Larval chemical defence and evolution of host shifts in Chrysomela leaf beetles

Summary. Larvae of Chrysomela leaf beetles release for defence volatile compounds belonging to various chemical families. This study focuses on the defensive strategy based on the esterification of isobutyric acid and 2-methylbutyric acid with a wide variety of alcohols taken up from the host plant. To date, only two species are known to produce these repellents C. interrupta, which is associated with Betulaceae and C. lapponica which occurs either on Betulaceae or Salicaceae.? In order to know if other species have developed this chemical defence and how the food plant influences the secretion of these toxins, we targeted by mass spectrometry the presence of iso- and 2-methylbutyric acids and esters of them in the defensive secretions of Chrysomela larvae exclusively associated with Betulaceae or Salicaceae. ?Screening analyses reveal that the synthesis of these compounds is a common character restricted to all the members belonging to the C. interrupta group sensu Brown (1956) regardless of the host-plant family. These results suggest that the biochemical mechanism leading to the synthesis of these compounds could be considered as a synapomorphy meaning that the group is probably monophyletic. ?Defensive secretions of the members of the interrupta group are quantitatively assayed for iso- and 2-methylbutyric acids and their (Z)-3-hexenyl esters. Results reveal a chemical plasticity developed by Chrysomela species associated with Salicaceae. The amounts of iso- and 2-methylbutyric acids derivatives and of salicylaldehyde in their larval secretions depend on the food plant and on its content in phenolglucosides. Received 5 October 1998; accepted 25 November 1998.     

Waterborne chemical compounds in tropical macroalgae: positive and negative cues for larval settlement

Settlement sites of marine invertebrate larvae are frequently influenced by positive or negative cues, many of which are chemical in nature. Following from the observation that many shallow-water, Hawai'ian marine macroalgae are free of fouling by sessile invertebrates, we predicted that the algae are chemically protected and dependent on either surface-bound or continuously released soluble compounds to deter settling invertebrate larvae. To address the importance of waterborne algal compounds, we experimentally determined whether larvae of two of Hawai'i's dominant hard-surface fouling organisms, the polychaete tube worm Hydroides elegans and the bryozoan Bugula neritina, would settle in the presence of waters conditioned by 12 species of common Hawai'ian macroalgae (representing the Phaeophyta, Chlorophyta, Rhodophyta and Cyanophyta). The results included a full spectrum of biological responses by each larval species to waterborne algal compounds. Larval responses to conditioned water were consistent for each algal species, but the outcomes were not predictable based on the taxonomic relationships of the algae. For example, among the species of Phaeophyta examined, different conditioned waters were: (1) toxic, (2) inhibited settlement, (3) simulated settlement, or (4) had no effect, compared to larvae in control dishes containing filtered seawater. Additionally, larval responses to aged (24 h) conditioned waters could not be predicted from the results of assays run with conditioned waters utilized immediately after preparation. Finally, settlement by larvae of one species did not predict outcomes of tests for the other species. Four of 12 shallow-reef Hawai'ian macroalgae tested released compounds into surrounding waters that immediately killed or inhibited settlement by both H. elegans and B. neritina (toxic: Dictyota sandvicensis; inhibitory: Halimeda discoidea, Sphacelaria tribuloides, Ulva reticulata); the remaining 8 algal species prevented settlement by one of these fouling organisms but for the other had no effect or, in some cases, even stimulated settlement     

Role of surface chemical signals in egg cannibalism and intraguild predation in ladybirds (Coleoptera: Coccinellidae)

Summary. The eggs of some ladybirds are known to be toxic to intraguild ladybird predators. However, this defence is of little value if the eggs are killed before their toxicity becomes apparent. The results presented in this paper indicate that chemicals on the surface of the eggs of two species of ladybirds signal the relative risk of cannibalism and intraguild predation. In Adalia bipunctata and Coccinella septempunctata, 87% of the chemicals are alkanes. Each species of ladybird is less reluctant to eat their own eggs than those of the other species. This asymmetry is to be expected because there is a greater risk to ladybirds from intraguild predation than cannibalism. Similar alkanes to those on the surface of the eggs of A. bipunctata are present in tracks left by larvae and on the elytra of the adults of this species. Those in the larval tracks deter females from ovipositing in patches of prey already being attacked by their larvae and those on the elytra are used in mate recognition. That different context dependent messages could be signalled by similar chemicals is an example of semiochemical parsimony. Received 25 February 2000; accepted 22 May 2000     

Chemical mimicry in the root aphid parasitoid Paralipsis eikoae Yasumatsu (Hymenoptera: Aphidiidae) of the aphid-attending ant Lasius sakagamii Yamauchi & Hayashida (Hymenoptera: Formicidae)

Summary. An aphidiid wasp, Paralipsis eikoae, was associated with both Lasius niger and L. sakagamii attending the wormwood root aphid Sappaphis piri. An L. sakagamii worker was observed carrying a winged female P. eikoae to its nest with its mandible, but it did not kill the wasp. Once accepted by the ants, the wasp often mounted and rubbed against the worker ants and sometimes teased them to regurgitate food to itself. No workers in the colony attacked the wasp. Conspecific foreign workers, however, viciously attacked the wasp when encountered. Gas chromatography-mass spectrometry analyses showed that the accepted wasp had complex cuticular hydrocarbons that were very similar to those of its host ants, whereas the winged wasps collected outside the ant nest showed only a series of n-alkanes. Additionally, the accepted wasp had a hydrocarbon profile closer to that of its host ants than to the conspecific foreign ants. We believe the wasp mimics ant cuticular hydrocarbons to integrate into the ant nest, acquiring the hydrocarbons by mounting and rubbing against the ants. In contrast, the cuticular hydrocarbons of the emerged wasp contained larval and pupal hydrocarbons of L. sakagamii that were also similar to those of L. niger. Both ant species rejected adult workers of the other species but accepted their larvae and pupae. We suggest that the emerged P. eikoae mimics the cuticular hydrocarbons of these Lasius larvae and pupae, which allows P. eikoae to be accepted by both L. sakagamii and L. niger. Received 11 March 1998; accepted 22 July 1998.     

Cuticular volatiles,attractivity of worker larvae and invasion of brood cells by Varroa mites. A comparison of Africanized and European honey bees

Summary. Africanized honey bees (AHBs) of Brazil and Mexico have proven to be tolerant to Varroa destructor mites. In contrast, European honey bees (EHBs: Apis mellifera carnica) at the same tropical study site are highly intolerant to these ectoparasites. A lower attractiveness of Varroa-tolerant AHB larvae has been hypothesised to be an important trait in reducing the susceptibitlity of AHBs to these mites. Thus, selection for EHB brood that is less attractive to mites is thought to be one possibility for limiting mite population growth and thus increase the tolerance of EHBs to the mite.?In Ribeir?o Preto, Brazil, European A. m. carnica bees and AHBs were tested with respect to their rate of brood infestation and brood attractiveness to Varroa mites. For the comparison of brood infestation rates, we introduced combs with pieces of EHB and AHB brood into honey bee colonies (18 repetitions). The relative infestation rate of EHB brood was significantly higher compared to AHB brood.?The preference behaviour of single Varroa mites was tested in a laboratory bioassay where either living host stages were offered or host extracts were presented on dummies. By these tests we could confirm the preference of Varroa females for certain developmental host stages and for their corresponding extracts. In contrast to the within-colony results, Varroa mites in the laboratory bioassay showed a slight preference for AHB compared to EHB larvae.?The gas chromatographic analysis revealed differences in the chemical spectrum of extracts obtained from different larvae. In accord with the results of the bioassays, we could detect stage-specific odour differences in larval cuticular compounds, including methyl esters and hydrocarbons that have been described as kairomones. None of these substances, however, revealed significant race-specific differences. Therefore, the quantity and composition of certain cuticular compounds seem to be responsible only for the recognition of a suitable host stage by Varroa females. The different infestation rates in the colonies, however, seem to be caused neither by race-specific differences in attractiveness of bee larvae nor by an extended attractive period of EHB larvae: both AHB and EHB larvae become attractive approximately 21 h before capping of the brood cell, and thus have the same window of time when they can be parasitised.?Therefore differential Varroa-infestation rates are not related to larval attraction but probably are determined by other race-specific and colony-related factors. Received 11 June 2001; accepted 19 November 2001.     

Damage-inducibility of primary and secondary metabolites in the wild parsnip (Pastinaca sativa)

Summary. Induction of secondary metabolites to herbivore damage is a widespread phenomenon among plants and serves to enhance resistance by reducing suitability or increasing toxicity of foliage. Post-damage responses of primary metabolites are less well known; reductions in primary metabolites may increase resistance by decreasing palatability or nutritional suitability for herbivores or by potentiating toxicity of secondary metabolites. In this study, we examined response to simulated herbivory in Pastinaca sativa, the wild parsnip, in both primary and secondary metabolites. We found that induction of secondary metabolites in response to damage is largely restricted to a single class of compounds, the furanocoumarins. These prooxidant compounds are known to be toxic to a wide variety of insect herbivores. The only primary metabolite that responded to damage was total soluble protein, which increased significantly 24 h after damage. After 24 h, the correlation between total furanocoumarins and total sugars was significant and negative (r = − 0.697). This correlation likely reflects an independent response of sugar to damage, rather than a diversion of resources into furanocoumarin production, inasmuch as this correlation at 3 h, after furanocoumarin induction had taken place, was not significant. In other secondary metabolite pathways, damage produced a significant decline in farnesene and a C-16 unsaturated fatty acid, 7,10,13-hexadecatrienoic acid, each of which may potentiate the furanocoumarin defense response. Farnesene may enhance photooxidative activation of the furanocoumarins and 7,10,13-hexadecatrienoic acid may serve as a precursor to jasmonic acid, a key hormone in regulating induction responses. With few key exceptions, quantities of both primary and secondary metabolites in wild parsnip foliage are unaffected by damage. Those that are affected may well play a role in resistance of parsnips to herbivores. Received 1 July 1998; accepted 28 September 1998.     

Lack of sequestration of host plant glucosinolates in <Emphasis Type="Italic">Pieris rapae</Emphasis> and <Emphasis Type="Italic">P. grarricae</Emphasis>

Summary. Sequestration of plant toxins in herbivores is often correlated with aposematic coloration and gregarious behaviour. Larvae of Pieris brassicae show these conspicuous morphological and behavioural characteristics and were thus suggested to sequester glucosinolates that are characteristic secondary metabolites of their host plants. P. rapaeare camouflaged and solitary, and are thus not expected to sequester. To test this hypothesis and to check the repeatabi-lity of a study that did report the presence of the glucosinolate sinigrin in P. brassicae, larvae were reared on three species of Brassicaceae (Sinapis alba, Brassica nigra and Barbarea stricta), and different leaf and insect samples were taken for glucosinolate analysis. The major host plant glucosinolates could only be found in traces or not at all in larval haemolymph, bled or starved larvae, faeces or pupae of both species or P. brassicae regurgitant. Haemolymph of both Pieris spp. was not rejected by the ant Myrmica rubra in dual-choice assays; the regurgitant of P. brassicae was rejected. This suggests the presence of compounds other than glucosinolates that might be sequestered in or produced by P. brassicae only. In faeces of both Pieris spp. a compound which yielded 4-hydroxybenzylcyanide (HBC) upon incubation with sulfatase was detected in high concentrations when larvae had been reared on S. alba. This compound may be derived from hydrolysis of sinalbin, the main glucosinolate of that plant. The unidentified HBC progenitor was apparently not sequestered in the two Pieris spp., and was not detected in faeces of larvae reared on B. nigra or B. stricta. Received 18 July 2002; accepted 11 September 2002.     

Sequestration of aristolochic acids by the pipevine swallowtail, Battus philenor (L.): evidence and ecological implications

Summary. It has long been assumed that the North American pipevine swallowtail, Battus philenor (L.) (Papilionidae, Troidini), is protected from natural enemies by aristolochic acids sequestered from its Aristolochia food plants. This study confirmed that populations of B. philenor from Virginia and east Texas sequester these compounds. A comparison of the aristolochic acid profiles of the Virginia butterflies and their A. macrophylla food plants revealed several differences. The aristolochic acid fraction of the foliage was dominated by aristolochic acids I and II, whereas the insects had a much lower proportion of aristolochic acid II and contained, in addition, substantial amounts of aristolochic acids Ia and IVa, which were not detected in the plants. The eggs, larval integument, osmeterial glands, pupal cuticle, and adults (wings and bodies) all contained aristolochic acids. These findings help explain the abundant ecological data indicating that both immature and adult B. philenor are unpalatable and protected from natural enemies. Received 7 April 2000; accepted 31 May 2000     

Comparative studies on the sex pheromones of Ostrinia spp. in Japan: the burdock borer, Ostrinia zealis

Summary. To gain insight into the evolution of the sex pheromone communication system in Ostrinia (Lepidoptera Pyralidae), the sex pheromone of the burdock borer, O. zealis was analyzed by means of gas chromatography-electroantennographic detection (GC-EAD), GC-mass spectrometry and a series of bioassays. Four EAD-active compounds were detected in the female sex pheromone gland extract, and these were identified as tetradecyl acetate (14:OAc), (Z)-9-tetradecenyl acetate (Z9–14:OAc), (E)-11-tetradecenyl acetate (E11-14:OAc) and (Z)-11-tetradecenyl acetate (Z11-14:OAc). The average amounts (ratio) of the four compounds in single sex pheromone glands were 2.5 ng (13%), 11.6 ng (61%), 4.1 ng (21%) and 0.9 ng (5%), respectively. In a wind-tunnel bioassay, the ternary blend of Z9-, E11- and Z11-14:OAc at a ratio found in the sex pheromone gland elicited the same behavioral responses from the males as did virgin females. 14:OAc did not show any enhancement or inhibition of the males’ behavioral responses when added to the ternary blend. The attractiveness of the 3-component lure to O. zealis males was also confirmed by field trapping experiments. Based on these results, we concluded that the sex pheromone of O. zealis is composed of Z9-14:OAc, E11-14:OAc and Z11-14:OAc at a ratio of 70:24:6. The evolutionary changes of the sex pheromones in Ostrinia are also discussed based on the presently available information on the sex pheromones and phylogenetic relationships of Ostrinia spp. Received 25 September 1998; accepted 2 December 1998.     

A trail pheromone component of the African stink ant, Pachycondyla (Paltothyreus) tarsata Fabricius (Hymenoptera: Formicidae: Ponerinae)

Summary. The African stink ants (Pachycondyla tarsata) lay recruitment trails with secretions from sternal glands. The glandular secretions consist of 10 compounds, 9 of which have been chemically identified. One of the substances, 9-heptadecanone, elicits trail following behavior in P. tarsata workers that have before been stimulated by a sucessful scout ant. Received 7 August 1998; accepted 24 November 1998.     

The occurrence of aristolochic acids in neotropical troidine swallowtails (Lepidoptera: Papilionidae)

Summary. Analysis of individuals of 17 troidine species demonstrated the presence of aristolochic acids in these butterflies in varying concentrations. Although aristolochic acids do not occur in Aristolochia galeata leaves, they were present in Battus polydamas larvae reared on these leaves, and thus may be synthesized by the larvae from chemical precursors in the plant. Received 17 August 1999; accepted 20 January 2000     

Sequestration of dietary secondary metabolites by three species of sea hares: location,specificity and dynamics

We examined the location of squuestered secondary metabolites in three species of sea hares, Stylocheilus longicauda, Dolabella auricularia, and Aplysia californica (Opisthobranchia: Anaspidea). The sea hares ate a natural diet or were fed an artificial diet containing secondary metabolites in the laboratory. In all three species, sequestered secondary metabolites were located almost exclusively in the digestive gland, an internal organ, rather than in the exterior parts of the body, in eggs, or in ink (released when sea hares are disturbed). S. longicauda, a specialist sea hare, was able to sequester measur-able amounts of all six algal metabolites offered (caulerpenyne, halimedatetraacetate, pachydictyol A, malyingamides A and B, and ochtodene) and two (luffariellolide and Dysidea spp. brominated diphenyl ether) of three sponge metabolites offered (chondrillin was not sequestered). Malyngamides A and B, found in the host plant of S. longicauda, were sequestered at high, but not unique concentrations. D. auricularia, a generalist sea hare, was fed caulerpenyne, pachydictyol A and malyngamide B; patterns of sequestration of these three compounds did not differ markedly between S. longicauda and D. auricularia. S. longicauda did not lose measurable amounts of malyngamides after 18 d on a malyngamide-free diet. These results suggest that sea hares have generic mechanisms for sequestering algal metabolites rather than mechanisms that are tightly linked to particular compounds, that these mechanisms do not differ dramatically between species, and that sequestered secondary metabolites are not located optimally for defense.     

Dynamics of pheromone production and communication in the mountain pine beetle, Dendroctonus ponderosae Hopkins, and the pine engraver, Ips pini (Say) (Coleoptera: Scolytidae)

Summary. The mountain pine beetle, Dendroctonus ponderosae Hopkins, and the pine engraver, Ips pini (Say), often co-exist in lodgepole pine, Pinus contorta var. latifolia Engelmann. Intra- and interspecific semiochemical communication occurs in both species and their complete semiochemical repertoire and precise dynamics of pheromone production have not been elucidated. Porapak-Q extracts of captured volatiles from beetles of each species aerated at different attack phases (freshly emerged, pioneer sex alone in the log and both sexes paired in new galleries), followed by gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectroscopic analyses identified 17 compounds (seven compounds common to both species, six present in D. ponderosae and four present in I. pini) that excited the antennae of either or both species. Seven compounds for D. ponderosae and nine for I. pini had not been assessed for behavioural activity. In field trapping experiments, 2-phenylethanol produced by both species inhibited the response of D. ponderosae to its aggregation pheromones. exo- and endo-Brevicomin produced by D. ponderosae significantly decreased the response of I. pini to its aggregation pheromone ipsdienol. Nonanal, a ubiquitous compound found in the volatiles of lodgepole pine, various nonhosts and in both beetle species deterred the response of I. pini to ipsdienol. The occurrence of cis-verbenol, trans-verbenol and verbenone in emergent I. pini, and verbenone and 2-phenylethanol in emergent D. ponderosae suggests that these compounds may inhibit aggregation and induce dispersal following emergence. Termination of aggregation in D. ponderosae appears to depend on the production of frontalin in combination with changes in the relative ratios of verbenone, exo-brevicomin, trans-verbenol and 2-phenylethanol. In I. pini, the cessation of ipsdienol production by males is probably the main factor in terminating aggregation. Received 16 November 1999; accepted 7 August 2000     

Rutabaga roots, a rich source of oviposition stimulants for the cabbage root fly

Summary. Two recently identified compounds ("CIFs"), present on the leaf surface of Brassica oleracea (cabbage), are the strongest oviposition stimulants known for the cabbage root fly, Delia radicum. Cabbage leaves contain these compounds in extremely low concentrations, and the amount of CIFs obtained from purifying leaf extracts was so small that it limited further research. We were able to purify far more of these two compounds from the roots of Brassica napus var. napobrassica (rutabaga). Apart from being a richer source of CIFs, rutabaga roots are considerably easier to collect and process than leaves. In addition, we isolated and identified a new CIF compound from the roots that is also very active in stimulating oviposition in the cabbage root fly. Received 26 May 2000; accepted 26 July 2000     

Lignoid chemical defenses in the freshwater macrophyte Saururus cernuus

Summary. Chemical defense against herbivores has rarely been investigated for freshwater plants, possibly due to the common misconception that herbivory on aquatic macrophytes is low and would not select for chemical defenses. In previous work, the freshwater angiosperm Saururus cernuus was shown to be a low preference food for omnivorous crayfish despite its high nutrient value and relatively soft texture. We used feeding by the crayfish Procambarus clarkii to guide fractionation of the deterrent lipid-soluble extract of this plant, leading to the identification of seven deterrent lignoid metabolites, (–)-licarin A, (+)-saucernetin, (–)-dihydroguaiaretic acid, (–)-sauriols A and B, (–)-saucerneol, and (–)-saucerneol methyl ether. Lignans have been implicated in terrestrial plant chemical defenses as insect growth inhibitors, insect toxins, nematocides, antibacterial, and antifungal agents. However, these activities have rarely been demonstrated using ecologically relevant methodologies in terrestrial systems, and never before in freshwater systems. The widespread nature of lignans amongst very distantly related plants, along with their rich diversity of molecular structure, suggests that they could play a large role in mediating plant-herbivore interactions. In addition to the lignoid compounds we identified, there were other compounds present in low concentration or unstable compounds that were deterrent, that did not appear to be lignans, but that we were unable to identify. This plant thus appears to be defended by a complex mixture of natural products. Received 6 June 2000; revised 23 August 2000; accepted 2 September 2000     

Spread of metals through an invertebrate food chain as influenced by a plant that hyperaccumulates nickel

Summary. Hyperaccumulation of metals in the shoot system of plants is uncommon, yet taxonomically and geographically widespread. It may have a variety of functions, including defense against herbivores. This study investigated the effects of hyperaccumulation on metal concentrations across trophic levels. We collected plant material, soil, and invertebrates from Portuguese serpentine outcrops whose vegetation is dominated by the nickel hyperaccumulator Alyssum pintodasilvae. Samples were analyzed for nickel, chromium, and cobalt. Grasshoppers, spiders, and other invertebrates collected from sites where A. pintodasilvae was common had significantly elevated concentrations of nickel, compared to nearby sites where this hyperaccumulator was not found. Chromium and cobalt, occurring in high concentrations in the serpentine soil but not accumulated by A. pintodasilvae, were not elevated in the invertebrates. Therefore, it appears likely that a flux of nickel to herbivore and carnivore trophic levels is specifically facilitated by the presence of plants that hyperaccumulate this metal. The results may be relevant to the development of phytoremediation and phytomining technologies, which use plants to extract metals from the soil. Reveived 22 August 2002; accepted 2 April 2003. R1D=" Correspondence to: A. J. Pollard, e-mail:joe.pollard@furman.edu     

Induction of host specificity in larvae of Manduca sexta: chemical dependence controlling host recognition and developmental rate

Summary. The tobacco hornworm, Manduca sexta L. (Lepidoptera: Sphingidae) is a specialist on Solanaceae. This host specificity is induced as the larva feeds on solanaceous foliage, so that solanaceous-reared larvae will refuse to feed on other plants. Experiments were designed to determine the role of dietary constituents on the induction of host specificity and the effects of these on development of M. sexta. Choice assays using leaf discs of cowpea, Vigna sinensis (Fabaceae), were used to monitor the isolation of relevant chemical cues from foliage of potato. An aqueous extract of potato foliage strongly stimulated feeding. This extract was partitioned with n-butanol under alkaline conditions to obtain a highly active butanol extract. Reversed phase flash chromatography with a water-methanol gradient gave an active fraction that was used as a supplement for wheat germ-based artificial diet. Larvae reared on this s-diet became dependent on potato allelochemicals for initiation and continuation of feeding activity. These larvae also developed faster than larvae reared on the control p-diet, but no effect on adult mass was detected. Further flash chromatography of the active fraction under alkaline conditions provided a highly active sub-fraction, and semi-preparative HPLC using gradients of water and acetonitrile resulted in the isolation of a single compound. Bioassays indicated that this compound alone can account for host recognition by solanaceous-reared larvae. The results suggest that the mechanism of induced host specificity in M. sexta involves development of dependence on this compound. Received 21 December 1999; accepted 14 March 2000     

Differences in the electroantennal responses of apple- and hawthorn-infesting races of Rhagoletis pomonella to host fruit volatile compounds

Summary. Domestic apple (Malus pumila)- and hawthorn (Crataegus sp.)-infesting races of Rhagoletis pomonella, Walsh (Diptera: Tephritidae) provide an excellent model to examine the role that host plant specificity plays during sympatric speciation (i.e., divergence in the absence of geographic isolation). Previous work has shown that these races differ in their propensities to accept apple and hawthorn fruits in behavioral choice assays, and that this discrimination translates into "host fidelity" in the field (i.e., apple flies tend to mate on and oviposit into apples and hawthorn flies on hawthorns). ?We present the results of a study examining possible physiological factors contributing to host choice differences in R. pomonella. We tested whether apple and hawthorn flies differ in their electroantennogram (EAG) responses to biologically relevant volatile compounds emitted from apples and hawthorns. Significant differences were found in the relative EAG responses of apple and hawthorn flies to host fruit compounds at five of six paired study sites across the eastern United States. The geographic pattern of EAG variation was complex, however, with local populations of apple and hawthorn flies tending to be more similar to one another than to flies of the same race at distant sites. This pattern was largely due to EAG responses for several compounds showing longitudinal or latitudinal clines, the latitudinal clines being similar to those observed for allozyme loci in the host races. We also found evidence for sex-related differences, as males tended to have higher mean EAG responses to compounds than females. Host-associated differences were therefore nested within geographic and sex-related differentiation in R. pomonella.?Further behavioral studies are needed to distinguish whether the EAG differences are responsible for, as opposed to being a consequence of, host-plant fidelity and adaptation. Crosses are also required to establish a genetic basis for the EAG responses, although we did find significant correlations between EAG scores for several compounds and the allozymes NADH-Diaphorase-2 and Hydroxyacid dehydrogenase at one of the study sites. Questions therefore remain concerning the evolutionary significance of the EAG response differences between apple and hawthorn fly races. Nevertheless, these differences raise the possibility that antennal responses to fruit-related volatile compounds contribute to host plant discrimination in R. pomonella. Regardless, the EAG responses represent another set of traits, in addition to diapause/eclosion time phenotypes and allozyme frequencies, differing between apple and hawthorn host races of R. pomonella. Received 17 March 1998; accepted 21 September 1989.     

Oviposition stimulant for the zebra swallowtail butterfly, Eurytides marcellus, from the foliage of pawpaw, Asimina triloba

Summary. We have isolated a caffeoylcyclohexane-1-carboxylic acid derivative, 3-caffeoyl-muco-quinic acid (3-CmQA), as a contact oviposition stimulant for the zebra swallowtail butterfly, Eruytides marcellus (Papilionidae), from the foliage of its primary host plant, Asimina triloba (Annonaceae). This compound alone was as active in stimulating oviposition by females as were the parent ethanolic plant extract and the host plant itself. Other tested isomers of 3-CmQA, including 5-caffeoylquinic acid (5-CQA or trans-chlorogenic acid), were inactive. We found, however, that experienced female butterflies responded strongly to host volatiles, which enhanced landing rates and hence oviposition.? This is the first report of an oviposition stimulant for a swallowtail butterfly of the tribe Graphiini. We found 3-CmQA to be the major caffeoylcyclohexane-1-carboxylic acid isomer in plants of the genus Asimina. These plants lack appreciable amounts of 5-CQA, which has been shown previously to be one of the oviposition stimulants for certain Rutaceae- or Apiaceae-feeding swallowtails of the related tribe Papilionini.? Our findings, along with earlier results from the tribes Troidini and Papilionini, suggest that responses by swallowtails to hydroxycinnamic acid derivatives as oviposition cues date back at least to the ancestor of the subfamily Papilioninae. Received 24 March 1998; accepted 27 May 1998.     

Chemical trail communication in the amblyoponine species Mystrium rogeri Forel (Hymenoptera, Formicidae, Ponerinae)

Summary. Workers of the amblyoponine species Mystrium rogeri employ trail communication during recruitment to food sources and new nest sites. The trail pheromone originates from a hitherto unknown sternal gland located in the 7th abdominal sternite. The recruiting ant deposits the gland secretions by a special gaster-dragging behavior. The recruitment behavior can be complemented by a rapid vertical body shaking performed by some recruiting ants inside the nest. M. rogeri workers possess a large pygidial gland, the secretion of which elicits a repellent response in other ant species. Received 25 May 1998; accepted 15 June 1998.