Gnathotrichus sulcatus: synergistic response to enantiomers of the aggregation pheromone sulcatol.

In laboratory and field bioassays, Gnathotrichus sulcatus responded to sulcatol (6-methyl-5-hepten-2-ol) only when both enantiomers were present. Response was greater to racemic sulcatol than to a mixture (65 : 35) of S-(+) and R-(-) enantiomers, the naturally occurring isomeric ratio. Enantiomer-specific active sites on receptor proteins in the same or different cells are implicated.

Specific biochemical marker-based techniques for the identification of damage to Douglas-fir seed resulting from feeding by the western conifer seed bug, Leptoglossus occidentalis Heidemann (Hemiptera: Coreidae).

Specific biochemical marker-based techniques were tested for their ability to distinguish between seeds of Douglas-fir, Pseudotsuga menziesii (Mirbel) Franco, that were filled or unfilled (aborted) at maturity and those that were damaged or emptied by the western conifer seed bug, Leptoglossus occidentalis Heidemann. A polyclonal antibody raised against salivary gland extracts from L. occidentalis successfully identified residual salivary proteins on Western blots containing proteins from Douglas-fir seeds that had sustained various degrees of seed bug feeding damage. In a single blind experiment, the polyclonal antibody correctly identified 100% of undamaged control, 97% of unfilled control (aborted), and 98% of seed bug damaged seeds. Polyclonal antibodies raised against insoluble alfalfa crystalloid storage protein (11S globulin) detected the depletion of 11S globulin and the subsequent appearance of its hydrolyzed fragments in the soluble protein fraction of Douglas-fir seeds that were fed-upon by the seed bug. Feeding by L. occidentalis nymphs caused ca. 98% depletion of insoluble protein, but only ca. 53% reduction in the amount of soluble protein in seeds that appeared empty on radiographs. By comparison, unfilled (aborted) seeds contained significantly less insoluble and soluble protein than empty seeds that were fed-upon by L. occidentalis; moreover, no crystalloid (11S globulin) breakdown products were generated. The biochemical markers described in this study are reliable tools that can be used to identify conifer seeds that have sustained light to severe damage from L. occidentalis feeding.

Changes in volatile terpene and diterpene resin acid composition of resistant and susceptible white spruce leaders exposed to simulated white pine weevil damage.

Induced (traumatic) resin in white spruce (Picea glauca (Moench) Voss) leaders resistant or susceptible to the white pine weevil (Pissodes strobi Peck) was analyzed for volatile terpenes and diterpene resin acids after simulated white pine weevil damage. Leaders from 331 trees were wounded just below the apical bud with a 1-mm diameter drill, coinciding with the natural time of weevil oviposition in the spring. Leaders were removed in the fall, and the bark and xylem from the upper and lower regions of the leader extracted and analyzed by gas chromatography. Unwounded trees had low amounts of resin in xylem compared with bark. In response to wounding, volatile terpenes and diterpene resin acids increased in the upper xylem (area of wounding), with resistant trees showing a greater increase than susceptible trees. Wounding caused monoterpenes in particular to decrease in the lower region of the leader (away from the drilled area) in greater amounts in susceptible trees than in resistant trees. In response to wounding, the proportion of monoterpene to resin acid increased in the upper and lower xylem of resistant trees, and slightly increased in the upper xylem of susceptible trees. Monoterpene-enriched resin is more fluid than constitutive resin, and probably flows more readily into oviposition cavities and larval mines, where it may kill immature weevils. Loss of resin components in the lower xylem suggested catabolism and transport of these materials to the site of wounding; however, energetic and regulatory data are necessary to confirm this hypothesis. This study provides a basis for measuring the ability of a tree to undergo traumatic resinosis that could be used to screen for resistance to white pine weevil.

Effect of feeding by the western conifer seed bug, Leptoglossus occidentalis, on the major storage reserves of developing seeds and on seedling vigor of Douglas-fir.

The impact of feeding by the western conifer seed bug (Leptoglossus occidentalis Heidemann) on storage reserves of developing seeds of Douglas-fir (Pseudostuga menziesii (Mirb.) Franco) was studied by caging nymphal and adult seed bugs on cones during late-season development, and nymphs on cones during early, mid- and late-season development. Analysis of the major storage reserves of partially damaged seeds revealed that late-season feeding by each life stage and feeding by nymphs at all three stages of cone development significantly reduced the amounts of lipid and buffer-insoluble (crystalloid) protein in seeds at harvest by up to 78 and 97%, respectively. Seeds showing light to moderate damage on radiographs did not exhibit a reduction in the amount of buffer-soluble (matrix) protein. Seeds damaged by feeding during early development compensated in part by continuing to synthesize lipid and crystalloid protein. Light or moderate damage to mature Douglas-fir seeds exposed to L. occidentalis in the laboratory reduced seedling emergence by > 80%, but the seedlings that emerged successfully appeared to suffer no adverse effects when grown under standard nursery conditions.

Impact of Leptoglossus occidentalis (Hemiptera: Coreidae) on Douglas-fir seed production.

We investigated the effect of feeding by the western conifer-seed bug, Leptoglossus occidentalis Heidemann, on seed production in developing cones of coastal Douglas-fir, Pseudostuga menziesii (Mirb.) Franco, with respect to seed bug life stage and sex (nymphs, adult females, and adult males) and timing of feeding (early, mid-, and late season cone development). Feeding by females on caged cones for a 2-wk period during late season cone development reduced the proportion of full seeds in cones by approximately 70% compared with caged control cones. There was no significant difference among nymphs, adult females, and adult males with respect to the proportion of empty or partially fed-upon seeds produced during the same feeding period. Feeding by nymphs for 2 wk early in the season resulted in a threefold increase in the number of unextractable seeds fused to cones compared with the control. Weight measurements of harvested seeds indicated that radiography is an accurate tool to distinguish among Douglas-fir seeds that have sustained light, moderate, or severe damage. Determining the full impact of L. occidentalis on conifer seed production will require the development of a reliable method to distinguish between naturally aborted seeds and seeds emptied through feeding by seed bugs.

Response of mountain pine beetle,Dendroctonus ponderosae Hopkins, and pine engraver,Ips pint (SAY), to ipsdienol in southwestern British Columbia.

In bioassays conducted with walking beetles in the laboratory (S)-(+)-, (R)-(-)-, and (±)-ipsdienol were attractive alone, but reduced the attraction of both sexes of the mountain pine beetle,Dendroctonus ponderosae Hopkins, to extracts of female frass. Field trapping studies indicated that attraction ofD. ponderosae to a stimulus composed of myrcene +trans-verbenol +exo-brevicomin was significantly reduced with the addition of (±)- and sometimes (S)-(±)-ipsdienol. Thus, (S)-(+)-ipsdienol produced by males of this species may act as an antiaggregation pheromone. (S)-(+)-Ips-dienol is thought to function as a repellent allomone against the pine engraver,Ips pini (Say), in regions whereI. pini utilizes (R)-(-)-ipsdienol as an aggregation pheromone. However, in southwestern British ColumbiaI. pini was attracted to the (±)-ipsdienol used in field bioassays ofD. ponderosae, a finding consistent with the production of both enantiomers byI. pini in this region. When presented with the ternary semiochemical bait forD. ponderosae, (±)-ipsdienol was not attractive toI. pini. Thus, the activity of (S)-(+)-ipsdienol as a repellent allomone againstI. pini seems to be replaced in southwestern British Columbia by the inhibitory effects of myrcene,trans-verbenol,exo-brevicomin, or some combination thereof.

Olfactory discrimination byHeteropsylla cubana (Homoptera: Psyllidae) between susceptible and resistant species ofLeucaena (Leguminosae).

In the field, adult psyllids,Heteropsylla cubana Crawford, oriented significantly more towards the caged seedlings of susceptibleLeucaena leucocephala (Lam.) de Wit than to those of the resistant tree species,Leucaena collinsii Britton & Rose, or the nonhostAmaranthus spinosus L. In a dual-choice bioassay using a still-air olfactometer, the females demonstrated a strong positive response to the hexane extract ofL. leucocephala leaves at 1×10(-3) g equivalents (g eq) of leaf material. The females did not orient to leaf extracts ofL. collinsii at high concentrations but responded positively at a lower concentration of 1×10(-4) g eq. Olfactory discrimination byH. cubana between resistant and susceptible host species should be considered in selection and breeding programs.

Terpene alcohol pheromone production byDendroctonus ponderosae andIps paraconfusus (Coleoptera: Scolytidae) in the absence of readily culturable microorganisms.

Dendroctonus ponderosae Hopkins andIps paraconfusus Lanier of both sexes produced most of their complement of terpene alcohols at normal to elevated levels in the absence of readily culturable microorganisms. However, there was some evidence that microbial involvement may be required by maleI. paraconfusus to produce ipsenol and ipsdienol at normal levels. Increased levels of certain terpene alcohols found in axenically reared or streptomycin-fed beetles suggest that symbiotic microorganisms may be responsible for breaking down pheromones and other terpene alcohols. There was also evidence for microbial involvement in the production of the antiaggregation pheromone verbenone inD. ponderosae. This compound was not produced in quantifiable levels by axenically reared or streptomycin-fed beetles exposed to α-pinene as vapors or through feeding, but was found in wildD. ponderosae exposed to α-pinene through feeding on bolts of lodgepole pine,Pinus contorta var.latifolia Engelmann.

Conversion of verbenols to verbenone by yeasts isolated fromDendroctonus ponderosae (Coleoptera: Scolytidae).

A variety of symbionts associated with bark beetles are capable of producing compounds that are used as pheromones by their hosts. We report that two yeasts associated withDendroctonus ponderosae Hopkins,Hansenula capsulata Wickerham, andPichiapinus (Hoist) Phaff, are capable of convertingcis- andtrans-verbenol efficiently into verbenone.trans-Verbenol, which is produced by femaleD. ponderosae, acts as an aggregation pheromone for this scolytid, while verbenone, which other studies have indicated that microbe-reducedD. ponderosae are incapable of producing, acts as an antiaggregation pheromone.D. ponderosae appears to rely primarily on microbial symbionts for terminating aggregation and mass attack on individual host trees.

ß-Phellandrene: kairomone for pine engraver,Ips pini (Say) (Coleoptera: Scolytidae).

The responses ofIps pini (Scolytidae) to multiple-funnel traps baited with the pheromone, ipsdienol, and various monoterpenes were determined in stands of lodgepole pine in southern and central British Columbia.Ips pini was attracted to both ipsdienol and ß-phellandrene, demonstrating that ß-phellandrene is a kairomone for this species.Lasconotus complex (Colydiidae) and aCorticeus sp. (Tenebrionidae) were attracted to both ipsdienol and ß-phellandrene. TheCorticeus sp. exhibited a synergistic response to the combination of ipsdienol and ß-phellandrene; the responses of the other two species to the combination were additive. The predators,Thanasimus undatulus, Enoclerus sphegeus, andE. lecontei (Cleridae), were attracted to ipsdienol-baited traps, whileMonochamus clamator (Cerambycidae) andDendroctonus ponderosae (Scolytidae) were attracted to ß-phellandrene. Attraction of all eight species increased with increasing release rates of ipsdienol and/or ß-phellandrene.

Influence of different habitats and mating on olfactory behavior of onion flies seeking ovipositional hosts.

Using traps baited with natural and synthetic onion volatiles, we examined the effects of different habitats and mating on the olfactory behavior of laboratory-reared and wild onion flies. Rankings of olfactory treatments as host-finding stimuli for females were dependent on their mating status and the habitat in which they were foraging. In habitats devoid of hosts, traps baited with individual alkyl sulfides were as effective as 4-day-old chopped onions and more effective than 1-day-old onions in eliciting host-finding behavior in laboratory-reared unmated females (LUF) and laboratory-reared mated females (LMF). However, upwind dispersal and percent recapture were always significantly greater in LUF. In one experiment, Pr2S2 was 19 times more attractive to LMF in a fallow field, as than it was in an onion field. Reduced effectiveness of alkyl sulfides as host-finding stimuli in onion fields probably results in part because they are less findable, but more importantly because of a change in searching behavior after females have mated. Evidence to support the latter contention is that traps baited with alkyl sulfides and onions were equally findable by unmated females in both habitats. The behavior of LMF was identical to that of wild females, whereas the behavior of LUF was identical to wild males. The hypothesis that olfactory host-finding behavior in onion flies is modified by the resource level was upheld. Alkyl sulfides appear to be the primary, and possibly the only, chemical effectors of host-finding at the patch level of resource distribution, whereas the complex blend emitted by aged, chopped, or damaged onions appears to be acting at the final level of host-finding, while egg-laying females are moving between adjacent hosts in search of an optimal oviposition site.

Aggregated oviposition inDelia antiqua (Meigen): A case for mediation by semiochemicals.

Experiments conducted in the laboratory tested the hypotheses that aggregated oviposition by onion maggot flies,Delia antiqua (Meigen), is caused by stimuli associated with ovipositing females, newly laid eggs, or both. Using a paired oviposition station bioassay that eliminated visual stimuli associated with the treatment under study, 67% of the eggs laid by caged females were in response to the odor of females already ovipositing on an onion slice, as opposed to 33% of the eggs laid in response to an onion slice alone. When newly laid eggs were transferred to onion slices and held for either 24 or 48 hr before being bioassayed against similarly aged untreated onions, 74% and 97% of the eggs were laid at the egg-treated onion stations, respectively. Similar results were achieved when an aqueous wash of newly laid eggs was applied to the onion slice. When the egg wash was processed through a bacterial filter or when eggs were present but not in contact with onions, all response was eliminated. These results implicate microorganisms transmitted on the egg surface in creating an attraction for ovipositing females. Heptane extracts of ovipositor tips from mated, ovipositing females induced 72% of the test females to oviposit near points at which extracts were applied to the oviposition station floor. A behavioral sequence for an optimal host-selection strategy is hypothesized, whereby host-seeking female onion flies respond to host-derived alkyl sulfides at long range and metabolic by-products of microbially infested hosts and visual cues at short range (ca. l m), with final selection of oviposition sites potentially reinforced by contact with an aggregation pheromone released or left on the substrate by ovipositing females.

(S)-(+)-Ipsdienol: Interspecific inhibition ofIps latidens (leconte) byIps pini (Say) (Coleoptera: Scolytidae).

In south-central British Columbia, the attraction ofIps latidens (LeConte) to its pheromone, ipsenol, was inhibited by (S)-(+)-ipsdienol, a pheromone forI. pini (Say). (R)-(-)-lpsdienol had no effect onI. latidens. (S)-(+)-lpsdienol probably plays a role in interspecific communication between the two species, facilitating reductions in interspecific competition for breeding material and/or interspecific mating interference.

Repellency of oral exudate to eastern and western spruce budworm larvae (Lepidoptera: Tortricidae).

A two-choice feeding bioassay was used to investigate the intra-specific repellency of the larval oral exudate of eastern and western spruce budworms,Choristoneura fumiferana (Clem.) andC. occidentalis Free., respectively. Results of the bioassay indicated that feeding behavior on artificial diet-drop feeding stations was modified in the presence of exudate, with feeding stations treated with conspecific exudate being avoided when an untreated station was available 3 cm away. Feeding was suppressed when a single, exudate-treated station was provided, or when the treated and untreated stations were separated by only 1 cm. The repellent effect functioned both inter- and intraspecifically. When induced to produce exudate,C. occidentalis larvae were not immediately repelled by either their own or other individuals' exudate. However, 24 hr after induction, test larvae were repelled by exudate from either source. In both species, larval oral exudate probably functions to repel conspecific competitors.

Angiosperm bark volatiles disrupt response of Douglas-fir beetle, Dendroctonus pseudotsugae, to attractant-baited traps.

Antennally active, bark-derived, angiosperm volatiles were tested singly and in groups for their ability to disrupt the response of the Douglas-fir beetle (DFB), Dendroctonus pseudotsugae, to attractant-baited multiple-funnel traps. One compound, conophthorin, was active alone in reducing the response of beetles to the baited traps. Further experiments showed disruptive activity in two aliphatic green-leaf alcohols [1-hexanol and (Z)-3-hexen-1-ol], as well as guaiacol and benzyl alcohol, and three aliphatic aldehydes [nonanal, hexanal, and (E)-2-hexenal] but not in two aromatic aldehydes (benzaldehyde and salicylaldehyde). Every binary combination that included conophthorin or any two of the other groups, except aromatic aldehydes, significantly reduced the response of beetles to baited traps. Various ternary mixtures and the complete mixture of all the groups were generally the most effective treatments. These results provide evidence that DFBs recognize and avoid nonhosts while flying rather than landing on candidate hosts and testing them while in contact with the tree. Nonhost angiosperm bark volatiles may have practical utility on their own or in combination with the antiaggregation pheromone 3-methylcyclohex-3-en-1-one (MCH) to protect single trees, logs, or stands from attack by the DFB.

Sex pheromone of the mullein bug,Campylomma verbasci (Meyer) (Heteroptera: Miridae).

The first known sex pheromone for the family Miridae is reported for the mullein bug,Campylomma verbasci (Meyer) (Heteroptera: Miridae). Extracts of and Porapak Q-trapped volatiles from females were attractive to males in field bioassays. Butyl butyrate and hexyl butyrate were the predominant compounds in the females' volatiles, but these compounds previously had been found to be inactive when released alone or in combination. Butyl butyrate in combination with an isolated trace chemical, (E)-crotyl butyrate (which was inactive alone), rivaled the attraction elicited by five live females. When synthetic butyl butyrate and (E)-crotyl butyrate in a 16: 1 ratio were released at 91 or 183 µg/day, they comprised an effective attractant for male bugs.