Sex Pheromone Composition Research Articles

Evolution of moth sex pheromone composition by a single amino acid substitution in a fatty acid desaturase

For sexual communication, moths primarily use blends of fatty acid derivatives containing one or more double bonds in various positions and configurations, called sex pheromones (SPs). To study the molecular basis of novel SP component (SPC) acquisition, we used the tobacco hornworm (Manduca sexta), which uses a blend of mono-, di-, and uncommon triunsaturated fatty acid (3UFA) derivatives as SP. We identified pheromone-biosynthetic fatty acid desaturases (FADs) MsexD3, MsexD5, and MsexD6 abundantly expressed in the M. sexta female pheromone gland. Their functional characterization and in vivo application of FAD substrates indicated that MsexD3 and MsexD5 biosynthesize 3UFAs via E/Z14 desaturation from diunsaturated fatty acids produced by previously characterized Z11-desaturase/conjugase MsexD2. Site-directed mutagenesis of sequentially highly similar MsexD3 and MsexD2 demonstrated that swapping of a single amino acid in the fatty acyl substrate binding tunnel introduces E/Z14-desaturase specificity to mutated MsexD2. Reconstruction of FAD gene phylogeny indicates that MsexD3 was recruited for biosynthesis of 3UFA SPCs in M. sexta lineage via gene duplication and neofunctionalization, whereas MsexD5 representing an alternative 3UFA-producing FAD has been acquired via activation of a presumably inactive ancestral MsexD5. Our results demonstrate that a change as small as a single amino acid substitution in a FAD enzyme might result in the acquisition of new SP compounds.

Artificial night lighting disrupts sex pheromone in a noctuid moth

1. One major, yet poorly studied, change in the environment is the increase in nocturnal light pollution. Although this strongly alters the habitat of nocturnal species, the ecological consequences are poorly known. Moths are well known to be attracted to artificial light sources, but artificial light may affect them in other ways as well.2. In this study, female Mamestra brassicae moths were subjected to various types of low‐intensity artificial night lighting with contrasting spectral compositions (green‐rich, red‐rich, warm white) or to a dark control treatment and the effects on their sex pheromone production and composition were tested.3. Artificial night lighting reduced sex pheromone production and altered the chemical composition of the pheromone blend, irrespective of spectral composition. Specifically, amounts of the main pheromone component Z11‐16:Ac were reduced, while the deterring compounds Z9‐14:Ac, Z9‐16:Ac, and Z11‐16:OH were increased relative to Z11‐16:Ac when females were kept under artificial light. These changes may reduce the effectiveness of the sex pheromones, becoming less attractive for males.4. These results show for the first time that artificial light at night affects processes that are involved in moth reproduction. The potential for mitigation through manipulation of the spectral composition of artificial light appears limited.

Selection on male sex pheromone composition contributes to butterfly reproductive isolation.

Selection can facilitate diversification by inducing character displacement in mate choice traits that reduce the probability of maladaptive mating between lineages. Although reproductive character displacement (RCD) has been demonstrated in two-taxa case studies, the frequency of this process in nature is still debated. Moreover, studies have focused primarily on visual and acoustic traits, despite the fact that chemical communication is probably the most common means of species recognition. Here, we showed in a large, mostly sympatric, butterfly genus, a strong pattern of recurrent RCD for predicted male sex pheromone composition, but not for visual mate choice traits. Our results suggest that RCD is not anecdotal, and that selection for divergence in male sex pheromone composition contributed to reproductive isolation within the Bicyclus genus. We propose that selection may target olfactory mate choice traits as a more common sensory modality to ensure reproductive isolation among diverging lineages than previously envisaged.

Intraspecific Variation in Female Sex Pheromone of the Codling Moth Cydia pomonella.

The codling moth, Cydia pomonella L. (Lepidoptera, Tortricidae), is a major pest of apple, pear and walnut orchards worldwide. This pest is often controlled using the biologically friendly control method known as pheromone-based mating disruption. Mating disruption likely exerts selection on the sexual communication system of codling moth, as male and female moths will persist in their attempt to meet and mate. Surprisingly little is known on the intraspecific variation of sexual communication in this species. We started an investigation to determine the level of individual variation in the female sex pheromone composition of this moth and whether variation among different populations might be correlated with use of mating disruption against those populations. By extracting pheromone glands of individual females from a laboratory population in Canada and from populations from apple orchards in Spain and Italy, we found significant between- and within-population variation. Comparing females that had been exposed to mating disruption, or not, revealed a significant difference in sex pheromone composition for two of the minor components. Overall, the intraspecific variation observed shows the potential for a shift in female sexual signal when selection pressure is high, as is the case with continuous use of mating disruption.

Evidence for divergence in cuticular hydrocarbon sex pheromone between California and Mississippi (United States of America) populations of bark beetle parasitoid Roptrocerus xylophagorum (Hymenoptera: Pteromalidae)

AbstractRoptrocerus xylophagorum (Ratzeburg) (Hymenoptera: Pteromalidae) is a common Holarctic parasitoid of the larvae and pupae of bark beetles (Coleoptera: Curculionidae: Scotytinae). In no-choice laboratory bioassays, we found that male wasps derived either from northern California or southwestern Mississippi, United States of America more frequently displayed sexual behaviours (including mounting, wing fanning, and copulation attempts) to glass bulb decoys treated with hexane cuticular washes of females derived from the same parasitoid population rather than the distant population. This result suggests that the composition of the cuticular hydrocarbon sex pheromone has diverged between eastern and western populations and is consistent with previous data indicating that R. xylophagorum may consist of more than one species.

Identification and application of the sex pheromones of the rice green caterpillar Naranga aenescens in Korea

This study was carried out to investigate the optimum composition of sex pheromones for mate attraction in the rice green caterpillar Naranga aenescens, and to evaluate the effects of pheromone quantity, trap type, and trapping location on trapping success in Korean paddy fields. Mating rate of N. aenescens increased with age, with the highest mating rate 2days after emergence. The highest mating rate occurred within 1h after lights-off; mating pairs were not observed during the photophase. In electroantennogram (EAG) and gas chromatography with electroantennographic detection (GC–EAD) analysis, virgin female extracts had 2 EAD-active components. The mass spectra of the 2 distinct EAD-active components were identical to those of Z9-14:Ac and Z11-16:Ac. Additionally, Z9-16:Ac was identified in the extracts in gas chromatography–mass spectrometry (GC–MS) analysis. Wing traps with lures containing 1mg of a 2:1:4 mixture of Z9-14:Ac, Z9-16:Ac, and Z11-16:Ac was the most attractive to N. aenescens males. Pheromone-baited wing trapping, therefore, can be a simple and effective tool for monitoring N. aenescens populations.

복숭아명나방에 대한 페로몬 트랩의 처리조건에 따른 유인효과

Abstract Mating disruption experiments were evaluated and compared in several pheromone traps and sexpheromone composition ratios at commercial chestnut garden for developing alternative control method of D.punctiferalis. The highest attractive was shown in Delta traps at the effective sex pheromone ratio of 75:25(E10-16:A1:Z10-16:A1). Comparisons of domestic and imported pheromone constituents were not significantdifference in 90:10 (E10-16:A1:Z10-16:A1). But, there was significant difference in sex pheromone ratio of75:25. D. punctiferalis was caught the highest number in Gongju area for attractive test using pheromonetrap. Attractive effects with dose of pheromone were showed the highest at 2.5 mg/lure and 1 mg/lure inGongju area and Cheongyang area respectively. However, there was no significant difference amongpheromone dose in Buyeo area. These data could be useful for eco-friendly managing of D. punctiferalis inthe chestnut farm.Key words Delta trap, Dichocrocis punctiferalis, (E)-10-hexadecenyl aldehyde, Sex pheromone, (Z)-10-hexa-decenyl aldehyde

Effects of different sex pheromone compositions and host plants on the mating behavior of two Grapholita species

The two congener species Grapholita molesta and Grapholita dimorpha share two major sex pheromone components: cis-8-dodecenyl acetate (Z8-12Ac) and trans-8-dodecenyl acetate (E8-12Ac). In fact, commercial sex pheromone lures composed of only these two major components attract the males of both species. In this study, we aimed to determine the reproductive isolation components of these two species by analyzing the effects of the minor sex pheromone components and host plants. First, different ratios of the two major sex pheromone components were greatly favored by either male species. Sex pheromone gland extracts of G. dimorpha contained a lesser proportion of Z8-12Ac than that of G. molesta. In the three (apple, pear, and peach) orchards investigated in this study, a larger number of G. molesta males were attracted to the 95:5 pheromone mixture (Z8-12Ac and E8-12Ac, respectively), while a larger number of G. dimorpha males were attracted to the 85:15 mixture. Second, there was a significant variation in male attractions in different host plants. G. molesta males were more attracted to the sex pheromone lure in the apple orchards than that in the pear and peach orchards. In contrast, G. dimorpha males were more attracted to the lures in the pear and peach orchards than that in the apple orchard. Third, the minor sex pheromone components were important for reproductive isolation. Among the four minor components tested, addition of (Z)-8-dodecenol (Z8-12OH) to the major sex pheromone components significantly suppressed male attraction in G. dimorpha and slightly elevated male attraction in G. molesta. The discriminating effect of Z8-12OH was further validated using male electroantennogram analysis. These results suggest that reproductive isolation between two congeners can be achieved by variations in the minor sex pheromone components and in the host plants, as well as by changes in the ratio of the two major components.

국내 배과원에 복숭아순나방붙이의 발생과 유사종 복숭아순나방의 페로몬 트랩 교차 유인

최근 사과원에서 복숭아순나방붙이(Grapholita dimorpha)의 발생이 보고되었다. 복숭아순나방붙이는 이와 유사한 복숭아순나방(G. molesta)이 발생하는 또 다른 기주에서도 동시에 발생이 가능하다고 제기되었다. 본 연구는 국내 여러 지역의 배과원에서 복숭아순나방붙이의 발생이 있음을 보고한다. 복숭아순나방붙이의 종 동정은 형태적 특징과 분자마커를 이용하여 실시되었다. 이들 두 종의 공통된 성페로몬 주성분으로 상호 교차 포획이 이뤄질 수 있다. 복숭아순나방붙이 페로몬트랩에 복숭아순나방붙이와 복숭아순나방이 포획되고, 복숭아순나방 페로몬트랩에 복숭아순나방과 복숭아순나방붙이가 포획되었다. 복숭아순나방붙이 트랩에 이뤄진 교차 포획비율은 지역적으로 다른 배과원에서 상호 뚜렷한 차이가 나타났다. 더욱이 이 두 종의 발생 피크도 조사한 모든 야외 지역에서 시기적으로 뚜렷한 차이를 보였다. 이러한 결과는 배 과수원에서 두 종의 성페로몬 트랩으로 얻은 모니터링 자료는 각각 서로 다른 종의 포획이 혼재하며, 이는 해당종의 발생빈도와 발생밀도가 확대 해석될 수 있음을 본 연구 자료는 제시하고 있다. The occurrence of plum fruit moth, Grapholita dimorpha, has been recently reported in apple orchards in Korea. It has been suspected that G. dimorpha and its related species, G. molesta, may occur simultaneously in other host plants. This study reports the occurrence of G. dimorpha in pear orchards of different localities in Korea. The identification of G. dimorpha was determined by morphological characters and the DNA marker. The cross-trapping of both species may be possible because the major sex pheromone (SP) compositions for the two species are similar. From the monitoring data, G. dimorpha and G. molesta were caught in SP lure traps of G. dimorpha and both species were also caught in SP lure traps of G. molesta. This cross-trapping of G. molesta to a SP lure of G. dimorpha varied significantly among pear orchards in different geographical localities. Furthermore, the occurrence peaks of the two species were not coincidental in all monitored orchards. These suggest that monitoring data obtained from each SP trap of both species in pear orchards may be mixed with two species, which would result in the over-estimation of population density and peak frequency on both species in pear orchards.

Cracking the olfactory code of a butterfly: the scent of ageing

Although olfaction is a primary mode of communication, its importance in sexual selection remains understudied. Here, using the butterfly Bicyclus anynana, we address all the parameters of importance to sexual selection for a male olfactory signal. We show that variation in the male sex pheromone composition indicates male identity and male age. Courting males of different ages display small absolute (c. 200 ng) but large relative (100%) change of one specific pheromone component (hexadecanal) which, unlike the other components, showed no heritability. Females prefer to mate with mid-aged over younger males and the pheromone composition is sufficient to determine this preference. Surprisingly refined information is thus present in the male olfactory signal and is used for sexual selection. Our data also reveal that there may be no 'lek paradox' to resolve once the precise signal of importance to females is identified, as hexadecanal is, as expected, depleted in additive genetic variation.

Sex pheromone composition of Ascotis selenaria (Lepidoptera: Geometridae) and its regional variation in Korea

This study was conducted to investigate sex pheromone composition of Ascotis selenaria (Lepidoptera: Geometridae) in Korea. Two sex pheromone compounds such as (Z,Z)-6,9-cis-3,4-epoxynonadecadiene (6Z,9Z-cis-3,4-epoxy-19:H) and (Z,Z,Z)-3,6,9-nonadecatriene (3Z,6Z,9Z-19:H) were identified in the glands of A. selenaria females by gas chromatography–mass spectrometry analysis. However, the component 3Z,6Z,9Z-19:H neither elicited an electroantennogram response nor increased the attractiveness for A. selenaria males in the field. The role of 3Z,6Z,9Z-19:H seems to be as an antagonistic signal for mating behavior of A. selenaria males.The blend ratios of two 6Z,9Z-cis-3,4-epoxy-19:H isomers such as, 6Z,9Z-cis-3R,4S-epoxy-19:H and 6Z,9Z-cis-3S,4R-epoxy-19:H, were critical to attract A. selenaria males. The blend ratios of the two isomers showing peak catch of A. selenaria males had large variations among the locations investigated. A. selenaria populations in Gunwi showed peak activity at ratios of 0.9:0.1 and 0.8:0.2, whereas the populations in Goheung, Yeongam, and Jeju (Aewol and Harye) showed peak activity at a 0.5:0.5 ratio. In Changnyeong, the peak activity occurred in a bimodal form at ratios of 0.7:0.3 and 0.4:0.6. Such variation was partially explained by geographical isolation due to mountain ranges. Consequently, the results of our study should be useful for designing a region-specific pheromone lure for successful A. selenaria monitoring.

Sex pheromone composition and reproductive isolation in two Busseola species (Lepidoptera: Noctuidae) in Kenya

Closely related species provide an interesting and useful model to understand mate communication diversification. Pre-mating isolation is the result of selection processes and is achieved by utilization of different sex pheromones, but also by different ecological and life traits such as geographic distribution, host-plant specialization and allochrony of sexual behavior. Here, we report sex pheromone identifications and pre-mating isolation of two closely related species of stemborers: Busseola segeta and the yet undescribed Busseola nov. sp., developing on different host-plants and in different geographical locations in Kenya. GC–MS analyses showed that the two species produced the same pheromone components, but in slight different ratios. The respective synthetic pheromone mixtures tested in the field were attractive for males of each species. The low inter-specific variation in the pheromone mixture would allow possible interbreeding in case of accidental introduction of both species in the same environment. Under laboratory conditions, allochrony of the mating period is total and might prevent interbreeding if confirmed in natural conditions. This point would ensure the reproductive isolation between the two species in case of modifications of ecological factors. This study and the chemical structures of the identified pheromone components clearly demonstrate that these Busseola species are closely related although they differ in morphology.

Solid Phase Microextraction of Volatile Emissions of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae): Influence of Fly Sex, Age, and Mating Status

Considerable efforts have been devoted to understanding the courtship behavior and pheromone communication of medflies; however, the sex pheromone composition is still a controversial subject. The discovery of new components affecting medfly behavior would be of interest for medfly control methods based on semiochemicals. This work describes volatile compounds emitted by Ceratitis capitata collected using solid phase microextraction. The volatile study was conducted according to an experimental design with three factors (sex, age, and mating status) assumed to be relevant for better understanding the chemical communication. Emission data were treated by means of principal component analysis, a statistical methodology not previously applied to the study of volatiles emitted by fruit flies. The characterization of emission patterns could be useful for the selection of compounds to be further investigated in biological assays to improve knowledge of the key semiochemicals involved in medfly behavior.

Variation in Sexual Communication of the Tobacco Budworm,Heliothis virescens

Abstract. Females of the tobacco budworm, Heliothis virescens (F.), exhibit distinct geographical and temporal variation in sex pheromone composition, but the causes and significance of this variation are largely unexplored. Here we assessed whether 1) female pheromone variation was related to the host plants of origin, and 2) pheromone lures with varying amounts of Z9–14:Ald or 16:Ald were differentially attractive to males. Variation in female pheromone did not seem to be related to the host plants from which the eggs or larvae were collected, which may be because field-collected larvae were reared for three to five larval stages on artificial diet. By varying the concentration of Z9–14:Ald within the range in the female pheromone gland, we found males were more attracted as the amount increased from 1 to 10% relative to Z11–16:Ald, but significantly less with the highest concentration of 25%. In contrast, with 16:Ald, similar numbers of tobacco budworm males were caught in all traps where 16:Ald ranged...

The roles and interactions of reproductive isolation mechanisms in fall armyworm (Lepidoptera: Noctuidae) host strains

1. The moth Spodoptera frugiperda presents an interesting opportunity to study the evolution of reproductive isolation, because it consists of two host races that may be in the process of speciation.2. The two races exhibit habitat isolation through host‐plant preference, and two types of behavioural isolation, i.e. differences in sex pheromone composition and timing of mating activity at night.3. In this paper, we review the selection pressures acting upon these three barriers as well as their genetic bases, to address the question of how divergence of the two strains may have evolved.4. We also address possible interactions between the three barriers, whether and how they may have evolved in concert, and we view the evolution of these three prezygotic isolation barriers in the light of postzygotic isolation.

Sex pheromone composition of the cotton caterpillar, Palpita indica (Lepidoptera: Pyralidae), in Korea

This study was conducted to investigate the sex pheromone composition of Palpita (= Diaphania) indica (Saunders) (Lepidoptera: Pyralidae) in Korea. Two sex pheromone components, E11-16:Al and E,E-10,12-16:Al, were identified by GC and GC-EAD analysis of sex pheromone gland extracts. The mean ratios of the two components, E11-16:Al and E,E-10,12-16:Al, were about 6.5:3.5 and 7.5:2.5 in gland extracts and in SPME collection, respectively. In field bioassays, maximum attraction occurred in traps baited with a 7:3 ratio of E11-16:Al and E,E-10,12-16:Al. The head extracts of P. indica stimulated the sex pheromone production of P. indica, as well as Hez-PBAN and PssPT, indicating that a PBAN-like substance exists in the head extracts of P. indica. Whole-mount immunocytochemistry showed that three groups of neurosecretory cells showed PBAN-like immunoreactivity in the SEG of P. indica. The PBAN-like immunoreactivity connected to the Corpora Cardiaca, a neurohemal organ. Also, PBAN-like immunoreactivity was found in the brain and in the thoracic and third/fourth abdominal ganglia. The addition of sex pheromone components of Peridroma saucia to the sex pheromone of P. indica greatly improved the attraction of P. indica males. The mixing of the sex pheromone components of S. exigua did not significantly increase the catches of P. indica, while the sex pheromone of H. armigera completely inhibited the attractiveness.

The evolution of sex pheromones in an ecologically diverse genus of flies

In theory, pheromones important in specific mate recognition should evolve via large shifts in composition (saltational changes) at speciation events. However, where other mechanisms exist to ensure reproductive isolation, no such selection for rapid divergence is expected. In Bactrocera fruit flies (Diptera: Tephritidae), males produce volatile chemicals to attract females for mating. Bactrocera species exhibit great ecological diversity, with a wide range of geographical locations and host plants used. They also have other mechanisms, including temporal and behavioural differences, which ensure reproductive isolation. Therefore, we predicted that their sex pheromones would not exhibit rapid divergence at speciation events. In the present study, we tested this idea by combining data on male sex pheromone composition for 19 species of Bactrocera with a phylogeny constructed from DNA sequence data. Analyses of the combined data revealed positive correlations between pheromone differences and nucleotide divergence between species, and between the number of pheromone changes along the phylogeny and the branch lengths associated with these changes. These results suggest a gradual rather than saltational mode of evolution. However, remarkable differences in sex pheromones composition exist, even between closely-related species. It appears therefore that the mode of evolution of sex pheromones in Bactrocera is best described by rapid saltational changes associated with speciation, followed by gradual divergence thereafter. Furthermore, species that do not overlap ecologically are just as different pheromonally as species that do. Thus, large changes in pheromone composition appear to be achieved, even in cases where other mechanisms to ensure reproductive isolation exist. We suggest that these differences are closely associated with rapid changes in host plant use, which is a characteristic feature of Bactrocera speciation.

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.

The Male Sex Pheromone of the Butterfly Bicyclus anynana: Towards an Evolutionary Analysis

BackgroundFemale sex pheromones attracting mating partners over long distances are a major determinant of reproductive isolation and speciation in Lepidoptera. Males can also produce sex pheromones but their study, particularly in butterflies, has received little attention. A detailed comparison of sex pheromones in male butterflies with those of female moths would reveal patterns of conservation versus novelty in the associated behaviours, biosynthetic pathways, compounds, scent-releasing structures and receiving systems. Here we assess whether the African butterfly Bicyclus anynana, for which genetic, genomic, phylogenetic, ecological and ethological tools are available, represents a relevant model to contribute to such comparative studies.Methodology/Principal FindingsUsing a multidisciplinary approach, we determined the chemical composition of the male sex pheromone (MSP) in the African butterfly B. anynana, and demonstrated its behavioural activity. First, we identified three compounds forming the presumptive MSP, namely (Z)-9-tetradecenol (Z9-14:OH), hexadecanal (16:Ald ) and 6,10,14-trimethylpentadecan-2-ol (6,10,14-trime-15-2-ol), and produced by the male secondary sexual structures, the androconia. Second, we described the male courtship sequence and found that males with artificially reduced amounts of MSP have a reduced mating success in semi-field conditions. Finally, we could restore the mating success of these males by perfuming them with the synthetic MSP.Conclusions/SignificanceThis study provides one of the first integrative analyses of a MSP in butterflies. The toolkit it has developed will enable the investigation of the type of information about male quality that is conveyed by the MSP in intraspecific communication. Interestingly, the chemical structure of B. anynana MSP is similar to some sex pheromones of female moths making a direct comparison of pheromone biosynthesis between male butterflies and female moths relevant to future research. Such a comparison will in turn contribute to understanding the evolution of sex pheromone production and reception in butterflies.

Genetic basis of sex pheromone blend difference between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

The two closely related moth species, Helicoverpa armigera and H. assulta, are sympatric in China. Both species use a mixture of (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-9-hexadecenal (Z9-16:Ald) as their sex pheromones but in widely different ratios. Hybridization and backcrossing experiments between H. armigera and H. assulta were conducted and sex pheromone compositions of the parent species, their F1 hybrids and backcrosses were compared to study the genetic basis of the production of their sex pheromone blend composition. Results show that the difference in sex pheromone blend ratios of these Helicoverpa species is mainly controlled by an autosomal locus with two alleles, with the allele from H. armigera being almost completely dominant over that derived from H. assulta.