The isolation and identification of the female sex pheromones of the red bollworm moth, Diparopsis castanea

The female sex pheromones of the red bollworm moth, Diparopsis castanea, were isolated by solvent extraction of the terminal abdominal segments of the female moth followed by purification of the solvent extract by liquid-solid and gas chromatography. The pheromones were identified by gas chromatographic analysis, micro-ozonolysis, infra-red and mass spectroscopy, and comparison with synthetic material. The major pheromone was 9,11-dodecadien-1-yl acetate (80 : 20 trans : cis). Minor components identified were trans 9-dodecen-1-yl acetate, 11-dodecen-1-yl acetate, and dodecan-1-yl acetate.     

More to legs than meets the eye: Presence and function of pheromone compounds on heliothine moth legs

Chemical communication is ubiquitous in nature and chemical signals convey species-specific messages. Despite their specificity, chemical signals may not be limited to only one function. Identifying alternative functions of chemical signals is key to understanding how chemical communication systems evolve. Here, we explored alternative functions of moth sex pheromone compounds. These chemicals are generally produced in, and emitted from, dedicated sex pheromone glands, but some have recently also been found on the insects' legs. We identified and quantified the chemicals in leg extracts of the three heliothine moth species Chloridea (Heliothis) virescens, Chloridea (Heliothis) subflexa and Helicoverpa armigera, compared their chemical profiles and explored the biological function of pheromone compounds on moth legs. Identical pheromone compounds were present on the legs in both sexes of all three species, with no striking interspecies or intersex differences. Surprisingly, we also found pheromone-related acetate esters in leg extracts of species that lack acetate esters in their female sex pheromone. When we assessed gene expression levels in the leg tissue, we found known and putative pheromone-biosynthesis genes expressed, which suggests that moth legs may be additional sites of pheromone production. To determine possible additional roles of the pheromone compounds on legs, we explored whether these may act as oviposition-deterring signals, which does not seem to be the case. However, when we tested whether these chemicals have antimicrobial properties, we found that two pheromone compounds (16:Ald and 16:OH) reduce bacterial growth. Such an additional function of previously identified pheromone compounds likely coincides with additional selection pressures and, thus, should be considered in scenarios on the evolution of these signals.     

Tissue localization and partial characterization of pheromone biosynthesis activating neuropeptide inAchaea janata

Female sex pheromone production in certain moth species have been shown to be regulated by a cephalic endocrine peptidic factor: pheromone biosynthesis activating neuropeptide (PBAN), having 33 amino acid residues. Antisera against syntheticHeliothis zea-PBAN were developed. Using these polyclonals, immunoreactivity was mapped in the nervous system ofAchaea janata. Three distinct groups of immunopositive secretory neurons were identified in the suboesophageal ganglion; and immunoreactivity was observed in the corpora cardiaca, thoracic and in the abdominal ganglia. From about 6000 brain sub-oesophageal ganglion complexes, the neuropeptide was isolated; and purified sequentially by Sep-pak and reversed phase high performance liquid chromatographic methods. Identity of purified PBAN fraction was confirmed with polyclonal antibody by immunoblotting. Molecular mass of the isolated peptide was determined by matrix-assisted laser desorption/ionization mass spectrometry, and was found to be 3900 Da, same as that of knownH. zea-PBAN. Radiochemical bioassay confirmed the pheromonotropic effect of the isolated neuropeptide in this insect     

Sex pheromone of the red clover casebearer moth,Coleophora deauratella,an invasive pest of clover in Canada

The red clover casebearer, Coleophora deauratella Lienig & Zeller (Lepidoptera: Coleophoridae), is an invasive pest of Trifolium species (Fabaceae) in Canada. We identified candidate sex pheromone components from female pheromone gland extracts using coupled gas chromatographic–electroantennographic analysis detection. Three compounds elicited an electrophysiological response from antennae and were identified as: (Z)‐7‐dodecenyl acetate, (Z)‐5‐dodecenyl acetate, and (Z)‐7‐dodecen‐1‐ol. Field tests of the candidate pheromone components revealed that males were attracted to a binary mixture of (Z)‐7‐dodecenyl acetate and (Z)‐5‐dodecenyl acetate. Male moth trap capture was greatest in traps baited with lures containing 100:10 or 100:20 ratios of these pheromone components, respectively. Trap capture was reduced when (Z)‐5‐dodecenyl acetate was present below 10 or above 20% of (Z)‐7‐dodecenyl acetate. Equal numbers of male moths were captured in traps baited with 10, 100, and 1 000 μg of the attractive binary mixture. These findings allow for the development of a pheromone‐based monitoring system for this invasive pest of clover in Canada.     

Female sex pheromone of Athetis lepigone (Lepidoptera: Noctuidae): Identification and field evaluation

Athetis lepigone has been recorded in many countries in Europe and Asia, but it had never been documented as an agricultural pest until 2005. For the purpose of using the sex pheromone to control this pest, we conducted a study to identify the sex pheromone of A. lepigone by gas chromatography with an electroantennographic detector (GC‐EAD) and GC coupled with mass spectrometry (GC/MS) analyses. Three pheromone candidates were detected by GC‐EAD analysis in the extracts of the female sex pheromone gland, and two candidates were identified as (Z)‐7‐dodecenyl acetate (Z7‐12:OAc) and (Z)‐9‐tetradecenyl acetate (Z9‐14:OAc) in a ratio of 1:5 by mass spectral analysis of natural pheromone components and dimethyl disulphide adducts. In the field male trapping test, the traps baited with the binary blend captured high number of males, while traps with single component hardly caught males, indicating that the two components are essential for the male attractiveness. In addition, the optimum ratios of Z7‐12:OAc and Z9‐14:OAc were determined as 3:7–7:3, and the best doses for the binary blend (at ratio of 3:7 between Z7‐12:OAc and Z9‐14:OAc) were 0.25–0.5 mg/trap, based on the number of male catches. The identification of a highly attractive sex pheromone will help in developing efficient strategies for monitoring and control of A. lepigone.     

Isolation and Structure of the Sex Pheromone Inhibitor,iAM373, of Enterococcus faecalis

An Enterococcus faecalis plasmid, pAM373, has a high frequency of transfer in a liquid medium when induced by a recipient-produced sex pheromone, cAM373. The sex pheromone inhibitor against cAM373, termed iAM373, was isolated from a culture supernatant of E. faecalis harboring pAM377 (=pAM373::Tn917), and its structure was identified as a heptapeptide, H-Ser-Ile-Phe-Thr-LeuVal-Ala-OH.     

Synthesis of (3E, 5Z)-3,5-Dodecadienylacetate,the Sex Pheromone of Phtheochroa cranaodes (Lepidoptera: Tortricidae)

(3E, 5Z)-3,5-Dodecadienyl acetate, the female sex pheromone of Phtheochroa cranaodes, was regio and stereo-selectively synthesized from 1-octyne and (E)-4-bromo-3-buten-1-ol by using Pd(PPh₃)₄, CuI and piperidine to afford the enyne (5). Further elaboration afforded the target pheromone. The synthetic pheromone was identified with the natural product by its MS and IR, data GLC retention time and biological activity.     

Semiochemistry of aposematic seed bugs

(E)-2,7-Octadienyl acetate and (E)-2-octenyl acetate (1:10 by volume) were identified as a pheromone attractive to both sexes of the lygaeid bug, Tropidothorax cruciger. In a parallel investigation of Neacoryphus bicrucis (Lygaeidae), (E,E)-2,4-hexadienyl acetate and phenethyl acetate (≈9:1) were identified from males, and found attractive to both sexes of adults in the field plus a tachinid fly parasitoid of the bugs. In N. bicrucis, the pheromone was clearly shown to come from the tubular accessory glands of the metathoracic scent gland; this evidence, plus earlier literature reports for other species, indicate that male lygaeids are the pheromone emitters. In another lygaeid, Oncopeltus fasciatus, 2-isobutyl-3-methoxypyrazine was identified in the cardiac glycoside-laden fluid sequestered from milkweed hosts and expelled by these bugs when they are attacked. Alkyl methoxypyrazines are warning odorants associated with poisonous insect secretions, and their presence in O. fasciatus indicates that the plant-derived chemical defense of lygaeines is more elaborate than previously appreciated.     

Sex pheromone of a North American population of the spotted tentiform leafminer, Phyllonorycter blancardella

The major sex pheromone compound of the spotted tentiform leafminer, Phyllonorycter blancardella (F.) (Lepidoptera: Gracillariidae), from Ontario, Canada, was identified as (10E)‐dodecen‐1‐yl acetate (E10‐12:Ac) using chemical analysis and field trapping experiments. The minor compounds (10E)‐dodecen‐1‐ol (E10‐12:OH) (4.6%), dodecan‐1‐ol (12:OH) (2.3%), and (10Z)‐dodecen‐1‐yl acetate (Z10‐12:Ac) 1.6% were also identified. The dienic acetate (4E,10E)‐dodecadien‐1‐yl acetate (E4,E10‐12:Ac), a compound reported to be attractive to P. blancardella, was not found in the glands of this population. A two‐component blend of the major and one of each the three minor compounds, in ratios similar to those found in the sex pheromone gland, did not increase the attractiveness of traps baited with synthetic pheromone. The minor compounds E10‐12:OH and 12:OH were not attractive to P. blancardella when tested individually. Z10‐12:Ac was attractive to P. blancardella, although traps baited with this compound captured only 2% of the moths that were captured in traps baited with the main compound. A four‐component blend of the major and each of the three minor compounds (100 : 1 : 1 : 1) was not more attractive than the major compound alone. The related species Phyllonorycter mespilella was captured in traps baited with E10‐12:Ac.     

Sex pheromone of the iris gelechiid moth Monochroa divisella

The iris gelechiid moth, Monochroa divisella (Douglas) (Lepidoptera: Gelechiidae), is a serious pest in iris horticulture. The overuse of pesticides for conventional control has a great risk of water pollution because irises are often planted in damp areas. To minimize pesticide applications, it is essential to monitor moth emergence by using pheromone traps. The female pheromone of M. divisella was analyzed by means of gas chromatography‐electroantennographic detection (GC‐EAD) and GC‐mass spectrometry. Two male EAD‐active compounds were detected and identified as (Z)‐3‐dodecenyl acetate (Z3‐12:Ac) and (Z)‐5‐tetradecenyl acetate (Z5‐14:Ac), the amounts of which were ca. 40 and 6 ng per female (100:15), respectively. Synthetic Z3‐12:Ac alone attracted only a few males in field trap experiments, but the attractiveness was significantly enhanced when 15% (wt/wt) Z5‐14:Ac was added; traps baited with their binary mixtures captured 21.3‐106.4 males/trap/day, as compared with 2.3 males/trap/day attracted by a crude extract of 10 virgin females. We conclude that Z3‐12:Ac and Z5‐14:Ac are M. divisella pheromone components, which can be offered as an attractant in monitoring traps. Although dodecenyl acetates are a major class of lepidopteran pheromone components, those with the double bond at the 3 position are rare and appear to be unique to gelechiids.     

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.     

Synthesis and Characterization of Hexadecadienyl Compounds with a Conjugated Diene System,Sex Pheromone of the Persimmon Fruit Moth and Related Compounds

Hexadecadien-1-ol and the derivatives (acetate and aldehyde) with a conjugated diene system have recently been identified from a pheromone gland extract of the persimmon fruit moth (Stathmopoda masinissa), a pest insect of persimmon fruits distributed in East Asia. The alcohol and acetate showed their base peaks at m/z 79 in a GC-MS analysis by electron impact ionization, but the aldehyde produced a unique base peak at m/z 84, suggesting a 4,6-diene structure. To confirm this inference, four geometrical isomers of each 4,6-hexadecadienyl compound were synthesized by two different routes in which one of two double bonds was furnished in a highly stereoselective manner. Separation of the two isomers synthesized together by each route was facilely accomplished by preparative HPLC. Their mass spectra coincided well with those of natural components, indicating that they were available for use as authentic standards for determining the configuration of the natural pheromone. Furthermore, other hexadecadienyl compounds, including the conjugated diene system between the 3- and 10-positions, were synthesized to accumulate the spectral data of pheromone candidates. 5,7-Hexadecadienal interestingly showed the base peak at m/z 80; meanwhile, the base peaks of its alcohol and acetate were detected at m/z 79 like the corresponding 4,6-dienes. The base peaks of all 6,8-, 7,9-, and 8,10-dienes universally appeared at m/z 67 like 9,11-, 10,12-, and 13,15-dienes, the spectra of which have already been published. Although 3,5-hexadecadienal was not prepared, base peaks at m/z 67 and 79 were recorded for the alcohol and acetate, respectively.     

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     

Optimization of pheromone lure and trap design for monitoring the fir coneworm, Dioryctria abietivorella

The major components of the sex pheromone of Dioryctria abietivorella (Groté) (Lepidoptera: Pyralidae) were recently identified as (9Z,11E)‐tetradecadien‐1‐yl acetate (9Z,11E‐14:Ac) and a polyunsaturated, long‐chain hydrocarbon (3Z,6Z,9Z,12Z,15Z)‐pentacosapentaene (C25 pentaene). The optimal ratio of these components and the role of potential minor components were not fully determined in the initial short report on the pheromone's identification. We tested different ratios of the two major components loaded into grey halobutyl rubber septum dispensers, placed in sticky traps deployed in conifer breeding arboreta. The optimal ratio of the two components was 200 µg 9Z,11E‐14:Ac to 2000 µg C25 pentaene. (Z)‐9‐Tetradecen‐1‐yl acetate, which had been identified previously in female pheromone gland extracts, and five other potential minor pheromone components, were tested individually as additions to the optimized two‐component lure blend. None of the ternary blends were more attractive than the optimized two‐component blend, at the ratios tested. Two lure adjuvants, a UV stabilizer (Sumisorb 300) and the antioxidant butylated hydroxytoluene, added individually or together, did not affect the attractiveness of the optimized lure blend. The Pherotech diamond sticky trap baited with the optimized lure blend was the most effective trap design among eight types of sticky trap and a bucket style trap that were tested. Traps baited with synthetic lures were as attractive as traps baited with virgin female moths. The optimized two‐component lure blend in the Pherotech diamond trap is recommended for monitoring fir coneworm infestations. The availability of an effective synthetic pheromone opens the possibility for control tactics using mating disruption or attract‐and‐kill techniques.     

Sex pheromone production and response in Korean populations of the diamondback moth, Plutella xylostella

The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Yponomeutidae), is a worldwide pest of cruciferous crops. We examined the female pheromone production and male response to various pheromone blends in two Korean populations. Gas chromatography (GC) and GC‐mass spectrometry (MS) analyses of pheromone gland extracts revealed that females produce (Z)‐11‐hexadecenal (Z11‐16:Ald), (Z)‐11‐hexadecenyl acetate (Z11‐16:OAc), and (Z)‐11‐hexadecen‐1‐ol (Z11‐16:OH) in a ratio of 8:100:18. However, (Z)‐9‐tetradecenyl acetate (Z9‐14:OAc), a previously reported component of the sex attractant of a Canadian P. xylostella population was not detected in gland extracts of the Korean one. Field tests showed that Z11‐16:Ald and Z11‐16:OAc are essential for attraction of male moths, and the highest attraction is obtained with a 10:90 blend mimicking the blend found in gland extracts. Addition of 1 or 10% of Z11‐16:OH to the 10:90 blend of Z11‐16:Ald and Z11‐16:OAc significantly increased attraction. However, attraction was strongly antagonized by the addition of as little as 0.1% of Z9‐14:OAc to the most attractive ternary blend. The ternary blend of Z11‐16:Ald, Z11‐16:OAc, and Z11‐16:OH at a ratio of 10:90:1 was more effective at catching P. xylostella males than the Japanese three‐component blend or the Canadian four‐component blend in Korea. These results suggest that there is geographical variation in the pheromone systems of this species.     

Mass‐trapping trials for the control of pine processionary moth in a pine woodland recreational area

The pine processionary moth, Thaumetopoea pityocampa, causes serious defoliation to Cedrus, Pinus and Pseudotsuga trees, as well as health problems in humans, pets and farm animals due to their urticating hairs. Environmentally friendly strategies for the management of T. pityocampa include: removal of egg batches, removal of nests, trapping of migrant larvae, spraying microbial or Insect Growth Regulator (IGR) insecticides and biocontrol, as well as pheromone‐based adult trapping and mating‐disruption. In the present paper, results on innovative technology for the control of T. pityocampa infestation using pheromone mass‐trapping are reported. Two 1‐ha plots were identified in the study area (central‐south Italy), a pine woodland recreational site growing Pinus halepensis. In the experimental plot (MT‐plot), 10 G‐traps (funnel trap type) baited with (Z)‐13‐hexadecen‐11‐ynyl acetate sex pheromone component were placed for mass‐trapping of adults; the other plot was used as a control‐plot (C‐plot). The T. pityocampa population was monitored using the two central traps in the MT‐plot and two traps positioned in the C‐plot. In addition, the winter nests made by T. pityocampa larvae overwintering on pine trees were counted. After 2 years of mass‐trapping, the number of adults trapped by the monitoring pheromone traps decreased in the MT‐plot, but not in the C‐plot, whereas the number of nests decreased in both plots. Statistical results highlighted significant differences in trap catches between the two plots but not between years. In the case of nests, differences among plots were not significant before the mass‐trapping, but significant after 1‐year treatment. According to our results, the mass‐trapping technique is able to reduce T. pityocampa infestations. This pheromone method can be applied in combination with other control systems in the context of integrated pest management in recreational areas.     

Enantiomers of fuscumol acetate comprise the aggregation‐sex pheromone of the South American cerambycid beetle Psapharochrus maculatissimus,and likely pheromones of the cerambycids Eupromerella plaumanni and Hylettus seniculus

There is increasing evidence that pheromone chemistry within the large coleopteran family Cerambycidae is often highly conserved, with numerous related species sharing the same pheromone components. As a result, traps containing these components can attract multiple cerambycid species simultaneously. In the present study, we exploited this concept in the identification of the male‐produced aggregation‐sex pheromone of the South American species Psapharochrus maculatissimus (Bates) (Coleoptera: Cerambycidae, subfamily Lamiinae, tribe Acanthoderini). Initially, live adults of both sexes were caught using a trap baited with a lure containing a blend of known cerambycid pheromone components. Headspace volatiles were collected from live beetles and analyzed by coupled gas chromatography‐mass spectrometry. Males of P. maculatissimus sex‐specifically produced a 1:38 blend of (R)‐fuscumol acetate ([2R,5E]‐6,10‐dimethylundeca‐5,9‐dien‐2‐yl acetate) and (S)‐fuscumol acetate, which were both components of the pheromone lures to which they had been attracted. In more focused field trials, traps baited with the (S)‐enantiomer, or a blend approximating the natural 1:38 ratio of (R)‐ to (S)‐enantiomers, attracted adults of both sexes in approximately equal numbers. During bioassays, adults of the lamiine species Eupromerella plaumanni (Fuchs) (tribe Acanthoderini) and Hylettus seniculus (Germar) (Acanthocinini) also were attracted, but to different lures, with E. plaumanni being attracted to the racemic mixture of the two enantiomers of fuscumol acetate, whereas H. seniculus was attracted specifically to (R)‐fuscumol acetate. Our results suggest that differences between these sympatric species in the stereochemistry of fuscumol acetate impart species‐specificity to pheromone communication channels, similar to what has been found recently with lamiine species from other continents.     

Sex pheromone components of female Cornutiplusia circumflexa

Capillary gas chromatography indicated the presence of (Z)-7-dodecenyl alcohol and (Z)-7-dodecenyl acetate in the abdominal tip extracts of female Cornutiplusia circumflexa (L.) (Lepidoptera: Noctuidae: Plusiinae) moths. Gas chromatography combined with mass spectrometry and dimethyl disulfide derivatization confirmed the structure of these two principal pheromone components. In a flight-tunnel bioassay, a 5:1 blend of alcohol and acetate elicited the complete courtship sequence by males. The response was comparable to that evoked by a live virgin female, but males spent significantly longer time at calling females than at the synthetic source. The 5:1 blend was also attractive to male C. circumflexa in the field, as indicated by catches in traps baited with this mixture. (Z)-7-dodecenyl alcohol alone was slightly attractive whereas (Z)-7-dodecenyl acetate alone was completely inactive. C. circumflexa is the first reported Plusiinae species which utilizes (Z)-7-dodecenyl alcohol as the major pheromone component.     

Attractiveness of year-old polyethylene Isonet sex pheromone dispensers for Lobesia botrana

In this study, we have compared the release of sex pheromone from mating disruption dispensers exposed in the field for 12 months and from calling females. The main pheromone component of the grapevine moth, Lobesia botrana (D. and S.) (Lepidoptera: Tortricidae), is (E)‐7,(Z)‐9‐dodecadienyl acetate, and a minor component is (Z)‐9‐dodecenyl acetate. Aged dispensers from two different years emitted a much higher amount of both pheromone components than calling females. However, the summer temperature during field exposure influenced the release from mating disruption dispensers the following year. In the wind tunnel, male L. botrana were equally attracted to 12‐month, field‐exposed dispensers, a standard monitoring pheromone lure, and to synthetic (E)‐7,(Z)‐9‐dodecadienyl acetate sprayed at the rate of 0.6–60 ng h^?1. Field trapping tests confirmed that aged dispensers from both years were at least as attractive to L. botrana males as a standard monitoring pheromone lure. The possible contribution of previously applied dispensers to the mating disruption efficacy during following applications is discussed.