Comparison of floral properties and breeding system in dimorphic Buddleja delavayi (Scrophulariaceae)

In this study, floral color, scent composition and emission rate, nectar property, pollinators, and breeding system of dimorphic Buddleja delavayi Gagnep. were investigated. Flower color of B. delavayi was determined using a standard color chart and spectrophotometer, and two distinct color polymorphisms were observed having purple or white flowers. Floral scents of B. delavayi were collected using dynamic headspace adsorption and identified with coupled gas chromatography and mass spectrometry. In total, 28 compounds were identified from the flowers of B. delavayi. The identified scents were divided into three chemical classes based on their biosynthetic origin: terpenes, fatty acid derivatives, and benzenoids. The scent profiles in all individuals were dominated by a few components, such as lilac aldehyde and alcohol, 4-oxoisophorone, benaldehyde, and oxoisophorone oxide. Floral scent composition (benzenoids and terpenes) showed a significant difference between white and purple flower morphs. Flower color–flower scent associations in B. delavayi were identified with two distinct scent profiles in the two color phenotypes. The studies of other floral characteristics (nectar, floral visitors, breeding system, and fruit set) indicated that floral scent emission rate, nectar volume, visitor visitation frequency, and natural fruit set were not significantly different between the two flower color morphs. Bagging experiments revealed that seed production of B. delavayi is dependent mainly on honeybee Apis cerana. Lastly, this study implies that dimorphic floral color in B. delavayi may have been maintained by floral visitors and nectar guide color.     

Nocturnal emission and post-pollination change of floral scent in the leafflower tree,Glochidion rubrum,exclusively pollinated by seed-parasitic leafflower moths

Many insect-pollinated plants use floral scent signals to attract and guide the effective pollinators, and temporal patterns of their floral scent emission may be tuned to respond to the pollinator's activity and pollination status. In the intimate nursery pollination mutualism between monoecious Glochidion trees (Phyllanthaceae) and Epicephala moths (Gracillariidae), floral scent signals mediate species-specific interactions and influence the moth's efficient pollen-collecting and pollen-depositing behaviors on male and female flowers, respectively. We tested the hypotheses that both sexes of flowers of Epicephala-pollinated Glochidion rubrum exhibit a diel pattern of scent emission matching the activity period of the nocturnally active pollinator, and that female flowers change the chemical signal after pollination to reduce further visits and oviposition by the pollinator. We investigated the diel change of floral scent emissions during two consecutive days and the influence of pollination on the floral scent by conducting hand-pollinations in the field. The total scent emission of male and female flowers was higher at night than in the day, which would be expected from the nocturnal visitations of Epicephala moths. Some compounds exhibited a clear nocturnal emission rhythm. Hand-pollination experiments revealed that emission of two compounds, nerolidol and eugenol, drastically decreased in pollinated flowers, suggesting that these compounds may function as key attractants for the pollinator; however, the total scent emission of the female flower was not influenced by hand-pollination. The pattern of the floral scent emission of G. rubrum may be optimized to attract nocturnal pollinators and reduce oviposition.     

花颜色和花气味的量化研究方法

花颜色和花气味是花部构成的重要内容。在已开展的传粉生态学研究中对二者的报道主要是描述性的,而其量化研究可以为揭示传粉机制提供有力的实验证据。本文主要介绍了花颜色的测量和标定方法,包括比色卡、分光色差仪和便携式光谱仪等;花气味的采集方法,包括动态顶空套袋-吸附采集法、吸附-溶剂洗脱法和固相微萃取法等;花气味的检测和分析方法,包括气相色谱-质谱联用仪分析和电子鼻型超速气相色谱仪分析等;以及昆虫行为学实验方法,包括气相色谱-昆虫触角电位联用技术、Y型嗅觉仪和飞行箱实验等。科研人员可以根据实验材料的特点和实验目的选择适合的量化研究方法。     

Natural selection on floral volatile production in Penstemon digitalis: Highlighting the role of linalool

Natural selection is thought to have shaped the evolution of floral scent; however, unlike other floral characters, we have a rudimentary knowledge of how phenotypic selection acts on scent. We found that floral scent was under stronger selection than corolla traits such as flower size and flower color in weakly scented Penstemon digitalis. Our results suggest that to understand evolution in floral phenotypes, including scent in floral selection, studies are crucial. For P. digitalis, linalool was the direct target of selection in the scent bouquet. Therefore, we determined the enantiomeric configuration of linalool because interacting insects may perceive the enantiomers differentially. We found that P. digitalis produces only (S)-(+)-linalool and, more interestingly, it is also taken up into the nectar. Because the nectar is scented and flavored with (S)-(+)-linalool, it may be an important cue for pollinators visiting P. digitalis flowers.     

Odour and colour polymorphism in the food-deceptive orchid Dactylorhiza romana

The food deceptive orchid, Dactylorhiza romana (Sebastiani) Soó exhibits a colour polymorphism with yellow, red, and intermediate orange morphs. In this study we tested if floral odour differed among the three distinct colour morphs. We identified 23 odour compounds in D. romana, and all of them occurred in the three colour morphs. Monoterpenes dominated the floral scent. On the basis of Euclidean distances of relative amounts of compounds, yellow morphs were closer to each other than to orange or red morphs. Differentiation of the morphs was mainly due to linalool and benzaldehyde. Linalool occurred in low relative amounts in the yellow morphs, but in high amounts in some of the red individuals, whereas benzaldehyde occurred in higher relative amounts in yellow morphs. Linalool and benzaldehyde are known to be important signal-substances in plant-insect communication, however, it remains to be shown whether insects can discriminate between flower morphs on the basis of the here shown odour differences.     

Comparison of floral traits and pollinator assemblages of insular and mainland varieties of Lilium auratum

Lilium auratum var. platyphyllum, which is endemic to the Izu Islands, has white petals and emits a strong floral scent, typical of moth‐pollinated plants. Moths might be important pollinators within the Izu Islands because swallowtail butterflies are absent or rare there. Therefore, we investigated insular var. platyphyllum and widespread L. auratum var. auratum on the mainland of Japan to clarify the relationship between floral characteristics and pollinators. Measurement of floral scent intensity using an odor sensor indicated that the intensity increased in the evening and at night in var. platyphyllum, whereas intensity increased at night in var. auratum. Total sugar weight in nectar, which was calculated by nectar volume and sugar concentration, showed that the flowers of var. platyphyllum secreted nectar abundantly both in the evening and at night, whereas those of var. auratum secreted an almost constant amount of nectar throughout the day. Flower visitor assemblages and their frequencies studied using digital cameras suggested that crepuscular and nocturnal hawkmoths are important pollinators for var. platyphyllum. In contrast, both diurnal swallowtail butterflies and nocturnal large hawkmoths are important and effective pollinators for var. auratum. These conclusions were also supported by the exclusion experiments of either diurnal or nocturnal flower visitors using mesh bags. The rates of seed sets of var. platyphyllum were significantly higher from nocturnal pollination than from diurnal pollination. Thus, this study revealed that floral traits of var. platyphyllum show more adaptation for crepuscular and nocturnal hawkmoths, which are relatively abundant in the Izu Islands, than those of var. auratum.     

Temporal variation of flower scent in Silene otites (Caryophyllaceae): a species with a mixed pollination system

Floral scent composition and emission are often related to olfactory abilities, preferences and activity times of pollinating agents. However, most studies have focused on species with one pollinator type, and little is known about species with mixed pollination systems. We investigated the temporal variation in flower visitor activity and flower scent emission of Silene otites (Caryophyllaceae), a species regarded as being nocturnal and pollinated by moths and mosquitoes. We found, however, that S. otites is not only visited at night by moths and mosquitoes, but also by a wide range of day‐active insects. Furthermore, both the intensity and composition of floral scent change during the course of the day. Total scent emission reached its maximum in the early hours of the night when compounds known to be attractants for moths and/or mosquitoes, such as lilac aldehyde, benzaldehyde, phenylacetaldehyde and 2‐phenylethanol, dominated the scent. However, although less intensive, emission continued during the day. In the early morning, α‐pinene, hotrienol, 2‐phenylethanol and δ‐3‐carene were the dominant scent compounds. (E)‐β‐Ocimene, hotrienol and benzyl alcohol contributed proportions of at least 10% before midday when flies, such as syrphids, were abundant flower visitors, whereas linalool and hotrienol contributed to the scent with at least 10% each in the afternoon when bees also contributed to flower visits. We conclude that, in S. otites, the changing emission patterns are ‘in tune’ with the olfactory abilities, preferences and activity times of its day‐ and night‐active potentially pollinating flower visitors. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169 , 447–460.     

Spatial and temporal patterns of floral scent emission in Dianthus inoxianus and electroantennographic responses of its hawkmoth pollinator

Scent emission is important in nocturnal pollination systems, and plant species pollinated by nocturnal insects often present characteristic odor compositions and temporal patterns of emission. We investigated the temporal (day/night; flower lifetime) and spatial (different flower parts, nectar) pattern of flower scent emission in nocturnally pollinated Dianthusinoxianus, and determined which compounds elicit physiological responses on the antennae of the sphingid pollinator Hyles livornica.The scent of D.inoxianus comprises 68 volatile compounds, but is dominated by aliphatic 2-ketones and sesquiterpenoids, which altogether make up 82% of collected volatiles. Several major and minor compounds elicit electrophysiological responses in the antennae of H. livornica. Total odor emission does not vary along day and night hours, and neither does along the life of the flower. However, the proportion of compounds eliciting physiological responses varies between day and night. All flower parts as well as nectar release volatiles. The scent of isolated flower parts is dominated by fatty acid derivatives, whereas nectar is dominated by benzenoids. Dissection (= damage) of flowers induced a ca. 20-fold increase in the rate of emission of EAD-active volatiles, especially aliphatic 2-ketones.We suggest that aliphatic 2-ketones might contribute to pollinator attraction in D. inoxianus, even though they have been attributed an insect repellent function in other plant species. We also hypothesize that the benzenoids in nectar may act as an honest signal (‘nectar guide’) for pollinators.     

The evolution of floral scent: the influence of olfactory learning by insect pollinators on the honest signalling of floral rewards

1.  The evolution of flowering plants has undoubtedly been influenced by a pollinator's ability to learn to associate floral signals with food. Here, we address the question of 'why' flowers produce scent by examining the ways in which olfactory learning by insect pollinators could influence how floral scent emission evolves in plant populations.
2.  Being provided with a floral scent signal allows pollinators to learn to be specific in their foraging habits, which could, in turn, produce a selective advantage for plants if sexual reproduction is limited by the income of compatible gametes. Learning studies with honeybees predict that pollinator-mediated selection for floral scent production should favour signals which are distinctive and exhibit low variation within species because these signals are learned faster. Social bees quickly learn to associate scent with the presence of nectar, and their ability to do this is generally faster and more reliable than their ability to learn visual cues.
3.  Pollinators rely on floral scent as a means of distinguishing honestly signalling flowers from deceptive ones. Furthermore, a pollinator's sensitivity to differences in nectar rewards can bias the way that it responds to floral scent. This mechanism may select for flowers that provide olfactory signals as an honest indicator of the presence of nectar or which select against the production of a detectable scent signal when no nectar is present.
4.  We expect that an important yet commonly overlooked function of floral scent is an improvement in short-term pollinator specificity which provides an advantage to both pollinator and plant over the use of a visual signal alone. This, in turn, impacts the evolution of plant mating systems via its influence on the species-specific patterns of floral visitation by pollinators.     

报春石斛花香气成分及其日变化规律

采用静态顶空气相色谱-质谱(GC-MS)联用技术研究报春石斛(Dendrobium primulinum)花香气成分及香气释放的日变化规律,检测并分析一天内六个时间点(6:00、9:00、12:00、15:00、18:00、21:00)报春石斛花的香气成分。各时间点占总香气释放量排序为：6时(35.26%)>9时(22.78%)>12时(19.35%)>21时(7.86%)>15时(7.59%)>18时(7.17%)。共检测出202种香气物质,包括酯类、烯萜类、醇类、醛类、烷类等,其中6:00有93种,9:00有87种,12:00有68种,15:00有61种,18:00有63种,21:00有65种。各个时间点共有的化合物21种。报春石斛花香气释放量6:00最高,18:00最低,主要香气成分是醋酸辛酯。     

Priority of color over scent during flower visitation by adult <Emphasis Type="Italic">Vanessa indica</Emphasis> butterflies

Most flower visitors innately prefer a particular color and scent, and use them as cues for flower recognition and selection. However, in most cases, since color and scent serve as a combined signal, not only does the preference for an individual cue, but also the preference hierarchy among different cues, influence their flower visitation. In the present study, we attempted to reveal (1) the chromatic and (2) the olfactory cues that stimulate flower visiting, and (3) the preference hierarchy between these cues, using the naïve adult butterfly Vanessa indica. When we offered 12 different-colored (six chromatic and six achromatic) paper flower models, V. indica showed a color preference for yellow and blue. When we examined the proboscis extension reflex (PER) of V. indica towards 16 individual compounds identified in the floral scents from two nectar plants belonging to the family Compositae, Taraxacum officinale and Cirsium japonicum, six compounds were found to have relatively high PER-eliciting activities, including benzaldehyde, acetophenone, and (E+Z)-nerolidol. When we combined color and scent cues in two-choice bioassays, where butterflies were offered flower models that were purple (a relatively unattractive color), the models scented with these active compounds were significantly more attractive than the odorless controls. In addition, synthetic blends mimicking the floral scents of T. officinale and C. japonicum (at doses equivalent to that of ten flowers) enhanced the number of visits to the scented models. However, the effect of odorizing was not conspicuous in parallel bioassays when yellow flower models were used, and the butterflies also significantly preferred odorless yellow models to scented purple models. These results demonstrate that V. indica depends primarily on color and secondarily on scent during flower visitation.     

Is the timing of scent emission correlated with insect visitor activity and pollination in long‐spurred Satyrium species?

Plants are expected to emit floral scent when their pollinators are most active. In the case of long‐tubed flowers specialised for pollination by crepuscular or nocturnal moths, scent emissions would be expected to peak during dawn. Although this classic idea has existed for decades, it has rarely been tested quantitatively. We investigated the timing of flower visitation, pollination and floral scent emissions in six long‐spurred Satyrium species (Orchidaceae). We observed multiple evening visits by pollinaria‐bearing moths on flowers of all study species, but rarely any diurnal visits. The assemblages of moth pollinators differed among Satyrium species, even those that co‐flowered, and the lengths of moth tongues and floral nectar spurs were strongly correlated, suggesting that the available moth pollinator fauna is partitioned by floral traits. Pollinarium removal occurred more frequently during the night than during the day in four of the six species. Scent emission, however, was only significantly higher at dusk than midday in two species. Analysis of floral volatiles using gas chromatography coupled with mass spectrometry yielded 168 scent compounds, of which 112 were species‐specific. The scent blends emitted by each species occupy discrete clusters in two‐dimensional phenotype space, based on multivariate analysis. We conclude that these long‐spurred Satyrium species are ecologically specialised for moth pollination, yet the timing of their scent emission is not closely correlated with moth pollination activity. Scent composition was also more variable than expected from a group of closely related plants sharing the same pollinator functional group. These findings reveal a need for greater understanding of mechanisms of scent production and their constraints, as well as the underlying reasons for divergent scent chemistry among closely related plants.     

Variation in scent emission among floral parts and inflorescence developmental stages in beetle-pollinated Protea species (Proteaceae)

Floral fragrances are an important component for pollinator attraction in beetle-pollinated flowers. Several genera in the Proteaceae contain beetle-pollinated species. However, there is no information on the floral scent chemistry of beetle-pollinated members of the family. In this paper we report on the spatial variation and differences between developmental stages in emission of inflorescence (flowerhead) volatiles of four South African Protea species (P. caffra, P. dracomontana, P. simplex, and P. welwitschii) that are pollinated by cetoniine beetles. The scents from different inflorescence parts (bracts, perianth, styles, and nectar) and from successive anthesis stages of whole inflorescences were sampled using dynamic headspace collection and identified using GC–MS. Although the four species shared many scent compounds, possibly reflecting their close phylogenetic relationships and common pollinators, they showed significant differences in overall scent composition due to various species-specific compounds, such as the unique tiglate esters found in the scent of P. welwitschii. The strongest emissions and largest number of volatiles, especially monoterpenes, were from inflorescences at full pollen dehiscence. Senescing inflorescences of two species and nectars of all species emitted proportionally high amounts of acetoin (3-hydroxy-2-butanone) and aromatic alcohols, typical fermentation products. As a consequence, the scent composition of nectar was much more similar among species than was the scent composition of other parts of the inflorescence. These results illustrate how the blends of compounds that make up the overall floral scent are a dynamic consequence of emissions from various plant parts.     

Reward quality influences the development of learned olfactory biases in honeybees

Plants produce flowers with complex visual and olfactory signals, but we know relatively little about the way that signals such as floral scents have evolved. One important factor that may direct the evolution of floral signals is a pollinator''s ability to learn. When animals learn to associate two similar signals with different outcomes, biases in their responses to new signals can be formed. Here, we investigated whether or not pollinators develop learned biases towards floral scents that depend on nectar reward quality by training restrained honeybees to learn to associate two similar odour signals with different outcomes using a classical conditioning assay. Honeybees developed learned biases towards odours as a result of differential conditioning, and the extent to which an olfactory bias could be produced depended upon the difference in the quality of the nectar rewards experienced during conditioning. Our results suggest that differences in reward quality offered by flowers influence odour recognition by pollinators, which in turn could influence the evolution of floral scents in natural populations of co-flowering plants.     

Inflorescence scent,color, and nectar properties of “butterfly bush” (Buddleja davidii) in its native range

In this study, flower color, nectar properties, and inflorescence scent composition of eight natural and one introduced Buddleja davidii populations were investigated. Flower color of B. davidii was determined using the Royal Horticultural Society Color Chart and ranged from purple to white. Volume of nectar produced by a single flower ranged from 0.36 μl to 0.64 μl and total sugar concentration produced by inflorescence ranged from 17.0% to 33.5% in all populations. Floral nectar volume and sugar concentration were not significantly different between two flower color morphs in the B. davidii populations. Floral scents of B. davidii were collected using dynamic headspace adsorption and identified with coupled gas chromatography and mass spectrometry. In total, 33 compounds were identified from the inflorescences of B. davidii. The identified scents were divided into five chemical classes based on their biosynthetic origin: irregular terpenes, monoterpenoids, sesquiterpenoids, fatty acid derivatives, and benzenoids. The scent profiles in all populations were dominated by few components, such as: 4-oxoisophorone, E,E-α-farnesene, and 1-octen-3-ol. Given that inflorescence scents from natural and introduced individuals coming from the same population have discrepant chemical composition, we infer that phenotype plasticity may mediate floral scent composition. Based on the comparison of present and other data available on floral scent in B. davidii, we conclude that inflorescence scent may serve as a specific signal helping to attract pollinating butterflies to locate flowers as nectar sources, and may have evolved in conjunction with the sensory capabilities of butterflies and moths as a specific group of pollinators.     

Pollinator attraction in Cornus capitata (Cornaceae): the relative role of visual and olfactory cues

Aims It is generally accepted that visual displays and floral scent play important roles in communication between flowering plants and their pollinators. However, the relative role of visual and olfactory cues in pollinator attraction is largely unknown. In this study, we determined the roles of both types of cue in attracting pollinators to Cornus capitata, a medium sized tree with each capitulum surrounded by four large, white, petaloid bracts.Methods Pollinator observations and pollination experiments were conducted in a natural population; the inflorescences' visual and olfactory signals were characterized by spectral and chemical analyses; the responses of pollinators to visual and olfactory cues were tested using dual-choice behavioural bioassays; the relative roles of visual and olfactory cues in pollinator attraction were tested by comparing the responses of pollinators to inflorescences subjected to three experimental treatments (intact, all bracts removed, and capitulum removed) within the natural population.Important findings For fruit set, C. capitata is entirely dependent on pollinators, with a bee, Anthophora sp., being the main pollinator. Bracts present high colour distance and green contrast against the leaves. Twelve volatile compounds in the floral scent were detected, most of which have previously been reported to be attractive to a broad spectrum of bee species. Behavioural bioassays showed that both, visual cues alone and olfactory cues alone, are attractive to pollinating bees. However, visual cues alone attracted significantly more approaches than olfactory cues alone, while olfactory cues alone elicited a significantly higher landing percentage than visual cues alone. The finding suggests that, in the C. capitata – Anthophora sp. interaction, visual cues are mainly used for location from long distances, while olfactory cues mainly aid landing from short distances. Our results indicate that different modalities of floral cues should be considered together to understand fully the communication between flowering plant and pollinators.     

Diel fragrance pattern correlates with olfactory preferences of diurnal and nocturnal flower visitors in Salix caprea (Salicaceae)

Floral scent, often a complex mixture of several volatile organic compounds (VOCs), has generally been interpreted as an adaptation to attract pollinators. However, not many studies have analysed which VOCs are functionally relevant for the reproductive success of a plant. Here, we show that, in Salix caprea (Salicaceae), temporal changes in floral scent emission during the day and night attract two different types of flower visitor: bees during the day and moths during the evening and night. We analysed the contribution of the two flower visitor groups to the reproductive success of the plant. The differences in scent emitted during the peak activity times of flower visitors (day versus night) were quantified and the response of 13 diurnal/nocturnal pollinator taxa to the floral scents was tested using gas chromatographic and electroantennographic techniques. Many of the c. 40 identified scent compounds were physiologically active, and bees and moths responded to nearly identical sets of compounds, although the response strengths differed. In bioassays, bees preferred the most abundant 1,4‐dimethoxybenzene over lilac aldehyde, a compound with increased emission at night, whereas moths preferred lilac aldehyde over 1,4‐dimethoxybenzene. Pollination by wind plus nocturnal pollinators (mainly moths) or by wind alone contributed less to seed set than pollination by wind plus diurnal pollinators (mainly bees). This suggests that the emission of scent during the night and attracting moths have no significant effect on reproductive success. It is possible that the emission of lilac aldehydes and other compounds at night is s result of phylogenetic constraints. Future studies should investigate whether moths may produce a marginal fitness gain in some years and/or some populations. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 624–640.     

杂交兰花色花香生物合成途径的转录组分析

该研究以杂交兰(Cymbidium hybrid)不同花色花香品种‘玉凤’(K18,黄色)和‘福韵丹霞’(K24,紫红色)为材料,采用RNA-Seq技术获得杂交兰不同花期的花朵转录组数据,分析杂交兰不同时期花色/花香相关基因的表达变化,探讨杂交兰花色花香形成的分子机理,为杂交的定向改良和新品种选育提供依据.结果表明:(...