Chemical defense of a primitive Australian bombardier beetle (Carabidae):Mystropomus regularis

Summary The Australian bombardier beetle,Mystropomus regularis, sprays a mixture of quinones (1,4-benzoquinone, 2-methyl-1,4-benzoquinone, 2-ethyl-1,4-benzoquinone) and hydrocarbons (principallyn-pentadecane). The defensive fluid ist generated explosively in two-chambered glands, and is ejected audibly and hot (maximal recorded temperature = 59°C).Mystropomus is a member of the paussoid lineage of bombardiers. In common with other members of the group, it has a pair of elytral flanges (flanges of Coanda), associated with the gland openings, that serve as launching guides for anteriorly-aimed ejections of spray. It is argued thatMystropomus may be the least derived of flanged paussoids, and the closest living relative of the most primitive of extant bombardiers (Metriini).Paper no. 101 of the series Defense Mechanisms of Arthropods; no. 100 is Attygalleet al. J Chem Ecol 17: 805 (1991)     

Pheromones and colonization: reassessment of the milkweed bug migration model (Heteroptera: Lygaeidae: Lygaeinae)

Summary. Research on insect migration has justifiably emphasized females – the so-called “oogenesis-flight syndrome”– since it is the females that place the eggs into new habitats. The large and small milkweed bugs, Oncopeltus fasciatus and Lygaeus kalmii, respectively, have featured prominently in studies of insect migration and sequestration of host plant toxins for chemical defense. Here we report that males of these species, and males of another well-studied lygaeine (Neacoryphus bicrucis), produce pheromones in glands usually considered to serve only a defensive role in Heteroptera (the metathoracic scent glands), and that these pheromones are exploited by a tachinid parasitoid as a host-finding kairomone. The pheromones are mixtures of C₆ and C₈ saturated and unsaturated esters reminiscent of lepidopteran pheromones, and the key compound of the O. fasciatus pheromone has now been correctly identified as (E)-2,7-octadienyl acetate. It is proposed that the concept of the oogenesis-flight syndrome for these kinds of insects should accommodate the role of males in the migration process. The hypothesis is presented that male-produced pheromones play a significant role in guiding colonization of new habitats in many heteropteran species. In addition, data are presented suggesting that there is a trade-off between the amount of pheromone produced by colonizing males and the host breadth of the species. Received 21 December 1998; accepted 15 February 1999.     

Hydroquinones from defensive secretion of a giant Pacific millipede, <Emphasis Type="Italic">Acladocricus setigerus</Emphasis> (Diplopoda: Spirobolida)

Summary. The presence of considerable quantities of hydroquinones including hydroquinone, 2-methylhydroquinone, 2,3- dimethoxyhydroquinone, 2-methoxy-3-methylhydroquinone, and 2,3-dimethoxy-5-methylhydroquinone renders the defensive secretion of Acladocricus setigerus (Silvestri, 1897) significantly different from that of other quinone-producing millipedes. In addition, two hitherto undescribed compounds, namely, 2,3-dimethoxy-5-methylhydroquinone and 2-methyl-3,4-methylenedioxyphenol, were characterized from the defensive secretion. However, it is uncertain if the latter compound is formed after the release of the secretion. The two major compounds, 2-methyl-1,4-benzoquinone and 2-methoxy-3-methyl-1,4-benzoquinone, constitute about 75% of the defensive fluid. Furthermore, hydrocarbons, which are typically present in the secretions of most other arthropods that use benzoquinones as repellents, are notably absent in the secretion of A. setigerus.     

Spray mechanism of crepidogastrine bombardier beetles (Carabidae; Crepidogastrini)

Summary.   The defensive glandular apparatus of primitive bombardier beetles of the tribe Crepidogastrini (Carabidae) is described for the first time. As exemplified by two African species (Crepidogaster ambreana and C. atrata), the apparatus conforms to the basic bombardier plan, in that the glands are bicompartmented and the secretion is quinonoid (it contains 1,4-benzoquinones and hydrocarbons), hot, and discharged audibly. In a number of morphological respects the crepidogastrine apparatus resembles that of the classical bombardiers of the tribe Brachinini (rather than that of bombardiers of the paussoid lineage), reinforcing the view, already held on taxonomic grounds, that the Crepidogastrini and Brachinini are closely related. That the Crepidogastrini may be primitive relative to Brachinini is underscored by the finding that, unlike brachinines, crepidogastrines do not pulse their secretory emissions. Moreover, they discharge their secretion as a mist, rather than forcibly in the form of jets. Received 22 May 2001; accepted 29 May 2001.     

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     

Pyrrolizidine alkaloids and pentacyclic triterpene saponins in the defensive secretions of Platyphora leaf beetles

Summary. Field collected exocrine defensive secretions of nine neotropical Platyphora species were analyzed for the presence of plant acquired pyrrolizidine alkaloids (PAs) and pentacyclic triterpene saponins. All species secrete saponins. In addition, five species feeding on Tournefortia (Boraginaceae), Koanophyllon (Asteraceae, tribe Eupatorieae) and Prestonia (Apocynaceae) were shown to sequester PAs of the lycopsamine type, which are characteristic for species of the three plant families. The PA sequestering species commonly store intermedine, lycopsamine and their O^3′-acetyl or propionyl esters as well as O⁷- and O⁹-hydroxyisovaleryl esters of retronecine. The latter as well as the O^3′-acyl esters were not found in the beetles’ host plants, suggesting the ability of the beetles to esterify plant derived retronecine and intermedine or its stereoisomers. Despite the conformity of the beetles’ PA patterns, considerable inconsistencies exist regarding the PA patterns of the respective host plants. One host plant was devoid of PAs, while another contained only simple necines. Since the previous history of the field collected beetles was unknown this discrepancy remains obscure. In contrast to the Palearctic chrysomeline leaf beetles, e.g. some Oreina species which ingest and store PAs as their non-toxic N-oxides, Platyphora leaf beetles absorb and store PAs as the toxic free base (tertiary PA), but apparently avoid to accumulate PAs in the haemolymph. This suggests that Chrysolina and Platyphora leaf beetles developed different lines of adaptations in their parallel evolution of PA mediated chemical defense. Received 30 November 2000; accepted 5 February 2001     

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.     

Activity of insect juvenile hormone III: seed germination and seedling growth studies

Summary. Juvenile hormones are sesquiterpenoids that regulate developmental processses such as metamorphosis and reproduction in insects. Insect juvenile hormone III (JH III), methyl-10R,11-epoxy-3,7,11-trimethyl-2E,6E-dodecadienoate, has also been identified in two sedge species, Cyperus iria and C. aromaticus (Toong et al. 1988). Potential allelopathic activity of this compound and the structurally related sesquiterpenoid farnesol was investigated using seed germination and seedling growth assays with radish, lettuce and rice. Treatment of seeds with JH III delayed lettuce seed germination and potently inhibited rice shoot growth. Both farnesol and JH III inhibited the growth of C. iria seedlings. The antimicrobial activity of JH III was also tested on a taxonomic and ecologically diverse range of fungi. Using the classic cytotoxic disk assay, JH III did not effect growth of the fungal species tested. We believe that JH III may contribute to the aggressive nature of this invasive weed species. Received 11 October 1999; accepted 26 February 2000     

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     

Sneak in or repel your enemy: Dufour's gland repellent as a strategy for successful usurpation in the slave-maker Polyergus rufescens

Summary. The dulotic queen ant, Polyergus rufescens, must first penetrate a host colony and kill the resident queen in order to successfully founding a new colony. Successful usurpation by a newly mated queen predictably depends on a dual strategy. Although, it can sneak in by being “chemically insignificant” with respect to cuticular hydrocarbons, it may also need to deter prospective host-worker aggressors. Chemical analysis of Dufour's gland secretion of P. rufescens queens and workers by GS/MS revealed that queen secretion is typified by esters of butanoic acid and acetic acid, of which decyl butanoate comprises over 80%. Butanoates and acetates are also present in the workers' secretion, but these are of higher molecular weight, and octadecyl butanoate represents the major compound. Using synthetic mixtures of queen and worker Dufour's gland, we tested the hypothesis that these secretions modify the aggressive behavior of the host species Formica cunicularia>. The queen-like synthetic mixture significantly reduced aggression of the host workers towards alien conspecifics, but neither pentane nor the worker-like synthetic mixture showed this effect. Although Dufour's gland content of >Polyergus queens was suggested to function as an appeasement pheromone (Topoff et al. 1988; Mori et al. 2000), we hypothesized that it may in fact act as a repellent. In order to test this hypothesis we exposed starved F. cunicularia workers to a droplet of honey on a glass slide applied with one of the following compounds: decyl butanoate (queen major compound), octadecyl butanoate (worker main compound), limonene (a reported ant repellent), and pentane (solvent control). Of these, the workers were repelled only by the decyl butanoate and did not approach the honey. We conclude that during usurpation the queen actively repels aggressive workers by emitting Dufour's gland repellent, comprising the alternative tactic in the usurpation dual strategy. This represents another chemical weapon in the diverse arsenal used by parasites to overcome the host's resistance. Received 7 April 2000; accepted 17 May 2000     

Different phagostimulants in potato foliage for Manduca sexta and Leptinotarsa decemlineata

Summary. In the past decades, several studies have focused on the identification of feeding stimulants for specialists that feed on solanaceous plants, especially potato (Solanum tuberosum). In the 1950's, a phagostimulant was isolated from potato and tomato for Manduca sexta and characterised as a glycoside. It was suggested that the phagostimulant for M. sexta and Leptinotarsa decemlineata is identical. We tested whether these insects indeed share the same recognition factor. Previous bioassays for L. decemlineata larvae and adults were found to be unsatisfactory, so a new assay system was developed, using starch and wheat flour-based wafers as a neutral substrate. An aqueous extract of potato foliage that was highly active as a stimulant was fractionated by reverse phase medium pressure liquid chromatography (MPLC). Both insects were stimulated by a MPLC fraction that eluted with 25% methanol in water. Further separation of this and the following fraction (35% methanol, stimulatory for the beetle only) by semi-preparative high performance liquid chromatography (HPLC), using gradients of water and acetonitrile, resulted in the isolation of two distinct fractions that stimulated feeding by L. decemlineata. None of the HPLC fractions was active for M. sexta. However, fractions of the flash chromatography with less polarity (45–75% methanol) had a stimulatory effect only on M. sexta. Thus, the two insects do not use a common feeding stimulant, and for both at least two compounds of different polarity are active. Received 3 July 2000; accepted 24 October 2000     

Sequestration of phorbolesters by the aposematically coloured bug Pachycoris klugii (Heteroptera: Scutelleridae) feeding on Jatropha curcas (Euphorbiaceae)

Summary. The phorbol ester DHPB has been detected in 5th instars and adults of Pachycoris klugii which feed on Jatropha curcas, a producer of phorbol esters with mollusc-, insect-, and vertebrate toxicity. DHPB from Pachycoris activates protein kinase C (PKC) which appears to be the main molecular target for phorbol esters. Phorbol esters of J. curcas exhibit a wide range of acute toxic effects in vertebrates and insects. It is therefore likely that the sequestration of DHPB, which would explain the aposematic colouration of the bugs, confers chemical protection to P. klugii against vertebrate predators. Received 26 April 2000; accepted 31 May 2000     

Molecular studies on the ouabain binding site of the Na+, K+-ATPase in milkweed butterflies

Summary. The Na⁺, K⁺-ATPase of the Monarch butterfly (Danaus plexippus) is insensitive to the inhibition by cardiac glycosides due to an amino acid replacement: histidine instead of asparagine at position 122 of the α-subunit representing the ouabain binding site. By PCR amplification of the DNA sequence of this site, a PCR product of 270 bp was obtained from DNA extracted from Danainae species (Danaus plexippus, D. chrysippus, D. gillipus, D. philene, D. genutia, Tirumala hamata, Euploea spp., Parantica weiskei, P. melusine), Sphingidae (Daphnis nerii) and mimics of milkweed butterflies (Hypolimnas missipus, Limenitis archippus and L. arthemis, Nymphalidae). Analysis of the nucleotide sequences revealed that the single point mutation in the ouabain binding domain (AAC-Asn for CAC-His) was present only in Danaus plexippus, but not in the other species investigated. Since these milkweed butterflies also store cardenolides, other structural modifications of the Na⁺, K⁺-ATPase may have occurred or other strategies of cardenolide tolerance have been developed. Received 15 May 2000; accepted 29 June 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     

Chemical composition and pheromonal function of the defensive secretions in the subtribe Stizopina (Coleptera, Tenebrionidae, Opatrini)

The chemical composition of the defensive secretions of 52 species from 15 genera of the tenebrionid subtribe Stizopina was analyzed. The secretions of all species contained 1,4-benzoquinones, 1-alkenes, and monoterpene hydrocarbons, only one species was lacking the latter. Methyl- and ethyl-1,4-benzoquinone were ubiquitous, mostly accompanied by smaller amounts of 1,4-benzoquinone as well as isopropyl- and propyl-1,4-benzoquinone. 1-Alkenes were dominated by 1-undecene with varying admixtures of other 1-alkenes. The monoterpene hydrocarbons always consisted of a mixture of α-pinene, camphene, β-pinene and limonene, but also p-cymene, α-terpinene or α-phellandrene were found in some species. Furthermore, some species synthesized additional compounds such as phenols, ketones, 2,5-dihydroxy-6-methylbenzoate, 2-hydroxy-4-methoxyacetophenone and naphthoquinones. Bioassays showed that the defensive secretion co-functioned as an aggregation pheromone in the subtribe Stizopina. All nine tested species from six genera were attracted to defensive secretion of Stizopina species, but they did not distinguish between defensive secretions of different Stizopina species. This lack of discrimination might be the initial step for the formation of interspecific aggregations and the evolution of cleptoparasitism within the Stizopina.     

Social feeding in ladybird beetles: adaptive significance and mechanism

Summary. Young larvae of Adalia bipunctata search an area more intensively when exposed to the odour from other larvae feeding on aphids than when exposed only to the odour of aphids. In an olfactometer young larvae were significantly attracted either to the odour of crushed aphids or larvae feeding on aphids, but not to that of aphids, larvae, larvae plus aphids or larvae feeding on an artificial diet. That is, the change in searching behaviour appears in response to a volatile released by aphids when attacked. The odour released by crushed aphids is made up entirely of aphid alarm pheromone, β-farnesene. It is likely that the adaptive significance of this response is that it increases the ability of larvae to locate larvae that have already caught prey. By sharing the aphid kill of another larva it is likely that a first instar ladybird larva greatly increases its probability of surviving to the next instar. It is suggested that this social feeding is facilitated by egg clustering, which also may additionally account for why aphidophagous ladybirds lay their eggs in clusters. Received 28 February 2000; accepted 24 March 2000     

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.     

A sex pheromone component novel to Ostrinia identified from Ostrinia latipennis (Lepidoptera: Crambidae)

Summary. Extracts from the sex pheromone gland of Ostrinia latipennis (Lepidoptera: Crambidae) were analyzed by gas chromatography-electroantennographic detection (GC-EAD) and GC-mass spectrometry. Only an EAD-active compound was detected in the extract, and it was identified as (E)-11-tetradecenol (E11-14:OH). In a wind-tunnel bioassay, E11-14:OH elicited a series of mate finding behaviors from males, although it was far less active than virgin females and crude extract of the pheromone gland. The attractiveness of E11-14:OH to O. latipennis males was confirmed by field trapping experiments. Based on these findings, we concluded that E11-14:OH, which is novel to the genus Ostrinia, is a major component of the sex pheromone in O. latipennis. The significance of the use of alcohol in place of the usual acetates in Ostrinia is discussed in relation to the pheromone biosynthesis system. Received 9 December 1999; accepted 14 March 2000     

Metabolic transformations of acquired lucibufagins by firefly “femmes fatales”

Summary. Female Photuris fireflies sequester defensive steroidal pyrones (lucibufagins) from male fireflies of the genus Photinus. Lucibufagin analyses of Photuris females and Photinus ignitus males show that the lucibufagin mixtures of predator and prey differ in their composition. Analyses of whole body extracts showed that P. ignitus males contain a mixture of eight non-glycosylated lucibufagins, composed mostly of compounds with two oxygenated positions in the steroidal A-ring (C-3, C-5). After feeding on P. ignitus males, Photuris females contain six major lucibufagins. Three of these compounds are not present in the prey, including the novel lucibufagin glycoside 5β,11α-dihydroxy-12-oxo-3β-O-β-D-xylopyranosylbufalin, and two other lucibufagins with a trioxygenated A-ring (C-2, C-3, C-5). These results indicate that Photuris females transform the sequestered lucibufagins both by glycosylation and oxidation, which could affect the systemic transportability of these compounds due to an increase in their polarity. Received 18 February 1999; accepted 19 April 1999.     

Defensive volatile secretions of two diplopod species attract the carrion ball roller scarab <Emphasis Type="Italic">Canthon morsei</Emphasis> (Coleoptera: Scarabaeidae)

Summary. We report field and laboratory evidence indicating that the defensive compounds secreted by the millipedes Amplinus bitumidus (Diplopoda: Polydesmida) and Anadenobolus putealis (Diplopoda: Spirobolida) attract the carrion scarab Canthon morsei (Scarabaeidae: Scarabaeinae). In a Mexican tropical rain forest, C. morsei was mostly caught in pitfall traps baited with the carcasses of both millipede species. Only a few beetles were attracted to fish carrion. Experimental evidence indicates that C. morsei is attracted to freshly dead millipedes. Benzaldehyde and HCN were identified from Amplinus bitumidus; but 2-methyl-1,4-benzoquinone (toluquinone) and 2-methoxy- 3-methyl-1,4-benzoquinone from Anadenobolus putealis. Benzaldehyde attracted significantly more C. morsei individuals than fish meat, or millipedes of both species did after rinsing in hexane. This is the first report of Polydesmida attracting dung beetles.