Behavioral and electrophysiological responses of Aphids to host and nonhost plant volatiles

Alate and apterous virginoparae ofAphis fabae Scop, and alate virginoparae ofBrevicoryne brassicae (L.), walking in a linear track olfactometer, were attracted by odor from leaves of their host plants.A. fabae responded to odor from undamaged but not damaged bean leaves. Gynoparae (autumn migrants) ofA. fabae, however, did not respond to their host plant (spindle,Euonymus europaeus) odor. Odors of certain nonhost plants masked the attractiveness of the host plant leaves, but tansy (Tanacetum vulgare) and summer savory (Satureja hortensis) volatiles repelledB. brassicae andA. fabae, respectively. 3-Butenyl isothiocyanate attractedB. brassicae andLipaphis erysimi (Kalt.), the latter species being more sensitive in both behavioral and electrophysiological studies. Isothiocyanate receptors were found on the antennae ofA. fabae, which was repelled by these compounds, 4-pentenyl isothiocyanate being the most active.     

Behavioral and electrophysiological responses of the banana weevilCosmopolites sordidus to host plant volatiles

Male and femaleCosmopolites sordidus were attracted to freshly cut banana rhizome and pseudostem in a still-air olfactometer. Females responded similarly to odors from a comparatively resistant and from a susceptible cultivar of banana, when presented as either freshly cut tissue or as Porapak-trapped volatiles. Females were also attracted to rotting banana pseudostem and to volatiles collected from it. Males and females gave similar responses to host tissue in both the behavioral bioassay and to collected volatiles in EAG recordings. Weevils did not respond, either behaviorally or electrophysiologically, to a synthetic mixture of mono- and sesqiterpenes, which made up over 9% of the volatiles collected from pseudostem.     

Oviposition stimulants for the beetle,Monochamus alternatus hope,in inner bark of pine

Field and laboratory ovipositional responses ofMonochamus alternatus Hope, respectively, to methanol and water extracts from pine inner bark were examined in comparison with those to pine inner bark, especially using a laboratory-built apparatus for the latter bioassay. Irrespective of the existence of volatiles from paraquat-induced lightwood, pine inner bark and its methanol and water extracts stimulated ovipositional response only in the presence of free moisture.     

Weak responses of pine sawfly larvae to high needle flavonoid concentrations in scots pine

Responses of sawfly larvae (Hymenoptera, Diprionidae) to the flavonoid taxifolin glucoside in their host plant were studied in a laboratory experiment. Larvae ofNeodiprion sertifer andDiprion pini were raised from egg hatch to cocoon spinning on two Scots pine (Pinus sylvestris) chemotypes, one without needle taxifolin glucoside (–) and the other containing 2–4% taxifolin glucoside (+). The (+) chemotype had somewhat lower concentrations of needle terpenoids (resin acids) than the (–) chemotype. Current-year needles had higher taxifolin glucoside concentrations than mature needles. There were no differences in survival or body size betweenN. sertifer larvae that fed on the (+) chemotype and those that fed on the (–) chemotype. FemaleD. pini larvae raised on (+) needles developed 6% more slowly than larvae fed (–) needles. The results from this study are contrary to earlier findings showing that flavonoid glucosides have strong negative effects on insect performance. Possible explanations for the different outcomes are discussed.     

Behavioral responses to own and other species' scent marks inLemur fulvus andLemur macaco

Behavioral responses (duration and frequencies of sniffing, scent marking activity) were studied in one captive group each ofL. fulvus andL. macaco when confronted with their own and other species' scent marks. Both species showed less interest in neutral objects or objects carrying their own group's odor than odors from an unfamiliar group of the same species or from another lemur species. The persistence of scent marks was also investigated.L. fulvus scent appeared to possess greater longevity thanL. macaco scent. Olfactory long-term memory was analyzed inL. fulvus. The results showed that familiar rejected group members could be discriminated from an unfamiliar conspecific through odor cues 10 months after their eviction from the group.     

Behavioral and electrophysiological responses of natural enemies to synomones from tea shoots and kairomones from tea aphids,Toxoptera aurantii

Olfactometer bioassays and electrophysiological studies showed that the lacewing, Chrysopa sinica, the aphid parasitoid, Aphidius sp., and the coccinellid, Coccinella septempunctata, all responded to volatiles from tea aphids, Toxoptera aurantii, to hexane or ether rinses of tea aphid cuticles, and to synomones released by aphid-damaged tea shoots, as well as to the tea shoot–aphid complex. Each natural enemy spent more time searching on a filter paper treated with tea aphid honeydew than on a blank control filter paper. The interaction between synomones from aphid-damaged shoots and kairomones from tea aphids enhanced the responses to the plant–host complex. There was a significant, logistic dose–response relationship between the number of natural enemies responding and the odor stimulus concentration. Volatile components from the plant–host complex, obtained by air entrainment, were identified by their mass spectra and retention times and confirmed by comparison with standard samples. These were (Z)-3-hexen-1-ol, benzaldehyde, (E)-2-hexenal, (Z)-3-hexenyl acetate, ocimene, linalool, geraniol, indole, and (E)-2-hexenoic acid. The main components in a hexane rinse from tea aphid cuticle were benzaldehyde, undecane, 2,5-hexanedione, 2,5-dihydrothiophene, linalool, 4-methyl-octane, and eicosane, whereas the main components from an ether rinse were (E)-2-hexenoic acid, heptadecane, pentadecane, eicosane, tetratetracontane, and nonadecane. Benzaldehyde elicited the strongest responses from natural enemies in the olfactometer and the largest electroantennogram (EAG) responses. While the amount of odor was small, Coccinella septempunctata was slightly more sensitive than Chrysopa sinica and Aphidius sp. An increase in doses of benzaldehyde, (E)-2-hexenal, and (Z)-3-hexenyl acetate caused the EAG responses of each natural enemy to decrease. When the doses of (Z)-3-hexen-1-ol, linalool, and geranoil increased, EAGs of Chrysopa sinica and Aphidius sp. increased, but EAGs of Coccinella septempunctata decreased. When the dose of indole increased, EAGs of Coccinella septempunctata decreased, but those of Aphidius sp. increased. This study demonstrates that tea shoot–aphid complexes emit volatile synomones, while the odors from tea aphids, aphid cuticle extracts, and tea aphid honeydew contain kairomones, to which the natural enemies show a logistic dose–response.     

Mite predator responses to prey and predator-emitted stimuli

We found that the searching behavior of two acarine predators,Amblyseius fallacis andPhytoseiulus macropilis, for prey,Tetranychus urticae, is affected by the following stimuli: (1) prey silk and associated feces, whose combined physical and chemical properties elicit reduction in the rate of predator movements and longer halts; (2) kairomone extracted from prey silk and associated feces, which, upon contact, elicits frequent predator return to prey-inhabited locales; and (3) predator-emitted marking pheromone, which elicits shorter duration of search in presearched prey locales. We also found that treatment of filter paper with prey kairomone or silk enhanced predator location of prey eggs, leading us to speculate that application of synthetic prey kairomone could be useful in pest management programs.     

Olfactory responses ofIps plastographus maritimus lanier (coleoptera: Scolytidae) to insect and host-associated volatiles in the laboratory

Attraction of both sexes ofIps plastographus maritimus Lanier to bark-phloem-xylem discs of Monterey pine,Pinus radiata D. Don, was demonstrated in the laboratory. Increasing concentrations of male and female volatiles trapped separately and released in a one-to-one ratio decreased attraction for both sexes combined. Attraction of both sexes to volatiles derived from males and females tunneling together in a one-to-one ratio increased with increasing concentration of extract. Attraction of males and females to male-infested discs and to trapped male volatiles increased with increasing dose of males or male extract. Attraction of males and females to female-infested discs and to trapped female volatiles was also demonstrated. The presence of females in male galleries reduced the attractiveness of infested disks to both sexes combined. Increasing numbers of females, tunneling separately from males in the same disc, reduced attraction of males, but not females. When a constant attractive dose of male volatiles was released with increasing doses of female volatiles, there was no difference in response of either sex when female volatiles were present compared with the response to male volatiles alone. When a constant attractive dose of male volatiles was released with increasing concentrations of volatiles derived from males and females tunneling together in a one-to-one ratio, attraction ofI. p. maritimus decreased. Response of females was frequently higher than that of males to the same attractant source. Hence, both sexes produce an attractant, and both sexes tunneling together in the same gallery reduce attraction of males and females to an attractive dose of male attractant.     

Behavioral responses of elm bark beetles to baited and unbaited elms killed by cacodylic acid

Diseased elms, treated with various doses of cacodylic acid in northwest England, became attractive to elm bark beetles (Coleoptera: Scolytidae). This attraction seemed to be independent of pheromone baits. However attractive the trees became, they were unsuitable to the beetles as breeding sites since significantly more beetles visited the trees than were stimulated to penetrate and attempt to breed. It seems as if colonization of trap trees by the bark saprophytePhomopsis oblonga following cacodylic acid treatment made the trees unsuitable to beetles for breeding.     

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.     

Attraction of pinyon pine bark beetle,Ips hoppingi,to conspecific andI. confusus pheromones (Coleoptera: Scolytidae)

Females of a pinyon pine bark beetle,Ips hoppingi Lanier, were less attracted by the aggregation pheromone produced by conspecific males than by the pheromone produced by the neighboring sibling species,I. confusus (LeConte). Cross-attraction was elicited by males infesting the regional pinyon pine hosts (P. discolor andP. edulis) of eitherIps species in south-eastern Arizona. Pheromonal specificity has not accompanied speciation in this species pair.     

Antennal responses of the two host races of the larch bud moth,Zeiraphera diniana,to larch and cembran pine volatiles

The larch bud moth (LBM) Zeiraphera diniana Guenée causes defoliation on larch in the Alps at 8- to 10-year intervals, after which populations crash. There are two LBM host races, one on larch and the other on cembran pine. These host races are morphologically indistinguishable as adults but they differ genetically in larval color types. Furthermore, females of each host race produce distinct pheromone blends and show oviposition preferences for their respective hosts. It is not clear to what extent host choice contributes to assortative mating in the LBM. Here, we compare the olfactory sensitivities of the two host races to the odors of fresh foliage of the host plants using the electroantennogram (EAG) technique, and the responses of the two host races to volatiles collected from the two host plants as analyzed by gas-chromatography-linked antennographic detection (GC-EAD). Both sexes of the larch and cembran host races show the same EAG responses to vapors of fresh larch and cembran pine foliage. Fifteen plant volatiles identified as chemostimuli by GC-EAD from larch and cembran pine odors elicited the same antennogram responses from the two host races. However, the GC-EAD analyses indicate that the number and quantity of chemostimuli emanating from each host plant is different. It is, therefore, most probably the array of olfactory receptors responding to the bouquet of volatiles unique to each host plant that underlies the host preferences of the two races. What remains open is the extent to which the similarity of the olfactory systems may contribute to cross-attraction. The fact that LBM individuals with intermediate characteristics between the two host races exist, suggests that olfactory perception does not hinder gene flow and contributes to sustained genetic diversity within the species Z. diniana.     

Behavioral and electrophysiological study of antifeedant mechanisms associated with polyhydroxy alkaloids

Eleven polyhydroxy alkaloids of plant origin were tested for anti-feedant effects against larvae of the lepidopteransSpodoptera littoralis, Spodoptera frugiperda, Heliothis virescens, andHelicoverpa armigera. Data from behavioral and electrophysiological investigations were correlated to reveal information on the mode of action of the antifeedants. The pyrrolidine DMDP was an effective antifeedant for all four species, whereas the piperidines fagomine and XZ-1 and the pyrrolizidine alexine were all ineffective as antifeedants. The activity of the pyrrolidines CYB-3 and DAB-1, the piperidines DNJ, DMJ, and BR1, and the bicyclic octahydroindolizine castanospermine varied among species. The investigation focuses on the structural similarities between some of the alkaloids and some common phagostimulatory sugars and illustrates a neural interaction involving the neurons that are differentially responsive to alkaloids and sugars. InS. littoralis, the neurons responding specifically to the alkaloids DMDP, DAB-1, and castanospermine and to the sugars fructose, sucrose, and glucose are more active when the compounds are applied singly than when an alkaloid and a sugar are applied together. The implications for the occurrence and functioning of different sugar receptor sites are discussed.     

Behavioral responses of the parsnip webworm to host plant volatiles

Sixth instars of the parsnip webworm, Depressaria pastinacella, orient by olfaction to bud, male flowers, and female flowers of their primary host plant, Pastinaca sativa. Because octyl acetate and octyl butyrate are characteristic of tissues consumed by the sixth instar, we investigated the influence of these esters on webworm feeding behavior and chemo-orientation. Although octyl acetate and octyl butyrate are feeding deterrents, and octyl butyrate is an olfactory repellent, octyl acetate serves as an olfactory attractant. In olfactometers, webworms do not show a preference when given a choice between octyl acetate and host plant tissues. These findings suggest that octyl acetate is a sufficient cue for olfactory orientation. Such behavior may explain differences in the relative abundance of these esters observed among populations of wild parsnip under differential selection pressure from these insects.     

Feeding response of Ips paraconfusus to phloem and phloem metabolites of Heterobasidion annosum-inoculated ponderosa pine,Pinus ponderosa

In studies of feeding by the bark beetle, Ips paraconfusus, two pine stilbenes (pinosylvin and pinosylvin methyl ether), ferulic acid glucoside, and enantiomers of the four most common sugars present in ponderosa pine phloem (sucrose, glucose, fructose, and raffinose) did not stimulate or reduce male feeding when assayed on wet -cellulose with or without stimulatory phloem extractives present. When allowed to feed on wet -cellulose containing sequential extracts (hexane, methanol, and water) of ponderosa pine phloem, methanol and water extractives stimulated feeding, but hexane extractives did not. Males confined in wet -cellulose containing aqueous or organic extracts of culture broths derived from phloem tissue and containing the root pathogen, Heterobasidion annosum, ingested less substrate than beetles confined to control preparations. In an assay using logs from uninoculated ponderosa pines, the mean lengths of phloem in the digestive tracts increased as time spent feeding increased. Males confined to the phloem of basal logs cut from ponderosa pines artificially inoculated with H. annosum ingested significantly less phloem than beetles in logs cut from trees that were (combined) mock-inoculated or uninoculated and did not contain the pathogen. However, individual pathogen-containing treatments were not significantly different from uninoculated controls. It was concluded that altered feeding rates are not a major factor which may explain why diseased ponderosa pines are colonized by I. paraconfusus.     

Effects of Enantiomeric Blend of Verbenone on Response ofIps paraconfusus to Naturally Produced Aggregation Pheromone in the Laboratory

The aggregation pheromone produced by maleIps paraconfusus (Coleoptera: Scolytidae) tunneling in Monterey pine,Pinus radiata, logs was trapped on Porapak Q. A concentration of an extract of trapped volatiles that attracted beetles was determined in the laboratory through a concentration–response analysis of walking behavior of males and females. The interruptant effects of four concentrations of verbenone on response to a constant concentration of this naturally produced aggregation pheromone were tested with males and females. Independent of its enantiomeric composition [99.5% (S)-(–), 93.1 % (S)-(–), and 98.3% (R)-(+)], verbenone significantly reduced the percentage of females (but not males) reaching the attractant source. However, when the time required for beetles of both sexes taken together to reach the attractant source was considered, verbenone of higher enantiomeric purity had a greater effect on beetle behavior. Solutions of 99.5% (S)-(–)- and 98.3% (R)-(+)-verbenone increased the time required for beetles to reach the attractive source when compared to 93.1% (S)-(–)-verbenone. When pooled across enantiomeric blends, increasing concentrations of verbenone resulted in slower responses in beetles that reached the attractant source within 2 min. Males and females did not respond to verbenone alone.     

Antennnal olfactory and behavioral response of southern pine beetle,Dendroctonus frontalis,to analogs of its aggregation pheromone frontalin

Antennal olfactory (electroantennogram) and laboratory and field behavioral tests were carried out on the response ofDendroctonus frontalis to its aggregation pheromone frontalin and analogs. The analogs were compounds modified by altering the position and methyl groups and/or by their deletion. Any modification to the frontalin structure significantly reduced both the antennal olfactory and behavioral response byD. frontalis. Beetle response, although significantly reduced, was elicited at the receptor level and in a laboratory bioassay by all analogs. However, only one analog (endo-5,7-dimethyl-Frontalin) elicited significant response from field populations of the beetle.Texas Agricultural Experiment Station paper TA-22507. The work was funded in part by McIntire-Stennis project 1525 and by an operating grant from the Alberta Forest Service through the Forest Development Research Trust Fund. The findings, opinions, and recommendations expressed are those of the authors and not necessarily those of the U.S. Department of Agriculture. All programs and information of the Texas Agricultural Experiment Station are available without regard to race, ethnic origin, religion, sex, or age.     

Behavioral and electrophysiological responses of the brownbanded cockroach,Supella longipalpa,to stereoisomers of its sex pheromone,supellapyrone

Females of the brownbanded cockroach, Supella longipalpa, release a sex pheromone (supellapyrone) during a calling behavior and attract males from a distance. Supellapyrone has four possible configurations resulting from two asymmetric carbons at positions 2 and 4 (i.e., 2R,4R; 2R,4S; 2S,4R; and 2S,4S), but only the RR isomer is produced by females. Using pure synthetic stereoisomers in field tests, we showed that males are attracted to RR but also to high concentrations of the isomer SR. To study the activity of the stereoisomers in more detail we developed behavioral and electroantennogram (EAG) dose–response curves for each. Behaviorally, RR was the most active isomer with just 0.3 pg delivered on a filter paper being sufficient to elicit 50% male response in the olfactometer. Males were also attracted to SR and SS in the olfactometer, but at much higher dosages (100×) than the natural compound; RS did not elicit behavioral responses at any of the doses tested. In EAG assays, the antenna of male S. longipalpa showed high and similar sensitivity to RR and SR, but a much lower (10%) sensitivity to SS and practically no response to RS. The lack of agreement between behavioral and electrophysiological data suggested either that RR and SR stimulate different antennal sensory neuron types, or that some aspect of the interaction between the pheromone and the sensillum environment or the receptor neuron itself is different. To test the first hypothesis we examined the response of the antenna before and after adaptation with each of the four stereoisomers. Positive cross-adaptation between RR and SR suggests that these two compounds stimulate the same receptor cells. Therefore, the lack of agreement between behavioral and EAG dose–response curves could be explained by isomer-specific molecular interactions between the pheromone and the receptor neuron. Although RR and SR produced the same EAG amplitude, stimulation with SR resulted in a slower recovery rate (i.e., wider peaks) than stimulation with RR. To gain further understanding of the response specificity of the antennae to the different stereoisomers we compared EAG responses (amplitude and recovery time) in response to individual stereoisomers and binary mixtures of isomers. These tests showed additive responses of the EAG amplitude to mixtures of compounds, but nonadditive responses of EAG recovery time. Therefore, peak height and width are independent parameters of the EAG, probably representing different intrasensillar events, and likely resulting in the expression of different behavioral responses.     

Biochemical responses of Pinus pinaster trees to fire-induced trunk girdling and crown scorch: secondary metabolites and pigments as needle chemical indicators

The biochemical responses to varying levels of localized heat damage to stems and crowns were studied in Pinus pinaster trees in Galicia, Spain. The objective was to ascertain the utility of secondary metabolites (total polyphenols, condensed tannins, astringency, free and esterified phenolic acids) and pigments (chlorophylls and carotenoids) as chemical indicators of localized fire damage. The study involved three levels of girdling by trunk heating (0%, 60%, and 75%), three levels of crown scorching (0%, 50%, and 75%), and all combinations of those trunk and crown treatment levels. Secondary metabolites and pigments were analyzed in needles before, during, and up to 8 months after fire treatments. High levels of polyphenols, condensed tannin, and pigments occurred in situations where the sole treatment was 75% crown scorching. Low levels of hydroxycinnamic acids were the result when the treatment was 75% trunk girdling. These responses occurred in the first two months after fire. Thereafter, normal values were observed. Low values of chlorophyll a/b ratio and high levels of free protocatechuic and esterified syringic acids were found to be long-term indicators of trunk girdling damage combined with crown scorching.     

Behavioral and electrophysiological evidence for volatile sex pheromones in Parcoblatta wood cockroaches

Species within the cockroach genus Parcoblatta are sexually dimorphic for wing length; females have reduced wings and are flightless, while males have long wings that are used in flight. We predicted that Parcoblatta females would release a volatile sex pheromone to attract the more mobile males. Nymphs of the broad wood cockroach, P. lata, and the Caudell's wood cockroach, P. caudelli, were collected in forested areas in North Carolina, USA, and reared in the laboratory for observations of sexual behavior and for pheromone analysis. After several days of sexual maturation, virgin females of both species exhibited distinct calling behaviors. In females of P. lata, calling commenced 6 days after adult emergence. Under a light–dark photoperiod regime, calling behavior in both species was restricted to the scotophase. Calling consisted of a repeated pattern of raising and lowering the abdomen with occasional exposure of the genital vestibulum. To test whether calling behavior is associated with the release of pheromone, volatiles from calling and noncalling females were collected on Super-Q and tested by electroantennogram (EAG) and behavioral assays. Volatile collections from calling females elicited higher male-specific EAG responses than collections from noncalling females of the same physiological stage. In an olfactometer choice test (Y-tube), males preferred volatiles from calling females over those from noncalling females. To determine the anatomical source of the pheromone, solvent extracts of various body parts were analyzed by EAG. The first through seventh tergites were the only body parts that elicited male-specific EAG responses in both species. In P. lata, the activity of the extract increased from 1- to 7-day-old females, but was lower in mated than in virgin females of the same age. The putative pheromone gland appears to consist of numerous class-3 secretory units, each composed of a secretory cell connected to a cuticular pore via a tubular duct. We conclude that female P. lata and P. caudelli produce sex-specific volatile pheromones that are emitted during calling behavior.