Evidence for Additional Pheromonal Components Mediating Oviposition Aggregation in Schistocerca gregaria

Oviposition responses of gregarious adult gravid females of the desert locust Schistocerca gregaria to sand previously used for oviposition by gregarious females (contaminated sand, with eggs and froth removed) and to sterilized sand (noncontaminated sand) were compared in bioassays. Gregarious females preferentially oviposited into contaminated sand. The number of egg pods laid by the females was found to correlate with the frequency of use of the contaminated sand for oviposition. Charcoal-trapped volatiles from the contaminated sand also elicited similar responses from gregarious females. Gas chromatography–electroantennographic detection (GC-EAD) analysis of the trapped volatiles revealed the presence of three electrophysiologically active compounds that were identified by GC-MS as (Z)-6-octen-2-one, (E,E)-3,5-octadien-2-one and its geometric isomer (E,Z)-3,5-octadien-2-one. The relative amounts of the three EAG-active compounds in the volatiles increased with consecutive oviposition by the gregarious females. These results indicate mediation of additional pheromonal components in the oviposition behavior of gregarious desert locust that are distinct from those that were previously identified in the froth volatiles.     

The nature of the gregarizing signal responsible for maternal transfer of phase to the offspring in the desert locust Schistocerca gregaria

We examined aggregative behavior of hatchlings of the desert locust Schistocerca gregaria emerging from solitaria egg pods that had been incubated in sand previously used for consecutive ovipositions by gregarious females. Hatchlings derived from such eggs were significantly more gregarious than those derived from pods laid in clean sand. The gregarizing effect of the sand-associated factor originating from 3, 5, or 10 ovipositions by gregarious females increased in a dose-dependent fashion. Washing the sand with organic solvents following such ovipositions, or flushing it with nitrogen gas, led to substantial loss of its gregarizing effect, showing that the active signal is volatile and of medium polarity. The gregarizing activity of the exposed sand correlated with the presence of C-8 unsaturated ketones, (Z)-6-octen-3-one and (E,E)-3,5-octadiene-2-one and its E,Z isomer, previously shown by us to form part of the releaser pheromone system that mediates group oviposition in S. gregaria. These ketones were present in relatively large amounts in the eggs obtained from egg pods of gregarious females and were also detected in the extracts of accessory glands of gregarious females, a candidate source of the gregarization factor. It is proposed that the pheromone is secreted at the onset of oviposition. This would account for its distribution predominantly within the eggs and surrounding sand at the site of oviposition. The study sheds new light on the pheromonal mechanism associated with transgenerational transmission of gregarious characters in crowded S. gregaria populations and provides yet another case in this insect of dual releaser and primer roles played by the same pheromone blend.     

Cuticular Surface Hydrocarbons of Desert Locust Nymphs, Schistocerca gregaria, and Their Effect on Phase Behavior

The quantity of cuticular hydrocarbons is higher in solitarious nymphs of the desert locust, Schistocerca gregaria, compared to gregarious nymphs, but the total hydrocarbon fraction of solitarious nymphs does not significantly divert behavioral transition of isolated nymphs to the gregarious phase, while gregarious hydrocarbon extracts do. This suggests that qualitative differences in composition are responsible for the biological effect. The profile of cuticular hydrocarbon components is similar in the two phases, but some peak ratios differ. Crowding of solitarious nymphs leads to rapid changes in the profile of the hydrocarbon fraction, suggesting that specific hydrocarbons are produced and secreted as a consequence. Isolating previously crowded nymphs has an opposite effect. The composition of cuticular hydrocarbons from the migratory locust, Locusta migratoria, which differs considerably from that of S. gregaria, does not induce the gregarious behavioral phase in solitarious nymphs of the latter.     

What defense doesSchouwia purpurea (Cruciferae) have against the desert locust?

A field study of the food eaten by solitary desert locusts was carried out in a winter breeding area in Mauritania. The food eaten, determined by the plant epidermis found in fecal pellets, was compared to the plant's availability in the habitat.Schouwia purpurea, well represented in the diets, was dominant at the study site. Adults had a preference forTribulus terrester. Growth and feeding on these two plants were compared. The high water content ofS. purpurea leaves limited the dry matter eaten and slowed down growth. Glucosinolates were separated and quantified by gas chromatography. There are 132mol/g dry matter in green leaves. In multiple choice tests, with paper disks, glucosinolate extracts were phagostimulant at a low concentration (21mol/g dry matter) and repulsive at a higher one (214mol/ g dry matter). Biting behavior onS. purpurea was recorded and analyzed on video. The importance ofSchouwia purpurea in desert locust habitats and its defenses is discussed.     

Sex pheromone of Egyptian cotton leafworm (Spodoptera littoralis)

Samples of pheromone carriers, after use for 4–5 weeks in traps in the field, were analyzed. Partial isomerization of prodlure, (Z,E)-9,11-tetradecadienyl acetate (TDDA), the main component of the sex pheromone of femaleSpodoptera littoralis, took place. All three other possible isomers were formed to yield a mixture of 50% (Z,E)-9,11 -TDDA, 30% (E,E)-9,11-, 13% (E,Z)-9,11-, and 7% (Z,Z)-9,11 isomer. The process was found to be photochemically induced; none of these isomers was found under heating alone. Most of the pheromone, however, decomposed under field conditions.     

Factors affecting behavioral phase transition in the desert locust,Schistocerca gregaria (Forskål) (Orthoptera: Acrididae)

The behavior of the desert locust,Schistocerca gregaria (Forskål) (Orthoptera: Acrididae), is adjusted rapidly to population density and is a phase characteristic. We used discriminant analysis to quantify the extent of phase transition from the solitary to the gregarious phase and accurately classify the phase status on the basis of two decisive behavioral parameters: nymphal activity and social interaction. Fecal extracts. examined by olfactometry, attract solitarious nymphs but do not contribute to behavioral phase transition. Neither do visual stimuli alone. Short-range olfaction of airborne volatiles slightly affects behavioral phase transition. Antennectomy abolishes density response. Cuticular lipid extract, containing presumptive contact pheromones, does not attract nymphs, but does significantly affect behavioral phase transition.     

Evidence for mediation of two releaser pheromones in the aggregation behavior of the gregarious desert locust,Schistocerca gregaria (forskal) (Orthoptera: Acrididae)

The response of nymphal and adult gregarious phase desert locust,Schistocerca gregaria, to a choice of two columns of air, one permeated with airborne volatiles emanating from nymphs or adults and the other untreated, was investigated in a single-chamber bioassay arena. The nymphs, whether released individually or in groups, preferred to be within the precinct of the air column treated with airborne volatiles of the nymphs but were indifferent to volatiles of the adults. Conversely, older adults responded only to their own volatiles but not to those of the nymphs or young adults. The young adults were responsive only to volatiles of the older adults. Charcoal-trapped volatiles from the nymphs and the adults reproduced the effect of living locusts. These results indicate that there are two different aggregation pheromones inS. gregaria: a juvenile pheromone produced by nymphs and an adult pheromone specific to adults.     

Oviposition and Behavioral Responses of Culex quinquefasciatus to Skatole and Synthetic Oviposition Pheromone in Tanzania

Studies on the behavioral and oviposition responses of Culex quinquefasciatus to 3-methylindole or skatole (an oviposition site cue) and (5R,6S)-6-acetoxy-5-hexadecanolide (the synthetic oviposition pheromone, SOP) were conducted in Muheza, northeast Tanzania. Cx. quinquefasciatus displayed a bimodal pattern of oviposition. The highest peak (64.8%) was between 18:00 and 20:00 hr, while the second peak (18.9%) was between 06:00 and 08:00 hr. The mean number of egg rafts oviposited per day between the two peaks was different (P < 0.05). Oviposition responses of gravid Cx. quinquefasciatus differed across the range of concentrations of skatole. Larger numbers of egg rafts were deposited in bowls treated with 10^–6 g or 10^–5 g skatole than in bowls with tap water (P < 0.05). The number of egg rafts deposited in bowls treated with 10^–4 or 10^–1 g skatole was not different from that laid in tap water (P > 0.05). Under field conditions skatole was found to be effective for a period of seven days, after which its biological activity decreased significantly. It was further found that more egg rafts of Cx. quinquefasciatus were deposited in water treated with SOP than in water treated with skatole (P = 0.0002). An additive effect was observed when the SOP + skatole combination was used to attract gravid Cx. quinquefasciatus to oviposition sites. It is concluded that skatole and SOP mediate oviposition site selection of Cx. quinquefasciatus under field conditions.     

EFFECT OF PBAN ON PHEROMONE PRODUCTION BY MATED <Emphasis Type="Italic">Heliothis virescens</Emphasis> AND <Emphasis Type="Italic">Heliothis subflexa</Emphasis> FEMALES

Mated female Heliothis virescens and H. subflexa were induced to produce sex pheromone during the photophase by injection of pheromone biosynthesis activating neuropeptide (PBAN). When injected with 1 pmol Hez-PBAN, the total amount of pheromone that could be extracted from glands of mated females during the photophase was similar to that extracted from virgin females in the scotophase. The PBAN-induced profile of pheromone components was compared between mated, PBAN-injected females and virgin females during spring and fall. Virgin females exhibited some differences in the relative composition of the pheromone blend between spring and fall, but no such temporal differences were detected in PBAN-injected, mated females. Because the temporal variation in pheromone blend composition was greater for virgin females than for PBAN-injected females, PBAN can be used to determine a females native pheromone phenotype. This procedure has the advantages that pheromone glands can be extracted during the photophase, from mated females that have already oviposited.     

Identification of female sex pheromone in alfalfa blotch leafminer,Agromyza frontella (Rondani)(Diptera: Agromyzidae)

Gas chroinatography of hexane extracts of 3-day-old virgin A.frontella adults revealed a branched saturated hydrocarbon present only in females. This compound was isolated by argentation chromotography and preparative GC, identified by GC and GC-MS as 3,7-dimethylnonadecane, and subsequently synthesized. Three-day-old virgin females contained 54.1 ± 3.5 ng (± SEM;N = 48) of 3,7-dimethylnonadecane. Male cadavers do not elicit male courtship behavior, but when treated with 18 ng of 3,7-dimethylnonadecane they were as attractive as cadavers of 3-day-old virgin females. Black cotton knots were less attractive than male cadavers when treated with 3,7-dimethylnonadecane, but all bioassays indicated that this compound is an important semiochemical modulating male mating behavior.     

Composition and role of volatile substances in atmosphere surrounding two gregarious locusts,Locusta migratoria andSchistocerca gregaria

Volatile substances in the atmosphere surrounding gregarious locustsSchistocerca gregaria andLocusta migratoria were captured and investigated by combined gas chromatography-mass spectrometry. Three aromatic derivates have been identified: phenol, guaiacol, and veratrole. Their relative percentages differ for different ages and species. Behavioral tests show that essentially phenol, guaiacol, and the mixture of the three products tend to increase the aggregation behavior in both species and thus act as cohesion pheromones.     

Variability in Flavonoid Compounds of Four Tribulus Species: Does it Play a Role in Their Identification by Desert Locust Schistocerca gregaria?

We examined how variability in phenolic metabolism of four closely related Tribulus taxa might affect palatability to Schistocerca gregaria. Plants were collected in the Mauritanian breeding areas of the locust. Eighteen flavonoids (caffeoyl derivatives, quercetin glycosides, including rutin, and kaempferol glycosides) were detected with high-performance liquid chromatography in leaf extracts. Principal component analysis (PCA) differentiated relative abundance of flavonoids of Tribulus terrestris from T. longipetalus, T. ochroleucus, and T. bimucronatus. Flavonoid content of T. terrestris leaves varied between northern and southern Mauritanian sites. Dual choice tests with plants whose flavonoid profiles had been determined previously were analyzed by multiple regression. The data suggest that the desert locust prefers T. terrestris plants rich in quercetin glycosides.     

Sex pheromone communication in the nematode,Rhabditis pellio

Both males and females ofRhabditis pellio release pheromones that attract the opposite sex prior to copulation. A quantitative bioassay for the female-produced pheromone was designed, based on male movement toward a pheromone source placed at one end of a 10-mm strip of bacterial material maintained on nutrient agar in a petri plate. Females produced pheromone from the age at which they attained the adult stage (3 days following hatching from the egg) and maintained a relatively constant production level until at least the ninth day of life. Similarly, males became responsive to the female pheromone by the third day and remained responsive through the ninth day, although the time required for the males to migrate toward a female pheromone source increased with increasing age. No daily rhythm of pheromone responsiveness by males or pheromone production by females was observed when the nematodes were conditioned to a 1212 h light-dark cycle.     

Nerol: An alarm substance of the stingless bee,Trigona fulviventris (Hymenoptera: Apidae)

Bees of the genusTrigona and subgenusTrigona possess volatile materials in their mandibular glands, used as alarm substances and as marking pheromones. Heads of workers ofTrigona fulviventris were analyzed by gas chromatography-mass spectrometry. The two major volatile components were nerol ( 50%), and octyl caproate ( 20%). Relative to other substances tested at a Costa Rican nest, treatments containing 20 g of nerol attractedT. fulviventris, depressed numbers of bees leaving the nest by about 50%, and elicited wing vibration and biting. The responses were similar to those obtained with the contents of one worker head. Attraction and biting were also seen in response to captures of colony members by assassin bugs (Apiomerus pictipes) outside a nest entrance; one bee responded in about 15% of the captures. This alarm behavior, although weak, is of interest since it was thought thatT. fulviventris was unusual for its subgenus in its lack of nest defense behaviors.     

The Desert Locust, <Emphasis Type="Italic">Schistocerca gregaria</Emphasis>, Detoxifies the Glucosinolates of <Emphasis Type="Italic">Schouwia purpurea</Emphasis> by Desulfation

Desert locusts (Schistocerca gregaria) occasionally feed on Schouwia purpurea, a plant that contains tenfold higher levels of glucosinolates than most other Brassicaceae. Whereas this unusually high level of glucosinolates is expected to be toxic and/or deterrent to most insects, locusts feed on the plant with no apparent ill effects. In this paper, we demonstrate that the desert locust, like larvae of the diamondback moth (Plutella xylostella), possesses a glucosinolate sulfatase in the gut that hydrolyzes glucosinolates to their corresponding desulfonated forms. These are no longer susceptible to cleavage by myrosinase, thus eliminating the formation of toxic glucosinolate hydrolysis products. Sulfatase is found throughout the desert locust gut and can catalyze the hydrolysis of all of the glucosinolates present in S. purpurea. The enzyme was detected in all larval stages of locusts as well as in both male and female adults feeding on this plant species. Glucosinolate sulfatase activity is induced tenfold when locusts are fed S. purpurea after being maintained on a glucosinolate-free diet, and activity declines when glucosinolates are removed from the locust diet. A detoxification system that is sensitive to the dietary levels of a plant toxin may minimize the physiological costs of toxin processing, especially for a generalist insect herbivore that encounters large variations in plant defense metabolites while feeding on different species.     

Chemistry of Leichhardt's Grasshopper, Petasida ephippigera, and its Host Plants, Pityrodia jamesii, P. ternifolia, and P. pungens

Investigation of the strikingly aposematic Leichhardt's grasshopper revealed a number of host-plant-derived components. Chemical study of the three known host plants revealed a diverse array of components. Pityrodia ternifolia and P. pungens contain predominantly sesquiterpenes and sesquiterpene glycosides and P. jamesii is rich in cineole, dihydrochalcones, chalcones, and monoterpene glycosides. Feces from grasshoppers feeding on P. jamesii contain a mixture of hydroxycineoles, of which 3-hydroxycineole is the predominant isomer (68%). No evidence for previously suspected "alkaloidal" toxins was found in either the insect or host plant.     

Floral volatiles ofTanacetum vulgare L. attractive toLobesia botrana den. et schiff. females

The European grapevine moth (EGVM),Lobesia botrana, is a major pest of grapes in Europe. Females are attracted to a nonhost plant: tansy (Tanacetum vulgare L.), which is a common weed in Slovakian vineyards. A steam distillate extract of tansy flowers was analyzed by means of a GC-EAG technique to screen constituents detected by the olfactory receptors of EGVM females. From more than 200 GC peaks, nine peaks corresponding to monoterpenoids released an EAG response in more than 70% of the females (N=15):p-cymene,d-limonene,-thujene,-thujone,-thujone, thujyl alcohol, terpinene-4-ol, (Z)-verbenol, and piperitone. The steam distillate of tansy as well as a synthetic blend of identified compounds released consistent attraction in a field cage. The use of nonhost plants and host plant odors in integrated pest management is discussed.     

Selectively fluorinated analogs reveal differential olfactory reception and inactivation of green leaf volatiles in insects

The role of the alkyl terminus of green leaf volatile (GLV) molecules in olfactory reception and inactivation was examined in three diverse insect species: the beet armyworm,Spodoptera exigua (Lepidoptera); the Colorado potato beetle,Leptinotarsa decemlineata (Coleoptera); and the desert locust,Schistocerca gregaria (Orthoptera), using selectively fluorinated analogs of GLVs and electroantennograms (EAGs). When only the magnitude of the depolarization of the EAG is considered (a measure of reception), the order of effectiveness was 1-hexanol (6:OH)=(Z)-3-6:OH > 5,5,6,6,6-pentafluoro-(Z)-3-6:OH =5,5-difluoro-(Z)-3-6:OH 5,5,6,6,6-pentafluoro-6: OH. Percent recovery of the EAG (a measure of inactivation) was greater for the pentafluoro-(Z)-3-6: OH analog than for the difluoro-(Z)-3-6: OH analog. Our results show that the alkyl end of GLV molecules plays an important role not only in reception, but also inactivation processes in insect olfaction. Furthermore, specificities of these two processes may differ.Research conducted by J.C.D. while on an ARS Fellowship at IPO-DLO.     

A flavonoid glucoside,phellamurin, regulates differential oviposition on a rutaceous plant,Phellodendron amurense,by two sympatric swallowtail butterflies,Papilio protenor andP. xuthus: The front line of a coevolutionary arms race?

We studied the chemical basis for the differential acceptance of a rutaceous plant.Phellodendron amurense, by ovipositing females of two sympatricPapilio butterflies,P. protenor andP. xuthus, that have a very similar host range.P. amurense, on which larvae of both species perform well, was rejected byP. protenor females but was marginally accepted byP. xuthus, even though water-soluble fraction of this plant elicited significant ovipositional responses from the two species. A prenylated dihydroflavonol glucoside, 3,5,7,4-tetrahydroxy-8-(3-methylbut-2-enyl)flavanone-7-O--glucoside (phellamurin), was identified as a dominant active substance that deters oviposition.P. protenor responded highly sensitively to phellamurin, whereasP. xuthus was less susceptible and was affected conspicuously only at higher concentrations than that found in living plants, thereby accounting for their differential responses toP. amurense. InP. protenor, the deterrent effect of phellamurin was considered to outweigh the activity of coexisting stimulant(s) at the concentrations actually present in the foliage, resulting in avoidance of this plant. The phellamurin content in the foliage amounted to as much as 1.8% of the fresh weight, which is far higher than the average levels of flavonoids present in other major host plants ofPapilio. The dose-response experiments demonstrated that unacceptability ofP. amurense forP. protenor was attributable largely to such a high concentration of phellamurin. These results appear to provide information on possible coevolutionary interactions between herbivores and plants and also on the potential for shifts in host affiliation.     

Sex pheromone of a conifer-feeding budworm,Choristoneura retiniana,Walsingham

Sex pheromone components collected from femaleChristoneura retiniana included 11-tetradecenyl acetates and alcohols. The major component wasE-11-tetradecenyl acetate (E11–14Ac) with a lesser amount ofZ isomer necessary to induce male response. A 928 ratio ofE,Z11–14 Ac appeared optimal. The alcohol component was present at about 10% of the total pheromone mixture, and traps baited with acetates plus alcohol surpassed unmated females in their degree of attractiveness. Chemical analysis indicated a 9:1 ratio of theE — Z isomers of 11-tetradecenyl alcohol (11–14ol) pheromone components, although bait formulations containing a predominance of either theE orZ isomers were equally successful in field bioassays. Based on male response to traps,E- andZ11–14Ac (928E — Z) are essential pheromone components for long-range sex attraction. The 11–14 ol enhanced attraction when added at 10% of the total pheromone blend.This paper reports the results of research only. Mention of a commercial product does not constitute a recommendation by the U.S. Department of Agriculture.