Correlated evolution of floral morphology and mating-type frequencies in a sexually polymorphic plant

In sexually polymorphic species, reproductive morphology governs mating patterns and the character of negative frequency-dependent selection. If local environmental conditions cause sexual morphs to differ between populations, then frequency-dependent selection should create corresponding geographic variation in morph frequencies. We investigate this relation with a model of morph-ratio evolution and analysis of geographic variation in the heterostylous plant Narcissus triandrus. Unlike other tristylous species, N. triandrus possesses both imperfect reciprocity among morphs in sex-organ position and a self-incompatibility system that permits outcrossing within and between morphs. We sampled 137 populations throughout the Iberian Peninsula for floral-morph ratios, and measured floral morphology in 31 populations. Morph ratios exhibited three atypical features: (1) predominance of the long-styled (L) morph; (2) absence of the mid-styled (M) morph from 17.5% of populations; and (3) a negative relation between the frequencies of the L and M morphs among populations. Morph ratios varied geographically, with decreasing frequency of the M morph from the southeast to the northwest of the species' range. Much of this variation accompanied allometric change in the positions of sex organs, especially the mid-level organs, with the M morph declining in frequency and ultimately being lost in large-flowered populations. Using multivariate multiple regression, we demonstrate that variation in floral morphology among populations predicts this geographic variation in morph frequencies. Our theoretical analysis illustrates that patterns of pollen transfer governed by imperfect sex-organ reciprocity can select for unequal equilibrium morph ratios like those observed for N. triandrus. We interpret the L-biased morph ratios and the unusual morphology of N. triandrus as a consequence of its atypical intramorph compatibility system.     

Geographic variation in floral morphology and style-morph ratios in a sexually polymorphic daffodil

Narcissus triandrus is a tristylous daffodil from the Iberian Peninsula that has striking geographical variation in floral morphology and style-morph ratios. Here, we investigate the relation between this variation and ecological factors to understand mechanisms governing morph ratios. We estimated morph ratios in 124 populations throughout the range of N. triandrus and measured 13 morphological traits in 35-78 populations. Sampling of morph ratios conducted over 2-10-yr intervals demonstrated strong temporal stability. Variation in floral and vegetative traits enabled statistical prediction of morph-frequency variation among populations. Latitudinal gradients in precipitation and temperature were correlated with plant and flower size, with larger flowers in northern populations associated with bumblebee visitation and stylar dimorphism. Flowers of the L- and M-morphs differed significantly from the S-morph in several size-related characters, unlike other tristylous species. This pattern and the similarity in anther positions of the L- and M-morphs suggest that the M-morph of N. triandrus originated through genetic modifiers that shortened styles of the L-morph. Our findings support the hypothesis that geographical variation in style-morph ratios is largely governed by climatic gradients in the Iberian Peninsula, which influence the floral morphology and pollination biology of N. triandrus populations.     

Asymmetrical mating patterns and the evolution of biased style-morph ratios in a tristylous daffodil.

Non-random mating in plant populations can be influenced by numerous reproductive and demographic factors, including floral morphology and inter-plant distance. Here, we investigate patterns of outcrossed mating through male function in Narcissus triandrus, a tristylous, bee-pollinated wild daffodil from the Iberian Peninsula, to test pollen transfer models which predict that floral morphology promotes asymmetrical mating and biased morph ratios. Unlike other tristylous species, N. triandrus has an incompatibility system that permits intra-morph mating and long-level rather than mid-level stamens in the L-morph. Incomplete sex-organ reciprocity should result in significant intra-morph mating in the L-morph. We measured mating patterns in two L-biased populations--dimorphic (two style morphs) and trimorphic (three style morphs)--using multilocus genotyping and maximum-likelihood-based paternity analysis. We also examined the spatial distribution of style morphs and neutral markers to investigate the potential consequence of spatially restricted mating on morph ratios. As predicted, we detected significant amounts of intra-morph mating in the L-morph in both populations. Pollen transfer coefficients generally supported predictions based on the Darwinian hypothesis that anthers and stigmas of equivalent level promote pollinator-mediated cross-pollination in heterostylous populations. There was evidence of significant spatial aggregation of both style morphs and neutral markers in populations of N. triandrus, probably as a result of restricted pollen and seed dispersal. Our results provide empirical support for theoretical models of pollen transfer, which indicate that the commonly observed L-biased morph ratios in Narcissus species result from significant intra-morph mating in the L-morph because of its atypical floral morphology.     

Population structure and genetic diversity in tristylous Narcissus triandrus: insights from microsatellite and chloroplast DNA variation

We investigated cpDNA sequence and nuclear microsatellite variation among populations of the wild daffodil Narcissus triandrus to examine the role of historical vs. contemporary forces in shaping population structure, morphological differentiation and sexual-system evolution. This wide-ranging heterostylous species of the Iberian Peninsula is largely composed of two allopatric varieties (vars. cernuus and triandrus), and populations with either stylar trimorphism or dimorphism. Dimorphic populations only occur in var. triandrus, are mainly restricted to the northwestern portion of the species range, and uniformly lack the mid-styled morph (M-morph). Chloroplast DNA (cpDNA) sequence variation revealed strong geographical structuring and evidence for a fragmentation event associated with differentiation of the two varieties. In var. triandrus, population fragmentation, restricted gene flow and isolation-by-distance were also inferred. Significant differences in genetic diversity and population structure between the two varieties likely reflect historical and contemporary differences in demography and gene flow among populations. Discordance between cpDNA markers and both microsatellite and morphological variation indicate that hybridization has occurred between the two varieties at contact zones. There were no differences in genetic diversity or population structure between dimorphic and trimorphic populations, and chloroplast haplotypes were not associated with either sexual system, indicating transitions in morph structure within each maternal lineage. M-morph frequencies were positively correlated with differentiation at microsatellite loci, indicating that the evolutionary processes influencing these neutral markers also influence alleles controlling the style morphs.     

Spatial ecology of mating success in a sexually polymorphic plant

The spatial context of reproduction is of crucial importance to plants because of their sessile habit. Since pollen and seed dispersal is often restricted, mating success is likely to depend on the quantity and quality of mates in local neighbourhoods. Here we use neighbourhood models to investigate the spatial ecology of pollination and mating in Narcissus assoanus, a sexually polymorphic plant with two mating morphs that differ in style length. By mapping individuals in eight populations from southwestern France, we investigated the influence of the density and morph identity of plants at different spatial scales on variation in female fertility. By using inferences on the expected patterns of pollen transfer based on floral morphology, we were able to predict the quantitative relations between local morph ratios and variation in fertility. Our analyses revealed differences in the spatial clustering of morphs and in their response to plant density and morph identity within local neighbourhoods. Mating success in N. assoanus was characterized by both density- and frequency-dependent processes, a condition that may be a general feature of the spatial ecology of plant mating.     

Evolutionary maintenance of stigma-height dimorphism in Narcissus papyraceus (Amaryllidaceae)

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.     

Elevation-related variation in the population characteristics of distylous Primula nivalis affects female fitness and inbreeding depression

The population characteristics of distylous species are highly sensitive to stochastic natural selection pressure.Therefore,populations growing under different environmental conditions may vary in floral morph ratios,potentially affecting female fitness and leading to inbreeding depression.However,the variation in offspring quality among populations as a result of inbreeding depression is poorly understood in distylous species.This study investigates variations in plant density,seed mass,seed viabilityfemale fitness,and post-dispersal inbreeding depression in both sexual morphs(long-styled and shortstyled plants)of the distylous Primula nivalis that were subjected to different pollination treatments along an elevational gradient from 1657 to 2704 m a.s.l.Population characteristics(morph plant density and ratio)and fruit set were significantly affected by sexual morph and elevation.Plant density and fruitset frequencies were lower for short-styled than for long-styled plants at 2704 m a.s.l.The seeds from the cross-pollinated flowers of both morphs were higher in quality than those of self-pollinated flowers.The female fitness of seeds from cross-pollinated flowers of both morphs was higher than that of seeds from open-pollinated and self-pollinated flowers.The female fitness of seeds from long-styled flowers was higher than that of seeds from short-styled flowers at all elevations.Inbreeding depression increased with elevation among plants with short-styled flowers but not among those with long-styled flowers.Variation in the elevation-dependent mating system might influence female fitness and affect inbreeding depression in both floral morphs.In conclusion,the low quality of seeds from short-styled flowers at high elevations might decrease short-styled flower frequency,affecting population characteristics.     

Genotype frequencies associated with incompatibility systems in tristylous plants

A general procedure has been given previously for calculating frequencies of the morphs Long, Mid, and Short in equilibrium populations of tristylous plants. It is now demonstrated that an equilibrium state actually exists at the genotype level if this procedure produces admissible morph frequencies. This result holds in a diploid model, with or without linkage between the two loci involved. It is shown how the genotype frequencies may be determined for any set of mating probabilities. It is also explained how these frequencies may be calculated in a tetraploid model incorporating double reduction. The general theory is applied to a particular situation where the Mid morph is at a selective disadvantage as a seed parent.     

Pollinator visitation in populations of tristylous Eichhornia paniculata in northeastern Brazil

Summary The frequencies of floral morphs in populations of tristylous Eichhornia paniculata often deviate from the theoretical expectation of equality. This variation is associated with the breakdown of tristyly and the evolution of self-fertilization. Differences in morph frequencies could result from selection pressures due to variable levels of insect visitation to populations and contrasting foraging behavior among the floral morphs. We estimated pollinator densities in 16 populations and quantified visitation sequences to morphs in five populations of E. paniculata in northeastern Brazil. Foraging behavior among floral morphs was measured as the frequency of visits to morphs relative to their frequency in the population (preference) and number of flights between inflorescences of the same versus different morphs (constancy). Pollinator density (number/m²/minute) was not correlated with population size, plant density or morph diversity. Pollinator densities varied most among populations of less than 200 plants. Whether pollinators discriminated among the morphs, depended on whether they primarily collected nectar or pollen. In four populations, nectar-feeding bees (Ancyloscelis and Florilegus spp.) and butterflies showed no consistent preference or constancy among the morphs. In contrast, pollen-collecting bees (Trigona sp.) visited a lower proportion of longstyled inflorescences than expected and tended to visit more mid-and short-styled inflorescences in succession, once they were encountered. Pollinator constancy for morphs did not result from differences in inflorescence production or spatial patchiness among the morphs. Although non-random pollinator visitation to morphs in heterostylous populations could potentially affect mating and hence morph frequencies, the observed visitation patterns in this study do not provide evidence that pollinators play a major role in influencing floral morph frequencies.     

Genetic control of floral morph in tristylous Pickerelweed (Pontederia cordata L.)

Pickerelweed (Pontederia cordata L.) is a diploid (2n = 2x = 16) tristylous aquatic perennial. Populations usually contain 3 floral morphs that differ reciprocally in style length and anther height (referred to as the long-, mid-, and short-styled morphs, hereafter L-, M-, and S-morphs). The floral polymorphism promotes disassortative mating among the 3 floral morphs and is maintained in populations by negative frequency-dependent selection. The objective of this study was to determine the number of loci, number of alleles, and gene action controlling floral morph in pickerelweed. Three parental lines (one each of the L-, M-, and S-morph) were used to create S1 and F1 populations. F2 populations were produced through self-pollination of F1 plants. Progeny ratios of S1, F1, and F2 generations revealed that tristyly is controlled by 2 diallelic loci (S and M) with dominant gene action. The S locus is epistatic to the M locus, with the S-morph produced by plants with the dominant S allele (genotype S _ _ _). Plants with recessive alleles at the S locus were either L-morph (ssmm) or M-morph (ssM_). The results of this experiment demonstrate that the inheritance of tristyly in pickerelweed is the same as previously reported for several tristylous species in the Lythraceae and Oxalidaceae.     

Morph-ratio variation, population size and female reproductive success in distylous Pulmonaria officinalis (Boraginaceae)

Theory predicts that morph ratios in heterostylous populations are governed by negative frequency-dependent selection typically resulting in equal morph ratios at equilibrium. Previous work on the distylous perennial herb Pulmonaria officinalis, however, showed asymmetric mating between floral morphs and a weak self-incompatibility system, with the long-styled morph (L-morph) producing significantly higher seed set following intramorph crosses and even selfing than the short-styled morph (S-morph), two aspects thought to affect female fecundity and morph-ratio variation. Here, we evaluated morph ratios and population size of all known P. officinalis populations in the northern part of Belgium. Morph ratios deviated significantly from 1:1 (range 0.09-1 L-morph frequency, mean = 0.58). Relative fecundity of the S-morph (i.e. mean seed set of the S-morph/mean seed set of the L-morph) was on average 0.73, was positively related to the frequency of the L-morph, and reached 1 (similar levels of female fecundity) at an average L-morph frequency of 0.66 in the population. As some small populations had the S-morph in majority, our results suggest that local morph ratios are influenced both by the relative fecundity of L- and S-morph individuals and by stochastic processes in small populations.     

VARIATION IN MALE AND FEMALE REPRODUCTIVE SUCCESS AMONG FLORAL MORPHS IN THE TRISTYLOUS PLANT LYTHRUM SALICARIA (LYTHRACEAE)

The purpose of this study was to assess variation in male and female reproductive success among the three morphs of the tristylous plant, Lythrum salicaria. Fluorescent dyes were used as pollen analogs to determine whether morphs differ in their abilities to donate and receive pollen, and actual and potential seed set was measured with a hand pollination experiment. Dye transfer among morphs was highly asymmetric, with more frequent transfer from the short-styled morph to the long- and mid-styled morphs. This suggests that shorts are performing better at pollen donation and longs and mids are performing better at pollen receipt. All flowers on 95 plants were hand pollinated to test whether female reproductive success is more pollen-limited in the short-styled morph than in other morphs. Hand-pollinated short-styled plants had significantly higher total seed mass and more seeds per capsule than short controls, whereas hand pollination failed to increase seed set in long and mid morphs. As predicted, short-styled morphs showed significant pollen limitation, whereas seed set in long- and midstyled morphs was not pollen-limited. Thus, in Lythrum salicaria asymmetrical pollen flow generates morph-specific differences in male and female fitness.     

POPULATION STRUCTURE AND MORPH-SPECIFIC FITNESS DIFFERENCES IN TRISTYLOUS LYTHRUM SALICARIA

In tristylous plant populations, style-morph frequencies are governed by an interaction between frequency-dependent selection due to disassortative mating and stochastic processes. Provided that there are no inherent fitness differences among morphs, frequency-dependent selection should result in equal morph frequencies at equilibrium. Stochastic models indicate that the short-styled morph has the highest and the long-styled morph the lowest probability of being lost from local populations as a result of random processes. We surveyed the morph composition of 82 populations of the tristylous, self-incompatible herb Lythrum salicaria in two archipelagos, one in central and one in northern Sweden, located close to the range-margin of the species. To examine whether deviations from even morph frequencies can be explained by among-morph differences in reproductive success, we quantified flower and seed production in six and three populations in the northern and southern archipelago, respectively, and we recorded segregation ratios in offspring produced in six trimorphic populations in the northern area. Seed germination and offspring growth were studied in the greenhouse. Ninety percent of the populations in the southern archipelago (N = 31) and 69% of the populations in the northern archipelago (N = 35) were trimorphic; the remaining populations were dimorphic (only populations consisting of at least three flowering plants considered). Dimorphic populations were smaller than trimorphic populations, as predicted by stochastic models. There was a striking difference in the morph composition of L. salicaria populations between the two archipelagos. In the southern archipelago, there was a slight excess of the long-styled morph and a corresponding deficiency of the short-styled morph. In contrast, the northern populations were characterized by a marked deficiency of the mid-styled morph: the average frequency of the mid-styled morph in trimorphic populations was 0.21, and nine of eleven dimorphic populations lacked the mid-styled morph. In both archipelagos, the long-styled morph (the most common morph) produced about 20% fewer seeds per fruit than the other morphs. The long-styled morph also tended to produce fewer seeds per plant. A hand-pollination experiment performed in two of the northern populations indicated that seed production per flower was pollen-limited in the long-styled morph but not in the other two morphs. Seed germination and offspring size after 24 weeks of growth did not differ among morphs. The mid-styled morph tended to have a higher representation in the offspring than in the parental generation in all six trimorphic populations studied further indicating that the deficiency of the mid-styled morph in the northern archipelago does not represent an equilibrium. Taken together, the results do not support the hypothesis that morph-specific differences in reproductive success can account for deviations from even morph frequencies in L. salicaria. It is suggested that among-morph differences in other components of fitness and historical factors may contribute to the current morph structure.     

THE ROLE OF INBREEDING DEPRESSION AND MATING SYSTEM IN THE EVOLUTION OF HETEROSTYLY

We investigated the role of morph‐based differences in the expression of inbreeding depression in loss of the mid‐styled morph from populations of tristylous Oxalis alpina. The extent of self‐compatibility (SC) of reproductive morphs, the degree of self‐fertilization, and the magnitude of inbreeding depression were investigated in three populations of O. alpina differing in their tristylous incompatibility relationships. All three populations exhibited significant inbreeding depression. In two populations with highly modified tristylous incompatibility, manifested as increased reciprocal compatibility between short‐ and long‐styled morphs, substantial SC and self‐fertilization of mid‐styled morphs were detected, and expected to result in expression of inbreeding depression in the progeny of mid‐styled morphs in the natural populations. In contrast, significant self‐fertility of the mid‐styled morph was absent from the population with typical tristylous incompatibility, and no self‐fertilization could be detected. Although self‐fertilization and expression of inbreeding depression should result in selection against the mid‐styled morph in the later stages of the transition from tristyly to distyly, in O. alpina selection against the mid‐styled morph in the early phases of the evolution of distyly is likely due to genic selection against mid‐alleles associated with modified tristylous incompatibility, rather than expression of inbreeding depression.     

HISTORICAL FACTORS AND ANISOPLETHIC POPULATION STRUCTURE IN TRISTYLOUS PONTEDERIA CORDATA: A REASSESSMENT

Theoretical models of floral-morph frequencies in tristylous species predict a single equilibrium with all three morphs represented in equal proportions (isoplethy). North American populations of Pontederia cordata exhibit considerable heterogeneity of morph frequencies between populations, with the short-styled morph often in excess of isoplethic expectations and the long-styled morph commonly underrepresented. In a previous study, it was proposed that anisoplethic population structure in P. cordata is the result of differential male fertility, owing to genetic differences in pollen production among the morphs. In this study, the influence of historical factors on morph frequencies prior to equilibrium was investigated using a deterministic computer model. Nonequilibrium frequencies are strongly influenced by the genotypes of founding individuals, and, because tristyly is under the control of two diallelic loci, phenotypic equilibrium is approached asymptotically. The model indicates that in nonequilibrium populations the short-styled morph will be in excess and the long-styled morph will be underrepresented. This suggests that historical factors play an important role in determining population structure in P. cordata. Several features of the population ecology of the species lend support to this interpretation. Historical factors should be taken into account when interpreting data from population surveys of morph frequencies in tristylous species and of other genetic polymorphisms not under single-locus control.     

Alternative reproductive strategies and the maintenance of female color polymorphism in damselflies

Genetic polymorphisms are powerful model systems to study the maintenance of diversity in nature. In some systems, polymorphisms are limited to female coloration; these are thought to have arisen as a consequence of reducing male mating harassment, commonly resulting in negative frequency‐dependent selection on female color morphs. One example is the damselfly Ischnura elegans, which shows three female color morphs and strong sexual conflict over mating rates. Here, we present research integrating male tactics, and female evolutionary strategies (female mating behavior and morph‐specific female fecundity) in populations with different morph‐specific mating frequencies, to obtain an understanding of mating rates in nature that goes beyond the mere measure of color frequencies. We found that female morph behavior differed significantly among but not within morphs (i.e., female morph behavior was fixed). In contrast, male tactics were strongly affected by the female morph frequency in the population. Laboratory work comparing morph‐specific female fecundity revealed that androchrome females have lower fecundity than both of the gynochrome female morphs in the short term (3‐days), but over a 10‐day period one of the gynochrome female morphs became more fecund than either of the other morphs. In summary, our study found sex‐specific dynamics in response to different morph frequencies and also highlights the importance of studying morph‐specific fecundities across different time frames to gain a better understanding of the role of alternative reproductive strategies in the maintenance of female‐limited color polymorphism.     

Disassortative mating prevails in style‐dimorphic Narcissus papyraceus despite low reciprocity and compatibility of morphs

Evolution to reduce inbreeding can favor disassortative (intermorph) over assortative (intramorph) mating in hermaphroditic sexually polymorphic plant species. Heterostyly enhances disassortative pollination through reciprocal placement of stigmas and anthers of morphs and appropriate pollinators. Stylar dimorphism in which there is not reciprocal anther placement may compromise disassortative mating, particularly when there is not intramorph incompatibility. Variable rates of disassortative mating along with differential female fecundity or siring success among floral morphs could lead to variation in morph ratio. We investigated mating patterns, female fecundity, and siring success of style‐length morphs in Narcissus papyraceus, a self‐incompatible but morph‐compatible species with dimorphic (long‐ and short‐styled) and monomorphic (long‐styled) populations in central and north regions of its range, respectively. We established experimental populations in both regions and exposed them to ambient pollinators. Using paternity analysis, we found similar siring success of morphs and high disassortative mating in most populations. Female fecundity of morphs was similar in all populations. Although these results could not completely explain the loss of dimorphism in the species’ northern range, they provided evidence for the evolutionary stability of stylar dimorphism in N. papyraceus in at least some populations. Our findings support the hypothesis that prevailing intermorph mating is key for the maintenance of stylar dimorphism.     

THE DISSOLUTION OF A COMPLEX GENETIC POLYMORPHISM: THE EVOLUTION OF SELF-FERTILIZATION IN TRISTYLOUS EICHHORNIA PANICULATA (PONTEDERIACEAE)

Eichhornia paniculata (Pontederiaceae) displays a wide range of outcrossing levels as a result of the dissolution of the tristylous genetic polymorphism and the evolution of semihomostyly. Population surveys, comparison of fitness components of the style morphs, and computer simulations were used to investigate the breakdown of tristyly and the selective mechanisms responsible for the evolution of self-fertilization. Of 110 populations surveyed in northeast Brazil and Jamaica, 53% were trimorphic, 25% were dimorphic, and 22% were monomorphic for style morph. The short (S) morph was underrepresented in trimorphic populations and absent from nontrimorphic populations. The mid (M) morph predominated in dimorphic populations and was the only morph in monomorphic populations. Stamen modifications promoting selfing, associated with semihomostyle evolution, were largely confined to the M morph. They were rare in trimorphic populations, common in dimorphic populations, and often fixed in monomorphic populations. Stochastic simulations and comparisons of fruit set in natural populations indicate that founder events, population bottlenecks, and lowered fertility of the S morph due to an absence of long-tongued pollinators can each account for loss of the S morph from trimorphic populations. A reduced level of disassortative mating can accentuate the rate at which the S morph is lost by both random and deterministic processes. Nontrimorphic populations occur at the geographical margins of the region surveyed and tend to be smaller and less dense than trimorphic populations. These observations and the higher fruit set of the M morph relative to the L morph in dimorphic populations suggest that reproductive assurance, favoring selfing variants of the M morph under conditions of low pollinator service, has been of primary importance in the origin of most monomorphic populations. Where pollinator service is reliable, however, automatic selection of selfing genes, aided by mating asymmetries between the morphs, can cause the M morph to spread to fixation in dimorphic populations.     

Fertility differences among floral morphs following selfing in tristylous Eichhornia paniculata (Pontederiaceae): inbreeding depression or partial incompatibility?

Reduction in seed set following self- vs. cross-pollination in flowering plants can result from abortion of selfed offspring owing to inbreeding depression and/or partial self-incompatibility. Previous studies on tristylous Eichhornia paniculata (Pontederiaceae) indicate that reduced seed set following self-pollination generally occurs in the short- (S), but not the long-(L) or mid-styled (M) morphs. To determine whether this pattern results from morph-specific differences in inbreeding depression owing to the sheltering of deleterious alleles at the S locus and/or partial self-incompatibility, we conducted controlled hand-pollinations of the floral morphs and measured seed set and levels of seed abortion. There were no significant differences in fertilization success and seed set following self-, illegitimate, and legitimate pollinations in the L and M morphs. In contrast, in the S morph self-, intramorph and intermorph illegitimate pollinations resulted in significant reduction in seed set in comparison with legitimate pollination. This indicates that the reduced seed set observed in self-pollination is the result of partial incompatibility rather than inbreeding depression. Significantly reduced fertilization success and low levels of ovule abortion in illegitimate pollinations of S plants also supported this conclusion. Reduced fertility in the S morph may have implications for the observed loss of this morph from natural populations and the evolutionary breakdown of tristyly.     

Frequency-dependent variation in reproductive success in Narcissus: implications for the maintenance of stigma-height dimorphism

Negative frequency-dependent selection is a major selective force maintaining sexual polymorphisms. However, empirical demonstrations of frequency-dependent reproductive success are rare, particularly in plants. We investigate this problem by manipulating the frequencies of style morphs in a natural population of Narcissus assoanus, a self-incompatible herb with style-length dimorphism and intra-morph compatibility. We predicted that the reproductive success of morphs would vary negatively with their frequency because of the effects of morph-specific differences in sex-organ position on patterns of pollen transfer. This prediction was generally supported. The fruit and seed set of the two morphs did not differ significantly in plots with 1 : 1 morph ratios. However, short-styled plants produced significantly fewer seeds than long-styled plants in monomorphic plots, and significantly more seeds than long-styled plants in plots with 'long-biased' morph ratios. These patterns indicate that in the absence of physiological barriers to intra-morph mating, negative frequency-dependent selection contributes to the maintenance of stylar polymorphism through inter-morph pollen transfer. Our experimental results also provide insights into the mechanisms governing the biased style-morph ratios in populations of Narcissus species.