MEASURING POLLINATOR-MEDIATED SELECTION ON MORPHOMETRIC FLORAL TRAITS: BUMBLEBEES AND THE ALPINE SKY PILOT,POLEMONIUM VISCOSUM

Sweet-flowered plants of Polemonium viscosum in Colorado are visited by a fly-dominated pollinator fauna at timberline (krummholz), but almost exclusively by bumblebees in higher-elevation tundra habitats. Significant increases in flower size and height are associated with increasing elevation along this habitat gradient. This paper presents the results of an experiment designed to test whether bumblebees exert sufficient selection on morphometric floral phenotypes to account for the clinal shifts seen in natural populations. Two populations of sweet-flowered plants of krummholz origin were established: one randomly pollinated, the other solely bumblebee-pollinated. I tested the effects of two independent axes of floral variation, obtained by principal-components analysis, on mean seed set per flower of plants in each population. PC1, with strong correlations to corolla diameter, corolla length, and stem height, explained a significant amount of variance in seed set for bumblebee-pollinated plants but had no bearing on that of randomly pollinated plants. PC2, with strong correlation to flower number, did not influence seed set in either population. Bumblebee behavior was correlated with variation in PC1 scores of the selected population, yielding positive directional selection on morphometric floral traits associated with PC1. Selection coefficients for PC1, corolla length, corolla diameter, and inflorescence height were estimated, respectively, as 0.11, 0.09, 0.07, and 0.06 (P < 0.025 in all cases). These results support the hypothesis that pollinator-mediated selection can bring about changes in floral form, and can explain shifts in floral morphology of P. viscosum along natural habitat gradients.     

POLLEN DISPERSAL DYNAMICS IN AN ALPINE WILDFLOWER,POLEMONIUM VISCOSUM

The alpine wildflower, Polemonium viscosum, depends on insect visitors for effective pollination. Here, I examine experimentally the effects of pollinator visitation on pollen removal, pollen dispersal success, paternity, and gene flow. Bumble bee pollinators visited donor individuals homozygous for marker alleles at an isozyme (GOT-2) encoding locus and then were presented with arrays of recipient plants lacking the marker alleles. Four aspects of male fitness were estimated for each donor: the number of pollen grains dispersed to flowers of the first recipient visited, the number of offspring sired on that recipient, the proportion of offspring sired in the full array, and the proportion of mates in the array bearing seeds of the donor. Pollen removal was strongly influenced by the number of bee visits to donor flowers. The amount of pollen removed in turn significantly affected the number of pollen grains reaching flowers of the first recipient. However, because seed production decelerates with stigma pollen load, the relationship between pollen export and paternal success at this proximate scale showed diminishing returns. The probability of reaching mates within the array also increased with pollen export. These findings show that floral characters enhancing pollinator visitation rate in P. viscosum have positive effects on paternity and gene flow.     

RATES OF FLORAL EVOLUTION: ADAPTATION TO BUMBLEBEE POLLINATION IN AN ALPINE WILDFLOWER,POLEMONIUM VISCOSUM

Animal pollinators are thought to shape floral evolution, yet the tempo of this process has seldom been measured. I used the prediction equation of quantitative genetics, R = h²S , to predict the rate at which a change in pollinator abundance may have caused divergence in floral morphology of the alpine skypilot, Polemonium viscosum. A selection experiment determined the rate at which such divergence can actually proceed. Corolla flare in this species increases by 12% from populations pollinated by a wide assemblage of insect visitors to those pollinated only by bumblebees. To simulate the evolutionary process giving rise to this change, I used a pollinator selection experiment. Plants with broad flowers set significantly more seeds than plants with narrow flowers under bumblebee pollination but had equivalent fecundity when visited by other insects or hand-pollinated. Bumblebee-mediated selection for broad corolla flare intensified from 0.07 at seed set to 0.17 at progeny establishment. Maternal parent-offspring regression yielded a confidence interval of 0.22–1.00 for trait heritability. Given these parameter estimates, the prediction equation shows that broadly flared flowers of bumblebee-pollinated P. viscosum could have evolved from narrower ones in a single generation. This prediction is matched by an observed 9% increase in offspring corolla flare after a single bout of bumblebee-mediated selection, relative to offspring of unselected controls. Findings show that plant populations can adapt rapidly to abrupt changes in pollinator assemblages.     

POLLINATION IN FLORAL SCENT MORPHS OF POLEMONIUM VISCOSUM: A MECHANISM FOR DISRUPTIVE SELECTION ON FLOWER SIZE

Plants of Polemonium viscosum have flowers that are either sweet or skunky in scent. The two morphs are preferentially pollinated by insects of strongly contrasting body size: bumblebee queens specialize on sweet flowers, flies on skunky ones. In this study 13 characters were examined in plant specimens from five populations to identify major components of intraspecific variation in flower and inflorescence morphology and test their correlation with floral scent. Factor analysis identified four major axes of morphological variation. The first explained 22% of the variance among specimens and correlated strongly with four flower size characters: sepal length, corolla tube length, corolla lobe width, and corolla lobe length. Floral scent morphs differed significantly in the multivariate representation of flower size defined by these characters. Sweet flowers had wider corolla lobes, longer corolla tubes, and longer sepals than skunky ones. Corolla lobe width accounted for the greatest amount of intermorph divergence. Divergence in flower size between morphs was maintained in mixed populations at four locations in alpine Colorado, with corollas of sweet flowers significantly broader or more flared than those of skunky flowers. Patterns of pollen receipt suggest that this difference is adaptive. In the sweet morph, pollination intensity and purity increased significantly with corolla flare. Conversely, in the skunky morph, corolla flare had little influence on pollination intensity and had a strong negative effect on purity. These findings suggest that selection for effective pollination should favor intraspecific divergence in flower size in Polemonium viscosum.     

VARIATION IN SEX ALLOCATION AND FLORAL MORPHOLOGY IN IPOMOPSIS AGGREGATA (POLEMONIACEAE)

Intrapopulational variation in biomass allocation to male vs. female function was quantified for the hermaphroditic plant Ipomopsis aggregata in terms applicable to sex allocation models. The proportions of flower biomass put into the corolla and calyx averaged 0.59 and 0.20 and were relatively constant across plants. The proportions in the stamens and pistil averaged 0.13 and 0.08, with considerable variation among plants. Phenotypic gender at the time of flowering ranged from 0.34 to 0.77 female. Pistil dry weight was correlated with stigma exsertion. Stamen weight was correlated with corolla width, which influences male pollination success, and was also correlated with anther position and pollen production. Female reproductive success as estimated by seeds per flower showed no detectable relationship with initial allocation of biomass at the time of flowering, but decreased in accelerating fashion with the proportion of final biomass including seeds that was allocated to male function.     

DOWN THE TUBE: POLLINATORS, PREDATORS, AND THE EVOLUTION OF FLOWER SHAPE IN THE ALPINE SKYPILOT, POLEMONIUM VISCOSUM

Abstract We address how a conflict between pollinator attraction and avoidance of flower predation influences the evolution of flower shape in Polemonium viscosum. Flower shape in P. viscosum is the product of an isometric relationship between genetically correlated (r_A= 0.70) corolla flare and length. Bumblebee pollinators preferentially visit flowers that are more flared and have longer tubes, selecting for a funnel‐shaped corolla. However, flower shape also influences nectar‐foraging ants that sever the style at its point of attachment to the ovary. Surveys of ant damage show that plants having flowers with flared, short corollas are most vulnerable to ant predation. Consistent with this result, the ratio of corolla length to flare is significantly greater in a krummholz (high predation risk) population than in a tundra (low predation risk) population. To explicitly test whether the evolution of a better defended flower would exact a cost in pollination, we created tubular flowers by constricting the corolla during development. Performance of tubular flowers and natural controls was compared for defensive and attractive functions. In choice trials, ants entered control flowers significantly more often than tubular ones, confirming that the evolution of tubular flowers would reduce the risk of predation. However, in a bumblebee‐pollinated population, tubular flowers received significantly less pollen and set fewer seeds than controls. A fitness model incorporating these data predicts that in the absence of the genetic correlation between corolla length and flare, intermittent selection for defense could allow tubular flowers to spread in the krummholz population. However, in the tundra, where bumblebees account for nearly all pollination, the model predicts that tubular flowers should always confer a fitness disadvantage.     

ECOTYPIC DIVERGENCE IN ALPINE POLEMONIUM VISCOSUM: GENETIC STRUCTURE,QUANTITATIVE VARIATION,AND LOCAL ADAPTATION

Polemonium viscosum has a continuous distribution from 3,500 m in the krummholz to 4,025 m on the summit ridges of Pennsylvania Mountain, Colorado. Seeds produced by plants at opposite ends of this cline, 1.5 km apart, differed significantly at allozyme loci in two consecutive breeding seasons. Mean multilocus F_st values for both years (0.015 and 0.069) were significantly different from zero, indicating restricted gene exchange between subpopulations. Average allele frequencies at two individual loci also differed significantly between families comprising krummholz and summit subpopulations. Progeny of plants growing on the summit had higher leaf production rates, more densely packed leaflets, and lower resistance to aphids than progeny of plants growing in the krummholz site, when tested under greenhouse conditions. These differences probably reflect the restricted opportunities for growth and severe exposure at high elevations, and the increased risk from herbivores near timberline. The two subpopulations did not differ in leaf length (stature), leaf width, or pubescence. Reciprocal transplanting of seedlings between krummholz and summit sites confirmed that the differences were adaptive, since progeny from each subpopulation performed significantly better in their parent's habitat. Coordinated studies of genetic structure, quantitative variation, and local adaptation across the elevational range of P. viscosum provide a comprehensive view of ecotypic divergence in this widespread alpine plant.     

POLLEN AND GENE DISPERSAL: THE INFLUENCES OF COMPETITION FOR POLLINATION

Pollinators that forage indiscriminately can transfer pollen from one species to another, reducing the amount that reaches conspecific flowers. I present evidence that the presence of another plant species visited by the same pollinators can also reduce pollen dispersal distances and outcrossing. This has the potential to influence gene flow and reproductive success. Pollen carryover and movement patterns were measured for the shared insect pollinators of Stellaria pubera and Claytonia virginica in North Carolina. Bee flies deposited similar amounts of Stellaria pollen on a series of pistillate Claytonia flowers as on a series of pistillate Stellaria flowers. In arrays of potted plants, flies and solitary bees visited most flowers on a plant before leaving and then flew to a nearby plant chosen independently of species; 95% of moves were to one of 12 nearest neighbors. These measures of pollen carryover and movement patterns were used in a set of computer simulations to predict pollen dispersal distances. The simulations suggested that C. virginica substantially reduces outcrossing and pollen flow in S. pubera. These predictions were tested by tracking dye movement from anthers in populations of potted plants. Addition of C. virginica reduced the mean squared distance moved by dye to receptive S. pubera flowers by 23% and reduced the amount of dye moved by 51%. The estimated pollen component of gene flow was also much lower in a natural population of 5. pubera mixed with C. virginica than in the synthetic single-species populations.     

SEED-COAT MORPHOLOGY AND ANATOMY IN COLLOMIA (POLEMONIACEAE)

Seed-coat morphology of 14 species of Collomia (Polemoniaceae) was examined with light microscopy and scanning electron microscopy. Two seed types were observed based on surface sulpturing: Type 1—hexagonal epidermal cells forming a shallow reticulum with well-defined cell boundaries; Type 2—longitudinally ridged and irregularly arranged crater-like depressions with inconspicuous cell boundaries. Only two species, C. debilis and C. larsenii of sect. Collomiastrum have seed-coat Type 1. Seed-coat Type 2 is characteristic of all species of sect. Collomia, sect. Courtoisia, and C. mazama, C. rawsoniana of sect. Collomiastrum. The present investigation reveals a fairly homogeneous seed-coat pattern in the genus and does not offer significant information for realignment of infrageneric classification. Anatomical studies with light microscopy show that a mucilaginous seed coat develops from the outermost layer of integument in which each epidermal cell develops spiral secondary wall thickenings. Mucilaginous seeds of most Collomia species probably provide adaptive significance in that adherence of seeds to ground prohibits further dispersal to unfavorable habitats, or epizoochoric dispersal.     

POLLEN MORPHOLOGY AND ULTRASTRUCTURE OF THE CABOMBACEAE: CORRELATIONS WITH POLLINATION BIOLOGY

Of all species comprising the two genera of the Cabombaceae, only Brasenia schreberi J. F. Gmel. and Cabomba caroliniana Gray have been critically investigated with regard to their pollination biology. Brasenia schreberi has been shown to be anemophilous, while C. caroliniana has an entomophilous (myophilous) pollination syndrome. In the present paper, a number of pollen and pollen-related characters, including pollen size, shape, quantity, terminal settling velocity, pollen-ovule ratios, and overall exine architecture of B. schreberi and C. caroliniana are evaluated. Pollen from both species is elliptic, monosulcate, and has a tectate-columellate sporoderm with supratectal surface ornamentation. Grains of B. schreberi are small, produced in copious amounts, and settle relatively slowly. Flowers of this species have large pollen-ovule ratios. The exine of B. schreberi pollen is scabrate, relatively thin, has a uniformly thick sexine composed of a two-zoned (homogeneous/granular) tectum and distinct columellae, and a homogeneous nexine. Pollen of C. caroliniana is relatively large, produced in small quantities, and has a rapid terminal settling velocity. Flowers exhibit small pollen-ovule ratios. Exine organization of C. caroliniana pollen is typically two times thicker than that of B. schreberi; ornamentation is striate. Nonapertural sexine regions have a thick tectum and well-defined columellae, with both sexine components traversed by a dense system of channels. The nexine is relatively thin. All of the palynological characters examined correlate well with the anemophilous and entomophilous syndromes of B. schreberi and C. caroliniana, respectively. Moreover, several other parameters of exine ultrastructure from each species exhibit positive correlations with the respective pollination mechanisms, including: tectum thickness, columellae diameter, tectum-nexine ratios, and the consistency, distribution, and total amount of pollenkitt present. Overall exine ultrastructure is also discussed from a historical perspective as well as with respect to its phylogenetic significance.     

POLLINATION AND THE FUNCTION OF FLORAL PARTS IN CHAMAECRISTA FASCICULATA (FABACEAE)

Chamaecrista fasciculata has a rigid upper petal that curves over nine of the ten anthers. The single anther is deflexed, as is the style. An experimental pollination study was designed to test the hypotheses that: 1) the rigid petal acts as flight guide to ensure pollination; and 2) the nine grouped anthers serve as fodder anthers, whereas the single deflexed anther functions in pollination. The rigid petal was removed from 97 flowers. Only 5% of the manipulated flowers set fruit in comparison to a fruit set of 47% for the control group. The results of the study support the flight guide hypothesis. Pollen from both sets of anthers is viable and germinates on receptive stigmas. A pollen flow experiment using powdered metals, backscatter scanning electron microscopy, and X-ray microanalysis revealed that there is no difference in the frequency of pollen distribution from either set of anthers. Therefore, this study refutes the fodder/pollinating anther hypothesis.     

POLLEN MORPHOLOGY IN THE GENERA MONTTEA AND MELOSPERMA (SCROPHULARIACEAE)

Pollen of the scrophulariaceous genera Melosperma and Monttea was studied with light and scanning electron microscopy. Both have single, radiosymmetric, isopolar, trizoniaperturate pollen grains with usually ruptured colpus membranes, tectate-perforate to semitectate exines with a columellate interstitium and pitted or perforate nexines. Intergeneric differences in pollen morphology are established based on the differential correlation of sculpturing characters. Comparisons of the pollen of Melosperma and Monttea with that of the tribe Minuleae sensu stricto reveals no difference in the case of Melosperma and the existence of intermediate morphologies and “reticulate” similarities in the case of Monttea; the taxonomic significance of this morphological intergradation is considered. The transfer of Oxycladus aphyllus to Monttea is consistent with the pollen evidence, but proposed affinities between Monttea and tribe Antonieae of the Loganiaceae are not supported.     

江淮丘陵区早春开花植物的花粉形态与其生态因子

应用光学显微镜对安徽省枞阳县和安徽大学校园内春季开花的22科30属40种植物的花粉形态进行了观察和研究.结果表明花粉粒扁球形至长球形,P/E值最大的花粉见于贴梗海棠[Chaenometes speciosa (Sweet)Na-kai],为1.41(1.23-1.79),P/E值最小的花粉见于枫杨(Pterocarya stenoptera C. 13(2.),为0.78(0.71-0.83);体积最大的花粉见于金银花(Lonicera japonica Thunb),直径为60.8(57.5-65.0)μm,体积最小的花粉见于菱叶绣线菊(Arisaema jacquemontii Blume),为11.5(10.0-12.5)×9.8(8.0-12.)μm.萌发孔主要有3沟(20.5%)、3孔沟(61.5%)、散孔(15.4%),六孔(2.6%)等类型.外壁纹饰主要有条纹-穿孔(22.5%)、细网状(27.5%)、粗网状(25.0%)、条纹状(50%)和颗粒状(12.5%)等类型.同时调查了其生态因子,包括地理位置、海拔高度、年降水量、年积温及生境等,得出了这些植物分布区的主要生态因子.该花粉组合信息可用以重建古植被、古气候及古环境的变化.     

安徽鹞落坪国家自然保护区植物的花粉形态与其生态因子

应用光学显微镜对安徽省鹞落坪国家级自然保护区7月份开花的25科41属47种1变种植物的花粉形态进行了观察和研究.结果表明花粉粒扁球形至超长球形,P/E值最大的为安徽贝母(Fritillaria anhuiensis S. C. Chen et S. F. Yin),大小为1.71(1.28-2.30)μm,最小的为粉花绣线菊(Spiraea japonica L.),大小为0.83(0.58-0.97)μm;花粉最大的为萱草[Hemerocallis fulva (L.)L.],大小为93.8(85.0-100.0)×61.1(51.3-70.0)μm;最小的为草绣球[Cardiandra moellendor f fii (Hance) Migo],大小为11.1(9.5-12.5)×8.8(7.5-10.0)μm.萌发孔主要有单沟(8.3%)、3-4沟(18.8%)、多沟(8.3%)、3-4孔沟(58.3%)、散孔(4.2%)、无萌发孔(2.1%)等类型.外壁纹饰主要有刺状(12.5%)、细网状(33.3%)、粗网状(16.7%)、颗粒状(25.0%)及无明显纹饰(12.5%)等类型.同时研究了其生态因子,包括地理位置、海拔高度、年降水量、年积温及生境等,特别提供了7月份的气候因子,得出这些花粉分布区的主要生态因子.为利用地层中相应化石花粉重建大别山地区古植被、古环境及气候变迁提供了现代孢粉学资料和依据,也为这些植物的现代地理分布提供了科学依据.     

安徽省皖南山区植物的花粉形态和生态因子

应用光学显微镜和扫描电镜研究安徽省皖南山区广德柏垫和东至利安5月份开花的37科66种植物的花粉形态.对这些植物的生态因子,包括海拔高度、年降水量、年积温和生境进行归纳.进而根据同一区域内相同或相似生态环境条件下分布的植物花粉类型,寻取一定花粉类型综合特征与气候和环境之间可能的联系,为地层孢粉研究提供现代孢粉学依据.