Acquired and Partially De Novo Synthesized Pyrrolizidine Alkaloids in Two Polyphagous Arctiids and the Alkaloid Profiles of Their Larval Food-Plants

The profiles of pyrrolizidine alkaloids (PAs) in the two highly polyphagous arctiids Estigmene acrea and Grammia geneura and their potential PA sources in southeastern Arizona were compiled. One of four species of Boraginaceae, Plagiobothrys arizonicus, contained PAs; this is the first PA record for this plant species. The principle PA sources are Senecio longilobus (Asteraceae) and Crotalaria pumila (Fabaceae). The known PA pattern of S. longilobus was extended; the species was found to contain six closely related PAs of the senecionine type. Three novel PAs of the monocrotaline type, named pumilines A-C, were isolated and characterized from C. pumila, a species not studied before. The pumilines are the major PAs in the seeds, while in the vegetative organs they are accompanied by the simple necine derivatives supinidine and as the dominant compound subulacine (1beta,2beta-epoxytrachelanthamidine). In both plant species, the PAs are stored as N-oxides, except C. pumila seeds, which accumulate the free bases. Great variation in PA composition was observed between local populations of C. pumila. The PA profiles were established for larvae and adults of E. acrea that as larvae had fed on an artificial diet supplemented with crotalaria-powder and of G. geneura fed with S. longilobus. In both experiments, the larvae had a free choice between the respective PA source and diet or food plants free of PAs. The profiles compiled for the two species reflect the alkaloid profiles of their PA sources with one exception, subulacine could never be detected in E. acrea. Besides acquired PAs, insect PAs synthesized from acquired necine bases and necic acids of insect origin were detected in the two arctiid species. These insect PAs that do not occur in the larval food sources accounted for some 40-70% (E. acrea) and 17-37% (G. geneura) of total PAs extracted from the insects. A number of novel insect PAs were identified. Plant-acquired and insect PAs were found to accumulate as N-oxides. The results are discussed in relation to specific biochemical, electrophysiological, and behavioral mechanisms involved in PA sequestration by arctiids.     

Direct Evidence for Membrane Transport of Host-Plant-Derived Pyrrolizidine Alkaloid N-Oxides in Two Leaf Beetle Genera

The chrysomelid leaf beetles Longitarsus jacobaeae, Oreina cacaliae, and O. speciosissima sequester pyrrolizidine alkaloids from their asteracean host plants and store them as nontoxic N-oxides. Previous analyses showed that Longitarsus is able to N-oxidize protoxic tertiary PAs, but did not resolve in which form N-oxides are taken up. For Oreina, beetles seem able to directly transmit the polar PA N-oxides from the gut into the hemolymph and prevent any reduction of them in the gut yielding protoxic free bases. Here, we confirm the predicted direct uptake of PAs as N-oxides by Oreina, and elucidate the situation for Longitarsus by applying double-labeled [14C]senecionine [18O]N-oxide as tracer. The beetles were fed with the tracer and subsequently senecionine N-oxide was recovered from the defensive secretions (Oreina) and beetle extracts (Longitarsus), purified by HPLC, and submitted to ESI-MS, GC-MS, and analysis of the specific radioactivity. The 18O-label is retained without any loss in the labeled senecionine N-oxide recovered from the two Oreina species. Analysis of the Longitarsus experiment was complicated by a contamination of the HPLC-purified senecionine N-oxide with a second compound, identified as a dihydrosenecionine N-oxide by high-resolution CID analysis. The dihydrosenecionine N-oxide, probably the 15,20-dihydro derivative, constitutes a major idiosyncratic senecionine metabolite present in the beetle. The recovered senecionine N-oxide retained 74% 18O-label. The remaining 25% is mostly due to loss of 18O by reduction and subsequent re-N-oxidation. The experiments confirm for both beetle genera a direct uptake of the polar nontoxic PA N-oxides, which requires specific membrane carriers. Accumulation of detrimental free base PA is prevented by glucosylation (Oreina) or N-oxidation (Longitarsus).     

Host-plant acceptance by geographic populations of the colorado potato beetle,Leptinotarsa decemlineata

We used a detailed analysis of feeding behavior to investigate the role of solanaceous alkaloids as sensory-based feeding deterrents for the Colorado potato beetle,Leptinotarsa decemlineata (Say). Experiments were conducted on three geographic, host-adapted populations of beetles to determine whether evolutionary changes in host use have been accompanied by behavioral adaptations to alkaloids. Solanine and tomatine, steroidal glycoalkaloids found in two regional host plants, did not reduce leaf consumption or significantly alter behavior patterns of newly emerged beetles, including those from populations that normally will not feed on plants containing the compounds. Atropine, a tropane alkaloid found in several taxonomically related nonhost species, caused a significant increase in sampling behavior (indicating direct action on the sensory system) and reduced acceptance of treated potato leaves. We propose that variable acceptance of host plants among regional populations ofL. decemlineata has evolved independently of adaptations to alkaloids at the sensory level. To establish that secondary compounds such as atropine influence host choice in nature, field observations are needed to confirm that beetles routinely encounter, and sample, nonhost species.     

The “Raison D'être” of pyrrolizidine alkaloids inCynoglossum officinale: Deterrent effects against generalist herbivores

In this study we tested whether pyrrolizidine alkaloids (PAs) ofCynoglossum officinale serve as antifeedants against herbivores. Total PA N-oxide extracts of the leaves significantly deterred feeding by generalist herbivores. Specialist herbivores did not discriminate between food with high and low PA levels. Three PAs fromC. officinale, heliosupine, echinatine, and 3-acetylechinatine, equally deterred feeding by the polyphagous larvae ofSpodoptera exigua. Although the plants mainly contain PAs in their N-oxide form, reduced PAs deterred feeding byS. exigua more efficiently than PA N-oxides. On rosette plants, the monophagous weevilMogulones cruciger significantly consumed more of the youngest leaves, which had the highest PA level and the highest nitrogen percentage. Larvae ofEthmia bipunctella, which are oligophagous within the Boraginaceae, did not discriminate between leaves. All generalist herbivores tested significantly avoided the youngest leaves with the highest PA levels. In the field, the oldest leaves also were relatively more damaged by herbivores than the youngest leaves. It is hypothesized that the skewed distribution of PAs over the leaves of rosette plants reflects optimal defense distribution within the plant.     

Distribution of autogenous and host-derived chemical defenses inOreina leaf beetles (Coleoptera: Chrysomelidae)

The pronotal and elytral defensive secretions of 10Oreina species were analyzed. Species feeding on Apiaceae, i.e.,O. frigida andO. viridis, or on Cardueae (Asteraceae), i.e.,O. bidentata, O. coerulea, andO. virgulata, produce species-specific complex mixtures of autogenous cardenolides.O. melanocephala, which feeds onDoronicum clusii (Senecioneae, Asteraceae), devoid of pyrrolizidine alkaloids (PAs) in its leaves, secretes, at best, traces of cardenolides. Sequestration of host-plant PAs was observed in all the other species when feeding on Senecioneae containing these alkaloids in their leaves.O. cacaliae is the only species that secretes host-derived PA N-oxides and no autogenous cardenolides. Differences were observed in the secretions of specimens collected in various localities, because of local differences in the vegetation. The other species, such asO. elongata, O. intricata, andO. speciosissima, have a mixed defensive strategy and are able both to synthesize de novo cardenolides and to sequester plant PA N-oxides. This allows a great flexibility in defense, especially inO. elongata andO. speciosissima, which feed on both PA and non-PA plants. Populations of these species were found exclusively producing cardenolides, or exclusively sequestering PA N-oxides, or still doing both, depending on the local availability of food-plants. Differences were observed between species in their ability to sequester different plant PA N-oxides and to transform them. Therefore sympatric species demonstrate differences in the composition of their host-derived secretions, also resulting from differences in host-plant preference. Finally, within-population individual differences were observed because of local plant heterogeneity in PAs. To some extent these intrapopulation variations in chemical defense are tempered by mixing diet and by the long-term storage of PA N-oxides in the insect body that are used to refill the defensive glands.     

Sequestration and metabolism of protoxic pyrrolizidine alkaloids by larvae of the leaf beetle Platyphora boucardi and their transfer via pupae into defensive secretions of adults

Several neotropical leaf-beetles of the genus Platyphora ingest and specifically metabolize plant acquired pyrrolizidine alkaloids (PAs) of the lycopsamine type (e.g., rinderine or intermedine) and enrich the processed alkaloids in their exocrine defensive secretions. In contrast to the related palaearctic leaf beetles of the genus Oreina, which absorb and store only the non-toxic alkaloid N-oxides, Platyphora sequesters PAs exclusively as protoxic tertiary amines. In this study, the ability of P. boucardi larvae to accumulate PAs was investigated. Tracer studies with [¹⁴C]rinderine and its N-oxide revealed that P. boucardi larvae, like adult beetles, utilize the two alkaloidal forms with the same efficiency, but accumulate the alkaloid as a tertiary amine exclusively. Ingested rinderine is rapidly epimerized to intermedine, which is localized in the hemolymph and all other tissues; it is also detected on the larval surface. Like adults, larvae are able to synthesize their own alkaloid esters (beetle PAs) from orally administered [¹⁴C]retronecine and endogenous aliphatic 2-hydroxy acids. These retronecine esters show the same tissue distribution as intermedine. A long-term feeding experiment lasting for almost four months revealed that retronecine esters synthesized from [¹⁴C]retronecine in the larvae are transferred from larvae via pupae into the exocrine glands of adult beetles. Pupae contain ca. 45% of the labeled retronecine originally ingested, metabolized, and stored by larvae; ca. 12% of larval radioactivity could be recovered from the defensive secretions of adults sampled successively over two and a half months. Almost all of this radioactivity is found in the insect-made retronecine esters that are highly enriched in the defensive secretions, i.e., more than 200-fold higher concentration compared to pupae.     

Production of cardenolides versus sequestration of pyrrolizidine alkaloids in larvae ofOreina species (Coleoptera,Chrysomelidae)

Adult leaf beetles of the genusOreina are known to be defended either by autogenously produced cardenolides or by pyrrolizidine alkaloids (PAs) sequestered from the food plant, or both. In this paper we analyze larvae of differentOreina species and show that the larvae contain the same defensive toxins as the adults in quantities similar to those released in the adults' secretion. Both classes of toxins are found in the body and hemolymph of the larvae, despite their different origins and later distribution in the adults. Larvae of sequestering species differed in their PA patterns, even though they fed on the same food plants. The concentration in first-instar larvae of a PA-sequestering species was similar to that in fourth-instar larvae. In all stages examined, the amount of PAs per larva did not greatly exceed the estimated uptake of one day. Eggs of two oviparous species contained large concentrations of the adult's toxins, while neonates of a sequestering larviparous species had no PAs.     

Trirhabda canadensis (Coleoptera: Chrysomelidae) responses to plant odors

The responses of the goldenrod leaf beetleTrirhabda canadensis to host and nonhost volatile odors were tested in a Y-tube olfactometer in the laboratory. Beetles preferred host to nonhost odors and were sensitive to concentrations of host odor. Beetles distinguished between host and nonhost volatiles of only one of the two nonhostSolidago species; host volatiles were preferred to all nonhost volatiles at the family and order levels. In other words, all nonhosts above the genus level had similar effects on beetle responses. Although the odors of most nonhosts were neutral (i.e., neither attractive nor repellent) to the beetles as tested against air, this neutrality disappeared if the odors of two or more nonhosts were added to the host odor and beetles were given a choice between this mixture and pure host odor. Given this choice, they strongly preferred pure host odor, which suggests that diversity of odors per se is unattractive to the beetles. Beetles walked rather than flew to locate their hosts in the field, and their movements suggest that they used olfactory cues to locate hosts.     

Pyrrolizidine alkaloids in the arctiid mothHyalurga syma

The arctiid mothHyalurga syma (subfamily Pericopinae) sequesters pyrrolizidine alkaloids (PAs) from its larval food plantHeliotropium transalpinum (Boraginaceae). Colorimetric quantification of total PAs in the larvae, pupae, and adults ofHyalurga revealed mean values of about 286–445g per individual (1.4–2.6% of dry weight). The PA mixtures found in the moth and its larval food plant were evaluated by GC-MS. Food-piant leaves were found to contain the diastereoisomeric retronecine esters indicine (IIIa), intermedine (IIIb), and lycopsamine (IIIc), and the heliotridine ester rinderine (IIId) only as minor constituents, whereas 3-acetylrinderine (IVc) (68% of total PAs) and the respective 3-acetyl esters of indicine (IVa) and intermedine (IVb) (both 17%) were the major alkaloids. Supinine (IIa) is detectable in traces only. The PA mixtures in eggs, larvae, pupae, and imagines ofHyalurga were identical: indicine, intermedine, and lycopsamine accompanied by considerable amounts of supinine and amabiline or coromandalinine (IIb/IIc) were the major components. Only larvae were found to store small quantities of a 3-acetyl derivative. Rinderine and its 3-acetyl ester were never found in the insects. Low concentrations of the arctiidspecific PA callimorphine (I) were present in larvae, pupae, and imagines. The differences in the PA patterns of the insects and their larval food plant suggest thatHyalurga is capable of modifying plant-derived PAs by inversion of the 7-OH configuration (conversion of the necine base heliotridine into retronecine), and perhaps the inversion of the 3-OH [conversion of (+)-trachelanthic acid into (–)-viridifloric acid], although the possibility of a selective sequestration of the respective retronecine esters cannot be excluded. Some trials with the orb-weaving spiderNephila clavipes, a common neotropical predator, showed that both freshly emerged and field-caught adults ofHyalurga syma are liberated unharmed by the spider. The liberation could be related to the presence of PAs in the moths.     

Volatile compounds induced by herbivory act as aggregation kairomones for the Japanese beetle (Popillia japonica Newman)

The Japanese beetle is a polyphagous insect that typically aggregates on preferred host plants in the field. We studied the response of Japanese beetles to artificial damage, fresh feeding damage, and overnight feeding damage to test the hypothesis that beetles are attracted to feeding-induced volatiles. Crabapple leaves that had been damaged overnight by Japanese beetles or fall webworms attracted significantly more Japanese beetles than did undamaged leaves. Artificially damaged leaves or leaves freshly damaged by Japanese beetles, however, were not significantly more attractive than undamaged leaves. Leaves that had been damaged overnight by Japanese beetles or fall webworms produced a complex mixture of aliphatic compounds, phenylpropanoid-derived compounds, and terpenoids. In comparison, artificially damaged leaves or leaves with fresh Japanese beetle feeding damage generated a less complex blend of volatiles, mainly consisting of green-leaf odors. Feeding-induced odors may facilitate host location and/or mate finding by the Japanese beetle.     

Phloeophagous and predaceous insects responding to synthetic pheromones of bark beetles inhabiting white spruce stands in the Great Lakes region

Tree killing and saprophytic bark beetles exert important ecological and economic roles in North American spruce forests. Chemical signaling among bark beetles, and responses by associate insects such as predators and competitors, have significant effects on the population dynamics and ecology of this community. Synthetic pheromones of primary (tree killing) and secondary (saprophytic) bark beetle species and blank controls were tested using multiple funnel and lower stem flight traps in white spruce forests in Wisconsin, Michigan, and Minnesota. Six phloeophagous and four predaceous species were collected with significant attraction by the bark beetles Dryocoetes affaber, Dryocoetes autographus, and Polygraphus rufipennis, and the predatory checkered beetles (Coleoptera: Cleridae) Thanasimus dubius and Enoclerus nigrifrons. In general, trap catches to synthetic lures resembled the species composition obtained by felling trees and collecting emerging beetles in a companion study, although several species showed differing trends. Some cross attraction occurred among bark beetles and between bark beetles and predatory beetles. For example, P. rufipennis was abundant in traps baited with Dryocoetes spp. pheromones. Thanasimus dubius and E. nigrifrons were collected in significant numbers in traps baited with the pheromone of the spruce beetle (Dendroctonus rufipennis), frontalin plus -pinene. This is a new observation for E. nigrifrons. Attraction of T. dubius to the pheromones of at least three bark beetle species in the Great Lakes region, as well as to several southern and western species, reflects its role as a habitat specialist and feeding generalist. Several other important predators and competitors commonly obtained in pine forests in this region were not obtained in these spruce stands, either in response to synthetic pheromones of spruce colonizing beetles, or in host material colonized by these beetles. Potential differences in predator prey dynamics between spruce and pine ecosystems in the Great Lakes region are discussed.     

Responses of Bark Beetle-Associated Bacteria to Host Monoterpenes and Their Relationship to Insect Life Histories

Bark beetles that colonize living conifers and their microbial associates encounter constitutive and induced chemical defenses of their host. Monoterpene hydrocarbons comprise a major component of these allelochemicals, and many are antibiotic to insects, fungi, and bacteria. Some bark beetle species exhaust these defenses by killing their host through mass attacks mediated by aggregation pheromones. Others lack adult aggregation pheromones and do not engage in pheromone-mediated mass attacks, but rather have the ability to complete development within live hosts. In the former species, the larvae develop in tissue largely depleted of host terpenes, whereas in the latter exposure to these compounds persists throughout development. A substantial literature exists on how monoterpenes affect bark beetles and their associated fungi, but little is known of how they affect bacteria, which in turn can influence beetle performance in various manners. We tested several bacteria from two bark beetle species for their ability to grow in the presence of a diversity of host monoterpenes. Bacteria were isolated from the mountain pine beetle, Dendroctonus ponderosae Hopkins, which typically kills trees during colonization, and the red turpentine beetle, Dendroctonus valens LeConte, which often lives in their host without causing mortality. Bacteria from D. ponderosae were gram-positive Actinobacteria and Bacilli; one yeast also was tested. Bacteria from D. valens were Actinobacteria, Bacilli, and γ-Proteobacteria. Bacteria from D. valens were more tolerant of monoterpenes than were those from D. ponderosae. Bacteria from D. ponderosae did not grow in the presence of α-pinene and 3-carene, and grew in, but were inhibited by, β-pinene and β-phellandrene. Limonene and myrcene had little inhibitory effect on bacteria from either beetle species. Tolerance to these antibiotic compounds appears to have resulted from adaptation to living in a terpene-rich environment.     

Effects of Host Tree Species on Attractiveness of Tunneling Pine Engravers, Ips pini, to Conspecifics and Insect Predators

The effect of host tree species on the attractiveness of tunneling Ips pini to flying beetles and their insect predators in Wisconsin was investigated. Tree species influenced the flight response of both predators and prey in the same rank order. Ips pini and its major predators, Thanasimus dubius and Platysoma cylindrica, were more attracted to I. pini males boring into bark–phloem disks of Pinus strobus L. than Pinus banksiana Lamb, and least attracted to I. pini males boring into bark–phloem disks of Pinus resinosa. Sources of within-tree, between-tree, and between-species variation in the degree of attraction elicited by tunneling beetles were quantified. A bioassay for evaluating host tree effects on pheromone based communication among bark beetles under conditions of controlled beetle entry was developed. Possible mechanisms of host species effects on the dynamics of predator and prey interactions in bark beetle ecology are discussed.     

Insect-synthesised Retronecine Ester Alkaloids: Precursors of the Common Arctiine (Lepidoptera) Pheromone Hydroxydanaidal

Many pyrrolizidine alkaloid (PA)-adapted insects convert PAs sequestered from their larval host plants into “insect-PAs” in which the acid components of the alkaloids are replaced by small, branched aliphatic 2-hydroxy acids of insect origin. It has been proposed that insect-PAs are precursors of the pheromone hydroxydanaidal in male Estigmene acrea moths, but it is not clear why they are specifically required or what the structural features or chemical properties are that make insect-PAs more suitable for conversion into hydroxydanaidal than superficially similar alkaloids of plant origin. Evidence is presented that insect-PAs are also precursors of hydroxydanaidal in the polyphageous arctiine, Creatonotos transiens, and a new biosynthetic pathway to hydroxydanaidal is proposed that has a mandatory requirement for insect-PAs as intermediates.     

Pyrrolizidine alkaloids as oviposition stimulants for the cinnabar moth,Tyria jacobaeae

In choice experiments with artificial leaves, we tested related pyrrolizidine alkaloids (PAs) for their stimulatory effects on the oviposition of the cinnabar moth, a specialist on the PA-containing plant Senecio jacobaea. The PAs from S. jacobaea that we tested stimulated oviposition. Monocrotaline also stimulated oviposition although this PA is not found in plants of the genus Senecio. The moths preferred ovipositing on filter paper with a PA mixture extracted from S. jacobaea to ovipositing on filter paper with single PAs. Senkirkine, heliotrine, and retrorsine did not stimulate oviposition. The nonactive retrorsine differs only in one OH group to the active senecionine, indicating that small structural differences alter the stimulatory activity of PAs. However, a PA mixture extracted from a nonhost plant, Senecio inaequidens, that consisted of 81% of the nonactive retrorsine did stimulate oviposition. Oviposition preferences between Senecio species seem to be determined by chemical compounds other than PAs.     

Colorado Potato Beetle Toxins Revisited: Evidence the Beetle Does Not Sequester Host Plant Glycoalkaloids

The Colorado potato beetle feeds only on glycoalkaloid-laden solanaceous plants, appears to be toxic to predators, and has aposematic coloration, suggesting the beetle may sequester alkaloids from its host plants. This study tested 4th instars and adults, as well as isolated hemolymph and excrement, to determine if the beetles sequester, metabolize, or excrete alkaloids ingested from their host plants. HPLC analysis showed: that neither the larvae nor the adults sequestered either solanine or chaconine from potato foliage; that any alkaloids in the beetles were at concentrations well below 1 ppm; and that alkaloids were found in the excrement of larvae at approximately the same concentrations as in foliage. Analysis of alkaloids in the remains of fed-upon leaflet halves plus excreta during 24 hr feeding by 4th instars, as compared to alkaloids in the uneaten halves of the leaflets, showed that equal amounts of alkaloids were excreted as were ingested. The aposematic coloration probably warns of a previously-identified toxic dipeptide instead of a plant-derived alkaloid, as the Colorado potato beetle appears to excrete, rather than sequester or metabolize, the alkaloids from its host plants.     

Variation in cardenolide content of the lygaeid bugs,Oncopeltus fasciatus andLygaeus kalmii kalmii and of their milkweed hosts (Asclepias spp.) in central California

A colorimetric assay was used to quantify the amount of cardenolides in the lygaeid bugsOncopeltus fasciatus andLygaeus kalmii kalmii and their milkweed host plants (Asclepias spp.) in central California. The cardenolide content of individual insects, determined in microgram equivalents of digitoxin, varied from zero to over 300 g per insect. Sources of variation of cardenolide content in the insects include interspecific and intraspecific differences in the content of the host plant species and also differences in the content of plant organs on which insects were feeding. This last source of variability may explain temporal variation in the cardenolide content of the insects. Adults ofO. fasciatus, which migrate into California in the late spring and early summer, and adults ofL. k. kalmii, which emerge from winter hibernacula in the early spring, contained small to immeasureable amounts of cardenolides. The colonization pattern ofO. fasciatus on species ofAsclepias in north central California suggests that this species does not maximize its opportunities to sequester large quantities of cardenolides from potential hosts. The emetic potential of lygaeids in California to vertebrate predators is briefly discussed.     

Taste receptors for pyrrolizidine alkaloids in a monophagous caterpillar

Caterpillars of Utetheisa ornatrix are monophagous on species of Crotalaria from which they obtain pyrrolizidine alkaloids (PAs) for defense and which the males convert to a pheromone. We show that a taste receptor neuron in each of the lateral and medial galeal styloconic sensilla responds to PAs of three different types. Monocrotaline, commonly present in Crotalaria species, is the most strongly stimulating with thresholds of response below 10^–11 and 10^–9 M in the two sensilla. These are among the lowest known taste thresholds in any insect and are similar to the thresholds for PAs in a polyphagous arctiid caterpillar that also sequesters PAs and uses them as the source of male pheromone. The receptors also respond to heliotrine, a type of PA that is probably never encountered by the insects. Monocrotaline and senecionine N-oxide are shown in behavioral assays to be phagostimulants. The data show that there is no tight link between taste receptor sensitivity to specific PAs and hostplant selection in these caterpillars. Perhaps the adults are primarily responsible for hostplant selection.     

Disruption of Secondary Attraction of the Spruce Beetle, Dendroctonus rufipennis, by Pheromones of Two Sympatric Species

Capture of spruce beetles, Dendroctonus rufipennis, in multiple-funnel traps baited with frontalin and -pinene was reduced by up to 42% in the presence of synthetic (+)-exo- and (+)-endo-brevicomin, aggregation pheromones of the sympatric species Dryocoeles affaber. (±)-endo-Brevicomin was inhibitory to spruce beetles in two experiments and (±)-exo-brevicomin was inhibitory in one experiment, reducing spruce beetle trap catches by up to 87% and 75%, respectively. Spruce beetle trap catches were also reduced by 85% by (±)- or (+)-ipsdienol, but not by (–)-ipsdienol. Ips tridens, a second sympatric species, produces both enantiomers of ipsdienol in its pheromone blend. Responses by D. affaber to its own pheromone were significantly enhanced by addition of the spruce beetle lure. Enantiospecific pheromones of secondary competing species, or less costly racemic substitutes, may be useful for managing spruce beetles using competitive displacement or exclusion. Baiting susceptible hosts with pheromones of secondary species may enhance attack by secondary species, while partially repelling spruce beetles.     

A potential kairomone stimulates pheromone-responsive receptor neurons inUtetheisa ornatrix (Lepidoptera: Arctiidae)

The olfactory receptor neurons in basiconic sensilla on the antennae ofUtetheisa ornatrix, which, in females, had earlier been shown to be responsive to stimulation with hydroxydanaidal (HD), are here shown to be responsive to volatile substances in samples of pyrrolizidine alkaloids (PAs). These latter substances are secondary plant metabolites present in the host plant of the larvae. Their sequestration during larval life serves to protect all life stages from predation. In males, the PAs also provide precursors for the production of the male pheromone, HD. In females, basiconic receptor neurons begin to respond to stimulus cartridges containing 1 ng of (R)-(–)-hydroxydanaidal, 100 ng of its isomer, (S)-(+)-hydroxydanaidal, and to volatiles emanating from 10g of the alkaloids monocrotaline and heliotrine. Receptor neurons in males are generally responsive to the same array of substances but with reduced sensitivity. The dietary background of the adult moths with respect to prior ingestion of PAs does not influence the response capabilities of basiconic receptor neurons to HD or to the volatile components of PAs. Earlier studies in another arctiid (Rhodogastria), had indicated that trace amounts of HD are present in PAs, presumably as their hydrolysis product. Thus we assume that, under natural conditions, HD may be an active component of the volatiles from PAs and may serve as both a male pheromone and a kairomone. Sensitivity to HD could thereby provide information about the location and PA content of potential mates and food plants. Mechanisms that may have resulted in the evolution of a signaling system with these properties are discussed.