Pollen heteromorphism is pervasive in <Emphasis Type="Italic">Thalictrum</Emphasis> (Ranunculaceae)

Among the angiosperms, features of pollen morphology such as grain size, aperture number and surface ornamentation display striking variation. It is less well appreciated that pollen morphology may vary within and among populations of the same species as well as within individual plants. In some species, individual plants produce multiple types of fertile pollen grains (called pollen heteromorphism). Aspects of pollen morphology, such as aperture number, are likely to affect fertilization success with different morphologies favored in different local competitive and ecological environments. This study surveys variation in pollen grain morphology among species throughout the genus Thalictrum. Pollen from individuals of 36 species was rehydrated from herbarium specimens, and light microscopy was used to quantify pollen grain aperture number and size. I find that pollen aperture-number heteromorphism is present within all Thalictrum species studied, and distributions of aperture-number morphs vary both within and among species. This study provides an example of significant pollen heteromorphism within a genus that also varies widely for pollination mode and sexual system.     

POLLEN APERTURE POLYMORPHISM AND GAMETOPHYTE PERFORMANCE IN VIOLA DIVERSIFOLIA

Pollen aperture polymorphism is studied in Viola diversifolia, where all plants produce three- and four-apertured pollen grains. We tested whether there are genetic differences among plants for the proportions of the different pollen morphs, and whether the morphs differ in gametophytic performance. Results show that the more apertures a pollen grain has, the more quickly it germinates but that few-apertured pollen grains have faster growing pollen tubes and longer life expectancies. The proportions of the different pollen morphs, together with pollen tube growth rates, may be inherited traits based on differences among maternal families. These results suggest that the different pollen morphs are favored in different pollination ecology situations. The production of several pollen morphs by the same individual could therefore be evolutionarily advantageous.     

Interspecific variation in pollen–ovule ratio is negatively correlated with pollen transfer efficiency in a natural community

The pollination efficiency hypothesis has long been proposed as an explanation for interspecific variation in pollen–ovule (P:O) ratios. However, no empirical study on P:O ratios has directly and quantitatively measured pollen transfer efficiency (PE). Here, we use a PE index, defined as the proportion of pollen grains removed from anthers that are subsequently deposited on conspecific stigmas, as a direct and quantitative measure of PE. We investigated P:O ratios, pollen removal and pollen deposition in 26 plant species in an alpine meadow, over three consecutive years. Our community survey showed that nearly 5% of removed pollen was successfully deposited on conspecific stigmas. The PE index ranged from 0.01% up to 78.56% among species, and correlated negatively with the P:O ratio across years. This correlation was not changed by controlling for phylogenetic relationships among species, suggesting that the interspecific variation in P:O ratios can be attributed to the probability of pollen grains reaching a stigma. The results indicate that the pollination efficiency hypothesis can help to explain interspecific variation in P:O ratios.     

Correlated variation in microtubule distribution, callose deposition during male post-meiotic cytokinesis, and pollen aperture number across Nicotiana species (Solanaceae)

In most flowering plants, a single cytokinesis follows the two meiotic divisions during pollen-grain ontogeny. Aperture pattern (i.e., aperture number and distribution on pollen surface) ontogeny could be linked to the processes ensuring the apportionment of the cytoplasm to the four microspores.This apportionment is achieved by radial arrays of microtubules organized around the nuclei. The cleavage planes are defined in the overlapping regions of opposing arrays extending from different nuclei. We followed the establishment of these arrays in two different lines of plants belonging to the genus Nicotiana that produce pollen grains with different aperture numbers. Different distributions of the microtubules have been observed, which can be interpreted as resulting from variation in the interactions between nuclei; these distributions appear to be correlated with aperture number.As a consequence, we propose that simultaneous cytokinesis allows the formation of multiple pollen morphologies. This mechanism is consistent with aperture number distribution within angiosperms and provides clues to help our understanding of the evolution of aperture number.     

Pollen morphology in relation to floral types and pollination syndromes in Pedicularis (Orobanchaceae)

Floral diversification in the genus Pedicularis (Orobanchaceae) is remarkable among flowering plants. In this genus, floral morphology and pollinator behavior are closely co-adaptive. In the current paper, pollen grains of 23 representative species of Pedicularis mainly from North America, with two species from Japan and two species from China, whose pollination ecology was previously studied, were examined using light microscopy and scanning electron microscopy. Two pollen aperture types and three kinds of exine ornamentation were recognized among these species. In addition, pollen data from previous and the current studies of Pedicularis were integrated and analyzed, together with some pollination characters. There was a significant association between pollen aperture types and corolla types, as well as between pollination syndromes and corolla types. However, there was no association of exine ornamentations with corolla types. The relationships and evolution of this genus were discussed with regards to pollen morphology, corolla types and pollination syndromes.     

Gametophytic vs. sporophytic control of pollen aperture number: A generational conflict

In flowering plants, the haploid phase is reduced to the pollen grain and embryo sac. These reproductive tissues (gametophytes) are actually distinct individuals that have a different genome from the plant (sporophyte), and are more or less independent. The morphology of pollen grains, particularly the openings permitting pollen tube germination (apertures), is crucial for determining the outcome of pollen competition. Many species of flowering plants simultaneously produce pollen grains with different aperture numbers in a single individual (heteromorphism). In this paper, we show that the heteromorphic pollen aperture pattern depends on the genetic control of pollen morphogenesis. This points out a conflict of interest between genes expressed in the sporophyte and genes expressed in the gametophyte. More generally, such a conflict should exist whenever heteromorphism is an ESS resulting from a bet-hedging strategy. For pollen aperture, heteromorphism has been observed in about 40% of angiosperm species, suggesting that conflicting situations are the rule. In this context, the sporo-gametophytic conflict could be one of the factors that led to the reduction of the haploid phase in plants.     

The cytological origin of the polyads and their significance in the reproductive biology of Mimosa bimucronata

The mechanism of polyad formation in Mimosa bimucronata (DC.) Kuntze is described and its significance in the reproductive biology is discussed. In this species c . 500 complexes of two coupled pollen mother cells (PMCs) per pollen sac are formed after the last mitotic division of the sporogenous tissue. All the pollen grains of each complex remain attached and form a bitetrad. This mechanism leads to the formation of many polyads per pollen sac. On this basis, we propose that in Mimosa the number of pollen grains per polyad (eight or 16) is determined by the number of cells that remain attached after the last mitotic divisions of the sporogenous tissue. Concerning the biological significance of the polyads in M. bimucronata , we find that the number of pollen grains per polyad is correlated with the number of ovules per ovary, and that the internal dimensions of the pit-shaped stigma are sufficient just for one polyad to be localized in it. Based on the number of polyads per flower this species has a high pollen/ovule ratio characteristic of xenogamous plants, in opposition to that expected for plants with composite pollen dispersal units. In contrast, it has low pollination efficiency, low fruit set, a 1/1 relationship between the pollen grains of a polyad and the number of ovules per ovary, and high seed set per fruit. These facts support the hypothesis that the occurrence of polyads in M. bimucronata would be an adaptation helping to ensure high seed set following a single cross pollination event.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 343–349.     

Comparison of two cacao (Theobroma cacao L.) clones for the effect of pollination intensity on fruit set and seed content

We compared the influence of pollination intensity (PI) on fruit set and seed number per pod in two cacao (Theobroma cacao L.) clones, IFC5 (Forastero Lower-Amazon Amelonado), which is self-compatible and known to produce a high number of seeds per pod under open pollination, and SCA6 (Forastero Upper-Amazon), which is self-incompatible and known to produce a low number of seeds per pod under open pollination. With both clones, PI had a positive effect on fruit set, with the maximum rate requiring more than 150 pollen grains per pod. One-half of the maximum rate of fruit set was reached with 39 pollen grains per pod for SCA6, and 78 for IFC5. With SCA6, a significant positive effect of PI on seed number per pod was also observed, with maximum seed number requiring more than 200 pollen grains per pod. In contrast, seed numbers with IFC5 were approximately equal over the PI range 100–800 pollen grains per pod. Patterns of seed number per pod were compared after: (1) open insect pollination, (2) low-intensity hand pollination and (3) high-intensity hand pollination. The patterns obtained with IFC5 under open pollination showed a peak of 40–50 seeds per pod, whereas the distribution patterns were more even with SCA6. The pattern of seed number under open pollination was similar to that obtained with high-intensity hand pollination for IFC5, and with low-intensity hand pollination for SCA6. We concluded that the high number of seeds per pod observed with IFC5 under natural insect pollination may be explained by a high number of compatible self-pollen grains on the stigma and by a severe drop of low-pollinated flowers eliminating potential lowfilled fruits. With SCA6, however, the number of compatible pollen grains deposited on the stigma was probably low under open pollination, and the flowers required lower pollen quantities to set fruit, which resulted in a high frequency of low seed numbers per fruit. This difference in the capacity to set low-seeded fruits might be considered as an adaptive trait related to the mode of reproduction.     

The effects of landscape on bumblebees to ensure crop pollination in the highland agricultural ecosystems in China

Honey bees and wild bees provide critical pollination services to agricultural ecosystems; however, the relative contributions of different bee taxa are not well understood. The natural habitats surrounding farmland support food and nesting resources for wild bees and therefore play an important role in the maintenance of crop pollination. In this study, we selected Cucurbita pepo L. (squash) as a model crop to investigate the relative importance of honey bees and bumblebees in pollinating the crop. Thirteen fields, which were surrounded by a gradient of natural habitat, were investigated on the Yunnan‐Guizhou Plateau in China. We measured the visit densities of honey bees and bumblebees, the number of pollen grains deposited in a single visit by the two bee taxa, as well as the overall pollen grains deposited on stigmas during a flowering day, and then used Bayesian inference to decouple the pollen grains deposited by either the honey bees or the bumblebees. Compared with honey bees, bumblebees deposited a higher number of pollen grains on stigmas in a single visit, but had a lower visit density than honey bees. Meanwhile, the bumblebee visit density increased along the proportion of natural habitat, while the honey bee visit density was not affected by the surrounding natural habitat. Data simulations using Bayesian inference showed that on a flowering day, the number of pollen grains deposited by bumblebees increased with the proportion of natural habitat in the surrounding landscape, but the number of pollen grains deposited by honey bees did not. Moreover, the total numbers of pollen grains deposited by honey bees or bumblebees alone were all below 2000 (the critical level to satisfy the pollination requirement of this crop). Pollen calculations demonstrated that the number of pollen grains deposited by the two bee taxa was greater than 2000 in fields surrounded by more than 13% natural habitat (grasslands and forests). The results revealed that bumblebees ensured C. pepo pollination in combination with honey bees in the highland agricultural ecosystems.     

新疆石竹属8种植物的花粉形态

利用扫描电镜对8种石竹属（Dianthus）植物的花粉形态进行了观察。结果表明:本属植物花粉为单粒花粉,圆球形,大小为30.5~44.0 μm,具多个萌发孔,孔数为12~16个,分散,孔圆形,孔径4.1~7.0 μm,具孔膜,孔膜上着生有若干棵小刺,花粉外壁具刺状和穴状纹饰,表面有或无穿孔。研究表明：花粉大小、表面纹饰、特别是萌发孔的形态特征存在较明显差异。可为石竹属植物的分类及野生资源利用等相关研究提供了可靠的孢粉学参考。     

野菊与菊花杂交中花粉活力和柱头可授性及胚胎发育研究

应用石蜡制片、活体压片、光学显微镜及扫描电子显微镜观察等方法,研究了四倍体河南云台山野菊(Dendranthema indicum)与栽培菊花'钟山金山'(D.grandiflorum 'Zhongshanjinshan')种间杂交中父本花粉活力、花粉在柱头萌发、花粉管生长及胚胎发育情况等.结果发现,父本云台山野菊的花粉活力在授粉时为12%左右.人工授粉后的不同时间,在柱头上都观察到正常萌发的花粉粒,且花粉管都能进入柱头,其中,在授粉后0.5 h时,平均每柱头有5.9粒花粉萌发;12 h时,为59.9粒;而24和48 h时,则分别降为47.1和35.7粒.此外,在授粉后8、10、12和15 d时,分别在49.1%、40.8%、39.7%和38.5%子房内观察到正常发育的胚胎,最终杂交结实率为44.8%,而母本自然开放结实率为52.3%.研究表明,授粉前其多数母本雌蕊发育良好、授粉后多数花粉能在柱头正常萌发和花粉管正常生长,在受精后大部分胚胎发育正常是野菊与栽培菊种间杂交较高结实率的重要保证,而授粉前父本较低的花粉活力对杂交结实率影响不大.     

Studies in Montia L. and Claytonia L. and Allied Genera

The pollen morphology of several genera in Portulacaceae is described. Particular attention has been paid to the genera of the subfamily Montioideae, as a stage of continued monographical studies. Among genera especially dealt with are Claytonia, Montia, Crunocallis, Naiocrene, Neopaxia, Mona, Maxia, Limnalsine, and Montiastrum. In the taxonomical treatment of these genera the pollen morphology has proved to afford many important additional characters.

The pollen grains of Claytonia are distinguished from those of the remainder in being 3-colpate. The grains of the Claytonia-type have many similarities with those of Lewisia, a genus of the subfamily Portulacoideae. The other genera of Montioideae have pantocolpate pollen grains. Among these genera several different pollen types are distinguished, chiefly with regard to the sexine structures and the aperture membranes. The Montiastrum-type is especially interesting, with tholate grains, a particular pollen type not met with in any other genus in the family. The pollen morphology of some genera in the Portulacoideae is also treated. In some species in Calandrina and Talinum pantotreme pollen grains are observed with apertures transitional between pori and colpi. The apertures of the pantotreme grains are arranged in characteristic patterns.

Particular attention has been given to the variation of the pollen morphological characters. This variation has been examined with regard to the differences between different populations of the same species as well as between different species. The greatest variation has been observed in the shape and size of the grains. The structure and sculpture and thickness of the sexine and the aperture membranes are less variable. Some polyploid taxa are connected with the occurrence of pollen grains with divergent and varying aperture numbers.

In a survey of the genera the taxonomical results of the investigation are presented with particular regard to the pollen morphology. The new genus, Maxia Ö. Nilss., is described. One new species, Montia clara Ö. Nilss., is described and some new combinations are made.

Pollen morphological diagnoses are given for 46 different taxa. The aperture conditions for 96 different species are presented.     

Wettable and unsinkable: the hydrodynamics of saccate pollen grains in relation to the pollination mechanism in the two New Zealand species of Prumnopitys Phil. (Podocarpaceae)

The pollination mechanism of most genera of the Podocarpaceae involves inverted ovules, a pollination drop and bisaccate pollen grains. Saccate grains have sometimes been referred to as 'non-wettable' due to their buoyant properties, while non-saccate pollen grains have been described as 'wettable'. The hydrodynamic properties of saccate pollen grains of seven podocarp species in five genera, Dacrydium Sol. ex G. Forst., Dacrycarpus (Endl.) de Laub., Manoao Molloy, Podocarpus L'Hér. ex Pers. and Prumnopitys Phil. have been tested in water, together with saccate and non-saccate pollen of four other conifer genera, Cedrus Trew (Pinaceae), Cephalotaxus Siebold & Zucc. ex Endl. (Cephalotaxaceae), Cupressus L. (Cupressaceae) and Phyllocladus Rich. ex Mirb. (Phyllocladaceae), and spores of three fern species and one lycopod species. All four spore types studied were non-wettable, whereas the bisaccate and trisaccate pollen types, like all other conifer pollen types, were wettable, enabling the grains to cross the surface tension barrier of water. Once past this barrier, grain behaviour was governed by presence or absence of sacci. Non-saccate and vestigially saccate grains sank, whereas saccate grains behaved like air bubbles, floating up to the highest point. In addition, the grains were observed to float in water with sacci uppermost, consistent with the suggestion that distally placed sacci serve to orientate the germinal furrow of the pollen grain towards the nucellus of an inverted ovule. Observations of pollen grains in the pollen chambers of naturally pollinated Prumnopitvs ovules confirmed this. The combination of buoyancy and wettability in saccate pollen has implications for the efficiency of the typical podocarp pollination mechanism.     

Pollen size in Carex: The effect of different chemical treatments and mounting media

Pollen size and pollen aperture size for ten species of the genus Carex L., native to Estonia, have been measured using light microscopy. The species selected represent different sections of the genus, a range of habitats and different chromosome numbers. The effects of two basic chemical treatments, two mounting media and the effect of chemically induced dehydration with tertiary butyl alcohol (TBA) on the size of pollen grains were then recorded.

In general pollen size and pollen aperture size of the species examined is highly variable at both intraspecific and interspecific levels. Carex hirta has notably larger pollen grains than any of the other species investigated and, although correlations between size and chromosome number in the species examined are limited, it also has the highest chromosome number. Statistically significant size differences resulted from variations in chemical treatment, mounting media and tertiary butyl alcohol (TBA) induced dehydration. Acetolysed pollen grains are larger than potassium hydroxide (KOH) treated pollen grains. Pollen grains dehydrated after chemical treatment with TBA are larger than pollen grains not dehydrated. Pollen grains mounted in silicon oil are smaller than grains mounted in glycerine. But considering the great size variation of Carex pollen grains, the size changes caused by preparation procedures fall within the size variation range of the species examined.

All the samples contained a high number of deformed pollen grains and pollen grains with hardly distinguishable or no lateral apertures.     

Zur Pollenmorphologie der Gattung Waltheria L. (Sterculiaceae)

The pollen grains of 69 specimens representing 44 species or supposed species of the genus Waltheria L. have been studied. Based mainly on differences in exine characters 2 pollen types, a spinulose and a suprareticulate one were pointed out. Both types show ± parallel trends in the development of the aperture system to an increase of aperture number, to a shortening of the colpi and to more distinct bounds and nexine thickenings of the ora. They exemplify important steps of aperture evolution, which in the spinulose type leads from zonocolporate by way of zonopororate to pantopororate and in the suprareticulate type from zonocolporate to pantocolporate grains. The tendencies of sporoderm evolution concern a more distinct morphological and partially also a regional differentiation of the exine elements (bacula, spinulae), which prove to be close correlated with the evolution levels of the apertures. According to the pollen morphological point of view the species of the genus Waltheria are to be arranged in two groups, which may be regarded as two related, but relative independent phyletic entities. These evidences speak in favour of a taxonomic     

Aerodynamics of Ephedra trifurca

Computer simulations are used to predict the behavior of pollen grains with different physical properties within the acceleration field created around the ovules of the gymnosperm Ephedra trifurca. A modelling procedure is given that (1) calculates the number of pollen grains captured by an ovule's pollination-droplet and (2) gives a correlation between pollination efficiency and the physical properties (= mass, size) of different types of pollen. Based on this procedure, the number of Ephedra pollen grains captured by micropyles can be less than the number captured from other species. However, the mass and size of Ephedra pollen grains appear to coincide with those predicted to yield a local maximum of pollination efficiency, i.e. slightly larger or smaller values of either mass or size would decrease the probability of capture. In addition, the properties of Ephedra pollen grains operate synergistically in the aerodynamic environment around ovules and are focused to collide with pollination-droplets. By analogy, the properties of Ephedra pollen coincide with those predicted for a localized adaptive peak. The physical properties of pollen grain types other than E. trifurca that can maximize pollen capture are not generally represented in the aerobiology of Ephedra during the pollination season. Therefore, the phenology of pollen release, community taxonomic-composition, and the physics of particle capture play collectively important roles in the reproductive success of Ephedra trifurca.     

An improved method for pollen release from anther: a case study with mangroves

The estimation of pollen production is widely used in pollination and reproductive ecology, aerobiology and pollen-vegetation relationship studies. Pollen release is the key step in estimating pollen production and therefore, the technique used to release pollen is critical. Three methods, the drying method (DM), mechanical method (MM) and chemical method (CM), are used to determine the amount of pollen released from anthers. Few studies have compared the results obtained from each method, making it difficult to determine which method is most appropriate in a given situation. In this study, we compared existing methods with a new method that increases the amount of pollen released from anthers. Eight species of mangrove trees from the Neilingding-Futian National Nature Reserve were selected for study. We combined the MM and CM to produce a new method (the ultrasonic method, UM). To determine the best ultrasonic treatment time using the UM, different durations were tested and the various responses were analysed. The relationships between pollen rupture and the ultrasonic treatment time, pollen size, exine thickness and aperture size were analysed. Finally, four methods for pollen release from anthers were compared. The results indicated that the UM could be an efficient method to release pollen from the anther. To avoid pollen rupture, ultrasonic treatment times in the UM should be less than 30 s. The pollen rupture rate was significantly correlated with the ultrasonic treatment time (r = 0.618, p < 0.001) and the aperture size (r = 0.248, p = 0.036). This comparative experiment indicated that the UM can generate a greater pollen yield than other methods while causing less pollen rupture than the MM. Therefore, the UM may be suitable for more plant species and more accurate for pollen production estimates than the existing three methods, particularly when estimating pollen production in all community plants.     

Pollen morphological evolution in bat pollinated plants

This study assesses to what extent bat pollination has acted as a selective force on pollen morphology. Earlier pollen studies have suggested convergent evolution of verrucate exine ornamentation. Furthermore pollen of bat pollinated plants has been reported to be bigger than that of plants pollinated by other means. The generality of these ideas is tested using a sample of 130 species of both bat pollinated plants and relatives with another mode of pollination. An analysis of pollen size, shape, aperture number and type, and ornamentation type of 35 plant groups in which a transition towards bat pollination occurred was performed and showed a significant effect for pollen size only. Bat pollinated plants have bigger pollen than their relatives. Pollen size was shown to correlate with style length. Pollen shape, aperture system and exine ornamentation are in general not very different in bat pollinated plants in comparison with their relatives. There is no consistent trend for rougher exines to be associated with bat pollination.     

The pollination biology of two species of Helicteres (Malvaceae) with different mechanisms of pollen deposition

Species of Helicteres are pollinated mainly by hummingbirds and bats. Most species pollinated by hummingbirds have a mechanism of depositing pollen on the top of the pollinator's head such as is shown in this work for Helicteres sacarolha. H. brevispira has an unusual mechanism of depositing pollen grains under the tail or on the abdomen of the hummingbirds. The top of the birds’ head may be considered an efficient place to transport pollen grains, because it is plain, easily accessible for deposition and donation of pollen and not easily accessible for grooming, while the tail is movable and inclined. Thus, H. brevispira pollinators may carry or transfer fewer amounts of pollen grains than H. sacarolha pollinators from one flower to another. If a large amount of pollen grains is lost, a higher quantity of flower or pollen has to be produced to guarantee reproductive success. Plants of H. brevispira set higher number of flowers and pollen grains per flower than H. sacarolha and have also higher rates of fruit and flower abortion. Thus, pollination efficiency of H. brevispira may be reached by high pollen and flower production. Flower change mechanism presented in this species may be also involved with the optimization of pollinator feeding and pollination efficiency.