Peter Mayo

Evidence for a Volatile Pheromone in Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) that Increases Attraction to a Host Foliar Volatile

ABSTRACT Analysis by gas chromatography/mass spectrometry (GC/MS) of volatiles from virgin female emerald ash borer, Agrilus planipennis Fairmaire confirmed the emission of (3Z)-lactone [(3Z)-dodecen-12-olide] but not its geometric isomer, (3E)-lactone [(3E)-dodecen-12-olide]. Gas chromatographic/electroantennographic (GC/EAD) analysis of synthetic (3Z)-lactone, which contained 10% (3E) -lactone, showed a strong response of male and female antennae to both isomers. EAG analysis with 0.01 - to100-µg dosages showed a positive dose response, with females giving significantly higher responses than males. In field experiments with sticky purple prism traps, neither lactone isomer affected catches when combined with ash foliar or cortical volatiles (green leaf volatiles or Phoebe oil, respectively). However, on green prism traps, the (3Z)-lactone significantly increased capture of male A. planipennis when traps were deployed in the canopy. Captures of males on traps with both (3E)-lactone and (3Z)-hexenol or with (3Z)-lactone and (3Z)-hexenol were increased by 45–100%, respectively, compared with traps baited with just (3Z)-hexenol. In olfactometer bioassays, males were significantly attracted to (3E) -lactone, but not the (3Z) -lactone or a 60:40 (3E): (3Z) blend. The combination of either (3E)- or (3Z)-lactone with Phoebe oil was not significantly attractive to males. Males were highly attracted to (3Z)-hexenol and the (3Z)-lactone + (3Z)-hexenol combination, providing support for the field trapping results. These data are the first to demonstrate increased attraction with a combination of a pheromone and a green leaf volatile in a Buprestid species.

Attraction of Agrilus planipennis (Coleoptera: Buprestidae) to a Volatile Pheromone: Effects of Release Rate, Host Volatile, and Trap Placement

ABSTRACT Attraction of emerald ash borer, Agrilus planipennis Fairmaire, to a volatile pheromone was demonstrated in three field experiments using baited green sticky traps. A dose-response curve was generated for male A. planipennis to increasing release rates of (3Z)-dodecen-12-olide ((3Z)lactone) in combination with the green leaf volatile, (3Z)-hexenol. Only the lowest release rate (<2.50 µg/d) of (3Z)-lactone significantly increased captures of male A. planipennis, as compared with traps baited with (3Z)-hexenol alone. Effect of trap height, (3Z)-lactone, and (3Z)-hexenol and their interactions on the trap capture of A. planipennis was determined in a factorial experiment. Number of males per trap was significantly and positively affected by (3Z)-lactone, (3Z)-hexenol, and trap height whereas number of females per trap was affected by trap height only; none of the interactions were significant. As predicted, the greatest mean catch of males was in traps baited with (3Z)-lactone and (3Z)-hexenol placed high in the canopy. Electroantennogram tests on the bark volatile, 7-epi-sesquithujene, demonstrated the ability of male and female A. planipennis antennae to detect and respond to this compound, particularly the (+)-7-epi-sesquithujene isomer. Results from an olfactometer bioassay and field testing did not demonstrate attraction of either males or females to (+)-7-epi-sesquithujene. These data increase our understanding of the pheromone ecology of the invasive emerald ash borer, provide further confirmation of the behavioral activity of the female-produced lactone pheromone, and should increase the ability to detect A. planipennis infestations where they are present.

Contact Sex Pheromones Identified for Two Species of Longhorned Beetles (Coleoptera: Cerambycidae) Tetropium fuscum and T. cinnamopterum in the Subfamily Spondylidinae

ABSTRACT Male Tetropium fuscum (F.) and T. cinnamopterum Kirby mated with live and dead (freeze-killed) conspecific females upon antennal contact, but did not respond to dead females after cuticular waxes were removed by hexane rinsing. Significantly fewer males of each species attempted to copulate with live or dead heterospecific females than with conspecifics, indicating that mate recognition was mediated by species-specific contact sex pheromones in the females cuticular hydrocarbons. GC/MS analysis of T. fuscum elytra identified n-alkanes and mono-methyl branched alkanes of which 11-methylheptacosane and 3- and 5-methyltricosanes were dominant in females. Full male responses, including copulatory behavior, were restored with application of enantiomerically pure synthetic (S)-11-methyl-heptacosane at 40 µg /female (one female equivalent) but not with racemic or (R)-11-methyl-heptacosane. The cuticular hydrocarbons on T. cinnamopterum elytra included 11-methyl-heptacosane as well as n-alkanes, methyl-branched alkanes, mono-alkenes, and (Z, Z)-6, 9-alkadienes. (Z)-9-pentacosene, (Z)-9-heptacosene, and 11-methyl-heptacosane were female dominant, but only (Z)-9-pentacosene elicited precopulatory behaviors in conspecific males at levels similar to those behaviors elicited by unrinsed females, but elicited copulation in fewer than half of males. At female equivalent dosages (10 µg), neither (Z)-9-heptacosene nor (S)-11-methylheptacosane elicited responses in males that were significantly different from those responses to a rinsed female but when applied together, the proportion of males responding was significantly increased. 11-methyl-heptacosene is thus a contact pheromone component common to both species, which may explain the heterospecific mating attempts by some males.

Steps in the Biosynthesis of Fuscumol in the Longhorn Beetles Tetropium fuscum (F.) and Tetropium cinnamopterum Kirby

Fuscumol [(2S,5E)-6,10-dimethyl-5,9-undecadien-2-ol] was recently identified as the male-produced aggregation pheromone of the brown spruce longhorn beetle, Tetropium fuscum (F.), and the eastern larch borer, Tetropium cinnamopterum Kirby. Several other species use this homoterpenoid alcohol motif, its ketone, or its acetate as part of their pheromone system. Investigation of the biosynthesis of this compound in these two Tetropium species demonstrated that geranylacetone [(5E)-6,10-dimethyl-5,9-undecadien-2-one] and farnesol [(2E,6E)-3,7,11-trimethyl-2,6,10-dodecatrien-1-ol] are both intermediates in this process. This was accomplished by applying deuterium-labeled geranylacetone and deuterium-labeled farnesol in separate experiments to the abdominal sterna of live T. fuscum and T. cinnamopterum and analyzing the deuterium labeling in the fuscumol and geranylacetone emitted by the insects with solid-phase microextraction (SPME) and GC/MS analysis. Deuterium labeling studies also showed that nerolidol[(3S,6E)-3-hydroxy-3,7,11-trimethyl-1,6,10-dodecatriene] and 2,3-epoxyfarnesol are not intermediates in fuscumol or geranylacetone synthesis in T. fuscum or T. cinnamopterum. Tissue-specific expression of T. fuscum farnesyl diphosphate synthase (TfFPPS), an enzyme expected to provide a key fuscumol precursor, was measured. TfFPPS transcripts were relatively abundant in male midguts, but were also present at significant levels in other tissues.

A biologically active analog of the sex pheromone of the emerald ash borer, Agrilus planipennis.

The emerald ash borer, Agrilus planipennis (Coleoptera: Buprestidae) (EAB), is an invasive species causing unprecedented levels of mortality to ash trees in its introduced range. The female-produced sex pheromone of EAB has been shown to contain the macrocyclic lactone (3Z)-dodecen-12-olide. This compound and its geometrical isomer, (3E)-dodecen-12-olide, have been demonstrated previously to be EAG active and, in combination with a host-derived green leaf volatile, (3Z)-hexenol, to be attractive to male EAB in green prism traps deployed in the ash tree canopy. In the current study, we show that the saturated analog, dodecan-12-olide, is similarly active, eliciting an antennal response and significant attraction of EAB in both olfactometer and trapping bioassays in green traps with (3Z)-hexenol. Conformational modeling of the three lactones reveals that their energies and shapes are very similar, suggesting they might share a common receptor in EAB antennae. These findings provide new insight into the pheromone ecology of this species, highlighting the apparent plasticity in response of adults to the pheromone and its analog. Both of the unsaturated isomers are costly to synthesize, involving multistep, low-yielding processes. The saturated analog can be made cheaply, in high yield, and on large scale via Mitsunobu esterification of a saturated ω-hydroxy acid or more simply by Baeyer-Villiger oxidation of commercially available cyclododecanone. The analog can thus provide an inexpensive option as a lure for detection surveys as well as for possible mitigation purposes, such as mating disruption.

Effects of Pheromone Release Rate and Trap Placement on Trapping of Agrilus planipennis (Coleoptera: Buprestidae) in Canada

ABSTRACT The emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), is a devastating insect pest in its introduced range. A trapping system that increases trap catches or detection rates in low-density populations would be beneficial for survey programs. Five trapping experiments were conducted to investigate factors influencing capture rates of male beetles on dark green traps baited with the A. planipennis pheromone, (3Z)-dodecen-12-olide ((3Z)-lactone), plus the green leaf volatile, (3Z)-hexenol. Low doses (0.001–1.0 mg) of (3Z)-lactone + (3Z)-hexenol did not consistently increase captures of male A. planipennis. In other experiments, mean captures of males were significantly higher on traps baited with a moderate dose (3.0 mg/septum) of (3Z)-lactone + (3Z)-hexenol, compared with lower doses (0.001, 0.1, and 1.0 mg) or (3Z)-hexenol alone. Next, we demonstrated that addition of (3Z)-lactone to traps baited with (3Z)-hexenol resulted in significantly greater increases in male captures when pairs of traps were placed on the same tree, than when traps were placed on adjacent trees. Moreover, significantly more A. planipennis were captured on pheromone-baited traps placed in the southern versus northern aspect of the crown. These results highlight the importance of experimental set-up for elucidating lure treatment effects and also suggests the (3Z)-lactone may be more active at close range. Our findings increase our understanding of the pheromone ecology of this species and lend support toward the use of dark green traps baited with 3.0 mg (3Z)-lactone + (3Z)-hexenol deployed in the south aspect of the canopy for detection programs for this insect.

Developing trapping protocols for wood-boring beetles associated with broadleaf trees

Longhorn and jewel beetles are often moved intercontinentally within woody materials. The common use of hardwoods in solid wood-packaging requires efficient trapping protocols for broadleaf-associated species. We tested the effect of lure (ethanol vs multi-lure), trap color (green vs purple), and trap height (understory vs canopy) on the longhorn and jewel beetle species trapped in multi-funnel traps set up in both seminatural forests and reforested forests in Italy. Traps were deployed in a 2 × 2 × 2 factorial scheme, and the eight different treatments were replicated 17 times in a randomized complete block design, in which each block was a different site. Thirty-five longhorn beetle species (2 non-native) and 15 jewel beetle species (all native) were trapped. The multi-lure was more effective than ethanol at detecting most longhorn beetles at both the species and subfamily level (except Lepturinae), but had no effect on the detection of jewel beetles. Trap color affected both jewel (green better than purple) and longhorn beetles with mixed responses among subfamilies. Species richness and/or abundance of both families was greater in the canopy than the understory, but trends were more heterogeneous at lower taxonomic levels (i.e., significant effect on Cerambycinae and Lepturinae but not on Lamiinae). In general, we showed that green multi-funnel traps baited with multi-lure, and setup in the canopy may be an efficient trapping protocol for European longhorn and jewel beetles associated with broadleaf trees. This information can increase efficacy of early-detection programs carried out both inside and outside of Europe.

Use of contact chemical cues in prey discrimination by Cerceris fumipennis

Cerceris fumipennis Say (Hymenoptera: Crabronidae) uses a wide range of adult buprestid beetles to provision nests. Wasps seldom make ‘mistakes’ and attack beetles in other families. Bioassays showed that the wasps use contact chemical cues in the beetles’ epicuticle to discriminate buprestids from other beetles. Wasps rejected buprestids that had their cuticular hydrocarbons (CHC) removed by being washed in solvents. The washed, rejected buprestids became acceptable to 70% of wasps when they were coated with an extract of buprestids’ own CHC. Washed buprestids coated with extracts of non‐buprestid beetle CHC were not accepted. Analyses of buprestid CHC profiles showed that they are relatively simple, consisting of five classes. Other coleopteran families that may co‐occur in the arboreal hunting habitat of C. fumipennis have a broader range of CHC classes. Experiments adding buprestid hydrocarbons to a non‐buprestid beetle, unwashed Popillia japonica Newman (Coleoptera: Scarabaeidae), suggested that wasps may be deterred by the presence of CHC classes not found in Buprestidae. Adding a synthesized dimethyl‐branched hydrocarbon, a class of CHC found in Chrysomelidae but not Buprestidae, to the cuticle of unwashed buprestid beetles, caused wasps to reject the buprestid beetles. We propose that CHC act as a kairomone for C. fumipennis, triggering attack, whereas classes of CHC not found in Buprestidae disrupt this response, and thus help to determine the prey range of the wasp.

Concise Synthesis of (3Z)-Dodecen-12-olide, Pheromone Component of the Emerald Ash Borer

Abstract Emerald ash borer (EAB), Agrilus planipennis Fairmaire, is an invasive insect that has killed millions of ash trees in Canada and the USA. (3Z)-Dodecen-12-olide is a known female-produced pheromone of this insect, and a concise, three-step synthesis of a 2:1 blend of (3Z)-dodecen-12-olide and (3E)-dodecen-12-olide starting from commercially available (2-carboxyethyl)triphenylphosphonium bromide and 10-bromo-1-decene is described. The key steps in this synthesis are a lithium-salt-free Wittig reaction and an intramolecular SN2 esterification. Both of these macrocyclic lactones are behaviorally active toward EAB, and the 2:1 blend whose synthesis is described here has the potential to be a detection agent, mating disruptor, or mass trapping agent, which could be used in the control of EAB. GRAPHICAL ABSTRACT

Synthesis of (3E)-Dodecen-12-olide, a Potential Pheromone Component of the Emerald Ash Borer

Abstract Emerald ash borer (EAB), Agrilus planipennis Fairmaire, is an invasive insect that has killed millions of ash trees in the USA and Canada. A concise synthesis of a potential EAB pheromone component, (3E)-dodecen-12-olide, using highly stereoselective Julia–Kocienski olefination as the key step, is reported. Supplemental materials are available for this article. Go to the publishers online edition of Synthetic Communications® to view the free supplemental file. GRAPHICAL ABSTRACT