THE MOTION OF WINDBORNE POLLEN GRAINS AROUND CONIFER OVULATE CONES: IMPLICATIONS ON WIND POLLINATION

Comparisons are presented between the three-dimensional airflow patterns created around and by a scale model of a conifer ovulate cone and the trajectories of windborne pollen grains around Picea, Larix, and Pinus ovulate cones. Three general components of the airflow pattern around an ovulate cone model are 1) doldrum-like eddies, rotating over the adaxial surfaces of cone scales and directed toward attached ovules, 2) airflow spiralling around the cone axis along cone scale orthostichies and parastichies, and 3) a complex pattern of vortices (“umbilicus”) directed toward the leeward surface of the ovulate cone. The observed trajectories of pollen grains around cones of Picea, Larix, and Pinus conform to two of these three airflow components: 1) pollen grains are seen to roll along cone scales toward the distal scale margin and to become reentrained in airflow directed backward toward attached ovules, and 2) pollen grains passing around the cone are deflected into the “umbilicus” airflow pattern, where they either settle on or impact with cone scales (approach trajectories), or where they approach the leeward cone surface but are deflected away by airflow passing under the cone (Z-shaped trajectories). Vectoral analyses of pollen grain motion reveal a complex pattern of trajectories influenced by boundary layer conditions defined by ovulate cone geometry and ambient airflow speed. Wind tunnel studies of ovulate cones subtended by leaves and stem indicate that leaves circumscribing the cone act as a snowfence, deflecting windborne pollen toward the cone. Vectoral analyses of airflow patterns and pollen grain trajectories close to ovulate cones indicate that wind pollination in conifers is a non-stochastic aerodynamic process influenced by cone-leaf morphology and the behavior of pollen grains as windborne particles.     

AIRFLOW PATTERNS AROUND SOME EARLY SEED PLANT OVULES AND CUPULES: IMPLICATIONS CONCERNING EFFICIENCY IN WIND POLLINATION

Aerodynamic analyses showing characteristic airflow patterns and the potential for wind-mediated pollination are presented for models of Paleozoic (Carboniferous) ovules and ovulate cupules (i.e., Genomosperma kidstoni, G. latens, Salpingostoma dasu, Physostoma elegans, Eurystoma angulare, and Stamnostoma huttonense). Lobes on ovules and cupules are shown to produce localized regions of turbulent flow with a concomitant reduction in airflow velocity. Data based upon models that mimic the characteristics of windborne pollen (= pseudopollen) show that these regions of turbulent flow correspond to those in which suspended pseudopollen impact with ovule and/or cupule surfaces. These data have bearing on a sequence of ovule morphologies purported to show the evolution of the integument by the progressive reduction in length of “preintegumentary” lobes and their acropetal fusion. As the preintegumentary lobes of the models studied consolidate around the megasporangium, regions of turbulent flow and high pseudopollen impact become localized around the pollen chamber or salpinx. The general morphologic trend envisioned for the evolution of the ovule is seen to be associated with an aerodynamic streamlining and an increased potential for wind-mediated pollination. Data for hair-bearing ovules and for ovulate cupules are discussed within the context of possible selective pressures favouring streamlining.     

AERODYNAMICS OF EPHEDRA TRIFURCA: I. POLLEN GRAIN VELOCITY FIELDS AROUND STEMS BEARING OVULES

The behavior of pollen grains within the airspace around Ephedra trifurca is described. Vectoral analyses of pollen grains moving around stems and ovules indicate a complex pattern of directional and magnitudinal changes in trajectories that can be related to the geometries of surfaces that obstruct airflow. Pollen grains, passing around cylindrical stems oriented normal to the direction of ambient airflow, are deflected in circumrotating non-laminar flow-patterns. Stems tilted downwind deflect pollen grains into trajectories along leeward surfaces of stems. These trajectories travel acropetally in a spiraling pattern and may intersect airflow patterns created around and by ovules. Computer analyses of pollen motion in the vicinity of ovules indicate that pollen vector-direction is highly canalized and directed toward micropyles. Within the immediate vicinity of micropyles, which produce pollination droplets, analyses indicate that the magnitudinal variance of pollen grain vectors is high (spanning three orders of magnitude). This variance coincides with dramatic changes in the local Reynolds numbers, resulting in a localized region around the micropyle in which neither viscous nor inertial forces predominate. Based on additional aerodynamics parameters (vector curl, vector-divergence, and vector curl-differential) it is shown that the region around the pollination-droplet is characteristically a “pollen sink” (pollen grains collect in this airspace) in which abrupt changes can occur in the angular momenta of airborne pollen grains. These aerodynamic analyses suggest that the morphology of ovules and the stems to which they are attached facilitates pollen capture by creating an aerodynamic “singularity” (= a unique region) around the pollination-droplet.     

AEROBIOLOGY AND POLLEN CAPTURE OF ORCHARD-GROWN PISTACIA VERA (ANACARDIACEAE)

The phenology of pollen release and pollen capture by Pistacia vera was studied in the field and laboratory respectively. Inflorescences of Pistacia vera were examined in a wind tunnel to determine whether the behavior of airborne conspecific pollen around receptive flowers differed as a result of changes in the shape and size of the inflorescence. In addition, the behavior of unclumped (single) and clumped pollen grains was studied to determine differences in the probability of their capture. Wind speeds within a commercial orchard during pollen shedding averaged 0.9–2.2 m/sec and atmospheric pollen concentrations were highest between 0900–1100 hr MST. Each of three stages in inflorescence development (defined on the basis of the number of exserted stigmas) was examined under identical ambient airflow conditions with equal concentrations of airborne pollen (1,000 grains/m³). The general pattern of pollen grain motion involves direct inertial collision by windward surfaces and by sedimentation of pollen onto leeward surfaces; clumped pollen rarely sedimented onto leeward surfaces. Small changes in ambient wind speed (0.5 m/sec to 1.0 m/sec) produced significant changes in the pattern of pollen motion around inflorescences and altered the number of pollen grains captured by leeward surfaces. Thus, wind pollination in P. vera is affected both by windspeed and by the shape or size of flower clusters. Differences in the behavior of clumped and unclumped pollen result from their inertial properties and responsiveness to local changes in the direction and speed of airflow. Unclumped pollen has a higher probability of being captured by leeward surfaces. The apparent insensitivity of pollen motion to differences in inflorescence size may ensure equitable pollination during the acropetal development of flowers.     

POLLEN CAPTURE AND WIND-INDUCED MOVEMENT OF COMPACT AND DIFFUSE GRASS PANICLES: IMPLICATIONS FOR POLLINATION EFFICIENCY

The motion induced by wind of Setaria geniculate (Lam.) Beauv. and Agrostis hiemalis (Walt.) B.S.P. panicles is shown to have harmonic characteristics that can influence pollination. During each cycle of its motion, the panicle of S. geniculata repeats a sequence of reorientations to the direction of ambient airflow. This repetition is correlated with a reiteration of airflow patterns through which airborne pollen grains must pass to reach floral surfaces. The panicle decelerates as it moves toward either of the two extreme positions of its oscillatory motion. The decelerations introduce a bias in the time the panicle spends in each of its orientations. This bias generates “preferred orientations” to the direction of ambient airflow which in turn results in biasing the aerodynamics of pollen capture. Similar phenomena are reported for the panicle of A. hiemalis, although a more complex pattern of oscillatory motion is observed. The reciprocity between the mechanics of oscillatory motion and the aerodynamics of pollen capture is shown to have deterministic properties which may surface above the “noise” produced by other, stochastic features influencing wind pollination.     

CONIFER OVULATE CONE MORPHOLOGY: IMPLICATIONS ON POLLEN IMPACTION PATTERNS

Empirically determined patterns of pollen impaction on the surfaces of pine ovulate cones are correlated with regions of nonlaminar flow created by the spatial arrangement and morphology (aspect ratios) of scale-bract complexes. Results from the serial discharge of pollen, upwind of ovulate cones, indicate that ovules on ovulate cones are preferentially impacted by pollen from their own species. Analyses indicate that while aerodynamic factors dominate the entrapment of pollen by ovulate cones, other factors such as pollen impaction-rebound and rebound-reentrainment are significant. Surface characteristics in addition to the settling velocities of pollen may play important roles in determining pollination efficiency. Wind tunnel analyses of the aerodynamic effects of scale-bract arrangement and aspect ratios indicate that each complex behaves as an aerofoil, deflecting air eddies toward the micropylar ends of ovules. The ovulate cone geometry, as a whole, deflects unidirectional wind into cyclonic vortices around the cone axis, each scale-bract deflecting nonimpacted pollen along orthostichies and parastichies. The morphology of the typical conifer ovulate cone is interpreted as a structure that optimizes anemophilous reproduction.     

EQUATIONS FOR THE MOTION OF AIRBORNE POLLEN GRAINS NEAR THE OVULATE ORGANS OF WIND-POLLINATED PLANTS

A technique is presented that is capable of predicting the motion of airborne pollen grains and the probability of pollen capture by wind-pollinated plants. Equations for the motion of rigid-walled particles (= pollen grains, spores, or Sephadex beads) in a supporting, compressible fluid (= air) are derived from the first principles of fluid dynamics. These equations are incorporated into a computer program (MODEL) which can be used with a desktop computer. The operation of MODEL requires empirical data on the pattern of airflow or the motion of a pollen species around the surfaces of the taxonomically relevant ovulate plant organ. With this information, MODEL can predict the behavior of any pollen species for which physical properties (size and density) are specified or empirically known. The significance of this procedure lies in the quantification of physical phenomena that influence the mechanics and fluid dynamics of pollen capture in wind pollination. The technique is illustrated and tested by its application to two grass species (Setaria geniculata and Agrostis hiemalis) for which velocity fields of pollen motion have been previously reported.     

THE AERODYNAMICS OF POLLEN CAPTURE IN TWO SYMPATRIC EPHEDRA SPECIES

Wind-tunnel analyses of the behavior of airborne pollen around ovules of two Ephedra species (E. trifurca and E. nevadensis) indicate that at certain airflow speeds (0.5 m/sec and 1.0 m/sec) each species is capable of biasing pollination in favor of conspecific pollen. A computer procedure was designed to evaluate the physical basis for this aerodynamic discrimination. This procedure indicates that differences in size and density confer significantly different inertial properties to the two pollen species. Operating within the specific aerodynamic environments generated around ovules from each species, these differences are sufficient to account for the biases observed in the probability of pollination. Within natural populations, there exists significant variation in pollen size (and possibly in density). Accordingly, it is possible that, under certain ambient wind conditions, ovules from each species can select subsets of the entire airborne population of Ephedra pollen.     

Effects of ovular secretions on pollen in Pseudotsuga menziesii (Pinaceae)

Effects of ovular secretions on pollen grains were examined in Pseudotsuga menziesii. The exine is cast off in the micropylar canal. A membranelike structure covers parts of pollen grains and appears to protect them. The outer intine consists of fibrous materials, but it also shows a thicker filamentous appearance in some ovules during pollen elongation. The inner intine is electron-dense. Its fibrous nature is occasionally visible. Dissolution of the outer intine varies in amount and manner in ovules from different trees. The plasma membrane near the pollen wall alternatively appears normal and distorted. These different morphologies of the outer intine and of the plasma membrane are considered to result from secretions from the ovule. The outer intine may contain electron-dense globules that are formed in the tube cell and traverse the inner intine. Pollen tube formation appears to be triggered by a secretion from the ovule. Cross-pollinated grains are less distorted compared with self-pollinated grains.     

The evolution of ovule number and flower size in wind-pollinated plants

In angiosperms, ovules are "packaged" within individual flowers, and an optimal strategy should occur depending on pollination and resource conditions. In animal-pollinated species, wide variation in ovule number per flower occurs, and this contrasts with wind-pollinated plants, where most species possess uniovulate flowers. This pattern is usually explained as an adaptive response to low pollen receipt in wind-pollinated species. Here, we develop a phenotypic model for the evolution of ovule number per flower that incorporates the aerodynamics of pollen capture and a fixed resource pool for provisioning of flowers, ovules, and seeds. Our results challenge the prevailing explanation for the association between uniovulate flowers and wind pollination. We demonstrate that when flowers are small and inexpensive, as they are in wind-pollinated species, ovule number should be minimized and lower than the average number of pollen tubes per style, even under stochastic pollination and fertilization regimes. The model predicts that plants benefit from producing many small inexpensive flowers, even though some flowers capture too few pollen grains to fertilize their ovules. Wind-pollinated plants with numerous flowers distributed throughout the inflorescence, each with a single ovule or a few ovules, sample more of the airstream, and this should maximize pollen capture and seed production.     

Variation of pollen and ovule parameters among different ploidy levels ofCorydalis (Fumariaceae)

The size and number of pollen grains and ovules are compared between 20 populations of different ploidy levels in two self-incompatible species ofCorydalis (Fumariaceae), to test the presence of ploidy-related variation in these reproductive characters. In both species, higher ploidy levels are associated with larger pollen grains, ovules and corolla, but the number of pollen grains and ovules are not different between ploidy levels. The investment per flower is consequently larger at higher ploidy levels, but the flower number per individual is lower, suggesting that the mode of partition of the investment for sexual reproduction varies between different ploidy levels. InC. orthoceras, sex allocation estimated by pollen:ovule ratios in number and volume is more female-biased in polyploids than in diploids. In spite of these variations, the characters studied can not be used as indicators of ploidy level due to the large overlaps between the ploidy levels.     

The timetable for development of maternal tissues sets the stage for male genomic selection in Betula pendula (Betilaceae)

The influence of the sequence of maternal tissue development in Betula pendula upon the potential for male gamete selection was investigated, and the timing of the fixed abortion of one of the two ovules was determined. We used scanning electronic microscopy, confocal laser scanning microscopy, and blue light microscopy. The stigmas remain fresh throughout male anthesis, and may also last after its end, depending on ambient temperatures. The presence of germinated pollen does not induce stigmatic necrotization, and grains may arrive at different times. The pollen tube tips remain within the stigma base until the end of female anthesis. The ovules will not develop until after necrotization of the stigmas. The pollen tubes thus have a fair start to the ovules, regardless of their different arrival times and of the original positions of the pollen grains at the stigma surface. Therefore, competition among different microgametophytes is possible, in spite of low pollination intensity. Our results indicate that when the first pollen tube penetrates an ovule, this ovule starts to outgrow the other one, and even if the other is also penetrated, its vascular support soon atrophies and the megagametophyte will shrivel. Fertilization of both ovules was never seen in this study.     

POLLEN-OVULE RATIOS AND BREEDING SYSTEM EVOLUTION IN SOLANUM SECTION BASARTHRUM (SOLANACEAE)

Pollen-ovule ratio, pollen grains per flower, pollen grain volume, pollen biomass, ovules per ovary, seeds per fruit, seed volume, seed biomass, and corolla size were studied to reveal patterns of variation within a monophyletic group. Solanum sect. Basarthrum includes 22 neotropical species of two clades, one having small, few-seeded red fruits, and the other having larger, many-seeded green fruits. The former includes self-incompatible species and a dioecious species, while the latter includes self-incompatible species, self-compatible (and autogamous) species, and a domesticate (the “pepino”). Although the pollen quantity and ovule quantity of the self-incompatible species are significantly higher in the green-fruited subgroup than in the red-fruited subgroup, the pollen-ovule ratios of the self-incompatible species are not significantly different between the two subgroups, suggesting parallel evolution of the pollen-ovule ratio. Furthermore, the pollen-ovule ratio tracks the breeding system: self-incompatible species have significantly higher pollen-ovule ratios than self-compatible species, resulting both from fewer pollen grains and more ovules of the latter. The pollen-ovule ratio of the dioecious species is among the highest of all, resulting only from fewer ovules. The pepino is self-compatible but has a pollen-ovule ratio like its wild self-incompatible progenitors and shows wide variation in seed production and pollen quality, but not in pollen quantity. Among all species, pollen size and quantity are inversely proportional, as are seed size and quantity.     

Pollination of Australian Macrozamia cycads (Zamiaceae): effectiveness and behavior of specialist vectors in a dependent mutualism

Complementary field and laboratory tests confirmed and quantified the pollination abilities of Tranes sp. weevils and Cycadothrips chadwicki thrips, specialist insects of their respective cycad hosts, Macrozamia machinii and M. lucida. No agamospermous seeds were produced when both wind and insects were excluded from female cones; and the exclusion of wind-vectored pollen alone did not eliminate seed set, because insects were able to reach the cone. Based on enclosure pollination tests, each weevil pollinates an average 26.2 ovules per cone and each thrips 2.4 ovules per cone. These pollinators visited similar numbers of ovules per cone in fluorescent dye tests that traced insect movement through cones. Fluorescent dye granules deposited by Cycadothrips were concentrated around the micropyle of each visited ovule, the site of pollen droplet release, where pollen must be deposited to achieve pollination. In contrast, Tranes weevils left dye scattered on different areas of each visited ovule, indicating that chance plays a greater role in this system. Each weevil and 25 thrips delivered 6.2 and 5.2 pollen grains, respectively, on average, to each visited ovule per cone, based on examination of dissected pollen canals. In sum, the pollination potential of 25 Cycadothrips approximates that of one Tranes weevil.     

POLLINATION,FERTILIZATION AND OVULE ABORTION IN OXALIS MAGNIFICA

Pollination, fertilization and ovule abortion were studied in Oxalis magnifica (Rose) Knuth, a strongly self-incompatible herb that regularly matures only a fraction of its ovules. Examination of cleared ovules indicated that among 9 individuals the average number of ovules fertilized ranged from 48–92%. The remaining ovules either failed to produce female gametophytes, or more commonly contained unfertilized female gametophytes, despite large numbers of compatible pollen grains that were placed on stigmas. Abortion of fertlized ovules could be detected first by the flattened and enlarged appearance of the endosperm nuclei, followed by visible deterioration of the embryo. Among individuals the rate of embryo abortion varied from 3.4–47.9%. At lower levels of pollination an almost one-to-one relationship existed between the number of pollen grains placed on stigmas and the number of seeds matured in the capsule. No threshold number of pollen grains necessary for successful pollen tube growth and fertilization could be demonstrated. Reduction in seed number through embryo abortion provides an opportunity for selection among developing seeds. The potential for this form of selection varies widely among individuals of Oxalis magnifica, which showed a 14-fold variation in the average percentage of aborted ovules.     

AERODYNAMICS OF WIND POLLINATION IN SIMMONDSIA CHINENSIS (LINK) SCHNEIDER

Wind tunnel analyses of Simmondsia chinensis (Link) Schneider or “jojoba” were conducted to quantify the behavior of airborne pollen grains around individual branches and leaves and near individual carpellate flowers. Field data (wind velocity) were used to ensure a correspondence between wind tunnel and natural conditions. Based upon the visualization of individual pollen grain trajectories, it is concluded that pollen deposition on stigmatic surfaces is influenced by large-scale aerodynamic patterns, generated by foliage leaves, and small-scale airflow patterns, formed around and by floral parts and stigmas. Leaves are seen to deflect airborne pollen grains into trajectories that can intersect ambient airflow at 90° angles, showering decumbent carpellate flowers with pollen. Similarly, flowers can deflect pollen upward and downwind, toward other flowers. The extent of floral bract and sepal recurvature is shown to influence the extent of pollen deposition by determining the characteristic airflow pattern around stigmas. Available evidence concerning the relatively recent evolutionary transition to anemophily in Simmondsia is interpreted within the context of morphological adaptations and exaptations favoring wind pollination.     

Pollen-ovule patterns in tribeTrifolieae (Leguminosae)

Relative pollen and ovule production in the genera of the legume tribeTrifolieae is explored particularly as to howMedicago with its explosive pollination mechanism compares with its allies.Medicago produces much larger, although much fewer pollen per ovule than the other five genera in the tribe; this is interpreted as a consequence of its highly specialized, irreversible pollination mechanism, which allows only one effective exchange of pollen with pollinators.Melilotus andTrifolium produce a comparatively large quantity of pollen; this can be related to their floral characteristics requiring abundant pollen to achieve fertilization. InMedicago andTrigonella, annuals convert a higher proportion of pollen grains and ovules into seeds than perennials.     

Pollen,ovules, and pollination in pea: Success,failure, and resilience in heat

Field pea (Pisum sativum), a major grain legume crop, is autogamous and adapted to temperate climates. The objectives of this study were to investigate effects of high temperature stress on stamen chemical composition, anther dehiscence, pollen viability, pollen interactions with pistil and ovules, and ovule growth and viability. Two cultivars (“CDC Golden” and “CDC Sage”) were exposed to 24/18°C (day/night) continually or to 35/18°C for 4 or 7 days. Heat stress altered stamen chemical composition, with lipid composition of “CDC Sage” being more stable compared with “CDC Golden.” Heat stress reduced pollen viability and the proportion of ovules that received a pollen tube. After 4 days at 35°C, pollen viability in flower buds decreased in “CDC Golden,” but not in “CDC Sage.” After 7 days, partial to full failure of anthers to dehisce resulted in subnormal pollen loads on stigmas. Although growth (ovule size) of fertilized ovules was stimulated by 35°C, heat stress tended to decrease ovule viability. Pollen appears susceptible to stress, but not many grains are needed for successful fertilization. Ovule fertilization and embryos are less susceptible to heat, but further research is warranted to link the exact degree of resilience to stress intensity.     

Regulation of LAT52 promoter activity during pollen tube growth through the pistil of Nicotiana alata.

The pollen-specific promoter of the LAT52 gene is known to direct expression of marker proteins during the last stages of pollen maturation and in very early pollen tube growth.We have examined the expression of LAT52-GUS during later stages of pollen tube growth in style and ovary of the relatively long-styled species Nicotiana alata. GUS activity was detected histochemically and found to be present in germinating pollen grains of N. alata and in tubes growing through the upper part of the style. No GUS activity was detected in 99% of the pollen tubes growing through the lower part of the style, but activity was present in tubes within the ovary. This finding indicates that the LAT52 promoter is regulated in growing pollen tubes, and is most active during the earliest and latest stages of pollen tube growth. GUS activity was also detected in some ovules, where it presumably marked the release of pollen tube cytoplasm into the ovule. The distribution of ovules with GUS activity within the ovary is not consistent with high-precision pollen tube guidance to the ovule. Received: 16 August 1999 / Revision accepted: 20 December 1999