Adaptation to pollination by fungus gnats underlies the evolution of pollination syndrome in the genus Euonymus.

BACKGROUND AND AIMS: Dipteran insects are known pollinators of many angiosperms, but knowledge on how flies affect floral evolution is relatively scarce. Some plants pollinated by fungus gnats share a unique set of floral characters (dark red display, flat shape and short stamens), which differs from any known pollination syndromes. We tested whether this set of floral characters is a pollination syndrome associated with pollination by fungus gnats, using the genus Euonymus as a model. METHODS: The pollinator and floral colour, morphology and scent profile were investigated for ten Euonymus species and Tripterygium regelii as an outgroup. The flower colour was evaluated using bee and fly colour vision models. The evolutionary association between fungus gnat pollination and each plant character was tested using a phylogenetically independent contrast. The ancestral state reconstruction was performed on flower colour, which is associated with fungus gnat pollination, to infer the evolution of pollination in the genus Euonymus. KEY RESULTS: The red-flowered Euonymus species were pollinated predominantly by fungus gnats, whereas the white-flowered species were pollinated by bees, beetles and brachyceran flies. The colour vision analysis suggested that red and white flowers are perceived as different colours by both bees and flies. The floral scents of the fungus gnat-pollinated species were characterized by acetoin, which made up >90 % of the total scent in three species. Phylogenetically independent contrast showed that the evolution of fungus gnat pollination is associated with acquisition of red flowers, short stamens and acetoin emission. CONCLUSIONS: Our results suggest that the observed combination of floral characters is a pollination syndrome associated with the parallel evolution of pollination by fungus gnats. Although the role of the red floral display and acetoin in pollinator attraction remains to be elucidated, our finding underscores the importance of fungus gnats as potential contributors to floral diversification.

Diversity and larval leaf-mining habits of Japanese jewel beetles of the tribe Tracheini (Coleoptera, Buprestidae).

From the Japanese Archipelago, 12 Habroloma and 20 Trachys species (Buprestidae: Tracheini) have been recorded. Two new Habroloma species were found, which are associated with Elaeocarpaceae and Loranthaceae, also new host plant families/orders for Tracheini. The two new species are described as Habrolomaelaeocarpusisp. nov. and Habrolomataxillusisp. nov., and the latter is the first Tracheini species shown to be associated with epiphytes. Leaf mines of 31 Tracheini species are also reported in this work, including new records of leaf mines for 16 Tracheini species. The larvae of all these recorded species are full-depth linear-blotch mesophyll miners of mature leaves and pupate within their mines. The mining habits of Habroloma species associated with Symplocos (Symplocaceae) are unique: the young larvae bore into midribs and petioles and cause leaf fall, and the larvae then mine the fallen leaves.

Erratum: Corrigendum: Kato M, Kawakita A (2023) Diversity and larval leaf-mining habits of Japanese jewel beetles of the tribe Tracheini (Coleoptera, Buprestidae). ZooKeys 1156: 133-158. doi: 10.3897/zookeys.1156.97768.

[This corrects the article DOI: 10.3897/zookeys.1156.97768.].

Phylogeny of gracillariid leaf-mining moths: evolution of larval behaviour inferred from phylogenomic and Sanger data.

Gracillariidae is the most taxonomically diverse cosmopolitan leaf-mining moth family, consisting of nearly 2000 named species in 105 described genera, classified into eight extant subfamilies. The majority of gracillariid species are internal plant feeders as larvae, creating mines and galls in plant tissue. Despite their diversity and ecological adaptations, their phylogenetic relationships, especially among subfamilies, remain uncertain. Genomic data (83 taxa, 589 loci) were integrated with Sanger data (130 taxa, 22 loci), to reconstruct a phylogeny of Gracillariidae. Based on analyses of both datasets combined and analyzed separately, monophyly of Gracillariidae and all its subfamilies, monophyly of the clade "LAMPO" (subfamilies: Lithocolletinae, Acrocercopinae, Marmarinae, Phyllocnistinae, and Oecophyllembiinae) and relationships of its subclade "AMO" (subfamilies: Acrocercopinae, Marmarinae, and Oecophyllembiinae) were strongly supported. A sister-group relationship of Ornixolinae to the remainder of the family, and a monophyletic leaf roller lineage (Callicercops Vári + Parornichinae) + Gracillariinae, as sister to the "LAMPO" clade were supported by the most likely tree. Dating analyses indicate a mid-Cretaceous (105.3 Ma) origin of the family, followed by a rapid diversification into the nine subfamilies predating the Cretaceous-Palaeogene extinction. We hypothesize that advanced larval behaviours, such as making keeled or tentiform blotch mines, rolling leaves and galling, allowed gracillariids to better avoid larval parasitoids allowing them to further diversify. Finally, we stabilize the classification by formally re-establishing the subfamily ranks of Marmarinae stat.rev., Oecophyllembiinae stat.rev. and Parornichinae stat.rev., and erect a new subfamily, Callicercopinae Li, Ohshima and Kawahara to accommodate the enigmatic genus Callicercops.

Three new species of Ametrodiplosis (Diptera: Cecidomyiidae) from Japan, with a key to the Japanese species and a molecular phylogenetic analysis.

Ametrodiplosis Rübsaamen (Diptera: Cecidomyiidae: Clinodiplosini) is a mostly Holarctic gall midge genus whose species are associated with a wide range of seed plant families, either as gall-inducers or inquilines. In this study, we describe three species of Ametrodiplosis from Japan: A. adetos n. sp. feeding in the flowers of Tylophora aristolochioides Miq. (Apocynaceae); A. aeroradicis n. sp. inducing aerial root galls on Trachelospermum asiaticum (Sieb. et Zucc.) Nakai and T. gracilipes var. liukiuense (Apocynaceae); and A. stellariae n. sp. forming leaf bud galls on Stellaria uliginosa Murray var. undulata (Thunb.) Ohwi (Caryophyllaceae). A molecular phylogenetic analysis using mitochondrial COI and ribosomal 16S genes and nuclear ribosomal 28S gene were conducted for the three new Ametrodiplisis species and other clinodiplosine taxa sequences available in GenBank. The analysis supported the monophyly of Ametrodiplosis despite the variable life history of the three species. In addition, it indicated very low intraspecific genetic divergence among the individuals from different localities and/or host plants. A taxonomic key to the three new Japanese species of Ametrodiplosis is provided.

Slippery flowers as a mechanism of defence against nectar-thieving ants.

BACKGROUND AND AIMS: The great diversity of floral characteristics among animal-pollinated plants is commonly understood to be the result of coevolutionary interactions between plants and pollinators. Floral antagonists, such as nectar thieves, also have the potential to exert an influence upon the selection of floral characteristics, but adaptation against floral antagonists has attracted comparatively little attention. We found that the corollas of hornet-pollinated Codonopsis lanceolata (Campanulaceae) and the tepals of bee-pollinated Fritillaria koidzumiana (Liliaceae) are slippery to nectar-thieving ants living in the plant's habitat; because the flowers of both species have exposed nectaries, slippery perianths may function as a defence against nectar-thieving ants. METHODS: We conducted a behavioural experiment and observed perianth surface microstructure by scanning electron microscopy to investigate the mechanism of slipperiness. Field experiments were conducted to test whether slippery perianths prevent floral entry by ants, and whether ant presence inside flowers affects pollination. KEY RESULTS: Scanning electron microscopy observations indicated that the slippery surfaces were coated with epicuticular wax crystals. The perianths lost their slipperiness when wiped with hexane. Artificial bridging of the slippery surfaces using non-slippery materials allowed ants to enter flowers more frequently. Experimental introduction of live ants to the Codonopsis flowers evicted hornet pollinators and shortened the duration of pollinator visits. However, no statistical differences were found in the fruit or seed sets of flowers with and without ants. CONCLUSIONS: Slippery perianths, most probably based on epicuticular wax crystals, prevent floral entry by ants that negatively affect pollinator behaviour. Experimental evidence of floral defence based on slippery surfaces is rare, but such a mode of defence may be widespread amongst flowering plants.

Active pollination drives selection for reduced pollen-ovule ratios.

PREMISE: Variation in pollen-ovule ratios is thought to reflect the degree of pollen transfer efficiency-the more efficient the process, the fewer pollen grains needed. Few studies have directly examined the relationship between pollen-ovule ratio and pollen transfer efficiency. For active pollination in the pollination brood mutualisms of yuccas and yucca moths, figs and fig wasps, senita and senita moths, and leafflowers and leafflower moths, pollinators purposefully collect pollen and place it directly on the stigmatic surface of conspecific flowers. The tight coupling of insect reproductive interests with pollination of the flowers in which larvae develop ensures that pollination is highly efficient. METHODS: We used the multiple evolutionary transitions between passive pollination and more efficient active pollination to test if increased pollen transfer efficiency leads to reduced pollen-ovule ratios. We collected pollen and ovule data from a suite of plant species from each of the pollination brood mutualisms and used phylogenetically controlled tests and sister-group comparisons to examine whether the shift to active pollination resulted in reduced pollen-ovule ratios. RESULTS: Across all transitions between passive and active pollination in plants, actively pollinated plants had significantly lower pollen-ovule ratios than closely related passively pollinated taxa. Phylogenetically corrected comparisons demonstrated that actively pollinated plant species had an average 76% reduction in the pollen-ovule ratio. CONCLUSIONS: The results for active pollination systems support the general utility of pollen-ovule ratios as indicators of pollination efficiency and the central importance of pollen transfer efficiency in the evolution of pollen-ovule ratio.

Leaf shape deters plant processing by an herbivorous weevil.

The shapes of plant leaves are remarkably diverse, but their ecological functions are largely unknown. Reports on the effects of leaf shape on biotic interactions such as herbivory are especially scarce, partly because herbivorous insects rarely rely on leaf shape for host selection. Here, we show that leaf shape acts as a physical deterrent against a leaf-processing herbivore. Plants in the genus Isodon (Lamiaceae) host a specialized leaf-rolling weevil (Apoderus praecellens) whose ovipositing females process an entire leaf into a leaf roll to serve as larval food and shelter. Among the species of Isodon, I. umbrosus var. hakusanensis is exceptional in that it has deeply lobed leaves. Because leaf processing follows a consistent sequence of complex behaviours, the unusual shape of I. umbrosus leaves may disrupt this process. Under both natural and laboratory conditions, female weevils preferred I. trichocarpus, a close relative with non-lobed leaves, over I. umbrosus. Nutritional properties of the leaves do not explain this preference because weevil larvae developed equally well on both hosts. Modifying the non-lobed I. trichocarpus leaves to mimic the shape of I. umbrosus leaves also discouraged leaf processing. Leaf processing often terminated because weevils failed to complete the inspection routine on I. umbrosus leaves. Leaf shape may be an important but overlooked factor that affects the interactions between plants and leaf-processing herbivores.

Leafflower-leafflower moth mutualism in the Neotropics: Successful transoceanic dispersal from the Old World to the New World by actively-pollinating leafflower moths.

In the Old World tropics, several hundred species of leafflowers (Phyllanthus sensu lato; Phyllanthaceae) are engaged in obligate mutualisms with species-specific leafflower moths (Epicephala; Gracillariidae) whose adults actively pollinate flowers and larvae consume the resulting seeds. Considerable diversity of Phyllanthus also exists in the New World, but whether any New World Phyllanthus is pollinated by Epicephala is unknown. We studied the pollination biology of four woody Phyllanthus species occurring in Peru over a period of four years, and found that each species is associated with a species-specific, seed-eating Epicephala moth, here described as new species. Another Epicephala species found associated with herbaceous Phyllanthus is also described. This is the first description of Epicephala from the New World. Field-collected female moths of the four Epicephala species associated with woody Phyllanthus all carried pollen on the proboscises, and active pollination behavior was observed in at least two species. Thus, Epicephala moths also pollinate New World Phyllanthus. However, not all of these Epicephala species may be mutualistic with their hosts, because we occasionally observed females laying eggs in developing fruits without pollinating. Also, the flowers of some Phyllanthus species were visited by pollen-bearing thrips or gall midges, which potentially acted as co-pollinators or primary pollinators. Phylogenetic analysis showed that the New World Epicephala associated with woody Phyllanthus are nested within lineages of Old World active pollinators. Thus, actively-pollinating Epicephala moths, which originated in the Old World, successfully colonized the New World probably across the Pacific and established mutualisms with resident Phyllanthus species, although whether any of the relationships are obligate requires further study. There is likely a major radiation of Epicephala still to be found in the New World.

Hawaiian Philodoria (Lepidoptera, Gracillariidae, Ornixolinae) leaf mining moths on Myrsine (Primulaceae): two new species and biological data.

This paper provides new taxonomic and biological data on a complex of gracillariid moths in the endemic genus Philodoria Walsingham, 1907 that are associated with Myrsine (Primulaceae) in the Hawaiian Islands, United States. Two new species, Philodoria kauaulaensis Kobayashi, Johns & Kawahara, sp. n. (host: Myrsine lanaiensis, M. lessertiana, and M. sandwicensis) and P. kolea Kobayashi, Johns & Kawahara, sp. n. (host: M. lessertiana) are described. Biological data are provided for two previously described species that also feed on Myrsine: P. auromagnifica Walsingham, 1907 and P. succedanea Walsingham, 1907. For the first time we detail and illustrate genital structures, immature stages, biology, and host plants of P. auromagnifica and P. succedanea. Philodoria kolea, P. auromagnifica, and P. succedanea occur in sympatry on the island of Hawaii (Big Island), but each species differs in behavioral characters: P. kolea utilizes leaves of seedlings and forms a serpentine mine, whereas the latter two utilize leaves of larger plants, and form linear or serpentine to blotch mines. More broadly, leaf mine forms and diagnostic characteristics of the Myrsine-feeding species complex of Philodoria (as currently known) are reviewed and illustrated.

Pollination by fungus gnats and associated floral characteristics in five families of the Japanese flora.

Background and aims: Pollination by fungus gnats (Mycetophilidae and Sciaridae) is uncommon, but is nevertheless known to occur in 20 genera among eight angiosperm families. Because many fungus gnat-pollinated plants possess a dark red floral display, we hypothesized that fungus gnat pollination is more widespread among plants with similar floral display than currently known. We thus studied the pollination biology of flowers with dark red pigmentation in five families, focusing particularly on plants having small, flat, actinomorphic flowers with exposed nectaries and short stamens, because these floral characteristics mirror those of a known fungus gnat-pollinated genus (Mitella). Methods: We observed daytime and night-time floral visitors for a total of 194.5 h in Aucuba japonica (Garryaceae), Euonymus spp. (Celastraceae), Disanthus cercidifolius (Hamamelidaceae), Micranthes fusca (Saxifragaceae) and Streptopus streptopoides (Liliaceae). Visitors were categorized into functional groups, and a pollination importance index (PII) was calculated for each functional group based on visitation frequency, pollen load and behaviour on flowers. Key results: Fungus gnats were dominant among the 1762 insects observed (36-92 % depending on the plant species) and were the most important pollinators among all plants studied (PII: 0.529-1). Fungus gnat visits occurred during the daytime and, more frequently, at dusk. Most often, pollen grains became clumped on the ventral side of the head and/or thorax as the short-proboscid fungus gnats foraged on nectar and came into contact with anthers located close to the flower base. Conclusions: Pollination by fungus gnats is probably more common than previously thought, especially in habitats similar to those of the plants studied (moist forest understorey, streamside or subalpine meadow) where fungus gnats are abundant year-round. Our results further suggest that there may be a previously unnoticed association between fungus gnat pollination and dark red coloration, and a shared overall floral architecture among the plants studied.

Limiting the cost of mutualism: the defensive role of elongated gynophore in the leafflower-moth mutualism.

Mutualisms are interactions from which both partners benefit but may collapse if mutualists' costs and benefits are not aligned. Host sanctions are one mechanism whereby hosts selectively allocate resources to the more cooperative partners and thereby reduce the fitness of overexploiters; however, many mutualisms lack apparent means of host sanctions. In mutualisms between plants and pollinating seed parasites, such as those between leafflowers and leafflower moths, pollinators consume subsets of the seeds as larval food in return for their pollination service. Plants may select against overexploiters by selectively aborting flowers with a heavy egg load, but in many leafflower species, seeds are fully eaten in some fruits, suggesting that such a mechanism is not present in all species. Instead, the fruits of Breynia vitis-idaea have stalk-like structures (gynophore) through which early-instar moth larvae must bore to reach seeds. Examination of moth mortality in fruits with different gynophore lengths suggested that fruits with longer gynophore had higher moth mortality and, therefore, less seed damage. Most moth mortality occurred at the egg stage or as early larval instar before moths reached the seeds, consistent with the view that gynophore functions to prevent moth access to seeds. Gynophore length was unaffected by plant size, extent of moth oviposition, or geography; thus, it is most likely genetically controlled. Because gynophores do not elongate in related species whose pollinators oviposit directly into the ovary, the gynophore in B. vitis-idaea may have evolved as a defense to limit the cost of the mutualism.

Pollinia transfer on moth legs in Hoya carnosa (Apocynaceae).

PREMISE OF THE STUDY: The genus Hoya (Apocynaceae; Asclepiadoideae) is characterized by a set of complex floral characters unique among the asclepiads, but their role in pollination is poorly understood. Here, we report a new mechanism of asclepiad pollination in the wax plant Hoya carnosa: the pollinaria are transferred on the legs of medium or large settling moths. METHODS: Floral visitors and their behavior were observed on Amami-Oshima Island during 2013-2015, and the efficacy of different pollinators was determined by counting the pollinarium loads on different flower visitors. The floral characters were studied to establish the process of pollination in relation to pollinator behavior on the flower. KEY RESULTS: Hoya carnosa was visited by various settling moths at night, but pollinia attachment was confirmed predominantly on the legs of the large moth Erebus ephesperis (Noctuidae) and less frequently on the legs of the medium-sized moths Bastilla arcuata (Crambidae) and Cleora injectaria (Geometridae). The moths walked around and searched for nectar on the inflorescence, and the corpusculum became clipped to the arolia (adhesive pads on moth tarsi) as they stepped on the pollinaria between the staminal corona. The downward spherical inflorescence of aggregated flowers with flat, velvety petals and a slippery corona provides restricted footholds for the visitors, which efficiently leads pollinator legs to the pollinaria. CONCLUSIONS: Although the pollination system of Hoya is largely unknown, pollination by insect legs may be a major pollination system in this genus because these basic floral characters are shared among many species.

Patterns of temporal and enemy niche use by a community of leaf cone moths (Caloptilia) coexisting on maples (Acer) as revealed by metabarcoding.

The diversity of herbivorous insects is often considered a function of host plant diversity. However, recent research has uncovered many examples of closely related herbivores using the same host plant(s), suggesting that partitioning of host plants is not the only mechanism generating diversity. Herbivores sharing hosts may utilize different parts of the same plant, but such resource partitioning is often not apparent; hence, the factors that allow closely related herbivores to coexist are still largely undetermined. We examined whether partitioning of phenology or natural enemies may explain the coexistence of leaf cone moths (Caloptilia; Gracillariidae) associated with maples (Acer; Sapindaceae). Larval activity of 10 sympatric Caloptilia species found on nine maple species was monitored every 2-3 weeks for a total of 13 sampling events, and an exhaustive search for internal parasitoid wasps was conducted using high-throughput sequencing. Blocking primers were used to facilitate the detection of wasp larvae inside moth tissue. We found considerable phenological overlap among Caloptilia species, with two clear peaks in July and September-October. Coexisting Caloptilia species also had largely overlapping parasitoid communities; a total of 13 chalcid and ichneumon wasp species attacked Caloptilia in a nonspecific fashion at an overall parasitism rate of 46.4%. Although coexistence may be facilitated by factors not accounted for in this study, it appears that niche partitioning is not necessary for closely related herbivores to stably coexist on shared hosts. Co-occurrence without resource partitioning may provide an additional axis along which herbivorous insects attain increased species richness.

Phylogenetic test of speciation by host shift in leaf cone moths (Caloptilia) feeding on maples (Acer).

The traditional explanation for the exceptional diversity of herbivorous insects emphasizes host shift as the major driver of speciation. However, phylogenetic studies have often demonstrated widespread host plant conservatism by insect herbivores, calling into question the prevalence of speciation by host shift to distantly related plants. A limitation of previous phylogenetic studies is that host plants were defined at the family or genus level; thus, it was unclear whether host shifts predominate at a finer taxonomic scale. The lack of a statistical approach to test the hypothesis of host-shift-driven speciation also hindered studies at the species level. Here, we analyze the radiation of leaf cone moths (Caloptilia) associated with maples (Acer) using a newly developed, phylogeny-based method that tests the role of host shift in speciation. This method has the advantage of not requiring complete taxon sampling from an entire radiation. Based on 254 host plant records for 14 Caloptilia species collected at 73 sites in Japan, we show that major dietary changes are more concentrated toward the root of the phylogeny, with host shift playing a minor role in recent speciation. We suggest that there may be other roles for host shift in promoting herbivorous insect diversification rather than facilitating speciation per se.

Revision of the Japanese species of Epicephala Meyrick with descriptions of seven new species (Lepidoptera, Gracillariidae).

Epicephala moths are involved in obligate mutualisms with their Phyllanthaceae hosts, in which the female moths assure pollination and, in return, their progeny develop by consuming the seeds. Ecological, molecular and geographical data suggest that the genus includes several hundred species, but the majority remains to be formally described. Here we revise the Japanese species of Epicephala Meyrick, 1880. In addition to two previously named species, seven species are newly described: Epicephala anthophilia sp. n., Epicephala lanceolatella sp. n., Epicephala perplexa sp. n., Epicephala obovatella sp. n., Epicephala corruptrix sp. n., Epicephala parasitica sp. n. and Epicephala nudilingua sp. n. The first four are species involved in obligate pollination mutualism, while the fifth is a pollinating seed parasite and the last two are derived non-pollinating seed parasites of herbaceous Phyllanthus. Each of the nine Japanese Epicephela species is specialized to a single plant species in the genera Glochidion, Breynia or Phyllanthus, except for Epicephala obovatella and Epicephala corruptrix that each utilizes two closely related Glochidion species. Considerable variations are found in pollination and oviposition behaviors among species, which are reflected in their proboscis and ovipositor morphologies, respectively. Molecular phylogeny indicated that there have been repeated transitions in oviposition mode during the diversification of Epicephala, which were accompanied by changes in ovipositor morphology, as suggested by a correlation analysis. Keys to species are provided.

Pollination system and the effect of inflorescence size on fruit set in the deceptive orchid Cephalanthera falcata.

Larger inflorescences in reward-producing plants can benefit plants by increasing both pollinator attraction and the duration of visits by individual pollinators. However, ultimately, inflorescence size is determined by the balance between the benefits of large inflorescences and the increased cost of geitonogamy. At present, little is known about the relationship between inflorescence size and fecundity in deceptive plants. Given that pollinators are likely to leave inflorescences lacking rewards quickly, it seems unlikely that longer pollinator visits and the risk of geitonogamy would be strong selective pressures in these species, which indicates that pollinator attraction might be the most important factor influencing their inflorescence size. Here we examined the pollination ecology of the deceptive orchid Cephalanthera falcata in order to clarify the effects of inflorescence size on the fruit set of this non-rewarding species. Field observations of the floral visitors showed that C. falcata is pollinated by the andrenid bee Andrena aburana, whilst pollination experiments demonstrated that this orchid species is neither autogamous nor apogamous, but is strongly pollinator dependent. Three consecutive years of field observations revealed that fruit set was positively correlated with the number of flowers per inflorescence. These results provide strong evidence that the nectarless orchid C. falcata benefits from producing larger inflorescences that attract a greater number of innate pollinators. Large inflorescences may have a greater positive effect on fruit set in deceptive plants because a growing number of studies suggest that fruit set in reward-producing plants is usually unaffected by display size.

Isolation and characterization of 11 microsatellite markers for Glochidion acuminatum (Phyllanthaceae).

PREMISE OF THE STUDY: Microsatellite markers were developed for Glochidion acuminatum (Phyllanthaceae) to investigate pollen dispersal distances and thereby to assess the effectiveness of specialized Epicephala moths as pollinators. • METHODS AND RESULTS: Using next-generation sequencing, 11 polymorphic microsatellite primer pairs were developed for G. acuminatum. The primer pairs were tested on 49 individuals from two populations in Japan. The number of alleles per locus ranged from two to 13, and the expected heterozygosity ranged from 0.12 to 0.85. Probability of identity for all loci combined was lower than 1.27 × 10(7). • CONCLUSIONS: The microsatellite markers developed in this study will be useful for evaluating the benefit of specialized Epicephala moth pollination to Glochidion plants.

Active pollination favours sexual dimorphism in floral scent.

Zoophilous flowers often transmit olfactory signals to attract pollinators. In plants with unisexual flowers, such signals are usually similar between the sexes because attraction of the same animal to both male and female flowers is essential for conspecific pollen transfer. Here, we present a remarkable example of sexual dimorphism in floral signal observed in reproductively highly specialized clades of the tribe Phyllantheae (Phyllanthaceae). These plants are pollinated by species-specific, seed-parasitic Epicephala moths (Gracillariidae) that actively collect pollen from male flowers and pollinate the female flowers in which they oviposit; by doing so, they ensure seeds for their offspring. We found that Epicephala-pollinated Phyllanthaceae plants consistently exhibit major qualitative differences in scent between male and female flowers, often involving compounds derived from different biosynthetic pathways. In a choice test, mated female Epicephala moths preferred the scent of male flowers over that of female flowers, suggesting that male floral scent elicits pollen-collecting behaviour. Epicephala pollination evolved multiple times in Phyllantheae, at least thrice accompanied by transition from sexual monomorphism to dimorphism in floral scent. This is the first example in which sexually dimorphic floral scent has evolved to signal an alternative reward provided by each sex, provoking the pollinator's legitimate altruistic behaviour.