Pollinator-Mediated Selection on Flower Color,Flower Scent and Flower Morphology of Hemerocallis: Evidence from Genotyping Individual Pollen Grains On the Stigma

To trace the fate of individual pollen grains through pollination processes, we determined genotypes of single pollen grains deposited on Hemerocallis stigmas in an experimental mixed-species array. Hemerocallis fulva, pollinated by butterflies, has diurnal, reddish and unscented flowers, and H. citrina, pollinated by hawkmoths, has nocturnal, yellowish and sweet scent flowers. We observed pollinator visits to an experimental array of 24 H. fulva and 12 F2 hybrids between the two species (H. fulva and H. citrina) and collected stigmas after every trip bout of swallowtail butterflies or hawkmoths. We then measured selection by swallowtail butterflies or hawkmoths through male and female components of pollination success as determined by single pollen genotyping. As expected, swallowtail butterflies imposed selection on reddish color and weak scent: the number of outcross pollen grains acquired is a quadratic function of flower color with the maximum at reddish color, and the combined pollination success was maximal at weak scent (almost unrecognizable for human). This explains why H. fulva, with reddish flowers and no recognizable scent, is mainly pollinated by swallowtail butterflies. However, we found no evidence of hawkmoths-mediated selection on flower color or scent. Our findings do not support a hypothesis that yellow flower color and strong scent intensity, the distinctive floral characteristics of H. citrina, having evolved in adaptations to hawkmoths. We suggest that the key trait that triggers the evolution of nocturnal flowers is flowering time rather than flower color and scent.     

UV bullseye contrast of Hemerocallis flowers attracts hawkmoths but not swallowtail butterflies

The color and patterns of animal‐pollinated flowers are known to have effects on pollinator attraction. In this study, the relative importance of flower color and color contrast patterns on pollinator attraction was examined in two pollinator groups, swallowtail butterflies and hawkmoths using two Hemerocallis species; butterfly‐pollinated H. fulva and hawkmoth‐pollinated H. citrina, having reddish and yellowish flowers in human vision, respectively. Flowers of both species have UV bullseye patterns, composed of UV‐absorbing centers and UV‐reflecting peripheries, known to function as a typical nectar guide, but UV reflectance was significantly more intense in the peripheries of H. citrina flowers than in those of H. fulva flowers. Comparison based on the visual systems of butterflies and hawkmoths showed that the color contrast of the bullseye pattern in H. citrina was more intense than that in H. fulva. To evaluate the relative importance of flower color and the color contrast of bullseye pattern on pollinator attraction, we performed a series of observations using experimental arrays consisting of Hemerocallis species and their hybrids. As a result, swallowtail butterflies and crepuscular/nocturnal hawkmoths showed contrasting preferences for flower color and patterns: butterflies preferred H. fulva‐like colored flower whereas the preference of hawkmoths was affected by the color contrast of the bullseye pattern rather than flower color. Both crepuscular and nocturnal hawkmoths consistently preferred flowers with stronger contrast of the UV bullseye pattern, whereas the preference of hawkmoths for flower color was incoherent. Our finding suggests that hawkmoths can use UV‐absorbing/reflecting bullseye patterns for foraging under light‐limited environments and that the intensified bullseye contrast of H. citrina evolved as an adaptation to hawkmoths. Our results also showed the difference of visual systems between pollinators, which may have promoted floral divergence.     

Variation of flower opening and closing times in F1 and F2 hybrids of daylily (Hemerocallis fulva; Hemerocallidaceae) and nightlily (H. citrina)

In flowering plants, pollination success is strongly dependent on the timing of when flowers start to bloom and when they start to close. To elucidate the genetic mechanism influencing the timing of flower opening and closing, we obtained F1 and F2 hybrids of Hemerocallis fulva (a diurnally blooming species, pollinated by swallowtail butterflies) and H. citrina (a nocturnally blooming species, pollinated by nocturnal hawkmoths) and observed their flowering behavior from blooming to closing with the use of digital cameras. For flower opening times, F1 hybrids were highly variable, and F2 hybrids showed a bimodal distribution of flower opening times with peaks in both the morning and evening. The ratio of morning flowering and evening flowering among F2 hybrids did not deviate from 1:1. For the start to close time, both F1 and F2 hybrids were similar in showing the major peak in the evening. The ratio of evening closing and morning closing among F2 hybrids did not deviate from 3:1. These results suggest that the time of flower opening and the start of closing are regulated by different major genes.     

Day–night fluctuations in floral scent and their effects on reproductive success in <Emphasis Type="Italic">Lilium auratum</Emphasis>

We examined the contribution of diurnal and nocturnal pollination to male and female reproductive success in Lilium auratum. Plants were bagged for either 12 h during the day or at night to allow either only nocturnal or only diurnal visitors to forage throughout the flowering period. We found that there was no significant difference in the seed:ovule ratio among diurnally pollinated, nocturnally pollinated, or control flowers. However, in terms of male reproductive success, it was more advantageous for the plants to be pollinated both diurnally and nocturnally: the numbers of pollen grains remaining in diurnally pollinated or nocturnally pollinated flowers were significantly greater than those in control flowers. The total amount of floral volatiles of L. auratum was significantly higher at night than during the day. The constituents of floral scent of all time series examined were mostly monoterpenoids, many of which serve as attractants for nocturnal hawkmoths. Such nocturnally biased floral scent emission of L. auratum might achieve male reproductive success by attracting nocturnal visitors, which may suggest that the relative contribution of floral scent in this species is biased towards male reproductive success.     

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.     

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.     

Pollination biology of Lilium japonicum var. abeanum and var. japonicum: evidence of adaptation to the different availability of diurnal and nocturnal pollinators

Floral traits such as color and size are highly diversified in lilies, but their adaptive significance remains uncertain. In the present study, we compared pollination processes between Lilium japonicum var. abeanum and var. japonicum to clarify how the two varieties are adapted to different pollinators. Lilium japonicum var. japonicum is known to be pollinated by moths, and we hypothesized from its flower traits that var. abeanum is pollinated by diurnal insects. Using waterproof digital cameras set to a recording interval of 10–30 s, we recorded flower visitors for 7–9 flowers of var. japonicum and 4–6 flowers of var. abeanum over 24‐h periods. We also recorded the number of versatile (T‐shaped) and rigid (I‐shaped) anthers per flower. For var. abeanum, we observed flowers at 05.00 and 17.00 hours to determine flower opening time and measured the intensity of floral scent using a metal‐oxide semi‐conductor odor sensor. Both diurnal and nocturnal insects visited flowers of the two varieties, but visitation of diurnal insects was more frequent in var. abeanum. Anthers of var. abeanum are usually rigid, as in many bee‐pollinated flowers, whereas those of var. japonicum are mostly versatile. Although flowers of var. japonicum are known to open in the evening, 32% of the flowers of var. abeanum started to flower during the day. Lilium japonicum var. abeanum emitted scent at night, with a maximum intensity at 20.00 hours, just as in var. japonicum. These findings suggest that the floral traits of the two varieties are adapted to the different relative availabilities of nocturnal versus diurnal pollinators. The flower of var. abeanum is more adapted to diurnal pollinators, but retains adaptations to nocturnal pollinators, particularly with regard to its scent.     

Intrafloral patterns of color and scent in Capparis spinosa L. and the ghosts of its selection past

Premise

Capparis spinosa is a widespread charismatic plant, in which the nocturnal floral habit contrasts with the high visitation by diurnal bees and the pronounced scarcity of hawkmoths. To resolve this discrepancy and elucidate floral evolution of C. spinosa, we analyzed the intrafloral patterns of visual and olfactory cues in relation to the known sensory biases of the different visitor guilds (bees, butterflies, and hawkmoths).

Methods

We measured the intrafloral variation of scent, reflectance spectra, and colorimetric properties according to three guilds of known visitors of C. spinosa. Additionally, we sampled visitation rates using a motion-activated camera.

Results

Carpenter bees visited the flowers eight times more frequently than nocturnal hawkmoths, at dusk and in the following morning. Yet, the floral headspace of C. spinosa contained a typical sphingophilous scent with high emission rates of certain monoterpenes and amino-acid derived compounds. Visual cues included a special case of multisensory nectar guide and color patterns conspicuous to the visual systems of both hawkmoths and bees.

Conclusions

The intrafloral patterns of sensory stimuli suggest that hawkmoths have exerted strong historical selection on C. spinosa. Our study revealed two interesting paradoxes: (a) the flowers phenotypically biased towards the more inconsistent pollinator; and (b) floral display demands an abundance of resources that seems maladaptive in the habitats of C. spinosa. The transition to a binary pollination system accommodating large bees has not required phenotypic changes, owing to specific eco-physiological adaptations, unrelated to pollination, which make this plant an unusual case in pollination ecology.     

Context-dependent reproductive isolation mediated by floral scent and color

Reproductive isolation due to pollinator behavior is considered a key mode of speciation in flowering plants. Although floral scent is thought to mediate pollinator behavior, little is known about its effects on pollinator attraction and floral visitation in the wild. We used field experiments with wild hawkmoths and laboratory experiments with naïve hawkmoths to investigate attraction to and probing of flowers in response to indole, a volatile emitted by Ipomopsis tenuituba but not its close relative I. aggregata, both alone and in combination with floral color differences. We demonstrated that indole attracts wild hawkmoths to flowers, but has little effect on the rate at which those attracted moths probe flowers. In contrast, white flower color did not influence hawkmoth attraction in the field, but caused more attracted moths to probe flowers. Thus, the moths require both scent and high visual contrast, in that order, to feed at flowers at dusk. Their preference for indole-scented flowers is innate, but species-specific preference is mitigated by previous experience and plant spatial patterning. This context-dependent behavior helps explain why these Ipomopsis species show geographical variation in the extent of hybridization and may potentially explain formation of hybrid bridges in other systems of hawkmoth-pollinated plants.     

Beyond the pollination syndrome: nectar ecology and the role of diurnal and nocturnal pollinators in the reproductive success of Inga sessilis (Fabaceae)

Inga species present brush‐type flower morphology allowing them to be visited by distinct groups of pollinators. Nectar features in relation to the main pollinators have seldom been studied in this genus. To test the hypothesis of floral adaptation to both diurnal and nocturnal pollinators, we studied the pollination ecology of Inga sessilis, with emphasis on the nectar secretion patterns, effects of sequential removals on nectar production, sugar composition and the role of diurnal and nocturnal pollinators in its reproductive success. Inga sessilis is self‐incompatible and pollinated by hummingbirds, hawkmoths and bats. Fruit set under natural conditions is very low despite the fact that most stigmas receive polyads with sufficient pollen to fertilise all ovules in a flower. Nectar secretion starts in the bud stage and flowers continually secreting nectar for a period of 8 h. Flowers actively reabsorbed the nectar a few hours before senescence. Sugar production increased after nectar removal, especially when flowers were drained during the night. Nectar sugar composition changed over flower life span, from sucrose‐dominant (just after flower opening, when hummingbirds were the main visitors) to hexose‐rich (throughout the night, when bats and hawkmoths were the main visitors). Diurnal pollinators contributed less than nocturnal ones to fruit production, but the former were more constant and reliable visitors through time. Our results indicate I. sessilis has floral adaptations, beyond the morphology, that encompass both diurnal and nocturnal pollinator requirements, suggesting a complementary and mixed pollination system.     

Character displacement in style length between pollinator-sharing Clerodendrum trichotomum and C. izuinsulare (Verbenaceae)

Competition for pollination may occur between pollinator-sharing sympatric plants and this may cause character displacement of their floral traits. We examined this possibility by comparing flower morphology of the sympatric population of Clerodendrum trichotomum and its co-flowering congener, C. izuinsulare, with that of the allopatric populations. The two species were visited in common by such insects as diurnal hawkmoths, bees, swallowtails and nocturnal hawkmoths, and were pollinated nocturnally as well as diurnally. Interspecific pollen transfer can occur by sharing pollinators; however, they did not hybridize when artificially pollinated. Flower size, including stamen and style lengths, is larger in C. trichotomum with an overlap in range. The style of C. izuinsulare in the sympatric population was significantly shorter than that in the allopatric population, while there was no significant difference in style length between the allopatric and the sympatric C. trichotomum. This seems to facilitate avoidance of interspecific pollen transfer in the sympatric population.Ken Inoue was killed in an accident on July 28, 2003, during his research trip in Sakhalin.     

Generalized pollination system: Are floral traits adapted to different pollinators?

Flowers that are pollinated both during the day and at night could exhibit two different groups of pollinators and produce two different sets of attractants and rewards. We explored the patterns of emission of flower scents and production of nectar in the cactus Echinopsis chiloensis ssp. chiloensis, in relation to the patterns of activity of its diurnal and nocturnal pollinators. We measured frequency of flower visitors, analyzed floral scents, measured nectar production and sugar concentration, and performed pollination exclusion experiments. Bees were the main visitors at daytime and hawkmoths at nighttime. Diurnal scents were dominated by several compounds that can attract a wide range of pollinators, whereas nocturnal scents were less diverse and were dominated by (E)-nerolidol, a compound eliciting antennal responses in hawkmoths. Nectar volume and sugar concentration at night were similar to those recorded in hawkmoth-pollinated flowers. Daytime nectar volume was higher than those commonly found in bee-pollinated flowers, but similar to those found in flowers pollinated by several pollinators. Daytime sugar concentration was similar to those recorded in bee-pollinated flowers. Flowers of E. chiloensis ssp. chiloensis seem morphologically adapted to hawkmoth pollination, but diurnal and nocturnal pollinators contribute to similar extents to reproductive success. Additionally, diurnal and nocturnal pollinators showed a synergic effect on the product of fruit set and seed set. The results are discussed in terms of the linkage between floral traits and perception abilities and requirements of pollinators.     

Pollination of <Emphasis Type="Italic">Machaerium opacum</Emphasis> (Fabaceae) by nocturnal and diurnal bees

With plants whose flowers open at night and stay open during the day, nocturnal pollinators may exploit floral resources before diurnal competitors. Moths, bats, and beetles are the most familiar nocturnal pollinators, whereas nocturnal bees as pollinators remain poorly understood. The common Cerrado tree Machaerium opacum (Fabaceae) has white and strongly scented melittophilous flowers, which first open at the night and remain open during the day and, thus, have the potential to be visited by both nocturnal and diurnal bees. We asked: (1) what is the plant’s breeding system? (2) when during the night do the flowers open? (3) what are the visual and olfactory floral cues? and (4) which nocturnal/diurnal bees visit and pollinate the flowers? We show that M. opacum is self-incompatible. Its flowers open synchronously at 03:30 h, produce nectar exclusively at night, and have an explosive mechanism of pollen presentation. The flowers have pure white petals, release strong scents during anthesis, and are pollinated by nocturnal and diurnal bees. We recorded four nocturnal and 17 diurnal species as flower visitors, with females of nocturnal species of Ptiloglossa (Colletidae) being the most abundant. After an initial pollen-releasing visit, only a minor amount of pollen remains in a flower. Several floral traits favor visits by nocturnal bees: (1) night-time flower opening, (2) nectar production at night, (3) almost complete pollen release during the first flower visit, and (4) pure white petals and strong odor production prior to sunrise, facilitating visual and olfactory detection of flowers when light is dim.     

Fitness consequences of altering floral circadian oscillations for Nicotiana attenuata

Ecological interactions between flowers and pollinators are all about timing.Flower opening/closing and scent emissions are largely synchronized with pollinator activity,and a circadian clock regulates these rhythms.However,whether the circadian clock increases a plant's reproductive success by regulating these floral rhythms remains untested.Flowers of Nicotiana attenuata,a wild tobacco,diurnally and rhythmically open,emit scent and move vertically through a 140° arc to interact with nocturnal hawkmoths.We tethered flowers to evaluate the importance of flower positions for Manduca sexta-mediated pollinations;flower position dramatically influenced pollination.We examined the pollination success of phase-shifted flowers,silenced in circadian clock genes,NaZTL,NaLHY,and NaTOCi,by RNAi.Circadian rhythms in N.attenuata flowers are responsible for altered seed set from outcrossed pollen.     

Pollination in Lilium sargentiae (Liliaceae) and the first confirmation of long‐tongued hawkmoths as a pollinator niche in Asia

The geographical distribution of pollinators is an important factor determining the biogeographical pattern of floral evolution and diversification. In Africa, a guild of plant species has converged in a floral syndrome for pollination by long‐tongued hawkmoths (predominantly Agrius convolvuli). It is hypothesized that such floral convergence could track the geographical distribution of long‐tongued hawkmoths, so it may not be confined to Africa. We investigated the pollination biology of Lilium sargentiae E. H. Wilson, which is endemic to China and exhibits traits suggestive of long‐tongued hawkmoth pollination. Lilium sargentiae was visited by A. convolvuli as well as small beetles (Nitidulidae) but pollination was mostly or totally effected by the moth. It was consistent with other hawkmoth‐pollinated plants in terms of floral tube length, nectar traits, tepal reflectance, and scent composition. We present the first experimental evidence for the hypothesis proposed above and for hawkmoth pollination in the widespread and ornamentally and economically important genus Lilium L. in a natural habitat. Our findings imply that long‐tongued hawkmoths (especially the extremely widespread A. convolvuli) as a distinctive pollinator niche may have underlain the evolution of long‐tubed (>8 cm) flowers across different continents.     

Floral scent emission is the highest at the second night of anthesis in Lonicera japonica (Caprifoliaceae)

Floral color change in diverse plants has been thought to be a visual signal reflecting changes in floral rewards, promoting pollinator foraging efficiency as well as plant reproductive success. It remains unclear whether olfactory signals co-vary with floral color change. We investigated the production rhythms of floral scent and nectar associated with floral color change in Lonicera japonica. The flowers generally last 2–3 days. They are white on opening at night (N1) and become light yellow the following day (D1), yellow on the second night (N2), and golden on the second day of flowering (D2). Our measurements in the four stages indicated that nectar production decreased significantly from N1 and D1 to N2 and D2, tracking the floral color change. A total of 34 compounds were detected in floral scent and total scent emission was significantly higher in N2 than in the other three stages. The scent emission of three major compounds, Linalool, cis-3-Hexenyl tiglate, and Germacrene D was also significantly higher in N2, but the relative content of Linalool decreased gradually, cis-3-Hexenyl tiglate increased gradually, and the relative content of Germacrene D did not differ among the four measured stages. Greater scent emission by night than by day suggested a strong olfactory signal to attract nocturnal hawkmoths, the effective pollinators. However, floral scent rhythms in the four stages did not match the color change and nectar secretion, suggesting that floral color (visual) and scent (olfactory) in this species may play different roles in attracting or filtering various visitors.     

Exploring the boundary between pollination syndromes: bats and hummingbirds as pollinators of<Emphasis Type="Italic"> Burmeistera cyclostigmata</Emphasis> and<Emphasis Type="Italic"> B. tenuiflora</Emphasis> (Campanulaceae)

In this study I documented the degree of specialization in the pollination systems of Burmeistera cyclostigmata and B. tenuiflora (Campanulaceae) to explore the potential role of floral isolation in the diversification of the genus. I asked which floral characteristics are important in specializing on either bat or hummingbird pollination, and whether overlap between these floral syndromes can exist. I examined nocturnal and diurnal pollen deposition, pollinator visitation rates, and single visit effectiveness and related them to intra- and interspecific variation in Burmeistera floral characteristics at Monteverde, Costa Rica. Bats and hummingbirds visited both Burmeistera species, and bats pollinated both species. Owing to differences in floral morphology, however, hummingbirds effectively pollinated only B. tenuiflora. The generalized pollination system of B. tenuiflora demonstrates that there can be overlap in the boundary between ornithophily and chiropterophily, and that nectar production and timing of anthesis do not serve as barriers between these syndromes. The high intraspecific variation in floral color from green to red or purple did not correlate with either nocturnal or diurnal pollen deposition. Degree of flower accessibility did affect pollination; nocturnal pollen deposition significantly decreased as flowers become more obstructed. In Burmeistera, floral morphology and accessibility appear to be the most important floral characteristics for specialization at the boundary between ornithophily and chiropterophily.     

Pollination ecology of Agave macroacantha (Agavaceae) in a Mexican tropical desert. I. Floral biology and pollination mechanisms

In a study of sexual reproduction in long-lived semelparous plants, we observed Agave macroacantha in the tropical desert of Tehuacán-Cuicatlán, Mexico, describing duration of flowering, flower phenology, and nectar production patterns. We also performed two manipulative experiments evaluating (a) the seed production efficiency of different crossing systems (selfing, cross-pollination, apomixis, and control), and (b) the effect of different pollinators (diurnal exposure to pollinators, nocturnal exposure, exclusion, and control) on the seeds produced. Flowering occurred from early May to late July and had a mean duration of 29 days in the individual rosettes. The flowers were protandrous; anthesis occurred in the afternoon of the third day after floral opening, and the pistils matured in the afternoon of the fifth day. The stigmas remained receptive from dusk to the following morning. Pollination was mostly allogamous. Nectar was produced principally during the night, from the first stages of floral aperture until the stigmas wilted and flowering ceased. The flowers were visited during the day by hymenoptera, butterflies, and hummingbirds and during the night by bats and moths. Only the nocturnal visitors, however, were successful pollinators. Agave macroacantha is extremely dependent on nocturnal pollinators for its reproductive success.     

Nectar Production of Calliandra longipedicellata (Fabaceae: Mimosoideae), an Endemic Mexican Shrub with Multiple Potential Pollinators

The nectar resource environment across which nectarivores forage may be patchy and variable. To understand the sources and consequences of such a variation, nectar production was investigated in Calliandra longipedicellata . Nectar was measured once a month throughout a 3-mo winter season in two successive years at three sites. We also conducted diurnal and nocturnal field observations to describe visitation rates of floral visitors, and a pollinator exclusion experiment to evaluate diurnal and nocturnal pollination at the three sites. In all populations, nectar secretion was primarily nocturnal, although flowers produced some nectar during the day. Sugar production per flower varied significantly at both the seasonal and population levels, although nectar production rates and a well-defined afternoon to morning production pattern were consistent across months, populations, and years. Average nectar production rates were high compared to other Calliandra species, and to most hummingbird- or hawkmoth-pollinated plants in the region. Flowers were regularly visited by hawkmoths, bats, hummingbirds and various diurnal insects, and all populations had similar rates of visitation. Nocturnal insects had the highest overall visitation rates (three times as high as those by diurnal insects). Fruit production varied among pollination treatments and populations, and significant differences were found in fruit production when flowers exposed to both diurnal and nocturnal visitation were compared to flowers exposed only to diurnal visitation. Our results and the bright-red staminal filaments of C. longipedicellata indicate lack of specialization for particular pollinators.     

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.