Ecdysone and a dietary alkaloid interact in the development of the pheromone gland of a male moth (Creatonotos,Lepidoptera: Arctiidae)

Summary Hair-covered scent organs of the male arctiid moth Creatonotos produce and dissipate the volatile pheromone hydroxydanaidal. The biosynthesis of this substance depends quantitatively upon the uptake of pyrrolizidine alkaloids (PA) with the larval foodplant. The size of the tubular, eversible scent organ (corema) is also positively correlated with the ingested amount of the same alkaloid, which acts like a specific growth factor. After an assessment of the corema normogenesis by Rick-Wagner (PhD thesis, University of Cologne, 1986) we injected PA into PA-free raised larvae, prepupae, and pupae. We found that the PA competence (sensitivity) of the corema anläge terminates with the first prepupal day. Ecdysone titer determinations (radioimmunoassay) are in agreement with those in other moth species. Ligated (ecdysone-free) pupal abdomina never developed imaginal structures, with or without earlier PA application. Ecdysone injection into ligated pupal abdomina of PA-fed specimens initiated the development of imaginal structures and also of coremata of more than (ecdysone-free) control size. Pupal abdomina without PA pre-treatment only developed very small coremata. With these experiments we have separated and identified two morphogenetic control agents of corema development: the dietary PA specifies the size of the organ whereas ecdysone induces the anlage to proliferate within these PA-dependent ranges and to differentiate adult structures, as it does with other imaginal anlagen.     

Biotransformation of linoleic acid with the Candida tropicalis M25 mutant

Linoleic acid was transformed by mutant Candida tropicalis M25 and transformations were studied in batch and fed-batch cultures. Cofermentations with palmitic acid as inducer of the fatty acid degradation pathway were performed. Besides the (Z),(Z)-octadeca-6,9-dienedioic acid, (Z),(Z)-3-hydroxyoctadeca-9,12-dienedioic acid and (Z),(Z)-3-hydroxytetradeca-5,8-dienedioic acid were obtained as the main fermentation products. The maximum concentrations of (Z),(Z)-octadeca-6,9-dienedioic acid and (Z),(Z)-3-hydroxyoctadeca-9,12-dienedioic acid reached values of 6.4 g/l and 6.9 g/l respectively. The structures of the products were characterized by chemical and spectroscopic methods. The configuration of the double bonds was not changed during bioconversion. As only one regioisomer of the hydroxylated fatty acid was detected, the hydroxylation is site-specific. Received: 11 November 1996 / Received revision: 11 February 1997 / Accepted: 24 February 1997     

Development of the male scent organ ofCreatonotos transiens (Lepidoptera,Arctiidae) during metamorphosis

Summary (1) The male abdominal scent organ (corema) of the arctiid mothCreatonotos transiens consists of a basal bladder and four tubes. It can be everted from the sternal intersegmental membrane 7/8. Its scent hairs (scales) produce and release the pheromone hydroxydanaidal, which attracts both sexes. Pyrrolizidine alkaloids (PA) ingested by the larva with its food are not only precursors of the pheromone but also a morphogen, which quantitatively controls the growth of the pupal corema and, thus, its final size and number of hairs. (2) The coremata arise from epidermalanlagen at the anterior border of the 8th abdominal sternite. If male larvae are fed 1 mg PA these organs begin to develop from small vesicles, and four tubes then arise during the first 3 pupal days. The corresponding mitoses reach their peak at 36 h. During the next 2 days the tubes shorten, while the walls become thin and doubly folded. The total surface of the corema increases about 20 times because of the shape transformation of the epidermal cells from prismatic to very flat. (3) The scent hairs originate from trichogen cells, which arise together with their associated tormogen cells during the 1st pupal day by way of differential mitoses. As the trichogen cells grow, their nuclei enlarge by way of endomitoses, elongate distally, and thus produce the hairs that extend into the lumen of the corema. Tormogen cells degenerate by the 8th day at latest. The hairs in each tube form a thick, caudally oriented bundle. The hair cells are finally bottle-shaped and at day 6 they extend freely into the hemolymph space. They are probably also the pheromoneproducing cells in later pupal and early imaginal life. Mitoses that produce trichogen cells stop after the 1st day, those producing epithelial cells 2 days later. This delay shifts the ratio of the two cell types from about 111 (18 h) to 140. (4) The processes hitherto described refer to normogenesis with ample PA supply. Control coremata in PA-free or PA-deficient specimens develop in principle in the same way, but at a slower rate, with minimal hair cell numbers barely 1/10th of normal, or at any rates between, depending upon the earlier PA supply. The size of control coremata varies from very small to small; even the hair cells and the hairs are smaller. (5) PA regulates corema development quantitatively through the number of mitoses of its cells and of endomitotic steps of the hair cells. In PA-treated specimens the coremaanlage is already advanced prior to pupation, at about the time when its sensitivity to PA influence terminates, in the early prepupa. Since PA only affects the anlagen of the corema and not that of any other body part (not even the basal coremal bladder), we postulate a selective interaction of PA with the presumptive corema cells. We found earlier that ecdysone is also involved, since the respective cell numbers can only be realized if this hormone is present.     

Preparation of Four Geometric Isomers of 4,6-Hexadecadienol, -dienal,and -dienyl Acetate and Their Electro-Antennographic Activity toward Male Eri-Silk Moths

Four geometric isomers of 4,6-hexadecadienol and the corresponding -dienal and -dienyl acetate isomers were prepared in order to evaluate their pheromone activity toward male eri-silk moths. We used the electroantennography (EAG)–GLC method to determine EAG activity, with results showing that only 4E,6Z- and 4Z,6E-hexadecadienal were active, while the other compounds were found to be totally inactive.     

棉铃虫成虫对两种关键花香气味的联系性学习

为了明确棉铃虫对苯乙醛和乙酸苯甲酯两种关键花香气味的联系性学习行为,在室内分别测定了不同学习训练的棉铃虫雌雄成虫对两种关键花香气味的选择偏好性反应和雄成虫的触角电位反应。偏好性测定结果表明,无花香气味接触经历的棉铃虫对苯乙醛和乙酸苯甲酯的选择频次均无显著差异,且单纯的花香气味接触经历也不能诱导棉铃虫气味偏好性的显著改变,唯有花香气味接触经历伴随蔗糖溶液进行强化训练时,才能诱导气味偏好性的显著性改变。从对两种花香气味的联系性学习看,棉铃虫更容易将苯乙醛与蔗糖溶液进行联系性学习,而不容易将乙酸苯甲酯与蔗糖溶液进行联系性学习。从性别差异看,雌虫比雄虫具有更强的将花香气味与蔗糖溶液进行联系性学习的能力。触角电位测定结果表明,不同学习训练的棉铃虫雄成虫对两种花香气味和绿叶气味顺-3-己烯-1-醇的EAG反应值之间不存在显著性差异。总之,花香气味伴随食物资源的联系性学习经历,能够明显提高棉铃虫成虫对花香气味的选择偏好,可以此为基础进一步优化花香引诱剂的配方设计和应用技术。     

Identification and field testing of sex pheromone components of a Korean population of the allium leafminer,Acrolepiopsis sapporensis

The allium leafminer, Acrolepiopsis sapporensis Matsumura (Lepidoptera: Acrolepiidae), is a pest of Allium species (Liliaceae) in Asia and Hawaii, USA. We identified candidate sex pheromone components in pheromone gland extracts of female moths and field tested the response of male moths to blends with different components and ratios. Gas chromatographic comparison of abdominal tip extracts from both sexes showed three female‐specific components: (Z)‐11‐hexadecenal (Z11‐16:Ald), (Z)‐11‐hexadecenyl acetate (Z11‐16:OAc), and (Z)‐11‐hexadecen‐1‐ol (Z11‐16:OH). These compounds were identified by mass spectral analysis of natural pheromone components and dimethyldisulfide adducts, and retention index comparisons with synthetic standards. The average ratio of three components, Z11‐16:Ald, Z11‐16:OAc, and Z11‐16:OH, in female extract was 33:100:14. Field trapping experiments indicated that all three components were essential for maximal attraction of male moths. Traps baited with a ternary blend mimicking the blend found in the pheromone gland extracts caught significantly more males than traps baited with caged live females. Increasing doses of the pheromone blend in the lures from 0.01 to 1.0 mg increased catches of male A. sapporensis.     

Female sex pheromone of a lichen moth Eilema japonica (Arctiidae, Lithosiinae): Components and control of production

Seven candidates for components of the female sex pheromone of Eilema japonica (Arctiidae, Lithosiinae) were detected in an extract of pheromone glands with a gas chromatograph-electroantennographic detector. The compounds were identified as (Z,Z)-6,9-icosadiene (D20), (Z,Z)-6,9-henicosadiene (D21), (Z,Z,Z)-3,6,9-henicosatriene (T21), (Z,Z)-6,9-docosadiene (D22), (Z,Z,Z)-3,6,9-docosatriene (T22), (Z,Z)-6,9-tricosadiene (D23), and (Z,Z,Z)-3,6,9-tricosatriene (T23). Assays using synthetic lures in a wind tunnel showed that D21 (proportion, 0.39), T21 (0.08), D22 (0.27), and T22 (0.26) are important for evoking full behavioral responses from the males. Titers of the pheromone components did not show clear temporal fluctuations. Moreover, decapitation of the female moth had no effect on the titers of pheromone components in the pheromone gland, suggesting that cephalic endocrine factors such as pheromone biosynthesis activating neuropeptide (PBAN) are not involved in the control of pheromone biosynthesis in this species.     

Electrophysiological and Behavioral Responses of Female Beet Armyworm Spodoptera exigua (Hübner) to the Conspecific Female Sex Pheromone

Electroantennogram (EAG) recordings showed that female Spodoptera exigua can detect their own sex pheromones (two single components and their mixture), displaying a similar dose–response pattern to that of males, although intensities of female responses were much less at all doses compared with males. Furthermore, the female calling behavior was inhibited and late-shifted by the presence of the female sex pheromone. When the pheromone components were presented, the calling female proportion in the peak calling period was significantly reduced and the calling peak time and calling termination time postponed, compared with controls. Although the calling behavior was inhibited, the pheromone titer of treated females was not different to the control, implying a reduced pheromone biosynthesis in the pheromone glands of treated moths. However, observations during the olfactometer experiments revealed that there were no obvious behavioral responses of females exposed to sex pheromone stimuli including whole gland extracts, 0.1, 1 or 10 μg binary pheromone mixtures.     

Spatial discrimination of pheromones and behavioural antagonists by the tortricid moths Cydia pomonella and Adoxophyes orana

Male moths responding to their species-specific sex pheromone, may cease their upwind flight when pheromone components of sympatric species are added to the mixture. The interspecific interaction between the pheromone response of the tortricid moths Cydia pomonella and Adoxophyes orana was investigated in field-trapping and wind-tunnel studies. Addition of the A. orana pheromone [(Z9)-tetradecenylacetate and (Z11)-tetradecenylacetate] to a source containing the C. pomonella pheromone [(E8, E10)-dodecadienol] resulted in a significant inhibition of attraction by male C. pomonella. It is demonstrated that this behavioural antagonist for C. pomonella must be emitted from the same point source to induce this inhibitory effect. A spatial separation of the two interspecific pheromones (at 14 cm, 5 cm and 0.5 cm crosswind) restored the attraction of the conspecific pheromone for male C. pomonella. In contrast to C. pomonella, male A. orana were not inhibited by point sources releasing both the C. pomonella and A. orana pheromone. We suggest that the discrepancy in the interspecific pheromone interaction between these two tortricids can be explained if we consider the evolutionary ecology of interspecific pheromone communication in C. pomonella and A. orana. Accepted: 18 July 1999     

Octopamine modulates the sensitivity of silkmoth pheromone receptor neurons

Effects of octopamine and its antagonist epinastine on electrophysiological responses of receptor neurons of Antheraea polyphemus specialised to the pheromone components (E,Z)-6,11-hexadecadienyl acetate and (E,Z)-6,11-hexadecadienal were investigated. Injections of octopamine and epinastine into the moths had no effect on the transepithelial potential of the antennal-branch preparation nor on the spontaneous nerve impulse frequency in either type of receptor neuron. However, in the presence of continuous low-intensity pheromone stimulation, octopamine significantly increased the nerve impulse frequency in the acetate receptor neuron, but not in the aldehyde receptor neuron. Octopamine and epinastine had no significant effect on the receptor potential amplitudes elicited in both receptor neuron types by pheromone stimulation. However, the peak nerve impulse frequency in the response of both receptor neuron types to pheromone was significantly affected: decreased by epinastine and increased by octopamine over a broad range of pheromone concentrations. In control experiments, injection of physiological saline did not significantly alter the peak nerve impulse frequency. The effect of octopamine was established within 1 h after injection and persisted for about 4 h. The possibility of a direct action of octopamine on the nerve impulse generation by the receptor neurons is discussed. Accepted: 8 January 2000     

Detection of short pure-tone stimuli in the noctuid ear: what are temporal integration and integration time all about?

The interception of a pheromone filament induces flying moths to surge briefly nearly straight upwind; in the absence of pheromone moths cease upwind progress and zigzag crosswind. We tested males of the almond moth, Cadra cautella (Lepidoptera, Pyralidae), in a low-turbulence wind tunnel in wind velocities of 20, 40 and 80 cm s⁻¹. A mechanical pulse generator was set to produce plumes either with same pheromone pulse frequency (pulse generation frequency of 2.9 Hz, interpulse distances from 7 cm to 28 cm) or plumes with same interpulse distance across the three wind velocities (interpulse distance of 14 ± 2 cm, pulse generation frequency of 1.7–5.0 Hz). In plumes of similar pulse frequency, the faster the speed of the wind the slower the ground speed of flight. However, in plumes of similar interpulse distance, ground speed remained relatively constant independent of the wind speed. A `realized' frequency of pulse interception for males flying along the various combinations of pulse frequencies and wind velocities was calculated using the males' average airspeed and the spatial distribution of pheromone pulses in the plume. Realized frequency of pulse interception ranged from 1.3- to 3.0-fold higher than the frequency of pulse generation. The flight tracks of males reflected the regime of realized pulse interception. These results suggest that upwind flight orientation of male C. cautella to pheromone in different wind velocities is determined by the flux of filament encounter. Accepted: 3 September 1997     

Fine-scale resolution of closely spaced pheromone and antagonist filaments by flying male Helicoverpa zea

The limits of a male moth's ability to resolve closely spaced odor filaments have been investigated. Male Helicoverpa zea normally respond to their conspecific sex pheromone blend by exhibiting an upwind flight, which culminates in source contact by at least 50% of the bioassayed individuals. When loaded onto the same filter paper source containing this hitherto attractive pheromone blend, or onto a separate filter paper and co-emitted from the same pipette source with pheromone, (Z)-11-hexadecenyl acetate severely reduced upwind flight and source contact by male H. zea. A similar level of upwind flight inhibition was recorded when the antagonist (Z)-11-hexadecenyl acetate was emitted from its own point source placed 1 mm upwind of the pheromone point source, both plumes being simultaneously emitted in a continuous mode to form a confluent strand. However, (Z)-11-hexadecenyl acetate was less effective in reducing upwind flight and source contact when it was isolated and pulsed from its own source, placed 1 mm either upwind, downwind or cross-wind of a pipette source from which pheromone was simultaneously being pulsed, such that both filaments were separated in time by 0.001–0. 003 s. These results suggest that male H. zea are able to distinguish between odor sources separated by as little as 1 mm in space and 0.001 s in time. Accepted: 31 March 1999     

Responses of olfactory receptor neurons in Utetheisa ornatrix to gender-specific odors

We recorded the electrophysiological responses of individual olfactory receptor neurons in sensilla trichodea on the antennae of adult arctiid moths, Utetheisa ornatrix, to stimulation with volatiles associated with both sexes. All trichoid sensilla contain at least two receptor neurons, each with distinct action potential amplitudes and waveforms, that respond dichotomously to male and female odors. Although, neither female neuron responds to extracts of coremata or the male-produced pheromone hydroxydanaidal, they do respond in a gender-specific manner to the volatiles emanating from whole pupae, hemolymph, thoracic froth, and adult animals of several ages. Thoracic froth, which contains pyrrolizidine alkaloids, is thought to play a role in defense. Froth from moths reared on diets, with or without added pyrrolizidine alkaloids, were equally effective in eliciting gender-specific patterns of response. Male trichoid receptor neurons respond to these same materials with similar patterns of activation. These receptor neurons provide information about substances, which we have termed “gender odors,” that are persistently emitted by nearby animals. These substances do not appear to be the same as those already known to be involved in defense or the sexual dialog between individuals. Accepted: 16 March 2000     

The egg parasitoid <Emphasis Type="Italic">Telenomus euproctidis</Emphasis> (Hymenoptera: Scelionidae) uses sex pheromone released by immobile female tussock moth <Emphasis Type="Italic">Orgyia postica</Emphasis> (Lepidoptera: Lymantriidae) as kairomone

Egg parasitoids Telenomus euproctidis Wilcox (Hymenoptera: Scelionidae) were attracted to egg masses laid by wingless immobile female Orgyia postica (Lepidoptera: Lymantriidae). Virgin females, a solvent extract of pheromone glands, and a synthetic sex pheromone, (6Z,9Z,11S,12S)-11,12-epoxyhenicosa-6,9-diene (posticlure), also attracted this parasitoid in the field, demonstrating that T. euproctidis uses the sex pheromone of female Org. postica as a kairomone to locate host eggs.     

A Selective Method of Estimation of Bacitracin in the Presence of Other Antibiotics and Growth Factors

The plum cankerworm moth, Cystidia couaggaria couaggaria (Geometridae: Ennominae), is a defoliator of Chinese plum trees (Prunus mume). The pheromone components of the female were analyzed by gas chromatography (GC) with an electro-antennographic (EAG) detector and GC coupled with mass spectrometry. The crude pheromone extract included several EAG-active components, i.e., trienyl, dienyl, and saturated hydrocarbons, with a C₂₁–C₂₅ straight chain. The characteristic mass spectra indicated the unsaturated hydrocarbons to be (3Z,6Z,9Z)-3,6,9-trienes and (6Z,9Z)-6,9-dienes. In the fields, mixtures of the synthetic C₂₁ and C₂₃ trienes in a ratio of 2:3 and 1:4 successfully attracted males of this diurnal species during daytime. While the male antennae responded to the C₂₅ triene and saturated hydrocarbons, their synergistic effects were not observed on the male attraction in the fields. Addition of the C₂₁ diene interestingly inhibited the activity of the triene mixture. Males of Cystidia truncangulata, a sympatric diurnal congener of C. c. couaggaria, showed similar EAG responses to the unsaturated hydrocarbons, but no C. truncangulata males were attracted by the lures tested for C. c. couaggaria males, indicating that the identified hydrocarbons comprised the species-specific pheromone of C. c. couaggaria females.     

Antennal responses of Cydia pomonella (L.) exposed to surfaces treated with methoxyfenozide

Electroantennogram (EAG) measurements were recorded from the antennae of male and female codling moth, Cydia pomonella L., to determine whether adult moths exposed to surfaces treated with the ecdysteroid agonist methoxyfenozide experience a decline in their antennal reception and thus olfactory sensitivity. Such a phenomenon would offer a possible mechanism for the previously reported decreased responsiveness from moths treated with methoxyfenozide to pheromone‐ and plant volatile‐based monitoring lures. Mean EAG data revealed that the antennae from methoxyfenozide‐treated male moths appear to be just as sensitive to various doses of synthetic codlemone as the antennae from the control and surfactant‐treated moths, but they appeared to be less sensitive to the pheromone component 12OH (collected from female effluvia) than the control male antennae. Mean male EAG responses to the pheromone components E8,E10‐12Al and codlemone collected from methoxyfenozide‐treated females were significantly less than the responses towards those two pheromone components collected from the control and surfactant‐treated females. Female moth exposure to methoxyfenozide did not negatively impact the sensitivity of female antennae to the plant volatile pear ester, but it did towards the apple volatile butyl hexanoate. Data from this study show that adult C. pomonella exposure to methoxyfenozide‐treated surfaces does appear to negatively impact, in a minor way, the (i) olfactory sensitivity (or detection) of male antennae towards some components of the female sex pheromone, (ii) the female antennal sensitivity towards a key apple volatile and (iii) the attractiveness of female pheromone effluvia.     

Olfactory attraction to aggregation pheromone is mediated by disti-flagellum of antennal segments in Riptortus pedestris

Riptortus pedestris (Hemiptera: Alydidae) is a serious pest of soybean and sweet persimmon and uses male produced aggregation pheromone, (E)-2-hexenyl (Z)-3-hexenoate, (E)-2-hexenyl (E)-2-hexenoate, and tetradecyl isobutyrate to facilitate food location and recognition by conspecifics. Using electroantennogram (EAG) and greenhouse bioassay, we determined which antennal segment is involved in the detection of their aggregation pheromone. In the first EAG test using individual antennal segments, significant EAG responses to 1:1:1 mixture of the aggregation pheromone were observed only from the disti-flagellum segments of both male and female antennae at both pheromone doses tested (1 µg and 100 µg). In the following EAG tests using gradually removed antennal segment(s), EAG response was still maintained when the distal half of a disti-flagellum was surgically removed, while EAG response was lost when whole segment of disti-flagellum or other whole segments were gradually removed from intact antenna of both sexes. In greenhouse experiment, removing one or both segment(s) of disti-flagellum from male or female antennae resulted in significant reduction in their attraction to the aggregation pheromone. Together, these findings support that the disti-flagellum of R. pedestris houses olfactory neurons associated with attraction to their aggregation pheromone.     

Sex pheromone composition of the variegated cutworm,Peridroma saucia (Lepidoptera: Noctuidae), in Korea

This study was conducted to investigate the sex pheromone composition of the variegated cutworm (Peridroma saucia Hübner) in Korea. The sex pheromone components of P. saucia were identified as (Z)-9-tetradecenyl acetate (Z9-14:Ac) and (Z)-11-hexadecenyl acetate (Z11-16:Ac) through GC-EAD and GC–MS analysis. EAG tests of the male antennae revealed that the Z9-14:AC exerted significantly larger responses than other compounds. The female moths primarily called and copulated between 6 h and 7 h after the lights off, and the ratio of two pheromone components, Z9-14:Ac and Z11-16:Ac, in the sex pheromone gland during this period was 1:2.1 to 1:2.4. In the field trapping studies, a large number of male moths were caught in the traps baited with the mixtures of Z9-14:Ac and Z11-16:Ac at the ratios ranging from 2.3:1 to 1:4, with the highest trap catches at 1:1 to 1:2.3 ratios of the two components. The seasonal flight activities of P. saucia monitored by using pheromone lures revealed complicated patterns in Korea. Specifically, the first flight period was spread over a long period and irregular, while the second flight period differed among the localities examined.     

Pyrrolizidine alkaloids quantitatively regulate both scent organ morphogenesis and pheromone biosynthesis in maleCreatonotos moths (Lepidoptera: Arctiidae)

Summary Males ofCreatonotos gangis andC. transiens possess coremata (scent organs) of drastically varying sizes (Figs. 2, 3), which release R(-)-hydroxydanaidal (Fig. 1A) in varying amounts. Both the size of the organs and their pheromone content depend on the ingestion of pyrrolizidine alkaloids (PAs; Fig. 1 B, C) by the larvae. There is a direct correlation between amounts of PAs ingested and the size of the organs (Fig. 4). It is the absolute amount of PAs ingested which determines the expression of corematal size, structurally different PAs have identical effects (Table 2); PAs are no essential dietary factors for the general development of the moths (Table 1), and the morphogenetic effect is restricted to the coremata. The findings are discussed in terms of developmental, ecological and functional aspects.Abbreviation PAs pyrrolizidine alkaloids