Flower movement balances pollinator needs and pollen protection.

Flower signaling and orientation are key characteristics that determine a flower's pollinator guild. However, many flowers actively move during their daily cycle, changing both their detectability and accessibility to pollinators. The flowers of the wild tobacco Nicotiana attenuata orientate their corolla upward at sunset and downward after sunrise. Here, we investigated the effect of different flower orientations on a major pollinator of N. attenuata, the hawkmoth Manduca sexta. We found that although flower orientation influenced the flight altitude of the moth in respect to the flower, it did not alter the moth's final flower choice. These behavioral observations were consistent with the finding that orientation did not systematically change the spatial distribution of floral volatiles, which are major attractants for the moths. Moreover, hawkmoths invested the same amount of time into probing flowers at different orientations, even though they were only able to feed and gather pollen from horizontally and upward-oriented flowers, but not from downward-facing flowers. The orientation of the flower was hence crucial for a successful interaction between N. attenuata and its hawkmoth pollinator. Additionally, we also investigated potential adverse effects of exposing flowers at different orientations to natural daylight levels, finding that anther temperature of upward-oriented flowers was more than 7°C higher than for downward-oriented flowers. This increase in temperature likely caused the significantly reduced germination success that was observed for pollen grains from upward-oriented flowers in comparison to those of downward and horizontally oriented flowers. These results highlight the importance of flower reorientation to balance pollen protection and a successful interaction of the plant with its insect pollinators by maintaining the association between flower volatiles and flower accessibility to the pollinator.

The banana code-natural blend processing in the olfactory circuitry of Drosophila melanogaster.

Odor information is predominantly perceived as complex odor blends. For Drosophila melanogaster one of the most attractive blends is emitted by an over-ripe banana. To analyze how the fly's olfactory system processes natural blends we combined the experimental advantages of gas chromatography and functional imaging (GC-I). In this way, natural banana compounds were presented successively to the fly antenna in close to natural occurring concentrations. This technique allowed us to identify the active odor components, use these compounds as stimuli and measure odor-induced Ca(2+) signals in input and output neurons of the Drosophila antennal lobe (AL), the first olfactory neuropil. We demonstrate that mixture interactions of a natural blend are very rare and occur only at the AL output level resulting in a surprisingly linear blend representation. However, the information regarding single components is strongly modulated by the olfactory circuitry within the AL leading to a higher similarity between the representation of individual components and the banana blend. This observed modulation might tune the olfactory system in a way to distinctively categorize odor components and improve the detection of suitable food sources. Functional GC-I thus enables analysis of virtually any unknown natural odorant blend and its components in their relative occurring concentrations and allows characterization of neuronal responses of complete neural assemblies. This technique can be seen as a valuable complementary method to classical GC/electrophysiology techniques, and will be a highly useful tool in future investigations of insect-insect and insect-plant chemical interactions.

Odour maps in the brain of butterflies with divergent host-plant preferences.

Butterflies are believed to use mainly visual cues when searching for food and oviposition sites despite that their olfactory system is morphologically similar to their nocturnal relatives, the moths. The olfactory ability in butterflies has, however, not been thoroughly investigated. Therefore, we performed the first study of odour representation in the primary olfactory centre, the antennal lobes, of butterflies. Host plant range is highly variable within the butterfly family Nymphalidae, with extreme specialists and wide generalists found even among closely related species. Here we measured odour evoked Ca(2+) activity in the antennal lobes of two nymphalid species with diverging host plant preferences, the specialist Aglais urticae and the generalist Polygonia c-album. The butterflies responded with stimulus-specific combinations of activated glomeruli to single plant-related compounds and to extracts of host and non-host plants. In general, responses were similar between the species. However, the specialist A. urticae responded more specifically to its preferred host plant, stinging nettle, than P. c-album. In addition, we found a species-specific difference both in correlation between responses to two common green leaf volatiles and the sensitivity to these compounds. Our results indicate that these butterflies have the ability to detect and to discriminate between different plant-related odorants.

Time and space are complementary encoding dimensions in the moth antennal lobe.

The contribution of time to the encoding of information by the nervous system is still controversial. The olfactory system is one of the standard preparations where this issue is empirically investigated. For instance, output neurons of the antennal lobe or the olfactory bulb display odor stimulus induced temporal modulations of their firing rate at a scale of hundreds of milliseconds. The role of these temporal patterns in the encoding of odor stimuli, however, is not yet known. Here, we use optical imaging of the projection neurons of the moth antennal lobe to address this question. First, we present a biophysically derived model that provides an accurate description of the calcium response of projection neurons. On the basis of this model, we subsequently show that the calcium response of the projection neurons displays a stimulus specific temporal structure. Finally, we demonstrate that an encoding scheme that includes this temporal information boosts classification performance by 60% as compared to a purely spatial encoding. Although the putative role of combinatorial spatio-temporal encoding strategies has been the subject of debate, our results for the first time establish quantitatively that such an encoding strategy is used by the insect brain.

Attractiveness of fruit and flower odorants detected by olfactory receptor neurons in the fruit chafer Pachnoda marginata.

We studied the attraction of the African fruit chafer Pachnoda marginata Drury (Coleoptera: Scarabaeidae) to banana and 34 synthetic plant compounds previously shown to be detected by P. marginata olfactory receptor neurons. The behavioral studies were carried out in a two-choice olfactometer, where the attraction of beetles to lures and controls was monitored in 30-min intervals during whole days. Monitoring of the attraction over time gave additional information when comparing relative attractiveness of different compounds. Seventeen of the test compounds, primarily phenylic compounds, fruit esters, isovaleric acid, acetoin, and some floral or fruit terpenes, were attractive to P. marginata. Compounds showing no attractiveness included green leaf volatiles, lactones. and several alcohols, but also phenylic compounds and esters. One case of blend synergism was demonstrated, as well as some examples of sexual dimorphism in attraction. The significance of certain compounds and receptor neurons for olfactory-guided behavior of phytophagous scarabs is discussed.

Novel natural ligands for Drosophila olfactory receptor neurones.

Due to its well-defined genome, the fruitfly Drosophila melanogaster has become a very important model organism in olfactory research. Despite all the research invested, few natural odour ligands have been identified. By using a combined gas chromatographic-single receptor neurone recording technique (GC-SC), we set out to identify active odour molecules in head space-collected volatiles from preferred food sources, i.e. different overripe or rotting fruit. In total, we performed 101 GC-SC experiments on 85 contacted sensilla. Using GC-mass spectrometry, we identified 24 active compounds. Synthetic samples of these compounds were used to establish dose-response curves for several of the receptor neurone types encountered. The response patterns of individual neurones were repeatable, and neurones were found to reside in stereotyped pairs. In total, we identified eight distinct sensillum types based on response profiles of 12 olfactory receptor neurone types. In most recordings, a single GC peak would produce a strong response, whereas a few other, often chemically related, compounds would produce weaker responses. The GC-SC recordings revealed that the olfactory receptor neurones investigated were often selective and could be divided into distinct functional types with discrete characteristics. Dose-response investigations revealed very low response thresholds to the tested compounds. Six of the novel ligands were also tested for their behavioural effect in a T-maze set up. Of these, five elicited attraction and one elicited repulsion.