ABSTRACT• Heterostyly is a sex polymorphism that has challenged evolutionary biologists ever since Darwin. One of the lineages where heterostyly and related stylar conditions appear more frequently is Linum (Linaceae). This group is particularly suitable for testing competing hypotheses about ancestral and transitional stages on the evolutionary building up of heterostyly.• We generated a phylogeny of Linum based on extensive sampling and plastid and nuclear DNA sequences, and used it to trace the evolution of character states of style polymorphism. We also revised available data on pollination, breeding systems, and polyploidy to analyse their associations.• Our results supported former phylogenetic hypotheses: the paraphyly of Linum and the non-monophyly of current taxonomic sections. Heterostyly was common in the genus, but appeared concentrated in the Mediterranean Basin and the South African Cape. Ancestral character state reconstruction failed to determine a unique state as the most probable condition for style polymorphism in the genus. In contrast, approach herkogamy was resolved as ancestral state in some clades, giving support to recent hypotheses. Some traits putatively related to heterostyly, such as life history and polyploidy, did show marginal or non-significant phylogenetic correlation, respectively. Although pollinator data are limited, we suggest that beeflies are associated with specific cases of heterostyly.• The consistent association between style polymorphism and heteromorphic incompatibility points to ecological factors as drivers of the multiple evolution of style polymorphism in Linum. Albeit based on limited evidence, we hypothesised that specialised pollinators and lack of mating opportunities drive evolution of style polymorphism and loss of the polymorphism, respectively.

Summary1. In hermaphroditic flowers, reciprocal herkogamy, for example heterostyly, enhances pollen transfer between floral morphs (disassortative pollination) while avoiding self-interference between sexual organs. By contrast, disassortative pollination might be compromised in style-dimorphic flowers, which lack perfect reciprocity between the two floral morphs. This suboptimal functioning has been considered to explain why stylar dimorphism is rare in nature.2. Some style-polymorphic species receive a wide array of floral visitors, including long-tongued insects that feed on nectar and short-tongued insects that feed on pollen. Differences in the morphology and behaviour of these two insect types could be manifested as different pollination patterns in each floral morph.3. By observing individual pollinators and pollen deposition and delivery in small field-based experimental arrays with emasculated and intact flowers, we studied pollination patterns mediated by different insect types (long-and short-tongued) in the two floral morphs (long-and short-styled) of the style-dimorphic Narcissus papyraceus. We investigated the patterns of pollen transfer between and within style morphs in cross-pollinations, as well as self-pollination and pollen removal rates, for each floral morph mediated by each insect type.4. Long-tongued insects were efficient pollinators of both floral morphs as they removed little pollen from the anthers but deposited comparatively large amounts on the stigmas. Although disassortative pollen transfer and assortative pollen transfer were equally high to the long-styled morph, the former prevailed in the short-styled morph. Short-tongued insects removed large amounts of pollen from the anthers but deposited only a few pollen grains on the long-styled stigmas and a negligible number of grains on short-styled stigmas regardless of the morph of the donor. 5.Synthesis. In this study, we provide empirical support for the hypothesis that, under the action of long-tongued pollinators, pollen transfer patterns in the stylar-dimorphic Narcissus papyraceus resemble in part those of heterostylous species. In addition, we found that short-tongued insects act mostly as pollen thieves, thereby limiting the male fitness of both style morphs, besides depleting the female fitness of S-morph plants. In view of these results, we propose that the differing pollination efficiencies of floral visitors, in addition to their frequency, are key in determining the morph ratio of populations in this Narcissus.

Aim  Our aims were: (1) to reconstruct the phylogenetic relationships of daffodils (Narcissus), focusing on the lowland subgenus Hermione and the mountain section Apodanthi; (2) to estimate the temporal setting of diversification; (3) to reconstruct the migration patterns of the lineages; and (4) to examine the microevolutionary differentiation of the wide‐ranging Narcissus tazetta group across the Mediterranean. Location  The Mediterranean Basin. Methods  Plastid (trnT–L, trnL–F and ndhF) sequences were obtained from 63 populations representing 23 species of Narcissus and combined with published data from 16 species. Phylogenetic relationships and dating were inferred by Bayesian analysis based on geological events and divergence estimates of closely related taxa. A dispersal–extinction–cladogenesis analysis was performed using maximum likelihood methods to infer ancestral geographical distributions, and phylogeographical reconstruction was performed using coalescence analysis. Results  Subgenus Hermione is not recognized as a monophyletic group because two of the nine species were found to have a close relationship with the subgenus Narcissus. The results on section Apodanthi confirmed previous findings of its monophyly and phylogenetic relationships within this mountain group. Molecular dating and ancestral range reconstructions suggest that the ancestor of Narcissus originated in the Iberian Peninsula during the Late Oligocene–Early Miocene. Eastward expansion of the lineage range proceeded from the western Mediterranean and involved colonization of mountain ranges in northern Africa. The phylogeography of the N. tazetta group revealed a widespread distribution of certain haplotypes, suggesting wide dispersal and a high level of colonization in the Mediterranean Basin. Main conclusions  Our study points to the role of three key historical events in Narcissus diversification: tectonic shifts of the Alboran domain in the western Mediterranean, the Messinian salinity crisis, and the onset of the Mediterranean climate followed by periods of repeated glaciation. Diversification of section Apodanthi probably resulted from allopatric speciation, while subgenus Hermione may have shown more sympatric speciation and high dispersal, despite the lack of apparent adaptations to long‐distance dispersal. This is best exemplified by the presence of both ancestral and recent haplotypes of N. tazetta across the Mediterranean.

The style morph ratio changed gradually, whereas perianth trait variation showed abrupt changes with two morphotypes across the range. The positive relationship between the visitation rate of short-tongued pollinators and the decrease of the short-style morph supports our initial hypothesis. The results highlight the importance of different pollinators in determining the presence of style polymorphism.

Darwin's early work on heterostyly and related style polymorphisms (the presence of two or three style morphs within a population) generated much interest to understand how precise interactions between ecological and genetic mechanisms influence the evolution of floral diversity. Here we tested three key hypotheses proposed to explain the evolution of heterostyly: (i) the presence of self-incompatibility; (ii) the role of pollinators in promoting dissasortative mating; and (iii) floral architecture, which restricts pollinators' movements and ensures more exact pollen deposition on their bodies. We combined data from experiments, field observations, and published studies to test whether evolution of style polymorphism in Narcissus is driven by the incompatibility system, pollinator guilds, or floral architecture, within a phylogenetic framework. Neither differences in pollinator environment nor the presence of genetic self-incompatibility were correlated with presence of style polymorphism. However, our results indicate that the evolution of style polymorphism was driven by the presence of a narrow and long floral tube. K E Y W O R D S :Evolutionary transitions, floral architecture, heterostyly, incompatibility system, pollinators.Heterostyly and related style polymorphisms have fascinated biologists since Darwin provided an evolutionary explanation of the mechanisms suggesting that they serve to promote outcrossing in hermaphroditic plants (Darwin 1877). Heterostyly is characterized by the presence of two (distyly) or three (tristyly) flower morphs with reciprocal displacement in the height of the stigmas and the anthers within a flower. Style dimorphism is a polymorphism related with heterostyly, where two stylar morphs exist without concomitant reciprocal variation in stamens. Evolutionary models suggest that stylar dimorphism is an intermediate step in the transition from style monomorphism to distyly (Charlesworth and Charlesworth 1979;Lloyd and Webb 1992;Graham and Barrett 2004); however, such an intermediate step is present in comparatively few taxonomical groups. Heterostyly is a convergent trait present in at least 28 angiosperm families (Barrett and Shore 2008), and the long-standing interest in this style polymorphism has generated a diversity of hypotheses involving different scenarios and selective forces to explain its evolution