Post-pollination Barriers Research Articles

Reproductive isolation between two sympatric bat-pollinated Bauhinia (Leguminosae).

Several barriers contribute to reproductive isolation between plant species, which can be classified as pre- or post-pollination. Understanding the strength of these barriers could clarify the factors that maintain reproductive isolation and thus species integrity. In this study, we quantified reproductive isolation between two bat-pollinated co-occurring Bauhinia species (B. acuruana and B. pentandra) with similar flower morphology. Over the course of 18 months, we assessed reproductive isolation between these two Bauhinia species by quantifying the individual strengths and absolute contributions of five pre- and post- pollination barriers. Our data showed that both species are completely isolated in their reproduction by a combination of several barriers. Although they co-occur in a few populations, we found a high degree of geographic isolation between them. And although their flowering periods overlap, there is a significant difference in flowering peaks. Both species have the same pollinating bats, but the interspecific transfer of pollen between the plant species may be reduced due to the different length of the flower stamens, resulting in different pollen deposition on the bats' bodies. We have documented complete incompatibility between taxa and conclude that pre- and post-pollination barriers are important factors in preventing gene flow, even in contact zones between these two species of Bauhinia. We highlight that our work is the first study to use methods to estimate the strength of reproductive isolation barriers between bat-pollinated species.

Recent parallel speciation in Antirrhinum involved complex haplotypes and multiple adaptive characters.

A role of ecological adaptation in speciation can be obscured by stochastic processes and differences that species accumulate after genetic isolation. One way to identify adaptive characters and their underlying genes is to study cases of speciation involving parallel adaptations. Recently resolved phylogenies reveal that alpine morphology has evolved in parallel in the genus Antirrhinum (snapdragons): first in an early split of an alpine from a lowland lineage and, more recently, from within the lowland lineage to produce closely related sympatric species with contrasting alpine and lowland forms. Here, we find that two of these later diverged sympatric species are differentiated by only around 2% of nuclear loci. Though showing evidence of recent gene flow, the species remain distinct for a suite of morphological characters typical of earlier-diverged alpine or lowland lineages and their morphologies correlate with features of the local landscape, as expected of ecological adaptations. Morphological differences between the two species involve multiple, unlinked genes so that parental character combinations are readily broken up by recombination in hybrids. We detect little evidence for post-pollination barriers to gene flow or recombination, suggesting that genetic isolation related to ecological adaptation is important in maintaining character combinations and might have contributed to parallel speciation. We also find evidence that genes involved in the earlier alpine-lowland split were reused in parallel evolution of alpine species, consistent with introgressive hybridisation, and speculate that many non-ecological barriers to gene flow might have been purged during the process.

Mechanisms of prezygotic post-pollination reproductive barriers in plants.

Hybridisation between individuals of different species can lead to maladapted or inviable progeny due to genetic incompatibilities between diverging species. On the other hand, mating with close relatives, or self-fertilisation may lead to inbreeding depression. Thus, both too much or too little divergence may lead to problems and the organisms have to carefully choose mating partners to avoid both of these pitfalls. In plants this choice occurs at many stages during reproduction, but pollen-pistil interactions play a particularly important role in avoiding inbreeding and hybridisation with other species. Interestingly, the mechanisms involved in avoidance of selfing and interspecific hybridisation may work via shared molecular pathways, as self-incompatible species tend to be more 'choosy' with heterospecific pollen compared to self-compatible ones. This review discusses various prezygotic post-pollination barriers to interspecific hybridisation, with a focus on the mechanisms of pollen-pistil interactions and their role in the maintenance of species integrity.

Eco-Geography and Phenology Are the Major Drivers of Reproductive Isolation in the Royal Irises, a Species Complex in the Course of Speciation

The continuous nature of speciation implies that different species are found at different stages of divergence, from no- to complete reproductive isolation. This process and its underlying mechanisms are best viewed in incipient species. Moreover, the species complex can offer unique insight into how reproductive isolation (RI) has evolved. The royal irises (Iris section Oncocyclus) are a young group of species in the course of speciation, providing an ideal system for speciation study. We quantified pre- and post-zygotic reproductive barriers between the eight Israeli species of this complex and estimated the total RI among them. We tested for both pre-pollination and post-pollination reproductive barriers. Pre-pollination barriers, i.e., eco-geographic divergence and phenological differentiation were the major contributors to RI among the Iris species. On the other hand, post-pollination barriers, namely pollen-stigma interactions, fruit set, and seed viability had negligible contributions to total RI. The strength of RI was not uniform across the species complex, suggesting that species may have diverged at different rates. Overall, this study in a young, recently diverged group of species provides insight into the first steps of speciation, suggesting a crucial role of the pre-zygotic barriers.

Pre- and post-pollination barriers between two exotic and five native Sagittaria species: Implications for species conservation

Anthropogenic introduction of species has resulted in a breakdown of geographical barriers and hybridization in previously allopatric species. Thus, examining hybridization proneness of exotic species contributes to revealing its potential threat. Moreover, reproductive barriers may be strengthened or weakened due to long-term geographical isolation for these newly sympatric species. However, few studies have evaluated multiple barriers between alien and native species. In this study, we quantified the importance of four pre-pollination barriers (phenological, floral traits, pollen production, and floral constancy) and four post-pollination barriers (pollen-pistil incompatibility, seed set, seed viability, and seedling survival) between two introduced and five native Sagittaria species. Results showed that introduced S. platyphylla was cross-compatible with two native species, whereas introduced S. montevidensis was incapable of hybridizing with any native species. Different barriers were asymmetric within species pairs and multiple barriers acted in concert to maintain species boundaries. Post-pollination barriers contributed more to total reproductive isolation in native species, whereas pre-pollination barriers played a stronger role in total reproductive isolation for two introduced species. Seed set was the only barrier that was positively correlated with genetic distance. Our results provide a perspective to better understand reproductive barriers for secondary contact species. We highlight the importance of monitoring hybridization events before human introduction and the possible conservation strategies to remove invasive species with hybridization proneness.

Microhabitat and Pollinator Differentiation Drive Reproductive Isolation between Two Sympatric Salvia Species (Lamiaceae).

Evaluation of multiple barriers contributing to reproductive isolation between sympatric plant species is key to understanding the mechanism of their coexistence; however, such investigations in biodiversity hotspots are still rare. In this study, we investigated and compared geography, microhabitat, phenology, flora, and pollinators, in addition to pollen–pistil interactions, seed production, and seed germination of the closely related sympatric Salvia digitaloides and S. flava on Yulong Snow Mountain, Southwestern Yunnan, China. The geographic distribution of these species overlapped, but their adaptation to physical and chemical properties of soil microhabitats differed. They shared the same flowering time but differed in flower size, style length, nectar volume, sugar concentration, and flower longevity. Both species shared bumblebees as effective pollinators, but flower constancy for the two species was relatively strong. Pollen tube growth, seed production, and seed germination were lower in interspecific than in intraspecific crosses. Our study suggested that microhabitat and pollinator isolation acted as the most important isolating barriers in maintaining the coexistence of the two Salvia species. Our study also highlighted that post-pollination barriers play an important role in preventing the gene flow between these two Salvia species.

Reproductive Isolation Among Three Nocturnal Moth-Pollinated Sympatric Habenaria Species (Orchidaceae).

Comparison and quantification of multiple pre- and post-pollination barriers to interspecific hybridization are important to understand the factors promoting reproductive isolation. Such isolating factors have been studied recently in many flowering plant species which seek after the general roles and relative strengths of different pre- and post-pollination barriers. In this study, we quantified six isolating factors (ecogeographic isolation, phenological isolation, pollinator isolation, pollinia-pistil interactions, fruit production, and seed development) that could possibly be acting as reproductive barriers at different stages among three sympatric Habenaria species (H. limprichtii, H. davidii, and H. delavayi). These three species overlap geographically but occupy different microhabitats varying in soil water content. They were isolated through pollinator interactions both ethologically (pollinator preference) and mechanically (pollinia attachment site), but to a variable degree for different species pairs. Interspecific crosses between H. limprichtii and H. davidii result in high fruit set, and embryo development suggested weak post-pollination barriers, whereas bidirectional crosses of H. delavayi with either of the other two species fail to produce fruits. Our results revealed that pollinators were the most important isolating barrier including both ethological and mechanical mechanisms, to maintain the boundaries among these three sympatric Habenaria species. Our study also highlights the importance of a combination of pre-and post-pollination barriers for species co-existence in Orchidaceae.

Speciation with gene flow between two Neotropical sympatric species (Pitcairnia spp.: Bromeliaceae).

The study of mechanisms that generate new species is considered fundamental for broad areas of ecology and evolution. Speciation is a continuous process in which reproductive isolation is established, and it is of fundamental importance to understand the origins of the adaptations that contribute to this process. Hybrid zones are considered natural laboratories for the study of speciation and represent ideal systems for such studies. Here, we investigated genomic differentiation between hybridizing Neotropical species Pitcairnia staminea (G. Lodd.) and P. albiflos (Herb.). Using thousands of SNPs genotyped through RAD‐seq, we estimate effective population sizes, interspecific gene flow, as well as time of divergence between these two sister species and identify candidate genomic regions for positive selection that may be related to reproductive isolation. We selected different scenarios of speciation and tested them by using approximate Bayesian computation (ABC); we found evidence of divergence with gradual reduction in gene flow between these species over time, compatible with the hypothesis of speciation with gene flow between these Pitcairnia species. The parameter estimates obtained through ABC suggested that the effective population size of P. albiflos was around three times larger than that of P. staminea. Our divergence date estimates showed that these two species diverged during the Pliocene (4.7 Mya; CI = 1.3–8.5 Mya), which has likely allowed this species to accumulate genome‐wide differences. We also detected a total of 17 of 4165 loci which showed signatures of selection with high genetic differentiation (F ST > 0.85), 12 of these loci were annotated in de novo assembled transcriptomes of both species, and 4 candidate genes were identified to be putatively involved in reproductive isolation. These four candidate genes were previously associated with the function of pollen development, pollen tube germination and orientation, abiotic stress, and flower scent in plants, suggesting an interplay between pre‐ and postpollination barriers in the evolution of reproductive isolation between such species.

Floral morpho-anatomy and reproductive ecology of Spondias macrocarpa Engl. (Anacardiaceae), a vulnerable neotropical andromonoecious tree

The genus Spondias has a wide variety of sexual systems, with complete monoecious to polygamodioecious species. Spondias macrocarpa (Anacardiaceae) is an endemic species from Brazil, previously described as hermaphroditic, and considered vulnerable due to massive deforestation. However, to our knowledge, no further details on the breeding system or morphological studies are available. Herewith, we examine the floral morpho-anatomy and breeding system of this species to better understand the sexual function of different floral morphologies and the role of pollinators in its reproduction. We used light and scanning electron microscopy to study the morphology and anatomy of the flowers. Additionally, we carried out experiments on pollen viability, stigmatic receptivity, and controlled pollinations. We found both bisexual and male flowers in the same inflorescence, with the staminate flowers presenting cryptic unisexuality. Investment in bisexual flowers was higher than in male ones, although more than 70% of the inflorescence was made up of male flowers. Most male flowers are present in the proximal region of the inflorescence, and there is no temporal separation between male and female functions in bisexual flowers. We show that S. macrocarpa is andromonoecious with cryptic-monoecy and a self-incompatibility system. Under natural conditions, S. macrocarpa is probably a pollinator generalist, with small bees being its main floral visitors. High levels of pollen robbing and low levels of crossing by floral visitors may result in the observed low fruit set. Further studies on the pre- and post-pollination barriers and on the reproductive ecology of extant natural populations should elucidate the reason for such a low fruit set.

Post-pollination barriers enable coexistence of pollinator-sharing ornithophilous Erica species.

Some evolutionary radiations produce a number of closely-related species that continue to coexist. In such plant systems, when pre-pollination barriers are weak, relatively strong post-pollination reproductive barriers are required to maintain species boundaries. Even when post-pollination barriers are in place, however, reproductive interference and pollinator dependence may strengthen selection for pre-pollination barriers. We assessed whether coexistence of species from the unusually speciose Erica genus in the fynbos biome, South Africa, is enabled through pre-pollination or post-pollination barriers. We also tested for reproductive interference and pollinator dependence. We investigated this in natural populations of three bird-pollinated Erica species (Erica plukenetii, E. curviflora and E. coccinea), which form part of a large guild of congeneric species that co-flower and share a single pollinator species (Orange-breasted Sunbird Anthobaphes violacea). At least two of the three pre-pollination barriers assessed (distribution ranges, flowering phenology and flower morphology) were weak in each species pair. Hand-pollination experiments revealed that seed set from heterospecific pollination (average 8%) was significantly lower than seed set from outcross pollination (average 50%), supporting the hypothesis that species boundaries are maintained through post-pollination barriers. Reproductive interference, assessed in one population by applying outcross pollen three hours after applying heterospecific pollen, significantly reduced seed set compared to outcross pollen alone. This may drive selection for traits that enhance pre-pollination barriers, particularly given that two of the three species were self-sterile, and therefore pollinator dependent. This study suggests that post-pollination reproductive barriers could facilitate the coexistence of congeneric species, in a recent radiation with weak pre-pollination reproductive barriers.

Reproductive isolation between wild and domesticated chaya (Cnidoscolus aconitifolius) in sympatry.

Reproductive isolation is a necessary condition for plant domestication in their domestication centre where crops co-occur with their wild progenitors. However, the identification of reproductive barriers and their relative contribution to reproductive isolation have been overlooked in plants under domestication. We assessed pre- and post-pollination reproductive barriers and their relative contribution to reproductive isolation between wild and domesticated chaya (Cnidoscolus aconitifolius) in its domestication centre. We found that wild and domesticated chaya both exhibit a high degree of reproductive isolation. However, the reproductive isolation barriers exhibited some asymmetry: while pre-pollination barriers (differential pollen production and pollinator specificity) were only detected in wild plants, post-pollination barriers (pollen-pistil incompatibility and/or failure to set fruit) were observed in both wild and domesticated plants. We conclude that complete reproductive isolation has evolved in sympatry in co-occurring domesticated and wild chaya.

Pre-zygotic reproductive isolation between two synchronopatric Opuntia (Cactaceae) species in the Brazilian Chaco.

Opuntia (Cactaceae) is known for high rates of hybridization and ploidisation, resulting in the formation of new species. The occurrence of two sympatric and closely related species of Opuntia, O.elata and O.retrorsa, in Brazilian Chaco enabled us to test the hypothesis that pre-zygotic reproductive isolation mechanisms operate in both species. We monitored the flowering period, as well as floral biology, and compared the morphological variation of floral structures through measurements, performed intra- and interspecific cross-pollination tests, and recorded the guild of floral visitors and pollinators. Flowering was seasonal and highly synchronous. Floral biology exhibits similar strategies, and although floral morphology differs significantly in many of the compared structures, such morphological variation does not result in the selection of exclusive pollinators. Floral visitors and pollinators are oligolectic bees shared by both species. Opuntia elata and O.retrorsa are self-compatible. While interspecific cross-pollination (bidirectional) resulted in germination, the pollen tube did not penetrate the stigma. Opuntia elata and O.retrorsa are closely related; however, they are isolated and do not hybridise in Brazilian Chaco. We found that both have weak pre-pollination barriers, but that they are strongly isolated by pollen-pistil incompatibility, i.e. post-pollination barrier.

Reproductive systems and post-pollination barriers between two closely related eu-bromelioids (Bromeliaceae) in the Atlantic Forest of Brazil

Abstract The eu-bromelioid clade provides a classic example of an exceptionally rapid radiation, in which many closely related species are found in sympatry. In this respect, species cohesion may be due to the rapid evolution of strong reproductive barriers, although interspecific and intergeneric hybridization is observed in the family. The objective of this study was to characterize the breeding systems and post-pollination reproductive barriers in Wittmackia patentissima and Hohenbergia ridleyi. Previously included in Aechmea, they show an almost complete absence of pre-pollination barriers, although their recent divergence may not have allowed time for complete ecological/reproductive isolation to have evolved. In manual pollinations, both species had > 40% fruit set in self and outcross treatments, but seed set from manual self-pollinations was < 50% that of outcrosses, and spontaneous self-pollination produced fewer than three seeds per fruit. Self-pollen and pollen tubes showed no evidence of self-incompatibility, but heterospecific pollen tubes were arrested in the lower part of the style and heterospecific seed set was zero in both species. The combination of self-compatibility and strong post-pollination, pre-fertilization reproductive barriers is consistent with the involvement of breeding system reproductive barriers in allowing closely related bromeliad species to occur in sympatry.

Reproductive isolation among three sympatric Achimenes species: pre- and post-pollination components.

Closely related species occurring in sympatry may experience the negative consequences of interspecific pollen transfer if reproductive isolation (RI) barriers are not in place. We evaluated the importance of pre- and post-pollination RI barriers in three sympatric species of Achimenes (Gesneriaceae), including ecogeographic, phenological, floral isolation, self-pollination, and hybrid viability (fruit and seed set). We recorded geographic distribution throughout species ranges and assessed flowering phenology and pollinator visitation at one site in central Mexico. In the greenhouse, we measured floral traits involved in RI and quantified fruit and seed set for from self, intraspecific, and interspecific crosses. Ecogeographic barriers were important in RI, but under sympatry, phenological and floral barriers contributed more to total RI. Phenological RI varied between species and years, while floral RI was 100% effective at preventing interspecific visitation. Species showed differences in floral morphology, color, and scents associated with specialized pollination systems (A. antirrhina-hummingbirds, A. flava-bees, A. patens-butterflies); heterospecific visitation events were restricted to rare secondary pollinators. Hybrid crosses consistently yielded progeny in lower numbers than intraspecific crosses. This study indicated that neither autogamy nor early post-pollination barriers prevent interspecific pollen flow between Achimenes species. However, floral isolation, acting through a combination of attraction and reward traits, consistently ensures specificity of the pollination system. These results suggest that selection on floral traits to reduce the costs of hybrid progeny production may have played a role in evolution or maintenance of specialized pollination systems in Achimenes.

A comparison of reproductive isolation between two closely related oak species in zones of recent and ancient secondary contact

BackgroundMuch of the debate over the evolutionary consequences of hybridization on genetic divergence and speciation results from the breakdown or reinforcement of reproductive barriers in secondary hybrid zones. Among hybrid populations established for different lengths of time following secondary contact, stronger reproductive barriers are generally expected to occur in zones with longer contact. However, in plants no detailed investigation of recent and ancient zones of secondary contact has been conducted despite the importance of such a comparative study. Here, we compare pre- and postzygotic reproductive barriers between two closely related oak species, Quercus mongolica and Q. liaotungensis, in such a situation.ResultsThe recorded flowering times of both species overlapped in both contact zones. The fruit set at 10 and 30 days after interspecific hand pollination was not significantly lower than that after intraspecific pollination whenever Q. mongolica or Q. liaotungensis comprised the maternal parents in both populations. These results indicated that neither prezygotic phenological barriers nor interspecific incompatibility could have resulted in the reproductive isolation between the two species in both hybrid zones. However, the proportion of hybrid seeds produced by both species in the ancient zone was significantly lower than that recorded in the recent zone of secondary contact. In addition, the proportion of hybrid seeds simulated to form, assuming both random mating and an absence of postpollination barriers, was significantly higher than that detected in the ancient contact zone but not in the recent contact zone. These results suggest stronger early-acting postzygotic isolation between the two oak species in the ancient relative to the recent contact zone.ConclusionsOur comparative study demonstrated that postzygotic barriers during seed maturity were the main contributing factor to total reproductive isolation, particularly in the ancient contact zone, which aided species delimitation. In the recently formed secondary contact zone, pre- and postzygotic barriers were not well developed, and a high frequency of natural hybridization was evident. To our knowledge this study provides the first comparison of reproductive isolation between the ancient and recent secondary contact zones in plants and helps to clarify the evolutionary consequences of hybridization in a temporal context.

Reproductive isolation between Salvia elegans and S.fulgens, two hummingbird-pollinated sympatric sages.

The integrity of species in sympatric contact sites is dependent on the existence of reproductive isolating mechanisms, which restrict gene flow between them. However, we know little about the mechanisms that enable the coexistence of species with similar floral morphologies. Here, we evaluated several reproductive isolation barriers between Salvia elegans and S. fulgens, two sympatric sages with a similar ornithophilous floral syndrome, offering nectar as the main reward. Over 3 years, we evaluated broad-scale geographic isolation, floral phenologies and floral visitors as pre-pollination barriers, and fruit set, seed number and seed germination as post-pollination barriers. We found considerable geographic isolation and significant altitudinal differences between the two sages. The flowering period of both sages always overlapped extensively during the 3years of this study, but hummingbirds were highly specific, visiting one or the other Salvia species and showing aggressive territorial behaviour. Interspecific experimental crosses revealed that hybrid seeds might be formed although strong asymmetric barriers were found depending on the species acting as the maternal donor. Despite the low level of flowering asynchrony, reproductive isolation was remarkably high in the two sages. Geographic isolation and pollinator fidelity were the main factors responsible for maintaining species integrity. Despite an extensive review, we found very few studies quantifying the efficiency of isolation barriers in Neotropical plants or even the importance of hummingbirds as pollinators.

Impact of pre- and post-pollination barriers on pollen transfer and reproductive isolation among three sympatric Pedicularis (Orobanchaceae) species.

Pedicularis is the largest genus in the Orobanchaceae (>300) with many species co-occurring and co-blooming in subalpine to alpine meadows in the Himalayas. Although it is well known that different Pedicularis species place pollen on different parts of the same bumblebee's body, thus reducing interspecific pollen transfer, it is not known whether post-pollination components also contribute to reproductive isolation (RI). In this study, we quantified the individual strengths and absolute contributions of six pre- and post-pollination components of RI between three sympatric species in two pairs; Pedicularis gruina×Pedicularis tenuisecta (gru×ten) and Pedicularis comptoniifolia×Pedicularis tenuisecta (com×ten). All three Pedicularis species shared the same Bombus species. Individual foragers showed a high, but incomplete, floral constancy for each species. Therefore, pre-pollination barriers were potentially 'leaky' as Bombus species showed a low but consistent frequency of interspecific visitation. The RI strength of pre-pollination was lower in com×ten than in gru×ten. In contrast, post-pollination barriers completely blocked gene flow between both sets of species pairs. Two post-pollination recognition sites were identified. Late acting rejection of interspecific pollen tube growth occurred in com♀×ten♂, while seeds produced in bi-directional crosses of gru×ten failed to germinate. We propose that, although floral isolation based on pollen placement on pollinators in the genus Pedicularis is crucial to avoid interspecific pollen transfer, the importance of this mode of interspecific isolation may be exaggerated. Post-pollination barriers may play even larger roles for currently established populations of co-blooming and sympatric species in this huge genus in the Himalayas.

Self-incompatibility in Habranthus gracilifolius (Amaryllidaceae): pre- and post-pollination barriers

The evolution of self-incompatibility seems to be responsible for a great part of the evolutionary success of flowering plants by preventing self-fertilization and, hence, the effects of inbreeding. The recognition of incompatibility mechanisms allows us to explore their emergency and functionality and is particularly interesting in hermaphrodite plants due to the sexual conflict they experience. Here, we investigated some pre- and post-pollination barriers for self-reproduction in Habranthus gracilifolius Herb. In a natural population, we performed hand pollinations to check mating and incompatibility systems, measured the distance between anthers and stigma within flowers, and observed flower visitors. Our results evidence that this species is self-incompatible and reproduces via outcrossing (90% of crossed flowers set fruits). Self-pollen grains germinate on the stigma surface and are stopped at the beginning of the style, typical of gametophytic self-incompatibility. The species presents approach herkogamy–stigma is positioned in average 2.78 mm above stamens, which may reduce pollen self-interference. It is visited by a variety of insects, many acting as thieves. Our indirect measures of pollinator performance suggest that flower visitation frequency alone is not enough to evaluate visitors’ effects on fruit set. Herkogamy and gametophytic self-incompatibility prevent self-pollination and self-fertilization, respectively, and together with pollinators appropriated size and correct work of blossoms ensure the reproductive success of this species.

Does reproductive isolation reflect the segregation of color forms in Spiranthes sinensis (Pers.) Ames complex (Orchidaceae) in the Chinese Himalayas?

Isolation between species, or taxa sharing a common lineage, depends primarily on the relative strengths of various reproductive barriers. Previous studies on reproductive isolation between orchids emphasized mechanical and ethological barriers in flowers of species showing food and/or sexual mimicry. In this study, we investigated and quantified a series of prepollination and postpollination barriers between pink and white forms of Spiranthes sinensis sl, a nectar‐secreting complex. We generated ML trees based on trnS‐G and matK to explore phylogenetic relationships in this species complex. Spiranthes sinensis sl segregated from some other congeners, but the white form constituted a distinct clade in relation to the pink form. The white form secreted 2‐Phenylethanol as it is a single‐scent compound and was pollinated almost exclusively by native, large‐bodied Apis cerana and Bombus species (Apidae). Apis cerana showed a high floral constancy to this form. The scentless, pink form was pollinated primarily by smaller bees in the genera Ceratina (Apidae), and members of the family Halictidae, with infrequent visits by A. cerana and Bombus species. Fruit set and the production of large embryos following interform pollination treatments were significantly lower compared to intraform pollination results for the white form. Our results suggested that pollinator isolation, based on color and scent cues, may result in greater floral constancy in white populations when both forms are sympatric as two different, guilds of pollinators forage selectively preventing or reducing prospective gene flow. Postpollination barriers appear weaker than prepollination barriers but they also play a role in interform isolation, especially in the white form. Our findings suggest that floral color forms in S. sinensis do not represent an unbalanced polymorphism. Interpretations of the evolutionary status of these forms are discussed.

Interspecific Divergence of Two Sinalliaria (Brassicaceae) Species in Eastern China.

How endemic species originated in eastern Asia has interested botanists for a long time. In this study, we combined experimental and computational modeling approaches to examine the morphological and genetic divergence and reproductive isolation of two tentative species of Sinalliaria (Brassicaceae) endemic to eastern China, S. limprichtiana and S. grandifolia. Most of the examined morphological characters (including hairs of leaf blades and stems, corolla length and width, and flower stalk length) were well-delineated between two species at the same ploidy level, and there was clear evidence of reproductive isolation between them (mainly due to post-pollination barriers) in the common garden environment. There were also strong and consistent divergences in the population genetic data. Coalescent simulations based on sequence variation of the nuclear genes suggest that interspecific divergence began during the Pleistocene when the climate oscillated in eastern Asia. Gene flow between two species appears to have been very limited and asymmetrical. Our results suggested that both species are well-differentiated and that the fast divergence between them might have been together shaped by both stochastic processes and habitat selection pressures.