Reproductive Assurance Research Articles

Severe inbreeding depression is predicted by the "rare allele load" in Mimulus guttatus.

Most flowering plants are hermaphroditic and experience strong pressures to evolve self-pollination (automatic selection and reproductive assurance). Inbreeding depression (ID) can oppose selection for selfing, but it remains unclear if ID is typically strong enough to maintain outcrossing. To measure the full cost of sustained inbreeding on fitness, and its genomic basis, we planted highly homozygous, fully genome-sequenced inbred lines of yellow monkeyflower (Mimulus guttatus) in the field next to outbred plants from crosses between the same lines. The cost of full homozygosity is severe: 65% for survival and 86% for lifetime seed production. Accounting for the unmeasured effect of lethal and sterile mutations, we estimate that the average fitness of fully inbred genotypes is only 3-4% that of outbred competitors. The genome sequence data provide no indication of simple overdominance, but the number of rare alleles carried by a line, especially within rare allele clusters nonrandomly distributed across the genome, is a significant negative predictor of fitness measurements. These findings are consistent with a deleterious allele model for ID. High variance in rare allele load among lines and the genomic distribution of rare alleles both suggest that migration might be an important source of deleterious alleles to local populations.

Do floral traits and the selfing capacity of Mimulus guttatus plastically respond to experimental temperature changes?

Climate change can negatively impact plant-pollinator interactions, and reduce outcross pollination. For reproductive assurance, an increased capacity for autonomous selfing should benefit the persistence of plants under new temperature conditions. Plastic responses of the autonomous selfing capacity to climate change may occur indirectly due to changes in floral traits associated with this capacity. We tested whether the mixed mating plant Mimulus guttatus is capable of plastic changes in floral traits favoring autonomous selfing in response to temperature changes. In seven growth chambers, we grew M. guttatus originating from a large range of latitudes (from 37.89° N to 49.95° N) and thus home temperatures in North America, and experimentally assessed the (autonomous) selfing and outcrossing capacities of the plants. With an increase in the difference between the overall mean daytime and nighttime experimental test temperature and home temperature, flower length and width decreased. The plastic response in flower size suggests that plants may be more successful at autonomous selfing. However, we did not find direct evidence that M. guttatus responded to increased temperature by an increased autonomous selfing capacity. With an increase in temperature difference, the odds of seed production, number of seeds, and individual seed mass decreased. Our results indicate that global warming and the associated increase in extreme temperature events may be detrimental to the reproduction and thus persistence of some plants.

Bees may drive the reproduction of four sympatric cacti in a vanishing coastal mediterranean-type ecosystem.

BackgroundSympatric congeneric plants might share pollinators, or each species might avoid competition by evolving specialized traits that generate partitions in pollinator assemblages. In both cases, pollen limitation (a decrease in the quality and quantity of compatible reproductive pollen) can occur, driving the plant mating system to autogamy as a mechanism of reproductive assurance. We assessed the relationships between pollinator assemblages and mating systems in a group of sympatric congeneric plants. We attempted to answer the following questions: (i) How similar are pollinator assemblages among sympatric cactus species? (ii) Which mating systems do sympatric cactus species use?MethodsWe studied sympatric Eriosyce taxa that inhabit a threatened coastal strip in a mediterranean-type ecosystem in central Chile. We performed field observations on four taxa and characterized pollinators during the years 2016 and 2017. We estimated differences in the pollinator assemblages using the Bray–Curtis index. To elucidate the mating systems, we conducted hand-pollination experiments using three treatments: manual cross-pollination, automatic self-pollination, and control (unmanipulated individuals). We tested differences in seed production for statistical significance using Kruskal–Wallis analysis.ResultsEriosyce subgibbosa showed a distinctive pollinator assemblage among the sympatric species that we studied (similarity ranged from 0% to 8%); it was visited by small bees and was the only species that was visited by the giant hummingbird Patagona gigas. Pollinator assemblages were similar between E. chilensis (year 2016 = 4 species; 2017 = 8) and E. chilensis var. albidiflora (2016 = 7; 2017 = 4); however, those of E. curvispina var. mutabilis (2016 = 7; 2017 = 6) were less similar to those of the aforementioned species. E. curvispina var. mutabilis showed the highest interannual variation in its pollinator assemblage (18% similarity). Reproduction in E. subgibbosa largely depends on pollinators, although it showed some degree of autogamy. Autonomous pollination was unfeasible in E. chilensis, which depended on flower visitors for its reproductive success. Both E. chilensis var. albidiflora and E. curvispina var. mutabilis showed some degree of autogamy.DiscussionWe observed differences in pollinator assemblages between E. subgibbosa and the remaining Eriosyce taxa, which depend on hymenopterans for pollen transfer. Pollinator assemblages showed considerable interannual variation, especially those of E. subgibbosa (ornithophilous syndrome) and E. curvispina var. mutabilis (melitophilous syndrome). Autogamous reproduction in these taxa may act as a reproductive assurance mechanism when pollinator availability is unpredictable. Our study contributes to improving our understanding of the reproductive systems of ecological interactions between threatened species in a Chilean mediterranean-type ecosystem.

Induced cleistogamy: A strategy for reproductive assurance in Murdannia nudiflora (Commelinaceae)

Murdannia nudiflora (L.) Brenan is a day flower that is dependent on entomophilous pollination. Despite the lack of pollinator attractants and its short flower longevity, M. nudiflora shows high rates of fecundity, which lead to their rapid dispersal. In addition to monitoring the vegetative reproductive methods and the effect of select environmental parameters (atmospheric temperature, relative humidity, and precipitation rates) in M. nudiflora, we evaluated mating systems in this species via artificial crosses. This species has evolved to undergo autogamy to ensure reproductive success when pollinators are insufficient; cleistogamous flowers are produced for optimal pollinator activity when environmental conditions are unfavorable. The number of cleistogamous flowers produced was found to be positively correlated with humidity and precipitation rates, and negatively correlated with temperature. In addition, it was found that high humidity and precipitation always hinder insect pollinators. Therefore, the combination of vegetative reproduction, entomophily, and cleistogamous self-pollination helps this weedy species to propagate with maximum efficiency. Murdannia nudiflora employs pre-anthesis cleistogamy as a mating strategy for reproductive assurance while retaining mechanisms that promote out-crossing.

Fitness costs of delayed pollination in a mixed-mating plant.

To predict the evolutionary consequences of pollinator declines, we need to understand the evolution of delayed autonomous self-pollination, which is expected to evolve as a mechanism of reproductive assurance when cross-pollination becomes unreliable. This involves estimating the costs of increased levels of selfing as well as those associated with floral senescence. We studied the mechanisms and costs of delayed self-pollination in the mixed-mating vine Dalechampia scandens (Euphorbiaceae) by first assessing among-population variation in herkogamy and dichogamy, which together determine the rate and timing of autonomous self-pollination. We then tested whether floral longevity responds plastically to delayed pollination. Finally, we assessed the costs of delayed self-pollination in terms of seed number and size, explicitly separating inbreeding depression from effects of floral senescence. Herkogamy varied extensively, while variation in dichogamy was more limited. Unpollinated blossoms increased their longevity, but seed quantity and quality decreased with increasing delays in pollination, independently of inbreeding depression. In D. scandens, earlier autonomous selfing is facilitated by reduced herkogamy rather than reduced protogyny, providing reproductive assurance while maintaining the possibility for outcrossing events. Effective early autonomous self-pollination may evolve under reduced cross-pollination reliability in response to costs associated with floral senescence.

Reproductive transitions in plants and animals: selfing syndrome, sexual selection and speciation.

The evolution of predominant self-fertilisation frequently coincides with the evolution of a collection of phenotypes that comprise the 'selfing syndrome', in both plants and animals. Genomic features also display a selfing syndrome. Selfing syndrome traits often involve changes to male and female reproductive characters that were subject to sexual selection and sexual conflict in the obligatorily outcrossing ancestor, including the gametic phase for both plants and animals. Rapid evolution of reproductive traits, due to both relaxed selection and directional selection under the new status of predominant selfing, lays the genetic groundwork for reproductive isolation. Consequently, shifts in sexual selection pressures coupled to transitions to selfing provide a powerful paradigm for investigating the speciation process. Plant and animal studies, however, emphasise distinct selective forces influencing reproductive-mode transitions: genetic transmission advantage to selfing or reproductive assurance outweighing the costs of inbreeding depression vs the costs of males and meiosis. Here, I synthesise links between sexual selection, evolution of selfing and speciation, with particular focus on identifying commonalities and differences between plant and animal systems and pointing to areas warranting further synergy.

Evolution of reproductive traits and selfing syndrome in the sub-endemic Mediterranean genus Centaurium Hill (Gentianaceae)

AbstractIn flowering plants, the shift from outcrossing to selfing is associated with a set of correlated changes in morphological and reproductive features known as the ‘selfing syndrome’. Species of the sub-endemic Mediterranean genus Centaurium Hill (Gentianaceae) exhibit a wide array of flower traits related to pollination biology and different ploidy levels. We explored if the evolutionary transitions of seven flower traits and life cycle, typically associated with the selfing syndrome, are related to polyploidy, diversification patterns, divergence times and the geological and climatic history of the Mediterranean Basin. Using 26 species of Centaurium we reconstructed a phylogenetic tree, inferred the ancestral states of the selected traits, estimated their phylogenetic signal and tested the correlative evolution among them. We found a significant increase in diversification rates during the Quaternary. Anther length, flower size, herkogamy and polyploidy undergo rapid state transitions without phylogenetic signal that could be the result of adaptation for selfing. Changes in character states do not show evidence of correlative evolution among them, as would be predicted during the evolution of selfing syndrome. The evolution of reproductive traits in Centaurium has probably relied on a more diverse array of drivers than just reproductive assurance or polyploidy events.

Isolation and Lack of Potential Mates may Threaten an Endangered Arid-Zone Acacia.

Clonality may provide reproductive assurance for many threatened plants while limiting sexual reproductive success either through energetic tradeoffs or because clones are self-incompatible. Most stands of the Australian arid-zone plant Acacia carneorum, flower annually but low seed set and an absence of sexual recruitment now suggest that this species and other, important arid-zone ecosystem engineers may have low genotypic diversity. Indeed, our recent landscape-scale genetic study revealed that stands are typically monoclonal, with genets usually separated by kilometers. An inability to set sexually produced seed or a lack of genetically diverse mates may explain almost system-wide reproductive failure. Here, using microsatellite markers, we genotyped 100 seeds from a rare fruiting stand (Middle-Camp), together with all adult plants within it and its 4 neighboring stands (up to 5 km distant). As expected, all stands surveyed were monoclonal. However, the Middle-Camp seeds were generated sexually. Comparing seed genotypes with the single Middle-Camp genotype and those of genets from neighboring and other regional stands (n = 26), revealed that 73 seeds were sired by the Middle-Camp genet. Within these Middle-Camp seeds we detected 19 genotypes in proportions consistent with self-fertilization of that genet. For the remaining 27 seeds, comprising 8 different genotypes, paternity was assigned to the nearest neighboring stands Mallee and Mallee-West, approximately 1 km distant. Ironically, given this species' vast geographic range, a small number of stands with reproductively compatible near neighbors may provide the only sources of novel genotypes.

Observations of Simultaneous Sperm Release and Larval Planulation Suggest Reproductive Assurance in the Coral Pocillopora acuta

Despite being an extensively studied group of corals, the reproductive biology of the scleractinian genus Pocillopora remains a mystery. Pocillopora acuta has been proposed to exhibit a mixed reproductive mode, sexually producing gametes (sperm and eggs) and asexually brooding larvae simultaneously within a single colony. Here, we report observations of night-time spawning of sperm during the peak monthly larval release period. We offer a new hypothesis for the regulation of sexual and asexual reproduction in the species and posit that sexual reproduction may occur more often than previously suggested. However, the success of internal oocyte fertilization and subsequent zygote development is dependent on sperm making contact with a fertile colony. We hypothesize that asexual development of larvae occurs when sperm is absent, but more extensive genetic, genomic, and histological data are required to determine the pathway by which unfertilized oocytes may develop. We also propose that this mixed mode of reproduction is an adaptation to mating failure, common in sessile marine invertebrates. The reproductive assurance enjoyed by the species may therefore be the key to its ecological and evolutionary persistence.

Phylogenomic analysis reveals multiple evolutionary origins of selfing from outcrossing in a lineage of heterostylous plants.

Evolutionary transitions from outcrossing to selfing often occur in heterostylous plants. Selfing homostyles originate within distylous populations and frequently evolve to become reproductively isolated species. We investigated this process in 10 species of Primula section Obconicolisteri using phylogenomic approaches and inferred how often homostyly originated from distyly and its consequences for population genetic diversity and floral trait evolution. We estimated phylogenetic relationships and reconstructed character evolution using the whole plastome comprised of 76 protein-coding genes. To investigate mating patterns and genetic diversity we screened 15 microsatellite loci in 40 populations. We compared floral traits among distylous and homostylous populations to determine how phenotypically differentiated homostyles were from their distylous ancestors. Section Obconicolisteri was monophyletic and we estimated multiple independent transitions from distyly to homostyly. High selfing rates characterised homostylous populations and this was associated with reduced genetic diversity. Flower size and pollen production were reduced in homostylous populations, but pollen size was significantly larger in some homostyles than in distylous morphs. Repeated transitions to selfing in section Obconicolisteri are likely to have been fostered by the complex montane environments that species occupy. Unsatisfactory pollinator service is likely to have promoted reproductive assurance in homostyles leading to subsequent population divergence through isolation.

Water mediates fertilization in a terrestrial flowering plant.

Water-mediated fertilization is ubiquitous in early land plants. This ancestral mode of fertilization has, however, generally been considered to have been lost during the evolutionary history of terrestrial flowering plants. We investigated reproductive mechanisms in the subtropical ginger Cautleya gracilis (Zingiberaceae), which has two pollen conditions - granular and filiform masses - depending on external conditions. We tested whether rain transformed granular pollen into filiform masses and whether this then promoted pollen-tube growth and fertilization of ovules. Using experimental manipulations in the field we investigated the contribution of water-mediated fertilization to seed production. Rain caused granular pollen to form filiform masses of germinating pollen tubes, which transported sperm to ovules, resulting in fertilization and seed set. Flowers exposed to rain produced significantly more seeds than those protected from the rain, which retained granular pollen. Insect pollination made only a limited contribution to seed set because rainy conditions limited pollinator service. Our results reveal a previously undescribed fertilization mechanism in flowering plants involving water-mediated fertilization stimulated by rain. Water-mediated fertilization is likely to be adaptive in the subtropical monsoon environments in which C.gracilis occurs by ensuring reproductive assurance when persistent rain prevents insect-mediated pollination.

A functional decomposition of sex inconstancy in the dioecious, colonizing plant Mercurialis annua.

Plants with separate sexes often show "inconstant" or "leaky" sex expression, with females or males producing a few flowers of the opposite sex. The frequency and degree of such inconstancy may reflect residual hermaphroditic sex allocation after an evolutionary transition from combined to separate sexes. Sex inconstancy also represents a possible first step in the breakdown of dioecy back to hermaphroditism. In the Mercurialis annua (Euphorbiaceae) species complex, monoecy and androdioecy have evolved from dioecy in polyploid populations. Here, we characterize patterns of sex inconstancy in dioecious M. annua and discuss how sex inconstancy may have contributed to the breakdown of separate sexes in the genus. We measured sex inconstancy in three common gardens of M. annua over 2 years using a modification of Lloyd's phenotypic gender in terms of frequency and degree, with the degree calibrating inconstancy against the sex allocation of constant males and constant females, yielding a measure of gender that does not depend on the distribution of gender in the population. Unusually for dioecious plants, the frequency of sex inconstancy in M. annua was greater in females, but its degree was greater for males in the 2 years of study. We suggest that this pattern is consistent with the maintenance of inconstancy in dioecious M. annua by selection for reproductive assurance under mate limitation. Our study illustrates the utility of decomposing measures of sex inconstancy into its frequency and its degree and throws new light on the origin of variation in sexual systems in Mercurialis.

Geographic variation in reproductive assurance of Clarkia pulchella.

Climate can affect plant populations through direct effects on physiology and fitness, and through indirect effects on their relationships with pollinating mutualists. We therefore expect that geographic variation in climate might lead to variation in plant mating systems. Biogeographic processes, such as range expansion, can also contribute to geographic patterns in mating system traits. We manipulated pollinator access to plants in eight sites spanning the geographic range of Clarkia pulchella to investigate geographic and climatic drivers of fruit production and seed set in the absence of pollinators (reproductive assurance). We examined how reproductive assurance and fruit production varied with the position of sites within the range of the species and with temperature and precipitation. We found that reproductive assurance in C. pulchella was greatest in populations in the northern part of the species' range and was not well explained by any of the climate variables that we considered. In the absence of pollinators, some populations of C. pulchella have the capacity to increase fruit production, perhaps through resource reallocation, but this response is climate dependent. Pollinators are important for reproduction in this species, and recruitment is sensitive to seed input. The degree of autonomous self-pollination that is possible in populations of this mixed-mating species may be shaped by historic biogeographic processes or variation in plant and pollinator community composition rather than variation in climate.

Rapid loss of self-incompatibility in experimental populations of the perennial outcrossing plant Linaria cavanillesii.

Transitions from self-incompatibility to self-compatibility in angiosperms may be frequently driven by selection for reproductive assurance when mates or pollinators are rare, and are often succeeded by loss of inbreeding depression by purging. Here, we use experimental evolution to investigate the spread of self-compatibility from one such population of the perennial plant Linaria cavanillesii into self-incompatible (SI) populations that still have high inbreeding depression. We introduced self-compatible (SC) individuals at different frequencies into replicate experimental populations of L. cavanillesii that varied in access to pollinators. Our experiment revealed a rapid shift to self-compatibility in all replicates, driven by both greater seed set and greater outcross siring success of SC individuals. We discuss our results in the light of computer simulations that confirm the tendency of self-compatibility to spread into SI populations under the observed conditions. Our study illustrates the ease with which self-compatibility can spread among populations, a requisite for species-wide transitions from self-incompatibility to self-compatibility.

Reproductive assurance weakens pollinator-mediated selection on flower size in an annual mixed-mating species.

In animal-pollinated plants, direct and indirect selection for large and small flowers in predominantly outcrossing and selfing species, respectively, is a common consequence of pollen limitation (PL). However, many hermaphroditic species show a mixed-mating system known as delayed selfing, which provides reproductive assurance (RA) only when outcrossing is not realized. Although RA is expected to reduce pollinator-mediated selection towards larger flowers, the consequences of delayed selfing for selection on flower size in mixed-mating species remain overlooked. We investigated whether RA weakens selection on flower size in Tuberaria guttata, a mixed-mating annual herb. We related pollinator visitation rates to flower size and measured seed production in emasculated, hand cross-pollinated and intact (control) flowers in three natural populations. For each population, we estimated variation in PL and RA across individuals differing in flower size and phenotypic selection on this trait. Pollinator visitation increased and RA decreased with flower size in all populations. Increasing RA diminished but did not fully alleviate PL, because of early-acting inbreeding depression. In the least-visited and most pollen-limited population, RA increased seed production by >200 %, intensely counteracting the strong pollinator-mediated selection for larger corollas. In the most-visited population, however, RA increased seed production by an average of only 9 %. This population exhibited the largest fraction of individuals that showed a decrease in seed production due to selfing and the weakest pollinator-mediated selection on flower size. The results suggest that the balance between the extent of RA and outcrossing contributes to determine flower size in mixed-mating systems. Pollinator-mediated selection favours larger flowers by increasing outcrossed seeds, but the benefits of RA greatly lessen this effect, especially under severe conditions of pollen limitation. Our findings also indicate that a mixed-mating system can represent an 'evolutionary trap' under an adequate pollinator supply.

The Effect of Pollen Limitation on the Evolution of Mating System and Seed Size in Hermaphroditic Plants.

Pollen limitation, when inadequate pollen receipt results in a plant setting fewer seeds and fruits, can reduce plant reproductive success and promote the evolution of self-fertilization as a mechanism of reproductive assurance. However, the effect of pollen limitation on the joint evolution of mating system and seed size is not known. Using an evolutionarily stable strategy resource allocation model, we show that where moderate pollen limitation and strong inbreeding depression select for complete outcrossing, pollen limitation should also increase the optimal seed size. In contrast, pollen limitation should not affect the optimal seed size under complete selfing, in which case ovule fertilization is certain. Under intermediate conditions, a mixed mating system evolves if the probability of ovule fertilization declines as more ovules are produced, so that a selfed seed with inbreeding depression provides equal marginal fitness returns to a larger outcrossed seed that may result from pollen limitation. Under mixed mating, outcrossed seeds should be larger than selfed seeds, and pollen limitation should not affect the optimal size of either outcrossed or selfed seeds. Our results identify a novel pathway through which pollen limitation selects for mixed mating and provide an adaptive explanation as to why selfed seeds are often smaller than outcrossed seeds.

Floral Behaviour, Flowering Phenology and Fruit Production of Passion Fruit (Passiflora Species) in East Malaysia

Passiflora species generally known as passion fruit may well be the most fascinating plant of the tropics. This plant is known to be self-incompatible in its native environment and it affects the fruit production. Extend of passion fruit set is highly dependent on effective pollinators such as carpenter bees and honey bees. However, in East Malaysia, there was good fruit sets, although lesser number of pollinators visiting the farm or sometimes absence. It is important therefore, to study the reproduction mechanisms of Passiflora plants, because this factor reflect the quantity of the fruit produced. The present studies aim therefore to evaluate the floral behaviours, reproductive mechanism and fruit production of locally grown Passiflora species. Based on the observations, Passiflora species was classified as “steadystate species” that exhibit constant production of few flowers, each day lasting only one day. All species have the ability to adapt to the local condition where plants continuously grew and produced flowers and fruits all the year around. The flowering and fruiting were not synchronized and showed a marked phenological pattern with species. Good fruit yields were observed throughout the year which was attributed to Passiflora species ability for self-pollination, involving the movement of styles to lead the stigma faced the anthers during flower blooming. This is an important phenomenon in order to reduce the distance between the stigma and anther, aid for pollination and provided reproductive assurance to the species. As P. edulis growing in its native environment is reported to be self-incompatible and have become selfcompatible in the present study; showed that evolution of these features is an important factor for its adaptability and survivability, thus crucial to P. edulis propagation optimally in local conditions.

A Novel Autogamous Self-Pollination Strategy Involving Closing of Perianth Lobes in Talinum fruticosum (L.) Juss.

Pollination is a prerequisite for fertilization and subsequent fruit and seed development. Nearly 90% of the flowering plants use a range of animals for pollination services. Depending on the sexuality and the breeding system of the species, the type of pollen that the flower receives ranges from complete outcrossing to complete selfing. Preliminary observations on Talinum fruticosum (L.) Juss. (Portulacaceae) did not reveal any floral visitors but showed abundant fruit and seed set. Detailed studies carried out to analyze the mode of development of fruits and seeds in the absence of pollinators revealed a novel method of autogamy in this species. Diurnal flowers remain open just for 5–6 h, and they do not get pollinated during this period due to lack of floral visitors. The five free perianth lobes start closing up and gradually curl around the stamens and the pistil, thus bringing the dehisced anthers in contact with the stigma in almost all the flowers inducing > 90% fruit set. This is another example of the evolution of autogamy under pollinator constraint as a means of reproductive assurance.

Reproductive trade-offs maintain bract color polymorphism in Scarlet Indian paintbrush (Castilleja coccinea).

Populations of scarlet Indian paintbrush (Castilleja coccinea) in the Midwestern United States exhibit a bract color polymorphism, with each population having predominantly yellow or scarlet bracts. We investigated a possible mechanism for this maintenance of bract color polymorphism in C. coccinea by conducting hand-pollination experiments in two nearby populations, one predominantly yellow and one predominantly scarlet. The hand-pollination treatments were either self-pollination or cross pollination using pollen from within and between populations. Both color morphs were used as pollen donors for the within and between crosses. We found that both color morphs of C. coccinea were self-compatible. When the scarlet morph was the maternal plant it had higher seed set. When pollinators were excluded, the yellow morph outperformed the scarlet morph in fruit set and seed set. The apparent trade-offs between a higher reproductive output in the scarlet morph and a reproductive assurance advantage in the yellow morph may explain the maintenance of the polymorphism in C. coccinea. While many previous studies have provided evidence for pollinator preference playing a role in floral color polymorphism, the results of the current study indicate that reproductive assurance, which would be important for fluctuations in pollinator abundance or colonizing new areas, may act as a selective agent to maintain such polymorphisms.

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.