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.     

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.     

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.     

Sex pheromone specificity inHeliothis

Electric grid traps baited withHeliothis subflexa (Guenée),H. virescens (F.), orH. zea (Boddie) females captured conspecific males with few exceptions.Heliothis subflexa females reduced the attraction ofH. virescens andH. zea males when used as bait simultaneously with females of either of these two species. Backcrosses were made withH. virescens males and female hybrids from a cross betweenH. subflexa females andH. virescens males. The backcross (BC) females andH. virescens females attracted approximately equal numbers ofH. virescens males in field traps. BC males released in field cages were attracted toH. virescens females and to the synthetic pheromone ofH. virescens. When laboratory-reared maleH. virescens, BC males,H. subflexa males, and F₁ hybrid males were exposed to the synthetic pheromone ofH. virescens in Plexiglas wind tunnels,H. virescens males and BC males responded to the pheromone, butH. subflexa and F₁ hybrid males did not. The peak activity of bothH. subflexa andH. zea males occurred approx. 4 hr after sunset. MaleH. zea were active throughout most of the night; maleH. virescens were most active approx. 6 hr after sunset.This paper reports the results of research only. Mention of a pesticide or of a commercial or proprietary product in this paper does not constitute a recommendation for use by the U.S. Department of Agriculture nor does it imply registration under FIFRA as amended.Lepidoptera: Noctuidae.     

Production of pheromone by hairpencil glands of males obtained from interspecific hybridization betweenHeliothis virescens andH. subflexa (Lepidoptera: Noctuidae)

Pheromone produced by the hairpencil glands of interspecific hybrid- and backcross-generation males from crosses betweenHeliothis virescens (F.) withH. subflexa (Gn.) was studied. Males of reciprocal F₁ hybrids, all of which had hairpencil glands morphologically similar to those ofH. virescens, produced neither the same pheromone blend nor amounts of pheromone that were produced by males ofH. virescens. Instead, these hybrid males produced pheromone that was quantitatively and qualitatively similar to that produced byH. subflexa. Hairpencil gland extracts from males obtained from backcrossing F₁ females of either cross to males ofH. subflexa were the same as those ofH. subflexa. However, extracts from backcross males of crosses between F₁ females andH. virescens were variable. Some extracts from these backcross males were like those ofH. virescens while others were either likeH. subflexa or were intermediate between those of the parent species. These results showed that the production of pheromone by the hairpencil glands of hybrid and backcross males is under the dominant regulation of autosomal genes of theH. subflexa genome.     

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.     

Hairpencil Volatiles Influence Interspecific Courtship and Mating Between Two Related Moth Species

Reproductive isolation between sympatric, closely related species can be accomplished through a variety of pre-zygotic isolating mechanisms, including courtship-signaling behavior that involves pheromones. In the moths Heliothis virescens and H. subflexa, males display abdominal hairpencils (HP), which release volatile chemicals during courtship. In this study, we demonstrated that HP volatiles released by male H. subflexa function to improve mating success with conspecific females. Interspecific mating experiments were conducted to determine any influence of HP volatiles on species isolation. Female H. virescens and H. subflexa were observed during courtship with males of the other species, following either sham-operation or ablation of HPs, both with and without concurrent presentation of HP volatiles. Mating success was improved by co-presentation of HP extract from males of the same species during courtship. Ablation of HPs improved mating between H. subflexa females and H. virescens males. During interspecific matings, male H. virescens attempted copulation less frequently in the presence of H. virescens HP extract, though H. subflexa males were not affected by the presence of H. subflexa HP extract. This suggests that HP volatiles produced by males of each species may inhibit mating between species through effects on males (H. virescens) and females (H. subflexa).     

Kairomones and their use for management of entomophagous insects

Volatile chemicals emanating from an excretion (apparently meconium) and abdominal tips of femaleHeliothis zea (Boddie) moths mediated increased rates of parasitization ofH. zea eggs byTrichogramma pretiosum Riley. A blend of synthetic chemicals, consisting of hexadecanal, (Z)-7-hexadecenal, (Z)-9-hexadecenal, and (Z)-11-hexadecenal, which has been identified as the sex pheromone of and from the abdominal tip of femaleH. zea moths, also increased rates of parasitization ofH. zea eggs byT. pretiosum in greenhouse experiments. In addition, parasitization ofH. zea eggs by wildTrichogramma spp., in field plots of cotton,Gossypium hirsutum L., treated with a similar blend of chemicals, in Conrel fibers, was more than double that in untreated plots.Hymenoptera: Trichogrammatidae.Lepidoptera: Noctuidae.In cooperation with the University of Georgea College of Agriculture Experiment Stations, Coastal Plain Station, Tifton, Georgia 31793.Mention of a proprietary product does not constitute endorsement by the USDA.     

Host selection byHylemya antiqua laboratory bioassay and methods of obtaining host volatiles

An oviposition bioassay for the onion maggot,Hylemya antiqua (Meigen), is described in which females, in response to volatile stimulants, oviposit through small apertures onto moistened filter paper. Onion volatiles that act as attractants and oviposition stimulants were captured in Porapak Q from air passed over chopped onions in glass chambers. Pentane extracts from odor-impregnated Porapak Q induced 30–50% of the oviposition that occurred in response to 15-g onion-slice stimuli. Extracts presented in pentane on waxed dental cotton wicks induced more constant oviposition over a 3-day period than extracts on unwaxed wicks. Extract of the Porapak Q-captured volatiles from bulbs of fresh, actively growing onions elicited a much stronger response than did stem and leaf extracts from the same onions. The bioassay techniques and chemical procedures developed in this study could be used in chemical isolation programs for host attractants or oviposition stimulants forH. antiqua or similar species.Diptera:Anthomyiidae.Research supported by National Research Council of Canada, Grant Nos. A3881 and A3706.     

Kairomones and their use for management of entomophagous insects: III. Stimulation ofTrichogramma achaeae,T. pretiosum,andMicroplitis croceipes with host-seeking stimuli at time of release to improve their efficiency

Frass from larvae of the corn earworm,Heliothis zea (Boddie) and scales fromH. zea moths (that are known to contain the host-seeking stimulus, tricosane) stimulate and orient host-seeking activity in femaleMicroplitis croceipes (Cresson), a larval parasite ofH. zea, andTrichogramma spp., egg parasites ofH. zea. When larval frass, moth scales, and tricosane were used as sign stimuli (releasers) forM. croceipes, T. pretiosum (Riley), andT. achaeae Nagaraji and Nagarkatti, respectively, at time of their release from laboratory containers, parasite performance improved, resulting in significantly increased rates of parasitization over that of unstimulated parasites. Stimulation ofM. croceipes with larval frass had an overriding effect on this parasite's innate tendency to disperse upon release, thereby increasing the numbers remaining and prolonging their retention in the target area. Supplying the appropriate host-seeking stimuli to these 3 hymenopterous parasites ofH. zea at time of their release to improve their efficiency greatly increases the probability of their effective utilization in pest management systems.     

Flight response ofHeliothis subflexa (Gn.) females (Lepidoptera: Noctuidae) to an attractant from groundcherry,Physalis angulata L.

Mated femaleHeliothis subflexa (Gn.) (HS) moths 1–7 days old responded positively in a Plexiglas flight tunnel to an attractant extracted with methanol from fresh whole-leaf washes of groundcherry,Physalis angulata L. Response to the groundcherry extract, as indicated by plume-tracking (i.e., upwind flight toward the odor source) and contact with the chemical dispenser did not change significantly during the first 5 hr of scotophase. Overall, ca. 50% of the responding moths also landed on the chemical dispenser; ca. 50% of the moths that landed also deposited eggs. There were no significant differences in the behavioral responses of females mating only once and those that had mated two or more times. Virgin females and male moths were significantly less responsive to the groundcherry attractant than mated females. The flight tunnel bioassay described provides an excellent system for evaluating plant allelochemics associated with host-plant selection.This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation for its use by USDA.     

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.     

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.     

Behavior ofHeliothis virescens (F.) in presence of oviposition deterrents from elderberry

Extracts prepared from elderberry,Sambucus simpsonii Rehd., leaves with either acetone, dichloromethane, distilled water, ethanol, hexane, or methanol deterred oviposition byHeliothis virescens (F.) on treated substrates in the laboratory. Doses of the aqueous extract equivalent to as little as 0.8 mg of leaves/cm2 of oviposition substrate were effective in reducing egg deposition. There was no significant difference in the mean number of landings on extract-treated and untreated surfaces. When either the antennae, proboscis, or the metathoracic legs were removed from female moths, there was no significant effect on oviposition on paper towels treated with elderberry leaf-water extract in laboratory bioassays, but in field cages, moths without these appendages deposited significantly fewer eggs on treated leaves of tobacco plants, a preferred host. There was no evidence that elderberry leaf-water extract affected mating byH. virescens.Lepidoptera: Noctuidae.Mention of a commercial or proprietary product does not constitute an endorsement by the USDA.     

Terminal steps in pheromone biosynthesis byHeliothis virescens andH. zea

In vivo application to the sex pheromone gland ofHeliothis Virescens andH. Zea of large quantities of alcohols normally present in small amounts resulted in the preferential conversion of the alcohols to the corresponding pheromonal aldehydes. Amounts of the minor component aldehydes were increased up to 15-fold by selectively applying large quantities of the alcohol precursors. Using this technique, we have inducedH. virescens to convert bombykol, the sex pheromone of the silkworm, to the corresponding aldehyde, bombykal, and have induced femaleH. zea to produce the same sex pheromone components used byH. virescens by applying tetradecanol and (Z)-9-tetradecenol to the surface of the gland. Further, treatedH. zea females were attractive toH. virescens males and caused males to attempt interspecific copulation repeatedly. We have also found that the enyzme involved in this conversion is dependent on the presence of molecular oxygen, indicating that a nonspecific alcohol oxidase is responsible for the terminal biosynthetic step in pheromone production by bothH. virescens andH. zea.Mention of a commercial product or proprietary does not constitute endorsement by either the University of Guelph or U.S.D.A.     

Analyses of an egg recognition kairomone ofTelenomus heliothidis (Hymenoptera: Scelionidae) Isolation and host function

Two large proteins (>330 K) present in the accessory gland ofHeliothis virescens F. (Lepidoptera: Noctuidae) serve as an egg recognition kairomone for the parasitoidTelenomus heliothidis Ashmead (Hymenoptera: Scelionidae). Glass beads coated with the isolated proteins were examined and probed by the parasitoid while uncoated glass beads were not. Electrophoretic analysis of the contents of theH. virescens accessory gland suggests that the gland's contents serve as an egg adhesive. Comparison of the gland proteins revealed that theT. heliothidis hostHeliothis zea (Boddie) (Lepidoptera: Noctuidae) accessory gland contained the kairomonally active proteins while the nonhostSpodoptera frugiperda (Lepidoptera: Noctuidae) did not.     

Impact of Caterpillar Increased Feeding Rates on Reduction of Bt Susceptibility

The use of insect-resistant transgenic crops producing Bacillus thuringiensis protein Cry toxins (Bt) to control caterpillars is wide-spread. Development of a mechanism to prevent Bt from reaching its target site in the digestive system could result in Bt resistance and resistance to other insecticides active per os. Increased feeding rates by increasing temperature in tobacco budworms, Chloridea virescens, and bollworms, Helicoverpa zea, decreased Bt Cry1Ac susceptibility and mortality. The same was found in C. virescens for Bollgard II plant extract containing Bt Cry1Ac and Cry2Ab2 toxins. Furthermore, H. zea from the same inbred laboratory colony that fed faster independent of temperature manipulation were less susceptible to Bt intoxication. A laboratory derived C. virescens Bt resistant strain demonstrated a higher feeding rate on non-Bt artificial diet than the parental, Bt susceptible strain. A laboratory-reared Bt resistant fall armyworm, Spodoptera frugiperda, strain also fed faster on non-Bt diet compared to Bt susceptible caterpillars of the same species, both originally collected from corn. The studies in toto and the literature reviewed support the hypothesis that increased feeding rate is a behavioral mechanism for reducing caterpillar susceptibility to Bt. Its possible role in resistance needs further study.     

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.     

Sex pheromone chemistry of the female tobacco budworm moth,Heliothis virescens

(Z)-11 -Hexadecenal (77–91%), (Z)-7-hexadecenal (0.1–2%), (Z)-9-hexadecenal (0.3–2%), hexadecanal (3–19%), (Z)-11-hexadecen-1-ol (1–5%), tetradecanal (1–3%), and (Z)-9-tetradecenal (1–3%) were identified from the heptane washes of the ovipositor of femaleHeliothis virescens (F.) females. In field bioassays, a 152-g mixture of these seven compounds deployed in an insect trap exceeded the attractiveness of 4 virgin femaleH. virescens for males and was 5–6 times more attractive than a mixture of (Z)-1 1-hexadecenal and (Z)-9-tetradecenal (virelure) that was previously reported as the sex pheromone of the species. Four of the seven compounds produced byH. virescens females are also produced byH. zea (Boddie). Specificity of pheromonal signals among the two species is ostensibly dependent upon one or more of the three additional compounds [tetradecanal, (Z)-9-tetradecenal, and (Z)-1 1-hexadecen-1-ol] produced by femaleH. virescens.     

Taste Sensitivity of Insect Herbivores to Deterrents is Greater in Specialists Than in Generalists: A Behavioral Test of the Hypothesis with Two Closely Related Caterpillars

Sensitivity of caterpillars of Heliothis virescens, a generalist, and H. subflexa, a specialist, to eight different plant secondary compounds was examined behaviorally. The compounds were nicotine hydrogen tartrate, hordenine, caffeine, sinigrin, linamarin, arbutin, chlorogenic acid, and salicin. All compounds deterred feeding, at least at the higher concentrations, but the generalist was less affected than the specialist. Thus the hypothesis that specialists have greater sensitivity to deterrents than generalists was supported. In most cases deterrence occurred on first encounter, indicating that the response was sensory; in some cases short-term postingestive effects also appeared to play a role. The larger quantities of deterrent-containing food ingested by H. virescens sometimes resulted in measurable postingestive effects during the second control test. This did not occur in H. subflexa, which more commonly rejected the deterrent-containing food on first contact. The contrast between the species is discussed in relation to tradeoffs involved in different diet breadths.