Pheromone Titer Research Articles

Effect of tyramine and stress on sex-pheromone production in the pre- and post-mating silkworm moth, Bombyx mori

Tyramine (TA) increased significantly after mating, whereas there were no significant differences in octopamine (OA) and dopamine (DA) levels in the brain–suboesophageal ganglion (SOG) complexes between virgin and mated females. The effects of various biogenic amines were tested on pheromone production of virgin and mated females of the silkworm moth, Bombyx mori. After 8 h a significant reduction by TA (46%) was observed. Meanwhile, when OA or DA was injected, a significant increase of pheromone titer was observed in both virgin and mated females. This study also presents evidence for an increase in levels of OA and DA in the brain–SOG complexes in response to mechanical stress in B. mori female. TA suppressed pheromone production in an in vitro pheromone gland (PG) homogenate preparation, thus suggesting that the target of TA is the PG. TA inhibited pheromone production in vitro in a dose-dependent manner and DA had a lower inhibitory activity than TA, whereas OA had no effect, suggesting that TA is a candidate for regulating pheromone production in the PG, although other factors could be responsible for the pheromonostatic function.

Mechanisms of premating isolation between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

Helicoverpa armigera and Helicoverpa assulta are sympatric sibling species, and in the laboratory they can interbreed and produce viable offspring. To assess the contributions of temporal barriers and sexual barriers to premating isolation, we investigated both the temporal rhythms of calling behavior and pheromone titers of H. armigera and H. assulta females and the behavioral responses of males to conspecific and heterospecific calling females in a wind tunnel. Both H. armigera and H. assulta females called throughout the scotophase, and there was more calling during the second half of the scotophase than during the first half. Maximal pheromone titer and maximal calling activity in H. armigera synchronously occurred at the sixth hour into the scotophase, whereas, in H. assulta, the maximal pheromone titer occurred 2 h before the peak of calling. Pheromone blend ratios of the two species were opposite and, within each species, changes in the ratio within the scotophase and at different ages were relatively small. Males of both H. armigera and H. assulta responded strongly to their conspecific calling females in the wind tunnel and completed the whole courtship sequence. In contrast, they did not land and had no copulation attempts in response to heterospecific calling females. These results show that the two species do not have obvious temporal differences in calling behavior and pheromone production, and the specificity of sex pheromone blend emitted by females plays a key role in their premating isolation. In addition, we summarized the potential isolation mechanisms of H. armigera and H. assulta.

Heritable Variation in the Sex Pheromone of the Almond Moth, Cadra cautella

The sex pheromone of the female almond moth, Cadra cautella, constitutes a highly variable signal. Age and period of the L/D cycle had significant effects on the variation in titers of (Z,E)-9,12-tetradecadienyl acetate (Z9,E12-14:Ac) and (Z)-9-tetradecenyl acetate (Z9-14:Ac) and the ratio of Z9,E12-14:Ac to Z9-14:Ac. Across age classes, minimum and maximum mean titers (ng +/- SE) of Z9,E12-14:Ac and Z9-14:Ac ranged from 8.7 +/- 0.46 to 21 +/- 1.0 and from 2.0 +/- 0.14 to 2.9 +/- 0.15, respectively; the mean ratio of Z9,E12-14:Ac to Z9-14:Ac ranged from 5.2 +/- 0.21 to 11 +/- 0.40. The titers of Z9,E12-14:Ac and Z9-14:Ac and the ratio of Z9,E12-14:Ac to Z9-14:Ac were highest at the onset of the scotophase and lowest at the onset of the photophase. Similarly, the titer of Z9,E12-14:Ac and the ratio of Z9,E12-14:Ac to Z9-14:Ac were highest in females sampled 1 d postemergence and declined over all age classes. The titer of Z9-14:Ac increased from day 1 to day 2, and then declined to levels equivalent to day 1. Analysis of pupal and adult mass demonstrated a positive correlation (r = 0.874, P < 0.001); however, the titer of neither Z9,E12-14:Ac or Z9-14:Ac nor the ratio of Z9,E12-14:Ac to Z9-14:Ac were significantly correlated with pupal mass. Age-related variations in pheromone titer and ratios were heritable. The narrow-sense heritability of the observed variation in 1-d-old females, 1 hr into the scotophase is 1.2 +/- 0.32, 0.75 +/- 0.24, and 0.46 +/- 0.17 for the titer of Z9,E12-14:Ac and Z9-14:Ac and the ratio of Z9,E12-14:Ac to Z9-14:Ac, respectively. A significant additive genetic correlation was observed between that of Z9,E12-14:Ac and Z9-14:Ac, but not between that of either Z9,E12-14:Ac or Z9-14:Ac and the ratio of Z9,E12-14:Ac to Z9-14:Ac. The results of the genetic analyses suggest that the traits of titer of Z9,E12-14:Ac and Z9-14:Ac have a greater potential to respond to selection than the trait of ratio of Z9,E12-14:Ac to Z9-14:Ac.

Circadian Rhythms of Sexual Behavior and Pheromone Titers of Two Closely Related Moth Species Autographa gamma and Cornutiplusia circumflexa1

Two closely related plusiinae moths, Autographa gamma Linnaeus and Cornutiplusia circumflexa Linnaeus, are sympatric in Israel. Both species use identical sex pheromone components but in different ratios, and do not attempt to mate with each other. In addition to the effective reproductive separation by their sex pheromones, the sexual behavior of both species was compared to determine whether the lack of selection pressure might create additional barriers to cross-attraction and cross-mating. We found the gamma moth to be sexually active almost equally throughout the scotophase, whereas the sexual activity of C. circumflexa was limited to a short period at the end of the scotophase when most of the gamma moths had already mated. Higher levels of calling were observed with older females. There was a close relationship between pheromone titer and calling activity in both species.

The fate of topically applied fatty acids in the sex pheromone gland of the moth Heliothis virescens

Deuterium-labeled hexadecanoic acid (D4–16:COOH), a sex pheromone biosynthetic intermediate, and heptadecanoic acid (D3–17:COOH), an acid that cannot be converted to sex pheromone, were topically applied to the pheromone gland of female Heliothis virescens, and the fate of the label determined. Both acids were incorporated similarly into the glycerolipids, with by far the greatest amount found in the triacylglycerols (TGs), and relatively small amounts found in other neutral and polar classes. For D4–16:COOH, the labeled pheromone precursor, ( Z)-11-hexadecenoate, was also found predominantly in the TGs but relatively (compared to labeled hexadecanoate) high amounts were also found in the phospholipids. Within the TGs, both acids, as well as the pheromone precursor, were found almost exclusively on the sn-3 position of the glycerol backbone. This demonstrates that the major fate, in the glycerolipids, of free fatty acids is addition to 1,2-diacylglycerols. A relatively large amount of the applied acid was also found in the gland in the form of the acyl-CoA thioester. In a 24-h time–course study, this form remained at a relatively high level for the duration of the assay, and decreased at a rate comparable to the titer of this acid in the TGs, suggesting that titers of fatty acids in the glycerolipids and acyl-CoA thioesters may be in equilibrium. A time–course assay with D4–16:COOH demonstrated that peak pheromone titer after application was reached before peak titers of both total hexadecanoate and hexadecanoyl-CoA. Combined with a dose–response experiment, which showed that labeled pheromone titer did not increase above an applied concentration of 20 mg/ml, these data suggest that the final step in pheromone biosynthesis, reduction of Z11–16:Acyl-CoA, may be inhibited by increased acyl-CoA titers in the gland. Overall, our data are consistent with the glycerolipids modulating acyl-CoA concentrations in the pheromone gland.

Lipid analysis of the sex pheromone gland of the mothHeliothis virescens

The sex pheromone gland of female Heliothis virescens was analyzed for fatty acid and lipid content. Base methanolysis of the gland showed a large amount of methyl (Z)-11-hexadecenoate (Z11-16:Acyl), the fatty acyl analog of the major pheromone component, (Z)-11-hexadecenal, as well as a small amount of methyl (Z)-11-octadecenoate. Methyl esters of various common fatty acids were also observed. HPTLC analysis of the glandular lipids revealed large quantities of triacylglycerols (TGs), and lesser amounts of 1,2-diacylglycerols (1,2-DGs), 2-monoacylglycerols (2-MGs), phosphatidyl ethanolamines, and phosphatidyl cholines. The greatest amount of Z11-16:Acyl in these lipids was in the TGs, with lesser amounts in the two phospholipid classes and only trace amounts in the other neutral lipids. The glands of females at various ages and photoperiodic times were extracted, fractionated into neutral and polar fractions by silica SPE, and fatty acid titers in these fractions determined. All fatty acids, but notably Z11-16:Acyl, showed significant total and neutral lipid fraction peaks at mid scotophase for 2-day-old females; a less dramatic, but significant, Z11-16:Acyl peak in the polar fraction was also observed. However, only a relatively small proportion (<50%) of this acid was recovered from the silica at all times. This "non-recoverable" Z11-16:Acyl showed a dramatic and significant peak at mid scotophase for 2-day females, corresponding roughly with maximal pheromone titer. All other acids in the gland were recovered in high proportions, and their respective "non-recoverable" titers were not different at any of the times analyzed. Based on previous work, this non-recoverable Z11-16:Acyl is likely the CoA ester. Therefore, it appears that the pheromone gland of H. virescens maintains pools of Z11-16:Acyl in both CoA ester and TG forms, which are available for biosynthesis of pheromone. These pools are greatest during maximal pheromone production when the biosynthetic enzymes, possibly the fatty acid reductase, are unable to utilize rapidly enough the quantities of Z11-16:Acyl biosynthesized.

Sublethal effects of larval treatment with deltamethrin on moth sex pheromone communication system of the Asian corn borer, Ostrinia furnacalis

The first, third, and fifth instar larvae of Asian corn borer, Ostrinia furnacalis (Guenée) (Lepidoptera: Pyralidae), were treated with a lethal dose 30% of deltamethrin to determine its sublethal effects on chemical communication of the adult survivors. The capability of all males surviving deltamethrin to contact the pheromone source in a wind tunnel was significantly reduced, but those treated as the first and third instars also exhibited lower percentages of `orientation,' `up-wind flight,' and `approaching' behaviors than the control males. Female moths from the deltamethrin treatment exhibited similar calling periodicity and percentages to the control moths during 10-h scotophase. However, approximately 20% of the females that survived the pyrethroid in the first and third instars contained significantly higher amounts of sex pheromone and wider coefficients of variation of the ratio of E12-tetradecenyl acetate to its Z isomer in those female survivors. There was no significant difference in the pheromone titers and coefficients of variation in control females and the females treated in the fifth instars. The present study indicates that the Asian corn borer may possess a compensation system by which the males that survived treatment at larval stage present a lower response to the pheromone, while the female survivors produce and release more pheromone.

Diel periodicity and influence of age and mating on female sex pheromone titre in Estigmene acrea (Lep., Arctiidae)

Abstract: Calling behaviour, diel periodicity, and effect of age and mating on female sex pheromone titre in Estigmene acrea (Drury) were studied under laboratory conditions. Forty‐five per cent of females started calling during the first scotophase, but the highest number of calling females was observed during the second, third and fourth scotophases. Calling behaviour occurred from the third hour after dark until just before the end of the scotophase. However, females exhibited a bimodal pattern of calling with the first peak occurring between 4 and 6 h and a second peak at 10 h after the onset of scotophase. The mean onset of calling time differed significantly with age. Older females showed a tendency to call longer, but there was no significant difference. The amount of (Z,Z)‐3,6‐cis‐9, 10‐epoxyheneicosadiene in females was quantified from the first scotophase following emergence, until the fifth scotophase. Glands of 0‐day‐old females presented a higher content of pheromone compared with that found in glands of 1‐, 2‐, 3‐ and 4‐day‐old females. Pheromone titre was determined at 2‐h intervals throughout the third scotophase and photophase. (Z,Z)‐3,6‐cis‐9,10‐epoxyheneicosadiene was found in the gland during the scotophase as well as the photophase. However, there was no consistent pattern of pheromone production throughout the scotophase or photophase. Mated females of E. acrea produced significantly less pheromone than virgin females.

Fatty acid and sex pheromone changes and the role of glandular lipids in the Z-strain of the European corn borer, Ostrinia nubilalis (Hübner).

Lipids in the sex pheromone gland of females of the Z-strain of Ostrinia nubilalis were analyzed for fatty acyl pheromone analogs (FAPAs) and other potential biosynthetic intermediates. More than 80% of the FAPAs were found in the triacylglycerols (TGs), with smaller amounts found in the phosphatidyl cholines, ethanolamines, and serines. Analysis of the TGs by lipase revealed that the two FAPAs were distributed fairly evenly among all three stereospecific positions. Comparison of changes in titers of key glandular fatty acids with those of pheromone components, with respect to photoperiodic time and age of females, showed that both FAPA and pheromone titers exhibited a cyclical pattern with peaks in the scotophase and valleys in the photophase. However, whereas pheromone titer tended to peak in the first half of the scotophase, FAPA titer peaked at the end of the scotophase. Significantly, the titer of the FAPA of the minor component, (E)-11-tetradecenyl acetate (3% of pheromone), was always much greater than the titer of the FAPA of the major component, (Z)-11-tetradecenyl acetate (97%), of the pheromone. Titer of myristate, an intermediate in pheromone biosynthesis, was also higher during the scotophase than the photophase. However, myristate titer showed a pronounced dip in the middle of the scotophase. These data suggest two roles for glandular lipids in sex pheromone biosynthesis in O. nubilalis. Firstly, they remove excess FAPA of the minor component so the fatty acid reductase system is not presented with a high ratio of this isomer (which would otherwise result from the reductase's own selectivity), which could cause changes in the final pheromone ratio. Secondly, hydrolysis of the large amounts of stored saturated fatty acids from the TGs may provide substrate for pheromone biosynthesis.

Pheromone analysis of wild female moths with a PBAN C-terminal peptide injection for an estimation of assortative mating in adzuki bean borer, Ostrinia scapulalis.

The adzuki bean borer, Ostrinia scapulalis, has distinct genetic variation in the blend of two sex pheromone components, (E)- and (Z)-11-tetradecenyl acetates. This variation is largely controlled by a single autosomal locus with two alleles, E and Z. E-type (EE) females produce a pheromone with a mean E:Z ratio at 99:1 whereas Z-type (ZZ) and I-type (ZE) produce pheromones with mean of 3:97 and 64:36, respectively. Interestingly, in many natural populations of O. scapulalis in Japan, this pheromone polymorphism appears to be stably maintained. We tried to predict the changes in relative abundance of each pheromone type by estimating the pheromone production genotype of wild females and their male mates. The pheromone titer in the wild, mated females was increased without changing the blend ratio by an injection of a peptide with pheromone biosynthesis activating activity (TKYFSPRL-NH2). The frequencies of E-, I-, and Z-types at Matsudo were 15, 52, and 33%, respectively, and did not deviate from the Hardy-Weinberg expectations. The estimated mating patterns were concordant with the assumption that no assortative mating was occurring in this population, and this is suggested as a cause of sustained polymorphism at Matsudo.

Control of the Oriental Fruit Moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae) by Mating Disruption with Sex Pheromone in Pear Orchards

The effect of dispensers containing a 93: 6 : 1 blend of Z8-12: Ac, E8-12: Ac, and Z8–12: OH for mating disruption of Grapholita molesta (Busck) was tested from 1998 to 2000 in pear orchards in Naju, Korea. The permeation of the atmosphere with synthetic sex pheromone significantly reduced the ability of male moths to locate pheromone-baited traps, and reduction of male moths to locate pheromone-baited traps was not significantly different among different pheromone doses. Success in suppression of infestations by pheromone permeation was achieved under the dose rate of 50 g/ha, and increasing pheromone concentration above this dose did not significantly reduce the fruit damage. The efficacy of pheromone treatment could not be predicted using captures of males in traps baited with synthetic pheromone. Apparently, a much lower titer of pheromone was required to reduce trap catch than to reduce damage. The trap catch was influenced by trap position in pheromone plots; significantly more moths were caught at the outer areas than at the inner areas. Also, fruit damage levels in pheromone plots were significantly heavier in the trees located in the outer area. However, the lowered efficacy in the outer areas was partly improved by the application of a high number of dispensers at these areas compared with the inner areas. Assessment of trap catch and damage level over a three-year period showed that the effectiveness of the disruption technique for G. molesta was enhanced through sequential suppression year after year at the same site.

Effects of sublethal deltamethrin on the chemical communication system and PBAN activity of Asian corn borer, Ostrinia furnacalis (Güenee).

Topically applied sublethal doses of deltamethrin may interrupt chemical communication between the two sexes of the Asian corn borer, Ostrinia furnacalis (Güenee), by affecting the probability of calling behavior, sex pheromone titer and PBAN-like activity. At the 1st, 2nd, and 4th day of post-treatment with deltamethrin (1 ng/moth), female O. furnacalis showed 32, 36, and 46% calling probabilities of control female moths, while the sex pheromone titer of females was only 15, 20, and 25% the amount of control females, respectively. The chemical communication system was significantly affected even at the 4th day post-treatment. By injections of 1, 1/2, 1/4, and 1/8 female equivalent of brain and subesophageol ganglion homogenate of 1 ng deltamethrin-treated females into decapitated females, female pheromone production was reduced to 64, 31, 30.7, and 8.1% of that of control female moths, respectively. This result appears to indicate that sublethal deltamethrin affected the activity of the PBAN-like factor of O. furnacalis.

Age-related changes in the competency of the pheromone gland and the pheromonotropic activity of the brain of both virgin and mated females of two Choristoneura species

Nine-day-old decapitated females injected with different doses of Hez-PBAN produced significantly less pheromone than 1-day-old individuals, suggesting that the age-related decline in the pheromone titre of Choristoneura fumiferana and C. rosaceana virgin females was primarily the result of a reduced ability of the glands to produce pheromone. In C. fumiferana, lower pheromonotropic activity of the Br-SEG may also contribute to the pheromone decline with age but not in C. rosaceana, as the pheromonotropic activity of the Br-SEG remained constant throughout the females’ life. In both Choristoneura species, mating also suppressed pheromone production (pheromonostasis) after 24 h. The Br-SEG of mated females contained PBAN but there was no indication that its concentration changed with time post-mating since Br-SEG homogenates obtained from different-aged mated females showed the same level of pheromonotropic activity in both Choristoneura species. However, as observed in virgins, pheromone glands of older mated females were less sensitive to PBAN than those of younger ones. This suggests that the probability of Choristoneura females to attract a second mate may decrease with an increase in the refractory period following the first mating.

Mating Effect on Sex Pheromone Production of the Oriental tobacco budworm, Helicoverpa assulta

This study was undertaken to clarify the suppression phenomenon of sex pheromone production after mating and its relationship to the physiological mechanism in adult females of Helicoverpa assulta, and determine the mating factor from males causing depletion of sex pheromonc production. Sex pheromone production of H. assulta females was mostly terminated in 3 hours after mating. Mated females maintained with a low titer of sex pheromone until 3 days when it started to increase again, which showed a characteristic of species mating more than once. The mated female again produced pheromone upon injection of pheromone biosynthesis activating neuropeptide (PBAN) or extracts of brain-suboesophageal ganglion complexes (Br-Sg) of mated female, which were shown similar pheromonotropic activities as compared with virgin females. These results indicated that the mating did not inhibit the receptivity of pheromone gland itself and PBAN biosynthesis in suboesophageal ganglion of the mated females. And it seems to support that the depletion of sex pheromone production is responsible for blocking of PBAN release from head. To investigate the mating factor from adult males, when extracts of reproductive organs of male were injected into hemocoel of virgin females evoking depletion of sex pheromone production as shown in mated female. The results suggest that a chemical substance(s) from the male reproductive organs could be responsible for the loss of sex pheromone biosynthesis in H. assulta.

Stimulation of sex pheromone production by PBAN‐like substance in the pine caterpillar moth, Dendrolimus punctatus (Lepidoptera: Lasiocampidae)

Sex pheromone production in the female pine caterpillar moth, Dendrolimus punctatus is controlled by a PBAN-like substance located in the head of female moth. Pheromone titer was significantly decreased by decapitation of female moth, and restored by injection of either Hez-PBAN or head extract prepared from male or female moth. Stimulation of pheromone production by head extract followed a dose-dependent pattern from 0.5 to at least 4 head equivalent. A gland in vitro assay was used to study the relationship between gland incubation time and pheromone production as well as calcium involvement in the stimulation of pheromone production by head extract. Maximum pheromone production was occurred at 60 min after pheromone gland was incubated with two equivalents of head extracts. In vitro experiments showed that the presence of calcium in the incubation medium was necessary for stimulation of pheromone production. The calcium ionophore, A 23187, alone stimulated pheromone production. The pheromone components (Z,E)-5,7-dodecadienol and its acetate and propionate were produced in these experiments but in addition to the aldehyde, (Z,E)-5,7-dodecadienal was also found. This indicates that females are capable of producing four oxygenated functional groups. The PBAN-like substance control of the pheromone biosynthetic pathway was investigated by monitoring the incorporation of the labeled precursor into both pheromone and pheromone intermediates.Arch. Insect Biochem. Physiol. 49:137–148, 2002. © 2002 Wiley-Liss, Inc.

Stimulation of sex pheromone production by PBAN-like substance in the pine caterpillar moth, Dendrolimus punctatus (Lepidoptera: Lasiocampidae).

Sex pheromone production in the female pine caterpillar moth, Dendrolimus punctatus is controlled by a PBAN-like substance located in the head of female moth. Pheromone titer was significantly decreased by decapitation of female moth, and restored by injection of either Hez-PBAN or head extract prepared from male or female moth. Stimulation of pheromone production by head extract followed a dose-dependent pattern from 0.5 to at least 4 head equivalent. A gland in vitro assay was used to study the relationship between gland incubation time and pheromone production as well as calcium involvement in the stimulation of pheromone production by head extract. Maximum pheromone production was occurred at 60 min after pheromone gland was incubated with two equivalents of head extracts. In vitro experiments showed that the presence of calcium in the incubation medium was necessary for stimulation of pheromone production. The calcium ionophore, A 23187, alone stimulated pheromone production. The pheromone components (Z,E)-5,7-dodecadienol and its acetate and propionate were produced in these experiments but in addition to the aldehyde, (Z,E)-5,7-dodecadienal was also found. This indicates that females are capable of producing four oxygenated functional groups. The PBAN-like substance control of the pheromone biosynthetic pathway was investigated by monitoring the incorporation of the labeled precursor into both pheromone and pheromone intermediates.

Diel rhythms of calling behavior and temporal change in pheromone production of the rice leaffolder moth, Cnaphalocrocis medinalis(Lepidoptera: Crambidae).

Calling activity in Cnaphalocrocis medinalis showed a distinct diel periodicity. Moths showed high activity during 5–7 h into the scotophase. To clarify the temporal change of pheromone titers in the pheromone gland within 1 d, age 4 females were used. The changes in the calling activity and titer of the four sex pheromone components, (Z)-11-octadecenal (Z11-18:Ald), (Z)-13-octadecenal (Z13-18:Ald), (Z)-11-octadecen-1-ol (Z11-18:OH) and (Z)-13-octadecen-1-ol (Z13-18:OH), in the pheromone glands were roughly synchronous. At age 4, however, the pheromone components could already be detected at the onset of scotophase, while calling began at 3 h into the scotophase. Furthermore, calling was most active between age 3 and age 7, whereas the highest level of pheromone titer at 6 h after light off was observed from age 3 to age 5. These findings suggest the presence of different controlling mechanisms between calling behavior and pheromone production. The ratio of the four sex pheromone components remained relatively constant at all times of the day and all ages.

Factors involved in the post-copulatory neural inhibition of pheromone production in Choristoneura fumiferana and C. rosaceana females

Normal mating lasts approximately 3 h in Choristoneura fumiferana and C. rosaceana. Data generated from interrupted matings showed that the act of mating did not suppress pheromone production (pheromonostasis) in either species although, in C. rosaceana, pheromone titre declined slightly the night following mating. In both species the migration of sperm to the spermatheca (SP) occurred several hours after mating, and coincided with a significant and permanent depression in pheromone titre, as well as egg fertilisation and oviposition. However, disrupting matings within 2 h of the onset resulted in oviposition patterns similar to virgins in both species, with mostly infertile eggs being laid by C. fumiferana females while oviposition was totally inhibited in C. rosaceana. The transection of the ventral nerve cord (VNC) 1 h post-mating did not result in the depression of pheromone titres the following night in either species but if the VNC was transected 3 h post-mating, pheromonostasis was observed. While 25% of C. fumiferana females had sperm in their SP 2 h after mating, it took at least 4 h in C. rosaceana. This suggests that while the physical presence of sperm in the SP may play some role in the termination of pheromone production in C. fumiferana, other factors must trigger the neural signal that elicits pheromonostasis in both species. A better understanding of the temporal dynamics of both apyrene and eupyrene sperm within the different parts of the female reproductive system might clarify these interspecific differences.

Sex pheromone levels in pheromone glands and identification of the pheromone and hydrocarbons in the hemolymph of the moth Scoliopteryx libatrix L. (Lepidoptera: Noctuidae).

The hydrocarbon sex pheromone (13-methyl-Z6-heneicosene) of Scoliopteryx libatrix L. (Lepidoptera: Noctuidae) was found to reach its highest levels on pheromone glands of 3-day-old females. Pheromone levels were not different between the time of maximum calling (end of scotophase) and at the middle of photophase. Overwintering females collected in October had sex pheromone present. Decapitation did not lower the amount of pheromone present, indicating that a head factor is not involved in maintaining pheromone titers. Hemolymph also contained the pheromone, indicating that it is made by oenocytes and transported to the sex pheromone gland. Longer chain length hydrocarbons were also identified from the hemolymph and on the cuticular surface. Quantitative differences in hydrocarbon profiles were found with more methyl-branched hydrocarbons found in the hemolymph than on the cuticular surface. Arch.

DIEL RHYTHMS OF CALLING BEHAVIOR AND SEX PHEROMONE PRODUCTION OF BEET ARMYWORM, SPODOPTERA EXIGUA (LEPIDOPTERA:NOCTUIDAE)*

Abstract Beet armyworm (BAW), Spodoptera exigua, is becoming one of more and more serious pests in China in recent years. As a part of research program of sex pheromone and its application of BAW in China, the hourly and daily variation of calling behavior and pheromone production of BAW females were investigated. Both calling behavior and titers of 4 sex pheromone components showed distinct diel rhythms, and the two peak periods were synchronous. In comparison, the calling activity lasted shorter period of time with a longer peak time, whereas the production of the sex pheromone lasted throughout the whole scotophase and part of the photophase with a very short peak time. The calling behavior began at the middle scotophase, reached the maximum at the middle‐later scotophase, and continued the maximal calling activity until the end of the scotophase. When the light was on, the calling percentage reduced sharply, and all females stopped calling 1 hour later. The variation patterns of the 4 pheromone components in the glands of the 3 day old moths were similar from one to another. From 0.5 h before to 4.5 h into scotophase, the titers increased slightly, but at 6.5 h they showed a significant increase up to the peak values of the scotophase. Soon at 8.5 h into the scotophase, they decreased significantly and thereafter gradually to undetectable level at 4.5 h into the next photophase. The daily change experiment showed that BAW females began calling at 0 scotophase, became fully active in calling at 3rd scotophase, and maintained the calling activity to 7th scotophase. There was no significant difference in pheromone titers among different day‐old moths.