Search

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.

Sexual reproduction in heterostylous populations may be vulnerable to demographic conditions because of the small number of mating types in populations. Here, we investigate mating and fertility under natural and experimental conditions in tristylous Lythrum salicaria L., an invasive species that exhibits a wide range of floral morph ratios and demographic contexts. We grew 147 open-pollinated seed families from six populations with different morph structures to estimate intermorph mating (d). In a field experiment, we used progeny ratios from 47 spatially isolated individuals to estimate d, and measured the intensity of pollen limitation experienced by the morphs. The M- and S-morphs experienced high rates of d, regardless of population size or morph ratio. Estimates for the L-morph revealed low levels of intramorph mating in three dimorphic and two trimorphic populations, but near complete intramorph mating in a monomorphic population. Despite high levels of intermorph mating in the field experiment, the morphs experienced significant pollen limitation of fruit and seed set, but this did not differ in intensity among the morphs. Our field experiment demonstrates that although plant isolation was associated with pollen limitation of seed set, “long-distance” bee-mediated pollen flow served to maintain intermorph mating. Tristyly in L. salicaria is remarkably robust to the demographic variation associated with colonization.

Premise of research. Angiosperms are commonly classified as self-incompatible or self-compatible. This dichotomy has influenced research on the ecological and demographic consequences of colonizati...

Premise of the study Darwin proposed that the reciprocal arrangement of anthers and stigmas in heterostylous plants promotes cross-pollination through pollen segregation on pollinators' bodies. The floral tube in most heterostylous species constrains the feeding posture of pollinators determining the site of contact with sex organs located at different heights within a flower. Here, we evaluate Darwin's hypothesis in tristylous Lythrum salicaria, a species with a partially tubular corolla, and examine the extent to which the location of sex organs within a flower influence compatible and incompatible pollination. We predicted that the proficiency of cross-pollination would increase for more inserted sex organs due to the restrictions imposed by the floral tube on pollinator positioning. Methods We used experimental trimorphic and monomorphic arrays and emasculated flowers to quantify intermorph pollen transfer and capture among all sex-organ heights, and estimated the contribution of intraflower self-pollination, geitonogamous self-pollination, and intramorph outcross pollination to total intramorph pollination. Key results As predicted, disassortative pollination varied significantly with sex-organ height and was highest for short-level organs and lowest for long-level organs. In monomorphic arrays, most intramorph pollination resulted from outcross pollination followed by intraflower and geitonogamous self-pollination. Conclusions Our results provide evidence for the Darwinian hypothesis. Reciprocal herkogamy promoted varying degrees of disassortative pollination with the magnitude strongly influenced by sex-organ height within a flower.

Most heterostylous plants possess a reciprocal arrangement of stigmas and anthers (reciprocal herkogamy), heteromorphic self-incompatibility, and ancillary polymorphisms of pollen and stigmas. The topographical complementarity hypothesis proposes that ancillary polymorphisms function in the rejection of incompatible pollen thus promoting disassortative pollination. Here, we test this hypothesis by investigating patterns of pollen transfer and capture in populations of dimorphic Armeria maritima and A. pubigera and distylous Limonium vulgare (Plumbaginaceae), and by studying pollen adherence and germination patterns in A. maritima following controlled hand-pollinations. Armeria lacks reciprocal herkogamy allowing the evaluation of the extent to which ancillary polymorphisms affect the composition of pollen loads. We compared the amounts of compatible and incompatible pollen on stigmas in natural populations and calculated the proficiencies of pollen transfer for each mating type. We detected disassortative pollination in each species, and mating types did not differ in compatible pollen capture, although cob stigmas captured more incompatible pollen. Controlled hand-pollinations revealed the failure of incompatible pollen to adhere and germinate on stigmas. Our results provided evidence that, while structural in nature, pollen-stigma dimorphisms are tightly associated with heteromorphic incompatibility and likely function to promote disassortative pollination, especially in the absence of reciprocal herkogamy.

Premise of research. During biological invasion, the genetic diversity of populations may be reduced by founder events and genetic drift. The floral polymorphism tristyly provides an exceptional opportunity to investigate the influence of stochastic forces on the maintenance of genetic polymorphism, because small population size leads to a characteristic signature of morph loss from populations. Here, we investigate the relations between population size and morph-frequency variation in invasive populations of tristylous Lythrum salicaria in Ontario, Canada. We also compare our results to a similar survey conducted 25 years ago in the same region.Methodology. We surveyed the size and morph ratios of 114 L. salicaria populations in 2013. We calculated the relations between population size and style morph absence, population size and style morph evenness, and the number of populations lacking particular style morphs. For comparison of the patterns of morph-frequency variation between surveys, we used a sampl...

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.

Invasion genetics is a relatively new discipline that investigates patterns of genetic variation in populations of invasive species and their ecological and evolutionary consequences. Evolutionary biologists have a long-standing interest in colonizing species, owing to their short life cycles and widespread distributions, but not until publication of The Genetics of Colonizing Species (1965), edited by H.G. Baker and G.L. Stebbins, was a synthesis on the genetics and evolution of colonizers available. Here, I make the case that the Baker and Stebbins volume is the foundational document for invasion genetics, and in conjunction with the increased use of genetic markers and development of invasion biology, resulted in the birth of this new field over the past two decades. I consider the historical origins and legacy of the Baker and Stebbins volume and review some of the key issues that were addressed. I provide biographical sketches of the two editors, emphasizing their contrasting backgrounds and personalities. I review examples from my own work on plant invasions that are relevant to issues discussed by contributors to the volume. These include the following: determinants of invasion success, life history trade-offs, generalist vs. specialist strategies, general-purpose genotypes, adaptive phenotypic plasticity, mating systems and the influence of bottlenecks on genetic variation. I conclude by posing several key questions in invasion genetics and argue that one of the main challenges that the area faces is to integrate experimental field studies of the ecology and demography of populations with the largely descriptive approaches that have tended to dominate most research to date.

Premise of research. The rush family (Juncaceae) is most often described as wind pollinated. However, flowers in the family have pollen in tetrads and numerous ovules, both unusual features for anemophilous plants. Here, we investigate evidence for wind and animal pollination in the alpine rush Juncus allioides in Yunnan Province, southwest China.Methodology. Pollen and ovule traits, insect visitors to flowers, and the relative contribution of wind, insect, and self-pollination to seed set were examined over two years in natural populations.Pivotal results. Unlike most Juncaceae, J. allioides appears to be largely insect pollinated. The pollen/ovule ratio of plants in both populations was ∼700. Few pollen grains were transported by wind to sticky slides in close proximity to plants, and the seed set of bagged-and-emasculated flowers was very low. Although J. allioides is capable of self-fertilization, insect exclusion by bagging reduced seed set in comparison with open pollination. Field observations reve...

Premise of research. Tristyly is a floral polymorphism that is reliably reported from six flowering plant families. Populations of tristylous species often contain three floral morphs that differ reciprocally in stigma and anther height. Trimorphic incompatibility and a suite of ancillary pollen and stigma polymorphisms are commonly associated with sex organ trimorphism. Here, we investigate the tristylous syndrome of Pontederia subovata (Pontederiaceae), a little-known aquatic from southwest Brazil.Methodology. We conducted measurements of sex organ position, pollen size, pollen production, and stigmatic papillae length on the three floral morphs under glasshouse conditions and carried out a controlled pollination program to investigate whether P. subovata possesses a trimorphic incompatibility system and whether its expression varies among the floral morphs.Pivotal results. Pontederia subovata displays reciprocal stigma and anther heights and trimorphism in the size of pollen grains and the length of st...

It is often suggested that traits will be integrated, either because of pleiotropy or because natural selection may favor suites of integrated traits. Plant responses to different environments can provide evidence of such integration. We grew Mercurialis annua plants in high-density stands in high irradiance, in neutral shade, and in high red to far-red (R:FR) shade, resulting in environments of high irradiance, low R:FR; low irradiance, low R:FR; and low irradiance, high R:FR. We measured gas exchange, leaf morphology, stem elongation, and biomass traits and tested the prediction that traits within each functional group would show higher trait integration, as evidenced by high correlations among traits within environments, higher correlations of trait plasticity, and lower plasticity of trait correlations. Overall, we found evidence of only moderate integration for some groups of traits. Functionally related groups of traits, or pairs of traits, could be strongly integrated by one criterion but weakly integrated by another of the criteria. Stem elongation traits, though often observed to be strongly integrated in other taxa, showed little evidence of integration. Internode traits exhibited a novel pattern of responses to low R:FR, with increased elongation of the hypocotyl, decreased elongation of the first internode, and no change in the second internode. We propose that these responses to light are more likely to be the result of natural selection than the consequence of constraints imposed by pleiotropy.

 Premise of the study: The showiness of fl displays is usually explained as an adaptation to attract pollinators. However, selection for less attractive displays imposed by non-pollinating agents, particularly herbivores, may balance pollinator-driven selection for highly visible infl orescences. We investigated whether infl orescence architecture, particularly the unusual groundlevel fl owering associated with a specialized bird perch in Babiana ringens may have originated, in part, as an adaptive response to mammalian herbivory.  Methods: We measured levels of herbivory by antelope in populations of B. hirsuta , the putative sister species of B. ringens , which possesses the likely ancestral form of infl orescence architecture. To test for position-dependent effects of herbivory on fl owers, we compared the herbivory rates and seed production of manipulated infl orescences in a fi eld experiment. We predicted that fl owers at the base of infl orescences would suffer less herbivory than those in apical positions.  Results: We found herbivore damage to flin 50% of naturally occurring B. hirsuta plants. Manipulated infl orescences with only basal fl owers, and consequently similar infl orescence architecture to B. ringens , experienced signifi cantly lower herbivory and higher seed set than infl orescences manipulated to have only apical fl owers.  Conclusions: Our results are consistent with the hypothesis that position-dependent herbivory on infl orescences could have played a role in the evolution of infl orescence design. More specifi cally, position-dependent herbivory may have selected for the loss of apical fl owers. Position-dependent herbivory may have contributed toward the evolution of a naked infl orescence axis, a structure that characterizes B. ringens and functions as a bird perch facilitating cross-pollination by sunbirds.

Variation in pollination intensity can influence the degree of gametophytic competition with diverse consequences for plant fitness. Such effects should be particularly important in wind-pollinated plants because they commonly possess uniovulate flowers. In several dioecious species, competition between female- and male-determining microgametophytes has been proposed as a cause of female-biased sex ratios. Here, we examine functional relations among pollination intensity, seed set, sex ratios, and sporophyte vigor in Rumex hastatulus, a wind-pollinated, dioecious plant. Using controlled pollinations, we applied a range of pollen grain numbers (1–71) to female flowers. Pollination intensity significantly predicted percent seed set, which was >80% when more than 15 pollen grains were added to stigmas. We estimated that the proportion of pollen grains participating in fertilization was 0.28 and the proportion of fertilized ovules developing into seed was 0.89. Overall, progeny sex ratios were female biased (...

Variation in the timing of reproductive functions in dioecious organisms may result in adaptive changes in the direction of sexual dimorphism during the breeding season. For plants in which both pollen and seeds are wind-dispersed, it may be advantageous for male plants to be taller when pollen is dispersed and female plants to be taller when seeds are dispersed. We examined the dynamics of height dimorphism in Rumex hastatulus , an annual, wind-pollinated, dioecious plant from the southern USA. A field survey of seven populations indicated that females were significantly taller than males at seed maturity. However, a glasshouse experiment revealed a more complex pattern of height growth during the life cycle. No dimorphism was evident prior to reproduction for six of seven populations, but at flowering, males were significantly taller than females in all populations. This pattern was reversed at reproductive maturity, consistent with field observations. Males flowered later than females and the degree of height dimorphism was greater in populations with a later onset of male flowering. We discuss the potential adaptive significance of temporal changes in height dimorphism for pollen and seed dispersal, and how this may be optimized for the contrasting reproductive functions of the sexes.

In animal-pollinated plants, reproductive success is commonly limited by pollen availability, which can occur in environments where pollinator activity is scarce or variable. Extended floral longevity to maximize a plant’s access to pollinators may be an adaptation to such uncertain pollination environments. Here, we investigated the effects of flower exposure time to pollinators on female fertility (fruit and seed set) in the bee-pollinated woodland herb Trillium grandiflorum, a species with long-lived flowers (~17-21 d) that blooms in early spring when pollinator activity is often variable. We experimentally exposed flowers to pollinators for different amounts of time to determine the extent to which floral longevity influenced reproductive success. The amount of time that flowers were exposed to pollinators significantly increased fruit set and seed set per flower, but not seed set per fruit. Our results provide experimental evidence that long floral life spans may function as a ‘sit-and-wait’ pollination strategy to increase the amount of exposure time to pollinators and promote seed set in the unpredictable pollination environments often experienced by early spring ephemerals. In large populations with infrequent pollinator visitation, as commonly occurs in T. grandiflorum, pollination may be a largely stochastic process.

Background and aims Dimorphism among floral traits can evolve through variation in selection intensity between female and male performance, especially when sex functions are separated between flowers on a plant (monoecy), or between individuals (dioecy). In animal-pollinated species, male floral traits are predicted to be larger because competition for pollinators should favour larger displays. Floral dimorphism may be greater in dioecious than monoecious populations because of trade-offs between female and male function and opportunities for selfing in hermaphrodites. Methods These predictions were tested by surveying flower size, total flowers per inflorescence and daily display size in the insect-pollinated Sagittaria latifolia (Alismataceae). This species is useful for comparative analysis because populations are mostly either monoecious or dioecious. We examined floral dimorphism in 13 monoecious and 16 dioecious populations in eastern North America. Key results Male flowers were significantly larger than female flowers in monoecious and dioecious populations, but there was no evidence for greater flower size dimorphism in dioecious populations despite their larger flower sizes overall. Although inflorescences in both dioecious and monoecious populations produced more male flowers, daily floral displays were significantly larger for female than male function due to more synchronous female flower opening. Daily floral display dimorphism was significantly greater in dioecious populations, due to greater female daily floral displays. There was a positive relationship between mean flower size and total flowers per inflorescence for both sexes in dioecious populations, but no relationship for either sex function in monoecious populations. Flower size dimorphism was positively correlated with the frequencies of females in dioecious populations. Conclusions The increased size and number of male flowers and protracted male floral displays in S. latifolia are probably shaped by sexual selection for more effective pollen dispersal.

Summary 1. A major goal of DNA barcoding is to identify species in local floras and ecological communities. With the consensus of a two-locus DNA barcode (rbcL+matK) by the Consortium for the Barcode of Life (CBOL) Plant Working Group (2009), barcoding efforts have begun to focus on building the barcode library for land plants. 2. Here, we establish a barcoding database for a temperate flora of moderate taxonomic breadth at the Koffler Scientific Reserve, Ontario, Canada based on the rbcL+matK barcode. We evaluated the performance of this combination in comparison with three other potential supplementary regions (the coding region rpoC1 and two non-coding intergenic spacers trnH-psbA and atpF-atpH). We examined these markers singly and in combination to evaluate their discriminatory power among 436 species in 269 genera of land plants. 3. Using high-throughput techniques, we recovered a high-quality sequence from at least one region for 98.2% of the 513 samples screened; 55% had complete coverage across all five gene regions. Sequencing success was highest for rbcL (91.4% of samples collected) and lowest for rpoC1 (74.5%). The two coding regions rbcL and matK provided a relatively high number of high-quality bi-directional sequences compared with the non-coding intergenic spacers, and in combination were able to correctly identify 93.1% of the species sampled. Marginal increases in species resolution were obtained with the inclusion of the trnH-psbA intergenic spacer (95.3%), or by using all five gene regions combined (97.3%). 4. There was a weak relation between the number of species per genus and identification success rate using rbcL+matK; 100% for monotypic genera (70.5% of the flora) and 83.6% for polytypic genera. Identification success using the rbcL+matK barcode was higher (100%) for gymnosperms, bryophytes, lycophytes and monilophytes (collectively representing 5% of the flora), compared with angiosperms (92.7%). 5. Our results indicate that the rbcL+matK barcode can provide an acceptably high rate of species resolution in the context of this and other local northern temperate floras. It does so in a cost-effective manner, with relatively modest laboratory effort, and despite the presence of missing data from individual plastid regions in a subset of samples.

Our understanding of the spatial organization of root diversity in plant communities and of the mechanisms of community assembly has been limited by our ability to identify plants based on root tissue, especially in diverse communities. Here, we test the effectivenessoftheplastidgenerbcL,acoreplantDNAbarcodingmarker,forinvestigating spatialpatternsofrootdiversity,andrelateobservedpatternstoabove-ground community structure. We collected 3800 root fragments from four randomly positioned, 1-m-deep soil profiles (two vertical transects per plot), located in an old-field community in southern Ontario, Canada, and extracted and sequenced DNA from 1531 subsampled fragments. We identified species by comparing sequences with a DNA barcode reference library developed previously for the local flora. Nearly 85% of sampled root fragments were successfully sequenced and identified as belonging to 29 plant species or species groups. Root abundance and species richness varied in horizontal space and were negatively correlated with soil depth. The relative abundance of taxa belowground was correlated with their frequency above-ground (r = 0.73, P = 0.0001), but several species detected in root tissue were not observed in above-ground quadrats. Multivariate analyses indicated that diversity was highly structured below-ground, and associated with depth, root morphology, soil chemistry and soil texture, whereas little structure was evident above-ground. Furthermore, analyses of species co-occurrence indicates strong species segregation overall but random co-occurrence among confamilials. Our results provide insights into the role of environmental filtering and competitive interactions in the organization of plant diversity below-ground, and also demonstrate the utility of barcoding for the identification of plant roots.

Theory suggests that the contemporary evolution of local adaptation may increase the rate of biological invasion, yet natural selection has rarely been measured in invasive species. A recently published model predicted that latitudinal variation in the strength of stabilizing selection on two correlated traits—flowering time and size—can result in local adaptation during plant invasion but implicitly assumed that population × environment interactions are weak. We tested for stabilizing selection and the effect of growing environment on 13 populations of the invasive plant Lythrum salicaria sampled along a latitudinal gradient. Plants were grown under uniform glasshouse conditions and in a 3-year common garden field experiment at midlatitude. Consistent with model assumptions, the ranking of population means was highly correlated across the 3 years, and with glasshouse measurements, despite significant interannual differences in flowering time and vegetative size. As predicted, the first principal componen...

Summary • Flowering plants display extraordinary diversity in the morphology of male sexual organs, yet the functional significance of this variation is not well understood. Here, we conducted a comparative analysis of floral correlates of heteranthery ‐ the morphological and functional differentiation of anthers within flowers ‐ among angiosperm families to identify traits associated with this condition. • We performed a phylogenetic analysis of correlated evolution between heteranthery and several floral traits commonly reported from heterantherous taxa. In addition, we quantified the effect of phylogenetic uncertainty in the observed patterns of correlated evolution by comparing trees in which polytomous branches were randomly resolved. • Heteranthery is reported from 12 angiosperm orders and is phylogenetically associated with the absence of floral nectaries, buzz-pollination and enantiostyly (mirror-image flowers). These associations are robust to particularities of the underlying phylogenetic hypothesis. • Heteranthery has probably evolved as a result of pollinator-mediated selection and appears to function to reduce the conflict of relying on pollen both as food to attract pollinators and as the agent of male gamete transfer. The relative scarcity of heteranthery among angiosperm families suggests that the conditions permitting its evolution are not easily met despite the abundance of pollen-collecting bees and nectarless flowers.

Colonization may favor self‐compatibility (SC) in invasive plants, a process consistent with Baker’s law. We investigated this hypothesis in invasive eastern North American populations of tristylous Lythrum salicaria L. (purple loosestrife) by controlled self‐ and cross‐pollinations of 124 plants sampled from 12 populations grown under uniform glasshouse conditions. We evaluated whether populations at the northern front of the invasion exhibited higher levels of SC than southern populations, which are closer to source populations for the North American invasion. We also sought evidence for morph‐specific differences in the strength of trimorphic incompatibility. We used the ASTER statistical program to jointly model three measures of SC: (1) the probability of producing at least one seed following self‐pollination, (2) the number of fruits produced by self‐pollination, and (3) the average number of seeds per fruit resulting from self‐pollination. Just over one‐quarter of plants set at least one fruit afte...

In animal-pollinated plants with unisexual flowers, sexual dimorphism in floral traits may be the consequence of pollinator-mediated selection. Experimental investigations of the effects of variation in flower size and floral display on pollinator visitation can provide insights into the evolution of floral dimorphism in dioecious plants. Here, we investigated pollinator responses to experimental arrays of dioecious Sagittaria latifolia in which we manipulated floral display and flower size. We also examined whether there were changes in pollinator visitation with increasing dimorphism in flower size. In S. latifolia, males have larger flowers and smaller floral displays than females. Visitation by pollinators, mainly flies and bees, was more frequent for male than for female inflorescences and increased with increasing flower size, regardless of sex. The number of insect visits per flower decreased with increasing floral display in males but remained constant in females. Greater sexual dimorphism in flower size increased visits to male inflorescences but had no influence on the number of visits to female inflorescences. These results suggest that larger flower sizes would be advantageous to both females and males, and no evidence was found that females suffer from increased flower-size dimorphism. Small daily floral displays may benefit males by allowing extended flowering periods and greater opportunities for effective pollen dispersal.

Wind pollination is predominantly a derived condition in angiosperms and is thought to evolve in response to ecological conditions that render animal pollination less advantageous. However, the specific ecological and evolutionary mechanisms responsible for transitions from animal to wind pollination are poorly understood in comparison with other major reproductive transitions in angiosperms, including the evolution of selfing from outcrossing and dioecy from hermaphroditism. To investigate correlations between wind pollination and a range of characters including habitat type, sexual system, floral display size, floral showiness, and ovule number, we used a large-scale molecular phylogeny of the angiosperms and maximum likelihood methods to infer historical patterns of evolution. This approach enabled us to detect correlated evolution and the order of trait acquisition between pollination mode and each of nine characters. Log likelihood ratio tests supported a model of correlated evolution for wind pollination and habitat type, floral sexuality, sexual system, flower size, flower showiness, presence versus absence of nectar, and ovule number. In contrast, wind pollination and geographical distribution and number of flowers per inflorescence evolve independently. We found that in wind-pollinated taxa, nectar is lost more often and ovule number is reduced to one. We also found that wind pollination evolves more frequently in lineages already possessing unisexual flowers and/or unisexual plants. An understanding of the ecological and life-history context in which wind pollination originates is fundamental to further investigation of the microevolutionary forces causing transitions from animal to wind pollination.

Evolutionary transitions are functionally significant changes in organismal traits that largely result from the action of natural selection. They first appear within populations when novel traits replace the ancestral state because of increased fitness. Evolutionary transitions take on broader significance and can be considered major when newly established traits persist, are maintained through multiple speciation events, and ultimately become well-established features of lineages. The shifts in character state that constitute an evolutionary transition are key elements of biological diversification, and the identification and study of major transitions during the history of life now represent an important research program in evolutionary biology

Using phylogenies to make sound inferences about character evolution depends on a variety of factors, including tree uncertainty, taxon sampling, and the degree of evolutionary lability in the character of interest. We explore the effect of these and other sources of ambiguity for maximum likelihood (ML)–based inferences of sexual‐system evolution in Wurmbea, a small genus of geophytic monocots from the Southern Hemisphere. We reconstructed Wurmbea phylogeny using four noncontiguous regions (ca. 5.5 kb) of the plastid genome across a broad sampling of taxa, and we confirm that the genus is divided into two well‐supported clades, each defined by its geography (Africa vs. Australasia) and variation in sexual system (i.e., uniformly monomorphic vs. sexually variable, respectively). We demonstrate that the predominantly Australian clade includes the sexually monomorphic species Iphigenia novae‐zelandiae. We observe treewide uncertainty in the state of all ancestral nodes, and therefore all state transitions, ...

Enantiostyly, the reciprocal deflection of the style to the left or right side of the floral axis has evolved independently in at least a dozen angiosperm families. Unlike other plant sexual polymorphisms, the adaptive significance of these mirror-image flowers remains unclear. Most authors have interpreted enantiostyly as a floral mechanism that promotes cross-pollination. However, any functional interpretation is complicated by the fact that enantiostyly occurs in two forms. In monomorphic enantiostyly there are left- and right-styled flowers on the same plant, while in dimorphic enantiostyly they are on separate individuals. In this paper we develop a model of pollen transfer which indicates that monomorphic enantiostyly should reduce geitonogamous pollination compared to a non-enantiostylous condition, and that the lowest levels of geitonogamous pollination should occur in dimorphic enantiostyly. We tested these predictions using floral manipulations of bee-pollinated Solanum rostratum in garden arrays. We compared mating patterns and fertility in five array types: non-enantiostylous and straight-styled, monomorphic enantiostylous, dimorphic enantiostylous, and arrays uniform for either left or right stylar deflection. Outcrossing rates in non-enantiostylous arrays (t = 0.33 ± 0.04) were significantly lower than all other arrays, while monomorphic enantiostylous arrays (t = 0.74 ± 0.06) had significantly lower outcrossing rates than dimorphic enantiostylous arrays (t = 0.88 ± 0.04) and those uniform for stylar deflection (t = 0.84 ± 0.04). In dimorphic enantiostylous arrays, intermorph pollen transfer accounted for 75% of all outcrossing events. In pollen-limited situations, both types of enantiostylous arrays had significantly higher female fertility than arrays fixed for one direction, demonstrating that enantiostyly promotes pollen transfer between flowers of opposite stylar orientation. Our results provide support for the hypothesis that enantiostyly functions to increase the precision of cross-pollination in bee-pollinated plants by reducing geitonogamy. © 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 85, 167–179.

We investigated the origin of stylar polymorphisms in Narcissus, which possesses a remarkable range of stylar conditions and diverse types of floral morphology and pollination biology. Reconstruction of evolutionary change was complicated by incomplete resolution of trees inferred from two rapidly evolving chloroplast regions, but we bracketed reconstructions expected on the fully resolved plastid-based tree by considering all possible resolutions of polytomies on the shortest trees. Stigma-height dimorphism likely arose on several occasions in Narcissus and persisted across multiple speciation events. As proposed in published models, this rare type of stylar polymorphism is ancestral to distyly. While there is no evidence in Narcissus that dimorphism preceded tristyly, a rapid transition between them may explain the lack of a phylogenetic footprint for this evolutionary sequence. The single instances of distyly and tristyly in Narcissus albimarginatus and N. triandrus, respectively, are clearly not homologous, an evolutionary convergence unique to Amaryllidaceae. Floral morphology was likely an important trigger for the evolution of stylar polymorphisms: Concentrated-changes tests indicate that a long, narrow floral tube may have been associated with the emergence of stigma-height dimorphism and that this type of tube, in combination with a deep corona, likely promoted, or at least was associated with, the parallel origins of heterostyly.

Trillium erectum L. is an insect-pollinated understory herb widespread in forests of eastern North America. Marker gene studies indicate that the species has a mixed mating system, but aspects of population genetic structure have not been investigated. Using 10 allozyme loci, we measured genetic variation within and among 23 populations sampled from throughout the species' range. Overall, T. erectum displayed moderate levels of genetic diversity in comparison with other herbaceous plants. The percentage of loci that were polymorphic was 52%, with average values (±SE) of 1.20 ± 0.02, 0.08 ± 0.01, and 0.13 ± 0.01 for the number of alleles per locus (A), observed heterozygosity (Ho), and expected heterozygosity (He), respectively. There was evidence of inbreeding within populations (Fis = 0.39, 95% CI 0.26–0.55) and significant population differentiation (Fst = 0.16, 0.05–0.24). Analysis of genetic data provided no evidence of isolation by distance, and together with the occurrence of population subdivision, this suggests that there is relatively limited contemporary gene flow among populations. Northern populations of T. erectum tended to have less genetic variability than southern populations, probably as a result of historical factors associated with post glacial migration. Limited opportunities for gene dispersal as a result of low plant densities, the capacity for self-fertilization, and local seed dispersal by ants are likely to be the main factors maintaining contemporary patterns of genetic variation in T. erectum. Key words: allozymes, genetic diversity, gene flow, population genetic structure, Trillium.

We compared the floral biology of female, male, and cosexual plants in sympatric populations of Wurmbea dioica (Colchicaceae) from Western Australia to assess evidence that changes in pollination biology accompanied a transition between monomorphic to dimorphic sexual systems in this species. In W. dioica, sex‐specific differences in floral design and display, the quantity and quality of rewards, and reproductive phenology represent alternative strategies for pollinator attraction and the avoidance of self‐pollination. Unisexual plants had smaller flowers containing less pollen than flowers of cosexuals; males and cosexuals produced nectar with higher sugar content than did females. These patterns were associated with differences in the composition and visitation rates of pollinators. Nectar‐foraging flies visited all three sexes, but pollen‐collecting bees visited only cosexuals. Fly‐pollinated females achieved greater seed fertility relative to their bee‐pollinated cosexual relatives, and males and cose...

Summary 1 The phenotypic plasticity of vegetative traits is a characteristic feature of aquatic plants, promoting survival and growth in the heterogeneous environments typical of wetlands. Less is known about plastic responses of life-history and reproductive traits, particularly patterns of sex allocation. 2 We investigated the plasticity of vegetative and reproductive traits in Sagittaria latifolia , a clonal aquatic plant whose populations are either monoecious or dioecious. Plants of the two sexual systems exhibit divergent life-history characters associated with the disturbed vs. competitive habitats in which monoecious and dioecious populations occur, respectively. We evaluated the prediction that populations of the two sexual systems would have different patterns of phenotypic plasticity because of the contrasting habitats in which they occur. 3 We grew four clonal replicates of 10 genotypes from seven monoecious and five dioecious populations (total = 480 plants) in two fertilizer treatments under glasshouse conditions and measured components of life history, leaf and flower morphology, and sex allocation. 4 The two sexual systems displayed divergent patterns of plasticity for four life-history traits but only flowering time and ramet production showed the expected pattern of greater plasticity in monoecious populations, and the reverse was true for flower production. Fertilization had opposite effects in the two sexual systems for corm production (increased in monoecious populations) and time to flowering (delayed in dioecious populations). 5 Leaf size generally increased due to the addition of fertilizer; however, this increase was substantially greater in dioecious populations. Larger leaf size in dioecious populations was associated with more convex leaves and greater surface area, potentially increasing light capture in the shaded and more competitive habitats in which these populations occur. 6 We found significant plasticity for female sex allocation in monoecious populations, with more female flowers at higher nutrient levels. However, the majority of populations had a significant genetic component to variation in sex allocation and/or significant genotype × environment interactions. These patterns are consistent with monoecy representing a flexible reproductive strategy for regulating mating opportunities in heterogeneous habitats.

Enantiostyly, the deflection of the style to the left or right side of the floral axis, is a sexual polymorphism reported from at least 10 angiosperm families in both monocotyledons and dicotyledons. To investigate variation in the enantiostylous condition, we examined sex‐organ reciprocity and the arrangements of left‐ and right‐styled flowers in 15 enantiostylous species from eight genera in six families. Enantiostyly can be expressed at three distinct levels of structural organization. In “monomorphic enantiostyly,” individual plants produce both floral forms, either mixed within an inflorescence or segregated between left‐ and right‐styled inflorescences. In “dimorphic enantiostyly,” plants are exclusively left‐ or right‐styled and this condition is a genetic polymorphism. Enantiostyly is most commonly associated with the reciprocal deflection of a pollinating anther (“reciprocal enantiostyly”), although there is a continuum in stigma‐anther position, ranging from no reciprocal anther to complete reci...

The existence of monoecious and dioecious populations within plant species is rare. This limits opportunities to investigate the ecological mechanisms responsible for the evolution and maintenance of these contrasting sexual systems. In Sagittaria latifolia, an aquatic flowering plant, monoecious and dioecious populations exist in close geographic proximity but occupy distinct wetland habitats differing in the relative importance of disturbance and competition, respectively. Life-history theory predicts contrasting evolutionary responses to these environmental conditions. We propose that the maintenance of monoecy and dioecy in S. latifolia is governed by ecological selection of divergent life-history strategies in contrasting habitats. Here we evaluate this hypothesis by comparing components of growth and reproduction between monoecious and dioecious populations under four conditions: natural populations, a uniform glasshouse environment, a common garden in which monoecious and dioecious populations and their F1 progeny were compared, and a transplant experiment using shaded and unshaded plots in a freshwater marsh. Plants from dioecious populations were larger in size and produced heavier corms in comparison with monoecious populations. Monoecious populations flowered earlier and produced more flowers, clonal ramets, and corms than dioecious populations. The life-history differences between the sexual systems were shown to have a quantitative genetic basis, with F1 progeny generally exhibiting intermediate trait values. Survival was highest for each sexual system in field plots that most closely resembled the habitats in which monoecious (unshaded) and dioecious (shaded) populations grow. These results demonstrate that monoecious and dioecious populations exhibit contrasting patterns of investment in traits involved with growth and reproduction. Selection for divergent life histories between monoecious and dioecious populations of S. latifolia appears to be the principal mechanism maintaining the integrity of the two sexual systems in areas of geographic overlap.

Enantiostyly is the deflection of the female sex organ either to the left or to the right side of the floral axis, resulting in mirror-image flowers. Two types of enantiostyly occur: in monomorphic enantiostyly, individuals exhibit both flower forms, whereas in dimorphic enantiostyly, the forms occur on separate plants. Monomorphic enantiostyly is known from at least 10 families, whereas dimorphic enantiostyly is reported in only three. Phylogenetic evidence suggests that monomorphic enantiostyly has evolved from a straight-styled ancestor and that dimorphic enantiostyly is derived from monomorphic enantiostyly. Here, we use theoretical models to investigate the role of pollen transfer in influencing these evolutionary transitions. We used numerical calculations to examine the evolution of monomorphic and dimorphic enantiostyly under different conditions of pollen transfer, inbreeding depression, and pollinator visitation. Our results demonstrate that in comparison to a putative straight-styled ancestor, both monomorphic and dimorphic enantiostyly function to reduce geitonogamous pollen transfer with a concomitant increase in pollen export. Our calculations suggest that the first stage in the evolution of monomorphic enantiostyly involves the deflection of the style only, followed by selection for reciprocity in anther position to promote more precise cross-pollination. Constraints associated with the developmental genetics of left-right asymmetries may account for the low number of evolutionary transitions from monomorphic to dimorphic enantiostyly, despite the evolutionary stability of this condition once it arises.

Aquatic plants are well known for their high degree of phenotypic plasticity in vegetative structures, particularly leaves. Less well understood is the extent to which their sexuality can be modified by environmental conditions. Here we investigate gender plasticity in the European clonal monoecious aquatic Sagittaria sagittifolia (Alismataceae) to determine how floral sex ratios may vary with plant size and inflorescence order. We sampled two populations from aquatic habitats in East Anglia, U.K. and measured a range of plant attributes including ramet size and the number of female and male flowers per inflorescence. The two populations exhibited similar patterns of phenotypic gender, despite contrasting patterns of total allocation to female and male flower number. Plants produced male-biased floral sex ratios but female flower number increased from the first to the second inflorescence whereas male flower number decreased. Size-dependent gender modification occurred in both populations, but the patterns of allocation to female flower production differed between the two populations. Our results are consistent with the view that monoecy is a sexual strategy that enables plants to adjust female and male allocation in response to changing environmental conditions.

Stigma-height dimorphism is a sexual polymorphism in which plant populations are composed of two floral morphs that differ significantly in style length but not anther position. The morphs exhibit approach and reverse herkogamy, floral designs that in most species typically occur as monomorphic conditions. We investigated the floral biology of stigma-height dimorphism in the Mediterranean geophyte Narcissus papyraceus (Amaryllidaceae) in an effort to understand the evolutionary forces maintaining stylar polymorphism. Our survey of 66 populations in Spain, Portugal, and Morocco indicated that 56% were dimorphic with the long-styled morph at an average frequency of 0.79. The remaining 44% of populations sampled were monomorphic for the long-styled morph. In dimorphic populations there was a significant positive relation between population size and the frequency of the short-styled morph. Controlled pollinations demonstrated that N. papyraceus is self-sterile with no significant differences in female fertility between intra- and intermorph crosses. Prior self-pollination reduced seed set in flowers that were subsequently cross-pollinated. Estimates of mating patterns using allozyme markers in eight populations indicated that N. papyraceus is largely outcrossing (mean t(m) = 0.81) with no significant differences between monomorphic and dimorphic populations or style morphs. Stigma-height dimorphism in N. papyraceus is maintained in populations by insect-mediated cross-pollination with biased morph ratios and stylar monomorphism likely resulting from the combined influence of the inheritance of the polymorphism, morph-specific differences in assortative mating and founder effects.

Female fertility in flowering plants is commonly observed to be submaximal. The fraction of ovules in a flower that produce seeds is commonly less than 1.00, particularly in outcrossing species. Here we investigate through controlled pollinations factors that may contribute toward low seed:ovule ratios in Trillium grandiflorum, a self‐incompatible woodland herb from eastern North America. Reduced fruit and seed set in open‐pollinated plants in comparison with hand cross‐pollinated plants demonstrated that pollen limitation was a contributing factor to low female fertility. However, seed:ovule ratios of hand cross‐pollinated flowers averaged only 0.66. Experimental pollinations involving the manipulation of pollen age, the number of pollen donors, the timing of pollination, and the application of self‐pollen before cross‐pollen each had no effect on fertility. This indicated that aspects of pollen delivery were not responsible for low seed:ovule ratios and that resources might influence variation in seed:o...

Enantiostyly is a form of directional asymmetry in plants in which the style is deflected away from the main axis of the flower, either to the left or right side. In Wachendorfia (Haemodoraceae), a small genus of insect-pollinated geophytes restricted to the Cape Province of South Africa, populations are usually polymorphic for asymmetry. Here we investigate dimorphic enantiostyly in the four species of Wachendorfia to determine whether variation in their reproductive systems influences the maintenance of this genetic polymorphism. Experimental field pollinations of W. paniculata and W. thyrsiflora indicated higher fertility for cross- than for self-pollinations, whereas in W. brachyandra, these types of pollination produced similar levels of fertility. Outcrossing rates were highest in W. paniculata (t = 0.78-0.98), with W. brachyandra (t = 0.39-0.79) and W. thyrsiflora (t = 0.76) exhibiting mixed mating systems. Outcrossing rates in two populations of W. parviflora varied from mixed mating (t = 0.61) to predominant selfing (t = 0.07). Population style-morph ratios ranged from 1 : 1 in outcrossing W. paniculata to monomorphism in selfing W. parviflora and clonal W. thyrsiflora. In W. brachyandra, a species with delayed selfing, morph ratios were usually biased. The maintenance of enantiostyly in Wachendorfia appears to be strongly influenced by levels of disassortative mating and the balance between sexual and clonal recruitment.

Post-pollination processes governing mating patterns in Trillium, a well-known genus of insect-pollinated woodland herbs, are poorly understood. Mechanisms influencing outcrossing were investigated in T. grandiflorumand T. erectum, two widespread species native to eastern North America. In southern Ontario, Canada, the two species are often sympatric; they flower in early May, and are pollinated by diAerent assemblages of insects. Controlled cross- and selfpollinations and structural observations of pollen germination and pollen tube growth were conducted to determine whether the two species possess a self-incompatibility (SI) system and, if so, the specific site(s) of self-rejection. Controlled pollinations indicated that both species set significantly more seeds from cross-pollination than self-pollination, implicating the action of SI. This was confirmed by structural studies which demonstrated that selfrecognition and rejection reactions occurred on dry-type stigmatic papillae. Observations of pollen hydration revealed that self-rejection was rapid, being initiated within 10 min of pollination and prior to pollen tube emergence. Final selfrejection resulted in failure of pollen tube growth at the base of stigmatic papillae. SI was expressed more weakly in T. erectum and thereby resulted in considerable self-seed set in some individuals. Estimates of outcrossing rates using allozyme markers indicated that T. erectum displayed a mixed-mating system whereas T. grandiflorum was more highly outcrossed. Structural studies of pollen traits indicated that the two species diAered with respect to the size of grains and their aggregation with implications for pollen dispersal and mating. The ecological and evolutionary implications of the variable expression of SI in Trillium are discussed. # 2001 Annals of Botany Company

The evolution and maintenance of combined vs. separate sexes in flowering plants is influenced by both ecological and genetic factors; variation in resources, partic- ularly moisture availability, is thought to play a role in selection for gender dimorphism in some groups. We investigated the density, distribution, biomass allocation, and physi- ology of sympatric monomorphic (cosexual) and dimorphic (female and male) populations of Wurmbea dioica in relation to soil moisture on the Darling Escarpment in southwestern Australia. Populations with monomorphic vs. dimorphic sexual systems segregated into wet vs. dry microsites, respectively, and biomass allocation patterns and physiological traits reflected differences in water availability, despite similarities in total ramet biomass between the sexual systems. Unisexuals flowered earlier at lower density, and they allocated sig- nificantly more biomass below ground to roots and corms than did cosexuals, which al- located more biomass above ground to leaves, stems, and flowers. Females, males, and cosexuals produced similar numbers of flowers per ramet, but unisexuals produced more ramets than cosexuals, increasing the total number of flowers per genet. Contrary to ex- pectation, cosexuals had significantly higher (more positive) leaf carbon isotope ratios and lower leaf nitrogen content than unisexuals, suggesting that cosexuals are more water-use efficient and have lower rates of photosynthesis per unit leaf mass despite their occurrence in wetter microsites. Cosexuals appear to adjust their stomatal behavior to minimize water loss through transpiration while maintaining high investment in leaves and reproductive structures. Unisexuals apparently maximize the acquisition and storage of both water and nitrogen through increased allocation to roots and corms and enhance the uptake of CO2 by keeping stomata more open. These findings indicate that the two sexual systems have different morphological and physiological features associated with local-scale variation in water availability.

The evolution of floral display is thought to be constrained by trade-offs between the size and number of flowers and inflorescences. We grew in the glasshouse 60 maternal families from each of two Brazilian populations of the annual herb, Eichhornia paniculata. We measured flower size, daily flower number, and total flower number per inflorescence, and two indices of module size, leaf area and age at flowering. We also assessed the size and number of inflorescences produced over 6 weeks. All floral traits exhibited significant heritable variation, some of which was due to genetic variation in module size. Genetic (maternal family) correlations between daily and total flower number did not differ from 1.0, indicating that display size (daily flower number) cannot evolve independently from total flower number per inflorescence. Genetic correlations between flower size and daily flower number ranged from negative to positive (r=–0.78 to +0.84), depending on population and inflorescence. Positive correlations occurred when variation in investment per inflorescence was high so that some families produced both larger and more flowers. These correlations became zero when we controlled for variation in module size. Families that flowered later produced fewer, larger inflorescences (r=–0.33, –0.85). These data support theoretical predictions regarding the combined effects of variation in resource acquisition and allocation on traits involved in trade-offs, and they emphasize the hierarchical organization of floral displays. Our results imply that patterns of resource allocation among inflorescences influence evolutionary changes in flower size and number per inflorescence.

Gender in flowering plants is governed by a complex interplay of genetic and environmental factors. The perennial aquatic herb Sagittaria latifolia displays remarkable variation within and among populations in gender expression, from monoecious populations composed of plants with differing numbers of female and male flowers to completely dioecious populations with separate female and male plants. Here we examine the role of environmental factors in modifying gender expression in these two sexual systems by investigating the relation between ramet size and flower number in three sex phenotypes (hermaphrodite, female, and male). In particular, we assess the extent to which hermaphroditic plants have the option of altering the number of female and male flowers to accommodate their specific environmental circumstances. We sampled 12 populations from diverse wetland habitats in southern Ontario, Canada, estimated their phenotypic gender, and examined the relations between ramet size and a range of vegetative and floral traits. On average, plants from dioecious and monoecious populations did not differ in leaf length, a correlate of ramet size. However, in monoecious populations with significant numbers of both male and hermaphroditic ramets, hermaphrodites were larger and produced more flowers than males. This contrasting pattern was also observed when plants were grown under glasshouse conditions. In monoecious populations, variation in ramet size did not affect the production of male flowers, whereas female flower production varied positively with plant size. These relations enabled statistical prediction of the dependence of gender on plant size and the frequency distribution of gender within monoecious populations. These relations also imply that the male phenotype can be determined environmentally, whereas the female phenotype cannot. The size dependence of floral sex ratios in monoecious populations provides novel insights into the likely evolutionary pathway by which dioecy has evolved from monoecy in Sagittaria.

Trade-offs between flower size and number seem likely to influence the evolution of floral display and are an important assumption of several theoretical models. We assessed floral trade-offs by imposing two generations of selection on flower size and number in a greenhouse population of bee-pollinated Eichhornia paniculata. We established a control line and two replicate selection lines of 100 plants each for large flowers (S+), small flowers (S-), and many flowers per inflorescence (N+). We compared realized heritabilities and genetic correlations with estimates based on restricted-maximum-likelihood (REML) analysis of pedigrees. Responses to selection confirmed REML heritability estimates (flower size, h2 = 0.48; daily flower number, h2 = 0.10; total flower number, h2 = 0.23). Differences in nectar, pollen, and ovule production between S+ and S- lines supported an overall divergence in investment per flower. Both realized and REML estimates of the genetic correlation between daily and total flower number were r = 1.0. However, correlated responses to selection were inconsistent in their support of a trade-off. In both S- lines, correlated increases in flower number indicated a genetic correlation of r = -0.6 between flower size and number. In contrast, correlated responses in N+ and S+ lines were not significant, although flower size decreased in one N+ line. In addition, REML estimates of genetic correlations between flower size and number were positive, and did not differ from zero when variation in leaf area and age at first flowering were taken into account. These results likely reflect the combined effects of variation in genes controlling the resources available for flowering and genes with opposing effects on flower size and number. Our results suggest that the short-term evolution of floral display is not necessarily constrained by trade-offs between flower size and number, as is often assumed.

There has been disagreement concerning the nature of sexual polymorphisms in Narcissus, a genus of insect-pollinated geophytes native to the Mediterranean. The existence of trimorphic heterostyly in the genus has recently been confirmed, but the occurrence of distyly remains enigmatic. All sexually dimorphic species previously investigated possess two distinct style lengths but anthers of similar height. Stigma-height dimorphism does not qualify as true distyly because of the absence of a reciprocal correspondence in stigma and anther position in the floral morphs. Such reciprocal herkogamy is generally regarded as the defining feature of heterostyly. Here we report on distyly in N. albimarginatus (section Apodanthae), a rare species confined to a single mountain in northwestern Morocco. A population composed of equal numbers of long- and short-styled plants exhibited reciprocal herkogamy with lower anthers of the long-styled morph and upper anthers of the short-styled morph corresponding in height to stigmas of short- and long-styled plants, respectively. The presence of both stigma-height dimorphism and distyly in Narcissus is of general significance to theoretical models of the evolution of heterostyly.

We conducted a comparative analysis to determine life history and ecological correlates of pollen limitation among 224 species of animal-pollinated flowering plants. To test predictions concerning the occurrence of pollen limitation, species were classified categorically for six life-history traits (self-compatible vs. self-incompatible, autogamous vs. non-autogamous, specialized vs. unspecialized floral morphology, nectariferous vs. nectarless, monocarpic vs. polycarpic, herbaceous vs. woody) and two ecological conditions (presence in open vs. forested habitats and temperate vs. tropical biomes). Pollen limitation of species in contrasting categories was compared using non-parametric tests (TIPs analysis) and phylogenetically-independent contrasts (PICs). The results of TIPs and PICs analyses were generally congruent, although fewer significant contrasts were evident with PICs, probably because of low statistical power. Overall the results suggest that the influence of phylogenetic history on the intensity of pollen limitation was not particularly strong. Nonetheless, significant variation in the degree of pollen limitation was demonstrated among seven angiosperm families suggesting some phylogenetic component to the phenomenon. With both TIPs and PICs, pollen limitation was less intense in self-compatible and autogamous species. TIPs analysis demonstrated that herbaceous, nectariferous, and temperate species were less likely to be pollen-limited, but using PICs this could only be corroborated for those that were self-incompatible. None of the traits were singularly unambiguous predictors of pollen limitation, possibly reflecting the stochastic nature of pollinator service.

Eichhornia azurea (Pontederiaceae) is a mat- forming, clonal aquatic that inhabits lakes, marshes and river systems in many parts of the Neotropics. The spe- cies is tristylous with long-, mid-, and short-styled morphs commonly represented in natural populations. To investigate whether E. azurea possesses a trimorphic in- compatibility system typical of tristylous species, we conducted a controlled pollination experiment on 15 clones representing the three style morphs from a natural population near Rosario, Argentina. Comparisons of fruit and seed set following self-, illegitimate, and legitimate pollinations clearly demonstrated the presence of trimor- phic incompatibility in E. azurea. Self- and illegitimate pollinations produced significantly less fruit and seed than legitimate pollinations in all three style morphs. Pollen from the two anther levels within a flower exhib- ited contrasting compatibility relations in self-pollina- tions. In common with several other tristylous species in Pontederiaceae, the expression of self-incompatibility was weakest in the mid-styled morph and strongest in the short-styled morph. We discuss the ecological and evolutionary significance of the partial expression of trimorphic incompatibility in E. azurea.

An unusual stylar dimorphism occurs in Narcissus, a plant genus of insect-pollinated Mediterranean geophytes. To determine the characteristics of the sexual polymorphism, we investigated floral variation in 46 populations of N. assoanus (section Jonquillae) and 21 populations of N. dubius (section Tazettae) in SW France. Flowers possess two stamen levels in each morph that occupy slightly different positions within the floral tube. In long-styled plants (L-morph), the stigma is located within or slightly above the upper-level stamens, whereas in short-styled plants (S-morph) the stigma is placed well below the lower-level stamens. The stigma-height dimorphism is distinct from heterostyly because the reciprocity of stigma and anther positions in the two style morphs is only weakly developed and there are no differences between the style morphs in pollen size or production. In both species, mean stigma-anther separation is much greater in the S-morph than the L-morph. In N. assoanus, population style-morph ratios vary from isoplethy (1L:1S) to L-biased, whereas in N. dubius they are usually strongly L-biased or occasionally contain only the L-morph. Populations fixed for the S-morph, or with S-biased morph ratios, were not observed. In N. assoanus, style-morph ratios were associated with population size: large continuous populations always exhibited 1:1 morph ratios, whereas smaller, fragmented populations were often L-biased. This pattern was not evident in N. dubius. We argue that biased style-morph ratios largely result from morph-specific differences in assortative mating.