Moth Sex Research Articles

Pollinator attraction to volatiles from virgin and pollinated host flowers in a yucca/moth obligate mutualism

The classic obligate pollination–seed consumption mutualism between yuccas and yucca moths has been thought to be mediated by chemical cues, but empirical data on pollinator attraction to host floral volatiles in this association have been lacking. Here we show that the scent from virgin flowers of the host Yucca glauca is sufficient to attract its obligate pollinator Tegeticula yuccasella in Y‐tube olfactometer tests. Interestingly, both sexes of moths were attracted to the scent stimulus. Because yucca moths mate inside host flowers, the attraction of both females and males to host floral volatiles is likely to increase encounter rates. In a second test, female moths did not discriminate between virgin and hand‐pollinated flowers, indicating no post‐pollination change in scent production by the host that would lead to a reduction in pollinator attraction and thereby limit exploitation of the available seeds in host flowers. However, other mechanisms that could stabilise the mutualism between T. yuccasella and its yucca hosts have already been documented, i.e. selective abortion of heavily infested flowers, and a female‐derived host‐marking pheromone. Headspace collection and GC–MS were used to identify the blend of floral volatiles emitted by Y. glauca, which was found to be very similar to those of two other allopatric capsular‐fruited species, Y. elata and Y. filamentosa, revealing strong conservation of this trait within Yucca section Chaenocarpa.

Screening for the Genes Involved in Bombykol Biosynthesis: Identification and Functional Characterization of Bombyx mori Acyl Carrier Protein

Species-specific sex pheromones released by female moths to attract conspecific male moths are synthesized de novo in the pheromone gland (PG) via fatty acid synthesis (FAS). Biosynthesis of moth sex pheromones is usually regulated by a neurohormone termed pheromone biosynthesis activating neuropeptide (PBAN), a 33-aa peptide that originates in the subesophageal ganglion. In the silkmoth, Bombyx mori, cytoplasmic lipid droplets (LDs), which store the sex pheromone (bombykol) precursor fatty acid, accumulate in PG cells prior to eclosion. PBAN activation of the PBAN receptor stimulates lipolysis of the stored LD triacylglycerols (TAGs) resulting in release of the bombykol precursor for final modification. While we have previously characterized a number of molecules involved in bombykol biosynthesis, little is known about the mechanisms of PBAN signaling that regulate the TAG lipolysis in PG cells. In the current study, we sought to further identify genes involved in bombykol biosynthesis as well as PBAN signaling, by using a subset of 312 expressed-sequence tag (EST) clones that are in either our B. mori PG cDNA library or the public B. mori EST databases, SilkBase and CYBERGATE, and which are preferentially expressed in the PG. Using RT-PCR expression analysis and an RNAi screening approach, we have identified another eight EST clones involved in bombykol biosynthesis. Furthermore, we have determined the functional role of a clone designated BmACP that encodes B. mori acyl carrier protein (ACP). Our results indicate that BmACP plays an essential role in the biosynthesis of the bombykol precursor fatty acid via the canonical FAS pathway during pheromonogenesis.

Binding of the General Odorant Binding Protein of Bombyx mori BmorGOBP2 to the Moth Sex Pheromone Components

Insects use olfactory cues to locate hosts and mates. Pheromones and other semiochemicals are transported in the insect antenna by odorant-binding proteins (OBPs), which ferry the signals across the sensillum lymph to the olfactory receptors (ORs). In the silkworm, Bombyx mori (L.), two OBP subfamilies, the pheromone-binding proteins (PBPs) and the general odorant-binding proteins (GOBPs), are thought to be involved in both sensing and transporting the sex pheromone, bombykol [(10E,12Z)-hexadecadien-1-ol], and host volatiles, respectively. Quantitative examination of transcript levels showed that BmorPBP1 and BmorGOBP2 are expressed specifically at very high levels in the antennae, consistent with their involvement in olfaction. A partitioning binding assay, along with other established assays, showed that both BmorPBP1 and BmorGOBP2 bind to the main sex pheromone component, bombykol. BmorPBP1 also binds equally well to the other major pheromone component, bombykal [(10E,12Z)-hexadecadienal], whereas BmorGOBP2 discriminates between the two ligands. The pheromone analogs (10E,12Z)-hexadecadienyl acetate and (10E,12Z)-octadecadien-1-ol bind to both OBPs more strongly than does bombykol, suggesting that they could act as potential blockers of the response to sex pheromone by the male. These results are supported by further comparative studies of molecular docking, crystallographic structures, and EAG recording as a measure of biological response.

Phenotypic plasticity in sexual communication signal of a noctuid moth

Variability within sex pheromone signalling systems is generally believed to be low because of strong stabilizing selection; yet the noctuid moth Heliothis subflexa (Hs) shows significant intraspecific variation. One possible explanation is that females may alter their sex pheromone blend depending on prevailing olfactory cues in the habitat, which we termed the 'experience hypothesis'. This could be adaptive if Hs females experiencing the pheromone of another species, Heliothis virescens (Hv), responded to reduce the frequency of heterospecific matings. We exposed Hs females to no pheromone, Hs pheromone or Hv pheromone in the first 3 days of their adult lives. Hs females in the latter treatment produced significantly more of the acetate Z11-16:OAc, which inhibits the attraction of Hv males. To our knowledge, this is the first study showing adaptive phenotypic plasticity in a moth sex pheromone and suggests that behavioural differentiation may precede genetic divergence in the sexual signals of moths.

Effects of SPLAT® GM sprayable pheromone formulation on gypsy moth mating success

AbstractSeveral integrated pest management programs rely on the use of mating disruption tactics to control insect pests. Some programs specifically target non‐native species, such as the gypsy moth, Lymantria dispar (L.) (Lepidoptera: Lymantriidae). We evaluated SPLAT® GM, a new sprayable formulation of the gypsy moth sex pheromone disparlure, for its ability to disrupt gypsy moth mating. The study was conducted in 2006, 2007, and 2008 in forested areas in Virginia, USA. Mating success of gypsy moth females was reduced by >99% and male moth catches in pheromone‐baited traps by >90%, in plots treated with SPLAT® GM at dosages ranging from 15 to 75 g of active ingredient (a.i.) ha−1. Dosage‐response tests conducted in 2008 indicated that SPLAT® GM applied at a dosage of 7.5 g a.i. ha−1 was as effective as a 15 g a.i. ha−1 dosage.

Allelic variation in a fatty-acyl reductase gene causes divergence in moth sex pheromones

Pheromone-based behaviours are crucial in animals from insects to mammals, and reproductive isolation is often based on pheromone differences. However, the genetic mechanisms by which pheromone signals change during the evolution of new species are largely unknown. In the sexual communication system of moths (Insecta: Lepidoptera), females emit a species-specific pheromone blend that attracts males over long distances. The European corn borer, Ostrinia nubilalis, consists of two sex pheromone races, Z and E, that use different ratios of the cis and trans isomers of acetate pheromone components. This subtle difference leads to strong reproductive isolation in the field between the two races, which could represent a first step in speciation. Female sex pheromone production and male behavioural response are under the control of different major genes, but the identity of these genes is unknown. Here we show that allelic variation in a fatty-acyl reductase gene essential for pheromone biosynthesis accounts for the phenotypic variation in female pheromone production, leading to race-specific signals. Both the cis and trans isomers of the pheromone precursors are produced by both races, but the precursors are differentially reduced to yield opposite ratios in the final pheromone blend as a result of the substrate specificity of the enzymes encoded by the Z and E alleles. This is the first functional characterization of a gene contributing to intraspecific behavioural reproductive isolation in moths, highlighting the importance of evolutionary diversification in a lepidopteran-specific family of reductases. Accumulation of substitutions in the coding region of a single biosynthetic enzyme can produce pheromone differences resulting in reproductive isolation, with speciation as a potential end result.

Synthesis and biological activity of conformationally restricted gypsy moth pheromone mimics

The design and synthesis of a series of conformationally constrained mimics of gypsy moth sex pheromone, (+)-disparlure (7 R,8 S)-2-methyl-7,8-epoxyoctadecane, are described. The core structure of the mimics is derived from 5-(2′-hydroxyethyl)cyclopent-2-en-1-ol. Substituent optimization of the analogs was accomplished through the synthesis of mini-libraries and pure individual compounds, followed by electrophysiological experiments with male gypsy moth antennae. The electroantennogram results show that the analogs elicited weak to no antennal responses themselves. There was a clear structure–activity pattern for odorant activity, with ethyl substituents being best. Further, when puffed simultaneously with the pheromone, some of the compounds gave a significant enhancement of the antennal depolarization, indicating an additive or synergistic effect. A pure pheromone stimulus following a mixed compound/pheromone stimulus was generally not affected, with two exceptions: one compound enhanced and another inhibited a subsequent stimulus. The compounds also prolonged the stimulation of the antenna, which manifested itself in widened electroantennogram peaks. We tested the hypothesis that this prolonged stimulation may be due to the stabilization of a particular conformer of the pheromone-binding protein (PBP). Compounds that caused PBP2 to adopt a similar conformation than in the presence of pheromone also caused peak widening. This was not the case with PBP1.

Selectivity of the oriental fruit moth sex pheromone trap in peach and apricot orchards

One of the most important pests of the stone fruit orchards is the oriental fruit moth (Grapholitha 1110/esta B.). Chemical control targeted against the young larvae is the most effective way of protection. so the ti ming of treatments has to be based on the observation of emergence. Emergence may be monitored with sex pheromone traps. It is already known from former publications, that the traps for oriental fruit moth are also effective in the case of the plum moth (Grapholitha f1111ebra11a Tr.), which external morphology is very similar to the oriental fruit moth. As the emergence of the oriental fruit moth in peach and apricot orchards has not been observed in detaib in Hungary, we started a s1Udy in this field. Our aim was to measure the selectivity of the sex pheromone traps. On the basis of examining more than 5000 males caught and the investigation of male genital ia. it could be established that the pheromone traps. Csalomon and Deltastop, for oriental fruit moth, caught the plum moth in the same ratio. The ratio of the oriental fruit moth and the plum moth trapped in the peach orchards was I: I . while in the apricot orchards the number of the caught plum moth males was seven times as many as that of the oriental fruit moths. Consequently, it can be established that data based on oriental fruit moth trap catches can not be used without additional investigations of genitalia for the prediction of larval hatch. The selectivity of the plum moth trap. used as a control. was acceptable in both orchards.

Attraction of DifferentTrichogrammaSpecies toPrays oleaeSex Pheromone

AbstractChemical cues play a major role in the process of host selection by parasitoids, a process that has been discriminated into several steps such as habitat location, host location, and host acceptance. Egg parasitoids are known to rely on infochemicals of the adult host stage, e.g., pheromones, rather than cues emitted by the inconspicuous host eggs themselves. Here, we show that three different Trichogramma (Hymenoptera: Trichogrammatidae) species were attracted to the synthetic sex pheromone of the olive, Olea europea L., pest Prays oleae (Bern) (Lepidoptera: Yponomeutidae). In Y-tube olfactometer experiments, we revealed differences in the behavior of four tested Trichogramma strains to different concentrations of the host pheromone. Although wasps of Trichogramma oleae (Voegele & Poitale) (IO47) were significantly attracted to two concentrations of the host pheromone, another strain of the same species (IO24) did not show a response to any offered concentrations. Trichogramma cacoeciae Marchai females showed greatest attraction toward the pheromone to three of five offered concentrations. Bioassays with Trichogramma bourarachae Pintureau & Babault wasps showed a positive response of the parasitoids to three concentrations of the host pheromone. Detection of the moth sex pheromone may result in an enhanced searching effort by Trichogramma wasps in an area with host eggs and eventually lead to an increased parasitism rate of these eggs. The results of this research should help enhancing the efficacy of the use of Trichogramma wasps as biocontrol agents against the olive pest.

Olfactory attraction of egg parasitoids to virgin females of noctuid stemborers

To locate hosts, egg parasitoids rely on infochemicals of the adult host stage, e.g. pheromones, rather than cues emitted by the inconspicuous egg themselves. Here, we show that three different egg parasitoid species the scelionids Telenomus busseolae Gahan and Telenomus isis Polaszek and the trichogrammatid Trichogramma bournieri Pintureau & Babault were attracted to both calling and non-calling females of the noctuids Busseola fusca (Fuller), Sesamia calamistis (Hampson) and Sesamia nonagrioides (Lefebvre). In Y-tube olfactometer experiments this study revealed a preference of all three parasitoids for non-calling (general odors of virgin females) and calling moth (sex pheromone) over the control (clean air), and for calling over the non-calling moth. However, the three parasitoids were equally attracted to calling moth of B. fusca and S. calamistis indicating low host specificity. The findings indicated that all three parasitoids used the pheromones released by the calling moth in host finding. It is suggested that the low host specificity may affect egg parasitism of the target pest in crop fields.

Evolution of Moth Sex Pheromone Desaturases

Moth sex pheromone communication has evolved to use complex blends of relatively simple long-chain fatty acid precursors. Species specificity is derived from the unique stereochemistry of double bonds introduced into exact locations along the hydrocarbon backbone of fatty acids, which are reduced and then undergo a variety of chain-shortening and functionalization reactions to form the pheromone blend. Key enzymes that have evolved to function in this system are the acyl coenzyme A desaturases, which catalyze the introduction of the double bonds. This report gives an overview of the evolution of these enzymes, with an introduction to the newly arisen field of "semiochemical genetics."

Nearest Neural Neighbors: Moth Sex Pheromone Receptors HR11 and HR13

In moth sex pheromone olfaction systems, there is a stereotypical co-compartmentalization of two or sometimes three olfactory receptor neurons (ORNs) within single trichoid sensilla on which pheromone-sensitive odorant receptors (ORs) are differentially expressed. In this issue of Chemical Senses, Krieger et al. show through elegant double and triple in situ hybridization studies that in the moth, Heliothis virescens, a pheromone component-related OR (HR11) is expressed on an ORN that is reliably cocompartmentalized in the same sensillum as another OR (HR13) whose ligand is known to be (Z)-11-hexadecenal, the H. virescens major pheromone component. Although the ligand for HR11 is not yet known, mapping this OR to this particular ORN represents a key advance in piecing together the puzzle of H. virescens sex pheromone olfaction.

Gypsy moth mating disruption in open landscapes

Abstract1 Aerial applications of Disrupt II, a plastic laminated flake formulation containing a racemic form of the gypsy moth sex pheromone, disparlure, achieved > 99% reduction of mating among females on individual, isolated trees surrounded by an area cleared of trees.2 These results support the use of mating disruption to eradicate isolated gypsy moth populations in open landscapes, such as parks, residential areas and commercial settings.3 Mating success in both treated and untreated areas varied with the initial distance between males and females. When the initial distance between males and females was < 5 cm in an area receiving a dosage of 37.5 g of racemic disparlure per ha, mating success was reduced by 27% compared with a similar deployment in an untreated area. Mating was eliminated in areas treated at the same dosage when males and females were initially deployed 1 m apart but on separate trees.4 This suggests that mating disruption may not be an effective tactic for gypsy moth eradication in cases where the infestation is concentrated on a small number of trees and males and females are in close proximity in space and time.

Effects of the 3M™ MEC Sprayable Pheromone® formulation on gypsy moth mating success

AbstractThe study was conducted during 2000, 2001, 2003 and 2004 in forested areas in Virginia, USA to evaluate the 3M™ MEC‐GM Sprayable Pheromone® formulation of the gypsy moth sex pheromone, disparlure, for its ability to disrupt mating in gypsy moth, Lymantria dispar (Lep.: Lymantriidae). Both mating success of gypsy moth females and male moth catches in pheromone‐baited traps were significantly reduced in plots treated with the 3M™ MEC‐GM formulation at dosages ranging from 15 to 75 g of active ingredient/ha. However, the 3M™ MEC‐GM formulation reduced trap catch to a lesser extent than did the currently registered Hercon Disrupt® II plastic flakes used as a positive control and applied at similar or lower dosages. Furthermore, the effectiveness of the 3M™ sprayable formulation declined through time, so that by the end of the male flight season, male moth catches in traps were significantly higher than in plots treated with Hercon plastic flakes. Based on the reported results, 3M™ MEC‐GM Sprayable Pheromone® formulation was never integrated into the operational treatment projects of USDA Forest Service Cooperative Slow‐the‐Spread of the Gypsy Moth management programme.

Balanced Olfactory Antagonism as a Concept for Understanding Evolutionary Shifts in Moth Sex Pheromone Blends

In the sex pheromone communication systems of moths, both heterospecific sex pheromone components and individual conspecific pheromone components may act as behavioral antagonists when they are emitted at excessive rates and ratios. In such cases, the resulting blend composition does not comprise the sex pheromone of a given species. That is, unless these compounds are emitted at optimal rates and ratios with other compounds, they act as behavioral antagonists. Thus, the array of blend compositions that are attractive to males is centered around the characterized female-produced sex pheromone blend of a species. I suggest here that the resulting optimal attraction of males to a sex pheromone is the result of olfactory antagonistic balance, compared to the would-be olfactory antagonistic imbalance imparted by behaviorally active compounds when they are emitted individually or in other off-ratio blends. Such balanced olfactory antagonism might be produced in any number of ways in olfactory pathways, one of which would be mutual, gamma-aminobutyric-acid-related disinhibition by local interneurons in neighboring glomeruli that receive excitatory inputs from pheromone-stimulated olfactory receptor neurons. Such mutual disinhibition would facilitate greater excitatory transmission to higher centers by projection interneurons arborizing in those glomeruli. I propose that in studies of moth sex pheromone olfaction, we should no longer artificially compartmentalize the olfactory effects of heterospecific behavioral antagonists into a special category distinct from olfaction involving conspecific sex pheromone components. Indeed, continuing to impose such a delineation among these compounds may retard advances in understanding how moth olfactory systems can evolve to allow males to exhibit correct behavioral responses (that is, attraction) to novel sex-pheromone-related compositions emitted by females.

Facile Synthesis of Mill Moth's Sex Pheromone Components

An improved method for the preparation of mill moth's sex pheromone components in high diastereomeric purity is described.

Structure–activity relationships of pheromonostasis induced by ACCase-inhibitor herbicides in the moth Helicoverpa armigera

Many moth sex pheromone blends are derived from fatty acids and their production is regulated by a Pheromone Biosynthesis Activating Neuropeptide (PBAN). In prior work we showed that the herbicide Diclofop-acid, an acetyl-coenzyme A carboxylase (ACCase) inhibitor, inhibits PBAN-induced sex pheromone production in vitro. In this work we extend our study showing that several other herbicides, belonging to the 2-aryloxyphenoxypropionate (‘FOP’) and the cyclohexandione-oxime (‘DIM’) families significantly inhibit pheromone production by adult females whilst survival is unaffected by treatment. Enzyme activity in vitro and kinetic analysis revealed a K m of 0.35 μM with K i values of 0.1 and 0.28 μM due to Tralkoxydim and Diclofop inhibition, respectively. Inhibitory activity on PBAN-induced pheromone production by all herbicides tested revealed a potency order: Tralkoxydim > Clodinafop > Cycloxidim > Haloxyfop > Diclofop > Fenoxaprop > Fluazifop > Quizalofop, Quizalofop being inactive. Differences in inhibition efficiencies may be attributed to different binding sites on the enzyme or to the polarity and solubility of these compounds.

Efficacy of the pear ester as a monitoring tool for codling moth Cydia pomonella (Lepidoptera: Tortricidae) in New Zealand apple orchards

The behavioural response of both sexes of codling moth, Cydia pomonella to the pear-derived kairomone (ethyl (2E,4Z)-2,4-decadienoate), codling moth sex pheromone (E,E-8,10-dodecadien-1-ol), and sex pheromone combined with the pear derived kairomone loaded into red rubber septum were investigated in trapping experiments in New Zealand apple orchards. A range of 0.01-10.0 mg of pheromone loading in rubber septum dispensers was tested and the highest catch of males was in traps baited with 1.0 mg. No dose response in trap catch of males was seen in traps baited with different amounts of pear-derived kairomone (0.01-10.0 mg). The number of females caught was significantly affected by the amount of pear derived kairomone used to bait traps, with the highest catch obtained at 10 mg loading. The attractiveness of sex pheromone was not enhanced by the addition of the kairomone either when used in the same bait or in a separate bait. The mean number of males captured in traps was reduced by 44% when the pheromone and kairomone were combined at ratio of 1:1 (0.1 mg pheromone: 0.1 mg kairomone) in separate sources. Kairomone baited traps showed some potential for monitoring the flight activity of female C. pomonella in apple orchards in two locations (Canterbury and Hawke's Bay). However, the number of male moths caught was low as compared to the number of male moths caught in pheromone-baited traps, and therefore the sex pheromone should continue to be used for monitoring male activity.

Codling Moth Management and Chemical Ecology

Lepidopteran insects use sex pheromones to communicate for mating. Olfactory communication and mate-finding can be prevented by permeating the atmosphere with synthetic pheromone. Pheromone-mediated mating disruption has become a commercially viable pest management technique and is used to control the codling moth, Cydia pomonella, a key insect pest of apple, on 160,000 ha worldwide. The codling moth sex pheromone, codlemone, is species specific and nontoxic. Orchard treatments with up to 100 grams of synthetic codlemone per hectare effectively control codling moth populations over the entire growing season. Practical implementation of the mating disruption technique has been realized at an opportune time, as codling moth has become resistant to many insecticides. We review codling moth chemical ecology and factors underlying the behavioral mechanisms and practical implementation of mating disruption. Area-wide programs are the result of collaborative efforts between academic research institutions, extension, chemical industries, and grower organizations, and they demonstrate the environmental and economic relevance of pheromone research.

Persistent effects of aerial applications of disparlure on gypsy moth: trap catch and mating success

AbstractIn forest plots treated aerially with a plastic laminated flake formulation (Disrupt® II) of the gypsy moth sex pheromone disparlure to disrupt gypsy moth, Lymantria dispar (L.) (Lepidoptera: Lymantriidae), mating was monitored the year of treatment and 1–2 years after treatment to determine the effects of the treatment on suppression of trap catch and mating success. In the year of treatment, there was a greater than 95% reduction in trap catch and a greater than 98% reduction in mating success compared to controls. One year after treatment at a dosage of 37.5 g active ingredient (a.i.) ha−1, trap catch was reduced by 46–56% and mating success was reduced by 60–79%. Both trap catch and mating success were significantly reduced compared to controls in plots treated 1 year previously at 15 g a.i. ha−1. Trap catch, but not mating success, was significantly reduced 2 years after treatment at 37.5 g a.i. ha−1. The efficacy of mating disruption (MD) treatments in the Slow‐the‐Spread of the Gypsy Moth program was significantly reduced 2 years compared to 1 year after treatment. No such reduction was observed in plots treated with aerial applications of Bacillus thuringiensis kurstaki. The higher apparent efficacy of MD treatments 1 year after application may result to some extent from the suppression of moth capture in pheromone traps from the persistent effects of the previous year's treatment.