DIFFERENTIAL SUCCESS OF POLLEN DONORS IN A SELF-COMPATIBLE LILY

If pollen donors are equally effective at siring seeds, the presence of equal proportions of pollen from two pollen donors on a stigma will lead to equal proportions of seeds sired by each pollen donor. Variation in germination rates, pollen-tube growth, and embryo viability may cause one donor to sire more seed than another. We looked for differential donor success in the field by simultaneously applying equal amounts of pollen from two pollen donors. We simultaneously applied equal amounts of self and outcross pollen to receptive stigmas and simultaneously applied pollen from two donors at different physical distances from the recipient. Following simultaneous application of self and outcross pollen, significantly more of the seeds were sired by outcross pollen donors. Seed set following simultaneous application of two outcross donors was also nonrandom. Pollen donors from 100 m were more likely to sire seeds when competing with pollen from plants nearby (1 m). To determine whether pollen-tube growth rates were responsible for these patterns of paternity, we varied the timing of deposition of outcross pollen allowing self pollen tubes a head start on the stigma. Outcross pollen was applied 3 or 24 h after self pollen. In spite of this time delay, the majority of the seeds were again sired by outcross pollen. There was no significant difference in the amount of seeds sired by self pollen between the two delay treatments. This result suggests that mechanisms operating after ovule fertilization may contribute to the discordance between the proportions of the pollen present and the proportions of seeds sired.     

Effects of pre-dispersal selection on offspring growth and survival in Erythronium grandiflorum

Effects of pre-seed-dispersal processes on offspring vigor were examined in Erythronium grandiflorum using manipulations of the number of pollen donors contributing to the pollen pool and comparisons of means and variances in offspring growth measurements. There were no effects of the number of donors on measures of pollen-tube growth, ovule abortion, seed set, mean seed weight, or seedling germination. Seeds from pollinations with only one donor produced corms that averaged 5% lighter after one season of growth and had lower overall survival after three years compared to corms from pollinations with either three or ten donors. Patterns of within- and among-family variance estimates for the different treatments were consistent with the hypothesis that less-vigorous offspring were eliminated prior to seed dispersal in the multiple-donor treatments. The difference in the growth of offspring from different treatments was apparently not due to pollen competition because pre-zygotic attrition of pollen tubes led to incomplete fertilization of ovules. Results from this study suggest that post-fertilization abortion of less-vigorous progeny, perhaps as a consequence of early-acting inbreeding depression, is responsible for the increase in the average vigor of offspring from multiply-sired fruits.     

Effects of pollen load size on seed paternity in wild radish: the roles of pollen competition and mate choice

For sexual selection to be important in plants, it must occur at pollen load sizes typical of field populations. However, studies of the impact of pollen load size on pollen competition have given mixed results, perhaps because so few of these studies directly examined the outcome of mating when pollen load size was varied. We asked whether seed paternity after mixed pollination of wild radish was affected by pollen load sizes ranging from 22 to 220 pollen grains per stigma. We examined the seed siring abilities of 12 pollen donors across 11 maternal plants. Seed paternity was statistically indistinguishable across the pollen load sizes even though, overall, the pollen donors sired different numbers of seeds. This lack of effect of pollen load size on seed paternity may have occurred because fruit abortion and early abortion or failure of fertilization of seeds increased as load size decreased. Thus, failures of fruits and seeds sired by poorer pollen donors may keep seed paternity constant across pollen load sizes.     

The timing and effectiveness of sequential pollinations in Hibiscus moscheutos

In many species with animal-pollinated flowers, pollen arrives on the stigma in pulses and late-arriving pollen may be precluded from fertilizing ovules. When seed set per fruit is not pollen limited, the fate of pollen from later cohorts is affected by the time between pollinator visits, variation in pollen tube growth rates, and the amounts of pollen deposited relative to the availability of stigmatic area and unfertilized ovules. In a natural population of Hibiscus moscheutos, we found that consecutive pollinator visits to individual flowers occurred within 15 min of each other in more than half of our observations. We then conducted hand-pollination experiments using equal doses of early and late pollen bearing unique allozyme markers for paternity analysis (each dose was more than sufficient to result in full seed set). When pollen was applied 15 or 30 min after an earlier pollen load, the proportions of seeds sired by late-arriving pollen were reduced by 13–30% and 21–57%, respecitively. A few pollen-tubes from the late pollen load sired seeds even when a large dose of pollen was applied 1–2 h earlier, suggesting that the performance of pollen grains within each cohort was highly variable. The advantage of arriving early was greater when the first pollen load was applied at 0930 hours as compared with 1130 hours. We found no effect of previous pollination on the growth rates of late-arriving pollen-tubes. Taken together, these results demonstrate that pollen from later visits can compete with pollen from earlier visits, but the success of late-arriving pollen typically declines to very low levels after an interval of about 2 h. Given that surplus pollen often arrives on stigmas by mid-morning, we conclude that male reproductive success is more likely to be affected by the timing of pollen dispersal and pollen-tube competitive ability than by the total amount of pollen that is exported from flowers.     

Effects of temperature and pollination site on pollen performance in Betula pendula Roth – evidence for genotype-environment interactions

We studied whether the differences between genetically different pollen donors (Betula pendula Roth clones) with respect to pollen-tube growth rate were consistent under different thermal conditions during pollen germination in vivo and in vitro. We conducted a single-donor hand-pollination experiment with same pollen donors and recipients in a plastic house seed orchard and at an outdoor clone collection. The prevailing daily mean temperature during pollen germination was 13°C higher in the plastic house than outdoors. The pollen-tube growth rate of each pollen donor was additionally determined in vitro on agar medium at five temperatures (10°, 15°, 22°, 30° and 35°C). A significant interaction between paternal clone and pollination site as well as between paternal clone and temperature was found, which provides evidence for genotype-environment interactions. Genotype-environment interactions can have evolutionary significance in maintaining the variation in pollen-tube growth rates. At seed orchards, genotype-environment interactions can cause deviations from the expected genetic composition of the seed crop depending on the prevailing environmental conditions during pollen-tube growth. Received: 24 September 1999 / Accepted: 29 September 1999     

Pollen-tube growth rate and seed-siring success among Betula pendula clones

The aim of this study was to investigate whether genetically different pollen donors ( Betula pendula clones) differed in pollen-tube growth rate across 11 maternal plants and in vitro , and whether the differences between the donors were consistent across the recipients. To compare the seed-siring success of competing pollen donors, a two-donor hand-pollination experiment with six donors and six recipients was conducted. The experiments were performed at a plastic-house seed orchard. The donors showed significant variation in pollen-tube growth rate on all the 11 recipients. The rankings of the pollen donors were statistically consistent across different maternal plants. A significant positive correlation between pollen tube growth in vivo and in vitro was found. The seed-siring success of two competing pollen donors was unequal in 20 of 29 cases and there was a significant positive correlation between seed-siring success and pollen-tube growth rate in vivo and in vitro . The results show that fertilizations are not random and pollen competition operates in a B. pendula seed orchard population.     

Effects of pollination timing on seed paternity and seed mass in Silene latifolia (Caryophyllaceae)

Background and Aims

Competition among genetically different pollen donors within one recipient flower may play an important role in plant populations, increasing offspring genetic diversity and vigour. However, under field conditions stochastic pollen arrival times may result in disproportionate fertilization success of the first-arriving pollen, even to the detriment of the recipient plant''s and offspring fitness. It is therefore critical to evaluate the relative importance of arrival times of pollen from different donors in determining siring success.

Methods

Hand pollinations and genetic markers were used to investigate experimentally the effect of pollination timing on seed paternity, seed mass and stigmatic wilting in the the dioecious plant Silene latifolia. In this species, high prevalence of multiply-sired fruits in natural populations suggests that competition among different donors may often take place (at fertilization or during seed development); however, the role of variation due to pollen arrival times is not known.

Key Results

First-arriving pollen sired significantly more seeds than later-arriving pollen. This advantage was expressed already before the first pollen tubes could reach the ovary. Simultaneously with pollen tube growth, the stigmatic papillae wilted visibly. Individual seeds were heavier in fruits where one donor sired most seeds than in fruits where both donors had more even paternity shares.

Conclusions

In field populations of S. latifolia, fruits are often multiply-sired. Because later-arriving pollen had decreased chances of fertilizing the ovules, this implies that open-pollinated flowers often benefit from pollen carry-over or pollinator visits within short time intervals, which may contribute to increase offspring genetic diversity and fitness.Key words: Reproduction, reproductive success, pollen, siring success, microsatellite DNA, paternity, pollen tube growth, seed mass, Silene alba, stigma wilting     

Pollen load size, reproductive success, and progeny kinship of naturally pollinated flowers of the tropical dry forest tree Pachira quinata (Bombacaceae)

Several studies have demonstrated, using controlled pollinations, that the number and identity of pollen grains deposited onto a flower's stigma affect the reproductive success of plants. However, few studies have shown this relationship under conditions of natural pollination. Using the tropical dry forest tree Pachira quinata, we evaluated the relationship between the number of microgametophytes per pistil and the number of sires with respect to the production of fruits and seeds in a natural population of Pachira quinata. Our study demonstrates that fruit and seed production are directly related to the number of microgametophytes per pistil in natural populations of P. quinata. Only 6% of the marked flowers developed into mature fruits and 10% of the marked flowers initiated fruits but later aborted them. A mean of 23 pollen grains were required to produce a seed. Flowers with >400 pollen grains on the stigma always developed into mature fruits, whereas flowers that received <200 grains never matured fruits. Half of the pollen grains transferred to a flower stigma germinated and developed pollen tubes to the base of the style. The number of pollen grains on a stigma explained 34% of the variation in seed number per fruit, and the number of seeds produced per fruit is positively correlated with the size of the seeds. The population of P. quinata studied is predominantly outcrossing, and seeds within fruits are sired by one or a few donors. The total seed crop within trees was sired by three to five donors. Our study examined the implications of the variation in size of microgametophyte loads per pistil with respect to the breeding system and the paternity of progeny under natural conditions. The competitive ability of pollen and pollen tube attrition are important factors regulating fruit production in P. quinata.     

VARIATION IN POLLEN SIZE,FERTILIZATION ABILITY,AND POSTFERTILIZATION SIRING ABILITY IN ERYTHRONIUM GRANDIFLORUM

The mean volume of pollen grains and total pollen production varied both within and among plants of Erythronium grandiflorum. The second flowers of two-flowered plants tended to produce smaller and fewer grains than first flowers, but there was no overall relationship between mean pollen grain size and production per flower. I evaluated the effects of pollen size differences within and among plants on two components of male reproductive success: pollen tube growth and postfertilization siring ability. Pollen tubes grown in media were longer for second flowers, but were not correlated with the mean size of pollen grains, suggesting that (1) internal resource content of pollen (i.e., carbohydrates plus lipids) was not associated with the hydrated size of pollen, and that (2) pollen from second flowers contained more resources. I analyzed the growth rate and the fertilization ability of pollen growing in styles. Growth rate differed among donors and recipients, but no effects of pollen or donor characters (i.e., pollen production, grain size, and flower position) were detected. In single donor pollinations, pollen size was negatively correlated with fertilization ability across donors, and positively correlated with postfertilization siring ability of donors. A second experiment used pairs of donors; within-plant differences in pollen size and flower position had effects similar to the single donor experiment on fertilization ability, but among-plant differences were not significant. The results corroborate earlier experiments that suggest that the growth of pollen tubes in the style is probably controlled by the recipient, since donor characters had minimal effects on pollen fertilization ability. Postfertilization siring ability was not affected by within-plant differences in mean grain size and production. For among-donor differences, the number of seeds set for each donor was positively correlated with the mean grain volume, and when a donor producing large pollen fertilized ovules in an ovary, there was increased seed abortion for seeds in the same ovary sired by a second donor. In addition, the total number of seeds produced by a fruit was decreased when both donors had large pollen, apparently due to increased postfertilization abortion. Postfertilization processes appear to be influenced by paternal differences that are expressed through competition among developing seeds for maternal resources.     

POLLEN-TUBE COMPETITION AND MALE FITNESS IN HIBISCUS MOSCHEUTOS

The stigmas of animal-pollinated flowers often capture more pollen than is needed to fertilize all available ovules, and mixed-donor pollen loads are probably common. When this is the case, variation in average pollen-tube growth rates can potentially affect the number of seeds sired by a given plant. Despite considerable interest in effects of postpollination processes on male fitness, little is known about the extent of variation in pollen performance among plants from natural populations. To examine this question in Hibiscus moscheutos (rose mallow), we conducted mixed-donor hand-pollination experiments with 39 pollen donors bearing distinctive isozyme markers. Pairs of competing donors were compared on sets of 11 to 15 recipient plants per pair. These donors often differed in the proportions of seeds they sired, with the maximum deviation from an expected ratio of 50:50 being 68:32. Furthermore, three intensively studied plants exhibited consistent trends in relative pollen performance when each was tested against (1) the same three competitors, and (2) groups of 14 competitors chosen at random from the study population. In a separate experiment, we investigated the effects of salinity stress and high soil nutrients on pollen performance. These environmental factors had anticipated effects on leaf production, flower production, and petal length, but style length and (most importantly) the number of seeds sired relative to a standard pollen donor were not affected. In summary, this study provides the strongest evidence to date that pollen-tube competitive ability varies among coexisting plants and may be an important component of male fitness in plants.     

Under how wide a set of conditions will nonrandom mating occur in Raphanus sativus (Brassicaceae)?

Studies of the weedy annual Raphanus sativus have demonstrated that nonrandom mating, a prerequisite for sexual selection, can occur in greenhouse plants. To determine whether this nonrandom mating pattern can occur under a wide range of conditions, including conditions that might occur in the field, we considered variation in both maternal condition and pollen load size. Maternal condition was varied by altering the watering regime. Pollen load size was varied from approximately 26 to 343 pollen grains per stigma. At the smallest pollen load size, patterns of seed paternity were altered in two of the three pollen donor pairs; seed paternity became more equal among donors. For one of three pollen donor pairs, seed paternity was more divergent among donors on stressed maternal plants. Finally, for one pollen donor pair, rank order of pollen donor performance changed from the medium to the small pollen loads on stressed vs. control maternal plants. Thus, some field conditions may alter patterns of nonrandom mating in wild radish.     

Herkogamy and mate diversity in the wild daffodil Narcissus longispathus: beyond the selfing–outcrossing paradigm in the evolution of mixed mating

Spatial separation of male and female reproductive structures (herkogamy) is a widespread floral trait that has traditionally been viewed as an adaptation that reduces the likelihood of self‐pollination. Here we propose that increased herkogamy may also influence another important aspect of plant mating: the diversity of pollen donors siring seeds within fruits. We test this hypothesis in Narcissus longispathus, a wild daffodil species with extensive variation in anther–stigma separation. To study the morphological basis of variation in herkogamy, floral measurements were undertaken in 16 populations of N. longispathus. We then quantified multilocus outcrossing rates and the correlation of outcrossed paternity in three of these populations sampled over several years. Mating system estimates were calculated for each population and year, and also separately for groups of plants that differed markedly in herkogamy within each population and year. In N. longispathus herkogamy was much more variable than other floral traits, and was more closely related to style length than to anther position. Averaged across populations and years, plants with high herkogamy had similar outcrossing rates (0.683) to plants with intermediate (0.648) or low herkogamy (0.590). However, a significant linear trend was found for correlation of outcrossed paternity, which increased monotonically from high herkogamy (0.221), through intermediate herkogamy (0.303) to low herkogamy (0.463) plants. The diversity of pollen donors siring seeds of high herkogamy Narcissus flowers was thus consistently greater than the diversity of pollen donors siring seeds of low herkogamy flowers. Results of this study contribute to the emerging consensus that floral traits can simultaneously influence several aspects of plant mating system in complex ways, thus extending the traditional focus centred exclusively on patterns and relative importance of self‐ and cross‐fertilisation.     

Pollen competition and seed-siring success in Picea abies

The aim of the present work was to study pollen-tube competition in Picea abies. Controlled crossings were performed with pollen mixtures including pairs of pollen lots with fast and slowly elongating pollen-tubes. Paternity analysis using isozyme markers was performed on the progenies in order to study whether the in vitro pollen-germination vigour corresponds to the proportion of seeds sired by the pollen donor. Paternal success was found to be unequal, 15 out of 23 crossings producing progeny that differed significantly from the hypothetical ratio of 1:1. The paternal contribution in the majority of the crossings was as expected: the pollen parent with more-vigorous in vitro germination sired more seeds than the less-vigorous pollen. In the case of two pollen mixtures, however, the seed-siring success summed over the maternal trees was the opposite to the expected value. Despite these aberrations, the results support the hypothesis that pollen-tube competition is one of the factors contributing to male fitness in P. abies. However, when all the other factors affecting pollination and seed set under natural conditions are taken into account, it is clear that the seed-siring success of a particular paternal genotype cannot be predicted reliably by measuring only the in vitro pollen vigour. Received: 2 July 2001 / Accepted: 7 August 2001     

Relationships of pollen size,pistil length and pollen tube growth rates in Rhododendron and their influence on hybridization

Summary Pollen size and pistil length data have been collected for 93 species of Rhododendron (Ericaceae) belonging to a number of different subgeneric taxa. For a sample of eight species in section Vireya, pollen tube growth in the style after selfor interspecific pollination has been quantified. Pollen volume and the time taken for pollen tubes to reach the ovary were both related to pistil length. Pollen-tube growth rates were generally greater for species with longer pistils and larger pollen. Increasing temperature increased the rate of pollen-tube growth. There was no detectable effect of pollen tube density on tube growth rate in the style. After interspecific pollinations tube growth rates in foreign styles could be faster or slower than in self styles. A semisterile individual with two viable pollen grains per tetrad and a plant grafted as scion to a longer-styled stock both showed more rapid pollen-tube growth than expected on the basis of pistil size. Data collected for 26 species in section Vireya showed that where extreme disparity of pollen/pistil size causes failure of interspecific crosses, one or more bridging species with intermediate pollen/pistil size can generally be selected.     

Does multiple paternity affect seed mass in angiosperms? An experimental test in Dalechampia scandens

Flowers fertilized by multiple fathers may be expected to produce heavier seeds than those fertilized by a single father. However, the adaptive mechanisms leading to such differences remain unclear, and the evidence inconsistent. Here, we first review the different hypotheses predicting an increase in seed mass when multiple paternity occurs. We show that distinguishing between these hypotheses requires information about average seed mass, but also about within‐fruit variance in seed mass, bias in siring success among pollen donors, and whether siring success and seed mass are correlated. We then report the results of an experiment on Dalechampia scandens (Euphorbiaceae), assessing these critical variables in conjunction with a comparison of seed mass resulting from crosses with single vs. multiple pollen donors. Siring success differed among males when competing for fertilization, but average seed mass was not affected by the number of fathers. Furthermore, paternal identity explained only 3.8% of the variance in seed mass, and siring success was not correlated with the mass of the seeds produced. Finally, within‐infructescence variance in seed mass was not affected by the number of fathers. These results suggest that neither differential allocation nor sibling rivalry has any effect on the average mass of seeds in multiply sired fruits in D. scandens. Overall, the limited paternal effects observed in most studies and the possibility of diversification bet hedging among flowers (but not within flowers), suggest that multiple paternity within fruits or infructescence is unlikely to affect seed mass in a large number of angiosperm species.     

Analysis of pollen-style interactions in Petunia hybrida; the determination of variance in male reproductive success

Summary A factorial cross design was used to evaluate the influence of differences among pollen donors and recipients on variation in stylar attrition of pollen tubes in self-fertile plants of Petunia hybrida. Pollinations to flower buds were used to reduce the inhibitory ability of the style and these crosses were compared with flower pollinations to assess the degree of stylar influence on pollen fertilization ability. There was less pollen tube attrition after bud pollinations than after flower pollinations, indicating that styles of buds were less able to inhibit pollen tubes. The variance component for plants acting as pollen donors tended to be greater after flower pollinations than after bud pollinations. The lower variance in male success after bud pollinations indicates that differences among pollen donors after flower pollinations were due to stylar inhibition of pollen rather than differences in pollen vigor. Since the level of variation in pollen growth after pollination to flowers was greater among clones than among ramets within clones, the differences in pollen fertilization ability are probably genetically based.     

EFFECTS OF DIFFERENTIAL POLLEN-TUBE GROWTH ON HYBRIDIZATION IN THE LOUISIANA IRISES

To elucidate the importance of hybridization in evolution, it is necessary to understand the processes that affect hybridization frequency in nature. Here we focus on postpollination, prefertilization isolating mechanisms using two hybridizing species of Louisiana iris as a study system. We compared the effects of differential pollen-tube growth on the frequency of F₁ hybrid formation in experimental crosses between Iris fulva and Iris hexagona. Analyses of seed production in fruits from pure conspecific and heterospecific pollinations revealed that more seeds were produced in the top half than the bottom half of fruits for all four crosses. Heterospecific pollen was applied to flowers of each species at zero to 24 h prior to conspecific pollen, thereby giving a head start to the foreign pollen. Using diagnostic isozyme markers, the frequency of hybrid progeny was examined at the level of the whole fruit and separately for the top and bottom halves of fruits. In both species, the proportion of hybrid seeds per fruit increased significantly with increasing head starts, suggesting that differences in pollen-tube growth rates affect the frequency of hybridization. In I. fulva fruits, the increase in hybrid seeds occurred in both halves of the fruits, but in I. hexagona an increase was only detected in the top half of fruits. These findings are consistent with a model that assumes attrition of pollen tubes due to the greater length of I. hexagona styles. While pollen-tube growth rate appears to be the most important factor affecting hybridization frequency in I. fulva, both pollen-tube growth rate and pollen-tube attrition appear to be important in I. hexagona.     

Direct and indirect responses to selection on pollen size in Brassica rapa L.

Pollen size varies little within angiosperm species, but differs extensively between species, suggesting the action of strong selection. Nevertheless, the potential for genetic responses of pollen size to selection, as determined by additive genetic variance and genetic correlations with other floral traits, has received little attention. To assess this potential, we subjected Brassica rapa to artificial selection for large and small pollen during three generations. This selection caused significant divergence in pollen diameter, with additive genetic effects accounting for over 30% of the observed phenotypic variation in pollen size. Such heritable genetic variation suggests that natural selection could effect evolutionary change in this trait. Selection on pollen size also elicited correlated responses in pollen number (–), flower size (+), style length (+), and ovule number (+), suggesting that pollen size cannot evolve independently. The correlated responses of pollen number, flower size and ovule number probably reflect the genetically determined and physically constrained pattern of resource allocation in B. rapa. In contrast, the positive correlation between pollen size and style length may represent a widespread gametic‐phase disequilibrium in angiosperms that arises from nonrandom fertilization success of large pollen in pistils with long styles.     

Pollen tube behavior in yellow passion fruit following compatible and incompatible crosses

Pistil squashes were used to examine the growth of Passiflora edulis f. flavicarpa Deg pollen tubes in self-compatible and self-incompatible crosses. Compatible tubes typically showed a uniform layer of callose deposition in the walls and in small plugs spaced at regular intervals within the tube. Two sites of rejection were observed in incompatible crosses: the stigma and on some occasions the style, especially in anomalous crosses. In the style the inhibition of pollen-tube growth occurred in both the upper and middle parts of the transmitting tract. These findings are consistent with the hypothesis that suggests the presence of a gametophytic gene which acts in association with the sporophytic S-gene in P. edulis. Received: 12 October 1998 / Accepted: 2 December 1999     

Nonrandom mating inDiscaria americana (Rhamnaceae)

Nonrandom mating takes place when seed paternity differs from that produced by random success of available pollen. To test if some form of this phenomenon operates in the self-incompatible perennialDiscaria americana, we performed one-donor hand pollinations involving seven individuals, and measured pollen-tube performance and seed viability and germination. A significant level of heterogeneity in reproductive traits was present in our sample. Mating was nonrandom as a consequence of complete self-incompatibility (intrafloral and geitonogamous pollinations were unsuccessful) linked with partial cross-compatibility (some inter-individual pollinations consistently failed to produce offspring while other parental combinations were fecund). Pollen tubes of successful pollen parents exhibited higher growth rate than those of infecund ones. Stylar-ovarian inhibition of incompatible pollen tubes and a wet stigma type suggests that a gametophytic incompatibility system operates in the species.