Defensive role ofAllium sulfur compounds for leek mothAcrolepiopsis assectella Z. (Lepidoptera) against generalist predators

It has been shown previously that sulfur volatiles produced byAllium plants affect the behavior of their specialist phytophages and of their specialist entomophages. The action of these compounds in protecting the leek mothAcrolepiopsis assectella against generalist entomophages was studied in comparison to the proposed original defensive role of these compounds against generalist herbivorous insects. Two ants species,Formica selysi andF. fusca, were used as generalist predators. Six behavioral criteria of the predatory behavior of the ants were studied in presence of the last-instar caterpillars (C₅). C₅ reared on artificial diets with or without leek components were tested, as well as C₅ soaked in frass of leek-reared caterpillars or disulfide solutions. In addition, the response of the ants to pure chemicals found in leek was studied using honey solutions with or without sulfur compounds. The sulfur allelochemicals ofAllium plants have a negative action on predatory ants. Interestingly, the nonvolatile precursors of sulfur volatiles ofAllium plants seem to have a protective role for their phytophagous insects against generalist entomophages.     

Chemical Ecology of the Defense of Two Nymphalid Butterfly Larvae Against Ants

We analyzed the behavioral responses of the ants Camponotus rufipes and Solenopsis geminata towards all instars of Dione junio and Abananote hylonome. We also analyzed ant behavior towards hexane extracts of larvae and extracts of the spines and neck glands of the fifth instars of both species and identified the chemical compounds present. Larvae of both species were repellent to ants from the first instar onward. Later instars survived ant attacks better than earlier instars. The spines and neck glands of the larvae influenced the behavior of C. rufipes. The chemical compounds contained in the hexane extracts of whole first and fifth instars and in the spines and neck glands of fifth instars were principally carboxylic acids and terpenes. Further bioassays confirmed the repellent effect of some of these acids toward ants.     

Effect of steam distillate extracts of selected resistant cultivated and wild rices on behavior of leaffolderCnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae)

Rice plant volatiles extracted as steam distillates significantly affected the behavior of the rice leaffolder,Cnaphalocrocis medinalis (Guenée). Leaffolder moths laid significantly fewer eggs on TN1 rice plants treated with extracts of resistant wild species of rice,Oryza officinalis andOryza punctata, than on TN1 plants treated with extracts of resistant cultivated rices. Extracts ofO. officinalis andO. punctata adversely affected egg hatchability.O. officinalis andO. punctata extracts were more toxic to first-instar larvae than extracts of other resistant varieties. Leaf area consumed by the larvae was reduced on TN1 plants treated withO. officinalis andO. punctata extracts compared with that on plants treated with extracts of resistant cultivated rice varieties.     

Toxicity of nitro compounds fromLotus pedunculatus to grass grub (Costelytra zealandica) (Coleoptera: Scarabaeidae)

Crude extracts of root of the forage legumesLotus pedunculatus andCoronilla varia (crownvetch) were toxic when administered orally to 3rd instarCostelytra zealandica larvae. A group of 3-nitropropanoyl-d-glucopyranoses was isolated from active fractions of the crude extracts. These toxins, some of which were already known fromC. varia, have not previously been reported fromL. pedunculatus. The compounds were present in root tissue of this species at a concentration of about 1% dry weight. They include the triester karakin and the diesters coronarian and cibarian, all of which were toxic to larvae ofC. zealandica at levels which could account fully for the activity of the crude extracts.     

Defensive adaptations of eggs and adults ofGastrophysa cyanea (Coleoptera: Chrysomelidae)

Egg clusters and adults ofGastrophysa cyanea are conspicuous and, like their larvae, are chemically protected. The eggs owe their bright yellow color primarily to -carotene and, in addition, contain substantial quantities of oleic acid. At natural concentrations oleic acid effectively deters many species of ants from feeding. The use of fatty acids as deterrents against ants is discussed as a possible widespread phenomenon among insects. During defensive confrontations, adults ofG. cyanea exhibit avoidance behavior and may also feign death. In addition, the adults may autohemmorhage or secrete a fluid from elytral or pronotal pores in response to traumatic stimuli. The secretions are effective against ants and contain a mixture of hydrocarbons as well as terpenoid components. The pattern of ontogenetic modification in the defensive chemical repertoire ofG. cyanea is discussed.     

Sequestration ofVeratrum alkaloids by specialistRhadinoceraea nodicornis konow (Hymenoptera,Tenthredinidae) and its ecoethological implications

The larvae of the specialist sawflyRhadinoceraea nodicornis Konow (Hymenoptera, Tenthredinidae) store in their hemolymph ceveratrum alkaloids originating from the host plantVeratrum album L. (Liliales, Melanthiaceae). The major alkaloid found in the hemolymph is 3-acetyl-zygadenine. Qualitative and quantitative data showed that the plant alkaloid 3-angeloylzygadenine is most probably metabolized in the larval gut to zygadenine and then acetylated. A still unidentified alkaloid with a molecular weight of 591 Da was detected in plant leaves as well as in the gut, hemolymph, and excrement of larvae. Protoveratrine A and B, on the other hand, seem to be degraded by the larvae. These findings indicate that the pathway of ceveratrum alkaloids inR. nodicornis larvae is fourfold: direct sequestration, metabolism followed by sequestration, excretion of intact alkaloids, and degradation. In contrast, no ceveratrum alkaloids were detected in the hemolymph and excrement of larvae of the generalist sawflyAglaostigma sp. fed withV. album leaves. Bioassays with the antMyrmica rubra L. proved that the hemolymph ofR. nodicornis larvae is highly deterrent and toxic. In bioassays evaluating defensive efficiency against predators (ants, spiders, and bushcrickets), no larvae were eaten. Ceveratrum alkaloids were also detected in the hibernating prepupae ofR. nodicornis. In feeding bioassays, the shrewCrocidura russula Hermann rarely fed upon prepupae, suggesting that this stage is also protected from predation to some degree. In field surveys, the only parasitoids recorded were two ichneumonid species that are believed to be specialized onR. nodicornis. Bioassays and field observations enable us to suppose thatR. nodicornis and its enemies produce a food web of ion connectance.     

Crude extracts of asteraceous weeds

Petrol and ethanolic extracts of six asteraceous weeds were added to artificial diets to screen for growth inhibition and mortality of the variegated cutworm,Peridroma saucia (Hbn). Petrol and ethanolic extracts ofArtemisia tridentata andChamomilla suaveolens and ethanolic extracts ofChrysothamnus nauseosus andCentaurea diffusa severely inhibited larval growth at five times the natural concentrations. The twoC. suaveolens extracts and the ethanol extract ofA. tridentata were active at the natural concentration (100%) and were further examined at 20, 40, 60, and 80% of this level. Inhibition of larval growth was directly related to concentration for each of the three extracts tested. EC₅₀s (effective concentration to inhibit growth by 50% relative to controls) for the three extracts were 36–42% of the naturally occurring level in the plants. Nutritional indices were calculated for secondinstarP. saucia feeding on the activeA. tridentata EtOH extract and the petrol extract fromC. suaveolens. Addition of the activeA. tridentata EtOH or theC. suaveolens petrol extract to the diet resulted in significant reduction in the relative growth rate of larvae, although theA. tridentata extract was much more inhibitory. Dietary utilization was significantly lower for larvae fed theA. tridentata EtOH extract.     

Composition of larval secretion ofChrysomela lapponica (Coleoptera,Chrysomelidae) and its dependence on host plant

The defensive secretion ofChrysomela lapponica larvae, which is produced by nine pairs of exocrine dorsal glands, has been chemically analyzed. TheC. lapponica larvae were kept in the laboratory on leaves of either birch (Betula pendula), alder (Alnus glutinosa), or willow (Salix fragilis). Larvae developed normally on birch and willow, whereas those on alder died within a few days. GC-MS analyses of the secretion of larvae on birch and willow revealed that the composition of this secretion differs distinctly from the known ones of several otherChrysomela species feeding exclusively on Salicaceae. In the exocrine secretion of larvae on birch, 69 compounds were identified, which included the main components isobutyric acid, 2-methylbutyric acid, and esters of the two. Several of the esters have not been reported previously from nature. The alcoholic components of the esters may be hydrolysis products ofBetula glycosides. Most components of the secretion of larvae feeding on birch were also found in the secretion of larvae feeding on willow. In addition, major amounts of benzoic acid and salicylalcohol were present in the secretion of the larvae feeding on willow.C. lapponica obviously acquires salicylalcohol by hydrolysis of salicin from willow leaves. However, in contrast to otherChrysomela species,C. lapponica larvae oxidize only traces of salicylalcohol to salicylaldehyde. The repellent activity of single authentic compounds of the secretion of larvae feeding on birch and willow, respectively, was tested in laboratory bioassays with ants (Myrmica sabuleti). Biosynthetic pathways to some identified compounds are suggested and discussed under evolutionary and functional aspects.     

Role of volatile inforchemicals emitted by feces of larvae in host-searching behavior of parasitoidCotesia rubecula (Hymenoptera: Braconidae): A behavioral and chemical study

The role of volatile infochemicals emitted by feces of larvae in the host-searching behavior of the parasitoidCotesia rubecula was evaluated during single- and dual-choice tests inside a wind tunnel. The following treatments were tested: feces produced by second and fourth instars ofPieris rapae (preferred host), second instars ofP. brassicae (inferior host), second instars ofP. napi (nonhost), and wet feces of second instars ofP. rapae. During a single-choice situation females ofC. rubecula oriented to all types of feces tested. When a preference was to be made,C. rubecula preferred feces of second instars ofP. rapae over that of fourth, feces ofP. rapae over that ofP. brassicae, feces ofP. napi over that ofP. brassicae, and wet over normal host feces. No preference was exhibited between feces of second instars ofP. napi and that of second instars ofP. rapae. The relative importance of infochemicals from host feces versus plant damage caused by host larvae to the searching behavior ofC. rubecula was also evaluated. Plant damage was more important to the searching females than host feces when feces were present in specific concentrations in relation to damage. The volatiles released by normal and wet feces of second instars ofP. rapae, wet feces of fourth instars ofP. rapae, and normal and wet feces ofP. brassicae were collected and identified. Overall, 85 chemical compounds were recorded belonging to the following chemical groups: alcohols, ketones, aldehydes, esters, isothiocyanates, sulfides, nitriles, furanoids, terpenoids and pyridines. The blend of chemicals emitted by feces of different instars ofP. rapae and different species ofPieris exhibited an instar and species specificity in both quantity and quality. Wetting of normal feces increased the amount of volatile chemicals released, and it was also responsible for the appearance of new compounds. The role of feces of larvae in the host-seeking behavior ofC. rubecula is discussed.     

Sticky secretion from two pairs of defensive glands of rove beetleDeleaster dichrous (Grav.) (Coleoptera: Staphylinidae)

The phylogenetically primitive rove beetleDeleaster dichrous (Grav.) (Oxytelinae) has been shown for the first time to possess two pairs of neighboring abdominal glands which are depleted simultaneously on molestation. The morphology of these glands is described. The defensive constituents of theDeleaster glands were elucidated directly from the mixtures by gas chromatographic-mass spectroscopic methods and microchemical reactions. The paired whitish glands secrete iridodial, which polymerizes on exposure to air to form an adhesive that probably deters small predatory arthropods. The red gland system ofD. dichrous contains the toxicp-toluquinone and a variety of isopropyl andsec-butyl esters. Artificial quinoid ester mixtures simulating the secretion ofD. dichrous showed only weak effects on mortality ofLucilia larvae in comparison with more effective secretions of phyiogenetically derived Oxytelinae. The secretion of the primitive genusDeleaster is characterized chemotaxonomically by , -unsaturated C₁₂ acids and esters, which are postulated as precursors for the characteristic defensive compounds of the derived species, thus indicating a clear evolutionary trend at the micromolecular level.     

Similarity of Cuticular Lipids Between a Caterpillar and Its Host Plant: A Way to Make Prey Undetectable for Predatory Ants?

Ithomiine butterflies (Nymphalidae) have long-lived, aposematic, chemically protected adults. However, little is known about the defense mechanisms in larvae and other juvenile stages. We showed that larvae Mechanitis polymnia are defended from ants by a chemical similarity between their cuticular lipids and those of the host plant, Solanum tabacifolium (Solanaceae). This is a novel defense mechanism in phytophagous insects. A field survey during one season showed that larval survivorship was up to 80%, which is high when compared with other juvenile stages. In a laboratory bioassay, live larvae on their host plant were not attacked by the predatory ant Camponotus crassus (Formicidae). Two experiments showed that the similarity between the cuticular lipids of M. polymnia and S. tabacifolium protected the larvae from C. crassus: (a) when the caterpillar was switched from a host plant to a non-host plant, the predation rate increased, and (b) when a palatable larva (Spodoptera frugiperda, Noctuidae) was coated with the cuticular lipids of M. polymnia and placed on S. tabacifolium leaves, it no longer experienced a high predation rate. This defensive mechanism can be defined as chemical camouflage, and may have a double adaptive advantage, namely, protection against predation and a reduction in the cost of sequestering toxic compounds from the host plant.     

Semiochemicals from anal exudate of larvae of tsetse fliesGlossina morsitans morsitans Westwood andG. morsitans centralis Machado attract gravid females

Tsetse flies (Diptera: Glossinidae:Glossina) mature their offspring in utero, giving birth to mature larvae that burrow into soil and pupariate. During the hot dry seasons, puparia of some species of tsetse are aggregated in areas of deep shade in dense thickets. We have confirmed the presence of a semiochemical from the prepupariation excretions of larvae ofGlossina morsitans morsitans Westwood and report a similar semiochemical inGlossina morsitans centralis Machado. These semiochemicals are attractive to gravid females and result in the aggregation of puparia. Behavioral studies withG. m. centralis showed that a higher percentage of females larviposited over moist sand conditioned by the anal exudate of larvae. Electroantennogram analyses of extracts of sand conditioned byG. m. centralis andG. m. morsitans confirmed the presence of olfactory receptors on the antennae for the semiochemicals. Both subspecies responded to extracts of the semiochemicals of the other, withG. m. morsitans more responsive to lower concentrations of extract ofG. m. centralis than the converse.     

Fate of iridoid glycosides in different life stages of the Buckeye,Junonia coenia (Lepidoptera: Nymphalidae)

The buckeye butterfly,Junonia coenia (Lepidoptera: Nymphalidae), specializes on plants that contain iridoid glycosides. To determine the fate of these compounds in larvae, pupae, and adults of this species, we reared larvae on artificial diets with and without iridoid glycosides, and on leaves of a host plant,Plantago lanceolata (Plantaginaceae). Quantification by gas chromatography showed that newly molted third-, fourth-, and fifth-instar larvae reared on leaves ofP. lanceolata contained means of 5.13, 2.88, and 6.83% dry weight iridoid glycoside. In contrast, the mean iridoid glycoside concentration of actively feeding fifth-instar larvae was 0.28% dry weight, that of pupae was 0.19% dry weight iridoids, and adults contained no detectable iridoids. Feeding experiments suggested that this reduction in actively feeding larvae was due to the metabolism of iridoid glycosides.P. lanceolata leaves in these experiments contained a mean of 1.00% dry weight iridoid glycoside, with a 2:1 ratio of aucubin to catalpol. Calculation of iridoid consumption and utilization indices showed that larvae fed artificial diets consumed, digested, and sequestered aucubin and catalpol in similar ways. When these indices were calculated for larvae fed leaves ofP. lanceolata, catalpol was sequestered twice as efficiently as aucubin.     

Recognition of aphid parasitoids by honeydew-collecting ants: The role of cuticular lipids in a chemical mimicry system

The aphidiid waspLysiphlebus cardui parasitizes in ant-attendedAphis fabae cirsiiacanthoidis colonies without causing aggressive behavior in the antLasius niger. By contrast,Trioxys angelicae, another aphidiid parasitoid of aphids, is rapidly recognized and vigorously attacked by the ants.L. niger workers also responded differently to dead individuals ofL. cardui andT. angelicae. DeadL. cardui parasitoids were often ignored when encountered byL. niger, whereas deadT. angelicae individuals were immediately grasped by ants that discovered them. However, hexane-washed parasitoids caused a similar reaction pattern in the ants, in that both aphidiid species were tolerated in the aphid colony. Lure experiments demonstrated that chemical stimuli on the cuticle are major cues for the ants to distinguish between the parasitoids. The hexane extract ofL. cardui transferred to washed individuals ofT. angelicae resulted in ant responses characteristic towardsL. cardui, andL. niger workers displayed the typical removal pattern they normally showed towardsT. angelicae whenT. angelicae extract was applied toL. cardui individuals. Both parasitoid species treated with the hexane extract ofA. fabae cirsiiacanthoidis were similarily treated by the ants as were aphid control individuals. The suggestion that the aphidiid waspL. cardui uses chemical mimicry is discussed.     

Trail and arena marking by caterpillars ofArchips cerasivoranus (lepidoptera: Tortricidae)

The activity ofArchips cerasivoranus caterpillars is largely limited to their colonial silk web and trails. Silk pulled directly from the spinnerets of caterpillars and wound onto paper strips to form artificial trails elicited locomotion from the larvae. Trails made from extracts of silk and silk glands also elicited locomotion. These and other observations reported here indicate that the caterpillars are responsive to a water-soluble pheromone that is a component of the silk strand. Marker pheromones appear not to be secreted from other regions of the body, as has been reported for some other trail-following caterpillars.     

Sequestration of pyrrolizidine alkaloids in several arctiid moths (Lepidoptera: Arctiidae)

Sequestration of dietary pyrrolizidine alkaloids (PA) by larvae and adults of six European arctiid moth species (Spilosoma lubricipeda, Arctia caja, Phragmatobia fuliginosa, Callimorpha dominula, Diacrisia sannio, andTyria jacobaeae) was investigated for comparison with the well-studied Asian arctiidCreatonotos transiens. Larvae of all species metabolized free PA bases into the respectiveN-oxides. Only adults ofA. caja, P. fuliginosa, andS. lubricipeda, but not their larvae, converted dietary 7(S)-heliotrine to 7(R)-heliotrine, a direct precursor of a male pheromone in some arctiids, 7(R)-hydroxydanaidal. The larval integument figures as the main storage site for resorbed alkaloids; only minor amounts were found in other tissues. In addition, a significant amount of alkaloid is deposited in the cocoon ofArctia caja; only traces of alkaloids could be found in the meconium and the exuviae of this species. A substantial part of the alkaloids fed was degraded to unknown, nonalkaloidal products.     

Attacus atlas caterpillars (Lep., Saturniidae) spray an irritant secretion from defensive glands

The ability ofAttacus atlas caterpillars to spray a defensive secretion seems to be due to the fine structure of the integumental glands that produce it. The giant gland cells are fixed to stable cuticular rings surrounding the gland openings and tightly closed by cuticular lids. Probably by increasing hemolymph pressure, the lids are blasted off and the secretion spouts out. The fluid contains several aromatics, biogenic amines (e.g., acetylcholine, histamine), glycerol, and trehalose and exhibits tyrosinase activity. Deterrent effects of caterpillar secretion and hemolymph on predatory ants could be shown. Presumably the spraying process serves to apply the secretion to sensitive sites of vertebrate target organisms.     

Developmental inhibition ofSpodoptera litura (Fab.) larvae by a novel caffeoylquinic acid from the wild groundnut,Arachis paraguariensis (Chod et Hassl.)

A novel compound, 1-caffeoyl-4-deoxyquinic acid (1-CdQA) has been identified along with 3-caffeoylquinic acid (3-CQA) and 5-caffeoylquinic acid (5-CQA) (syn. chlorogenic acid) in the foliage ofArachis paraguariensis, a wild species of groundnut that is highly resistant to attack by the larvae ofSpodoptera litura. When neonate larvae were fed on diets treated with 3-CQA or 1-CdQA, their development was severely inhibited compared to larvae on untreated diets, and the effects were similar in nature and magnitude to those observed for larvae feeding on diets treated with 5-CQA, rutin, and quercetin. The effects of all the compounds were dose related, and their optimal concentration was approximately 3 mM, which corresponds to the total foliar concentration of both caffeoylquinic acids and quercetin diglycosides inA. paraguariensis. After 24 h, the development of third stadium larvae feeding on diets treated with 5-CQA, rutin and 1-CdQA was promoted compared to larvae on control diets, but after 96 hr larvae feeding on treated diets had gained significantly less weight than those on the control diets. 1-CdQA, 3-CQA, and quercetin dyglycosides (previously identified in the foliage ofA. paraguariensis) are considered to be valuable components in resistance of groundnuts toS. litura and may provide useful genetic markers in future breeding for resistance to this pest.     

Mechanisms of resistance in wild riceOryza brachyantha to rice leaffolderCnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae)

A wild rice,Oryza brachyantha, was rated as highly resistant to rice leaffolder,Cnaphalocrocis medinalis (Guenée), while its F₁ hybrid from a cross with a high-yielding, leaffolder-susceptible rice, IR31917-45-3-2, was rated resistant in a standard screening test. In comparison with IR31917-45-3-2,O. brachyantha and the F₁ hybrid were unsuitable for oviposition, survival, and growth ofC. medinalis. However, growth and survival of larvae in artificial diets containing lyophilized leaf powder of IR31917-45-3-2 andO. brachyantha were comparable. Studies on the feeding and settling preference of the larval stages suggested that the mechanism of resistance ofO. brachyantha is that of antixenosis type. Olfactometer tests with first-instar larvae and electroantennogram responses of adults indicated a lack or low levels of volatile attractants inO. brachyantha and the F₁ hybrid in comparison with IR31917-45-3-2. Bioassays of sequential solvent extracts of these plants indicated that larval preference for IR31917-45-3-2 and nonpreference forO. brachyantha and the F₁ hybrid were due partly to chemical factors present in hexane and methylene chloride extractables. Closer arrangement of silica cells in the epidermal layer ofO. brachyantha and a higher mandibular wear in larvae reared onO. brachyantha suggested that physical resistance due to silica may be an additional cause of resistance. It was hypothesized that the high levels of resistance observed inO. brachyantha may be due to an additive or synergistic action of the absence of attractants or feeding stimulants, the presence of deterrents, and the physical resistance offered by silica. The significance of these results to a successful wide hybridization program aimed at transferring resistance factors from wild rice to cultivated rice is discussed.     

Growth inhibition ofMusca domestica L. andCulex quinquefasciatus (say) by (−)-3-epicaryoptin isolated from leaves ofClerodendron inerme (Gaertn) (Verbenaceae)

The petroleum ether extract ofClerodendron inerme leaves afforded a compound that matched the clerodane compound (–)-3-epicaryoptin in physical and spectral characteristics. The test compound inhibited the development ofMusca domestica andCulex quinquefasciatus larvae. First- and third-instar fly larvae reared on 3-epicaryoptin-treated diet pupariated later and earlier than their respective controls and adult emergence from puparia was inhibited. Apart from larval mortality, exposure of fourth-instar mosquito larvae to 3-epicaryoptin resulted in death at larval-pupal molt and pupaladult eclosion, indicating inhibition of the molting process.