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Background and Aims The first documented observation of pollination in Pleurothallidinae was that of Endre ́s, who noticed that the ‘viscid sepals’ of Specklinia endotrachys were visited by a ‘small fly’. Chase would later iden- tify the visiting flies as being members of the genus Drosophila. This study documents and describes how species of the S. endotrachys complex are pollinated by different Drosophila species. Methods Specimens of Specklinia and Drosophila were collected in the field in Costa Rica and preserved in the JBL and L herbaria. Flies were photographed, filmed and observed for several days during a 2-year period and were identified by a combination of non-invasive DNA barcoding and anatomical surveys. Tissue samples of the sepals, petals and labellum of Specklinia species were observed and documented by SEM, LM and TEM. Electroantennogram experiments were carried out on Drosophila hydei using the known aggregation pheromones ethyl tiglate, methyl tiglate and isopropyl tiglate. Floral compounds were analysed by gas chromatography–mass spectometry using those same pheromones as standards. Key Results Flowers of S. endotrachys, S. pfavii, S. remotiflora and S. spectabilis are visited and pollinated by several different but closely related Drosophila species. The flies are arrested by aggregation pheromones, including ethyl tiglate, methyl tiglate and isopropyl tiglate, released by the flowers, and to which at least D. hydei is very sen- sitive. Visible nectar drops on the adaxial surface of sepals are secreted by nectar-secreting stomata, encouraging male and female Drosophila to linger on the flowers for several hours at a time. The flies frequently show courtship behaviour, occasionally copulating. Several different Drosophila species can be found on a single Specklinia species. Conclusions Species of the S. endotrachys group share a similar pollination syndrome. There seem to be no spe- cies-specific relationships between the orchids and the flies. It is not expected that Specklinia species will hybridize naturally as their populations do not overlap geographically. The combination of pheromone attraction and nectar feeding is likely to be a generalized pollination syndrome in Pleurothallidinae. Ministerio del Ambiente y Energía/[]/MINAE/Costa Rica Sistema Nacional de Áreas de Conservación/[]/SINAC/Costa Rica Universidad de Costa Rica/[814-B1-239]/UCR/Costa Rica Universidad de Costa Rica/[814-B3-075]/UCR/Costa Rica UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Jardín Botánico Lankester (JBL)

Anemotaxis in the predatory mite Phytoseiulus persimilis (both well-fed and starved), has previously been studied on a wire grid under slight turbulent airflow conditions yielding weak, yet distinct, gradients in wind speed and odour concentration (Sabelis and Van der Weel 1993). Such conditions might have critically influenced the outcome of the study. We repeated these experiments, under laminar airflow conditions on a flat surface in a wind tunnel, thereby avoiding variation in wind speed and odour concentration. Treatments for starved and well-fed mites were (1) still-air without herbivore-induced plant volatiles (HIPV) (well-fed mites only), (2) an HIPV-free air stream, and (3) an air stream with HIPV (originating from Lima bean plants infested by two-spotted spider mites, Tetranychus urticae). Well-fed mites oriented in random directions in still-air without HIPV. In an air stream, starved mites always oriented upwind, whether plant odours were present or not. Well-fed mites oriented downwind in an HIPV-free air stream, but in random directions in an air stream with HIPV. Only under the last treatment our results differed from those of Sabelis and Van der Weel (1993).

Two compounds present on the surface of Brassica oleracea cv. botrytis leaves have been isolated and identified which stimulate very effectively oviposition in the cabbage root fly, Delia radicum and which are perceived by a specific receptor neuron in the tarsal sensillum C5 of the female fly. Activity of extracts and chromatographic fractions were bioassayed, using oviposition experiments and mainly electrophysiological recordings from the C5 tarsal contact chemoreceptor sensillum of female flies. Spectroscopic data indicate that the main compound is 1,2-dihydro-3-thia-4,10,10b-triaza-cyclopenta[.a.]fluorene-1-carboxylic acid, a novel compound related to Brassica phytoalexins like brassicanal C. It is accompanied by its glycine conjugate.

Summary Small ermine moths (Yponomeutidae: Lepidoptera) are specialist herbivores. Species within the genus Yponomeuta are each specialized on a limited number of plant species, mainly within genera belonging to the Celastraceae. European Yponomeuta species have developed new specialized host affiliations, mainly on rosaceous hosts. Since these host shifts are reputed to be of consequence for speciation, the role of the ovipositing female is of particular interest. Study of the pre-oviposition behaviour of gravid Y. cagnagellus (Hb.) moths on host (Euonymus europaeus), non-host (Crateagus monogyna) and artificial oviposition substrates, provided information on the nature of the cues used for host plant acceptance and the insect’s perception of these cues. Host selection by adult females occurs with contact chemoreceptors probably located on the antennae or tarsi. MeOH-soluble, non-volatile phytochemical compounds washed from the host plant’s surface and applied on an artificial twig are sufficient to stimulate a complete sequence of behavioural elements leading to oviposition. Volatiles do not have a large effect on the pre-oviposition behaviour.

Yponomeuta cagnagellus is a phytophagous moth species specialized on Euonymus europaeus. Host discrimination by the adult female is an important aspect of host specialization and is based mainly on the distinctive secondary chemistry of host and nonhosts. This paper describes a bioassay that was developed to study the effect of isolated plant surface compounds on Yponomeuta oviposition. Adult moths recognize their hosts through chemical stimuli on the leaf or twig surface. Relatively apolar compounds extracted from the host twig surface by washing in dichlormethane do not stimulate oviposition. More polar, methanol-soluble compounds do, and this stimulation is dose dependent. Moths are able to recognize hosts solely by their surface compounds: females show a strong preference for artificial twigs treated with methanolic extracts of their hosts compared to those treated with methanolic extracts of nonhosts Crataegus monogyna and Prunus spinosa (both of which are hosts for closely related Y. padellus). Shape and surface characteristics of the oviposition substrate also influence oviposition. The substrate needs to resemble the basic form of a twig (i.e., cylindrical), and females prefer a coarse surface with irregularities over a smooth one.

The effects of six terpenoids and two terpenoid containing extracts of the neem tree (Azadirachta indica A. Juss.) on oviposition by the diamondback moth (Plutella xylostella L., Yponomeutidae: Lepidoptera) were tested. Two drimane terpenoids, the sesquiterpenoid polygodial and the neem extract Margosan-O exerted significant inhibitory effects at the dosages tested. Ablation experiments showed that both antennae and fore-tarsi contributed to mediation of the inhibition by a drimane. Location of chemosensilla on prothoracic tarsi and ovipositor was examined by scanning electronmicroscopy. Electrophysiological recordings from ovipositor and tarsal taste sensilla showed that distilled water produced distinct responses from one neuron. In tarsal sensilla, ethanol and drimane solutions produced responses from two neurons, one of which might be the water cell that fired at a reduced rate. A drimane significantly decreased the responses of tarsal chemoreceptors to a cabbage leaf extract, which is a possible sensory mechanism leading to behavioural avoidance of this compound.

Contact chemoreception plays a decisive role in host selection and oviposition behaviour of the cabbage root fly, Delia radicum L. (Diptera, Anthomyiidae). Glucosinolates (mustard oil glucosides) are known to be perceived by the flies, and when sprayed on paper leaf-models induce oviposition. Recently it has become clear that other non-volatile types of compounds must also be involved in host selection. A pair of ventro-medial C sensilla on die fifth tarsomere respond strongly to a novel compound called tentatively ‘cabbage identification factor’ (CIF), but not to sucrose, glucose, fructose and proline. CDF is a new non-glucosinolate oviposition stimulant. A single neurone in each sensillum is activated by this compound and the same is true for glucosinolates. In some flies a mixture of bom types of stimuli evoked an apparent mononeural spike train, whereas in odiers spikes of two separate cells were activated. The significance of this variability is not yet clear. The new stimulant, CIF, does not evoke responses in glucosinolate receptors in the D sensilla. The involvement of the C3 sensilla in the detection of host-specific compounds constitutes the first known function for C sensilla in D. radicum. CIF appears to be present in leaf surface extracts from the host-plant Brassica oleracea in quantities as low as 1 ng per gram leaf. In spite of this low level, it stimulates oviposition significantly better than glucobrassicin at higher concentrations, which up till now was known as the most powerful stimulant for D.radicum.

The role of glucosinolates in the oviposition behaviour of the cabbage root fly, Delia radicum (L.) (Diptera, Anthomyiidae) was investigated using egg counts and electrophysiological recordings from tarsal contact chemoreceptors. The glucosinolates present both inside and on the surface of cauliflower leaves were determined. The total amounts obtained with the two methods differed by a factor of 100. The extract of the leaf surface contained about 60 μg per g leaf extracted (gle), the total leaf extract 7.5 mg per gle. The glucosinolate patterns of the two extracts were qualitatively similar, but the ratios of the content of individual glucosinolates showed considerable differences. The D sensilla on segment 3 and 4 of the tarsus of D. radicum females were shown to contain a sensitive receptor cell for glucosinolates. In contrast, the receptor cells of the D sensilla of the other segments did not respond in a dose dependent way to these compounds. The glucosinolate receptors were found to be especially sensitive to glucobrassicin, gluconasturtiin and glucobrassicanapin with thresholds of about 10 −8M to 10 −9M. Large differences (up to two orders of magnitude) were observed among the different glucosinolates. A significant correlation was found between the behavioural discrimination index and the electrophysiological results. But no obvious correlation existed between the chemical nature of the glucosinolate side chain (e.g. indole, aromatic and aliphatic groups), and their stimulatory activity. However, a significant correlation was found between the overall length of the side chain and the biological activity. Although the flies discriminated clearly between model leaves with and without glucosinolates, a clear dose response curve was only obtained for the indole glucosinolate glucobrassicin. Since the most stimulatory fraction of the surface extract contained no glucosinolates, it was concluded that other compounds, in addition to glucosinolates, do play an important role for the stimulation of oviposition.

Tarsal contact chemoreceptors of the black swallowtail butterfly Papilio polyxenes (Lepidoptera: Papilionidae) were stimulated with leaf-surface extracts and ethanolic extracts of whole leaves of a host-plant (Daucus carota) and a non-host (Brassica oleracea). Both leaf extracts evoked large numbers of spikes but stimulated different receptor neurones. It is concluded that a large difference exists between the sensory responses to host and non-host extracts. Two chemicals, luteolin 7-0-(6“-0-malonyl)-β-D-glucopyranoside and trans-chlorogenic acid, known to be present in the host and known oviposi-tion stimulants for P.polyxenes, were also tested and shown to be active. No responses were found to luteolin 7-O-β-D-glucoside or to luteolin 7-O-β-D-glucuronide. These flavonoids occur in D.carota foliage, but do not stimulate oviposition.

The effects of foliar form, colour and surface characteristics on the oviposition behaviour of the cabbage fly were investigated by presenting females with various leaf models made of paper. The models differed in shape, size, colour, surface coating, the presence of 3 dimensional folds and a stem. It was shown that in the presence of host leaf surface extracts physical factors can strongly influence oviposition behaviour. Females laid the most eggs around the base of bright green or yellow models having a stem, vertical folds and covered with a thin layer of paraffin. The vertical folds in the surface increased the chance of a behavioural transition from leaf run to stem run. Zusammenfassung Eiablageverhalten der Kohlfliege beeinflusst durch Blatt-Form, Farbe und Oberflachestructur Mit Hilfe von Papiermodellen von Kohlblattern wurde der Einfluss von visuellen und mechanischen Eigenschaften des Blattes auf das Eiablageverhalten der kleinen Kohlfliege, Delia radicum, untersucht. Die verwendeten Modelle unterschieden sich in Form, Grosse, Farbe, Oberflachen struktur und Stiel des Blattes. Es wurde gezeigt, dass physikalische Reize zusammen mit den chemischen Stoffe auf der Blattoberflache stimulierend wirken. Ein naturliches Eiablageverhalten und die starkste Eiablage wurde durch ein Modell ausgelost, welches sich durch leuchtend grune Farbe, Stiel und vertikale Falten und einen Paraffinnuberzug auszeichnete.

In lepidopterous larvae the maxillary palps contain a large portion of the sensory equipment of the insect. Yet, knowledge about the sensitivity of these cells is limited. In this paper a morphological, behavioral, and electrophysiological investigation of the maxillary palps of Yponomeuta cagnagellus (Lepidoptera: Yponomeutidae) is presented. In addition to thermoreceptors, CO(2) receptors, and gustatory receptors, evidence is reported for the existence of two groups of receptor cells sensitive to plant volatiles. Cells that are mainly sensitive to (E)-2-hexenal and hexanal or to (Z)-3-hexen-1-ol and 1-hexanol were found. Interestingly, a high sensitivity for benzaldehyde was also found. This compound is not known to be present in Euonymus europaeus, the host plant of the monophagous Yponomeuta cagnagellus, but it is a prominent compound in Rosaceae, the presumed hosts of the ancestors of Y. cagnagellus. To elucidate the evolutionary history of this sensitivity, and its possible role in host shifts, feeding responses of three Yponomeuta species to benzaldehyde were investigated. The results confirm the hypothesis that the sensitivity to benzaldehyde evolved during the ancestral shift from Celastraceae to Rosaceae and can be considered an evolutionary relict, retained in the recently backshifted Celastraceae-specialist Y. cagnagellus.

Changes in host acceptance is an important factor in the host specialization of phytophagous insects, and knowledge of the genetic organization of this behaviour is necessary in order to understand how host shifts occur. Here we describe the inheritance of adult host acceptance (oviposition) in three closely related species of Yponomeuta Latreille (Lepidoptera: Yponomeutidae), and their interspecific hybrids. Yponomeuta cagnagellus (Hubner), a specialist on Euonymus europaeus L. (Celastraceae), Y. malinellus Zeller, a specialist on Malus spp. (Rosaceae), and Y. padellus (L.), oligophagous on a number of Rosaceae, were tested for their acceptance of parental hosts in choice tests. Acceptance of E. europaeus is semi-dominant in hybrids of Y. cagnagellus x Y. padellus, and in hybrids of Y. cagnagellus x Y. malinellus. The dominance of this acceptance was confirmed in oviposition tests with backcross hybrids: backcross hybrids Fl x Y. cagnagellus oviposited mainly on E. europaeus and Fl x Y. padellus still deposited more than half of their egg masses on E. europaeus. Reciprocal hybrids did not differ in their host acceptance, indicating that the trait is autosomal. We further studied the effect of larval food on adult host acceptance ('Hopkins host selection principle') in split full-sib F1 families. Larval diet influenced oviposition only in one of two hybrid crosses. The F1 hybrid of Y. padellus x Y. cagnagellus, reared on Prunus spinosa L., deposited a significantly lower percentage of egg masses on E. europaeus compared to their full-sib sisters fed with E. europaeus. We did not find this in the reciprocal cross. However, still more than half of the egg masses are deposited on E. europaeus by hybrids that have no experience on this host. We conclude that the semi-dominant character of acceptance of E. europaeus and a tendency of Rosaceae-feeding Yponomeuta to deposit egg masses on this host may have created the opportunity for the host shift of the predecessor of Y. cagnagellus from Rosaceae to the Celastraceae. This shift may have been further facilitated by a weak tendency of adults to oviposit on their larval food source.

The cellular localisation of the sensitivity to two host-plant specific larval phagostimulants was determined in Yponomeuta evonymellus and in inter-specific hybrids of Y. cagnagellus and Y. padellus. The combined results of cross-adaptation experiments, analysis of mixture responses and inter-spike interval distributions indicated that the sensitivity for sorbitol and dulcitol is localised in the same cell in the investigated species. This result suggests that during the evolution of the genus the main alteration in the mechanism of sugar-alcohol detection occurred at the level of the receptor proteins. Relatively simple modification of this kind can, however, have large effects, since larvae with modified receptor proteins will directly respond to the new stimulus while all central processing and resulting behaviour can remain unmodified.

The behavioural phase state and coloration of hatchling Schistocerca gregaria were examined in a series of experiments to determine the means by which phase characteristics are passed between generations. Both crowding of solitary-reared females at the time of oviposition and high egg pod densities promoted behavioural gregarization, although the former appeared to be a rather more potent factor. In contrast, egg pod density alone appeared to promote the development of hatchlings with dark patterns characteristic of the gregarious phase. The phase characteristics of hatchlings were unaffected when sand previously used for oviposition was used to collect further egg pods. Early separation of presumptive gregarious eggs from egg pods laid by crowd-reared females led to solitarization of the hatchlings, indicating that a factor, either in or around the eggs, removed by early separation promoted gregarization. Both the eggs and foam plugs of egg pods from crowd-reared, gregarious females appeared to be a source of this gregarizing factor. In contrast, there was no evidence for a solitarization factor in egg pods from solitary-reared S. gregaria. Saline extracts of egg pod foam plugs produced an active factor which promoted gregarization both in eggs from solitary-reared females and in eggs from gregarious females which were separated and washed to removed the factor. Solitary eggs were influenced by the gregarizing factor in foam plug extracts for up to 1 day after oviposition. Saline extracts of foam plug retained their activity for up to 1 day. Initial studies on the properties of this factor were made. We conclude that the foam plugs of egg pods from crowd-reared, gregarious locusts contain a small (

Oviposition by the cabbage root fly, Delia radicum (L.) (Dipt., Anthomyiidae) is governed to a large extent by chemicals present on the leaf surface of the host plant Brassica oleracea L. Although this fact has been recognized for some time, knowledge about the nature of the phytochemicals is still far from complete. Glucosinolates (mustard oil glucosides) and their breakdown products, the isothiocyanates (mustard oils), have been known to be involved. The glucosinolates stimulate oviposition and the tarsal D-sensilla contain chemoreceptors sensitive to this group of compounds (Stadler, 1978; Roessingh et al., unpubl.).