The bees don't know and the flowers don't care: the effect of heterospecific pollen on reproduction in co‐occurring Thysanotus species (Asparagaceae) with similar flowers

Species integrity relies on the maintenance of reproductive isolation, particularly between closely related species. Further, it has been hypothesized that the presence of heterospecific pollen on flower stigmas adversely affects plant reproduction with increasing effect in closely related species. Using two pairs of co‐occurring buzz‐pollinated Thysanotus spp. in the Mediterranean climate region of Perth, Western Australia, we quantified the effect of heterospecific pollen on fruit and seed set. We first determined the mating systems of the two focal species using self‐ and outcross pollen, followed by separate treatments with heterospecific pollen within each species pair. Of the two species receiving pollen, Thysanotus triandrus had a mixed mating system, but with significantly lower fruit and seed set from self‐pollen relative to outcross pollen. Thysanotus tenellus was autogamous with no difference in fruit or seed set between autogamous, self‐ or outcross pollinations. Heterospecific pollen had no effect on fruit or seed set of either focal species. These outcomes point to post‐pollination reproductive isolation, consistent with a floral morphology that causes low specificity of pollen placement and thus a poor capacity for pre‐pollination discrimination. Negative effects of heterospecific pollen, therefore, do not appear to play a role in the reproduction in this group of buzz‐pollinated flowers.     

The breeding system of Lycium cestroides: a Solanaceae with ovarian self-incompatibility

To determine the breeding system of Lycium cestroides,several treatments were performed: self-, cross-, and geitonogamous pollinations, autonomous self-pollination, and a control (flowers exposed to natural pollinators). Production and quality of fruits and seeds as well as pollen tube growth were evaluated for each treatment. Experimental pollinations indicate that L. cestroides is self-incompatible since fruits were obtained only under cross-, and open-pollination treatments. However, in self- and geitonogamous hand pollinations, as well as in autonomous self-pollination, pollen tubes developed successfully and reached the ovules. The speed of pollen tube growth did not differ significantly among the different hand-pollinated flowers (cross-, self-, and geitonogamous). These facts indicate the presence of an ovarian self-incompatibility system. Significant differences were observed in fruit set, fruit size, and seed number per fruit between cross-pollinated flowers and open pollination (control). These results could be explained in terms of quantity and quality of pollination in each case. Received: 2 November 2000 / Revision accepted: 26 January 2001     

Autogamous seed set in a critically endangered orchid in Japan: pollination studies for the conservation of Nervilia nipponica

The pollination biology of the critically endangered Nervilia nipponica was investigated at two sites on Shikoku Island, Japan, to shed light on the species' ecological strategy and to help prioritise management decisions for its conservation. Though the solitary flowers failed to attract pollinators, high rates of fruit set were observed in control plants at both sites. Comparable rates were obtained in bagged plants and following induced autogamous and xenogamous pollinations, indicating that the species is capable of outbreeding, but self-compatible and not pollinator-limited for fruit set under natural conditions; rates were significantly lower following emasculation. The number of seeds set per fruit and their fertility did not differ significantly between open-, self- and cross-pollinated treatments. Epifluorescence microscopy revealed that, in open-pollinated plants, pollen grains germinate within the anther on the first day of anthesis, and that fertilisation occurs within the following 96 hours. The position of the stigma, which overlaps the clinandrium, friable pollinia, and the absence of a rostellum ensure efficient, non-mechanical autopollination. Though this confers reproductive assurance, inbreeding depression may be limiting fitness at subsequent stages of the recruitment cycle and low genetic diversity is inferred.     

Pollen limitation in the endangered Chinese endemic species Sinocalycanthus chinensis

Pollen limitation negatively impacts endangered and endemic plants with small fragmented populations, such as Sinocalycanthus chinensis, an endangered plant endemic to China. In this study, we analyzed the pollen limitation of the S. chinensis Damingshan (DMS) population in 2006, 2009, and 2010, and crossed plants with mates separated by different distances, both within and between populations. The DMS population exhibited strong pollen limitation in fruit set, seed set, and seeds per fruit in 2006, 2009, and 2010. The average accumulated pollen limitation (for fruit set times seeds per fruit) was 0.510 ± 0.180. Progeny crossed with pollen from intermediate neighboring plants within the same population (separated by 30–50 m from pollen recipients) had the lowest fitness. No optimal outcrossing distance was found within the DMS population. Progeny from crosses with the Shunxiwu (SXW) and Daleishan (DLS) populations performed relatively better, while those from crosses with Qingliangfeng (QLF) and Longxushan (LXS) populations performed worse. Compared with average reproductive success, outbreeding depression was found in progeny from crosses with the LXS and QLF populations. Reproductive success from pure self‐pollination indicated S. chinensis is self‐compatible. Geitonogamous selfing increased reproductive success. Based on geitonogamous selfing, the proportion of selfed offspring was relatively high. These results provide basic references for the conservation of this species.     

How geitonogamous selfing affects sex allocation in hermaphrodite plants

Does the mode of self-pollination affect the evolutionarily stable allocation to male vs. female function? We distinguish the following scenarios. (1) An ‘autogamous’ species, in which selfing occurs within the flower prior to opening. The pollen used in selfing is a constant fraction of all pollen grains produced. (2) A species with ‘abiotic pollination’, in which selfing occurs when pollen dispersed in one flower lands on the stigma of a nearby flower on the same plant (geitonogamy). The selfing rate increases with male allocation but a higher selfing rate does not mean a reduced export of pollen. (3) An ‘animal-pollinated’ species with geitonogamous selfing. Here the selfing rate also increases with male allocation, but pollen export to other plants in the population is a decelerating function of the number of simultaneously open flowers. In all three models selfing selects for increased female allocation. For model 3 this contradicts the general opinion that geitonogamous selfing does not affect evolutionarily stable allocations. In all models, the parent benefits more from a female-biased allocation than any other individual in the population. In addition, in models 2 and 3, greater male allocation results in more local mate competition. In model 3 and in model 2 with low levels of inbreeding depression, hermaphroditism is evolutionarily stable. In model 2 with high inbreeding depression, the population converges to a fitness minimum for the relative allocation to male function. In this case the fitness set is bowed inwards, corresponding with accelerating fitness gain curves. If the selfing rate increases with plant size, this is a sufficient condition for size-dependent sex allocation (more allocation towards seeds in large plants) to evolve. We discuss our results in relation to size-dependent sex allocation in plants and in relation to the evolution of dioecy.     

Pollen source affects female reproductive success and early offspring traits in the rare endemic plant Polemonium vanbruntiae (Polemoniaceae)

Understanding the relative magnitudes of inbreeding and outbreeding depression in rare plant populations is increasingly important for effective management strategies. There may be positive and negative effects of crossing individuals in fragmented populations. Conservation strategies may include introducing new genetic material into rare plant populations, which may be beneficial or detrimental based on whether hybrid offspring are of increased or decreased quality. Thus, it is important to determine the effects of pollen source on offspring fitness in rare plants. We established pollen crosses (i.e. geitonogamous‐self, autonomous‐self, intrasite‐outcross, intersite‐outcross and open‐pollinated controls) to determine the effects of pollen source on fitness (seeds/fruit and seed mass) and early offspring traits (probability of germination, number of leaves, leaf area and seedling height) in the rare plant Polemonium vanbruntiae. Open‐pollinated, intrasite‐outcross and geitonogamous‐self treatments did not differ in fitness. However, plants receiving autonomous‐self pollen had the lowest fitness and the lowest probability of seed germination. Intersite‐outcross plants contained fewer seeds/fruit, but seeds germinated at higher frequencies and seedlings were more vigorous. We also detected heterosis at the seed germination stage. These data may imply that natural populations of P. vanbruntiae exhibit low genetic variation and little gene flow. Evidence suggests that deleterious alleles were not responsible for reduced germination; rather environmental factors, dichogamy, herkogamy and/or lack of competition among pollen grains may have caused low germinability in selfed offspring. Although self‐pollination may provide some reproductive assurance in P. vanbruntiae, the result is a reduction in germination and size‐related early traits for selfed offspring.     

Plant size,geitonogamy and seed set in Ipomopsis aggregata

Summary We used powdered fluorescent dyes to estimate receipt of self vs. outcross pollen in the self-incompatible species Ipomopsis aggregata (Polemoniaceae). Flowers on small and large plants received equal amounts of outcross pollen, whereas flowers on large plants received more self pollen, so the proportion of self pollen delivered through geitonogamy increased with plant size. In natural populations emasculation of all flowers on a plant raised average seed set per flower from 5.19 to 6.99 and also raised fruit set, though not significantly. From these results one expects a negative correlation between plant size and seeds per flower. The opposite trend was observed in a sample of plants in the field, suggesting that deleterious effects of geitonogamy on female fecundity in large plants can be overruled by other factors such as size-related fruit or seed abortion. Results are discussed in relation to the evolution of gynodioecy.     

Pollination biology of Harrisia portoricensis (Cactaceae), an endangered Caribbean species

Traits associated with self-pollination are common features of island plant communities. In this work, we studied the pollination biology and the breeding system of Harrisia portoricensis, an island columnar cactus, to test for the presence of inbreeding and inbreeding depression. For H. portoricensis, which bears flowers with typical outcrossing morphology, the results from 322 h of direct observations and videotaping showed that visits to flowers by animals were uncommon. Controlled pollinations demonstrated that H. portoricensis has a partially self-compatible breeding system that it is not autogamous and thus requires an external mechanism for the movement of pollen to set fruit. We detected differences in seed size, seed mass, germination success, and multiplicative fitness estimates between self- and cross-pollination treatments. We found that progeny resulting from natural and self-pollination treatments showed signs of inbreeding depression compared with progeny resulting from cross-pollination; however, the magnitude of the inbreeding depression was less than 50%. Our combined results suggest that for this species an endogamous breeding system should be favored by natural selection.     

Heterostyly promotes compatible pollination in buckwheats: Comparisons of intraflower,intraplant, and interplant pollen flow in distylous and homostylous Fagopyrum

Premise of the Study

Heterostyly, the reciprocal positioning of stigmas and anthers in different floral morphs, has long been thought to promote intermorph pollination. However, extensive intramorph pollination occurs commonly in heterostylous species, leading to recurrent questions about the functional and evolutionary significance of heterostyly.

Methods

To identify the sources of stigmatic pollen (autogamous [intraflower], geitonogamous [intraplant], vs. interplant), we emasculated either one flower or entire plants in experimental populations of the two closely related buckwheat species, distylous Fagopyrum esculentum and homostylous F. tataricum. Differences in pollen size allowed unambiguous identification of pollen on stigmas.

Results

Only 2.4% of F. tataricum pollen and 1.5% of F. esculentum pollen arrived successfully on compatible stigmas of other plants. In the former (homostylous) species, 71.3% of the pollen load on stigmas was autogamous, 10.8% was geitonogamous, and 17.9% was interplant. In the latter (distylous) species, 37.45% of the pollen on stigmas was autogamous, 13.8% was geitonogamous, 17.0% was intramorph, and 31.75% was intermorph. The amount of incompatible pollen arriving on stigmas was greatly decreased by both one‐flower and whole‐plant emasculations, and thus, the proportion of compatible pollen deposited increased with one‐flower emasculation and increased even more with whole‐plant emasculation.

Conclusions

Our quantification of pollen‐donor sources in these two species indicated that heterostyly in Fagopyrum esculentum provided a nearly 2‐fold fitness advantage (in terms of compatible pollination) over expected (random) pollen transfers between morphs. Because of reduced herkogamy, the homostylous F. tataricum was highly autogamous.     

Floral Structure, Stigma Receptivity and Pollen Viability in Relation to Protandry and Self-incompatibility in Silky Oak (Grevillea robusta A. Cunn.)

The reproductive biology of Grevillea robusta growing underexotic conditions in Kenya and Australia is reported. The speciesshowed both protandry and a self-incompatibility mechanism.The stigma was wet and papillate with a distinct groove in themiddle. The anthers dehisced prior to anthesis, when the perianthopened. Stigmatic receptivity began 1 d after anthesis, withthe greatest pollen germination rates and longest pollen tubesobtained 2 d after anthesis. Nectar secretion commenced withpollen dehiscence and was abundant at anthesis. Most stigmaticgrooves opened widely 1–2 d after anthesis and stigmasshowed taller papillae and abundant secretion. Controlled pollinationsgave a greater fruit set from cross-pollination (5.9% in Apriland 17.5% in July) than open-pollination (0.1% in April and3.3% in July). No fruit set from self-pollination was obtainedin April, and very few fruit set for geitonogamous (two outof 1622; 0.1%) or for autogamous (one out of 2707 flowers; 0.04%)pollination treatments in July. Following self-pollination,growth of pollen tubes was poorer than in other treatments,and was generally arrested in the upper style. Cross-pollinatedflowers produced normal and straight pollen tubes, while self-pollentubes had growth abnormalities. Most of the open-pollinatedflowers were found without pollen or with only self-pollen ontheir stigmas indicating that the amount of cross-pollen reachingthe stigma under open-pollination may be a factor limiting seedproduction. Flowers shed soon after the fertilization phasewere those with ungerminated pollen or no pollen. Although avery low rate of selfing may occur, G. robusta presents a self-incompatibilitysystem and allogamy is its primary breeding behaviour.Copyright2000 Annals of Botany Company Grevillea robusta, silky oak, Proteaceae, protandry, controlled pollinations, receptivity, pollen-tube growth, self-incompatibility, pollination     

Genetic consequences of specialization: yucca moth behavior and self-pollination in yuccas

Reciprocal specialization in interspecific interactions, such as plant-pollinator mutualisms, increases the probability that either party can have detrimental effects on the other without the interaction being dissolved. This should be particularly apparent in obligate mutualisms, such as those that exist between yucca and yucca moths. Female moths collect pollen from yucca flowers, oviposit into floral ovaries, and then pollinate those flowers. Yucca moths, which are the sole pollinators of yuccas, impose a cost in the form of seed consumption by the moth larvae. Here we ask whether there also is a genetic cost through selfish moth behavior that may lead to high levels of self fertilization in the yuccas. Historically, it has been assumed that females leave a plant immediately after collecting pollen, but few data are available. Observations of a member of the Tegeticula yuccasella complex on Yucca filamentosa revealed that females remained on the plant and oviposited in 66% of all instances after observed pollen collections, and 51% of all moths were observed to pollinate the same plant as well. Manual cross and self pollinations showed equal development and retention of fruits. Subsequent trials to assess inbreeding depression by measuring seed weight, germination date, growth rate, and plant mass at 5 months revealed significant negative effects on seed weight and germination frequency in selfed progeny arrays. Cumulative inbreeding depression was 0.475, i.e., fitness of selfed seeds was expected to be less than half that of outcrossed seeds. Single and multilocus estimates of outcrossing rates based on allozyme analyses of open-pollinated progeny arrays did not differ from 1.0. The discrepancy between high levels of behavioral self-pollination by the moths and nearly complete outcrossing in mature seeds can be explained through selective foreign pollen use by the females, or, more likely, pollen competition or selective abortion of self-pollinated flowers during early stages of fruit development. Thus, whenever the proportion of pollinated flowers exceeds the proportion that can be matured to ripe fruit based on resource availability, the potential detrimental genetic effects imposed through geitonogamous pollinations can be avoided in the plants. Because self-pollinated flowers have a lower probability of retention, selection should act on female moths to move among plants whenever moth density is high enough to trigger abortion. Received: 18 March 1996 \Accepted: 30 July 1996     

Reproductive versatility in legumes: the case of amphicarpy in Trifolium polymorphum

Amphicarpy is a fascinating reproductive strategy, defined as fruit produced both below the soil surface and as aerial fruit on the same plant. Trifolium polymorphum is a grassland species subject to herbivory that combines amphicarpy with vegetative reproduction through stolons. Underground flowers have been described as obligate autogamous and aerial ones as self‐compatible allogamous, with aerial floral traits favouring cross‐pollination. In the present work we performed different pollination treatments on aerial flowers to analyse rates of pollen tube development and offspring fitness, measured as fruit set, seed production and germination percentage. This last variable was compared to that of seeds produced underground. No significant differences were found between fruit set in self‐ and cross‐pollinations. Seed production was higher in self‐pollinations, which is consistent with the higher rate of pollen tube development observed in self‐crosses. Spontaneous self‐pollination is limited in aerial flowers; thus pollen transfer by means of a vector is required even within the same flower. Germination tests showed that aerial seeds produced after self‐ and cross‐pollination did not differ in fitness, but underground seeds had higher germination percentage than aerial ones. Thus, we conclude that T. polymorphum has a mixed mating system. In grasslands with heavy grazing pressure, clonal propagation and underground seed production ensure persistence in the field. An intermediate level of selfing in aerial flowers ensures offspring, but morphological (herkogamy) and functional (dicogamy) floral traits maintain a window to incorporate genetic variability, allowing the species to tolerate temporal and spatial pressures.     

Dissecting components of flowering pattern: size effects on female fitness

Flowering synchrony is essential for plant reproductive success, especially in the case of small‐sized populations of self‐incompatible species. Closely related to synchrony, flowering intensity influences pollinator attraction and pollinator movements. Thus, a high flowering intensity may increase pollinator attraction but, at the same time, may also increase the probability of geitonogamous pollinations. Depending on the mating system, the female fitness of plants in small populations may be affected by both the positive effects of higher flowering synchrony and pollinator attraction and the negative effects of geitonogamous pollinations induced by a high flowering intensity. It was hypothesized that different‐sized plants in a population would show contrasting flowering patterns, resulting in differences in pollinator behaviour. These influences could result in differences in mating and female reproductive success. This hypothesis was tested by studying the flowering pattern of Erodium paularense (Geraniaceae), a rare and endangered endemic of central Spain. The temporal distribution of flower production was explored throughout the reproductive season, and the probability of xenogamy and geitonogamy and their relationship to plant size and fitness components were calculated. The analysis of this partially self‐compatible species showed diverse flowering patterns related to different plant sizes. Small plants produced a larger number of seeds per fruit in spite of having lower values of flowering synchrony. By contrast, large plants produced a larger number of seeds from geitonogamous pollinations. The effect of different flower displays and outcrossing rates on seed set varied throughout the season in the different groups. Our findings highlight the relevance of individual plant size‐dependent phenology on female reproductive success and, in particular, on the relationship between flowering synchrony and fitness. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 156 , 227–236.     

REPRODUCTIVE STRATEGY OF CLEISTES DIVARICATA (ORCHID ACEAE)

In Cleistes divaricata, a hinged anther dispenses a sequence of loosely aggregated pollen tetrad masses onto the dorsal thorax of Bombus and Megachile workers. This mechanism allows more than one bee to receive pollen from a single flower. To determine the effectiveness of this strategy, plants in bud were caged and hand pollinated using pollen from sequential releases. Capsules produced from a flower's first three pollen dumps were significantly larger and heavier and contained more seeds than those produced by later pollen deposits with fewer tetrads. Decreasing pollen dosage did not affect fruit set, capsule development, individual seed weight, or percentage of healthy looking embryos per fruit. Fruit set for artificial pollinations was 93% for pollen dumps 1–3 and 97% for dumps 4–18. As set of naturally pollinated marked plants was only 38%, low frequency of pollinator visits apparently limited seed production. Efficacy of infrequent visits is maximized, however, because a flower's first pollen releases contain the most pollen, but later low-dosage dumps can produce some seeds. An unexpected cost was a higher investment of pericarp per seed in low-dosage capsules (130 ng pericarp seed^-1 fresh weight) than in high-dosage pollinations (only 27 ng pericarp seed^–1). Flower fading in plants receiving low pollen dosage was as slow as that in caged, unpollinated controls, thus increasing opportunity for additional pollination.     

Pollination ecology and inbreeding depression control individual flowering phenologies and mixed mating

We analyze evolution of individual flowering phenologies by combining an ecological model of pollinator behavior with a genetic model of inbreeding depression for plant viability. The flowering phenology of a plant genotype determines its expected daily floral display which, together with pollinator behavior, governs the population rate of geitonogamous selfing (fertilization among flowers on the same plant). Pollinators select plant phenologies in two ways: they are more likely to visit plants displaying more flowers per day, and they influence geitonogamous selfing and consequent inbreeding depression via their abundance, foraging behavior, and pollen carry‐over among flowers on a plant. Our model predicts two types of equilibria at stable intermediate selfing rates for a wide range of pollinator behaviors and pollen transfer parameters. Edge equilibria occur at maximal or minimal selfing rates and are constrained by pollinators. Internal equilibria occur between edge equilibria and are determined by a trade‐off between pollinator attraction to large floral displays and avoidance of inbreeding depression due to selfing. We conclude that unavoidable geitonogamous selfing generated by pollinator behavior can contribute to the common occurrence of stable mixed mating in plants.     

SELF-INCOMPATIBILITY SYSTEMS IN THE RUBIACEAE OF A TROPICAL LOWLAND WET FOREST

A program of experimental field pollinations was carried out on 14 species of woody Rubiaceae in a Costa Rican wet forest in order to test for the presence of self-incompatibility systems. Species of Cephaelis, Coussarea, Faramea, Hamelia, Posoqueria, Psychotria, Rudgea, and Warszewiczia were investigated in the study. Ten of the species are distylous, and of these, nine were found to be self-incompatible. The site of the incompatibility barrier varied between and within species. Short style plants always had stigmatic inhibition of pollen tubes following self and intraform pollinations, but the site of the rejection response in long style plants was quite variable. In several species, pollen tubes, resulting from incompatible pollinations of the long style flowers, penetrated to the base of the style. Fruit set was followed in two of these species, and despite deep penetration of self and intraform pollen tubes in the long style morph, only interform pollinations resulted in fruit set. Four of the species tested are florally monomorphic and each was found to be self-incompatible on the basis of fruit set patterns. Pollen tubes in the styles of self-pollinated flowers of two of the monomorphic species, penetrated to the ovary, but no fruits resulted from selfpollinations.     

Gender specialization and factors affecting fruit set of the wind-pollinated heterodichogamous Juglans regia

Gender expression, flowering phenology, reproductive performance and factors affecting fruit set (i.e., flowering synchrony, size and distance to the nearest pollen donor) were investigated in a cultivated population of a wind-pollinated self-compatible heterodichogamous Juglans regia (Juglandaceae). Four flowering morphs, (i.e., protandrous, protogynous, male and female) were observed. The sexual functions of the protandrous and protogynous morphs were almost synchronous; however, they were not reciprocal, and the separation of male and female flowering within most monoecious individuals was not complete. Thus, within-morph mating and geitonogamous pollination may be common. The ratio of protandrous versus protogynous morphs was biased towards the protandrous morph, but the fruit set did not differ between the morphs, suggesting that the fruit set of the protandrous morph could be partly compensated by within-morph pollination. The ratio of the female flower number or fruit number to the total male catkin length was higher in the protogynous morph than in the protandrous morph and did not vary with plant size, suggesting that gender variation was not size dependent and that the sexual function of protandrous morphs was more male biased. Fruit set depended on plant size only for protandrous morphs. The fruit set of individual plants decreased with increasing distance to the nearest pollen donor regardless of morph, possibly because of pollen limitation. The fruit set of individual plants increased with flowering synchrony, indicating that flowering synchrony could affect reproductive success.     

Pollination efficiency and reproductive patterns in relation to local plant density,population size,and floral display in the rewarding Listera ovata (Orchidaceae)

Pollination efficiency and reproductive success vary strongly among populations of most animal‐pollinated plant species, depending on their size and local density, whereas individual plants within populations experience varying levels of reproductive output as a result of differences in floral display. Although most orchid species have been shown to be severely pollination limited, few studies have investigated the impact of the above‐mentioned factors on pollination success and reproduction, especially in rewarding species. In this study, the impact of population size, local density of flowering plants, and floral display on the rates of pollinia export and fruit production was investigated in 13 natural populations of the rewarding terrestrial orchid Listera ovata. In addition, an emasculation experiment was set up to examine how floral display and local density of flowering plants affected the relative importance of cross‐ vs. geitonogamous pollination in determining fruit set. In the studied populations, pollination efficiency, pollen removal, and fruit set increased with increasing population size until a threshold value of 30–40 flowering plants was reached, above which pollination efficiency and reproductive output decreased again. On average, plants with large floral displays showed higher proportional pollinia removal and fruit set compared with smaller plants. Fruit production was also significantly and positively related to local plant density, whereas emasculation did not affect the relationship between local plant density and fruit set, suggesting that geitonogamous pollination did not affect the outcome of female function. The results of this study are discussed in the light of the flowering mechanism of the species and its generalized pollination system. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 157 , 713–721.     

How pollinator-mediated mating varies with population size in plants

In most higher plants sexual interactions are mediated by animal pollinators that affect the number and differential reproductive success of mates. The number and sex of breeding individuals in populations are central factors in evolutionary theory, but the quantitative effect of plant population size on pollinator-mediated mating is understudied. We investigated variation in pollen removal (male function) and fruit set (female function) among flowering populations of different size of two bumblebee-and one butterfly-pollinated, rewardless, pollen-limited, hermaphroditic orchid species in Sweden. As the amount of pollen removed from plants by insects (either absolute or proportional) increased, so did the number of pollinations, whereas the proportions of plants with different pollinator-designated functional sex (male, female, hermaphrodite) depended primarily on the ratio between the amount of fruit set and pollen removed within populations. A larger population size was found to have several effects: (1) the total numbers of pollinia removed and fruits set increased; (2) the proportion of pollen removed from plants decreased; (3) the proportion of flowers pollinated decreased in the butterfly-but was not affected in the bumblebee-pollinated species; (4) the ratio between fruits set and pollinia removed increased linearly in the bumblebee-pollinated species but reached a maximum at c. 80 individuals in the butterfly-pollinated species; (5) the numbers of pollinator-designated pure male and hermaphrodite individuals increased; and (6) the variance in pollinium removal, but not fruit set, increased among individuals. These findings empirically verify the basic importance of population size for the mating structure of outcrossing plants, and indicate that selection for female sexual traits is reinforced when population size is smaller while selection for male sexual traits is reinforced when population size is larger.     

AN UNUSUAL KIND OF DISTYLY IN QUINCHAMALIUM CHILENSE (SANTALACEAE) ON VOLCÁN CASABLANCA,SOUTHERN CHILE

In mature flowers of the southern Andean parasitic herb, Quinchamalium chilense (Santalaceae), the stigmas and anthers are closer together in the pin morph than in the thrum morph. While the stigmas and anthers of the two morphs are in reciprocal positions as the flowers open, such reciprocity is lost as the result of post-anthesis allometric growth of the styles and stamens. Experimental pollinations reveal that both morphs of Q. chilense are self-compatible. Natural fruit set is higher on the pin morph. The latter is also statistically under-represented in natural populations with respect to a 1:1 ratio for pins and thrums. Pin flowers produce larger numbers of pollen grains than thrum flowers and pollen of pin flowers is smaller in size than that of thrum flowers. Higher fruit set on the pin morph is consistent with some tendency towards subdioecious breeding behavior, although an expected excess of geitonogamous pollinations on the pin morph might also be contributing to the difference in fruit set on the two morphs. The unusual floral morphology of Q. chilense could have evolved as a result of selection for larger flowers by certain pollinator groups less likely to effect geitonogamous pollination. Alternatively, extended development of post-anthesis flowers might be a reflection of selection for an array of flower sizes on individual inflorescences, producing a target effect for pollinator attraction. The evolution of such inflorescence morphology would have been facilitated by the fact that a trend in the direction of subdioecious breeding behavior accrues the same outcrossing advantages as strict distyly. Precise information on periods of stigma receptivity is required to further our understanding of the floral morphology of Q. chilense.