Effect of oviposition deterrents from elderberry on behavioral responses byHeliothis virescens to host-plant volatiles in flight tunnel

In flight-tunnel assays, mated femaleHeliothis virescens (F.) moths responded by positive anemotaxis to volatiles from extracts of two host plants (cotton and tobacco), but they did not fly to an extract from elderberry (Sambucus simpsonii Rehd.), a nonhost that contains an oviposition deterrent forH. virescens. When the elderberry extract was mixed with extract from either cotton or tobacco, the flight response by moths to volatiles emanating from the extract blends was reduced significantly at most doses when compared to the positive response to extracts from either host alone. The number of landings (including brief contacts) and landings that resulted in oviposition on the substrates treated with extract blends also were reduced significantly in most tests.This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation of its use by USDA.     

Attraction ofHeliothis virescens (F.) (Lepidoptera: Noctuidae) to volatiles from extracts of cotton flowers

Mated and virgin female and virgin male tobacco budworm moths,Heliothis virescens (F.), responded via directed upwind flight in wind-tunnel bioassays to volatiles from floral extracts of cotton,Gossypium hirsutum L. Significantly more male and female moths landed on cloth dispensers treated with a methylene chloride extract of debracted flowers than on control dispensers treated only with methylene chloride. Only mated females landed in significant numbers on the dispensers treated with extract of cotton squares, flowers (including bracts), or flower petals. After landing, most of the moths examined the dispensers by probing with their antennae, proboscis, and/or ovipositor.     

Flight activity ofHeliothis virescens (F.) females (Lepidoptera: Noctuidae) with reference to host-plant volatiles

Heliothis virescens (F.) females responded positively via upwind flight in laboratory assays to volatiles emitted from methyle chloride washes of fresh whole leaves of host plants including cotton, tobacco, and a weed species,Desmodium tortuosum (Swartz) de Candolle. Except forD. tortuosum, the response increased positively with dose; the steepest slope occurred with an extract of cotton squares (flower buds). Almost all of the moths that landed on the extract dispenser also oviposited. Moths simulated by extracts from cotton squares exhibited a full array of behaviors (upwind flight, contact with the dispenser, examination of the cloth substrate with antennae, and oviposition) expected of gravid individuals seeking sites to propagate the species.This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation of its use by USDA.     

Larval exposure to oviposition deterrents alters subsequent oviposition behavior in generalist,Trichoplusia ni and specialist,Plutella xylostella moths

The present study was undertaken to determine the effects of larval feeding experience on subsequent oviposition behavior of the resulting moths. Larvae of the cabbage looper (Trichoplusia ni, Noctuidae) and the diamondback moth (Plutella xylostella, Plutellidae) were exposed to the phenylpropanoid allelochemical trans-anethole (at 100 ppm fw in artificial diet) or the limonoid allelochemical toosendanin (10 ppm sprayed on cabbage leaves). Both compounds had been shown to deter oviposition in naïve moths in previous choice tests. Moths developing from experienced larvae (both sexes) showed a decrease in oviposition deterrence response when given a choice between control and treated leaves, unlike naïve moths. This phenomenon, analogous to habituation to feeding deterrents in lepidopteran larva, occurred irrespective of duration of feeding on the deterrent compound. We also observed that F₁larvae resulting from experienced moths (previously exposed to toosendanin as larvae) grew as well on toosendanin-treated foliage as on control foliage. In contrast, growth of F₁larvae from naïve moths was significantly impaired by toosendanin. These results demonstrate that host-selection behavior in cabbage looper (a generalist) and diamondback moth (a specialist) may be shaped by feeding experience according to Hopkins' Host Selection Principle in addition to chemical legacy.     

Aqueous extracts from indigenous plants as oviposition deterrents forHeliothis virescens (F.)

In laboratory and field tests, water extracts from plant foliage deterred oviposition byHeliothis virescens (F.). A maximum reduction of 93% was attained in laboratory oviposition cages. When females were allowed free choice of treated or untreated tobacco plants in field cage and field tests, maximum reductions in oviposition were 71 and 83%, respectively.Lepidoptera: Noctuidae.Mention of a commercial or proprietary product does not constitute an endorsement by the USDA.     

Heliothis subflexa (Gn.) (Lepidoptera: Noctuidae): Demonstration of oviposition stimulant from groundcherry using novel bioassay

Methanol extracts of homogenized groundcherry,Physalis angulata, leaves increased egg deposition byHeliothis subflexa (Gn.) (HS) on treated tobacco plants (a nonhost) 8.5-fold over untreated controls. In doseresponse tests using whole-leaf washes of groundcherry leaves, the threshold of positive response vs. no response to the chemical stimulant was within one log dose unit when compared to the controls. This response was consistent whether the chemical was evaluated on plants in greenhouse-cage tests or in an olfactometer using pieces of broadcloth as the oviposition substrate. The olfactometer used allows year-round study of the behavioral effects of plant allelochemics on insect oviposition behavior in a controlled environment.This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation of its use by USDA.     

Oviposition stimulants of the mothEctomyelois ceratoniae: The effect of short-chain alcohols

Females of the mothEctomyelois ceratoniae prefer to oviposit on carobs,Ceratonia siliqua L., infested with the fungusPhomopsis sp. (Gothilf, 1964). An extract of the steam distillate of fungus-infested carobs was more effective in stimulating female moths to oviposit than was an extract of uninfested carobs. The extract of the fungus-infested carobs was found to be composed mainly of simple alcohols: ethanol (60%),n-propanol3 (15%), 2-propanol (2.5%), isobutanol (15%),n-butanol (2.5%), and isopentanol (5%). In addition, the extract contained a small proportion (1–2%) of unidentified compounds. All of the alcohols, with the exception of isopentanol, stimulated oviposition by female moths. However, the alcohols, when tested separately or as a mixture, were not as stimulating as the total extract. The unidentified compounds were less stimulating than the alcohols.     

Ovipositional response of tobacco budworm moths (Lepidoptera: Noctuidae) to cuticular labdanes and sucrose esters from the green leaves ofNicotiana glutinosa L. (Solanaceae)

Field plots of three accessions ofNicotiana glutinosa L. (Nicotiana species accessions 24, 24A, and 24B) at Oxford, North Carolina and Tifton, Georgia were heavily damaged by natural populations of tobacco budworms,Heliothis virescens (F.), during 1985–1989. Experiments in outdoor screen cages demonstrated that all accessions ofN. glutinosa were as prone to oviposition byH. virescens moths as was NC 2326, a commercial cultivar of flue-cured tobacco,N. tabacum L. However, in greenhouse experiments, tobacco budworm larvae did not survive or grow as well when placed on plants ofN. glutinosa as they did when placed on plants of NC 2326. Four labdane diterpenes (manool, 2-hydroxymanool, a mixture of sclareols, and labda-13-ene-8,15-diol [labdenediol]) and two sucrose ester fractions (2,3,4-tri-O-acyl-3-O-acetyl-sucrose [G-SE-I] and 2,3,4,-tri-O-acyl-sucrose [G-SE-II]) were isolated from green leaves of the three accessions ofN. glutinosa. These components were bioassayed for their effects on the ovipositional behavior of tobacco budworm moths using small screen cages in a greenhouse at Oxford, North Carolina. Labdenediol, manool, and both sucrose ester fractions stimulated tobacco budworm moths to oviposit on a tobacco budworm-resistant Tobacco Introduction, TI 1112 (PI 124166), when these materials were sprayed onto a leaf.This article reports the results of research only. Mention of a proprietary product does not constitute endorsement or recommendation for its use by USDA, ARS, or by the North Carolina Agricultural Research Service.     

Preferences of matedHeliothis virescens andH. subflexa females for host and nonhost volatiles in a flight tunnel

In flight-tunnel bioassays, mated femaleHeliothis subflexa (Gn.) moths demonstrated in dual-choice tests a significant preference for volatiles from an extract of their only known host,Physalis spp. (groundcherry). However,H. virescens (F.), a polyphagous species, responded positively by anemotaxis to extracts from susceptible tobacco, cotton,Desmodium tortuosum (host plants), and groundcherry, a nonhost.H. virescens females did not fly to volatiles emanating from an extract of a resistant tobacco cultivar.This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation of its use by USDA.     

Epicuticular Wax Chemicals in Zea mays Influence Oviposition in Ostrinia nubilalis

The chemical basis of oviposition elicitation in a generalist herbivore was determined by examination of oviposition responses in Ostrinia nubilalis to corn (Zea mays) chemicals in two-choice laboratory bioassays. A pentane extract of corn leaves stimulated oviposition and the activity persisted for three days, indicating that oviposition in O. nubilalis is elicited by low-volatility chemicals. Chemicals in the extract were fractionated by column chromatography on Florisil, using a sequence of solvents of increasing polarity. Bioassays of Florisil fractions indicated that the stimulants were eluted with nonpolar solvents. Positive bioassay results with an extract prepared by dipping corn leaves in pentane for 20 sec for extraction of leaf surface chemicals suggested that some of the active material was present in the leaf epicuticle. Gas chromatographic analyses and comparisons with retention times of standards suggested the presence of several n-alkanes in the dip extract. Five n-alkanes—hexacosane, heptacosane, octacosane, nonacosane, and tritriacontane—known to be present in the epicuticle of corn leaves were bioassayed, and all five elicited oviposition responses. These results suggest that oviposition elicitation in O. nubilalis is influenced by the presence of n-alkanes in the host plant epicuticle.     

Extracting Oviposition Stimulants for Carrot Fly from Host-Plant Leaves

Extracts of carrot foliage obtained with various extraction methods were compared for effectiveness in stimulating oviposition in the carrot fly. In choice assays, surrogate leaves treated with a hexane surface extract produced with a new microwave-assisted procedure were almost as acceptable as real host leaves. The high stimulatory activity of this extract was attributable to the raised solvent temperature, since cold hexane extracts were much less stimulatory. The microwave extract elicited about twice as much oviposition as the previously used dichloromethane surface extracts and the diethyl ether fraction of an extract that was obtained by brief immersion of leaves into water near its boiling point. The ovipositional responses to crude methanol and hot water extracts were weak because of the presence of yet unidentified polar deterrent compounds. Total extracts of ground foliage (vacuum distillation and extraction with liquid carbon dioxide) had no net stimulatory effect on oviposition.     

Evaluation of organic infusions and synthetic compounds mediating oviposition inAedes albopictus andAedes aegypti (Diptera: Culicidae)

Oviposition responses of gravidAedes albopictus andAe. aegypti were evaluated to natural organic infusions (hay infusion, larval rearing water, and field-collected larval water) as well as compounds isolated from hay infusion (3-methylindole, 4-methylphenol, 4-ethylphenol, indole, and phenol) known to elicit oviposition inCulex mosquitoes. In laboratory bioassays, significant oviposition responses were obtained fromAe. albopictus, but not fromAe. aegypti, to dilutions of hay infusion and field water. Oviposition responses of both species were moderate to the synthetic compounds tested in the laboratory (0.01–100 µg/liter). Only 3-methylindole (0.1 µg/liter) and 4-ethylphenol (1.0 µg/liter) elicited significantly more oviposition byAe. albopictus than did well water. Of the synthetic compounds tested withAe. aegypti, only phenol (1.0 µg/liter) and 4-ethylphenol (0.1 µg/liter) elicited significantly more oviposition than did well water. Significant repellency or oviposition deterrence for both species occurred in response to at least one high concentration of most of the compounds tested. In field cage evaluations, oviposition responses byAe. albopictus were strongest to larval water and field water, moderate to hay infusion and 3-methylindole (100 µg/liter), and low to well water. A mixture of five synthetic compounds mimicking hay infusion was no more effective than 3-methylindole alone. ForAe. aegypti, oviposition responses were greatest to larval water and least to 3-methylindole. In an olfactometer, gravid females of both species oriented more to field water than to well water and onlyAe. albopictus oriented more to larval water or hay infusion than well water. In general, gravidAe. albopictus responded more strongly to oviposition stimuli than didAe. aegypti.     

A Novel Bioassay for Yponomeuta cagnagellus Oviposition in Response to Extracts of Host and Nonhost Plant Surface Compounds

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.     

Influence of trichome exudates from species ofLycopersicon on oviposition behavior ofHeliothis zea (Boddie)

Cage experiments revealed that accessions of the wild tomato speciesLycopersicon hirsutum were preferred sites for oviposition byHeliothis zea. Hexane extracts from the leaves ofL. hirsutum were also preferred sites of oviposition in choice experiments among extracts from severalLycopersicon species. Extracts ofL. hirsutum were still biologically active several days after application, indicating that the phytochemical(s) involved are relatively stable and of low volatility. Gas Chromatographic analysis of leaf hexane extracts from 12 different accessions of theL. hirsutum complex and three tomato cultivars revealed substantial qualitative and quantitative variation in the chemical composition of these extracts. Comparison of these results with extract oviposition studies implicate a group of structurally related compounds as the active agents. Mass spectroscopy has tentatively identified these compounds as sesquiterpenes with the chemical formula C₁₅H₂₂O₂. These compounds are apparently synthesized and secreted from glandular trichomes on the leaf surface. These phytochemicals did not stimulate ovipositional behavior in females of the cabbage looper,Trichoplusia ni. The existence of genetic variation for the presence and amount of kairomones that serve as cues for insect orientation and oviposition could be utilized in a breeding program to develop tomato cultivars with genetically modified allelochemic profiles that would disrupt the sequential behavioral processes of insect host-plant selection.     

Contact stimulants fromHeliothis virescens that influence the behavior of females of the tachinid,Eucelatoria bryani

Factors acting at contact or close range affected the behavior associated with host seeking of females, but not of males, of the tachinid,Eucelatoria bryani Sabrosky. Females were arrested by components of larvae ofHeliothis virescens (F.) and by a dichloromethane extract of okra leaves. A hexane extract ofH. virescens frass and a chloroform-methanol extract ofH. virescens larvae were both active. In addition to chemicals, shape and size were involved because females were arrested by small inert objects the size ofH. virescens larvae and frass.In cooperation with the Louisiana Agricultural Experiment Station and the Texas Agricultural Experiment Station.Mention of a proprietary product does not constitute an endorsement by the USDA.     

Pyrrolizidine alkaloids as oviposition stimulants for the cinnabar moth,Tyria jacobaeae

In choice experiments with artificial leaves, we tested related pyrrolizidine alkaloids (PAs) for their stimulatory effects on the oviposition of the cinnabar moth, a specialist on the PA-containing plant Senecio jacobaea. The PAs from S. jacobaea that we tested stimulated oviposition. Monocrotaline also stimulated oviposition although this PA is not found in plants of the genus Senecio. The moths preferred ovipositing on filter paper with a PA mixture extracted from S. jacobaea to ovipositing on filter paper with single PAs. Senkirkine, heliotrine, and retrorsine did not stimulate oviposition. The nonactive retrorsine differs only in one OH group to the active senecionine, indicating that small structural differences alter the stimulatory activity of PAs. However, a PA mixture extracted from a nonhost plant, Senecio inaequidens, that consisted of 81% of the nonactive retrorsine did stimulate oviposition. Oviposition preferences between Senecio species seem to be determined by chemical compounds other than PAs.     

Oviposition response ofLobesia botrana females to long-chain free fatty acids and esters from its eggs

Avoidance of occupied ovisposition sites supposes that females perceive information related to their own progency. Fatty acids identified from egg extracts have been reevaluated using a different extraction method, and we have investigated the dose-dependent oviposition response of European grape vine moths (Lobesia botrana) to myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, methyl palmitate, methyl oleate, and ethyl palmitate; all except ethyl palmitate have been identified from eggs ofL. botrana. A methylene dichloride extract of eggs fromL. botrana revealed the presence of saturated free fatty acids (myristic, palmitic, and stearic) and unsaturated acids (palmitoleic, oleic, linoleic, and linolenic) in amounts ranging from 3.9 ng/egg equivalent for myristic acid to 30 ng/egg equivalent for palmitic and oleic acids. The extract also contained traces of methyl palmitate and methyl stearate. The greatest avoidance indexes were observed in response to palmitic, palmitoleic, and oleic acids. All the other compounds tested caused weaker responses. A reduction in the number of eggs laid was observed when moths were exposed to each of the esters applied at 0.3 µg per application spot. Reduction in eggs laid was also observed at a 10-fold higher dose of oleic acid. The present results confirm that general and simple molecules can be involved in the regulation of oviposition site selection and that they may participate in chemical marking of the eggs.     

Host-Plant Chemistry Influences Oviposition Choice of the Spicebush Swallowtail Butterfly

The spicebush swallowtail, Papilio troilus (L.), lays its eggs on plants in the family Lauraceae. Sassafras [Sassafras albidum (Nutt.) Nees], spicebush [Lindera benzoin (L.) Spreng.], redbay (Persea borbonia (L.)] and camphortree [Cinnamomum camphora (Nees) Eberm.] are four of its known host plants. In one-choice tests, free-flying spicebush swallowtail females laid eggs on chemical extracts of the leaves of each of these four hosts. In two-choice experiments, females always preferred to oviposit on an extract of sassafras compared to extracts of the other three hosts. It was shown for spicebush extract that this response was not due to oviposition experience. Previously we had identified one of the host plant chemicals acting as an oviposition stimulant in sassafras extract as 3-caffeoyl-muco-quinic acid (3-CmQA). Extracts of the other three hosts did not contain this compound. The addition of 3-CmQA alone to spicebush extract did not increase oviposition activity. It did, however, increase discrimination between hosts and nonhosts. When a fraction of sassafras extract containing 3-CmQA and other synergistic stimulants was added to spicebush extract, preference for sassafras extract was no longer recorded. These results show existing differences in oviposition chemistry among host plants of the spicebush swallowtail and how these differences can influence oviposition choice in bioassay experiments.     

Oviposition response of Chironomus tepperi to nitrogenous compounds and bioextracts in two-choice laboratory tests

Two-choice laboratory tests were used to investigate the oviposition response of Chironomus tepperi to a range of nitrogenous compounds and crude bioextracts. Responses to nitrogenous compounds varied in response to concentration. Ammonium nitrate did not influence oviposition at concentrations from 2 to 12 mg/liter. Hydroxylamine hydrochloride increased oviposition at 6 mg/liter, but had no effect at either 2 or 12 mg/liter. Sodium nitrate reduced oviposition at 2 mg/liter relative to the controls, but had no significant effect at 6 or 12 mg/liter. C. tepperi responded to many of the crude bioextracts, strongly avoiding oviposition in solutions containing homogenized chironomid larvae (1 final instar/100 ml; C. tepperi or Polypedilum nubiferum), and avoiding solutions conditioned by conspecificlarvae at concentrations down to the equivalent of 1 final instar/100 ml over 24 hr. Homogenates of adult conspecifics had no effect on oviposition site selection. Homogenates of larval Culex annulirostris (Culicidae) deterred oviposition, but only at high concentrations (3 final instars/100 ml). Our results demonstrate that chemical cues from larval populations deter oviposition by C. tepperi females searching for newly flooded habitats where larval competition will be minimized.     

Identification and bioassay of kairomones forHelicoverpa zea

Hexane extracts of leaves of 307 accessions from 73 host plant species ofHelicoverpa zea were analyzed by gas chromatography (GC) and used forH. zea oviposition and neonate larvae orientation bioassays. The gas chromatographic (GC) retention times of compounds statistically associated with behavioral activity were identified by correlation of GC peak area with oviposition and larval orientation preferences. Although taxonomically diverse in their origin, compounds for study were purified from extracts of species of the genusLycopersicon, due to their relative abundance. The structures of eight long-chain alkanes associated with oviposition preference were assigned by mass spectrometry, and the structures of five similarly associated organic acids and a terpenoid alkene were identified by¹H and¹³C nuclear magnetic resonance spectroscopy. The structures of a number of other phytochemicals from the plant leaves were identified for comparative purposes, including a previously unknown terpene, 7-epizingiberene. Bioassays were performed on the isolated acids and on the alkane wax fractions of severalLycopersicon species, and significant differences were found in oviposition stimulation for both classes of compounds. Of the hundreds of compounds found in the extracts, none were observed to act as oviposition deterrents. The results of these bioassays may be useful in explaining the broad host range ofH. zea, as well as the process and evolution of host plant selection for oviposition.