Moth Pheromone Research Articles

Effects of Contact and Ingestion Exposure to Formulated CheckMate® LBAM-F and Unformulated LBAM Mating Pheromones on Adult Worker Honeybees, Apis mellifera (Hymenoptera: Apidae)

ABSTRACT Acute 7-day toxicity tests evaluating adverse effects from contact and ingestion exposure to light brown apple moth (LBAM) pheromones and time-released microencapsulated LBAM pheromones in CheckMate® LBAM-F (Checkmate) were conducted on newly emerged honeybees (less than 24 h old). Contact studies exposed bees to 1× and 10× the CheckMate label application rate. Ingestion studies exposed bees to CheckMate formulations, and active (pheromone) ingredients (a.i.) at 0.1%, 1.0%, and 10% concentrations by weight in solid food. Bees ingested approximately 39% of their body weight during the tests. Mortality ranged from 2–10% in three of four contact and ingestion exposure trials. Trial 1, which utilized a different feeding design, showed higher mortality in both control and test replicates (9–28%). One-way ANOVA tests indicated no significant difference in mortality between control and treatment replicates in the four trials. Bees were subjected to one-time CheckMate contact exposures of up to 0.49 mg/kg-bee, and average pheromone and formulation ingestion exposures of up to 56 (0.1%), 611 (1.0%), and 6,282 (10%) mg/kg-bee-day. LBAM pheromones and microencapsulated pheromones proved to be non-toxic to honeybees when sprayed with 10× the field application rate, or when ingested in food at concentrations of up to 10% by weight.

Terminal fatty-acyl-CoA desaturase involved in sex pheromone biosynthesis in the winter moth ( Operophtera brumata)

The winter moth ( Operophtera brumata L., Lepidoptera: Geometridae) utilizes a single hydrocarbon, 1,Z3,Z6,Z9-nonadecatetraene, as its sex pheromone. We tested the hypothesis that a fatty acid precursor, Z11,Z14,Z17,19-nonadecanoic acid, is biosynthesized from α-linolenic acid, through chain elongation by one 2-carbon unit, and subsequent methyl-terminus desaturation. Our results show that labeled α-linolenic acid is indeed incorporated into the pheromone component in vivo. A fatty-acyl-CoA desaturase gene that we found to be expressed in the abdominal epidermal tissue, the presumed site of biosynthesis for type II pheromones, was characterized and expressed heterologously in a yeast system. The transgenic yeast expressing this insect derived gene could convert Z11,Z14,Z17-eicosatrienoic acid into Z11,Z14,Z17,19-eicosatetraenoic acid. These results provide evidence that a terminal desaturation step is involved in the winter moth pheromone biosynthesis, prior to the decarboxylation.

Mobile mating disruption of light‐brown apple moths using pheromone‐treated sterile Mediterranean fruit flies

Public opposition to aerial application of sex pheromone for mating disruption of light-brown apple moth (LBAM), Epiphyas postvittana (Walk.), in California stopped its further use in the ca $74 million eradication programme in 2008, underscoring the need for other eradication tactics. It is demonstrated that pheromone-treated sterile Mediterranean fruit flies (medflies), Ceratitis capitata Wied., can disrupt communication in male moths. Medflies topically dosed with moth pheromone (E)-11-tetradecenyl acetate showed a no observed effect level (NOEL) of ~10 µg fly(-1) , with increasing toxicity from 30 to 100 µg fly(-1) . Greater potency and longevity of attraction and lower mortality were achieved using microencapsulated pheromone. Releases of 1000 pheromone-treated medflies ha(-1) prevented male moth catch to synthetic lures in treated 4 ha plots for 1 day in suburban Perth, Australia. Releases of ca 3000 pheromone-treated medflies ha(-1) disrupted catch to single female moths in delta traps, and to synthetic pheromone lures. Percentage disruption on the first four nights was 95, 91, 82 and 85%. Disruption of moth catch using pheromone-treated medflies is a novel development that, with future improvement, might provide a socially acceptable approach for application of the insect mating disruption technique to control invasive insects in urban environments. Adequacy of payload and other issues require resolution.

Biocontrol of moth pests in apple orchards: Preliminary field study of application potential for mass trapping

Many moth pheromones and synthetic attractants have been used to monitor or decrease pest populations. However, due to their low economic efficiencies and narrow target specificities these methods are of limited use for trapping pests in large agricultural fields. In an effort to address this problem, we selected oak (Quercus serrata) sawdust media fermented by the yeast strain Pichia anomala, and examined its ability to attract lepidopteran moth pests for mass trapping in an apple (Malus domestica) orchard. A total of 57 taxa were trapped, including 42 species of lepidopteran pests. The most frequently caught lepidopteran pests included Adoxophyes orana (40.0 ± 3.5 individuals/trap/week), Oraesia excavata (35.5 ± 2.5), and Adris tyrannus amurensis (35.5 ± 2.5). Notably, less fruit damage was recorded in the experimental orchard versus a trap-free neighboring control orchard. These results suggest that adult moths might be effectively attracted and mass captured using P. anomala-fermented oak sawdust media as bait.

Functional flexibility as a prelude to signal diversity? Role of a fatty acyl reductase in moth pheromone evolution

Sex pheromones are the hallmark of reproductive behavior in moths. Mature females perform the task of mate signaling and release bouquets of odors that attract conspecific males at long range. The pheromone chemistry follows a relatively minimal design but still the combinatorial action of a handful of specialized pheromone production enzymes has resulted in remarkably diverse sexual signals that subtly vary in structure and in number and ratio of components. In a recent article,1 we showed that a single reductase gene (pgFAR) enables the conversion of key biosynthetic fatty-acyl precursors into fatty alcohols, the immediate precursors of the multi-component pheromone in small ermine moths (Lepidoptera: Yponomeutidae). In the light of the widespread usage of multi-component pheromone blends across Lepidoptera, it is likely that the pgFAR biochemical flexibility is a regular feature of the moth pheromone machinery and polyvalent reductase genes are emerging as pivots to promote phenotypic transitions in moth mating signals. In addition, the small ermine moth pgFAR nevertheless contributes to regulating the ratio among components. Here we show that the pgFAR substrate specificity is actually counterbalancing the inherent chain-length preference of an upstream desaturase with ∆11-activity and that the enzymes together modulate the final blend ratio between the Z11-16:OH, Z11-14:OH and E11-14:OH compounds before the final acetylation.

Neofunctionalization in an ancestral insect desaturase lineage led to rare Δ 6 pheromone signals in the Chinese tussah silkworm

The Chinese tussah silkworm, Antheraea pernyi (Lepidoptera: Saturniidae) produces a rare dienoic sex pheromone composed of ( E,Z)-6,11-hexadecadienal, ( E, Z)-6,11-hexadecadienyl acetate and ( E,Z)-4,9-tetradecadienyl acetate, and for which the biosynthetic routes are yet unresolved. By means of gland composition analyses and in vivo labeling we evidenced that pheromone biosynthesis towards the immediate dienoic gland precursor, the ( E,Z)-6,11-hexadecadienoic acid, involves desaturation steps with Δ 6 and Δ 11 regioselectivity. cDNA cloning of pheromone gland desaturases and heterologous expression in yeast demonstrated that the 6,11-dienoic pheromone is generated from two biosynthetic routes implicating a Δ 6 and Δ 11 desaturase duo albeit with an inverted reaction order. The two desaturases first catalyze the formation of the ( E)-6-hexadecenoic acid or ( Z)-11-hexadecenoic acid, key mono-unsaturated biosynthetic intermediates. Subsequently, each enzyme is able to produce the ( E,Z)-6,11-hexadecadienoic acid by accommodating its non-respective mono-unsaturated product. Besides elucidating an unusually flexible pheromone biosynthetic pathway, our data provide the first identification of a biosynthetic Δ 6 desaturase involved in insect mate communication. The occurrence of this novel Δ 6 desaturase function is consistent with an evolutionary scenario involving neo-functionalization of an ancestral desaturase belonging to a gene lineage different from the Δ 11 desaturases commonly involved in moth pheromone biosynthesis.

Elucidation of the sex-pheromone biosynthesis producing 5,7-dodecadienes in Dendrolimus punctatus (Lepidoptera: Lasiocampidae) reveals Δ11- and Δ9-desaturases with unusual catalytic properties

Sex pheromones produced by female moths of the Lasiocampidae family include conjugated 5,7-dodecadiene components with various oxygenated terminal groups. Here we describe the molecular cloning, heterologous expression and functional characterization of desaturases associated with the biosynthesis of these unusual chemicals. By homology-based PCR screening we characterized five cDNAs from the female moth pheromone gland that were related to other moth desaturases, and investigated their role in the production of the ( Z)-5-dodecenol and ( Z5, E7)-dodecadienol, major pheromone constituents of the pine caterpillar moth, Dendrolimus punctatus. Functional expression of two desaturase cDNAs belonging to the Δ11-subfamily, Dpu-Δ11 1-APSQ and Dpu-Δ11 2-LPAE, showed that they catalysed the formation of unsaturated fatty acyls (UFAs) that can be chain-shortened by β-oxidation and subsequently reduced to the alcohol components. A first ( Z)-11-desaturation step is performed by Dpu-Δ11 2-LPAE on stearic acid that leads to ( Z)-11-octadecenoic acyl, which is subsequently chain shortened to the ( Z)-5-dodecenoic acyl precursor. The Dpu-Δ11 1-APSQ desaturase had the unusual property of producing Δ8 mono-UFA of various chain lengths, but not when transformed yeast were grown in presence of ( Z)-9-hexadecenoic acyl, in which case the biosynthetic intermediate ( Z9, E11)-hexadecadienoic UFA was produced. In addition to a typical Z9 activity, a third transcript, Dpu-Δ9-KPSE produced E9 mono-UFAs of various chain lengths. When provided with the ( Z)-7-tetradecenoic acyl, it formed the ( Z7, E9)-tetradecadienoic UFA, another biosynthetic intermediate that can be chain-shortened to ( Z5, E7)-dodecadienoic acyl. Both Dpu-Δ11 1-APSQ and Dpu-Δ9-KPSE thus exhibited desaturase activities consistent with the biosynthesis of the dienoic precursor. The combined action of three desaturases in generating a dienoic sex-pheromone component emphasizes the diversity and complexity of chemical reactions that can be catalysed by pheromone biosynthetic fatty-acyl-CoA desaturases in moths.

Biosynthesis of unusual moth pheromone components involves two different pathways in the navel orangeworm, Amyelois transitella.

The sex pheromone of the navel orangeworm, Amyelois transitella (Walker) (Lepidoptera: Pyralidae), consists of two different types of components, one type including (11Z,13Z)-11,13-hexadecadienal (11Z,13Z-16:Ald) with a terminal functional group containing oxygen, similar to the majority of moth pheromones reported, and another type including the unusual long-chain pentaenes, (3Z,6Z,9Z,12Z,15Z)-3,6,9,12,15-tricosapentaene (3Z,6Z,9Z,12Z,15Z-23:H) and (3Z,6Z,9Z,12Z,15Z)- 3,6,9,12,15-pentacosapentaene (3Z,6Z,9Z,12Z,15Z-25:H). After decapitation of females, the titer of 11Z,13Z-16:Ald in the pheromone gland decreased significantly, whereas the titer of the pentaenes remained unchanged. Injection of a pheromone biosynthesis activating peptide (PBAN) into the abdomens of decapitated females restored the titer of 11Z,13Z-16:Ald and even increased it above that in intact females, whereas the titer of the pentaenes in the pheromone gland was not affected by PBAN injection. In addition to common fatty acids, two likely precursors of 11Z,13Z-16:Ald, i.e., (Z)-11-hexadecenoic and (11Z,13Z)-11,13-hexadecadienoic acid, as well as traces of (Z)-6-hexadecenoic acid, were found in gland extracts. In addition, pheromone gland lipids contained (5Z,8Z,11Z,14Z,17Z)-5,8,11,14,17-icosapentaenoic acid, which also was found in extracts of the rest of the abdomen. Deuterium-labeled fatty acids, (16,16,16-D3)-hexadecanoic acid and (Z)-[13,13,14,14,15,15,16,16,16-D9]-11-hexadecenoic acid, were incorporated into 11Z,13Z-16:Ald after topical application to the sex pheromone gland coupled with abdominal injection of PBAN. Deuterium label was incorporated into the C23 and C25 pentaenes after injection of (9Z,12Z,15Z)- [17,17,18,18,18-D5]-9,12,15-octadecatrienoic acid into 1–2 d old female pupae. These labeling results, in conjunction with the composition of fatty acid intermediates found in pheromone gland extracts, support different pathways leading to the two pheromone components. 11Z,13Z-16:Ald is probably produced in the pheromone gland by Δ11 desaturation of palmitic acid to 11Z-16:Acid followed by a second desaturation to form 11Z,13Z-16:Acid and subsequent reduction and oxidation. The production of 3Z,6Z,9Z,12Z,15Z-23:H and 3Z,6Z,9Z,12Z,15Z-25:H may take place outside the pheromone gland, and appears to start from linolenic acid, which is elongated and desaturated to form (5Z,8Z,11Z,14Z,17Z)-5,8,11,14,17-icosapentaenoic acid, followed by two or three further elongation steps and finally reductive decarboxylation.

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.

Innate recognition of pheromone and food odors in moths: a common mechanism in the antennal lobe?

The survival of an animal often depends on an innate response to a particular sensory stimulus. For an adult male moth, two categories of odors are innately attractive: pheromone released by conspecific females, and the floral scents of certain, often co-evolved, plants. These odors consist of multiple volatiles in characteristic mixtures. Here, we review evidence that both categories of odors are processed as sensory objects, and we suggest a mechanism in the primary olfactory center, the antennal lobe (AL), that encodes the configuration of these mixtures and may underlie recognition of innately attractive odors. In the pheromone system, mixtures of two or three volatiles elicit upwind flight. Peripheral changes are associated with behavioral changes in speciation, and suggest the existence of a pattern recognition mechanism for pheromone mixtures in the AL. Moths are similarly innately attracted to certain floral scents. Though floral scents consist of multiple volatiles that activate a broad array of receptor neurons, only a smaller subset, numerically comparable to pheromone mixtures, is necessary and sufficient to elicit behavior. Both pheromone and floral scent mixtures that produce attraction to the odor source elicit synchronous action potentials in particular populations of output (projection) neurons (PNs) in the AL. We propose a model in which the synchronous output of a population of PNs encodes the configuration of an innately attractive mixture, and thus comprises an innate mechanism for releasing odor-tracking behavior. The particular example of olfaction in moths may inform the general question of how sensory objects trigger innate responses.

Sex-specific odorant receptors of the tobacco hornworm Manduca sexta

As odor information plays a vital role in the life of moths, their olfactory sense has evolved into a highly specific and sensitive apparatus relevant to reproduction and survival. The key players in the detection of odorants are olfactory receptor (OR) proteins. Here we identify four OR-encoding genes differentially expressed in the antennae of males and females of the sphingid moth Manduca sexta. Two male-specific receptors (the previously reported MsexOR-1 and the newly identified MsexOR-4) show great resemblance to other male moth pheromone ORs. The putative pheromone receptors are co-expressed with the co-receptor involved in general odorant signal transduction, the DmelOr83b homolog MsexOR-2. One female-specific receptor (MsexOR-5) displays similarities to BmorOR-19, a receptor in Bombyx mori tuned to the detection of the plant odor linalool.

General principles of attraction and competitive attraction as revealed by large-cage studies of moths responding to sex pheromone

Knowledge of how insects are actually affected by sex pheromones deployed throughout a crop so as to disrupt mating has lacked a mechanistic framework sufficient for guiding optimization of this environmentally friendly pest-control tactic. Major hypotheses are competitive attraction, desensitization, and camouflage. Working with codling moths, Cydia pomonella, in field cages millions of times larger than laboratory test tubes and at substrate concentrations trillions of times less than those typical for enzymes, we nevertheless demonstrate that mating disruption sufficiently parallels enzyme (ligand) -substrate interactions so as to justify adoption of conceptual and analytical tools of biochemical kinetics. By doing so, we prove that commercial dispensers of codling moth pheromone first competitively attract and then deactivate males probably for the remainder of a night. No evidence was found for camouflage. We generated and now validate simple algebraic equations for attraction and competitive attraction that will guide optimization and broaden implementation of behavioral manipulations of pests. This analysis system also offers a unique approach to quantifying animal foraging behaviors and could find applications across the natural and social sciences.

GEOGRAPHIC AND TEMPORAL VARIATION IN MOTH CHEMICAL COMMUNICATION

In moth pheromone communication signals, both quantitative and qualitative intraspecific differences have been found across geographic regions. Such variation has generally been hypothesized to be due to selection, but evidence of genetic control of these differences is largely lacking. To explore the patterns of variation in pheromone signals, we quantified variation in the female sex pheromone blend and male responses of two closely related noctuid moth species in five different geographic regions for 2-3 consecutive years. We found significant variation in the ratios of sex pheromone blend components as well as in male response, not only between geographic regions but also within a region between consecutive years. The temporal variation was of a similar magnitude as the geographic variation. As far as we know, this is the first study reporting such temporal variation in moth chemical communication systems. The geographic variation seems to at least partly be controlled by genetic factors, and to be correlated with the quality of the local chemical environment. However, the pattern of temporal variation within populations suggests that optimization of the pheromonal signal also may be driven by within-generation physiological adjustments by the moths in response to their experience of the local chemical environment.

HR11 and HR13 Receptor-Expressing Neurons Are Housed Together in Pheromone-Responsive Sensilla Trichodea of Male Heliothis virescens

The highly specific recognition of female-released sex pheromones in insects by sensory neurons of the male antenna requires specific receptors. Recently, a small family of related candidate pheromone receptors has been identified for a few moth species. In this study, the candidate pheromone receptor HR11 from Heliothis virescens has been characterized. HR11 was found to be expressed in numerous cells located in short and long sensilla trichodea on the male antenna. The HR11 cells are stereotypically arranged in a paired pattern together with HR13 cells, which respond to the major component of the sex pheromone blend. Triple in situ hybridization approaches revealed that each pair of an HR11 cell and an HR13 cell was ensheathed by supporting cells, which express pheromone-binding proteins, thus constituting a structural unit. The paired pattern of HR11/HR13 cells is reminiscent of the pattern described for BmOR-1- and BmOR-3-expressing cells in the antenna of Bombyx mori, which respond to bombykol and bombykal, respectively. These results suggest that the ligand for HR11 may be related to the HR13 ligand and furthermore imply that an arrangement of cells expressing related receptor types in the same sensillum may be a general principle in moth pheromone detection systems.

Development of sol–gel formulations for slow release of pheromones

A new type of dispenser for slow-release of sex pheromones and other semiochemicals was developed based on sol–gel polymers that can be useful for monitoring, mass trapping, and mating disruption in integrated pest management (IPM). Sol–gel matrices exhibit glass characteristics and allow control of the degree of cross-linking during the polymerization process in order to provide an optimal release rate for a particular pheromone. The advantages of sol–gel (silica) matrixes include keeping the added molecules chemically stable and enabling the sol–gel material to be applied in any desired thickness and pheromone quantity, and thereby readily modify release rates. In addition, sol–gels are primarily silica and water that are common in the environment and therefore safe for field dispensing. We developed a method for the entrapment of pheromones in sol–gel matrices that allowed release at an almost constant rate over many days in the field. For example, 2.5 mg (E)-5-decenyl acetate pheromone of peach twig borer, Anarsia lineatella, entrapped in various sol–gel formulations released 14–45 μg/day for up to 28 days. The codling moth (Cydia pomonella) pheromone in sol–gels was used in field tests to capture more codling moth males than unbaited control traps. We describe how the method may be modified to entrap other types of pheromones by making sol–gels with different pore sizes.

Scent of a Fly

Sexual courtship is a highly ritualized behavior in many animals. Recent work in the vinegar fly, Drosophila melanogaster, has illuminated how the pheromone cis-vaccenyl acetate modulates sexual behavior in the fly. Chemosensory receptors and a sexually dimorphic circuit activated by this pheromone have been identified. This minireview highlights recent advances in the field of fly courtship.

Cyclic Nucleotide-Activated Currents in Cultured Olfactory Receptor Neurons of the HawkmothManduca sexta

Moth pheromones cause rises in intracellular Ca(2+) concentrations that activate Ca(2+)-dependent cation channels in antennal olfactory receptor neurons. In addition, mechanisms of adaptation and sensitization depend on changes in cyclic nucleotide concentrations. Here, cyclic nucleotide-activated currents in cultured olfactory receptor neurons of the moth Manduca sexta are described, which share properties with currents through vertebrate cyclic nucleotide-gated channels. The cyclic nucleotide-activated currents of M. sexta carried Ca(2+) and monovalent cations. They were directly activated by cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), modulated by Ca(2+)/calmodulin, and inhibited by lanthanum. M. sexta cyclic nucleotide-activated currents developed in an all-or-none manner, which suggests that the underlying channels are coupled and act coordinately. At least one cAMP- and two cGMP-activated nonselective cation currents could be distinguished. Compared with the cAMP-activated current, both cGMP-activated currents appeared to conduct more Ca(2+) and showed a stronger down-regulation by Ca(2+)/calmodulin-dependent negative feedback. Furthermore, both cGMP-activated currents differed in their Ca(2+)-dependent inhibition. Thus M. sexta olfactory receptor neurons, like vertebrate sensory neurons, appear to express nonselective cyclic nucleotide-activated cation channels with different subunit compositions. Besides the nonselective cyclic nucleotide-activated cation currents, olfactory receptor neurons express a cAMP-dependent current. This current resembled a protein kinase-modulated low-voltage-activated Ca(2+) current.

Odor Detection in Insects: Volatile Codes

Insect olfactory systems present models to study interactions between animal genomes and the environment. They have evolved for fast processing of specific odorant blends and for general chemical monitoring. Here, we review molecular and physiological mechanisms in the context of the ecology of chemical signals. Different classes of olfactory receptor neurons (ORNs) detect volatile chemicals with various degrees of specialization. Their sensitivities are determined by an insect-specific family of receptor genes along with other accessory proteins. Whereas moth pheromones are detected by highly specialized neurons, many insects share sensitivities to chemical signals from microbial processes and plant secondary metabolism. We promote a more integrated research approach that links molecular physiology of receptor neurons to the ecology of odorants.

Tracking the sex pheromone of codling moth against a background of host volatiles with an electronic nose

Mating disruption is now widely used in apple growing regions where control of codling moth is difficult due to insecticide resistance. This strategy comprises the slow release of pheromone from controlled delivery dispensers placed on the crop field with the aim to attract and/or confuse males impeding mating. The success of the technique strongly depends on maintaining an adequate pheromone concentration in the air within the crop. At present, monitoring of the sexual confusion efficiency is done by hand using lures and traps and carrying out a weekly evaluation of the trapped insects, which involves human resources and consumes considerable time and effort. This article describes the use of an electronic nose capable of detecting and discriminating low amounts of codling moth pheromone (codlemone) immersed in a background odour of plants and fruit volatiles. Laboratory results obtained indicate the feasibility of using a trained electronic nose for tracking in real-time aerial concentration levels of codlemone against a background of common apple host volatiles. Continuous field monitoring can potentially be achieved by combining the electronic nose with automated robotic navigation and remote transference of data.

An abundant acyl-CoA (Δ9) desaturase transcript in pheromone glands of the cabbage moth, Mamestra brassicae, encodes a catalytically inactive protein

The principal sex pheromone component produced by females of the cabbage moth, Mamestra brassicae, is derived from the monounsaturated fatty acid, Z11-16:1, whereas two additional trace components are derived from E11-16:1 and Z9-16:1. This report presents the isolation and analysis of cDNAs encoding pheromone gland-specific acyl-CoA desaturases implicated in the production of these unsaturated fatty acids (UFAs). Comparisons of the encoded amino acid sequences of four cDNA fragments isolated by degenerate PCR from cabbage moth pheromone glands established their orthology with previously characterized noctuid desaturases as follows: MbraLPAQ, belonging to the pheromone gland-specific LPAQ desaturase lineage having Δ11 regioselectivity, MbraKPSE-a and MbraKPSE-b, belonging to the pheromone gland-specific KPSE desaturase lineage having Δ9 regioselectivity and a substrate preference for palmitic acid (16:0) over oleic acid (18:0), and MbraNPVE, belonging to the NPVE desaturase lineage having Δ9 regioselectivity and a substrate preference 18:0>16:0. Full-length cDNAs corresponding to the two most abundantly expressed pheromone gland-specific desaturase transcripts, MbraLPAQ and MbraKPSE-b, were isolated and assayed for their ability to genetically complement the UFA auxotrophy of a desaturase-deficient ole1 strain of Saccharomyces cerevisiae. The MbraLPAQ desaturase restored UFA prototrophy and GC–MS analysis identified Z11-16:1 and Z11-18:1 as the predominant UFAs produced. Surprisingly, MbraKPSE-b failed to complement the ole1 mutation, although it shares >98% amino acid sequence similarity with other noctuid KPSE desaturases that do. Site-directed mutagenesis of either or both of two nonconservative amino acid substitutions restored functionality to the MbraKPSE-b protein, although GC–MS analysis revealed that neither reversion resulted in the characteristic KPSE substrate preference for 16:0.