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.     

Effects of pheromone components and their degradation products on the response ofHeliothis spp. to traps

None of the isolated degradation products of (Z)-11-hexadecenal [(Z)-11-HDAL] affected the catches of either tobacco budworm [Heliothis virescens (F.)] or bollworm [H. zea (Boddie)] moths when dispensed with pheromone from cotton dental rolls in cone traps. Also, none of the degradation products of (Z)-9-tetradecenal [(Z)-9-TDAL] had an effect on trap catches of tobacco budworm moths. Two of the three chemicals that have previously been identified in ovipositor washes of tobacco budworms but that are absent in those of bollworms caused a reduction in capture of bollworms: (Z)-9-TDAL (1.0 g/trap) caused a 96% reduction in trap catch and (Z)-11-hexadecen-1-ol (20.0 g/trap) caused a similar reduction. Tetradencenal (40 g/trap) had no effect on trap catch.In cooperation with the Texas Agricultural Experiment Station, Texas A&M University, College Station, Texas 77843.This paper reports the results of research only. Mention of a pesticide in this paper does not constitute a recommendation for use by the USDA nor does it imply registration under FIFRA as amended. Also, mention of a commercial or proprietary product in this paper does not constitute an endorsement of this product by the USDA.     

Influence ofHeliothis virescens sex pheromone dispensers on captures ofH. zea males in pheromone traps relative to distance and wind direction

Evaluations conducted by placingHeliothis virescens (F.) sex pheromone (virelure) dispensers at different distances in the predominant downwind and upwind directions fromHeliothis zea (Boddie) pheromone traps indicated that reductions inH. zea male captures were greatest relative to distance when theH. zea traps were located downwind from the virelure dispensers than when the traps were located upwind. When operating traps for both species at the same site, the influence of virelure dispensers on captures inH. zea pheromone traps would be minimized by placing theH. zea traps upwind of theH. virescens traps and, if wind direction is variable, the traps should be spaced at least 75 m apart.Lepidoptera: Noctuidae.In cooperation with the Texas Agricultural Experiment Station, Texas A&M University, College Station, Texas 77843. This paper reports the results of research only. Mention of a commercial or proprietary product in this paper does not constitute an endorsement of this product by USDA.     

Identification of a sex pheromone ofHeliothis subflexa (GN.) (Lepidoptera: Noctuidae) and field trapping studies using different blends of components

Eight compounds were isolated from the sex pheromone gland ofHeliothis subflexa (Gn.) and identified as hexadecanal, (Z)-9-hexadecenal, (Z)-11-hexadecenal, (Z)-7-hexadecen-1-ol acetate, (Z)-9-hexadecen-1-ol acetate, (Z)-11-hexadecen-1-ol acetate, (Z)-9-hexadecen-1-ol, and (Z)-11-hexadecen-1-ol. Although the whole blend was found to be an effective male attractant, the deletion of alcohols from the blend increased trap captures considerably. Further, although the binary mixture of (Z)-9-hexadecenal and (Z)-11-hexadecenal caught some maleH. subflexa, significant increases in captures were noted when the three acetate components were included in the blend.Mention of a commercial or proprietary product in this paper does not constitute an endorsement of that product by the USDA or the State of Florida.     

Properties of cuticular oxidases used for sex pheromone biosynthesis byHeliothis zea

Biosynthesis of the aldehydic sex pheromone components released by females ofHeliothis zea was found to be catalyzed by primary alcohol oxidases residing in the cuticle that covers the glands. Activity, as indicated by conversion of primary alcohol to aldehyde, was as high in cell-free cuticle as it was in intact pheromone glands. Studies indicated that some activity was associated with the surface of the epicuticle and could be removed, into buffer, by sonication. However, the majority of activity lies within the inner epicuticle and exo- and endocuticular layers. The oxidase was not functional in pharate pupae that did not have mature adult cuticle but became functional just prior to adult emergence. The enzyme in individual glands was saturated at alcohol concentrations above 100 n. moles. Nonionic detergents did not affect the activity of the oxidase in the cuticle but treatment with either 7 M urea or 1% SDS resulted in total loss of activity. Studies on the effect of pH indicated an optimum at 6.4; however, activity was high throughout the range of 5–9. The oxidase was functional in both dichloromethane and hexane, suggesting that this enzyme system may have applications for organic synthesis of aldehydes.     

EFFECT OF PBAN ON PHEROMONE PRODUCTION BY MATED <Emphasis Type="Italic">Heliothis virescens</Emphasis> AND <Emphasis Type="Italic">Heliothis subflexa</Emphasis> FEMALES

Mated female Heliothis virescens and H. subflexa were induced to produce sex pheromone during the photophase by injection of pheromone biosynthesis activating neuropeptide (PBAN). When injected with 1 pmol Hez-PBAN, the total amount of pheromone that could be extracted from glands of mated females during the photophase was similar to that extracted from virgin females in the scotophase. The PBAN-induced profile of pheromone components was compared between mated, PBAN-injected females and virgin females during spring and fall. Virgin females exhibited some differences in the relative composition of the pheromone blend between spring and fall, but no such temporal differences were detected in PBAN-injected, mated females. Because the temporal variation in pheromone blend composition was greater for virgin females than for PBAN-injected females, PBAN can be used to determine a females native pheromone phenotype. This procedure has the advantages that pheromone glands can be extracted during the photophase, from mated females that have already oviposited.     

Attractiveness of tobacco budworm females altered by oral chemosterilants and dietary additives

FemaleHeliothis virescens (F.) moths reared from larvae on diet treated (0.1%) with experimental chemosterilants or the dietary additive, DL-leucine, were used as bait in sex lure traps in field cages when they were 2–4 nights old. Catches of untreated released males were used to determine relative attractiveness of the chemically treated females. The catch (indicating quantity or production frequency of pheromone) was significantly increased whendl-leucine had been fed in the larval diet, and sulfanilamide caused a slight increase in female attractiveness. The catch of males was significantly reduced when either reserpine or quercitin had been added to the diet. The other chemicals, bisdicumarol, 2,4-dichlorophenoxyacetic acid, -sitosterol, and dihydrocholesterol, did not significantly affect the catch.This paper reports the results of research only. Mention of a pesticide in this paper does not constitute recommendation of use by the U.S. Department of Agriculture nor does it imply registration under FIFRA as amended.In cooperation with the Texas Agricultural Experiment Station, Texas A&M University, College Station 77843.The assistance of C.T. Perez and R.J. Guerra, of this laboratory, during field tests is gratefully acknowledged.     

Sex pheromone source location by garter snakes:

Male plains garter snakes,Thamnophis radix, tested in a 240-cm-long arena can detect directional information from a female pheromone trail only when the female is allowed to push against pegs while laying the trail. The female's normal locomotor activity apparently deposits pheromone on the anterolateral surfaces of vertical structures in her environment. The male sensorily assays the sides of these objects and from this information determines the female's direction of travel.     

Sex pheromone ofPlanotortrix species found on mangrove

(Z)-5-Tetradecenyl acetate and tetradecyl acetate were identified as sex pheromone components of an unnamedPlanotortrix leafroller moth species found onAvicennia resinifera (mangrove). An equal mixture of the two compounds used as bait gave field trap catches at least as good as those baited with caged virgin females. Traps baited with the two chemicals caught malePlanotortrix moths in a mangrove swamp not previously found to host the unnamedPlanotortrix species. Adults of the unnamedPlanotortrix species and of the greenheaded leafroller,Planotortrix excessana are morphologically indistinguishable. The sex pheromone ofP. excessana has been found previously to be a mixture of (Z)-S-tetradecenyl acetate and tetradecyl acetate, and this means that the two species may now be distinguished by sex pheromone differences.Lepidoptera: Tortricidae: Tortricinae.     

Sex pheromone of oriental armywormMythimna separata Walker

In the mainland of China, the male Oriental armyworm was not attracted to the sex pheromone components (Z)-11-hexadecenyl acetate and (Z)-11-hexadecenol identified by Takahashi et al. in 1979. By means of EAG, GC, and GC-MS techniques, (Z)-11-hexadecenal, hexadecenal, and (Z)-11-hexadecenol were found in female gland washings, and encouraging captures were obtained in preliminary field trapping.     

Sex pheromone of European corn borer.

Sex gland extracts ofOstrinia nubilalis females collected in the wild or laboratory-reared from Switzerland, Italy, and Hungary were analyzed. Individuals collected in the north of Switzerland contained (Z)- and (E)-11-tetradecenyl acetate at the approximate ratio of 973 (Z type), in accordance with field responses of males and previous findings. On the other hand, females from a laboratory culture derived from field collections made in the same area and reared for four to five generations contained theZ andE isomers at ratios of ca. 397 and 3565, respectively. In the south, one of the eight wild females analyzed was of theZ type and the rest intermediate, whereas males were predominantly trapped with blends of the two isomers containing 60 to 97E. In a laboratory culture reared for one to two generations from corn borers collected in Hungary, three of nine females were of the intermediate type and the restZ. Small amounts of (Z)-11-hexadecenyl acetate were detected in female glands of theE strain; however, no effect of this compound could be observed in the field.     

Selection of a polyethylene tubing formulation of (Z)-9-tetradecen-1-ol formate and its use in disrupting pheromone communication inHeliothis zea (Boddie)

The release rates of (Z)-9-tetradecen-1-ol formate (Z9TDF) from four sizes of polyethylene tubing were measured periodically for 50 days by collecting the Z9TDF vapors on a polymeric adsorbent. The resulting release rate curves had shapes that were determined by the tubing's diameter and wall thickness. One tubing size that combined good longevity with adequately high release rates was tested in the field to determine whether it could disrupt pheromone communication of the corn earworm,Heliothis zea (Boddie). We observed a low percentage of disruption, but release rate measurements made concurrently showed that the tubing was releasing Z9TDF normally. In a second field test, a larger quantity of Z9TDF was released, and 87% disruption was attained for 66 days.Mention of a commercial or proprietary product in this paper does not constitute endorsement of that product by the USDA.     

Effect of release rate and ratio of (Z)-11-hexadecen-1-ol from synthetic pheromone blends on trap capture ofHeliothis subflexa (Lepidoptera: Noctuidae)

Response of maleHeliothis subflexa to pheromone-baited traps containing blends of tetradecanal, (Z)-9-tetradecanal, hexadecanal, (Z)-7-hexadecenal, (Z)-9-hexadecenal, (Z)-11-hexadecenal, hexadecan-1-ol acetate, (Z)-7-hexadecen-1-ol acetate, (Z)-9-hexadecen-1-ol acetate, (Z)-11-hexadecen-1-ol acetate, (Z)-9-hexadecen-1-ol, and (Z)-11-hexadecen-1-ol was evaluated. Analysis of trap capture data indicated that (Z)-11-hexadecen-1-ol was a critical component of the pheromone blend. It was determined from emission rate data and measurements of the ratio of pheromone components emitted from rubber septa tested that a significant increase in trap capture of maleH. subflexa occurred when the blends investigated released the alcohol in a narrow range relative to the total amount of pheromone emitted. The optimum range of release ratio of the alcohol for the capture of males in sticky traps was determined to be 0.9–3.5% of the pheromone blend. This release ratio range was reduced to 0.9–1.6% when bucket traps were used.     

INTROGRESSING PHEROMONE QTL BETWEEN SPECIES: TOWARDS AN EVOLUTIONARY UNDERSTANDING OF DIFFERENTIATION IN SEXUAL COMMUNICATION

As a first step toward understanding how noctuid moths evolve species-specific pheromone communication systems, we hybridized and backcrossed two closely related moth species, Heliothis virescens (Hv) and H. subflexa (Hs), which differ qualitatively and quantitatively in their multi-component sex pheromone blends. We used amplified fragment length polymorphism (AFLP) marker-based mapping of backcross families to determine which of the 30 autosomes in these moths contained quantitative trait loci (QTL) controlling the percentages of specific chemical components in the pheromone blends. In two previous backcrosses to Hs, we found a strong depressive effect of Hv-chromosome 22 on the percentage of three acetate components in the pheromone gland. These acetates are present in Hs and absent in Hv. Here, we describe how we introgressed Hv-chromosome 22 into the genomic background of Hs. Selection for Hv-chromosome 22 started from backcross 3 (BC₃) females. All females that had Hv-chromosome 22 and a low percentage of acetates (< 3% of the total amount of pheromone components present) were backcrossed to Hs males. In BC₅ to BC₈, we determined whether Hv-chromosome 22 was present by a) running only the primer pairs that would yield the markers for that chromosome, and/or b) determining the relative percentages of acetates in the pheromone glands. Either or both genotype and phenotype were used as a criterion to continue to backcross these females to Hs males. In BC₉, we confirmed the isolation of Hv-chromosome 22 in the Hs genomic background, and backcrossed the males to Hs females to eliminate the Hv-sex chromosome as well as mitochondrial DNA. The pheromone composition was determined in BC₃, BC₅, and BC₁₁ females with and without Hv-chromosome 22. All backcross females with Hv-chromosome 22 contained significantly less acetates than females without this chromosome. In addition, BC₃ females with Hv-chromosome 22 contained significantly more Z11-16:OH than BC₃ females without Hv-chromosome 22. However, in BC₅ and BC₁₁ females, the correlation between Z11-16:OH and Hv-chromosome 22 was lost, suggesting that there are separate QTL for the acetates and for Z11-16:OH, and that the relative amount of the alcohol component is only affected in epistasis with other (minor) QTL. Now that we have succeeded in isolating the chromosome that has a major effect on acetate production, we can test in behavioral experiments whether the presence of acetates may have been a driving force for a shift in pheromone composition. Such tests are necessary to move towards an evolutionary understanding of the differentiation in sexual communication in Heliothis spp. moths.     

Sex pheromone chemistry of female corn earworm moth,Heliothis zea

Glass open-tubular capillary Chromatographic and combined glass open-tubular capillary chromatographic-mass spectrometric analyses of ovipositor washes of femaleHeliothis zea, chemical characterization, chemical synthesis, and laboratory and field bioassays showed that the ovipositor wash of the species is made up of 90–95% (Z)-11-hexadecenal, 1–2% (Z)-9-hexadecenal, 0.4–2% (Z)-7-hexadecenal, and 2–7% hexadecanal. Stimuli containing a binary mixture of (Z)-11-hexadecenal and (Z)-9-hexadecenal or the binary mixture in combination with any of the other aldehydes identified from the females elicited intense attraction and close-range precopulatory reactions fromH. zea males.Mention of a commercial or proprietary product in this paper does not constitute a recommendation or an endorsement of that product by the U.S. Department of Agriculture.     

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.     

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.     

Sex pheromone of fall armyworm

Two components of the fall armyworm,Spodoptera frugiperda (J.E. Smith), sex pheromone, (Z)-9-dodecen-1-ol acetate (DDA) and (Z)-9-tetradecen-1-ol (TDA), were tested alone and in combination to determine their effects on male sexual response and inhibition of mating in the laboratory. The threshold response for FAW males was lower for TDA than for DDA, and males responded to TDA over a wider range of dosages. Although TDA is not attractive to FAW males in the field, this compound was highly effective in reducing mating under laboratory conditions.cooperating with the Insect Attractants, Behavior, and Basic Biology Research Laboratory, Gainesville, Florida 32604.     

Effect of formulations and dispensers on attractiveness of virelure to the tobacco budworm

Formulations of virelure, the synthetic sex pheromone of the tobacco budworm,Heliothis virescens (F.), were developed, bioassayed in traps in field cages, and used in traps to survey populations in plant hosts. Virelure (10 mg) laminated between thin sheets of vinyl polymer plastic was attractive to male moths for 21 days, and attractiveness did not regress appreciably for the first 7 days. Another dispenser was an 8 × 30-mm cigarette filter encased in a glass shell vial. A mixture of 10 l (Z)-11-hexadecenal and 0.5 l (Z)-9-tetradecenal, the pheromone components, formulated with wheat germ oil andd-alpha tocopherol (91) in 0.5 ml CH₂Cl₂ (WGOE) was still attractive to males after 5 days. Cottonseed oil, polyethylene glycol 600 distearate (peg-600-d), and peg-6000-d also inhibited excessive vaporization and oxidation of virelure. Virelure (21.25 l) formulated on filters with WGOE and used as bait (replaced weekly) in survey traps caught 27,263 males in 1975; analysis of catches each day for 7-day periods showed that most males were caught within 3 days after baiting, and attractiveness regressed considerably during the last 3–4 days of each week. Four ratios of virelure components were tested as bait (20 mg) in saucer-type pheromone traps, and the 161 ratio caught significantly more male moths than ratios of 241, 321, or 401.Lepidoptera Noctuidae.In cooperation with the Texas Agricultural Experiment Station, Texas A & M University, College Station 77843.This paper reports the results of research only. Mention of a pesticide in this paper does not constitute a recommendation for use by the USDA nor does it imply registration under FIFRA as amended. Also, mention of a commercial or a proprietary product in this paper does not constitute an endorsement by the USDA.     

Evidence for the controlled release of a crustacean sex pheromone

Evidence for the controlled release of a crustacean sex pheromone: mature malePortunus sanguinolentus (Herbst) did not give the characteristic display described by Ryan (1966) to water from a container with a carried or artificially restrained premolt female. However, the sex pheromone was still present in the urine of the restrained female, suggesting that she inhibits the emission of the sex pheromone by regulating her urine flow.Based on part of a dissertation presented by the author in partial fulfillment of the requirements for the doctoral degree in zoology at the University of Hawaii.