Autonomous self‐pollination in the nectarless orchid Pogonia minor

The pollination biology of the nectarless orchid Pogonia minor was investigated in central Japan. The investigation revealed that the solitary flowers failed to attract pollinators, while high rates of fruit set were observed in the natural population. Comparable levels of fruit set were obtained in bagged, artificial self‐pollinated and artificial cross‐pollinated plants, indicating that the species is not pollinator‐limited for fruit set under natural conditions. Autonomous self‐pollination in P. minor resulted from a reduced rostellum, which allowed contact between the pollinia and the stigma. Self‐pollination is thought to be an adaptive response that provides reproductive assurance under conditions of pollinator limitation. Since pollen limitation is generally known to be frequent among deceptive orchids, strong pollen limitation is probably a driving force in the autonomous self‐pollination mechanism in the nectarless orchid P. minor.     

Biotic versus abiotic pollination in Oeceoclades maculata (Lindl.) Lindl. (Orchidaceae)

Geographical variation in the reproductive biology of widespread species often occurs at their distributional boundaries. We sought to determine whether such variation has occurred in an invasive orchid, Oeceoclades maculata, across its naturalized range. We compared its reproductive biology in a Brazilian population with that published for a population on the Caribbean island of Puerto Rico. In the state of São Paulo, O. maculata flowers between December and February, at the height of the rainy season. Similar fruit sets were observed in manual self (76%) and cross (70.4%) pollination treatments. The fruit set of plants protected from both pollinators and rainfall was 6.1%, whereas plants exposed only to rainfall had a fruit set of 41.4%, slightly less than the controls (48.3%). Like the Puerto Rico population, reproduction is primarily through rain‐assisted autogamy, but unlike observations made on the island, outcrossing can eventually occur. We observed two butterfly species (Heliconius ethilla narcaea and Heliconius erato phyllis) pollinating O. maculata. Secretory epidermal cells and trichomes of the spur lumen produced 0.7 µL of 25% (sucrose equivalents) nectar per flower each morning, which was stored in a dilated basal portion of the spur and reabsorbed by the afternoon. Thus, geographical variation in reproductive biology exists across the broad invasive range of O. maculata.     

Evolutionary disruption in the pollination system of Vanilla (Orchidaceae)

• Pollination of the pantropical Vanilla has been linked to melittophily and food deception.
• Here we investigated the role of flower traits on the reproduction of Neotropical Vanilla. We also studied the evolution of pollination systems in order to understand the origin of production of flower resources and the diversification of pollinators in this orchid genus. Our study was founded on data of adaptations in flower morphology, production of resources, scent release, pollinators and breeding systems of Vanilla and presenting new data on reproductive biology of V. palmarum. Data on reproductive biology of Vanilla were mapped onto a phylogeny to address our queries on the evolution of pollination systems in this genus.
• Vanilla palmarum shows a mixed mating system, with its facultative autogamous flowers being pollinated by hummingbirds. Its yellow flowers are scentless and produces nectar. Mapping of the pollination system onto trees resulted in one origin for bird pollination and at least two origins for autogamy in Vanilla. Nectar secretion has a single origin in the Neotropical thick-leafed lineage.
• Bird pollination of Vanilla is shown for the first time. The origin of ornithophily within a bee-pollinated clade is supported by flower morphology. Floral transitions to ornithophily have been favoured by the occupation of a distinct niche from that of the other thick-leafed Vanilla species. Despite its specialized pollination, V. palmarum is autogamous. A mixed mating system can promote reproductive assurance in the case of a decline in pollinator populations, or in areas where pollinator services are irregular or absent.

     

Reproductive biology and pollination of Utricularia reniformis A.St.‐Hil. (Lentibulariaceae)

Utricularia reniformis is an endemic Brazilian carnivorous plant, most common in high‐altitude grasslands. Knowledge of the reproductive biology of U. reniformis is essential for planning conservation strategies, but it is currently poorly understood. Thus, we studied the floral morphology, floral biology, breeding system and pollination of this species. U. reniformis produces and stores nectar in the flower spur, a classic feature of bee‐pollinated flowers, and we recorded Xylocopa sp. and Bombus sp. as pollinators. Moreover, although it is self‐compatible it is an obligate animal‐pollinated species, as the sensitive stigma avoids self‐pollination. However, in natural conditions reproductive success is low due to the rarity of visits from pollinators. We suggest that the low reproductive success caused by a deficit of pollinators may affect gene flow, causing loss of genetic diversity in U. reniformis.     

Delayed autonomous self-pollination in Hibiscus laevis (Malvaceae)

Delayed autonomous self-pollination allows outcrossing to occur while also ensuring that seeds are produced in the absence of pollen vectors. We investigated variation in the efficacy of this pollination mechanism in populations of Hibiscus laevis. Recurvature of stylar branches occurred after 1 d of anthesis, and in plants from Ohio, Illinois, Arkansas, and Oklahoma this behavior resulted in autonomous selfing (surprisingly, stylar movement was facultative in that it did not take place when the stigmas were already pollinated). In contrast to these more northern populations, the distance between anthers and stigmas was too great to allow autonomous selfing in plants from Texas, Mississippi, and Louisiana. Greenhouse studies of plants from Ohio demonstrated that autonomous selfing resulted in an average of 29.5 seeds per flower, as compared to 59.9 seeds per flower from hand-pollination of stigmas with self pollen. In an assessment of the possible significance of this selfing mode, emasculated flowers did not set significantly fewer seeds in a natural stand in Ohio, suggesting that few seeds resulted from autonomous selfing at that site. Modest inbreeding depression was detected at this population. Our results suggest that delayed autonomous selfing is more common in northern populations, where it may facilitate population establishment and persistence at times when pollinators are scarce.     

Comparative pollination biology in two sympatric varieties of Cypripedium macranthos (Orchidaceae) on Rebun Island,Hokkaido, Japan

Cypripedium macranthos sensu lato typically has purple‐pink flowers with no nectar and harvestable pollen. On Rebun Island, Hokkaido, Japan, purple‐pink‐flowered C. macranthos var. macranthos individuals rarely grow among numerous pale‐cream‐flowered C. macranthos var. rebunense plants. In both varieties, flower size is similar, their flowering periods completely overlap, and they share the same pollinator (Bombus pseudobaicalensis). However, in only one of 12 years from 2001 to 2012 did var. macranthos attain an annual fruit‐set ratio (an estimate of pollination success) higher than that of sympatric var. rebunense plants. These findings strongly suggest that in C. macranthos on Rebun Island, flower color results in the differential pollination success, because the pollinator prefers pale‐cream Cypripedium flowers and/or avoids purple‐pink ones, thereby producing pollinator‐mediated selection favoring pale‐cream flowers.     

Gynostemium micromorphology and pollination in Epipactis microphylla (Orchidaceae)

Self-pollination conditions have been recorded frequently in local populations of the genus Epipactis, and structural modifications have sometimes been reported, supporting taxonomic recognition of new species. As part of a survey of gynostemium micromorphology of Italian Epipactis populations, we studied an Apennine population of Epipactis microphylla (Ehrh.) Sw. a species listed as autogamous but with residual allogamous characters. In this population we observed the tendency to perform pre-anthesis cleistogamy. However, self-pollination was not a consequence of modification in column morphology. Cryptic pollen germination occurred in anther thecae or in the clinandrium, bypassing any contact with the receptive stigma. Pollen germination started from the distal pool of the pollinium, close to the rostellum-viscidium, suggesting that the rostellum-viscidium may have a function in promoting pollen-tube growth. Germinated pollen was exposed at the anthesis.     

Reproductive biology and pollination ecology of Triplaris gardneriana (Polygonaceae): a case of ambophily in the Brazilian Chaco

• Triplaris gardneriana (Polygonaceae) is a dioecious pioneer tree reported as insect‐pollinated, despite possessing traits related to anemophily. Here, we analyse the possible roles of insects and wind on the pollination of this species to establish whether the species is ambophilous.
• We carried out observations of floral biology, as well as on the frequency and behaviour of pollinators visiting flowers in a population of T. gardneriana in the Chaco vegetation of Brazil. We conducted experimental pollinations to determine the maternal fertility of female plants and whether they were pollen‐limited, and we also conducted aerobiological experiments to provide evidence of how environmental factors influence atmospheric pollen dispersal.
• The population comprised an area of approximately 152.000 m² and was composed of 603 female and 426 male plants (sex ratio = 0.59:0.41). We observed 48 species of insects visiting flowers of T. gardneriana, of which the bees Scaptotrigona depilis and Apis mellifera scutellata were the most effective pollinators. We recorded pollen grains dispersed by wind on 74% of the glass slides placed on females, located at different distances (1–10 m) from male plants.
• Airborne pollen concentration was negatively correlated with relative humidity and positively correlated with temperature. Our observations and experimental results provide the first evidence that T. gardneriana is an ambophilous species, with pollen dispersal resulting from both animal and wind pollination. This mixed pollination strategy may be adaptive in T. gardneriana providing reproductive assurance during colonisation of sites with different biotic and abiotic conditions.

     

Dioecy and wind pollination in Pernettya rigida (Ericaceae) of the Juan Fernández Islands

Pernetttya rigida is endemic to the Juan Fernández archipelago. Although all flowers are complete, with seemingly fertile stamens and pistils, differences in fruit production and detailed field, anatomical, and morphological studies indicate they are functionally unisexual, and the species is consequently dioecious. A comparison of 15 features demonstrated sigdicant differences between the sexes. The populational sex ratio is 1:1. Nectaries located between the filaments produce small amounts of floral nectar with similar sugar composition in both sexes. There are =33,357 (or = 133,429) pollen tetrads (or pollen grains)/male flower and =109 ovules/female flower. No tetrads in either hand self-pollinated or open pollinated male flowers showed any pollen germination. Tetrads on open- and hand-pollinated female flowers germinate. Female flowers do not show autogamy or apomixis. During more than 80 hours of field observation, we recorded only seven floral visitors (representing three insect species). In spite of this, openpollinated female flowers have abundant fruit and seed-set. Thus, we conclude that pollen is transferred abiotically and the ever-present wind over the exposed ridges of the islands is the likely dispersal agent. A number of anemophilous features, such as dry pollen and exposed habitat, support this conclusion. Thus, dioecy and anemophily have evolved independently, in situ , in this species in this remote locality. Preservation of habitat and elimination of competitive invasives are the primary conservation challenges.     

Floral biology of two Vanilloideae (Orchidaceae) primarily adapted to pollination by euglossine bees

Vanilloideae comprises 15 genera distributed worldwide, among which are Vanilla and Epistephium (tribe Vanilleae). Based on field and laboratory investigations, the pollination biology of V. dubia and E. sclerophyllum was analysed. The former was surveyed in a semi‐deciduous mesophytic forest at the biological reserve of Serra do Japi and in a marshy forest at the city of Pradópolis, southeastern Brazil. The latter was examined in rocky outcrop vegetation in the Chapada Diamantina, northeastern Brazil. In the studied populations, the tubular flowers of V. dubia and E. sclerophyllum were pollinated by bees. Pollen was deposited on either their scutellum (V. dubia) or scutum (E. sclerophyllum). The mentum region of V. dubia is dry, whereas that of E. sclerophyllum presents a small quantity of dilute nectar. Flowers of E. sclerophyllum are scentless, while those of V. dubia are odoriferous. Although V. dubia is self‐compatible, it needs a pollinator to produce fruit. In contrast, E. sclerophyllum sets fruit through spontaneous self‐pollination, but biotic pollination also occurs. Both species are primarily adapted to pollination by euglossine bees. Pollination by Euglossina seems to have occurred at least twice during the evolution of Vanilleae. Furthermore, shifts between rewarding and reward‐free flowers and between autogamous and allogamous species have been reported among vanillas.     

Spatio‐temporal patterns in pollination of deceptive Aristolochia rotunda L. (Aristolochiaceae)

• Pollination success of highly specialised flowers is susceptible to fluctuations of the pollinator fauna. Mediterranean Aristolochia rotunda has deceptive trap flowers exhibiting a highly specialised pollination system. The sole pollinators are kleptoparasitic flies in search of food. This study investigates these pollinators on a spatio‐temporal scale and the impact of weather conditions on their availability. Two potential strategies of the plants to cope with pollinator limitation, i.e. autonomous selfing and an increased floral life span, were tested.
• A total of 6156 flowers were investigated for entrapped pollinators in 10 Croatian populations. Availability of the main pollinator was correlated to meteorological data. Artificial pollination experiments were conducted and the floral life span was recorded in two populations according to pollinator availability.
• Trachysiphonella ruficeps (Chloropidae) was identified as dominant pollinator, along with less abundant species of Chloropidae, Ceratopogonidae and Milichiidae. Pollinator compositions varied among populations. Weather conditions 15–30 days before pollination had a significant effect on availability of the main pollinator. Flowers were not autonomously selfing, and the floral life span exhibited considerable plasticity depending on pollinator availability.
• A. rotunda flowers rely on insect pollen vectors. Plants are specialised on a guild of kleptoparasitic flies, rather than on a single species. Pollinator variability may result in differing selection pressures among populations. The availability/abundance of pollinators depends on weather conditions during their larval development. Flowers show a prolonged trapping flower stage that likely increases outcrossing success during periods of pollinator limitation.

     

Factors affecting germinable seed yield in Cypripedium calceolus var. pubescens (Willd.) Correll and Epipactis helleborine (L.) Crantz (Orchidaceae)

One of the challenges facing orchid conservationists is to get a better understanding of seed parent fecundity. This can be especially laborious for orchids whose seed is difficult to germinate. Mature seeds ofCypripedium calceolusvar.pubescens(Willd.) Correll (Orchidaceae) have been reported to require 8 weeks pre-chilling at 4°C and a further month at 20°C to induce germination, but seeds prematurely harvested at 7–8 weeks post-pollination germinate asymbiotically without chilling in less than 1 month. Mature seeds ofEpipactis helleborine(L.) Crantz (Orchidaceae) have also presented a challenge but the identification of plants whose seeds are quick and easy to germinate has provided us with a tool to study patterns of production of this seed class. Preliminary field experiments suggest that timing of pollination in these two taxa is critical for the development of seed. When seeds from capsules ofC. calceolusvar.pubescensharvested 7 weeks after pollination were examined, the proportion of fully mature embryos increased with increasing anthesis-pollination interval over the first 10 days. Paired flowers from five genets xenogamously hand-pollinated at 2-day intervals from the second day after anthesis until the flowers wilted were significantly less likely to produce fruits as the flowers aged but the flower age at which this occurred varied between genets. The germinability of seeds from different pollen and seed parent combinations was tested on eight different media formulations and over a range of post-anthesis and post-pollination intervals. Different aged seeds germinated variably, the post-anthesis interval being the most reliable predictor of germination outcomes. Intercrossing of plants ofC. calceolusvar.pubescenssuggests that seed production and germinability are maternally mediated traits. WithE. helleborine, germinable seed can be produced both by self- and by cross-pollination of selected plants but we have observed an influence of flower position in the inflorescence on self-compatibility and on the production of the easy-to-germinate seed class. The timing of pollination in these two unrelated orchids has a considerable influence upon seed production and seed quality in plants of established fecundity. These findings suggest that flower age at pollination must be considered when interpreting similar studies in the Orchidaceae.     

Recurrent polymorphic mating type variation in Madagascan Bulbophyllum species (Orchidaceae) exemplifies a high incidence of auto‐pollination in tropical orchids

The transition from outcrossing to self‐fertilization is one of the most common evolutionary changes in angiosperms. The orchid family exemplifies this evolutionary trend but, because of a general lack of large‐scale surveys on auto‐pollination in orchid taxa, the incidence and modes of auto‐pollination among (sub)tropical orchids remain poorly known. In the present study, we assessed the frequency and mode of auto‐pollination within and among species of a largely monophyletic group of Madagascan Bulbophyllum. The capacity for autonomous fruit set was investigated by bagging experiments in the greenhouse and the field, complemented with detailed floral micromorphological studies of the gynostemium. Our survey comprises 393 accessions, representing at least 78 species, and thus approximately 37% of the species diversity of the genus in the Madagascan region. Our studies revealed that mating type is directly related to gynostemium structure, most often involving the presence or absence of a physical barrier termed ‘rostellum’. As a novel and unexpected finding, we identified eight species of a single lineage of Madagascan Bulbophyllum (termed ‘clade C’), in which auto‐pollinating morphs (selfers), either lacking a rostellum or (rarely) possessing a stigmatic rostellum, co‐exist with their pollinator‐dependent conspecifics (outcrossers). We hypothesize that auto‐pollination via rostellum abortion has a simple genetic basis, and probably evolved rapidly and recurrently by subtle changes in the timing of rostellum development (heterochrony). Thus, species of clade C may have an intrinsic genetic and developmental lability toward auto‐pollination, allowing rapid evolutionary response under environmental, perhaps human‐disturbed conditions favouring reproductive assurance. Overall, these findings should stimulate further research on the incidence, evolution, and maintenance of mating type variation in tropical orchids, as well as how they adapt(ed) to changing environmental conditions. © 2014 The Authors. Botanical Journal of the Linnean Society published by John Wiley & Sons Ltd on behalf of The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 242–258.     

The pollination of Habenaria rhodocheila (Orchidaceae) in South China: When butterflies take sides

Habenaria is one of the largest terrestrial genera in the family Orchidaceae. Most field studies on Habenaria species with greenish–white and nocturnal scented flowers are pollinated by nocturnal hawkmoths and settling moths. However, H. rhodocheila presents reddish flowers lacking a detectable scent and fails to fit the moth pollination syndrome. We investigated the pollinators, breeding system, and functional traits of H. rhodocheila in South China and found that two diurnal swallowtail butterflies Papilio helenus and Papilio nephelus (Papilionidae) were the effective pollinators. When butterflies foraged for nectar in the spur, the pollinia became attached between the palpi. A triangular projected median rostellar lobe was found at the entrance (sinus) of the spur of H. rhodocheila. This lobe divided the spur opening into two entrances forcing butterflies to enter their proboscides through the left or right side. When the projection of median rostellar lobe was removed, the site of pollinium attachment changed to the eyes of the butterflies, leading to a higher rate of pollinium removal but lower rate of pollinium deposition. Our quartz glass cylinder choice experiment suggested that visual rather than olfactory cues provided the major stimuli for butterflies to locate these flowers. Hand pollination experiments suggested this species was self‐compatible but pollinator‐dependent. However, the proportion of seeds with large embryos produced in self‐pollinated fruits was significantly lower than in cross‐pollinated fruits, indicating a significant inbreeding depression. Unlike many other orchid species, fruit set was higher than rates of pollinium removal, indicating a high level of pollination efficiency in a species with friable pollinia. Shifts from moth to butterfly pollination in the genus Habenaria parallel other orchid lineages providing insights into the potential for pollinator‐mediated floral trait selection.     

At daybreak--reproductive biology and isolating mechanisms of Cirrhaea dependens (Orchidaceae)

Floral biology, reproduction, pollinator specificity, and fruit set of Cirrhaea dependens were recorded in forest areas of Southeastern Brazil. Cirrhaea dependens is a lithophytic or epiphytic herb occurring very sparsely below dense canopies. Nearly all the flowers of a single plant open simultaneously before dawn. They are short-lasting and offer floral fragrances as rewards, which are collected by male euglossine bees. Observations carried out in mesophytic forests at Serra do Japi revealed that Euglossa VIRIDIS is their principal pollinator, with Eufriesea violacea a sporadic co-pollinator. Visitation started soon after flower opening, and attractiveness remained high for about 2 h, decreasing abruptly at sunrise. Flower anthesis with subsequent fragrance release seems to be correlated with bee attraction. Observations using chemical baits were carried out at Serra do Japi, and in the Atlantic Rain Forest of Picinguaba. Three euglossine species were captured with pollinaria of C. dependens in Picinguaba, whereas only E. violacea was attracted in Serra do Japi. In Picinguaba, C. dependens occurs sympatrically with C. saccata and C. loddigesii. Each species attracted different pollinators. The specificity and resulting reproductive isolation are due to the production of different odours by each orchid species. Cirrhaea dependens is self-compatible but pollinator-dependent. The reproductive success was low and appears to result from a combination of factors discussed here, such as the production of short-lived flowers, presence of floral mechanisms avoiding self-pollination, non-synchronization of flower phases among plants, and populations with few and sparsely distributed individuals.     

Comparative study of pollination biology of two closely related alpine Primula species,namely Primula beesiana and P. bulleyana (Primulaceae)

Abstract The comparative pollination biology of a population of Primula beesiana, a population of P. bulleyana, and an overlapping population of these two species in Yulong Shan, Yunnan Province, China, was studied in 2004 and 2005. The results indicate that both P. beesiana and P. bulleyana are typical heterostylous and obligate outcrossing species; the main pollinators of the two species were bees and butterflies. At the sites of all three populations, the main pollinating visitors of the two species showed preference for one of the two species, resulting in the ethological isolation of the two species by the pollinators. This ethological isolation contributes to the reproductive isolation of the two species, which supports the hypothesis that P. beesiana and P. bulleyana are two distinct species. The reproductive isolation of the two species (ethological isolation) is probably an important mechanism in maintaining species boundaries in the genus and has contributed to the species diversification of Primula in the area. In addition, gene exchanges between P. beesiana and P. bulleyana has occurred to some extent in the overlapping population, but whether natural hybridization has contributed to species diversification in Primula remains to be determined.     

Comparative study of pollination biology of two closely related alpine Primula species,namely Primula beesiana and P.bulleyana (Primulaceae)

The comparative pollination biology of a population of Primula beesiana,a population of P.bulleyana,and an overlapping population of these two species in Yulong Shan,Yunnan Province,China,was studied in 2004 and 2005.The results indicate that both P.beesiana and P.bulleyana are typical heterostylous and obligate outcrossing species;the main pollinators of the two species were bees and butterflies.At the sites of all three populations,the main pollinating visitors of the two species showed preference for one...     

Breeding system and thrips (Thysanoptera) pollination in the endangered tree Ocotea porosa (Lauraceae): implications for conservation

Ocotea porosa has been extensively exploited over the past few decades because of the quality of its wood. Today, populations are reduced and the species is now included in the Red List of threatened species by the International Union for Nature Conservation. For conservation and management purposes, it is extremely important that we understand its reproductive ecology. Floral morphology was described based on field and scanning electron microscopic examination. The reproductive system was determined through experimentally controlled pollination along with observations of pollen tube growth. Pollinators were identified through field observations. Flowers of O. porosa are small, shallow, inconspicuous, asynchronous and grouped in inflorescences. This species presented self‐compatibility, but did not reproduce through apomixy and spontaneous self‐pollination was very rare (5%). Despite being monoclinal, flowers were protogynic, and the gynoecium was receptive after the first day of anthesis when the anthers were closed. Spontaneous self‐pollination was avoided by the extrorse position of the anthers of the internal stamens. Frankliniella gardeniae (Thysanoptera) was the only pollinating species observed and, after visiting several asynchronous flowers in the same inflorescence, favors geitonogamy. Together the small distance supposedly achieved by thrips in flight and the small population density of O. porosa can reduce the chances of cross‐pollination in this species. If so, conservation measures must include preservation of the current population and possibly planting to increase population density. This would reduce the distance between individuals and increase genetic variability. Thrips as pollinators must be included in conservation planning for O. porosa.