Sex-dependent phenological responses to climate vary across species' ranges.

Anthropogenic climate change has significantly altered the flowering times (i.e., phenology) of plants worldwide, affecting their reproduction, survival, and interactions. Recent studies utilizing herbarium specimens have uncovered significant intra- and inter-specific variation in flowering phenology and its response to changes in climate but have mostly been limited to animal-pollinated species. Thus, despite their economic and ecological importance, variation in phenological responses to climate remain largely unexplored among and within wind-pollinated dioecious species and across their sexes. Using both herbarium specimens and volunteer observations of cottonwood (Populus) species, we examined how phenological sensitivity to climate varies across species, their ranges, sexes, and phenophases. The timing of flowering varied significantly across and within species, as did their sensitivity to spring temperature. In particular, male flowering generally happened earlier in the season and was more sensitive to warming than female flowering. Further, the onset of flowering was more sensitive to changes in temperature than leaf out. Increased temporal gaps between male and female flowering time and between the first open flower date and leaf out date were predicted for the future under two climate change scenarios. These shifts will impact the efficacy of sexual reproduction and gene flow among species. Our study demonstrates significant inter- and intra-specific variation in phenology and its responses to environmental cues, across species' ranges, phenophases, and sex, in wind-pollinated species. These variations need to be considered to predict accurately the effects of climate change and assess their ecological and evolutionary consequences.

Pollination efficiency and the pollen-ovule ratio.

Pollination presents a risky journey for pollen grains. Pollen loss is sometimes thought to favour greater pollen investment to compensate for the inefficiency of transport. Sex allocation theory, to the contrary, has consistently concluded that postdispersal loss should have no selective effect on investment in either sex function. But the intuitively appealing compensation idea continues to be raised despite the lack of theoretical endorsement. We address the theoretical issue with a model that directly represents pollen loss (and ovule loss through floral demise or loss of receptivity) as rate-dependent dynamical processes. These loss rates can be varied to examine the effect of pollination efficiency on optimal sex allocation. Pollen-ovule ratios follow from the sex allocation based on the resource costs of pollen and ovule production. This model confirms conventional findings that pollen loss should have essentially no effect on sexual resource allocation in large, panmictic populations. Pollen limitation of seed set does not alter this conclusion. These results force us to rethink the empirical association of pollination efficiency with low pollen-ovule ratios. This pattern could arise if efficient pollen transport commonly results in stigmatic deposition of cohorts of related pollen. Empirical evidence of correlated paternity supports this explanation.

Global wind patterns shape genetic differentiation, asymmetric gene flow, and genetic diversity in trees.

Wind disperses the pollen and seeds of many plants, but little is known about whether and how it shapes large-scale landscape genetic patterns. We address this question by a synthesis and reanalysis of genetic data from more than 1,900 populations of 97 tree and shrub species around the world, using a newly developed framework for modeling long-term landscape connectivity by wind currents. We show that wind shapes three independent aspects of landscape genetics in plants with wind pollination or seed dispersal: populations linked by stronger winds are more genetically similar, populations linked by directionally imbalanced winds exhibit asymmetric gene flow ratios, and downwind populations have higher genetic diversity. For each of these distinct hypotheses, partial correlations between the respective wind and genetic metrics (controlling for distance and climate) are positive for a significant majority of wind-dispersed or wind-pollinated genetic data sets and increase significantly across functional groups expected to be increasingly influenced by wind. Together, these results indicate that the geography of both wind strength and wind direction play important roles in shaping large-scale genetic patterns across the world's forests. These findings have implications for various aspects of basic plant ecology and evolution, as well as the response of biodiversity to future global change.

5S-IGS rDNA in wind-pollinated trees (Fagus L.) encapsulates 55 million years of reticulate evolution and hybrid origins of modern species.

Standard models of plant speciation assume strictly dichotomous genealogies in which a species, the ancestor, is replaced by two offspring species. The reality in wind-pollinated trees with long evolutionary histories is more complex: species evolve from other species through isolation when genetic drift exceeds gene flow; lineage mixing can give rise to new species (hybrid taxa such as nothospecies and allopolyploids). The multi-copy, potentially multi-locus 5S rDNA is one of few gene regions conserving signal from dichotomous and reticulate evolutionary processes down to the level of intra-genomic recombination. Therefore, it can provide unique insights into the dynamic speciation processes of lineages that diversified tens of millions of years ago. Here, we provide the first high-throughput sequencing (HTS) of the 5S intergenic spacers (5S-IGS) for a lineage of wind-pollinated subtropical to temperate trees, the Fagus crenata - F. sylvatica s.l. lineage, and its distant relative F. japonica. The observed 4963 unique 5S-IGS variants reflect a complex history of hybrid origins, lineage sorting, mixing via secondary gene flow, and intra-genomic competition between two or more paralogous-homoeologous 5S rDNA lineages. We show that modern species are genetic mosaics and represent a striking case of ongoing reticulate evolution during the past 55 million years.

Pollen production in olive cultivars and its interannual variability.

BACKGROUND AND AIMS: Olive (Olea europaea subsp. europaea var. europaea) is the most extensively cultivated fruit crop worldwide. It is considered a wind-pollinated and strictly outcrossing crop. Thus, elevated pollen production is crucial to guarantee optimum fruit set and yield. Despite these facts, the variability of pollen production within the cultivated olive has been scarcely studied. This study aimed to characterize this feature by analysing a representative set of worldwide olive cultivars. METHODS: We evaluated the average number of pollen grains per anther in 57 principal cultivars over three consecutive years. We applied a standard generalized linear model (GLM) approach to study the influence of cultivar, year and the previous year's fruit load on the amount of pollen per anther. Additionally, the K-means method was used for cluster analysis to group cultivars based on their pollen production capacity. KEY RESULTS: Pollen production per anther was highly variable among olive cultivars. The cultivar significantly accounted for 51.3 % of the variance in pollen production and the year for 0.3 %. The interaction between the two factors explained 8.4 % of the variance, indicating that not all cultivars were equally stable in producing pollen across the years. The previous year's fruit load and its interaction with the year were significant, but barely accounted for 1.5 % of the variance. Olive cultivars were classified into four clusters according to their capacity to produce pollen. Interestingly, the fourth cluster was composed of male-sterile cultivars, which presumably share this character by inheritance. CONCLUSIONS: Pollen production per anther varied extensively within the cultivated olive. This variation was mainly driven by the cultivar and its interaction with the year. The differential capacity of olive cultivars to produce pollen should be considered not only for designing new orchards but also gardens where this species is used as an ornamental.

Flower morphology as a predictor of pollination mode in a biotic to abiotic pollination continuum.

BACKGROUND AND AIMS: Wind pollination has evolved repeatedly in flowering plants, yet the identification of a wind pollination syndrome as a set of integrated floral traits can be elusive. Thalictrum (Ranunculaceae) comprises temperate perennial herbs that have transitioned repeatedly from insect to wind pollination while also exhibiting mixed pollination, providing an ideal system to test for evolutionary correlation between floral morphology and pollination mode in a biotic to abiotic continuum. Moreover, the lack of floral organ fusion across this genus allows testing for specialization to pollination vectors in the absence of this feature. METHODS: We expanded phylogenetic sampling in the genus from a previous study using six chloroplast loci, which allowed us to test whether species cluster into distinct pollination syndromes based on floral morphology. We then used multivariate analyses on floral traits followed by ancestral state reconstruction of the emerging flower morphotypes and determined whether these traits are evolutionarily correlated under a Bayesian framework with Brownian motion. KEY RESULTS: Floral traits fell into five distinct clusters, which were reduced to three after considering phylogenetic relatedness and were largely consistent with flower morphotypes and associated pollination vectors. Multivariate evolutionary analyses found a positive correlation between the lengths of floral reproductive structures (styles, stigmas, filaments and anthers). Shorter reproductive structures tracked insect-pollinated species and clades in the phylogeny, whereas longer structures tracked wind-pollinated ones, consistent with selective pressures exerted by biotic vs. abiotic pollination vectors, respectively. CONCLUSIONS: Although detectable suites of integrated floral traits across Thalictrum were correlated with wind or insect pollination at the extremes of the morphospace distribution, a presumed intermediate, mixed pollination mode morphospace was also detected. Thus, our data broadly support the existence of detectable flower morphotypes from convergent evolution underlying the evolution of pollination mode in Thalictrum, presumably via different paths from an ancestral mixed pollination state.

Estimation of pollen dispersal distance in Job's tears (Coix lacryma-jobi L.) by using red leaf sheath as a morphological marker.

Job's tears (Coix lacryma-jobi L.) is grown widely in Asian countries and a crop that can fertilize with own pollen and pistils. The grains are used not only for food but also for medicinal purposes. The grain of many cultivars contains glutinous endosperm; only grains with this glutinous endosperm are suitable for use as medicine in Japan. Many wild types have non-glutinous endosperm and can easily cross with cultivar under natural environmental conditions. Because the non-glutinous endosperm trait is dominant to that of glutinous endosperm, F1 seeds produced by crosses between a cultivar and a wild type have non-glutinous endosperm. To reduce the rate of unwanted crosses, we investigated the pollen dispersal distance by using a red leaf sheath as a morphological marker. When plants were cultivated in rows 70 cm apart, the crossing rate was about 25%-35%. As the distance increased, the crossing rate decreased at a rate that could be fitted to a power approximation in fields without intervening plants and to an exponential equation in fields with intervening plants. Our data could be used as guidelines for preventing unwanted crossing with wild types when growing cultivars.

Simulated herbivory enhances leaky sex expression in the dioecious herb Mercurialis annua.

BACKGROUND AND AIMS: Plant reproductive traits are widely understood to be responsive to the selective pressures exerted by pollinators, but there is also increasing evidence for an important role for antagonists such as herbivores in shaping these traits. Many dioecious species show leaky sex expression, with males and females occasionally producing flowers of the opposite sex. Here, we asked to what extent leakiness in sex expression in Mercurialis annua (Euphorbiaceae) might also be plastically responsive to simulated herbivory. This is important because enhanced leakiness in dioecious populations could lead to a shift in both the mating system and in the conditions for transitions between combined and separate sexes. METHODS: We examined the effect of simulated herbivory on the sexual expression of males and females of M. annua in two experiments in which different levels of simulated herbivory led to enhanced leakiness in both sexes. KEY RESULTS: We showed that leaky sex expression in both males and females of the wind-pollinated dioecious herb M. annua is enhanced in response to simulated herbivory, increasing the probability for and the degree of leakiness in both sexes. We also found that leakiness was greater in larger females but not in larger males. CONCLUSIONS: We discuss hypotheses for a possible functional link between herbivory and leaky sex expression, and consider what simulated herbivory-induced leakiness might imply for the evolutionary ecology of plant reproductive systems, especially the breakdown of dioecy and the evolution of hermaphroditism.

Pollen-insect interaction meta-networks identify key relationships for conservation in mosaic agricultural landscapes.

Flower visitors use different parts of the landscape through the plants they visit, however these connections vary within and among land uses. Identifying which flower-visiting insects are carrying pollen, and from where in the landscape, can elucidate key pollen-insect interactions and identify the most important sites for maintaining community-level interactions across land uses. We developed a bipartite meta-network, linking pollen-insect interactions with the sites they occur in. We used this to identify which land-use types at the site- and landscape-scale (within 500 m of a site) are most important for conserving pollen-insect interactions. We compared pollen-insect interactions across four different land uses (remnant native forest, avocado orchard, dairy farm, rotational potato crop) within a mosaic agricultural landscape. We sampled insects using flight intercept traps, identified pollen carried on their bodies and quantified distinct pollen-insect interactions that were highly specialized to both natural and modified land uses. We found that sites in crops and dairy farms had higher richness of pollen-insect interactions and higher interaction strength than small forest patches and orchards. Further, many interactions involved pollinator groups such as flies, wasps, and beetles that are often under-represented in pollen-insect network studies, but were often connector species in our networks. These insect groups require greater attention to enable wholistic pollinator community conservation. Pollen samples were dominated by grass (Poaceae) pollen, indicating anemophilous plant species may provide important food resources for pollinators, particularly in modified land uses. Field-scale land use (within 100 m of a site) better predicted pollen-insect interaction richness, uniqueness, and strength than landscape-scale. Thus, management focused at smaller scales may provide more tractable outcomes for conserving or restoring pollen-insect interactions in modified landscapes. For instance, actions aimed at linking high-richness sites with those containing unique (i.e., rare) interactions by enhancing floral corridors along field boundaries and between different land uses may best aid interaction diversity and connectance. The ability to map interactions across sites using a meta-network approach is practical and can inform land-use planning, whereby conservation efforts can be targeted toward areas that host key interactions between plant and pollinator species.

Flowering synchrony drives reproductive success in a wind-pollinated tree.

Synchronised and quasi-periodic production of seeds by plant populations, known as masting, is implicated in many ecological processes, but how it arises remains poorly understood. Flowering and pollination dynamics are hypothesised to provide the mechanistic link for the observed relationship between weather and population-level seed production. We report the first experimental test of the phenological synchrony hypotheses as a driver of pollen limitation in mast seeding oaks (Quercus ilex). Higher flowering synchrony yielded greater pollination efficiency, which resulted in 2-fold greater seed set in highly synchronised oaks compared to asynchronous individuals. Pollen addition removed the negative effect of asynchronous flowering on seed set. Because phenological synchrony operates through environmental variation, this result suggests that oak masting is synchronised by exogenous rather than endogenous factors. It also points to a mechanism by which changes in flowering phenology can affect plant reproduction of mast-seeding plants, with subsequent implications for community dynamics.

Influence of local density and sex ratio on pollination in an ambophilous flowering plant.

PREMISE: Variation in local density and sex ratio in dioecious plants can affect mating success through the actions of pollen vectors, principally generalist insects or wind. Increased density and male-biased sex ratios should promote pollen transfer and seed production, but their combined effects have not been investigated for ambophilous species, which exhibit both insect and wind pollination. METHODS: We manipulated density (low vs. high) and sex ratio (1:1 vs. 3:1 male-biased) in arrays of dioecious ambophilous Thalictrum pubescens. We quantified visitation rates and foraging times to examine whether pollinators exhibited sex-specific preferences and determined the seed set of arrays. RESULTS: Pollinators visited more plants per foraging bout at high than low density. Visitation rates and foraging times of visitors were greater for male than for female plants but did not depend on the density or sex ratio of arrays. However, whereas solitary bees displayed a strong preference for males, hover flies were indifferent to plant sex phenotype. Solitary bees also visited significantly more plants per foraging bout than hover flies. There was a significant interaction between density and sex ratio on seed set. At low density, seed set was greater for 3:1 than for 1:1 arrays, but at high density the opposite pattern occurred. CONCLUSIONS: The demographic factors we investigated had complex influences on pollinator foraging behavior and patterns of seed set. Several factors may explain our results, including the influence of density and sex ratio on pollen export from arrays, grooming by pollinators, and the contribution of wind pollination.

Variation in sexual dimorphism in a wind-pollinated plant: the influence of geographical context and life-cycle dynamics.

Understanding the mechanisms causing phenotypic differences between females and males has long fascinated evolutionary biologists. An extensive literature exists on animal sexual dimorphism but less information is known about sex differences in plants, particularly the extent of geographical variation in sexual dimorphism and its life-cycle dynamics. Here, we investigated patterns of genetically based sexual dimorphism in vegetative and reproductive traits of a wind-pollinated dioecious plant, Rumex hastatulus, across three life-cycle stages using open-pollinated families from 30 populations spanning the geographic range and chromosomal variation (XY and XY1 Y2 ) of the species. The direction and degree of sexual dimorphism was highly variable among populations and life-cycle stages. Sex-specific differences in reproductive function explained a significant amount of temporal change in sexual dimorphism. For several traits, geographical variation in sexual dimorphism was associated with bioclimatic parameters, likely due to the differential responses of the sexes to climate. We found no systematic differences in sexual dimorphism between chromosome races. Sex-specific trait differences in dioecious plants largely result from a balance between sexual and natural selection on resource allocation. Our results indicate that abiotic factors associated with geographical context also play a role in modifying sexual dimorphism during the plant life-cycle.

Comparative analysis of pollen release biomechanics in Thalictrum: implications for evolutionary transitions between animal and wind pollination.

Transitions from animal to wind pollination have occurred repeatedly in flowering plants, driven by structural and biomechanical modifications to flowers. But the initial changes promoting wind pollination are poorly understood, especially those required to release pollen into airflows - the critical first stage of wind pollination. Using a wind tunnel, we performed a comparative study of pollen release biomechanics in 36 species of animal- and wind-pollinated Thalictrum. We quantified pollination syndromes and stamen natural frequency (fn ), a key vibration parameter, to determine if floral traits reliably predicted pollen release probability. We then investigated if pollen release was caused by wind-induced resonance vibration of stamens. We detected wind-induced stamen resonance in 91% of species and a strong effect of stamen acceleration on pollen release, inversely driven by fn . However, unlike fn , pollination syndromes did not reliably predict the probability of pollen release among species. Our results directly link fn to the capacity of stamens to release pollen by wind and suggest that structural mechanisms reducing fn are likely to be important for initiating transitions from animal to wind pollination. Our inability to predict the probability of pollen release based on pollination syndromes suggests diverse phenotypic trajectories from animal to wind pollination.

Anatomy solves the puzzle of explosive pollen release in wind-pollinated urticalean rosids.

PREMISE OF THE STUDY: This study details the unusual synorganization of the staminate flower in wind-pollinated urticalean rosids to add the missing pieces that complete the puzzle of the explosive mechanism of pollen release in this group. METHODS: Flower buds and flowers were analyzed using light and scanning electron microscopy. KEY RESULTS: The pistillode, stamens, and sepals form a floral apparatus that explosively releases pollen to be carried by the wind. The anthers dehisce when the stamens are still inflexed on the floral bud and are enveloped by the sepals and supported by an inflated pistillode. The distension of the filaments presses the pistillode, which decreases the pressure exerted on the anthers by releasing the air accumulated internally through its apical orifice. The extended filaments and the dehiscent free anthers move rapidly outward from the center of the flower. This movement of the filaments is then blocked by the robust basally united sepals, which causes a rapid inversion of the anther position, thus hurling the pollen grains far from the flower. The pollen grains are released grouped by the mucilage produced in high quantity in the cells found in all floral organs. CONCLUSIONS: The anatomical structure of the pistillode and the finding of mucilaginous cells are the main features that help in the understanding the explosive mechanism of pollen release in urticalean rosids. The pistillode can be considered an exaptation because it was evolved later to provide a new role in the plant, optimizing male fitness.

Does sexual dimorphism exist in flowering phenology traits in anemophilous dioecious species? A test with Rumex acetosa.

PREMISE: Sexual dimorphism in flowering phenology traits may have evolved under sexual selection and vector-mediated selection. The conspicuous sexual dimorphism and sex-specific selection pressures in flowering phenology traits have been investigated mainly in entomophilous dioecious plants, whereas little is known about this in anemophilous plants. METHODS: We examined sexual dimorphism in flowering onset, flowering peak, flowering duration, maximum proportion of open flowers per inflorescence branch, maximum proportion of newly opening flowers on a given date per branch, and longevity of individual flowers in natural Rumex acetosa populations. Correlations between flowering phenology traits and the degree of flowering overlap with the opposite sex were examined. We also tested whether the overlap of female flowering with male flowering enhanced seed set in female plants. RESULTS: Little sexual dimorphism was observed in flowering onset, peak, duration, and maximum proportion of newly opening flowers. Females had greater floral longevity and greater maximum proportion of open flowers than males. Flowering overlap with the opposite sex significantly increased with the maximum proportion of newly opening flowers and decreased with temporal deviation in the flowering peak in both sexes. Females with greater flowering overlap with males set more seeds in two of the three study populations. CONCLUSIONS: In wind-pollinated R. acetosa, little sexual dimorphism in phenological traits may have evolved to achieve synchronous flowering with the opposite sex. Our results suggest that, in angiosperms, not only common selection but also anemophily-specific selection may shape little sexual dimorphism in R. acetosa, unlike in entomophilous plants.

Divergent selection on the biomechanical properties of stamens under wind and insect pollination.

Wind pollination has evolved from insect pollination in numerous angiosperm lineages and is associated with a characteristic syndrome of morphological traits. The traits initiating transitions to wind pollination and the ecological drivers involved are poorly understood. Here, we examine this problem in Thalictrum pubescens, an ambophilous (insect and wind pollination) species that probably represents a transitional state in the evolution of wind pollination. We investigated wind-induced pollen release by forced harmonic motion by measuring stamen natural frequency ( fn), a key vibration parameter, and its variability among nine populations. We assessed the repeatability of fn over consecutive growing seasons, the effect of this parameter on pollen release in a wind tunnel, and male reproductive success in the field using experimental manipulation of the presence or absence of pollinators. We found significant differences among populations and high repeatability within genotypes in fn. The wind tunnel assay revealed a strong negative correlation between fn and pollen release. Siring success was greatest for plants with lower fn when pollinators were absent, but this advantage diminished when pollinators were present. Our biomechanical analysis of the wind-flower interface has identified fn as a key trait for understanding early stages in the transition from insect to wind pollination.

Effects of heterospecific pollen from a wind-pollinated and pesticide-treated plant on reproductive success of an insect-pollinated species.

PREMISE OF THE STUDY: Studies on the effects of heterospecific pollen (HP) transfer have been focused mainly on insect-pollinated species, despite evidence of insect visitation to wind-pollinated species and transfer of their pollen onto stigmas of insect-pollinated plants. Thus, the potential consequences of HP transfer from wind-pollinated species remain largely unknown. Furthermore, accumulation of pesticide residues in pollen of wind-pollinated crops has been documented, but its potential effects on wild plant species via HP transfer have not been tested. METHODS: We evaluated the effect of wind-dispersed Zea mays pollen on pollen tube growth of the insect-pollinated Mimulus nudatus via hand pollinations. We further evaluated whether pesticide-contaminated Z. mays pollen has larger effects on M. nudatus pollen success than non-contaminated Z. mays pollen. KEY RESULTS: We found a significant negative effect of Z. mays pollen on M. nudatus pollen tube growth even when deposited in small amounts. However, we did not observe any difference in the magnitude of this effect between pesticide-laden Z. mays pollen and non-contaminated Z. mays pollen. CONCLUSIONS: Our results suggest that wind-pollinated species can have negative effects as HP donors on insect-pollinated recipients. Thus, their role in shaping co-flowering interactions for wind- and insect-pollinated species deserves more attention. Although we did not find evidence that pesticide contamination increased HP effects, we cannot fully rule out the existence of such an effect, because pollen load and thus the pesticide dose applied to stigmas was low. This result should be confirmed using other HP donors and across a range of HP loads, pesticide types, and concentrations.

Evolutionary drivers of mast-seeding in a long-lived desert shrub.

PREMISE OF THE STUDY: The evolutionary drivers and proximal regulators of mast-seeding are well understood for species of mesic environments, but how these regulators interact with high spatial and interannual variability in growing-season precipitation for a masting species in a desert environment has never been examined. METHOD: We followed flowering and seed production in 16 populations of the North American desert shrub blackbrush (Coleogyne ramosissima) from contrasting environments across its range over an 11-year period to determine patterns of interannual reproductive output variation. KEY RESULT: Patterns of reproductive output in blackbrush did not track current growing season precipitation, but instead were regulated by prior-year weather cues. The strength of the response to the masting cue depended on habitat quality, with higher mean reproductive output, shorter intervals between years of high seed production, and lower CVp at more favorable sites. Wind pollination efficiency was demonstrated to be an important evolutionary driver of masting in blackbrush, and satiation of heteromyid seed predator-dispersers was supported as an evolutionary driver based on earlier studies. CONCLUSIONS: Both the evolutionary drivers and proximal regulators of masting in blackbrush are similar to those demonstrated for masting species of mesic environments. Relatively low synchrony across populations in response to regional masting cues occurs at least partly because prior-year environmental cues can trigger masting efforts in years with resource limitation due to suboptimal precipitation, especially in more xeric low-elevation habitats.

Timing and consequences of recurrent polyploidy in meadow-rues (thalictrum, ranunculaceae).

The discovery of ancient whole-genome duplications in eukaryotic lineages has renewed the interest in polyploidy and its effects on the diversification of organisms. Polyploidy has large-scale effects on both genotype and phenotype and has been linked to the evolution of genome size, dioecy, and changes in ecological interactions, such as pollinator visitation. Here, we take a molecular systematics approach to examine the evolution of polyploidy in the plant genus Thalictrum (Ranunculaceae) and test its correlation to changes in genome size, sexual system, and pollination mode. Thalictrum is an ideal study system due to its extensive ploidy range and floral diversity. Phylogenetic analyses were used for character reconstructions, correlation tests, and dating estimates. Our results suggest that polyploidization occurred frequently and recently in the evolution of Thalictrum, mostly within the last 10.6-5.8 My, coinciding with the diversification of particular clades. In spite of an overall trend of genomic downsizing accompanying polyploidy in angiosperms and proportional increases observed at finer scales, our genome size estimates for Thalictrum show no correlation with chromosome number. Instead, we observe genomic expansion in diploids and genomic contraction in polyploids with increased age. Additionally, polyploidy is not correlated with dioecy in Thalictrum; therefore, other factors must have influenced the evolution of separate sexes in this group. A novel finding from our study is the association of polyploidy with shifts to wind pollination, in particular, during a time period of global cooling and mountain uplift in the Americas.

Segregation of male-sterility alleles across a species boundary.

Hybrid zones may serve as bridges permitting gene flow between species, including alleles influencing the evolution of breeding systems. Using greenhouse crosses, we assessed the likelihood that a hybrid zone could serve as a conduit for transfer of nuclear male-sterility alleles between a gynodioecious species and a hermaphroditic species with very rare females in some populations. Segregation patterns in progeny of crosses between rare females of hermaphroditic Schiedea menziesii and hermaphroditic plants of gynodioecious Schiedea salicaria heterozygous at the male-sterility locus, and between female S. salicaria and hermaphroditic plants from the hybrid zone, were used to determine whether male-sterility was controlled at the same locus in the parental species and the hybrid zone. Segregations of females and hermaphrodites in approximately equal ratios from many of the crosses indicate that the same nuclear male-sterility allele occurs in the parent species and the hybrid zone. These rare male-sterility alleles in S. menziesii may result from gene flow from S. salicaria through the hybrid zone, presumably facilitated by wind pollination in S. salicaria. Alternatively, rare male-sterility alleles might result from a reversal from gynodioecy to hermaphroditism in S. menziesii, or possibly de novo evolution of male sterility. Phylogenetic analysis indicates that some species of Schiedea have probably evolved separate sexes independently, but not in the lineage containing S. salicaria and S. menziesii. High levels of selfing and expression of strong inbreeding depression in S. menziesii, which together should favour females in populations, argue against a reversal from gynodioecy to hermaphroditism in S. menziesii.