Insect antifeedant activity associated with compounds isolated from species ofLonchocarpus andTephrosia

The antifeedant activity of a series of 21 chalcones, flavanes, and flavanones isolated from the generaLonchocarpus andTephrosia (Leguminosae) was assessed by behavioral and electrophysiological bioassays against larvae ofSpodoptera littoralis andS. exempta. The antifeedant activity is related to the molecular structure of the compounds, and possible modes of interaction with the insect taste receptors are discussed.     

Insect antifeedant activity of clerodane diterpenoids against larvae ofSpodoptera Littoralis (Boisd.) (Lepidoptera)

Antifeedant activities of nine clerodane diterpenoids, isolated in this laboratory from different species ofAjuga plants, have been studied against larvae of Egyptian cotton leafwormSpodoptera littoralis (Boisd.) (Lepidoptera) by application of the leaf disk method. Evaluation of activity was carried out by calculating, at different time intervals, the feeding ratio (FR) from the relationship between the consumed areas of treated disks (CTD) and control disks (CCD); for comparison purposes a FR₅₀ defined as the FR at a CCD of 50% was established. Some compounds exhibited activity at a 0.01 g/cm² dose (0.3 ppm). Structure-activity relationships are discussed     

Reimer-Tiemann adducts as potential insect antifeedant agents Reviewing the structure-activity relationship theory of the antifeedant,warburganal

Structure-activity relationships of naturally occurring enedials with antifeedant activity againstSpodoptera species have been extended via the synthesis and bioassay of a series of Reimer-Tiemann adducts. The activities attributed to the different chemical structures of these and other analogs interacting with the chemoreceptor site have been observed; a three-pronged mode of substrate binding via aromatic pyrrole formation, Michael addition of free sulfhydryl moieties, and van der Waals interactions of the A ring has been postulated to account for these observations.Names of products in this paper are included for the benefit of the reader and do not imply endorsement or preferential treatment by USDA.     

A new method for fast isolation of insect antifeedant compounds from complex mixtures

Bioassay-guided isolation of bioactive natural substances requires monitoring of all fractionation and purification steps using a bioassay system. This is often a long and tedious process, especially with insect feeding bioassays. We report here a new method, based on the principle of bioautography, for a quick isolation of insect antifeedant compounds. TLC plates, after development, are coated with a thin layer of artificial diet and fed toSpodoptera litura larvae. The location of uneaten areas is then compared with theR _fvalues of the TLC spots, in order to determine rapidly the active fractions. This methods allows for a very fast determination of the most active antifeedant compounds in a complex mixture and considerably speeds up the isolation process. This new method was successfully applied in the study of antifeedant activity of several plant samples, and results are presented here for a model plant,Skimmia japonica (Rutaceae). Using this new method, the compounds responsible for the feeding-deterrent activity, three furanocoumarins (bergapten, xanthotoxin, and oxypeucedanin), were quickly and efficiently identified.     

Effect of Trichilia americana Extract on Feeding Behavior of Asian Armyworm, Spodoptera litura

We investigated the desensitization of Asian armyworms, Spodoptera litura, to a crude methanolic extract of wood Trichilia americana. Following repeated exposures to the extract over four days, desensitization was observed under no-choice conditions at low extract concentration (0.5 g/cm²). No desensitization was observed under choice conditions or at a higher concentration (5.0 g/cm²). Repeated exposures of larvae to the extract over 7.5 hr showed that larvae increasingly tolerated the extract due to hunger, rather than showing true desensitization. When larvae were exposed to the extract in test arenas of different sizes, larvae showed reduced feeding and desensitization as arena size increased. Finally, treatment of whole cabbage plants with extract resulted in larvae abandoning the plant. The crude extract was effective at protecting the plants, although desensitization did not occur, because the larvae moved away from the plant.     

Camphor from juvenile white spruce as an antifeedant for snowshoe hares

One theory in plant antiherbivore defense predicts that slow growing late succession plants like white spruce (Picea glauca) make large investments in antiherbivore defenses. Juvenile stages of white spruce in the Yukon, Canada, are rarely browsed by snowshoe hares (Lepus americanus), an abundant herbivore, but mature spruce is a highly preferred food. The hexane-soluble fractions of the methanol extracts from juvenile and mature white spruce contain camphor and bomyl acetate. There is four times as much camphor in juvenile spruce as in mature spruce from GC analysis. Plant extracts were added to rabbit chow. Pairs of extracts were offered to hares in choice tests. These tests demonstrated that camphor in the juvenile spruce extracts deterred feeding. Bornyl acetate did not have a clear antifeeding effect.On leave from the Department of Chemistry, University of South Pacific, Fiji.     

Behavioral and electrophysiological study of antifeedant mechanisms associated with polyhydroxy alkaloids

Eleven polyhydroxy alkaloids of plant origin were tested for anti-feedant effects against larvae of the lepidopteransSpodoptera littoralis, Spodoptera frugiperda, Heliothis virescens, andHelicoverpa armigera. Data from behavioral and electrophysiological investigations were correlated to reveal information on the mode of action of the antifeedants. The pyrrolidine DMDP was an effective antifeedant for all four species, whereas the piperidines fagomine and XZ-1 and the pyrrolizidine alexine were all ineffective as antifeedants. The activity of the pyrrolidines CYB-3 and DAB-1, the piperidines DNJ, DMJ, and BR1, and the bicyclic octahydroindolizine castanospermine varied among species. The investigation focuses on the structural similarities between some of the alkaloids and some common phagostimulatory sugars and illustrates a neural interaction involving the neurons that are differentially responsive to alkaloids and sugars. InS. littoralis, the neurons responding specifically to the alkaloids DMDP, DAB-1, and castanospermine and to the sugars fructose, sucrose, and glucose are more active when the compounds are applied singly than when an alkaloid and a sugar are applied together. The implications for the occurrence and functioning of different sugar receptor sites are discussed.     

Insect olfactory receptor responses to components of pheromone blends

Multicomponent pheromone systems are common in many insect species. As our knowledge about the number of different chemical compounds actually involved in a particular communication system increases, so too does the need for an efficient neural mechanism for the encoding of behaviorially relevant odor compounds. Here we consider the electrical activity of olfactory receptor neurons in a subset of the individual pheromone-sensitive sensilla on the antennae of male cabbage looper moths (Trichoplupia ni). Responses to single- and multiple-component stimuli, drawn from seven behaviorally active compounds, were obtained at several different intensities. Some blends elicited electrical responses which were not readily predicted from a knowledge of the receptor neuron's response to individual components.     

Detrimental effects ofCinchona leaf alkaloids on larvae of the polyphagous insectSpodoptera exigua

YoungCinchona ledgeriana plants contain two types of alkaloid: indole alkaloids in the leaves and quinoline alkaloids in the root. FromCinchona leaves, a crude alkaloid extract was made, containing the cinchophylline type of indole alkaloids and a small amount of 5-methoxytryptamine. The leaf alkaloid extract exerted a strong detrimental effect on the growth of larvae of the polyphagous beet armyworm,Spodoptera exigua (Lepidoptera). Feeding of larvae on an artificial diet containing the leaf alkaloids at the same concentrations as those found in the plant resulted in significant growth reduction, retardation in development, and mortality of the larvae. Cinchophyllines are composed of 5-methoxytryptamine coupled to a corynantheal unit. When incorporated into the artificial diet, 5-methoxytryptamine alone had no effect on the 5.exigua larvae. Corynantheal, however, had a strong detrimental effect on growth of the larvae, its effect being comparable to that of the leaf alkaloid extract. In contrast to the indole-type leaf alkaloids, the quinolinetypeCinchona root alkaloids did not affect growth and development of the larvae. These results suggest that the indole-type alkaloids, which inCinchona plants are present at the highest concentrations in the young, vulnerable leaflets, are involved in the chemical defense of the plant against herbivorous insects.     

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.     

Response of parasitoidEucelatoria bryani to selected plant material in an olfactometer

The response of the parasitoidEucelatoria bryani Sabrosky to fresh plant material and several plant extracts was examined in an olfactometer. The plants tested can serve as a food source for hosts (Heliothis spp.) of the parasitoid. Females responded positively to 13 of the 19 fresh plant tissues tested, and all of the extracts tested. Males responded to only two of the fresh plant tissues and to none of the extracts. The significance of plant variety, morphology, and phenology on the host habitat-selection behavior of this parasitoid is discussed.     

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.     

Dodecyl acetate,a second pheromone component of the cabbage looper moth,Trichoplusia ni

Dodecyl acetate was identified as a second component of the sex pheromone ofTrichoplusia ni (Hübner). Dodecyl acetate comprised 9.6% by weight of the total pheromone [(Z)-7-dodecenyl acetate plus dodecyl acetate] extracted from glands and 7.3% by weight of the total pheromone evaporated from the surfaces of glands. Dodecyl acetate appears to function as a short-range pheromone component. Evaporation at female release rates of a 1090 mixture of dodecyl acetate and (Z)-7-dodecenyl acetate in the field caused a significantly greater percentage of males to land on the pheromone source, increased significantly the time they spent on the source, but decreased significantly the time they spent searching for the source when within 50 cm, as compared to (Z)-7-dodecenyl acetate alone.Lepidoptera: Noctuidae.     

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.     

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.     

Analysis of the Insect OS-D-Like Gene Family

Insect OS-D-like proteins, also known as chemosensory (CSP) or sensory appendage proteins (SAP), are broadly expressed in various insect tissues, where they are thought to bind short to medium chain length fatty acids and their derivatives. Although their specific function remains uncertain, OS-D-like members have been isolated from sensory organs (including the sensillum lymph in some cases), and a role in olfaction similar to that of the insect odorant binding proteins (OBP) has been suggested for some. We have identified 15 new OS-D-like sequences: four from cDNA clones described herein and 11 from sequence databases. The os-d-like genes from the Anopheles gambiae, Apis mellifera, Drosophila melanogaster, and Drosophila pseudoobscura genomes typically have single, small introns with a conserved splice site. Together with all family members entered on GenBank, a total of 70 OS-D-like proteins, representing the insect orders Diptera, Dictyoptera, Hymenoptera, Lepidoptera, Orthoptera, and Phasmatodea, were analyzed. A neighbor joining distance phenogram identified several protein similarity classes that were characterized by highly conserved sequence motifs, including (A) N-terminal YTTKYDN(V/I)(N/D)(L/V)DEIL, (B) central DGKELKXX(I/L)PDAL, and (C) C-terminal KYDP. In contrast, three similarity classes were characterized by their diversion from these conserved motifs. The functional importance of conserved amino acid residues is discussed in relation to the crystal and NMR structures of MbraCSPA6.     

Responses from sensilla on antennae of maleHeliothis zea to its major pheromone component and two analogs

For some species, chemical analogs have been identified that can substitute for the major pheromone component and reduce pheromone-trap captures in atmospheric permeation experiments. The ability to substitute these analogs for the major pheromone component in field tests raises the question: Do the same set of olfactory receptor neurons on the insects' antennae respond to both the major component and the effective structural analogs? To investigate this question, extracellular responses were recorded from single sensilla on the antennae of maleHeliothis zea (Boddie) (Lepidoptera: Noctuidae) to stimulation with increasing doses of (Z)-11-hexadecenal, the major pheromone component, (Z)-9-tetradecen-1-ol formate, and (Z)-1,12-heptadecadiene. Both (Z)-9-tetradecen-1-ol formate and (Z)-1,12-heptadecadiene can substitute for (Z)-11-hexadecenal in trap disruption experiments and affect the sexual behavior of males, although neither has been identified in the volatiles released by calling females. All three compounds elicited responses from the same class of pheromone-sensitive receptor neurons over a range of stimulus concentrations. At equivalent stimulus concentrations, however, (Z)-11-hexadecenal elicited about a 10-fold greater response than (Z)-1,12-heptadecadiene. (Z)-9-Tetradecen-1-ol formate elicited a response slightly higher than, but not significantly different from, the response to (Z)-1,12-heptadecadiene.     

Behavioral responses of maleHeliothis zea moths in sustained-flight tunnel to combinations of 4 compounds identified from female sex pheromone gland

Each of the four compounds that have been identified from sex pheromone glands ofHeliothis zea female moths was examined for its ability to elicit sexual responses from male moths in a flight tunnel. Males flew upwind to (Z)-11-hexadecenal alone, but greater levels of behavioral activity were evoked with the addition of (Z)-9-hexadecenal to the treatment. Addition of hexadecanal or (Z)-7-hexadecenal to the initial two components had no effect in raising the behavioral response of the males in the flight tunnel whether added singularly at both the normal gland-emission ratio or at varying ratios or in combination at the normal ratio. Live, calling females elicited levels of sexual activity from males not significantly different from that elicited by the mixture of (Z)-11- and (Z)-9-hexadecenal on cotton wicks.Lepidoptera: Noctuidae.     

Evidence for presence and nature of female sex pheromone ofBrithys crini fabricius (Lepidoptera: Noctuidae)

The existence of a female sex pheromone of the noctuid mothBrithys crini Fabricius was confirmed in both laboratory bioassay and field tests. Crude extracts and airborne volatiles from females were analyzed by gas chromatography and mass spectrometry and the data compared with authentic compounds. The primary sex pheromonal compound was Z11-16: Ald. Scanning electron microscopy showed that the external surface of the sex pheromone gland was covered with folds that might increase the sex pheromone evaporation area.     

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.