Gut bacteria mediate aggregation in the German cockroach.

Aggregation of the German cockroach, Blattella germanica, is regulated by fecal aggregation agents (pheromones), including volatile carboxylic acids (VCAs). We demonstrate that the gut microbial community contributes to production of these semiochemicals. Chemical analysis of the fecal extract of B. germanica revealed 40 VCAs. Feces from axenic cockroaches (no microorganisms in the alimentary tract) lacked 12 major fecal VCAs, and 24 of the remaining compounds were represented at extremely low amounts. Olfactory and aggregation bioassays demonstrated that nymphs strongly preferred the extract of control feces over the fecal extract of axenic cockroaches. Additionally, nymphs preferred a synthetic blend of 6 fecal VCAs over a solvent control or a previously identified VCA blend. To test whether gut bacteria contribute to the production of fecal aggregation agents, fecal aerobic bacteria were cultured, isolated, and identified. Inoculation of axenic cockroaches with individual bacterial taxa significantly rescued the aggregation response to the fecal extract, and inoculation with a mix of six bacterial isolates was more effective than with single isolates. The results indicate that the commensal gut microbiota contributes to production of VCAs that act as fecal aggregation agents and that cockroaches discriminate among the complex odors that emanate from a diverse microbial community. Our results highlight the pivotal role of gut bacteria in mediating insect-insect communication. Moreover, because the gut microbial community reflects the local environment, local plasticity in fecal aggregation pheromones enables colony-specific odors and fidelity to persistent aggregation sites.

The response to selection for broad male response to female sex pheromone and its implications for divergence in close-range mating behavior in the European corn borer moth, Ostrinia nubilalis.

Coordinated sexual communication systems, seen in many species of moths, are hypothesized to be under strong stabilizing natural selection. Stabilized communication systems should be resistant to change, but there are examples of species/populations that show great diversification. A possible solution is that it is directional sexual selection on variation in male response that drives evolution. We tested a component of this model by asking whether 'rare' males (ca. 5 % of all males in a population) of the European corn borer moth (ECB), Ostrinia nubilalis, that respond to the sex pheromones of both ECB and a different Ostrinia species (O. furnacalis, the Asian corn borer, ACB), might play an important role in diversification. We specifically tested, via artificial selection, whether this broad male response has an evolvable genetic component. We increased the frequency of broad male response from 5 to 70 % in 19 generations, showing that broad-responding males could be important for the evolution of novel communication systems in ECB. We did not find a broader range of mating acceptance of broad males by females of the base population, however, suggesting that broad response would be unlikely to increase in frequency without the involvement of other factors. However, we found that ECB selection-line females accepted a broader range of courting males, including those of ACB, than did females of the base population. Thus, a genetic correlation exists between broad, long-range response to female sex pheromone and the breadth of female acceptance of males at close range. These results are discussed in the context of evolution of novel communication systems in Ostrinia.

Comparing peripheral olfactory coding with host preference in the rhagoletis species complex.

Recent studies have shown that flies from sympatric populations of Rhagoletis pomonella infesting hawthorn, apple, and flowering dogwood fruit can distinguish among unique volatile blends identified from each host. Analysis of peripheral chemoreception in Rhagoletis flies suggests that changes in receptor specificity and/or receptor neuron sensitivity could impact olfactory preference among the host populations and their hybrids. In an attempt to validate these claims, we have combined flight tunnel analyses and single sensillum electrophysiology in F(2) and backcross hybrids displaying a variety of behavioral phenotypes. Results show that differences in peripheral chemoreception among second-generation adults do not provide a direct correlation between peripheral coding and olfactory behavior. We conclude that either the plasticity of the central nervous system in Rhagoletis can compensate for significant alterations in peripheral coding or that peripheral changes present subtle effects on behavior not easily detectable with current techniques. The results of this study imply that the basis for olfactory behavior in Rhagoletis has a complicated genetic and neuronal basis, even for populations with a recent divergence in preference.

Sex pheromone of the scarab beetle Phyllophaga (Phytalus) georgiana (horn).

The sex pheromone of Phyllophaga (Phytalus) georgiana was characterized as valine methyl ester, tentatively the L-enantiomer. This is the first sex pheromone identified from the Phyllophaga subgenus Phytalus. The pheromone was extracted from female glands, the active component isolated by coupled gas chromatography-electroantennogram detection analysis, characterized by mass spectrometry, and shown to be active in field tests. The seasonal flight pattern was determined for P. georgiana as well as for three other species, P. anxia (both northern and southern genitalic forms), P. gracilis, and P. postrema. The latter three species were captured in traps baited with L-isoleucine methyl ester.

Olfactory neuron responsiveness and pheromone blend preference in hybrids between Ostrinia furnacalis and Ostrinia nubilalis (Lepidoptera: Crambidae).

The olfactory receptor neuron (ORN) and behavioral responses of hybrids between the Asian corn borer (ACB), Ostrinia furnacalis, and the E-strain European corn borer (ECB(E)), Ostrinia nubilalis were examined and compared to the parental populations. In hybrids and both parents, the large-spike-size ORN was capable of responding to all four pheromone components of ACB and ECB, despite differences in which compounds elicited the greatest spike frequency in each population. There was a small-spiking ORN more narrowly tuned to the minor pheromone components in both ACB and ECB(E). In hybrids the homologous small-spiking ORN was tuned primarily to the ECB(E) minor pheromone component, with some responsiveness to the ACB minor component. Both species and all the hybrids had an intermediate spike-size ORN tuned primarily to their common behavioral antagonist. Dominance of responsiveness to the ECB(E) versus the ACB minor pheromone component on the small-spiking ORN may explain the greater tendency of hybrids to fly upwind to the ECB(E) pheromone blend than the ACB blend. This finding points toward a distinct evolutionary role for this ORN in allowing a pheromone shift.

Altered olfactory receptor neuron responsiveness in rare Ostrinia nubilalis males attracted to the O. furnacalis pheromone blend.

Three percent of E-strain Ostrinia nubilalis males fly upwind in response to the Ostrinia furnacalis pheromone blend [a 40:60 ratio of (E)-12-tetradecenyl acetate to (Z)-12-tetradecenyl acetate (E12-14:OAc to Z12-14:OAc)], in addition to their own pheromone blend [a 99:1 ratio of (E)-11-tetradecenyl acetate to (Z)-11-tetradecenyl acetate) (E11-14:OAc to Z11-14:OAc)]. We assessed the olfactory receptor neuron (ORN) responses of these behaviorally "rare" males versus those of normal males. For the three ORNs housed within each sensillum, we tested responsiveness to Z12-14:OAc, E12-14:OAc, Z11-14:OAc, E11-14:OAc, and the behavioral antagonist (Z)-9-tetradecenyl acetate (Z9-14:OAc). Z11-14:OAc, E11-14:OAc, and Z9-14:OAc stimulated ORNs exhibiting distinct small, large, and medium spike sizes, respectively. For rare and normal males, both Z12-14:OAc and E12-14:OAc usually elicited responses from the largest-spiking ORN. In many ORNs of normal males, Z12-14:OAc or E12-14:OAc stimulated the smaller-spiking ORN that is responsive to Z11-14:OAc. In rare males, detectable ORN responses from the smaller-spiking ORN in response to Z12- and E12-14:OAc were virtually non-existent. These differences in ORN tuning in rare males will tend to create an ORN firing ratio between the large- and small-spiking ORNs in response to the O. furnacalis blend that is similar to that elicited by the O. nubilalis blend.

Evidence of olfactory antagonistic imposition as a facilitator of evolutionary shifts in pheromone blend usage in Ostrinia spp. (Lepidoptera: Crambidae).

Olfactory receptor neuron (ORN) response was measured to assess why some males ("rare males") of the Asian corn borer (ACB), Ostrinia furnacalis, have a broad behavioral response to fly upwind to both the ACB and the European corn borer (ECB), Ostrinia nubilalis, pheromone blends. We performed single-sensillum electrophysiological recordings on ACB males that had been behaviorally assessed for upwind flight response to the ACB blend [60:40 (Z)-12-tetradecenyl acetate (Z12-14:OAc) to (E)-12-tetradecenyl acetate (E12-14:OAc)], as well as to ECB (Z-strain) and ECB (E-strain) blends [3:97 and 99:1 (Z)-11-tetradecenyl acetate (Z11-14:OAc) to (E)-11-tetradecenyl acetate (E11-14:OAc)]. Sensilla from all types of males had large- and small-spike-sized ORNs responding strongly to Z12- or E12-14:OAc, but weakly to Z11- and E11-14:OAc. In the majority of males ("normal males") that flew upwind only to the ACB blend, Z11-14:OAc elicited responses in an intermediate spike-sized ORN associated with behavioral antagonism that is mainly tuned to (Z)-9-tetradecenyl acetate (Z9-14:OAc). In the rare-type ACB males that flew to both the ACB and ECB pheromone blends, Z11-14:OAc did not stimulate this ORN. Increased responsiveness to ancestral pheromone components by ORNs associated with behavioral antagonism could be instrumental in reproductive character displacement, or in reinforcement and reproductive isolation during speciation by helping to increase assortative mating between males and females in derived populations that use novel sex pheromone blends.

Molecular genetics and evolution of pheromone biosynthesis in Lepidoptera.

A GREAT DIVERSITY OF PHEROMONE STRUCTURES ARE USED BY MOTH SPECIES (INSECTA: Lepidoptera) for long-distance mating signals. The signal/response channel seems to be narrow for each species, and a major conundrum is how signal divergence has occurred in the face of strong selection pressures against small changes in the signal. Observations of various closely related and morphologically similar species that use pheromone components biosynthesized by different enzymes and biosynthetic routes underscore the question as to how major jumps in the biosynthetic routes could have evolved with a mate recognition system that is based on responses to a specific blend of chemicals. Research on the desaturases used in the pheromone biosynthetic pathway for various moth species has revealed that one way to make a major shift in the pheromone blend is by activation of a different desaturase from mRNA that already exists in the pheromone gland. Data will be presented to support the hypothesis that this process was used in the evolution of the Asian corn borer, Ostrinia furnacalis species. In that context, moth sex-pheromone desaturase genes seem to be evolving under a birth-and-death process. According to this model of multigene family evolution, some genes are maintained in the genome for long periods of time, whereas others become deleted or lose their functionality, and new genes are created through gene duplication. This mode of evolution seems to play a role in moth speciation, as exemplified by the case of the Asian corn borer and European corn borer, Ostrinia nubilalis species.

Effects of egg-to-adult development time and adult age on olfactory neuron response to semiochemicals in European corn borers.

We used the cut-sensillum technique to assess the effect of both adult age and egg-to-adult development time on olfactory neuron responses of Z strain moths of the European corn borer, Ostrinia nubilalis. Compounds tested included the pheromone components, (Z)-11-tetradecenyl acetate and (E)-11-tetradecenyl acetate, the behavioral antagonist, (Z)-9-tetradecenyl acetate, and components of the O. furnicalis (Asian corn borer) sex pheromone, (Z)-12-tetradecenyl acetate and (E)-12-tetradecenyl acetate. The proportion of moths having neurons responding to the two O. nubilalis sex pheromone components and antagonist increased with longer development time and age. The spike frequency of neurons responding to (E)-11-tetradecenyl acetate and the antagonist increased with longer development time. Fourteen of 45 moths with neurons sensitive to either of the O. nubilalis pheromone components responded to (Z)-12-tetradecenyl acetate or (E)-12-tetradecenyl acetate. The likelihood of (Z)-12-tetradecenyl acetate stimulating a neuron similar in spike shape and waveform to that responding to (E)-11-tetradecenyl acetate increased with development time.

Trapping Phyllophaga spp. (Coleoptera: Scarabaeidae: Melolonthinae) in the United States and Canada using sex attractants.

The sex pheromone of the scarab beetle, Phyllophaga anxia, is a blend of the methyl esters of two amino acids, L-valine and L-isoleucine. A field trapping study was conducted, deploying different blends of the two compounds at 59 locations in the United States and Canada. More than 57,000 males of 61 Phyllophaga species (Coleoptera: Scarabaeidae: Melolonthinae) were captured and identified. Three major findings included: (1) widespread use of the two compounds [of the 147 Phyllophaga (sensu stricto) species found in the United States and Canada, males of nearly 40% were captured]; (2) in most species intraspecific male response to the pheromone blends was stable between years and over geography; and (3) an unusual pheromone polymorphism was described from P. anxia. Populations at some locations were captured with L-valine methyl ester alone, whereas populations at other locations were captured with L-isoleucine methyl ester alone. At additional locations, the L-valine methyl ester-responding populations and the L-isoleucine methyl ester-responding populations were both present, producing a bimodal capture curve. In southeastern Massachusetts and in Rhode Island, in the United States, P. anxia males were captured with blends of L-valine methyl ester and L-isoleucine methyl ester.

Desaturases from the spotted fireworm moth (Choristoneura parallela) shed light on the evolutionary origins of novel moth sex pheromone desaturases.

Six acyl-CoA desaturase-encoding cDNAs from mRNA isolated from the spotted fireworm moth, Choristoneura parallela (Lepidoptera: Tortricidae) were characterized and assayed for functionality. The expression levels of these cDNAs were determined in the pheromone gland and fat body by real-time PCR and the resulting patterns are in line with results from published studies on other moth sex pheromone desaturases. The cDNAs were found to correspond to six genes. Using both biochemical and phylogenetic analyses, four of these were found to belong to previously characterized desaturase functional groups [the Delta 10,11, the Delta 9 (16>18) and the Delta 9 (18>16) groups]. A desaturase highly expressed in the pheromone gland was a novel E11 desaturase that was specific to 14-carbon precursor acids. The fifth gene [CpaZ9(14-26)] was found to display a novel Z9 activity indicating that it belongs to a new Delta 9 functional group, whereas the sixth gene was determined to be nonfunctional with respect to desaturase activity. In accordance with previous studies, we find that desaturases of the Delta 10,11 and Delta 14 groups, which are the fastest evolving desaturases and possess the novel pheromone biosynthetic function, are expressed primarily in the pheromone gland whereas all other desaturases, which do not possess the novel reproductive function, evolve more slowly and display the ancestral metabolic function and pattern of gene expression.

Moth desaturase characterized that produces both Z and E isomers of delta 11-tetradecenoic acids.

The redbanded leafroller moth, Argyrotaenia velutinana (Lepidoptera: Tortricidae) uses a 92:8 mixture of (Z)-11- and (E)-11-tetradecenyl acetate in its pheromone blend. These are produced in the abdominal pheromone gland from the corresponding acids, which are biosynthesized in the gland in a 3:2 Z/E ratio by desaturation of myristoyl CoA. The delta 11 desaturase involved in this reaction exhibits unusual substrate and stereospecificities in specifically producing Z11 and E11 isomers of tetradecenoic acid, and exhibiting no activity with C16 and C18 precursor acids. This report describes the cloning and expression of the redbanded leafroller moth delta 11 desaturase, and compares its amino-acid sequence to those of other known insect Z9, Z10, Z11, and E11 desaturases. The metabolic Z9 desaturase from fat body tissue also was cloned and expressed, and found mainly to produce Z9-16:Acid and Z9-18:Acid. The open reading frame of the delta 11 desaturase encodes a protein with 329 amino acids, whereas the open reading frame of the Z9 desaturase encodes a protein with 351 amino acids. Addition of this new delta 11 desaturase with its different substrate and regiospecificites to the databank of characterized integral-membrane desaturases will be key in efforts to determine amino-acid mutations responsible for the wide array of unsaturated fatty-acid products.

Expression and evolution of delta9 and delta11 desaturase genes in the moth Spodoptera littoralis.

Desaturation of fatty acids is a key reaction in the biosynthesis of moth sex pheromones. The main component of Spodoptera littoralis sex pheromone blend is produced by the action of Delta11 and Delta9 desaturases. In this article, we report on the cloning of four desaturase-like genes in this species: one from the fat body (Sls-FL1) and three (Sls-FL2, Sls-FL3 and Sls-FL4) from the pheromone gland. By means of a computational/phylogenetic method, as well as functional assays, the desaturase gene products have been characterized. The fat body gene expressed a Delta9 desaturase that produced (Z)-9-hexadecenoic and (Z)-9-octadecenoic acids in a (1:4.5) ratio, whereas the pheromone gland Sls-FL2 expressed a Delta9 desaturase that produced (Z)-9-hexadecenoic and (Z)-9-octadecenoic acids in a (1.5:1) ratio. Although both Delta9 desaturases produced (Z)-9-tetradecenoic acid from myristic acid, transformed yeast grown in the presence of a mixture of myristic and (E)-11-tetradecenoic acids produced (Z,E)-9,11-tetradecadienoic acid, but not (Z)-9-tetradecenoic acid. The Sls-FL3 gene expressed a protein that produced a mixture of (E)-11-tetradecenoic, (Z)-11-tetradecenoic, (Z)-11-hexadecenoic and (Z)-11-octadecenoic acids in a 5:4:60:31 ratio. Despite having all the characteristics of a desaturase gene, no function could be found for Sls-FL4.

Characterization of Z/E11- and Z9-desaturases from the obliquebanded leafroller moth, Choristoneura rosaceana.

A (triangle up) 11-desaturase gene was cloned from the sex pheromone gland of the obliquebanded leafroller moth, Choristoneura rosaceana. The desaturase cDNA sequence spans 1300 nucleotides with an open reading frame encoding a 335 amino-acid protein, which has 81% identity to a Z/E11-desaturase of the redbanded leafroller moth, Argyrotaenia velutinana. A functional assay with a pYES2 yeast expression system demonstrated that the (triangle up) 11-desaturase exhibits unusual substrate and stereospecificities in producing a Z/E11 mixture (7:1) of only C14 acids. A metabolic Z9-desaturase also was cloned from fat body of this species, and proved to be in the class that produces more Z9-16:Acid than Z9-18:Acid.

Influence of trap design on upwind flight behavior and capture of female grape berry moth (Lepidoptera: Tortricidae) with a kairomone lure.

Oil-coated clear panel traps baited with a host plant-based kairomone lure have successfully been used for monitoring female grape berry moth, Paralobesia viteana (Clemens) (Lepidoptera: Tortricidae), but low capture rates as well as difficulty in servicing these traps makes them unsuitable for commercial use. We compared the performance of different trap designs in a flight tunnel and in a vineyard by using a 7-component synthetic kairomone blend, with a focus on trap visual cues. In flight tunnel experiments, a clear delta trap performed better than other traps. When we tested clear delta, green delta, or clear wing traps baited with a cut grape shoot, >50% of female grape berry moths made complete upwind flights. However, the clear delta trap was the only design that resulted in female moths entering the trap. Similar results were observed when females were tested with different traps (clear delta, green delta, white delta, clear wing, or green wing traps) baited with the kairomone lure. Adding a visual pattern that mimicked grape shoots to the outside surface of the clear delta trap resulted in 66% of the females that made upwind flights entering the trap. However, the positive effect of adding a visual pattern to the trap was not observed in a vineyard setting, where clear delta traps with or without a visual pattern caught similar numbers of females. Still, the number of male and female grape berry moths captured in clear delta traps with or without a visual pattern was not significantly different from the number of male and female grape berry moths captured in panel traps, suggesting that the use of these delta traps could be a less cumbersome alternative to oil-coated panel traps for monitoring female grape berry moth.

Eavesdropping on plant volatiles by a specialist moth: significance of ratio and concentration.

We investigated the role that the ratio and concentration of ubiquitous plant volatiles play in providing host specificity for the diet specialist grape berry moth Paralobesia viteana (Clemens) in the process of locating its primary host plant Vitis sp. In the first flight tunnel experiment, using a previously identified attractive blend with seven common but essential components ("optimized blend"), we found that doubling the amount of six compounds singly [(E)- & (Z)-linalool oxides, nonanal, decanal, ß-caryophyllene, or germacrene-D], while keeping the concentration of other compounds constant, significantly reduced female attraction (average 76% full and 59% partial upwind flight reduction) to the synthetic blends. However, doubling (E)-4,8-dimethyl 1,3,7-nonatriene had no effect on female response. In the second experiment, we manipulated the volatile profile more naturally by exposing clonal grapevines to Japanese beetle feeding. In the flight tunnel, foliar damage significantly reduced female landing on grape shoots by 72% and full upwind flight by 24%. The reduction was associated with two changes: (1) more than a two-fold increase in total amount of the seven essential volatile compounds, and (2) changes in their relative ratios. Compared to the optimized blend, synthetic blends mimicking the volatile ratio emitted by damaged grapevines resulted in an average of 87% and 32% reduction in full and partial upwind orientation, respectively, and the level of reduction was similar at both high and low doses. Taken together, these results demonstrate that the specificity of a ubiquitous volatile blend is determined, in part, by the ratio of key volatile compounds for this diet specialist. However, P. viteana was also able to accommodate significant variation in the ratio of some compounds as well as the concentration of the overall mixture. Such plasticity may be critical for phytophagous insects to successfully eavesdrop on variable host plant volatile signals.

Monitoring grape berry moth (Paralobesia viteana: Lepidoptera) in commercial vineyards using a host plant based synthetic lure.

For some Lepidopteran pests, such as the grape berry moth Paralobesia viteana (Clemens), poor correlation between males captured in traps baited with sex pheromone and oviposition activities of female moths has called into question the value of pheromone-based monitoring for these species. As an alternative, we compared the capture of female and male grape berry moth in panel traps baited with synthetic host volatiles with captures of males in pheromone-baited wing traps over two growing seasons in two blocks of grapes in a commercial vineyard in central New York. Lures formulated in hexane to release either 7-component or 13-component host volatile blends captured significantly more male and female grape berry moth on panel traps compared with the numbers captured on panel traps with hexane-only lures. For both sexes over both years, the same or more moths were captured in panel traps along the forest edge compared with the vineyard edge early in the season but this pattern was reversed by mid-season. Male moths captured in pheromone-baited wing traps also displayed this temporal shift in location. There was a significant positive correlation between captured males and females on panel traps although not between females captured on panel traps and males captured in pheromone-baited traps for both years suggesting pheromone traps do not accurately reflect either female or male activity. Male moths captured in pheromone traps indicated a large peak early in each season corresponding to first flight followed by lower and variable numbers that did not clearly indicate second and third flights. Panel trap data, combining males and females, indicated three distinct flights, with some overlap between the second and third flights. Peak numbers of moths captured on panel traps matched well with predictions of a temperature-based phenology model, especially in 2008. Although effective, panel traps baited with synthetic host lures were time consuming to deploy and maintain and captured relatively few moths making them impractical, in the current design, for commercial purposes.

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 F(1) 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 F(1) 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.

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.

Flight tunnel responses of female grape berry moth (Paralobesia viteana) to host plants.

Semiochemicals play important roles in mate and host recognition of herbivorous insects, such as moths, and flight tunnels have been an effective tool in the identification of these bioactive compounds. However, more work has been carried out on pheromones than on host plant cues, and few examples exist where flight tunnel evaluations of host cues have resulted in a lure that is attractive under field conditions. Our goal was to determine whether the flight tunnel could be used to evaluate the response of a specialist moth, grape berry moth (GBM), to its host plant (grapevines), by incorporating ecological and physiological aspects of GBM biology. We found grape shoot tips and mature leaves were more attractive to female GBM than unripe and ripe berries or flowers. Under optimized flight tunnel conditions, approximately 80% of tested females flew upwind and closely approached or landed on the most preferred target. Mating status, wind speed, the time of day, and the presence/absence of patterns that resemble grape tissues on the top of the flight tunnel all significantly affected the responses of female GBM. Consideration of these factors in flight tunnel assays will aid in the development of a synthetic lure that can be used to monitor female moths in the field.