis penetrance between both diploid and triploid apomicts in the genus Boechera, it was expected to find trait variance that can be attributed to ploidy. Surprisingly, little trait variation could be assigned to ploidy, but rather trait variations were mainly genotype-specific. Additionally, it is shown that paternal contribution is very important for trait success, even though all offspring are genetically identical to the mother plant. This harbors implications for the introduction of apomixis into crop plants, considering the effects of paternal contribution during asexual reproduction. Nevertheless, polyploidy is an efficient way to buffer deleterious mutations, but the flexibility of diploid apomicts of the genus Boechera for rare sexual events contributes to their success in nature.

Molecular and cytogenetic characterization of a collection of bahiagrass (Paspalum notatum Flügge) native to Uruguay

Paspalum notatum is a subtropical grass present throughout America, and one of the main constituents of the natural grasslands in Uruguay. An apomictic autotetraploid (2n = 4x = 40) is the most frequent cytotype. The occurrence of sexual diploids (2n = 2x = 20) has also been reported as well as the occasional presence of apomictic triploids and pentaploids in Argentina. In this study, ISSR (inter simple sequence repeats) molecular markers were used to analyze the genetic variability of 210 P. notatum individuals from a collection from Uruguay. Cytometric analyses and chromosome counts were used to assess the ploidy level of the individuals. All plants from Uruguay analyzed were tetraploid. Intra- and inter-population variability was found both in genomic DNA content and at the genotypic level. Several multilocus genotypes were shared among individuals within populations and among populations over moderate geographical ranges, at the same time, very dissimilar genotypes were found within the same population. Part of the genetic variance among populations can be explained by a broader scale geographic structure which is partly coincident with the traditionally recognized grassland management regions. In spite of the apparently high degree of genetic admixture within populations, groups of related genotypes seem to follow a broader geographical structure in the area under study. These results suggest that an efficient collection strategy for this apomictically reproducing species should include carefully planned intra- and inter-population sampling. A broader scale regional sampling strategy should also be considered although further studies will be required to define genetic structure at this level.

Modes of inheritance of two apomixis components, diplospory and parthenogenesis, in Chinese chive (<i>Allium ramosum</i>) revealed by analysis of the segregating population generated by back-crossing between amphimictic and apomictic diploids

To investigate the mode of inheritance of apomixis in Chinese chive, the degrees of diplospory and parthenogenesis were evaluated in F1 and BC1 progenies derived from crosses between amphimictic and apomictic diploids (2n = 16, 2x). The F1 population was generated by crossing three amphimictic diploids 94Mo13, 94Mo49 and 94Mo50 with an apomictic diploid KaD2 and comprised 110 diploids and 773 triploids. All the diploid F1 plants examined were completely or highly eusporous and completely syngamic. All the triploid F1 plants examined were highly diplosporous and highly parthenogenetic. KaD2 could not transmit its high level of apomixis via monoploid pollen grains. The BC1 population, generated by crossing 94Mo49 with apomictic triploids found in the F1 offspring, exhibited heteroploidy; it comprised haploid, diploid, triploid, tetraploid and various aneuploid individuals. In this generation, clear segregation was observed between diplospory and parthenogenesis. Analysis of the BC1 population suggests that diplospory and parthenogenesis are each controlled by single dominant genes, D and P, respectively. However, all the BC1 plants characterized as parthenogenetic were diplosporous. The absence of phenotypically eusporous parthenogenetic plants can be explained by assuming that the presence of diplospory gene is a prerequisite for the parthenogenesis gene expression in Chinese chive.

Is polyploidy a persevering accident or an adaptive evolutionary pattern? The case of the brine shrimp Artemia

Asexual organisms are confronted with substantial drawbacks, both immediate and delayed, threatening their evolutionary persistence. Yet, genetic associations with asexuality may refresh the gene pool promoting adaptation of clonal lineages; polyploidy is one of them. Parthenogenesis itself and/or polyploidy are responsible for the maintenance and spread of clones in Artemia, a sexual–asexual genus of halophilic anostracans. We applied flow cytometry, microsatellite genotyping, and mtDNA sequencing to 23 asexual populations. Artemia parthenogens have evolved multiple times either through hybridization or spontaneously. Nine out of 23 populations contained clones of mixed ploidy (2n, 3n, 4n). Most clones were diploid (20/31) while two and nine clones were triploid and tetraploid, respectively. Apomictic triploids and tetraploids formed two distinct groups of low genetic diversity compared with the more divergent automictic diploids. Polyploidy is also polyphyletic in Artemia, with triploids and tetraploids having independent origins from different sexual ancestors. We discern a pattern of geographical parthenogenesis with all clonal groups being more widespread than their closest sexuals. In favour of a specialist model, asexual diploids are restricted to single locations and are strikingly segregated from generalist triploids and tetraploids occupying a variety of sites. This is a rare pattern of mixed life-history strategies within an asexual complex.

Genetic homeostasis and developmental stability in natural populations of bisexual (Carassius auratus) and unisexual (C. gibelio) goldfishes

One amphymictic diploid Carassius auratus, three apomictic triploids C. gibbelio, and a hybrid triploid Carassius auratus forms of goldfish were compared with respect to a set of characteristics of developmental stability: morphological variation, fluctuating asymmetry, and phenodeviations. It was found that C. gibbelio forms are characterized by lower levels both of morphological variation and fluctuating symmetry and more rare morphological abnormalties. A hybrid form Carassius auratus-gibelio was characterized by the intermediate values of the above-listed features. It is emphasized that phenotypic stabilization of apomictic forms is caused by two factors: its clone structure and actual developmental canalization. It is specified that promoted genetic homeostasis of the C. gibelio form does not give obvious adaptive advantages, as in the basic river basins of East and Central Europe. they are everywhere ousted by an amphimictic C. auratus.

Deciphering the origins of apomictic polyploids in the Cheilanthes yavapensis complex (Pteridaceae)

Deciphering species relationships and hybrid origins in polyploid agamic species complexes is notoriously difficult. In this study of cheilanthoid ferns, we demonstrate increased resolving power for clarifying the origins of polyploid lineages by integrating evidence from a diverse selection of biosystematic methods. The prevalence of polyploidy, hybridization, and apomixis in ferns suggests that these processes play a significant role in their evolution and diversification. Using a combination of systematic approaches, we investigated the origins of apomictic polyploids belonging to the Cheilanthes yavapensis complex. Spore studies allowed us to assess ploidy levels; plastid and nuclear DNA sequencing revealed evolutionary relationships and confirmed the putative progenitors (both maternal and paternal) of taxa of hybrid origin; enzyme electrophoretic evidence provided information on genome dosage in allopolyploids. We find here that the widespread apomictic triploid, Cheilanthes lindheimeri, is an autopolyploid derived from a rare, previously undetected sexual diploid. The apomictic triploid Cheilanthes wootonii is shown to be an interspecific hybrid between C. fendleri and C. lindheimeri, whereas the apomictic tetraploid C. yavapensis is comprised of two cryptic and geographically distinct lineages. We show that earlier morphology-based hypotheses of species relationships, while not altogether incorrect, only partially explain the complicated evolutionary history of these ferns.

Geographical parthenogenesis, genome size variation and pollen production in the arctic-alpine species Hieracium alpinum

Hieracium alpinum L. (Asteraceae) is an arctic-alpine species distributed throughout Europe with both diploid and triploid cytotypes. We determined the ploidy levels of plants from 23 populations from Austria, Bosnia and Herzegovina, Finland, Italy, Norway, Romania, Slovakia, Switzerland and Ukraine. Data showed a non-overlapping pattern of cytotype distribution: sexually reproducing diploids (2n = 2x = 18) occur solely in the Eastern and Southern Carpathians, while apomictic triploids (2n = 3x = 27) cover the rest of the range. Such clear-cut allopatry is rather rare in vascular plants with geographical parthenogenesis. Comparison of absolute genome size indicates genome downsizing (by on average 3.7%) of haploid DNA amount in triploids relative to diploids. Genome size further correlated with longitude and latitude in the Alps, with decreasing absolute DNA content from west to east, and from south to north. While previously published data indicated complete male sterility of triploid plants, we found that plants from the Alps and Bosnia and Herzegovina commonly produced some pollen, whereas populations from the Western Carpathians and Scandinavia seemed to be almost completely pollen sterile. Scenarios about the evolution of geographical parthenogenesis in H. alpinum are discussed.

Population genetic structure of diploid sexual and polyploid apomictic hawthorns (Crataegus; Rosaceae) in the Pacific Northwest

Polyploidy and gametophytic apomixis are two important and associated processes in plants. Many hawthorn species are polyploids and can reproduce both sexually and apomictically. However, the population genetic structure of these species is poorly understood. Crataegus douglasii is represented exclusively by self-compatible tetraploid pseudogamous apomicts across North America, whereas Crataegus suksdorfii found in the Pacific Northwest is known to include self-incompatible diploid sexuals as well as polyploid apomicts. We compare population structure and genetic variability in these two closely related taxa using microsatellite and chloroplast sequence markers. Using 13 microsatellite loci located on four linkage groups, 251 alleles were detected in 239 individuals sampled from 15 localities. Within-population multilocus genotypic variation and molecular diversity are greatest in diploid sexuals and lowest in triploid apomicts. Apart from the isolation of eastern North American populations of C. douglasii, there is little evidence of isolation by distance in this taxon. Genetic diversity in western populations of C. douglasii suggests that gene flow is frequent, and that colonization and establishment are often successful. In contrast, local populations of C. suksdorfii are more markedly differentiated. Gene flow appears to be limited primarily by distance in diploids and by apomixis and self-compatibility in polyploids. We infer that apomixis and reproductive barriers between cytotypes are factors that reduce the frequency of gene flow among populations, and may ultimately lead to allopatric speciation in C. suksdorfii. Our findings shed light on evolution in woody plants that show heterogeneous ploidy levels and reproductive systems.

Chromosome numbers of 13 species in the genus Dryopteris (Dryopteridaceae) from Yunnan, China

Chromosomes of 13 species in the genus Dryopteris Adanson (Dryopteridaceae) from Yunnan, China have been counted. The numbers in four species, D. alpestris Tagawa (2n=82), D. habaensis Ching (2n=82), D. lachoongensis (Bedd.) Nayar & Kaur (2n=82), and D. yungtzeensis Ching (2n=82), were reported for the first time. The triploid apomict D. rosthornii (Diels) C. Chr. (2n=123), the tetraploid apomict D. wallichiana (Spreng.) Hylander ('n'=164, 2n=164) and the diploid apomict D. yoroii Serizawa (2n=82) represent three new cytotypes. Our results support the establishment of the section Caespitosae S. G. Lu, and indicate a high percentage of the diploid apomict cytotype in Dryopteris from the Himalayan region of China. The phenomenon of the presence of different cytotypes in a same plant of fern species is discussed.

The role of tetraploids in the sexual-asexual cycle in dandelions (Taraxacum).

Apomictic plants often produce pollen that can function in crosses with related sexuals. Moreover, facultative apomicts can produce some sexual offspring. In dandelions, Taraxacum, a sexual-asexual cycle between diploid sexuals and triploid apomicts, has been described, based on experimental crosses and population genetic studies. Little is known about the actual hybridization processes in nature. We therefore studied the sexual-asexual cycle in a mixed dandelion population in the Netherlands. In this population, the frequencies of sexual diploids and triploids were 0.31 and 0.68, respectively. In addition, less than 1% tetraploids were detected. Diploids were strict sexuals, triploids were obligate apomicts, but tetraploids were most often only partly apomictic, lacking certain elements of apomixis. Tetraploid seed fertility in the field was significantly lower than that of apomictic triploids. Field-pollinated sexual diploids produced on average less than 2% polyploid offspring, implying that the effect of hybridization in the 2x-3x cycle in Taraxacum will be low. Until now, 2x-3x crosses were assumed to be the main pathway of new formation of triploid apomicts in the sexual-asexual cycle in Taraxacum. However, tetraploid pollen donors produced 28 times more triploid offspring in experimental crosses with diploid sexuals than triploid pollen donors. Rare tetraploids may therefore act as an important bridge in the formation of new triploid apomicts.