The effect of petal-size manipulation on pollen removal,seed set,and insect-visitor behavior in Campanula americana

We investigated the effect of petal-size variation in tall bellflower, Campanula americana, on pollen removal rates, seed set, and pollinator behavior. Pollen removal, seed set, and the behavior of two bee visitors (Bombus and Halictus) were assessed in flowers that had 0%, 50%, or 100% of their petal lobes removed. Pollen removal rates did not differ significantly among the three treatment groups, probably due to the presence of an ineffective (ugly) pollinator (Halictus), which quickly removed pollen from completely reduced flowers. Seed set was significantly positively correlated with petal size. Bumblebees discriminated against reduced flowers, thereby lowering pollen movement to reduced flowers. Halictid bees rarely contacted stigmatic surfaces and thus pollen delivery to reduced flowers was limited. The presence of an ugly pollinator probably obscured the effect of petal reduction on pollen receipt, thus masking the impact of attractive structures on male fitness. Given the positive relationship between seed set and petal size, we caution against ascribing expenditure on attractive structures exclusively to male function. In plants with many olules, expenditure on attractive structures may affect seed set.     

Floral sexual dimorphism and flower choice by pollinators in a nectarless monoecious vine Akebia quinata (Lardizabalaceae)

We investigated the pollination system and movement patterns of pollinators among flowers of the nectarless, monoecious vine Akebia quinata in natural populations and experimental floral arrays. Female flowers did not offer any rewards for pollinators and were larger than male flowers. Pollinators of A.quinata , such as small solitary bees and hoverflies, clearly discriminated between male and female flowers. Hoverflies always visited male flowers and rarely visited female flowers. In contrast, solitary bees tended to visit female flowers first when entering the array, but then switched to male flowers within the same foraging bout. This tendency disappeared when the sepal size of female flowers was experimentally reduced to the size of male flowers. Thus, observed non-randomness in flower choice by solitary bees may be caused by female>male sexual dimorphism and may increase the chance of cross-pollination and lower the probability of geitonogamous pollination in a single visit to a plant. Therefore, floral sexual dimorphism in A.quinata is considered to be an adaptation for deceptive pollination associated with discriminating pollinators.     

Floral sexuality and breeding system in gum karaya tree, Sterculia urens

Comprehensive studies were carried out on phenology, floral sexuality, pollination biology, pollen-pistil interaction, breeding system and fruit and seed set on three populations of gum karaya tree (Sterculia urens). The species is andromonoecious and produces a large number of male and a limited number of bisexual (functionally female) flowers. The numbers of male and bisexual flowers varies not only between trees but also during the flowering period within a tree. Each male flower produces about 5000 fertile pollen grains. Neither in morphology nor in number, is there any difference between pollen grains in the bisexual and male flowers. However, pollen grains of bisexual flowers are completely sterile and incapable of siring any seeds. Their anthers, however, serve to attract pollinators; the emasculated bisexual flowers fail to do so. Thus S. urens is apparently andromonoecious but exhibits cryptic monoecy. That the species is self-incompatible was confirmed by controlled pollinations. The self-incompatibility is of the late-acting type and manifests after the entry of the pollen tube into the ovule. Apis indica is the only pollinator recorded by us and wind plays no role in pollination. The efficacy of pollination is low as only 56% of flowers were estimated to be pollinated. The pollen load on one-third the number of pollinated stigmas was lower than the number of ovules present. Fruit set under open pollination is poor and is highly variable from tree to tree (0.7–3.2%). Apart from pollination constraint, limited resource availability may also contribute to low fruit set.     

The flower biology ofCephalanthera longifolia (Orchidaceae)—pollen imitation and facultative floral mimicry

Solitary bees (Halictus sp.) were found to be the effective pollinators ofCephalanthera longifolia. In the same foraging flight the bees also visit flowers ofCistus salviifolius which has a similar colour pattern.Cephalanthera offers no reward to its pollinators, but orange papillae on its labellum successfully imitate pollen ofCistus. AsCephalanthera also attracts pollinators in the absence ofCistus, this is regarded as facultative floral mimicry.     

Significance of flower exploding pollination on the reproduction of the Scotch broom, <Emphasis Type="Italic">Cytisus scoparius</Emphasis> (Leguminosae)

The significance of single flower visits by pollinators on the reproductive success of the Scotch broom, Cytisus scoparius, with explosive flowers was investigated. Unexploded flowers (intact flowers) produced no fruit, implying that autonomous selfing and apomixis do not occur and that the explosion of flowers by insects is necessary for the fertilization of flowers. The fruit set in the natural condition was intermediate between that in artificial selfing or exploding and that in artificial outcrossing. Seed set and the ratio of seeds to ovules did not differ significantly between the natural condition and artificial outcrossing. The fruit set by a single flower visit from Apis mellifera was similar to natural conditions. Apis mellifera was the most frequent visitor and the number of flowers tripped open per hour was the highest among all the flower visitors. Approximately 75% of pollen on a flower disappeared in a single flower visit by A.mellifera. The small amount of pollen remaining on already-exploded flowers appears to be the reason why A.mellifera do not frequently visit already-exploded flowers. The higher seed set by outcrossing was believed to contribute to the seed set in both natural conditions and single flower visits by A.mellifera, and did not differ between them. Therefore, at least in Japan, A.mellifera appears to be the most important pollinator for C.scoparius, resulting in a high outcrossing rate with each single flower visit.     

Diverse visitors with various pollinator importance and temporal change in the important pollinators of <Emphasis Type="Italic">Geranium thunbergii</Emphasis> (Geraniaceae)

Assessing pollinator importance of each floral visitor to a plant species is a key to understanding plant–pollinator interaction. The present study examined visitation frequency, pollination efficiency, and pollinator importance of the full range of floral visitors to Geranium thunbergii natural population, by measuring seed-set. During 2 years of observations, the flowers were visited by at least 45 insect species belonging to four orders. Among the main 22 visitor species, 11 species belonging to three orders (Hymenoptera, Diptera, and Lepidoptera) acted as the efficient pollinators. In both years, Hymenoptera, especially bees, was the most important pollinator to G.thunbergii. Thus, the flowers could be considered as bee-pollinated. However, the most important species were not constant between years. The study also documented that the efficient pollinators have larger body sizes. The dish-shaped floral morphology, taxonomically diverse pollinators, and temporal change in the most important pollinators indicate that G.thunbergii–pollinator interaction is a rather generalized system. The results suggest that casual observations of visitation, or even precise measurement of pollinator importance in a single season is insufficient to identify important pollinators.     

Ultrastructural investigations on Lycopersicum peruvianum pollen activation and pollen tube organization after self-and cross-pollination

No differences have been observed in vivo between Lycopersicum peruvianum compatible and incompatible pollen during activation and pollen tube emission and organization, that is until 4 h and 30 min after pollination. During pollen activation the main events are the setting free of rough endoplasmic reticulum (RER) cisterns which were stacked in the mature pollen, the increase in the number of polysomes, and a great activity of the dictyosomes. Immediately after germination of the vegetative nucleus and the generative cell move into the tube, the generative cell diviting to form the male gametes; the tube then becomes organized in four zones. This series of changes is similar to what has already been observed in vitro except that in vitro the generative cell remains undivided and the whole process from seeding to tube organization takes 3 h instead of 4 h and 30 min after pollination, as it does in vivo. Our findings are compatible with the main models of the tube inhibition mechanism proposed till now.Abbreviations RER rough endoplasmic reticulum - GC generative cell - VN vegetative nucleus - GP germinative pore Research performed under C.N.R. (Italian National Research Council) program Biology of Reproduction     

Are nectar-robbers mutualists or antagonists?

As exploiters of plant-pollinator mutualisms, nectar-robbers remove rewards (nectar) without providing pollination services. Though one might expect nectar-robbing to be costly to plants, it may instead benefit plants by indirectly increasing pollen dispersal. I investigated the direct effects of nectar-robbing bees (Xylocopa californica) on floral rewards and behaviors of pollinators visiting desert willow (Chilopsis linearis) and indirect effects of robbing on the reproductive success of the plant. Nectar-robbers reduced nectar; while unrobbed and robbed flowers were equally likely to contain nectar, nectar volumes were smaller in robbed flowers with nectar. Apis mellifera (honeybees), ineffective pollinators in terms of pollen deposition, avoided robbed flowers. In contrast, Bombus sonorus (bumblebees), effective pollinators, did not avoid robbed flowers. While bumblebees tended to spend less time in robbed flowers, the time that they spent in flowers was not correlated with pollen deposition. Using powder mimicking pollen, I found that on some days, powder was dispersed farther or to more flowers from robbed flowers, indicating that robbing may sometimes benefit plants by increasing male reproductive success. Powder movement suggested that the effect of robbing on male reproductive success ranged from costly to beneficial. The outcome for flowers that were marked early each morning was a function of prevalence of robbing and abundances of effective pollinators, but not a function of spatial variability among trees in prevalence of robbing or the abundance of ineffective honeybees. Unlike powder dispersal, female reproductive success, measured by fruit set and the number of pollen tubes growing in styles, was not affected by robbing. Thus, robbers did not reduce plants female reproductive success either directly by damaging flowers or indirectly by reducing pollen deposition by pollinators. Overall, this study indicates that nectar-robbers were not often costly to plants, and sometimes even benefited plants.     

“Ixoroid” secondary pollen presentation and pollination by small moths in the Malagasy treeletIxora platythyrsa (Rubiaceae)

The pollination biology ofIxora platythyrsa (Rubiaceae) was studied in NW. Madagascar. The plant displayed cream-yellow, nocturnally fragrant, nectariferous, tubular and strongly protandrous flowers. These had an ixoroid secondary pollen presentation mechanism: prior to anthesis, anthers exhausted their pollen onto unripe stylar heads. From this position pollen of male-stage flowers later adhered to primarily the probosces of small visiting nocturnal noctuid and geometrid moths. — Pollen was subsequently raked off moths' probosces by receptive, copiously papillose stigmas of female-stage flowers. Principal pollination adaptation was probably to the noctuid moth subfam.Sarrothripinae.     

Oviposition choices by a pre-dispersal seed predator (Hylemya sp.)

Summary Although the importance of pollinators has most often been examined in the evolution of floral characters, seed predators may also play a role in shaping floral evolution. In this study, I examined the role of interplant distance, plant size, and flower morphology on Ipomopsis aggregatás (Polemoniaceae) attractiveness to a pre-dispersal seed predator, Hylemya sp. (Anthomyiidae) and to hummingbird pollinators. The attractiveness of I. aggregata individuals to Hylemya was nonlinearly related to interplant distance in experimental arrays. Clumped and highly dispersed plants were preyed upon more frequently than those at intermediate distances. I found no relationship between interplant distance and visitation rates by hummingbird pollinators in these experimental arrays. However, in natural populations studied, clumped plants were more frequently approached by hummingbirds than those growing more widely dispersed. Display size was unrelated to visitation by Hylemya on inflorescences I clipped and maintained as large, small and control. Display size was also unrelated to the total number of visits by hummingbird pollinators to each of these experimental plants, however large display plants were more likely to be visited first in any given visitation sequence. Of various morphological measurements, corolla length showed the strongest positive correlation with Hylemya egg presence. To the extent that plant spacing and morphology is correlated with pollinator visits and ultimate seed set, Hylemya could be choosing flowers optimally, and playing a role in the evolution of floral traits.     

Suppression of heterotrimeric G-protein β-subunit affects anther shape,pollen development and inflorescence architecture in tobacco

The role of the heterotrimeric G-protein -subunit in plant development was studied in transgenic tobacco (Nicotiana tabacum L.) plants with reduced -subunit levels due to the antisense expression of the -subunit mRNA. The antisense plants had aberrant anther shape and produced non-germinating pollen. The anthers were sporadically transformed to petals, whereas other floral organs were not affected. The pollen grains were smaller than the wild-type pollen and had abnormal cell walls. The architecture of mature antisense plants was altered. The plants had long branched panicles and short stems. These data suggest that the -subunit of the plant heterotrimeric G-proteins is involved in the regulation of the reproductive phase of the tobacco life cycle, particularly in stamen development and pollen maturation.     

Pollen precedence and stigma closure: a mechanism of competition for pollination between Delphinium nelsonii and Ipomopsis aggregata

Summary Previous experiments showed that the sympatric herbs Delphinium nelsonii and Ipomopsis aggregata compete for hummingbird pollination and that deleterious effects of the former species on seed set of the latter involve interspecific pollen transfer. However, seed set was not reduced when pollen of both species was applied simultaneously to I. aggregata stigmas. Hence a competitive effect may require arrival of foreign pollen before conspecific pollen. To explore this possibility we subjected I. aggregata flowers to a competition treatment in which they received D. nelsonii pollen 6 h before I. aggregata pollen, or to a control in which they received only the conspecific pollen. Foreign pollen precedence decreased mean seed set by almost 50%, which is consistent with effects observed in previous experiments. Reduced seed set can be explained by the fact that foreign pollen often caused stigma lobes to close together within 1.5–6 h, reducing subsequent receptivity. Stigma closure was also elicited by conspecific pollen, but not by mechanical stimulation, and was influenced by size of the pollen load and identity of the plant being pollinated.     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Angiospermensippen derAlismataceae,Liliaceae, Juncaceae,Cyperaceae, Poaceae undAraceae

Some closely related members of the monocotyledonous familiesAlismataceae, Liliaceae, Juncaceae, Cyperaceae, Poaceae andAraceae with variable modes of pollination (insect- and wind-pollination) were studied in relation to the ultrastructure of pollenkitt and exine (amount, consistency and distribution of pollenkitt on the surface of pollen grains). The character syndromes of pollen cementing in entomophilous, anemophilous and intermediate (ambophilous or amphiphilous) monocotyledons are the same in principal as in dicotyledons. Comparing present with former results one can summarize: 1) The pollenkitt is always produced in the same manner by the anther tapetum in all angiosperm sub-classes. 2) The variable stickiness of entomophilous and anemophilous pollen always depends on the particular distribution and consistency of the pollenkitt, but not its amount on the pollen surface. 3) The mostly dry and powdery pollen of anemophilous plants always contains a variable amount of inactive pollenkitt in its exine cavities. 4) A step-by step change of the pollen cementing syndrome can be observed from entomophily towards anemophily. 5) From the omnipresence of pollenkitt in all wind-pollinated angiosperms studied one can conclude that the ancestors of anemophilous angiosperms probably have been zoophilous (i.e. entomophilous) throughout.

     

Bumble bee behavior and selection on flower size in the sky pilot,Polemonium viscosum

Summary In alpine Polemonium viscosum, plants having sweet-scented flowers are primarily pollinated by queens of the bumble bee species, Bombus kirbyellus. In this paper we ask whether two aspects of the pollination effectiveness of bumble bees, visitation rate and pollination efficiency, vary significantly with flower size in sweet-flowered P. viscosum.(i) Bumble bees visited plants with large flowers on 80–90% of encounters, but visited those with smaller flowers on only 49% of encounters. (ii) However, the gain in pollination that large-flowered plants obtained via increased visitation was countered in part because bumble bees deposited fewer outcross pollen grains per visit on stigmas of large flowers than on those of small ones. When both visitation rate and pollination efficiency are taken into account, the predicted value of a single bumble bee encounter declines from 1.06 seeds for flowers larger than 18 mm in diameter to 0.55 seeds for flowers smaller than 12 mm in diameter. Our results suggest that bumble bee pollinators of P. viscosum prefer flower morphologies that are poorly suited for precise pollination. Such behavioral complexities are likely to place constraints on the evolution of optimal floral design.     

Pollination biology of two species ofKielmeyera (Guttiferae) from Brazilian cerrado vegetation

The pollination biology and breeding systems ofKielmeyera coriacea andK. speciosa, two sympatric woody species common in the cerrado vegetation of C. Brazil, were studied. Both species have similar nectarless, polystemonous Papaver-type flowers which are visited by a similar spectrum of insects, though they bloom in different seasons and are thus phenologically isolated. Large carpenter bees seem to be the most important pollinators and these and other bees effect buzz pollen retrieval despite the fact that anthers are not poricidal. Both species ofKielmeyera possess strong xenogamous breeding systems. The presence of staminate flowers and andromonoecy inK. coriacea, as well as the longevity ofK. speciosa flowers are discussed as alternative strategies to improve pollination success and reproductive efficacy.     

Gene flow in an almond orchard

Summary Gene flow by pollen between trees is essential for nut set in commercial almond orchards, due to the self-incompatibility of almond cultivars used. A study of gene flow has been carried out in an orchard composed of single rows of a pollinating cultivar between every double row of the most commercially desirable cultivar, Nonpareil. This two-to-one planting pattern was repeated throughout the orchard, and several pollinating cultivars were used in various parts of the orchard in an attempt to provide flowers for cross-pollination with Nonpareil at all stages of flowering of the latter. Using isozyme markers GPI-2, LAP-1, AAT-1, PGM-1, and PGM-2 and three newly-defined isozyme markers for almond — IDH, G6PD, and SDH — it has been shown that the gene flow resulting in nut set is quite restricted, taking place most strongly between neighboring halves of cross-compatibile pairs of trees. Even that half of a tree facing away from the pollinating tree has significantly less gene flow to it, while the next tree further on has few nuts set by fertilization from the pollinating tree under consideration. This result is surprising considering the comparatively large distances that the honeybee brought into the orchard in large numbers must travel within the orchard. To explain this apparent paradox and the observation that in most cases only a small proportion (<20%) of flowers set nuts, it is suggested that the honeybee predominantly visits only one cultivar, flying along the row of the cultivar to do so, and that cross-pollination results from accidental or rare visits involving two or more compatible cultivars.     

Viability and sunlight sensitivity of oak pollen and its implications for pollen-mediated gene flow

Pollen-mediated gene flow and the male reproductive success of wind-pollinated trees depend on the initial viability of the pollen and the changes that occur in its viability during transport in the atmosphere. The viability of Quercus robur pollen was determined before and during exposure to sunlight by in vitro germination and the fluorescein diacetate reaction (FCR) in 2002 and 2003, respectively. These experiments allowed us to calculate initial pollen viability and pollen sensitivity to sunlight. The germination test revealed a lower initial pollen viability (25–65%) than the FCR (53–92%). Following 9.5 h of irradiation the viability was reduced to 75–100% as determined by the in vitro germination test or to 40–70% as determined by the FCR. The actual values of initial pollen viability and pollen sensitivity to sunlight were used to define a range of values for modelling pollen dispersal using the mesoscale meteorological model METRAS. The deposition patterns of viable pollen varied by as much as a factor of 14 by changing the viability parameters in the range of the observed values. This suggests significant differences in male reproductive success. Variations in initial pollen viability have stronger effects on the gene-flow pattern than do variations in pollen sensitivity to sunlight. In particular, pollen distribution throughout the local environment is shaped by the initial pollen viability, while pollen sensitivity to sunlight mainly influences long-distance pollen dispersal.This revised version was published online in February 2005 with corrections to an incorrect sentence in the Results section. Under Pollen Sensitivity it should read: decreased faster when assessed by the germination test than by the FCR.     

Comparative studies of pollen and fluorescent dye transport by bumble bees visiting Erythronium grandiflorum

Summary In the Colorado Rocky Mountains the glacier lily Erythronium grandiflorum exhibits a striking dimorphism in pollen color and is commonly pollinated by the bumble bee Bombus occidentalis. We induced bees to visit sequences of flowers in a flight cage, and compared dispersal of distinctively-colored pollen and fluorescent pigment (dye) that the bee had picked up at a single donor flower. Nonparametric and parametric analyses showed that dispersal properties of pollen and dye differed; consistently less pollen was deposited and it was carried consistently shorter distances than dye. Dye thus does not provide an accurate means of assessing exacty where or how far pollen travels in this plant-pollinator system. On the other hand, both pollen and dye responded similarly to several experimental manipulations of donor and recipient flowers. Hence dye may well be of value for a qualitative investigation of how floral traits influence pollen dispersal.     

Further attempts to induce “egg transformation” using irradiated pollen

Summary Numerous pollination treatments involving heavily irradiated (40–100 krad) pollen of diverse plant species failed to produce any clear cut egg transformants of the type reported by Pandey in Nicotiana. Genetic stocks of pea, rapeseed, and apple, bearing multiple Mendelian markers, were employed to detect any possible transformation events. For each plant species, an optimal level of irradiation was determined which would allow normal pollen tube growth leading to fertilization, but which would prevent the formation of normal hybrids due to the pulverized condition of the chromosomes contributed by the irradiated pollen. Pollination treatments included selfing, pollination with donor pollen mixed with self pollen, pollination with irradiated donor pollen mixed with self pollen, pollination with irradiated pollen followed by a delayed self pollination, and pollination with irradiated pollen by itself. None of these treatments produced clearly transformed seedlings. The total number of potential transformation events screened was in excess of 6,046 including 2,268 for pea, 3,309 for rapeseed, and 469 for apple. It is concluded that if egg transformation occurs outside of Nicotiana it is a rare event, and its frequent occurrence in Nicotiana must be, at best, an isolated phenomenon.Approved by the Director of the New York State Agricultural Experiment Station for publication as Journal Paper No. 3557     

Effects of temperature on phyllody expression and cytokinin content in floral organs of rose flowers

Formation of leaf-like organs known as phylloids in Rosahybrida cv. Motrea flowers was promoted by exposure of plants toelevated temperatures. At a day/night temperature regime of26°C/21°C respectively theproportion of malformed flowers exhibiting phyllody was four times higher thanthat in flowers of plants grown at21°C/15°C. The number ofpetals in phyllody-expressing flowers was higher than that in normal flowers.The total content of endogenous cytokinins in young flower buds of plantsexposed to the lower temperature was six times higher than that at the highertemperatures. The effects of the reduced temperature were pronounced on all thegroups of cytokinins examined. However, the proportion of the various cytokiningroups remained similar at both temperature regimes. In contrast to thecytokinins in the flower buds, the content of all cytokinin types in youngleaves increased following exposure to the higher temperature and was reducedbythe lower temperatures. After 11 weeks at the lower temperature, about18% of the flowers remained malformed, whereas at the higher temperatureabout 20% of the flowers still remained normal. All thephyllody-exhibiting flowers were formed on vigorously grown basal shootscharacteristic to Rosa hybrida plants, whereas the normalflowers at the elevated temperatures were formed on lateral shoots which weremost distal to the plant base. However, irrespective of the season, thepresenceof normal and malformed flowers was observed on plants kept growing at standardconditions of 30°C/17°C inthegreenhouse. This phenomenon led us to examine the cytokinins in floral organsofnormal and malformed cv. Motrea flowers grown in the greenhouse as well as inflowers of a complete rose mutant known as a 'Green Rose(Rosa chinensis viridiflora). The highest content ofcytokinins was found in the pistils and stamens of normal 'Motreaflowers. On the other hand, the content of cytokinins in leaf-like style-tubesin the malformed flowers as well as in partially malformed ovaries at the baseof phylloids was significantly lower. A low content of cytokinins was alsopresent in petals of both normal and phyllody-exhibiting flowers and the lowestcontent has been found in the phylloids of the 'Green Rose. Apossibility of mutant deviations in metabolism of cytokinins in rose plants isdiscussed.