Pheromone Races Research Articles

Plant Preferences of Z-Pheromone Race Ostrinia nubilalis (Lepidoptera: Crambidae) Based on Leaf Tissue Consumption Rates.

European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), has been present in the United States for over 100 yr and documented on >200 plant species, including economically valuable crops. The reported preferred host of O. nubilalis is corn, Zea mays L. (Cyperales: Poaceae), although it is considered to be a generalist agricultural pest. Life cycles of the two pheromone races, E and Z, align with the seasonality of different agricultural plants. Since the introduction of Bt corn in 1996, overall O. nubilalis presence has declined and suggests that alternative crop plants might not be suitable hosts. We investigated plant vegetation preference of third-instar Z-race O. nubilalis for leaf disks of corn and a variety of other crops using 48 h no-choice and choice tests. Z-race larvae gained more mass on V6 non-Bt field corn leaf disks in comparison to other plant species options. Additionally, a preference for non-Bt field corn leaf disks was observed in most comparisons. Higher consumption of cucumber, Cucumis sativus L. (Cucurbitales: Cucurbitaceae), leaf disks as compared to non-Bt field corn leaf disks suggested an ability to feed on excised leaf tissues of a plant species that does not induce defenses to herbivory.

Genetic mapping of male pheromone response in the European corn borer identifies candidate genes regulating neurogenesis

The sexual pheromone communication system of moths is a model system for studies of the evolution of reproductive isolation. Females emit a blend of volatile components that males detect at a distance. Species differences in female pheromone composition and male response directly reinforce reproductive isolation in nature, because even slight variations in the species-specific pheromone blend are usually rejected by the male. The mechanisms by which a new pheromone signal-response system could evolve are enigmatic, because any deviation from the optimally attractive blend should be selected against. Here we investigate the genetic mechanisms enabling a switch in male response. We used a quantitative trait locus-mapping approach to identify the genetic basis of male response in the two pheromone races of the European corn borer, Ostrinia nubilalis Male response to a 99:1 vs. a 3:97 ratio of the E and Z isomers of the female pheromone is governed by a single, sex-linked locus. We found that the chromosomal region most tightly linked to this locus contains genes involved in neurogenesis but, in accordance with an earlier study, does not contain the odorant receptors expressed in the male antenna that detect the pheromone. This finding implies that differences in the development of neuronal pathways conveying information from the antenna, not differences in pheromone detection by the odorant receptors, are primarily responsible for the behavioral response differences among the males in this system. Comparison with other moth species reveals a previously unexplored mechanism by which male pheromone response can change in evolution.

Pheromone races of Cydia splendana (Lepidoptera, Tortricidae) overlap in host plant association and geographic distribution

Identification of the sex pheromone of Cydia splendana (Lepidoptera, Tortricidae) by pheromone gland analysis followed by field trapping with synthetic compounds shows the occurrence of two pheromone races. Acorn moth females from Sweden, where oak Quercus robur is the only host plant, use a blend of the E,Z and E,E isomers of 8,10-dodecadien-1-yl acetate. In Central and Southern Europe, where C. splendana feeds on chestnut Castanea sativa and several species of oak, males respond to another isomer blend, E,E and Z,E. The distribution of the two pheromone races of C. splendana overlaps in Northern France, where they share oak as plant host. Differences in sex communication signals lead to behavioural pre-mating isolation between these populations, and emphasize the role of specific mate recognition in speciation events.

Frequency of hybridization between Ostrinia nubilalis E-and Z-pheromone races in regions of sympatry within the United States.

Female European corn borer, Ostrinia nubilalis, produce and males respond to sex pheromone blends with either E- or Z-Δ11-tetradecenyl acetate as the major component. E- and Z-race populations are sympatric in the Eastern United States, Southeastern Canada, and the Mediterranean region of Europe. The E- and Z-pheromone races of O. nubilalis are models for incipient species formation, but hybridization frequencies within natural populations remain obscure due to lack of a high-throughput phenotyping method. Lassance et al. previously identified a pheromone gland-expressed fatty-acyl reductase gene (pgfar) that controls the ratio of Δ11-tetradecenyl acetate stereoisomers. We identified three single nucleotide polymorphism (SNP) markers within pgfar that are differentially fixed between E- and Z-race females, and that are ≥98.2% correlated with female pheromone ratios measured by gas chromatography. Genotypic data from locations in the United States demonstrated that pgfar-z alleles were fixed within historically allopatric Z-pheromone race populations in the Midwest, and that hybrid frequency ranged from 0.00 to 0.42 within 11 sympatric sites where the two races co-occur in the Eastern United States (mean hybridization frequency or heterozygosity (HO) = 0.226 ± 0.279). Estimates of hybridization between the E- and Z-races are important for understanding the dynamics involved in maintaining race integrity, and are consistent with previous estimates of low levels of genetic divergence between E- and Z-races and the presence of weak prezygotic mating barriers.This work describes the development of new single nucleotide polymorphism (SNP) markers within the pheromone gland expressed fatty acyl reductase (pgfar) gene of Ostrinia nubilalis. These SNPs were shown to segregate based upon female pheromone production, and thus provide the first description of an assay for genetic determination of O. nubilalis pheromone strain from field-collected samples. These assays were applied to estimate hybridization within field populations, and represent valuable tools for future population genetic studies of this species.

Volatile organic compounds and host-plant specialization in European corn borer E and Z pheromone races

Plant volatile cues are considered the main source of information for ovipositing moths, which use chemical information to locate and recognize the host plant. In Europe, two sympatric populations of European corn borer (ECB; Ostrinia nubilalis, Hubner), the Z and E-pheromone races, feed mainly on maize and hop or mugwort, respectively. We studied the mechanisms of host-plant recognition and fidelity in ECB pheromone races by testing the attractiveness of host plants to gravid females in a flight tunnel and by analyzing the volatiles released from maize, mugwort, and hop during the scotophase, when the ovipositing flight of the ECB females occurs. In the wind tunnel bioassay, the Z-race and E-race females engaged in upwind flight and expressed a strong host fidelity to their respective main host plants; all three of these host plants possess distinctive volatile profiles specific as to blend and ratio. The host plants shared a certain number of ubiquitous volatiles present in various ratios that likely constitute a species-specific cue to host-seeking ECB moths. Our observations therefore suggest that ECB host fidelity is steered by plant volatiles that are present in species-specific ratios of ubiquitous volatile organic compounds.

Asian corn borer pheromone binding protein 3, a candidate for evolving specificity to the 12-tetradecenyl acetate sex pheromone

Most moth species in the genus Ostrinia use varying ratios of (Z)-11- and (E)-11-tetradecenyl acetate as their main sex pheromone components. The Asian corn borer is unique within the genus having evolved to use pheromone components with a shift in the location of the double bond, (Z)-12- and (E)-12-tetradecenyl acetate. We identified cDNAs representing five pheromone binding proteins (PBPs) and two sensory neuron membrane protein genes from an antennal transcriptome. The coding regions of the orthologous genes were cloned from the Asian corn borer and the (E) and (Z) sex pheromone races of the European corn borer. Their nucleotide sequences and transcript expression levels were analyzed to identify candidate genes from the Asian corn borer that may have evolved specificity to the 12-tetradecenyl acetate ligand. PBP2 and PBP3 transcripts were expressed at high male-biased levels. PBP3 had the most nonsynonymous nucleotide substitutions resulting in ten amino acid changes. Based on the predicted three-dimensional structure of PBP3, six of these ten amino acid changes occur in domains that may interact with the pheromone ligand. Future studies will determine whether PBP3 has evolved specificity to the Asian corn borer sex pheromone.

The Predictive Research of Lymantria Monacha Species within Natura 2000 Cusma Site

SYMMARY Lymantria monacha is one of the most dangerous butterfly species, af fecting softwood in general ( spruce, pine, fir, larch, duglas ), but also hardwood in particular. The l arvaes attack in the beginning the young buds just after their fr uit outlets, ending the attack by eating, total or partial, the needles. The species has a large pr opagation, often leeding to the drying of trees, causing up to 60% defoliation (Oltean et. al l, 2003). For the quantitative evaluation of Lymantria monacha population within Natura 2000 Cusma site, sex attractant pheromone races have been placed during 2008, within the terr itory of Tihu ţa-Colibiţa, Josenii Bârg aului and Bistri ţa Bârg aului Forest Districts (Proorocu et. all, 2007, 2009). Within Tihu ţa-Colibiţa Forest District territory, 27 pheromone races hav e been installed, distributed both in the young and in the mature bru shes. The medium number of individuals/race was 30 and the maximum number of individuals (101) has been collected by race number 4, race that has been places within a 7 5 years old forest, formed by a mix of spruce, fir and beech, with a spruce predomination. The recorded annual dynamic is characteristic for the species that develop only on e generation per year, with a numerical maximum in the beginning of August. Within Josenii Bârg aului Forest District territory, 58 pheromone races have been installed in an over 60 years old mature brush, a m edium number of 18 individuals/race and a maximum number of 43 individuals/race being recorded. Within Bistri ţa Bârg aului Forest District territory, 94 pheromone races have been in stalled in an over 60 years old mature brush, the medium number of Lymantria monacha individuals/race being 47, and the maximum one, 99. The achieved data allows us to conclude the fact that, within Natura 2000 Cusma Site, the attack of Lymantria monacha species is a moderate one, no other detection methods beeing necesary.

Managing resistance to Bt crops in a genetically variable insect herbivore, Ostrinia nubilalis

To slow the resistance evolution of the European corn borer (ECB) to Cry proteins expressed in transgenic Bacillus thuringensis (Bt) corn, the United States Environmental Protection Agency (EPA) has adopted an insect resistance management (IRM) plan that relies on a "high dose/refuge" strategy. However, this IRM plan does not consider possible ecological differences between the two ECB pheromone races (E and Z). Using carbon isotope analysis, we found that unstructured (non-corn) refuges contribute more to E race (18%) than to Z race (4%) populations of ECB in upstate New York (USA). Furthermore, feeding on non-corn hosts is associated with decreased body mass and reduced fecundity. We also show that the geographic range of E-race ECB is restricted within the range of the Z race and that E-race ECB are increasingly dominant in regions with increasing non-corn habitat. While the proportion of E-race ECB developing in unstructured refuges is higher than previously assumed, low rates of unstructured refuge use by the Z race, evidence for reduced fecundity when reared on non-corn hosts, and complete sympatry within the E race range all argue against a relaxation of current IRM refuge standards in corn based on alternative-host use. We also discuss implications of this research for integrated pest management in vegetables and IRM in Bt cotton.

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.

Sex pheromone receptor specificity in the European corn borer moth, Ostrinia nubilalis.

BackgroundThe European corn borer (ECB), Ostrinia nubilalis (Hubner), exists as two separate sex pheromone races. ECB(Z) females produce a 97∶3 blend of Z11- and E11-tetradecenyl acetate whereas ECB(E) females produce an opposite 1∶99 ratio of the Z and E isomers. Males of each race respond specifically to their conspecific female's blend. A closely related species, the Asian corn borer (ACB), O. furnacalis, uses a 3∶2 blend of Z12- and E12-tetradecenyl acetate, and is believed to have evolved from an ECB-like ancestor. To further knowledge of the molecular mechanisms of pheromone detection and its evolution among closely related species we identified and characterized sex pheromone receptors from ECB(Z).MethodologyHomology-dependent (degenerate PCR primers designed to conserved amino acid motifs) and homology-independent (pyrophosphate sequencing of antennal cDNA) approaches were used to identify candidate sex pheromone transcripts. Expression in male and female antennae was assayed by quantitative real-time PCR. Two-electrode voltage clamp electrophysiology was used to functionally characterize candidate receptors expressed in Xenopus oocytes.ConclusionWe characterized five sex pheromone receptors, OnOrs1 and 3–6. Their transcripts were 14–100 times more abundant in male compared to female antennae. OnOr6 was highly selective for Z11-tetradecenyl acetate (EC50 = 0.86±0.27 µM) and was at least three orders of magnitude less responsive to E11-tetradecenyl acetate. Surprisingly, OnOr1, 3 and 5 responded to all four pheromones tested (Z11- and E11-tetradecenyl acetate, and Z12- and E12-tetradecenyl acetate) and to Z9-tetradecenyl acetate, a behavioral antagonist. OnOr1 was selective for E12-tetradecenyl acetate based on an efficacy that was at least 5-fold greater compared to the other four components. This combination of specifically- and broadly-responsive pheromone receptors corresponds to published results of sensory neuron activity in vivo. Receptors broadly-responsive to a class of pheromone components may provide a mechanism for variation in the male moth response that enables population level shifts in pheromone blend use.

Homology of olfactory receptor neuron response characteristics inferred from hybrids between Asian and European corn borer moths (Lepidoptera: Crambidae)

First generation hybrid males from crosses between the Asian corn borer (ACB), Ostrinia furnacalis, and the “univoltine Z-strain” European corn borer (ECB), Ostrinia nubilalis, were examined with respect to behavioral and physiological responses to ACB and ECB pheromones. The hybrid males often flew to the pheromone of ECB Z-strain, but very rarely to the ACB pheromone. We mapped the tuning profiles of each ORN of the F1 hybrids with respect to the relevant pheromone components and a common behavioral antagonist by employing differential cross-adaptation and varying doses of the ligands. In the trichoid sensilla of F1 hybrid males, the three co-compartmentalized ORNs produced spikes that were very difficult to distinguish by size, unlike the parental populations. Comparing the responses to ACB and ECB components at different doses reveals overlapping profiles similar to males of both parental types, but more responsiveness to the ECB pheromone components. We were unable to detect any differences in the ORN tuning profiles when comparing males with different behavioral phenotypes. While the two ECB pheromone races have similar ORN tuning properties that are different from those in ACB, the spike-amplitude patterns of ECB E-strain and ACB have greater homology when compared to ECB Z-strain.

Pheromone identification ofDioryctria abietivorella(Lepidoptera: Pyralidae) from an eastern North American population: geographic variation in pheromone response

AbstractThe fir coneworm,Dioryctria abietivorella(Groté), is a transcontinental pest of coniferous seed cones. Recently, the attractive pheromone components and lure blend for the fir coneworm in western Canada were identified as a 200 µg : 2000 µg ratio of (9Z,11E)-tetradecadienyl acetate (9Z,11E-14:Ac) and a polyunsaturated hydrocarbon, (3Z,6Z,9Z,12Z,15Z)-pentacosapentaene (C25pentaene). A potential third component, (Z)-9-tetradecenyl acetate (9Z-14:Ac), which was detected in pheromone gland extracts, had no effect on males of the western population when added to the binary blend. In contrast, in eastern Canada (Quebec) we found that the 200 µg : 2000 µg binary blend was unattractive to male fir coneworms until 9Z-14:Ac was added. By means of several field experiments we established that a 67 µg : 200 µg : 2000 µg ratio of 9Z-14:Ac, 9Z,11E-14:Ac, and the C25pentaene was attractive to fir coneworms in Quebec. Gas chromatography - mass spectrometry analysis of pheromone gland extracts of fir coneworm from Quebec confirmed the presence of the three components in extracts in an approximately 1 µg : 4 µg : 10 µg ratio of 9Z-14:Ac, 9Z,11E-14:Ac, and the C25pentaene. Our results provide evidence that the eastern and western populations ofD. abietivorellaare distinct geographic pheromone races, with 9Z-14:Ac being required for attracting males of the eastern population but not males of the western population.

Experimental Evidence for Three Pheromone Races of the Scarab Beetle Phyllophaga anxia (LeConte)

This study offers experimental evidence for the existence of three pheromone races of the northern genitalic form of Phyllophaga anxia: one race in which females produce and males respond mainly to L-valine methyl ester, a second producing and responding to L-isoleucine methyl ester, and a third producing and responding to an intermediate range of blends of the two compounds. At Franklinville, NY, pheromone gland contents of females were analyzed using coupled gas chromatography-electroantennogram detection. Two types of females were found, one that produced greater than 99% L-valine methyl ester and another that produced greater than 99% L-isoleucine methyl ester. Capture-mark-release-recapture field tests with males at Franklinville established that most males were recaptured in traps baited with the same blends with which they were originally captured. The populations characterized at Franklinville, NY, have also been found at numerous locations from eastern Canada and the northeast and north central USA, sometimes in allopatry and sometimes in sympatry. At a site in Carver, MA, P. anxia males responded to blends of the methyl esters of L-valine and L-isoleucine, and Carver females produced blends similar to those to which the males responded. Populations responding to blends have been identified only from southeastern Massachusetts and Rhode Island. At a field site near Waterloo, NY, the addition of small proportions of L-isoleucine methyl ester to lures containing L-valine methyl ester did not affect trap captures, but higher proportions of L-isoleucine methyl ester were inhibitory, decreasing trap captures.

Monitoring of European Corn Borer with Pheromone-Baited Traps: Review of Trapping System Basics and Remaining Problems

Since the identification of female European corn borer, Ostrinia nubilalis (Hübner) pheromone, pheromone-baited traps have been regarded as a promising tool to monitor populations of this pest. This article reviews the literature produced on this topic since the 1970s. Its aim is to provide extension entomologists and other researchers with all the necessary information to establish an efficient trapping procedure for this moth. The different pheromone races of the European corn borer are described, and research results relating to the optimization of pheromone blend, pheromone bait, trap design, and trap placement are summarized followed by a state-of-the-art summary of data comparing blacklight trap and pheromone-baited trap techniques to monitor European corn borer flight. Finally, we identify the information required to definitively validate/invalidate the pheromone-baited traps as an efficient decision support tool in European corn borer control.

Monitoring of European Corn Borer with Pheromone-Baited Traps: Review of Trapping System Basics and Remaining Problems

Since the identification of female European corn borer, Ostrinia nubilalis (Hübner) pheromone, pheromone-baited traps have been regarded as a promising tool to monitor populations of this pest. This article reviews the literature produced on this topic since the 1970s. Its aim is to provide extension entomologists and other researchers with all the necessary information to establish an efficient trapping procedure for this moth. The different pheromone races of the European corn borer are described, and research results relating to the optimization of pheromone blend, pheromone bait, trap design, and trap placement are summarized followed by a state-of-the-art summary of data comparing blacklight trap and pheromone-baited trap techniques to monitor European corn borer flight. Finally, we identify the information required to definitively validate/invalidate the pheromone-baited traps as an efficient decision support tool in European corn borer control.

Assortative Mating between European Corn Borer Pheromone Races: Beyond Assortative Meeting

BackgroundSex pheromone communication systems may be a major force driving moth speciation by causing behavioral reproductive isolation via assortative meeting of conspecific individuals. The ‘E’ and ‘Z’ pheromone races of the European corn borer (ECB) are a textbook example in this respect. ‘Z’ females produce and ‘Z’ males preferentially respond to a ‘Z’ pheromone blend, while the ‘E’ race communicates via an ‘E’ blend. Both races do not freely hybridize in nature and their populations are genetically differentiated. A straightforward explanation would be that their reproductive isolation is a mere consequence of “assortative meeting” resulting from their different pheromones specifically attracting males towards same-race females at long range. However, previous laboratory experiments and those performed here show that even when moths are paired in a small box – i.e., when the meeting between sexual partners is forced – inter-race couples still have a lower mating success than intra-race ones. Hence, either the difference in attractivity of E vs. Z pheromones for males of either race still holds at short distance or the reproductive isolation between E and Z moths may not only be favoured by assortative meeting, but must also result from an additional mechanism ensuring significant assortative mating at close range. Here, we test whether this close-range mechanism is linked to the E/Z female sex pheromone communication system.Methodology/Principal FindingsUsing crosses and backcrosses of E and Z strains, we found no difference in mating success between full-sisters emitting different sex pheromones. Conversely, the mating success of females with identical pheromone types but different coefficients of relatedness to the two parental strains was significantly different, and was higher when their genetic background was closer to that of their male partner's pheromone race.Conclusions/SignificanceWe conclude that the close-range mechanism ensuring assortative mating between the E and Z ECB pheromone races is unrelated to the difference in female sex pheromone. Although the nature of this mechanism remains elusive, our results show that it is expressed in females, acts at close range, segregates independently of the autosome carrying Pher and of both sex chromosomes, and is widely distributed since it occurs both in France and in the USA.

More Rare Males in Ostrinia: Response of Asian Corn Borer Moths to the Sex Pheromone of the European Corn Borer

A previous flight tunnel study showed that 3-5% of European corn borer (ECB) moths, Ostrinia nubilalis (Z/E11-14:OAc), could fly upwind and make contact with sources releasing the sex pheromone of the related Asian corn borer (ACB), Ostrinia furnacalis (2:1 Z/E12-14:OAc). In this study, we show that rare males (3-4%) are also present in South Korean ACB that respond to the sex pheromone blends of the ECB UZ (97:3 Z/E11-14:OAc) and BE (1:99 Z/E11-14:OAc) pheromone races. We also show that the upwind flight response of a significant proportion of male ACB was antagonized by the addition of 1% Z9-14:OAc to the ACB blend, a compound that also antagonizes the upwind flight of ECB males. Male ACB flight behavior was not, however, affected by adding either of the ECB blends to the ACB blend, or by the addition of 50% 14:OAc, a compound identified from female pheromone glands of ACB and a number of other Ostrinia species. Additional flight tunnel tests with ACB to study the comparative aspects of ECB and ACB pheromone response specificity showed that male ACB exhibited maximal levels of upwind flight and source contact with doses of pheromone (30 and 100 microg on rubber septum sources) that also elicited maximal levels in the two ECB pheromone races. The maximal level of source contact for ACB (66%) was lower than observed with the UZ race of ECB to its pheromone blend (>95%), but comparable to those for the BE race of ECB (65-70%). Male ACB also flew upwind in high proportions to a broader range of ratios of Z/E12-14:OAc (80:20 to 20:80) than was previously observed for either of the ECB races.

Geographical Variation in Pheromone Response of the European Corn Borer, <I>Ostrinia nubilalis</I> (Lepidoptera: Crambidae), in North Carolina: A 20-Y Perspective

Abstract Pheromone traps were used to assess the distribution of two pheromone races of European corn borer, Ostrinia nubilalis (Hubner), in North Carolina, ≈10 and 20 yr after previous, similar assessments. In the previous studies, moths responding to a 97Z: 3E isomeric blend (Z blend) of 11-tetradecenyl acetate predominated in the far western parts of the state, whereas moths responding to a 3Z: 97E blend (E blend) prevailed in the east, with a substantial zone of overlap occurring in the eastern Piedmont. There was evidence that the E responsive population had expanded westward between 1978 and 1988. In this study, the distribution of the two races seemed to remain essentially unchanged from that observed in the late 1980s, and no evidence of a continued westward expansion of E responsive moths was detected.

Silent genes and rare males: A fresh look at pheromone blend response specificity in the European corn borer moth, Ostrinia nubilalis

The response of male moths from two pheromone races of the European corn borer, Ostrinia nubilalis, was measured in a flight tunnel assay to different ratios of structurally different compounds that comprise the sex pheromone of the Asian corn borer, Ostrinia furnacalis. For both O. nubilalis races, between 1 and 5% of the males completed upwind flights to two different blends of the O. furnacalis pheromone components (the 2:1 Z/E12-14:OAc female-produced blend, and a 97:3 Z/E mix), confirming that rare males exist in the O. nubilalis populations that can detect and respond to mixtures of the O. furnacalis pheromone components. Rare males that responded to the O. furnacalis blends also responded to their own O. nubilalis blends (97:3 or 1:99 Z/E11-14:OAc), indicating that rare O. nubilalis males are not preferentially sensitive to mixtures of the O. furnacalis compounds, but rather that they have a broad range of response specificity, which includes recognition of a wide range of conspecific female-produced ratios, and also recognition of heterospecific mixtures. The results support the hypothesis that saltational shifts in pheromone blend composition (Roelofs et al., 2002) can lead to the evolution of a new species-specific communication system, in part because the broad response specificity of some males includes the ability to respond in an agonistic manner to novel mixtures of compounds.

Incomplete barriers to mitochondrial gene flow between pheromone races of the North American pine engraver,Ips pini(Say) (Coleoptera, Scolytidae)

The pine engraver Ips pini (Say) is known to include three pheromone races, but gene flow between these races has not been investigated. We used maternally inherited mitochondrial DNA (mtDNA) variation to infer gene flow between 22 widely distributed North American populations of I. pini for a total of 217 individuals, based on 354 bp of the cytochrome oxidase I gene. Gene flow was estimated cladistically as migrants per generation (Nm) and as haplotype variation between populations (Nst). Three distinct mtDNA haplotype lineages, generally corresponding to eastern (I), Rocky Mountain (II) and western (III) regions of North America, were resolved with a total of 34 distinct I. pini haplotypes. The distributions of these lineages were largely congruent with the geographical ranges of the 'New York', 'California' and 'Idaho–Montana' pheromone races. Only individuals with lineage I mtDNA were observed among eastern populations, whereas individuals with lineage II or III mtDNA predominated among western populations. Gene flow (Nm and Nst) was generally moderate between all populations. However, the presence of lineage I mtDNA on the eastern side of western North America and the absence of lineage II and III mtDNA in eastern North America suggest directional gene flow from east to west. These results indicate that female-controlled assortative mating among pheromone races may disrupt gene flow between conspecifics, reflecting incomplete pre-mating barriers.