Field bioassay ofexo- andendo-brevicomin withDendroctonus ponderosae in lodgepole pine

Bothexo- andendo-brevicomin reduced the response of flyingDendroctonus ponderosae (Coleoptera: Scolytidae) to an attractant composed oftrans-verbenol and terpenes or oftrans- andcis-verbenol and terpenes in a stand ofPinus contorta var.murrayana. These data suggest that racemicexo- andendo-brevicomin may interrupt aggregation in populations of mountain pine beetle colonizing lodgepole pine; functions of the natural chiral compounds are unknown.Research supported by NSF grants DEB 77-11367 and PCN 79-21708 and the O.S.U. School of Forestry. O.S.U. Agric. Exp. Sta. Tech. Paper No. 5713.     

Attractive and inhibitory pheromones produced in the bark beetle,Dendroctonus brevicomis,during host colonization: Regulation of inter- and intraspecific competition

Quantities of attractive (exo-brevicomin and frontalin) and inhibitory (trans-verbenol, verbenone, and ipsdienol) pheromones were monitored in both sexes ofDendroctonus brevicomis during their colonization of a ponderosa pine. Verbenone was found in males in the greatest amounts at the time of landing, and it declined more rapidly than the other pheromones in either sex. The amounts of frontalin andexo-brevicomin in males and females, respectively, increased after initial boring within the host but began to decline after mating. The quantity oftrans-verbenol in both sexes (females had significantly more) declined more gradually thanexo-brevicomin, frontalin, and verbenone. Ipsdienol was found only in males during the initial stages of attack when encountering the resin. It is suggested that along with a general decline in all pheromonal components, a sufficient change in the ratio of the attractive pheromones to an inhibitory pheromone,trans-verbenol, may play a role in termination of aggregation.trans-Verbenol may also function along with verbenone and ipsdienol in limiting the density of attack and thus intraspecific competition. These inhibitory pheromones also appear to cause several competing species of bark beetle to avoid landing in areas infested withD. brevicomis, even when their own pheromone is present.Coleoptera:Scolytidae. These studies were supported in part by the Rockefeller Foundation, USDA Forest Service, Regional research project W-110, SEA/ USDA, and the National Science Foundation and Environmental Protection Agency through a grant (NSF GB-34719/BMS 75-04223) to the University of California.     

Field responses of the western pine beetle1 and one of its predators to host- and beetle-produced compounds

The responses of the western pine beetle (Dendroctonus brev-icomis LeConte) andTemnochila chlorodia (Mannerheim) to candidate attractants—exo- andendo-brevicomm, frontalin,trans-verbenol, ver-benone, and ponderosa pine turpentine and its major monoterpene components—were quantified by counts of beetles on traps baited with the various attractants, singly and in combinations released simultaneously. Combinations ofexo-brevicomin and frontalin plus a monoterpene or turpentine were the most attractive toD. brevicomis. The responses to these attractant combinations were reduced when verbenone plustrans-verbenol were present. All single compounds and binary mixtures, exceptexo-brevicomin plus frontalin, were much less attractive.exo-Brevicomin was most attractive toT. chlorodia, and this response appeared to decrease when verbenone plustrans-verbenol were present.Coleoptera: Scolytidae.Coleoptera: Trogositidae.This research was supported in part by the Forest Service, U.S. Department of Agriculture, and in part by grants from the Rockefeller Foundation to the University of California. Commercial enterprises and products are mentioned solely for information, and do not imply endorsement by the sponsoring agencies and organizations.     

Interspecific inhibition of the response of the bark beetles,Dendroctonus brevicomis andIps paraconfusus,to their pheromones in the field

Ponderosa pine logs infested withIps paraconfusus males inhibited the attraction ofDendroctonus brevicomis in the field to either attractive logs cut from a ponderosa pine tree under attack byD. brevicomis or to their synthetic pheromones,exo-brevicomin, frontalin, and myrcene. Logs cut from trees under attack byD. brevicomis inhibited the response ofI. paraconfusus to logs infested with maleI. paraconfusus.Exo-brevicomin, frontalin, and myrcene did not inhibit their response but verbenone did. Verbenone was found in maleD. brevicomis dissected from attractive logs under attack during the same time the response ofI. paraconfusus was inhibited by these logs.Trans-verbenol andexo-brevicomin were found in femaleD. brevicomis while verbenone,trans-verbenol, and frontalin were found in maleD. brevicomis in relatively large amounts near the beginning of the aggregation phase of host colonization. All of these compounds had decreased at a similar rate 5 days later. This gradual decrease inexo-brevicomin and frontalin probably caused the observed reduction in attraction. The ecological significance of these compounds in relation to termination of the aggregation phase ofD. brevicomis and reduction of interspecific competition is discussed.Coleoptera: Scolytidae. This paper is based in part upon a dissertation submitted by J.A.B. to the University of California, Berkeley, in partial fulfillment of the requirements for the PhD in entomology. These studies were supported in part by the Rockefeller Foundation; U.S. Forest Service; Regional research Project W-110, S.E.A./ U.S.D.A.; and the National Science Foundation and Environmental Protection Agency through a grant (NSF GB-34719/BMS 75-04223) to the University of California.     

Electroantennogram responses by mountain pine beetles,Dendroctonus ponderosae Hopkins,exposed to selected semiochemicals

Electroantennograms (EAGs) were obtained forD. ponderosae to the bark beetle pheromonestrans-verbenol,cis-verbenol,exo-brevicomin,endo-brevicomin, frontalin, verbenone; to the kairomones, -pinene, -pinene, camphene, -3-carene, limonene, myrcene; to a blend (111) oftrans-verbenol,exo-brevicomin, and myrcene; and to diacetone-alcohol. Male and female responses, in general, did not differ significantly over the whole EAG dose-response curves but differed at a few concentrations on many of the curves. There were more differences noted for pheromones than kairomones. The blend yielded among the largest EAGs in both sexes and appeared to show synergism. Responses of females were lower than those for males in most instances. Significant differences in responses by the two sexes were much fewer for the kairomones than the pheromones. EAG recovery rates tested at only one concentration showed significant differences between males and females for three pheromones,trans-verbenol,cis-verbenol, and verbenone, and two kairomones, camphene and -3-carene. Thresholds were quite low for most of the odorants exceptcis-verbenol, camphene, verbenone, and diacetone-alcohol in females, andcis-verbenol, verbenone, -pinene, and diacetone-alcohol in males. The results, using at least one EAG parameter, support behavioral and field studies involvingexo-brevicomin,trans-verbenol, frontalin, and the blend.     

Electroantennograms by mountain pine beetles,Dendroctonus ponderosae Hopkins,exposed to selected chiral semiochemicals

Electroantennograms (EAGs) were recorded fromD. ponderosae to the enantiomers of the terpenoid bark-beetle pheromonestrans-verbenol,cis-verbenol, verbenone, and the bicyclic ketals frontalin,exo-brevicomin, andendo-brevicomin. Male and female responses to enantiomers of the terpenoids differed significantly only at the two highest concentrations. No sex differences were seen in response to the bicyclic ketals. Significantly different responses to the enantiomers of all the chemicals, except frontalin, were noted over at least part of the dosage-response ranges tested. The negative antipode for all of the terpenoids elicited higher responses, while for the bicyclic ketals, the positive antipode effected the largest responses except for the two highest concentrations ofexo-brevicomin.     

Olfactory receptor cell responses ofIps grandicollis (eichhoff) (Coleoptera: Scolytidae) to intra- and interspecific behavioral chemicals

Electrophysiological recordings from antennal olfactory receptor cells were obtained fromIps grandicollis. Recordings were made from olfactory receptor cells from nine regions of the antennae in response to stimulation with the semiochemicals-pinene, frontalin,endo-brevicomin, verbenone,trans-verbenol,cis-verbenol, ipsdienol, and ipsenol. In many cases, up to two cells were recorded concurrently from the same location. When compared to males, females had a greater percentage of cells responsive to the primary pheromones ofDendroctonus frontalis, frontalin andtrans-verbenol, and ofIps spp., ipsdienol and ipsenol. Among females, more cells responded totrans-verbenol and theIps-produced volatiles than to host or otherD. frontalis-produced compounds. Olfactory cells of males responded mostly tocis-verbenol, followed by-pinene, verbenone,trans-verbenol, andendo-brevicomin. Of those cells responsive primarily to one compound, the greatest percentage were responsive totrans-verbenol in females and to verbenone in males. The response of the antennal olfactory receptor cells to semiochemicals used by male and femaleI. grandicollis is consistent with the presence of these compounds during the host colonization period for each sex. Our results, which show a lack of specificity in most pheromone and host odor receptor cells, is in contrast with previously published accounts of olfactory receptor cell specificity in otherIps species.     

Male-specific conversion of the host plant compound,myrcene, to the pheromone, (+)-ipsdienol,in the bark beetle,Dendroctonus brevicomis

When both sexes of the bark beetle,Dendroctonus brevicomis LeConte, were exposed to vapors of myrcene from ponderosa pine, only the male produced (+)-ipsdienol. In the field, racemic ipsdienol significantly reduced the attraction of both sexes in flight to a mixture of myrcene and the aggregation pheromones,exo-brevicomin and frontalin. This suggests that ipsdienol may be involved in regulating colonization density ofD. brevicomis. The implications of the biosynthesis of various enantiomers of ipsdienol byD. brevicomis and the cohabitating bark beetles,Ips paraconfusus andI. pini, in relation to their behavioral responses are discussed in regard to reducing interspecific competition.Coleoptera: Scolytidae.     

Secondary attraction and field activity of beetle-produced volatiles inDendroctonus terebrans

Controlled infestation ofDendroctonus terebrans (Olivier) on bolts of slash pine,Pinus ellionii Engelm. var.elliottii, elicited greater attraction of wild conspecifics than uninfested bolts. Secondary attraction was not apparent, however, to standing slash pines that had received volunteer attacks, when compared with attraction to unattacked but susceptible trees. Hindguts from in-flight or attacking femaleD. terebrans contained frontalin, and those from in-flight or attacking males containedexo-brevicomin.Trans-pinocarveol,cis-verbenol,trans-verbenol, myrtenal, verbenone, myrtenol, and other compounds were produced by both sexes during gallery construction in host trees. Synthetic frontalin, when deployed with a standard host odor mixture of turpentine and ethanol, was very attractive to maleD. terebrans in field-trapping experiments. The addition of eitherexo-brevicomin orendo-brevicomin to the frontalin-turpentine combination negated the attractive effect of frontalin for males.Trans-verbenol, myrtenol, and verbenone had little effect onD. terebrans behavior. Responses of females did not differ among treatments in any of the 11 field experiments.     

Monoterpene metabolism in female mountain pine beetles,Dendroctonus ponderosae Hopkins,attacking ponderosa pine

Abdominal volatiles of female mountain pine beetles,Dendroctonus ponderosae Hopkins, fed in ponderosa pine,Pinus ponderosa Dougl. ex Laws, and in lodgepole pine,P. contorta var.latifolia Engelmann, were analyzed by gas chromatography and coupled gas chromatography-mass spectrometry and were found to comprise host oleoresin components and beetle-produced alliylic alcohols, aldehydes, and ketones derived from host monoterpenes. Neitherexo- andendo-brevicomin nor frontalin were detected. Three metabolic pathways are proposed to account for the distribution of beetle-produced monoterpene alcohols. The first pathway involves hydroxylation of monoterpene substrates on allylic methyl groups which areE to a methylene or vinyl group. This oxidation pathway is indiscriminate with respect to substrate and probably functions to detoxify monoterpenes. A second pathway, which hydroxylates theendo-cyclic methyleneE to a vinyl methyl group of bicyclic monoterpenes to give almost exclusively thetrans alcohol, is hypothesized to be involved in pheromone production. A third detoxification pathway involves anti-Markovnikov addition of water to theexo-cyclic double bond of -phellandrene to give predominantlytrans-2-p-menthen-7-ol.Coleoptera: Scolytidae.Research supported by the Natural Sciences and Engineering Research Council of Canada: Operating Grant A3881 and A3706; Strategic Grant G1039; and Science Council of British Columbia Grant 33 (RC-4).     

Semiochemicals produced by western balsam bark beetle,Dryocoetes confusus Swaine (Coleoptera: Scolytidae)

The most prominent beetle-produced volatiles identified in the abdominal extracts of maleDryocoetes confusus Swaine after they had bored for 24 hr in logs of subalpine fir,Abies lasiocarpa (Hook.) Nutt. were:exo andendo-brevicomin,trans-verbenol, verbenone, myrtenol,trans-pinocarveol,cis- andtrans-p-menthen-7-ol, 3-caren-10-ol, and several monoterpenes and sesquiterpenes. Myrtenol was the only conspicuous compound in extracts from males that had been exposed toA.lasiocarpa resin volatiles for 24 hr. Laboratory bioassays indicated that both (–)- and (+)-exo-brevicomin were attractive to femaleD. confusus, and that the (–) enantiomer did not inhibit response to its antipode. Results from field trapping experiments indicated that bothero-brevicomin and myrtenol are aggregation pheromones forD. confusus.exo-Brevicomin baits were effective in causing attack byD. confusus on baited and surrounding trees, suggesting that this pheromone may have utility in manipulating populations of the beetle.Research supported by the Natural Sciences and Engineering Research Council of Canada, Operating Grants A3881 and A3706 and Strategic Grant G1039.     

(E)- and (Z)-6-nonen-2-one: Biosynthetic precursors ofEndo- andexo-brevicomin in two bark beetles (Coleoptera: Scolytidae)

The ability of (E)- and (Z)-6-nonen-2-one to serve as precursors of the common scolytid pheromonesEndo- andexo-brevicomin was examined in vivo. When mountain pine beetles (MPB),Dendroctonus ponderosae Hopkins, or western balsam bark beetles (WBBB),Dryocoetes confusus Swaine, were exposed to [6,7-D₂](E)-6-nonen-2-one, theEndo-brevicomin produced was enriched with two deuterium atoms per molecule (as determined by GC-MS), indicating that (E)-6-nonen-2-one served as a precursor of this pheromone. Similarly, when the beetles were exposed to [4,4-D₂](Z)-6-nonen-2-one, theexo-brevicomin produced was enriched with two deuterium atoms per molecule. Evidence in support of biological relevance of the latter observation include: (1) (Z)-6-nonen-2-one was found in the volatiles of male MPBs and WBBBs, indicating that this is a natural metabolite; (2) theexo-brevicomin produced by MPB was shown to be of natural (+) chirality by complexation chromatography; and (3) female WBBBs and MPBs (which are not known to produceexo-brevicomin) produced significantly lessexo-brevicomin when exposed to the precursor than did the males.     

Pheromone production by axenically rearedDendroctonus ponderosae andIps paraconfusus (Coleoptera: Scolytidae)

Mountain pine beetles,Dendroctonus ponderosae Hopkins, and California five-spined ips,Ips paraconfusus Lanier, were reared axenically from surface-sterilized eggs on aseptic pine phloem. After 24 hr in host logs, axenip femaleD. ponderosae and maleI. paraconfusus produced the aggregation pheromones,trans-verbenol (D. ponderosae), and ipsenol and ipsdienol (I. paraconfusus). Emergent, axenically reared maleD. ponderosae contained normal amounts of the pheromoneexo-brevicomin. Axenic femaleD. ponderosae treated with juvenile hormone or exposed to vapors of -pinene, produced the pheromonetrans-verbenol. By 25–35 days after eclosion, axenic females exposed to -pinene vapors produced over six times as muchtrans-verbenol as wild females, suggesting that while microorganisms in wild females may producetrans-verbenol, they may also inhibit production of the pheromone or use it as a substrate.Research supported in part by Natural Sciences and Engineering Research Council, Canada, Operating Grants A3881 and A3706, and H.R. MacMillan Family Fund Fellowships to J.E. Conn and D.W.A. Hunt.     

The response of mountain pine beetle (Dendroctonus ponderosae) to lodgepole pine trees baited with verbenone and exo-brevicomin

exo-brevicomin, a multifunctional pheromone of the mountain pine beetle,Dendroctonus ponderosae, was tested at release rates of 0.5 and 2.5 mg/day alone and in combination with the antiaggregation pheromone verbenone against unbaited controls. Significantly more lodgepole pinePinus contorta var.latifolia trees were attacked, and at higher densities, with both release rates ofexo-brevicomin than with all other treatments. Verbenone significantly reduced the response of mountain pine beetles toexo-brevicomin. Verbenone alone did not reduce the number of trees attacked by mountain pine beetle or the attack density when compared to the unbaited controls.     

Effects of verbenone andtrans-verbenol on the response ofDendroctonus brevicomis to natural and synthetic attractant in the field

Verbenone andtrans-verbenol were investigated as candidate interruptants for use as tree protectants. Verbenone andtrans-verbenol, pheromones released byDendroctonus brevicomis during host colonization, reduced the trap catch ofD. brevicomis near sources of the attractant composed ofexo-brevicomm, frontalin, and myrcene. Catch reduction at some trap positions was greater at a high release rate than at a low release rate oftrans-verbenol alone and of the combination of verbenone plustrans-verbenol. Verbenone also reduced catches at traps baited with attractive bolts from trees under attack byD. brevicomis. Attempts to use verbenone to protect living trees fromD. brevicomis attack were inconclusive.Coleoptera: Scolytidae.This research was supported in part by the Forest Service, U.S. Department of Agriculture, and in part by grants from the National Science Foundation/Environmental Protection Agency (grant NSF-6B-34718/BMS75-04223), and in part by grants from the Rockefeller Foundation to the University of California. Mention of a proprietary or commercial product does not constitute recommendation or endorsement of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products that also may may be suitable.     

Interruption of response ofDendroctonus brevicomis to its attractive pheromone by components of the pheromone

The catch of the western pine beetle,Dendroctonus brevicomis, at an attractive source of racemicexo-brevicomin, racemic frontalin, and myrcene was reduced by surrounding the source with a grid of 48 stations releasing all three compounds together, orexo-brevicomin alone or myrcene alone. Each compound was released at the rate of 2 mg/24 hr/station. The catch at an attractive bolt cut from a tree being colonized byD. brevicomis was not reduced byexo-brevicomin, but was reduced by the combination ofexo-brevicomin, frontalin, and myrcene in one of two tests. When a transect of traps was placed across a 0.81-hectare plot at six of the 48 stations releasing all three compounds, more beetles were caught at outer than at inner traps. More beetles were caught at unbaited traps on trees in a plot when the three compounds were released than when onlyexo-brevicomin or no compounds were released. A few trees were attacked byD. brevicomis in some of the plots. The antiattractant verbenone released from 48 stations at the rate of 4 mg/24 hr/station did not reduce the catch at an attractive tree bolt.Coleoptera: Scolytidae.These studies were supported by the Forest Service, U.S. Department of Agriculture, and in part by grants to the University of California from the U.S. National Science Foundation and U.S. Environmental Protection Agency (NSF GB-34718/BMS 75-04223) and the Rockefeller Foundation. The findings, opinions, and recommendations are not necessarily those of the funding agencies. Mention of commercial enterprises or products is only for information.     

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.Research supported in part by Natural Sciences and Engineering Research Council, Canada, Operating Grant A3881, Strategic Grant G1039, a Postgraduate Scholarship to D.W.A. Hunt, and the Science Council of British Columbia Grant 1 (RC-10)     

Identification of Semiochemicals Associated with Jeffrey Pine Beetle, Dendroctonus jeffreyi

Analysis of frass extracts and aerations from adult beetles confirmed earlier studies identifying 1-heptanol as a principal pheromone constituent produced by female Dendroctonus jeffreyi Hopkins. The aerations also indicated that males produced 33% (+) and 67% (–)-frontalin, previously unreported for this species, and >99% (+)-exo-brevicomin. Both males and females were responsive to these compounds in electrophysiological studies. Laboratory bioassays and field trapping experiments indicated that males and females were attracted to a mixture of 1-heptanol plus the principal host resin component heptane, but females were less attracted than males. Inclusion of racemic exo-brevicomin at 0.1% concentration in the traps increased the response of females. Inclusion of racemic frontalin above a 1% concentration in the normally attractive lures blocked arrival of beetles to the traps. While the female-produced pheromone components and host synergists are very different between the sibling species D. jeffreyi and Dendroctonus ponderosae Hopkins, the principal male-produced compounds are the same, and the response of the female beetles to those compounds appears to be similar.     

Sensitivity of antennae of male and femaleIps paraconfusus (Coleoptera: Scolytidae) to its pheromone and other behavior-modifying chemicals

The antennal sensitivities of both male and femaleIps paraconfusus were found generally to be greatest for conspecific aggregation pheromones (ipsdienol, ipsenol); intermediate for an additional conspecific pheromone (cis-verbenol), an aggregation synergist (2-phenylethanol), and pheromones/allomones of sympatric species (trans-verbenol, verbenone, and frontalin); and lowest for both host terpenes (alpha-pinene and myrcene) and other bark beetle-produced odorants (exo-brevicomin and linalool). Of the enantiomeric compounds tested, antennae of both sexes did not differ in sensitivity between thetrans-verbenol enantiomers at low dosage levels; but at higher dosages, the conspecific-produced enantiomer, (1R,4S,5R)-(+)-trans-verbenol, elicited larger mean EAG responses than its antipode, (1S,4R, 5S)-(?)-trans-verbenol. At the mid-dosage range, female antennae tended to be slightly more responsive to (S)-(?)-verbenone than to (R)-(+)-verbenone, while male antennae were equally responsive to stimulations by either verbenone enantiomer. In field bioassays there was a large and significant reduction in trap catches ofI. paraconfusus on traps where the (S)-(?)- or (R)-(+)-enantiomers of verbenone were evaporated beside logs containing boring conspecific males. Only when the (S)-(?)-enantiomer of verbenone was evaporated beside logs containing boring males did the sex ratio ofI. paraconfusus trapped shift from female-dominated to male-dominated attraction. Thus both physiological and behavioral data suggest a differential chiral sensitivity of female beetles for the verbenone enantiomers. The relative sensitivities between different chiral compounds derived from one or the other of the common precursoral host terpenes, (S)-(?)- and (R)-(+)alpha-pinene or myrcene, are discussed.     

Ipsenol: an aggregation pheromone forIps latidens (Leconte) (Coleoptera: Scolytidae)

Ipsenol was identified from the frass of male, but not female,Ips latidens from British Columbia, feeding in phloem tissue of lodgepole pine,Pinus contorta var.latifolia. The responses ofJ. latidens to sources of ips-enol andcis-verbenol were determined with multiple-funnel traps in stands of lodgepole pine in British Columbia. Ipsenol attracted both male and femaleI. latidens, verifying that it is a pheromone for this species. MaleI. latidens showed a slight preference for (S)-(–)-ipsenol.cis-Verbenol was not produced by beetles of either sex and, in contrast to an earlier report, both enantiomers inhibited attraction to ipsenol-baited traps. The predators,Enoclerus sphegeus andThanasimus undatulus (Cleridae), were attracted to traps baited withcis-verbenol and ipsenol.