Wind or insect pollination? Ambophily in a subtropical gymnosperm Gnetum parvifolium (Gnetales)

Gnetum (Gnetales) has long been regarded as insect pollinated due to its range‐restricted distribution in tropical rain forests, where wind pollination is supposed to be detrimental. However, ubiquitous pollen limitation in the tropics might cause transition to anemophily, or even ambophily for reproductive assurance, especially for gymnosperms such as Gnetum, which exhibit various anemophilous syndromes, including absence of petals, greenish color of strobili, dioecy and uniovulate flowers. Pollination treatments applied to Gnetum parvifolium in southern China revealed this rare and endangered species to be pollen limited and incapable of apomixis. Pollen grains of G. parvifolium did not adhere to each other and could be trapped by interception traps near the male and female strobili. Seed set in the netted treatment (anemophily) was significantly higher than in the bagged treatment (apomixis) but lower than open pollination (anemophily and entomophily), indicating that both wind and biotic pollination contributed to seed production of G. parvifolium. The occurrence of ambophily in Gnetum and the prevalence of anemophily in Ehpedra and Welwitschia suggest that wind may also play a role in the pollination of the ancient Gnetales.     

FLORAL BIOLOGY OF NEPENTHES GRACILIS (NEPENTHACEAE) IN SUMATRA

Nepenthes gracilis, a dioecious carnivorous plant, has inconspicuous flowers lacking petals. Nectaries distributed on the upper surface of four sepals secrete dilute nectar (3%–12% sugar concentration) at night, but the nectar immediately disappears during the day by evaporation in the sunny environment of Sumatra. Male flowers have a higher nectar production rate but lower sugar concentration of nectar than female flowers. Flowers of both sexes were visited by pyralid moths at night and by calliphorid flies in the evening. Pollen was found attached on these insects visiting Nepenthes flowers. The pattern of nectar production of sepals is regarded as attracting nocturnal flying insects and avoiding ants, while the pitchers attract ants by nectar secreted on the pitcher rim.     

Male-biased sex ratio and promiscuous pollination in the dioecious island treeLaurus azorica (Lauraceae)

Pollination ofLaurus azorica (Lauraceae), a dioecious Macaronesian tree, was studied. Male and female trees had the same size distribution. The population had 2.5 times as many male trees as females. In addition, males produced more flowers, and their inflorescences lasted longer. Individual flower lifetime and length of flowering season were the same in both sexes. Between the years of observation, one tree changed sex. Pollinators wereHalictinae bees and the flyTachina canariensis. The bees collected pollen and nectar and the fly collected nectar from both sexes. Both species visited other plants as well. The evolution of breeding systems inLauraceae is discussed.     

Anatomy,microstructure, and endogenous hormone changes in Gnetum parvifolium during anthesis

The study of the sexual systems of plants is very important for understanding their evolutionary process. Gnetum is a pivotal species in comprehending the seed plant phylogeny. However, the sexual systems of this genus are not fully understood. To investigate the sexual system of Gnetum, the anatomical structure, microstructure, and endogenous hormones were investigated from the differentiation of flower buds to fertilization in the floral organs of female and male of G. parvifolium (Warb.) C.Y. Cheng, widely distributed in the north of China. After 2 years of field investigations, the staminate strobili were observed on female plants for the first time and the abortive ovules were also found on male plants. In addition, high levels of gibberellin A₃ and zeatin riboside led to masculinization, whereas a high level of indole‐3‐acetic acid favored feminization. A comprehensive analysis of the results indicates that G. parvifolium may not be completely dioecious on morphology, but female and male plants can only perform their own gender function, which would provide a better understanding of the diversity of sexual systems in Gnetum, and reproductive structures of seed plants.     

Double fertilization in Gnetum gnemon (Gnetaceae): its bearing on the evolution of sexual reproduction within the Gnetales and the anthophyte clade

The fertilization process in Gnetum is critical to our understanding of the evolution of sexual reproduction within the Gnetales, a monophyletic group of nonfiowering seed plants that are the closest living relatives to flowering plants. Although much is known about the fertilization process in Ephedra, which is basal within the Gnetales, little is known about sexual reproduction in the derived sister groups Gnetum and Welwitschia. Ovules of Gnetum gnemon were collected at various stages after hand pollination and processed for light, fluorescence, and electron microscopy. Approximately 5 d after pollination, pollen tubes reach sexually mature female gametophytes, which are coenocytic. At that time, a binucleate sperm cell is found within each pollen tube. Within 7 d of pollination, double fertilization events occur when each of two sperm nuclei released from a pollen tube fuses with a separate, undifferentiated female nucleus within the free nuclear female gametophyte, which lacks differentiated egg cells. The products of double fertilization are two viable zygotes; endosperm is not formed. The lack of differentiated egg cells in Gnetum gnemon is unparalleled among land plants and the documentation of a regularly occurring process of double fertilization is congruent with the hypothesis that a rudimentary process of double fertilization evolved in a common ancestor of angiosperms and Gnetales.     

Pollination biology of a night‐flowering Galápagos endemic,Ipomoea habeliana (Convolvulaceae)

Ipomoea habeliana is an endemic, night‐flowering member of the Galápagos flora. Pollination experiments, flower‐visitor observations, nectar sampling, pollen transfer, and pollen to ovule ratio and pollen size studies were included in this project. The large, white flowers of this species set fruit via open pollination (55%), autonomous autogamy (51%), facilitated autogamy (91%), cross‐pollination (80%), diurnal open pollination (60%) and nocturnal open pollination (60%). Fruit set is pollen‐limited. Ants, beetles, crickets and hawk moths regularly visit the flowers. Ants are the most frequent visitors, but hawk moths are the only effective pollinators. Nectar is available throughout the night, but is most abundant early in the evening when hawk moth visits are most frequent. Experiments with fluorescent dust demonstrate intra‐ and inter‐plant pollen movement by hawk moths. Although this species is adapted for hawk moth pollination, it readily sets fruit via autonomous autogamy when no visits are made. Thus, it is concluded that it is facultatively xenogamous. Additional support for this conclusion is provided by the pollen to ovule ratio of 1407 and by the fact that the plants grow in a region that has few or no faithful pollinators. Conservation efforts for I. habeliana should include hand pollinations, which could significantly increase seed set. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 160 , 11–20.     

From near extinction to diversification by means of a shift in pollination mechanism in the gymnosperm relict Ephedra (Ephedraceae,Gnetales)

Pollination in gymnosperms is usually accomplished by means of wind, but some groups are insect‐pollinated. We show that wind and insect pollination occur in the morphologically uniform genus Ephedra (Gnetales). Based on field experiments over several years, we demonstrate distinct differences between two Ephedra species that grow in sympatry in Greece in pollen dispersal and clump formation, insect visitations and embryo formation when insects are denied access to cones. Ephedra distachya, nested in the core clade of Ephedra, is anemophilous, which is probably the prevailing state in Ephedra. Ephedra foeminea, sister to the remaining species of the genus, is entomophilous and pollinated by a range of diurnal and nocturnal insects. The generalist entomophilous system of E. foeminea, with distinct but infrequent insect visitations, is in many respects similar to that reported for Gnetum and Welwitschia and appears ancestral in Gnetales. The Ephedra lineage is well documented already from the Early Cretaceous, but the diversity declined dramatically during the Late Cretaceous, possibly to near extinction around the Cretaceous–Palaeogene boundary. The clade imbalance between insect‐ and wind‐pollinated lineages is larger than expected by chance and the shift in pollination mode may explain why Ephedra escaped extinction and began to diversify again.     

The Bizarre Inflorescence of Norantea brasiliensis (Marcgraviaceae): Visits of Hovering and Perching Birds1

The pollination biology of Norantea brasiliensis (Marcgraviaceae) was studied in the rain forest of southeastern Brazil. This plant presents bizarre, brush-type racemous inflorescences bearing numerous flowers and extrafloral cup-shaped nectaries. Flower anthesis is diurnal, nectar production is continuous and copious, and the sticky pollen is readily removed by visitors during the first morning hours. The ruby-coloured inflorescences were visited by eight species of hummingbirds (Trochilidae), and 10 species of passerine birds (three Coerebidae and seven Thraupidae). Hummingbirds hovered while probing for nectar and touched flowers occasionally, whereas passerine birds perched and made contact with flowers habitually. Due to differences in flower-visiting and general foraging behaviour, perching birds act as better pollen vectors than hovering birds. The inflorescence of Norantea brasiliensis seems well fitted for pollination by passerine birds, and the hexose-dominated nectar supports this idea. Pollination syndrome trends within Marcgraviaceae may stem from insect-pollinated, condensed and spike-like inflorescences which would give rise to bird-pollinated, brush-type inflorescences. From the same basic condensed inflorescence, bat-pollinated umbelliform inflorescence may be derived from bird-pollinated, pendulous and corymb-like inflorescences. These postulated inflorescence types are found among the extant species of Marcgraviaceae.     

Relative importance of diurnal and nocturnal pollinators for reproduction in the early spring flowering shrub Stachyurus praecox (Stachyuraceae)

Generalized pollination systems may be favored in early spring flowering plants, as during this period pollinator activity is unpredictable. Many previous studies have concentrated on the importance of diurnal visitors in pollination, and consequently, information on the contribution of nocturnal visitors to pollination in early spring is limited. This study was conducted to evaluate the relative importance of diurnal and nocturnal pollinators in the early spring flowering dioecious shrub Stachyurus praecox (Stachyuraceae), in two temperate forests in central Japan. Visitors to the female and male flowers were observed during day and night, and their relative contributions to seed set were compared. The pollinator observations revealed that the diurnal and nocturnal insects visited both male and female flowers, and that the main flower visitors were diurnal small bees and flies as well as nocturnal settling moths. The diurnal and nocturnal flower visitors also acted as pollinators, as the pollen grains of S. praecox were attached to the insects collected from the female flowers. Pollination experiments demonstrated that the contributions of diurnal pollinators to the seed set were higher than those of the nocturnal pollinators. The results of this study indicate that S. praecox has a generalized pollination system, comprising both diurnal insects and nocturnal settling moths. Although the roles of diurnal insects are more important in the pollination of S. praecox, nocturnal settling moths may have a complementary role in early spring.     

Moths pollinate four crops of Cucurbitaceae in Asia

Pollination ecology of many crops is not fully known, especially in tropical and subtropical regions. Non-bee pollinators may contribute substantially to crop yield, even if they do not receive much attention. Although moth pollination has fascinated ecologists and evolutionary biologists since Darwin, crop pollination by moths has not been well investigated and experimentally examined. Hence, we explored the pollination ecology of four cucurbit crops with crepuscular or nocturnal flowers. Lagenaria siceraria (Molina) Standl., Luffa acutangula (L.) Roxb., Trichosanthes anguina L., and Trichosanthes kirilowii Maxim. all display floral traits suggestive of moth pollination, such as opening around dusk or in evenings and secreting ample dilute nectar. We demonstrated that these crops’ flowers attracted a wide range of moth species, especially hawkmoths. The assemblage of flower-visiting moths varied according to location and season. Pollination treatments and pollen load analysis confirmed the pollination of the four crops by moths, especially hawkmoths. Our results provide evidence for the value as wild pollinators for the four crops, for which domesticated bees cannot provide reliable pollination services in practice. This study lends support to the proposal to pay more attention to the value of non-bee pollinators and to leave some areas unexploited in rural landscapes for the conservation of wild pollinators, including moths.     

Pollen tube growth in pistils of female-sterile and fertile plants ofOenothera mut.brevistylis

Summary Pollination and pollen tube growth was observed in pistils of fertile and female-sterile plants ofOenothera mut.brevistylis. The stigmas and styles of sterile plants were very irregularly formed and additional ovules often developed in the style. The malformations did not inhibit pollen tube growth but made it slower and less efficient. The pollen tubes grew in the ovary but did not find the ovules. Near their tips, pollen tubes branched or meandered without any clear direction. Lack of tropism caused by malfunction of the ovules is postulated in the female-sterile plant.     

Spatial fragrance patterns in flowers of Silene latifolia: Lilac compounds as olfactory nectar guides?

Floral odour can differ qualitatively and quantitatively between different parts of the flowers, and these spatial fragrance patterns within the flowers can be used by pollinators for orientation on flowers. Here we present results of spatial fragrance patterns within flowers of the dioecious Silene latifolia (Caryophyllaceae). Volatiles were collected and analysed using a highly sensitive dynamic headspace method, which allows dramatically reducing the sample time. From all flower parts, especially the petals and the anthophore emitted the typical flower volatiles of S. latifolia. However, compounds emitted from the petals differed from compounds emitted by the anthophore. The anthophore emitted the monoterpenoids lilac aldehydes and alcohols, whereas, all other typical scent compounds (e.g. benzoids, other monoterpenoids) were emitted by the petals. Lilac aldehydes are known to be behaviourally very attractive for noctuid moths, and they may serve as nectar guides in S. latifolia.     

Pollination biology and sexual differentiation ofOsyris alba (Santalaceae) in the Mediterranean region

Osyris alba L. is a widespread dioecious hemiparasitic shrub of S Europe, N Africa, and SW Asia. Male inflorescences are multiflowered whereas each female inflorescence is reduced to a single flower with persistent enlarged bracts. Pollination is a prerequisite for fruit and seed development and wind is unlikely to be an effective means of pollen spread. In southern Italy pollen is transported by small unspecialized flies and beetles. Both male and female flowers produce an indistinguishable sweet odour. Male flowers are produced in large numbers and over a larger period than the females and provide pollen, nectar, and staminal hairs as rewards for pollinators. The presence and function of staminal hairs with tip cells inOsyris alba has been reported for the first time. Female flowers are rewardless, producing neither mature pollen, nectar nor staminal hairs, but possess three modified yellow indehiscent anthers containing no viable pollen which may provide a strong visual feeding stimulus for pollinators. It is suggested that pollinators are attracted by deceit to female flowers by mimicry of the males and the floral mimicry is, therefore, intraspecific and intersexual. The floral characteristics and flowering phenology of male and female plants are consistent with this kind of mimicry. The female flower possesses a tricarpellary ovary with three ovules of which only one develops. The single seed, containing a small embryo and a large, rich endosperm, is borne in a red fleshy bird-dispersed fruit. The reduction in seed number per flower to one highly nutrient-invested seed, together with a reduction of the multiflowered inflorescence to a solitary flower and the sequential production of ripe fruits over an extended fruiting season, suggest that the female function is markedly resource-limited. It is suggested that, although all the reproductive characteristics present inOsyris alba, as well as hemiparasitism, had probably evolved before the end of the tropical Tertiary, they are of adaptive advantage in the nutrient and water-limited environment of the Mediterranean maquis.     

Pollination efficiency of cockroaches and other floral visitors of Clusia blattophila

• Cockroaches have rarely been documented as pollinators. In this paper we examine whether this is because they might be inefficient at pollination compared to other pollinators. Clusia blattophila, a dioecious shrub growing on isolated rocky outcrops in French Guiana, is pollinated by Amazonina platystylata cockroaches and provides a valuable system for the study of cockroach pollination efficiency.
• We examined the species composition of the visitor guild and visitation rates by means of camcorder recordings and visitor sampling. Then, we investigated the capacity for pollen transfer of principal visitors and found correlations between visitation rates and pollen loads on stigmas. In an exclusion experiment we determined the contributions of individual species to pollination success.
• Amazonina platystylata, crickets and two species of Diptera transferred pollen, but the number of transferred pollen grains was only related to visitation rates in the case of cockroaches. Crickets visited and rarely carried pollen. Dipterans were as frequent as cockroaches, carried similar pollen loads, but transferred much less pollen. An estimated 41% and 17% of ovules were pollinated by cockroaches and dipterans, respectively. The remaining ovules were not pollinated. There was no spatial variation in pollinator guild composition, but cockroaches visited flowers less frequently at the smaller study site.
• We demonstrate that cockroaches pollinate a large proportion of ovules. Their pollination service is not confined to one study site and, unlike that provided by dipterans, is not limited to certain years. We suggest that cockroach pollination has been overlooked and that cockroach‐pollinated plants, which share certain floral features, possess adaptations to pollination by cockroaches.

     

The Gnetales: Recent insights on their morphology,reproductive biology,chromosome numbers,biogeography, and divergence times

Ephedra, Gnetum, and Welwitschia constitute the gymnosperm order Gnetales of still unclear phylogenetic relationships within seed plants. Here we review progress over the past 10 years in our understanding of their species diversity, morphology, reproductive biology, chromosome numbers, and genome sizes, highlighting the unevenness in the sampling of species even for traits that can be studied in preserved material, such as pollen morphology. We include distribution maps and original illustrations of key features, and specify which species groups or geographic areas are undersampled.

     

Host plants and pollination regions for the long‐distance migratory noctuid moth,Hadula trifolii Hufnagel in China

Nocturnal moths are important pollinators of plants. The clover cutworm, Hadula trifolii, is a long‐distance migratory nocturnal moth. Although the larvae of H. trifolii are polyphagous pests of many cultivated crops in Asia and Europe, the plant species pollinated by the adult are unclear. Pollen species that were attached to individual migrating moths of H. trifolii were identified based on pollen morphology and DNA to determine their host plants, geographic origin, and pollination areas. The moths were collected on their seasonal migration pathway at a small island, namely Beihuang, in the center of the Bohai Sea of China during 2014 to 2018. Pollen was detected on 28.60% of the female moths and 29.02% of the male, mainly on the proboscis, rarely on compound eyes and antennae. At least 92 species of pollen from 42 plant families, mainly from Asteraceae, Amaranthaceae, and Pinaceae, distributed throughout China were found on the test moths. Migratory H. trifolii moths visited herbaceous plants more than woody plants. Pollen of Macadamina integrifolia or M. tetraphylla was found on moths early in the migratory season. These two species are distributed in Guangdong, Yunnan, and Taiwan provinces in China, indicating that migratory moths probably traveled about 2000 km from southern China to the Beihuang Island in northern China. Here, by identifying plant species using pollen, we gained a better understanding of the interactions between H. trifolii moths and a wide range of host plants in China. This work provides valuable and unique information on the geographical origin and pollination regions for H. trifolii moths.     

基于山地蜂蜜花粉组成的中华蜜蜂传粉多样性研究

中华蜜蜂为多种植物传粉,具有重要的生态服务价值。为进一步了解中华蜜蜂选择蜜粉源植物的多样性与特征,采用聚合酶链式反应(PCR)、DNA条形码、高通量测序等技术,研究了源自陕西宝鸡、贵州望谟和广西凤山蜂巢蜜中的花粉来源。通过对样品花粉rbcL基因片段扩增及测序、分析,发现各样本间可操作分类单元(OTU)水平上蜜粉源植物Alpha多样性指数无显著差异,同一地点样品不同重复间存在较高的相关性,而不同地区样本相关性较低。三地共鉴定出27目46科71属82种蜜粉源植物,其中宝鸡市、望谟县、凤山县样本中分别鉴定出57种、30种和40种,各地蜜粉源植物的物种组成存在较大差异。这些蜜粉源植物中,菊目植物种类最多,其次为毛茛目,分别有12种和7种。其中草本植物和木本植物(含乔木、灌木和木质藤本等)分别有44种和38种。研究对蜜粉源植物花序特点进行了分析,但并未发现相关性状对中华蜜蜂选择具有的影响。从属的区系成分分析,总体上温带分布区类型的植物属多于热带分布区类型的属,所占比例分别为46.5%和36.6%,但在凤山县样本中出现热带分布区类型多于温带分布区类型的情况。在不同样品中,红足蒿(Artemisia rubripes)、向日葵(Helianthus annuus)、芜青(Brassica rapa)、粗糠柴(Mallotus philippensis)、枇杷(Eriobotrya japonica)等分别占有一定优势,其与中华蜜蜂的关系值得进一步研究。根据蜜粉源植物的系统进化关系,中华蜜蜂访问的蜜粉源植物处于不同进化地位的分支,其中在菊类分支和蔷薇类分支比较集中。研究揭示了中华蜜蜂传粉植物组成特点,为进一步认知中华蜜蜂传粉服务功能的重要性提供依据。     

Reproductive consequences of mate quantity versus mate diversity in a wind-pollinated plant

Since most pollen travels limited distances in wind-pollinated plants, both the local quantity and diversity of mates may limit female reproductive success. Yet little evidence exists on their relative contribution, despite the importance of viable seed production to population dynamics.To study how variation in female reproductive success is affected by the quantity versus the diversity of surrounding mates contributing pollen, we integrated pollination experiments, data on natural seed set and seed viability, and AFLP genetic marker data in the wind-pollinated dioecious clonal forest herb Mercurialis perennis.Pollination experiments indicated weak quantitative pollen limitation effects on seed set. Among-population crosses showed reduced seed viability, suggesting outbreeding depression due to genetic divergence. Pollination with pollen from a single source did not negatively affect reproductive success. These findings were consistent with results of the survey of natural female reproductive success. Seed set decreased with the distance to males in a female plants’ local neighborhood, suggesting a shortage of pollen in isolated female plants, and increased with the degree of local genetic diversity. Spatial isolation to other populations and population size did not affect seed set. None of these variables were related to seed viability.We conclude that pollen movement in M. perennis is likely very limited. Both male proximity and the local degree of genetic diversity influenced female reproductive success.     

Synchronized dichogamy and dioecy in neotropicalLauraceae

In hermaphrodite neotropicalLauraceae a highly evolved dichogamous system is present which represents a kind of temporal dioecy. This system involves the existence of two flower morphs which are characterized by reciprocal phases of receptivity of the stigmas and pollen release. In some genera (Persea, Cinnamomum), nectar is produced as a reward for the flower visitors, while in other genera (Aniba, Clinostemon, Licaria), nectar is absent and pollen seems to be the only reward. This implies that in this case the flowers in the female stage must be deceptive flowers. In dioecious species of the generaOcotea andNectandra, both the male and female flowers attract the visitors with nectar. The pollen-ovule ratio of theLauraceae is comparatively low. — The type of reproductive system that characterizes theLauraceae, comprising functional dioecy, small, inconspicuously coloured flowers, pollination by small bees, and large, one-seeded fruits dispersed by birds, is quite prominent among trees of various families in the tropical lowland forest. The relationship between the different modes of flowering within theLauraceae and the causes for the correlation of their reproductive traits are discussed.     

The influence of nectar production on hawkmoth behavior,self pollination,and seed production in Mirabilis multiflora (Nyctaginaceae)

Nectar is the most common floral reward that plants produce to attract pollinators. To determine the effect of nectar production on hawkmoth behavior, pollen movement, and reproductive success in Mirabilis multiflora, I manipulated nectar volumes and observed the subsequent foraging behavior of the hawkmoth Hyles lineata and the resulting pollen movement patterns. Individual hawkmoths visited significantly more flowers on plants with more nectar. The increase in flower visits significantly increased pollen deposition on stigmas and pollen removal from anthers when nectar volume was raised to twice the highest level found in nature. As hawkmoths visited flowers consecutively on a plant, the proportion of self pollen deposited on stigmas increased significantly and rapidly. Based on simulated hawkmoth visits, seed set was significantly reduced for flowers later in a visit sequence. A simple model combining these results predicts that the form of selection on nectar production varies depending on pollinator abundance. Using a multiple regression analysis a nearly significant (P < 0.08) effect of stabilizing selection was detected during a single season as predicted by the model for the prevailing hawkmoth abundance. Although increased nectar production may indirectly affect plant fitness by reducing resources available for other plant functions, the direct effect of high nectar production on pollinator behavior and self pollination may generally limit floral nectar production.