The importance of oligosulfides in the attraction of fly pollinators to the brood-site deceptive species Jaborosa rotacea (Solanaceae).

Premise of research. Brood-site deceptive flowers use dishonest signals?especially floral odors that mimic oviposition substrates?to attract and deceive saprophilous insects to pollinate them. In this work, we recorded the pollinators of the sapromyiophilous species Jaborosa rotacea (Solanaceae) endemic to southern South America. Then, we characterized the floral volatiles of this species, and finally, we carried out field experiments to decouple the effects of scent and color as attractants for saprophilous flies. Methodology. We made direct observations of pollinators in a natural population of J. rotacea.We characterized floral volatiles by means of gas chromatography?mass spectrometry. Subsequently, we used a mixture of 2 oligosulfides (dimethyl disulfide and dimethyl trisulfide), which our analyses revealed were the main constituents of the floral scent ofJ. rotacea, as baits to determine the attractiveness of this olfactory signal to flies in a geographical region whereJ. rotaceais not present. Finally, we used the same foul-scented baits in arrays of artificial flowers resembling those of J. rotacea to assess the dual importance of olfactory and visual cues in fly attraction. Pivotal results. Pollination of J. rotaceaoccurs when saprophilous flies belonging to the families Calliphoridae, Muscidae, and Sarcophagidae?with similar body dimensions to the anther-stigma distance in these flowers?acquire and deposit pollen in the flowers in a nototribic mode. Our chemical analyses revealed that J. rotacea floral scent is chemically simple and features 2 oligosulfide compounds (dimethyl disulfide and dimethyl trisulfide) commonly found in carrion-mimicking flowers. We found that saprophilous flies belonging to the same families that we recorded as pollinators of J. rotaceain its native South American habitat were attracted to foul-scented baits in temperate North America. The flies? visitation frequencies (recorded as approaches and landings on the artificial flowers) depended significantly on the presence of the foul-scented baits. Conclusions. These results support the hypothesis that oligosulfides are universally effective signals by which deceptive flowers may effect pollen dispersal by attracting flies that use carrion or carnivore feces as brood sites. Keywords:brood-site deceptive flowers, Diptera, Jaborosa rotacea, oligosulfides, scent mimicry, Solanaceae.Fil: More, Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); Argentina;Fil: Cocucci, Andrea Aristides. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal (p); Argentina;Fil: Raguso, Robert A.. Cornell University; Estados Unidos de América

Olfactory versus visual cues in a floral mimicry system

 We used arrays of artificial flowers with and without fragrance to determine the importance of olfactory and visual cues in attracting insects to a floral mimic. The mimic is a fungus, Puccinia monoica Arth., which causes its crucifer hosts (here, Arabis drummondii Gray) to form pseudoflowers that mimic co-occurring flowers such as the buttercup, Ranunculus inamoenus Greene. Although pseudoflowers are visually similar to buttercups, their sweet fragrance is distinct. To determine whether visitors to pseudoflowers were responding to fragrance we performed an experiment in which we removed the visual cues, but allowed fragrance to still be perceived. In this experiment we found that pseudoflower fragrance can attract visitors by itself. In other experiments we found that the relative importance of olfactory and visual cues depended on the species of visitor. Halictid bees ( Dialictus sp.) had a somewhat greater visual than olfactory response, whereas flies (muscids and anthomyiids) were more dependent on olfactory cues. We also used bioassays to determine which of the many compounds present in the natural fragrance were responsible for attraction. We found that halictid bees were equally attracted to pseudoflowers and to a blend containing phenylacetaldehyde, 2-phenylethanol, benzaldehyde and methylbenzoate in the same relative concentrations as in pseudoflowers. Flies, on the other hand, only responded to pseudoflower scent, indicating that we have not yet identified the compound(s) present in pseudoflowers that are attracting them. The ability of insects to differentiate pseudoflowers from true flowers by their fragrance may be important in the evolution of the mimicry system. Different fragrances may facilitate proper transfer of both fungal spermatia and pollen, and thus make it possible for the visual mimicry to evolve.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/42277/1/442-109-3-414_71090414.pd

Electroantennogram responses of male Sphinx perelegans hawkmoths to floral and ‘green-leaf volatiles’

Electroantennograms (EAGs) from field-collected male Sphinx perelegans hawkmoths were recorded in response to 10 individual floral scent compounds identified from Clarkia breweri (Onagraceae), 21 additional volatiles characteristic of other night-blooming flowers, and eight ‘green leaf’ volatiles. Measurable EAG responses were elicited to all compounds tested, but the most effective antennal stimulants were benzyl acetate, linalool, methyl salicylate and trans-2-hexenal. Mean, pooled EAGs to oxygenated terpenoids, aromatic esters and fatty acid derivatives were larger in magnitude than those in response to aromatic aldehydes/alcohols, monoterpenes and nitrogen-bearing compounds. The rank order of male S. perelegans' EAGs did not differ significantly from that of previously recorded responses of male Hyles lineata to the same scent compounds, and EAG magnitudes were generally larger for S. perelegans than for H. lineata. Both hawkmoth species are shown to have broad olfactory receptivities and could potentially respond to a wide array of plant volatiles as floral attractants.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/42717/1/10667_2004_Article_158733.pd

Soil fungal effects on floral signals, rewards, and aboveground interactions in an alpine pollination web

This is the published version. The definitive version is available at www3.interscience.wiley.com.• Premise of the study: Plants interact with above- and belowground organisms; the combined effects of these interactions determine plant fitness and trait evolution. To better understand the ecological and evolutionary implications of multispecies interactions, we explored linkages between soil fungi, pollinators, and floral larcenists in Polemonium viscosum (Polemoniaceae). • Methods: Using a fungicide, we experimentally reduced fungal colonization of krummholz and tundra P. viscosum in 2008–2009. We monitored floral signals and rewards, interactions with pollinators and larcenists, and seed set for fungicide-treated and control plants. • Key results: Fungicide effects varied among traits, between interactions, and with environmental context. Treatment effects were negligible in 2008, but stronger in 2009, especially in the less-fertile krummholz habitat. There, fungicide increased nectar sugar content and damage by larcenist ants, but did not affect pollination. Surprisingly, fungicide also enhanced seed set, suggesting that direct resource costs of soil fungi exceed indirect benefits from reduced larceny. In the tundra, fungicide effects were negligible in both years. However, pooled across treatments, colonization by mycorrhizal fungi in 2009 correlated negatively with the intensity and diversity of floral volatile organic compounds, suggesting integrated above- and belowground signaling pathways. • Conclusions: Fungicide effects on floral rewards in P. viscosum link soil fungi to ecological costs of pollinator attraction. Trait-specific linkages to soil fungi should decouple expression of sensitive and buffered floral phenotypes in P. viscosum. Overall, this study demonstrates how multitrophic linkages may lead to shifting selection pressures on interaction traits, restricting the evolution of specialization.National Science Foundation (DBI-0603049 and DEB-0316110 to C.G. and DEB-0746106 to R.A.R.)

Flower diversification across “pollinator climates”: Sensory aspects of corolla color evolution in the florally diverse south american genus Jaborosa (Solanaceae)

Flower phenotype may diverge within plant lineages when moving across “pollinator climates” (geographic differences in pollinator abundance or preference). Here we explored the potential importance of pollinators as drivers of floral color diversification in the nightshade genus Jaborosa, taking into account color perception capabilities of the actual pollinators (nocturnal hawkmoths vs. saprophilous flies) under a geographic perspective. We analyzed the association between transitions across environments and perceptual color axes using comparative methods. Our results revealed two major evolutionary themes in Jaborosa: (1) a “warm subtropical sphingophilous clade” composed of three hawkmoth-pollinated species found in humid lowland habitats, with large white flowers that clustered together in the visual space of a model hawkmoth (Manduca sexta) and a “cool-temperate brood-deceptive clade” composed of largely fly-pollinated species with small dark flowers found at high altitudes (Andes) or latitudes (Patagonian Steppe), that clustered together in the visual space of a model blowfly (Lucilia sp.) and a syrphid fly (Eristalis tenax). Our findings suggest that the ability of plants to colonize newly formed environments during Andean orogeny and the ecological changes that followed were concomitant with transitions in flower color as perceived by different pollinator functional groups. Our findings suggest that habitat and pollination mode are inextricably linked in the history of this South American plant lineage.Fil: Moré, Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Ibañez, Ana Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Drewniak, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Cocucci, Andrea Aristides. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Raguso, Robert A.. Cornell University; Estados Unido

Breakdown of species boundaries in Mandevilla: Floral morphological intermediacy, novel fragrances and asymmetric pollen flow

Phenotypic intermediacy is an indicator of putative hybrid origin, and has provided the main clues to discovering hybrid plants in nature. Mandevilla pentlandiana and M. laxa (Apocynaceae) are sister species with clear differences in floral phenotype and associated pollinator guilds: diurnal Hymenoptera and nocturnal hawkmoths, respectively. The presence of individuals with intermediate phenotypes in a wild population raises questions about the roles of visual and olfactory signals (i.e. corolla morphology and floral fragrances) as barriers to interbreeding, and how the breakdown of floral isolation occurs.We examined phenotypic variation in a mixed Mandevilla population analysing the chemical composition of floral fragrances, characterizing floral shape through geometric morphometrics and assessing individual grouping through taxonomically relevant traits and an unsupervised learning algorithm. We quantified the visitation frequencies of floral visitors and tracked their foraging movements using pollen analogues.The presence of morphologically intermediate individuals and pollen analogue movement suggested extensive hybridization between M. laxa and M. pentlandiana, along with asymmetrical rates of backcrossing between these putative hybrids and M. laxa. Floral volatiles from putative hybrid individuals showed a transgressive phenotype, with additional compounds not emitted by either parental species.Our results suggest the presence of a hybrid swarm between sympatric M. pentlandiana and M. laxa and indicate that initial hybridization events between these parental species are rare, but once they occur, visits between putative hybrids and M. laxa are common and facilitate continued introgression.Fil: Rubini Pisano, María Aimé. Universidad Nacional Autónoma de México; México. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: More, Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Cisternas, Mauricio. Jardín Botánico Nacional; Chile. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Raguso, Robert A.. Cornell University; Estados UnidosFil: Benitez-Vieyra, Santiago Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentin

Geographic variation in floral scent of Echinopsis ancistrophora (Cactaceae); evidence for constraints on hawkmoth attraction

Variation in floral phenotype (color, depth, nectar) suggests incipient specialization for bee or hawkmoth pollination across the geographic distribution of Echinopsis ancistrophora, with flower depth ranging from 4.5 to 24 cm. We used chemical and behavioral analyses to test whether fragrance has evolved in concert with morphology in these Andean cacti. Floral scent (145 total compounds) was collected using dynamic headspace methods and analyzed with gas chromatographyÁmass spectrometry, revealing subspecies-specific odors dominated by sesquiterpenes in E. ssp. ancistrophora and arachnacantha and fatty acid derivatives or aromatics in E. ssp. cardenasiana and pojoensis. Compounds indicative of sphingophily were not consistently found in moth-pollinated plants, and total scent emissions were significantly lower in populations with nocturnal anthesis. In wind tunnel assays, Manduca sexta moths were attracted to scent of ssp. ancistrophora from both bee and hawkmoth-pollinated populations, but not to scent of ssp. cardenasiana. However, hawkmoths were most attracted to the methyl benzoate-dominated scent of a distant relative, Echinopsis mirabilis. Thus, hawkmoth-pollinated descendants of the E. ancistrophora lineage may be phylogenetically constrained to emit weak, sesquiterpene-dominated fragrances that are not optimally attractive to hawkmoths, or floral scent may be under stronger selection by destructive flower visitors

Floral scent evolution in the genus jaborosa (Solanaceae): Influence of ecological and environmental factors

Floral scent is a key communication channel between plants and pollinators. However,the contributions of environment and phylogeny to floral scent composition remain poorly understood. In this study, we characterized interspecific variation of floral scent composition in the genus Jaborosa Juss. (Solanaceae) and, using an ecological niche modelling approach (ENM), we assessed the environmental variables that exerted the strongest influence on floral scent variation, taking into account pollination mode and phylogenetic relationships. Our results indicate that two major evolutionary themes have emerged: (i) a ?warm Lowland Subtropical nectar-rewarding clade? with large white hawkmoth pollinated flowers that emit fragrances dominated by oxygenated aromatic or sesquiterpenoid volatiles, and (ii) a ?cool-temperate brood-deceptive clade? of largely fly-pollinated species found at high altitudes (Andes) or latitudes (Patagonian Steppe) that emit foul odors including cresol, indole and sulfuric volatiles. The joint consideration of floral scent profiles, pollination mode, and geoclimatic context helped us to disentangle the factors that shaped floral scent evolution across ?pollinator climates? (geographic differences in pollinator abundance or preference). Our findings suggest that the ability of plants in the genus Jaborosa to colonize newly formed habitats during Andean orogeny was associated with striking transitions in flower scent composition that trigger specific odor-driven behaviors in nocturnal hawkmoths and saprophilous fly pollinators.Fil: More, Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Soteras, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Ibañez, Ana Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Dötterl, Stefan. Paris-lodron-university Of Salzburg; AustriaFil: Cocucci, Andrea Aristides. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Raguso, Robert A.. Cornell University, Dept. Of Neurobiology And Behavior; Estados Unido

Herbivory by a Phloem-Feeding Insect Inhibits Floral Volatile Production

There is extensive knowledge on the effects of insect herbivory on volatile emission from vegetative tissue, but little is known about its impact on floral volatiles. We show that herbivory by phloem-feeding aphids inhibits floral volatile emission in white mustard Sinapis alba measured by gas chromatographic analysis of headspace volatiles. The effect of the Brassica specialist aphid Lipaphis erysimi was stronger than the generalist aphid Myzus persicae and feeding by chewing larvae of the moth Plutella xylostella caused no reduction in floral volatile emission. Field observations showed no effect of L. erysimi-mediated floral volatile emission on the total number of flower visits by pollinators. Olfactory bioassays suggested that although two aphid natural enemies could detect aphid inhibition of floral volatiles, their olfactory orientation to infested plants was not disrupted. This is the first demonstration that phloem-feeding herbivory can affect floral volatile emission, and that the outcome of interaction between herbivory and floral chemistry may differ depending on the herbivore's feeding mode and degree of specialisation. The findings provide new insights into interactions between insect herbivores and plant chemistry

Floral volatiles from Clarkia breweri and C. concinna ( Onagraceae ): Recent evolution of floral scent and moth pollination

Clarkia breweri ( Onagraceae ) is the only species known in its genus to produce strong floral fragrance and to be pollinated by moths. We used gas chromatography-mass spectrometry (GC-MS) to identify 12 abundant compounds in the floral headspace from two inbred lines of C. breweri. These volatiles are derived from two biochemical pathways, one producing acyclic monoterpenes and their oxides, the other leading from phenylalanine to benzoate and its derivatives. Linalool and linalool oxide (pyran form) were the most abundant monoterpenoids, while linalool oxide (furan form) was present at lower concentrations. Of the aromatic compounds detected, benzyl acetate was most abundant, whereas benzyl benzoate, eugenol, methyl salicylate, and vanillin were present as minor constituents in all floral samples. The two inbred C. breweri lines differed for the presence of the additional benzenoid compounds isoeugenol, methyleugenol, methylisoeugenol, and veratraldehyde. We also analyzed floral headspace from C. concinna , the likely progenitor of C. breweri , whose flowers are odorless to the human nose. Ten volatiles (mostly terpenoids) were detected at low concentrations, but only when headspace was collected from 20 or more flowers at a time. Trans-β-ocimene was the most abundant floral compound identified from this species. Our data are consistent with the hypothesized recent evolution of floral scent production and moth pollination in C. breweri.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41639/1/606_2004_Article_BF00983216.pd