The Role of Ozone-reactive Compounds, Terpenes, and Green Leaf Volatiles (GLVs), in the Orientation of Cotesia plutellae

The emission of inducible volatile organic compounds (VOCs), i.e., inducible terpenes, and green leaf volatiles (GLVs), is a common response of plants to herbivore attack. These VOCs are involved in the orientation of natural enemies, i.e., predators and parasitoids, toward their herbivore prey or hosts (indirect defense of plants). Terpenes and some GLVs are readily oxidized by ozone (O₃), an important oxidant of the low atmosphere and predicted to increase as a result of anthropogenic activity. It has been recently reported that O₃ degradation of terpenes and GLVs does not affect signaling in two selected tritrophic systems. Natural enemies may have learned to use oxidation products that are more stable in nature to locate their prey. To understand the role of these compounds on the tritrophic system Brassica oleracea–Plutella xylostella–Cotesia plutellae, we assessed the preference of wasps to different combinations of cabbage VOCs (intact vs. herbivore-induced and herbivore-induced vs. herbivore-induced VOCs) in the presence or absence of O₃. We found that C. plutellae preferred P. xylostella-damaged plants at 0 and 120 nl l⁻¹ O₃ to intact plants at 0 nl l⁻¹ O₃. However, wasps preferred P. xylostella-damaged plants at 0 nl l⁻¹ to P. xylostella-damaged plants at 120 nl l⁻¹ O₃. The results suggest that compounds other than terpenes and GLVs are crucial for the orientation of the wasps, but terpenes and GLVs contribute to the behaviorally active VOC blend of herbivore-damaged cabbages by increasing their attraction to them. The products resulting from oxidation of terpenes and GLVs do not seem to play a role in the host location process as speculated previously.     

Gustatory Response by the Hymenopteran Parasitoid Cotesia glomerata to a Range of Nectar and Honeydew Sugars

The feeding response of food-deprived Cotesia glomerata to solutions of 14 naturally occurring sugars was determined. Glucose, fructose, sucrose, maltose, erlose, melezitose, trehalose, and stachyose all elicited a feeding response. The sugars differed, however, with respect to the lowest concentration at which they were accepted (acceptance threshold). The parasitoids showed no feeding response when presented with 2 M solutions of galactose, mannose, rhamnose, lactose, raffinose, and melibiose. Sugars from the latter group did not show a deterrent effect when offered to water-deprived parasitoids. When mannose, rhamnose, melibiose, or raffinose were combined with low molar solutions of either fructose or sucrose, sucrose acceptance was affected by mannose and raffinose, whereas no negative interactions were found in mixtures with fructose. Compared to acceptance thresholds reported in other insect systems, the responses of C. glomerata differ considerably with respect to both the range of saccharides accepted as well as the acceptance thresholds. The novel finding that the parasitoid accepts a number of sugars that fail to elicit a feeding response in its herbivorous hosts is of particular interest to the use of (selective) food supplements in biological control programs.     

Herbivore-Induced Volatile Production by Arabidopsis thaliana Leads to Attraction of the Parasitoid Cotesia rubecula: Chemical, Behavioral, and Gene-Expression Analysis

Many plant species defend themselves against herbivorous insects indirectly by producing volatiles in response to herbivory. These volatiles attract carnivorous enemies of the herbivores. Research on the model plant Arabidopsis thaliana (L.) Heynh. has contributed considerably to the unraveling of signal transduction pathways involved in direct plant defense mechanisms against pathogens. Here, we demonstrate that Arabidopsis is also a good candidate for studying signal transduction pathways involved in indirect defense mechanisms by showing that: (1) Adult females of Cotesia rubecula, a specialist parasitic wasp of Pieris rapae caterpillars, are attracted to P. rapae-infested Arabidopsis plants. (2) Arabidopsis infested by P. rapae emits volatiles from several major biosynthetic pathways, including terpenoids and green leaf volatiles. The blends from herbivore-infested and artificially damaged plants are similar. However, differences can be found with respect to a few components of the blend, such as two nitriles and the monoterpene myrcene, that were produced exclusively by caterpillar-infested plants, and methyl salicylate, that was produced in larger amounts by caterpillar-infested plants. (3) Genes from major biosynthetic pathways involved in volatile production are induced by caterpillar feeding. These include AtTPS10, encoding a terpene synthase involved in myrcene production, AtPAL1, encoding phenylalanine ammonia-lyase involved in methyl salicylate production, and AtLOX2 and AtHPL, encoding lipoxygenase and hydroperoxide lyase, respectively, both involved in the production of green leaf volatiles. AtAOS, encoding allene oxide synthase, involved in the production of jasmonic acid, also was induced by herbivory.     

Plant-natural enemy association in tritrophic system,Cotesia rubecula-Pieris rapae-brassicaceae (Cruciferae). III: Collection and identification of plant and frass volatiles

To elucidate the identity of the volatile compounds that could be involved in the searching behavior of the parasitoidCotesia rubecula Marshall (Hymenoptera: Braconidae), the volatiles released by cabbage and frass of Lepidoptera feeding on cabbage were collected and analyzed using a gas chromatograph-mass spectrometer. The volatiles emitted by intact cabbage were -pinene, -pinene, myrcene, 1,8-cineole,n-hexyl acetate,cis-3-hexen-1-yl acetate, and dimethyl trisulfide. Mechanical damage on an intact plant induced the release of two more compounds,trans-2-hexenal and 1-methoxy-3-methylene-2-pentanone. Current feeding by larvae ofPieris rapae L. (Pieridae) induced the plant to release all the compounds released after mechanical damage and additionally 4-methyl-3-pentenal and allyl isothiocyanate. Current feeding by larvae ofPlutella xylostella L. (Plutellidae) induced the plant to release all the compounds present after mechanical damage and additionally allyl isothiocyanate. The volatiles emitted after feeding by the lepidopterans had ceased were the same as those emitted by cabbage damaged by mechanical means. The blend of volatiles emitted by frass was comprised of plant chemicals, mainly sulfur compounds. Frass ofP. rapae emitted allyl isothiocyanate, methyl isothiocyanate, methyl propyl sulfide, dimethyl trisulfide,S-methyl methane thiosulfinate, 4-methyl-3-pentenal,trans-2-hexenal, and 2,3-dihydro-4-methyl furan. Frass ofP. xylostella emitted only dimethyl trisulfide andS-methyl methane thiosulfinate. The blend of volatiles emitted by frass is herbivore-species specific.     

Olfactory Responses of Plutella xylostella Natural Enemies to Host Pheromone, Larval Frass, and Green Leaf Cabbage Volatiles

The parasitoids Trichogramma chilonis (Hymenoptera: Trichogrammatidae) and Cotesia plutellae (Hymenoptera: Braconidae), and the predator Chrysoperla carnea (Neuroptera: Chrysopidae), are potential biological control agents for the diamondback moth, Plutella xylostella (Lepidoptera: Yponomeutidae). We present studies on the interactions between these bioagents and various host-associated volatiles using a Y olfactometer. T. chilonis was attracted to a synthetic pheromone blend (Z11–16:Ald, Z11–16:Ac, and Z11–16:OH in a 1:1:0.01 ratio), to Z11–16:Ac alone, and to a 1:1 blend of Z11–16:Ac and Z11–16:Ald. C. plutellae responded to the blend and to Z11–16:Ac and Z11–16:Ald. Male and female C. carnea responded to the blend and to a 1:1 blend of the major components of the pheromone, although no response was elicited by single compounds. Among the four host larval frass volatiles tested (dipropyl disulfide, dimethyl disulfide, allyl isothiocyanate, and dimethyl trisulfide), only allyl isothiocyanate elicited significant responses in the parasitoids and predator, but C. plutellae and both sexes of C. carnea did respond to all four volatiles. Among the green leaf volatiles of cabbage (Brassica oleracea subsp. capitata), only Z3–6:Ac elicited significant responses from T. chilonis, C. plutellae, and C. carnea, but C. plutellae also responded to E2–6:Ald and Z3–6:OH. When these volatiles were blended with the pheromone, the responses were similar to those elicited by the pheromone alone, except for C. carnea males, which had an increased response. The effect of temperature on the response of the biological agents to a mixture of the pheromone blend and Z3–6:Ac was also studied. T. chilonis was attracted at temperatures of 25–35°C, while C. plutellae and C. carnea responded optimally at 30–35°C and 20–25°C, respectively. These results indicate that the sex pheromone and larval frass volatiles from the diamondback moth, as well as volatile compounds from cabbage, may be used by these natural enemies to locate their diamondback moth host.     

Plant-natural enemy association in the tritrophic system,Cotesia rubecula-Pieris rapae-brassiceae (cruciferae): I. Sources of infochemicals

The role of airborne infochemicals in host selection by the parasitoidCotesia rubecula (Marshal) (Hymenoptera: Braconidae) was examined in a wind tunnel. To elucidate the role of volatile chemicals in attractingC. rubecula to cabbage infested by the host [Pieris rapae L. (Lepidoptera: Pieridae)], the potential sources of volatiles related toP. rapae infestation on cabbage were tested individually. The responses of females to nonhost plant species, bean and geranium, as well as to frass of a nonhost lepidopteran were also examined.C. rubecula was attracted to cabbage previously infested byP. rapae and to frass and regurgitate ofP. rapae. No attraction was observed to larvae ofP. rapae alone. Females were also attracted to mechanically damaged cabbage, cabbage previously infested byPlutella xylostella L. (Lepidoptera: Plutellidae) (a nonhost lepidopteran herbivore), and cabbage previously infested by snails (a nonhost, noninsect herbivore). Intact cabbage, bean, and geranium plants elicited no attraction. A low frequency of attraction was observed to mechanically damaged bean and geranium. Attraction was also observed to frass ofP. xylostella. Volatiles from cabbage related to damage, and volatiles from frass and regurgitate of the host seem to play an important role in guidingC. rubecula to plants infested by its host.     

Learning of Herbivore-Induced and Nonspecific Plant Volatiles by a Parasitoid,Cotesia kariyai

Learning of host-induced plant volatiles by Cotesia kariyai females was examined with synthetic chemicals in a wind tunnel. Wasps were preconditioned by exposure to volatiles and feces simultaneously. A blend of four chemicals, geranyl acetate, -caryophyllene, (E)--farnesene, and indole, which are known to be specifically released from plants infested by host larvae Mythimna separata (host-induced blend), elicited a response in naive C. kariyai, but did not enhance the response after conditioning. A blend of five chemicals, (E)-2-hexenal, (Z)-3-hexen-1-ol, (Z)-3-hexen-1-yl acetate, -myrcene, and linalool, which are known to be released not only from plants infested by the host larvae, but also from artificially damaged plants or undamaged ones (unspecific blend), elicited little response in naive wasps, but significantly enhanced the wasps' response after conditioning. With a blend of the above nine chemicals, wasps could learn the blend at lower concentrations than they did in the nonspecific blend. Hence, both the host-induced and nonspecific volatile compounds appear to be important for C. kariyai females to learn the chemical cues in host location.     

Impact of Botanical Pesticides Derived from Melia azedarach and Azadirachta indica Plants on the Emission of Volatiles that Attract Parasitoids of the Diamondback Moth to Cabbage Plants

Herbivorous and carnivorous arthropods use chemical information from plants during foraging. Aqueous leaf extracts from the syringa tree Melia azedarach and commercial formulations from the neem tree Azadirachta indica, Neemix 4.5^®, were investigated for their impact on the flight response of two parasitoids, Cotesia plutellae and Diadromus collaris. Cotesia plutellae was attracted only to Plutella xylostella-infested cabbage plants in a wind tunnel after an oviposition experience. Female C. plutellae did not distinguish between P. xylostella-infested cabbage plants treated with neem and control P. xylostella-infested plants. However, females preferred infested cabbage plants that had been treated with syringa extract to control infested plants. Syringa extract on filter paper did not attract C. plutellae. This suggests that an interaction between the plant and the syringa extract enhances parasitoid attraction. Diadromus collaris was not attracted to cabbage plants in a wind tunnel and did not distinguish between caterpillar-damaged and undamaged cabbage plants. Headspace analysis revealed 49 compounds in both control cabbage plants and cabbage plants that had been treated with the syringa extract. Among these are alcohols, aldehydes, ketones, esters, terpenoids, sulfides, and an isothiocyanate. Cabbage plants that had been treated with the syringa extract emitted larger quantities of volatiles, and these increased quantities were not derived from the syringa extract. Therefore, the syringa extract seemed to induce the emission of cabbage volatiles. To our knowledge, this is the first example of a plant extract inducing the emission of plant volatiles in another plant. This interesting phenomenon likely explains the preference of C. plutellae parasitoids for cabbage plants that have been treated with syringa extracts.     

Effect of the Presence of a Nonhost Herbivore on the Response of the Aphid Parasitoid <Emphasis Type="Italic">Diaeretiella rapae</Emphasis> to Host-infested Cabbage Plants

The vast majority of studies of plant indirect defense strategies have considered simple tritrophic systems that involve plant responses to attack by a single herbivore species. However, responses by predators and parasitoids to specific, herbivore-induced, volatile blends could be compromised when two or more different herbivores are feeding on the same plant. In Y-tube olfactometer studies, we investigated the responses of an aphid parasitoid, Diaeretiella rapae (McIntosh) (Hymenoptera: Braconidae), to odors from cabbage plants infested with the peach-potato aphid Myzus persicae (Sulzer) (Homoptera: Aphididae), in both the presence and absence of a lepidopteran caterpillar, Plutella xylostella L. (Lepidoptera: Plutellidae). Female parasitoids chose aphid-infested plants over uninfested plants but did not distinguish between caterpillar-infested and uninfested plants. When given a choice between odors from an aphid-infested plant and those from a plant infested with diamondback moth larvae, they significantly chose the former. Furthermore, the parasitoids responded equally to odors from a plant infested with aphids only and those from a plant infested with both aphids and caterpillars. The results support the hypothesis that the aphid and the caterpillar induce different changes in the volatile profile of cabbage plants and that D. rapae females readily distinguish between the two. Furthermore, the changes to the plant volatile profile induced by the caterpillar damage did not hinder the responses of the parasitoid to aphid-induced signals.     

Plant-natural enemy association in the tritrophic systemCotesia rubecula-Pieris rapae-brassicaceae (cruciferae): II. Preference ofC. rubecula for landing and searching

The responses of the parasitoidCotesia rubecula to differently damaged cabbages were recorded during a series of choice tests. To determine if flyingC. rubecula can discriminate differences in the blend of volatiles emitted by cabbages damaged by different causes and how plant volatiles released from a distant source affect the searching behavior ofC. rubecula once searching on a plant, wasps were presented with a choice of plants located one behind the other and separated by a distance of 15 cm. The sources of damage were: cabbage damaged by the host (Pieris rapae), by a nonhost lepidopteran herbivore (Plutella xylostella), by a nonhost, noninsect herbivore (snail), and by mechanical means. The results showed that the site of first landing and the time spent searching on the leaves was influenced by the type of damage inflicted on plants. Wasps preferred to land on cabbages damaged by host and nonhost species of Lepidoptera over those damaged by snails and mechanical means. No preference was observed for first landing between cabbages damaged by the two species of Lepidoptera or between cabbages damaged by snails and mechanical means. Cabbage damaged byP. rapae was searched most intensively, followed by cabbage damaged byP. xylostella, cabbage damaged by snails, and cabbage damaged by mechanical means.C. rubecula differentiates between the volatile blends emitted by differently damaged cabbages, and it is attracted to volatiles related to recent lepidopteran damage. Wasps searched longer on freshly damaged than on leaves with older damage.     

Induction of parasitoid attracting synomone in brussels sprouts plants by feeding ofPieris brassicae larvae: Role of mechanical damage and herbivore elicitor

Induction of plant defense in response to herbivory includes the emission of synomones that attract the natural enemies of herbivores. We investigated whether mechanical damage to Brussels sprouts leaves (Brassica oleracea var.gemmifera) is sufficient to obtain attraction of the parasitoidCotesia glomerata or whether feeding byPieris brassicae caterpillars elicits the release of synomones not produced by mechanically damaged leaves. The response of the parasitoidCotesia glomerata to different types of simulated herbivory was observed. Flight-chamber dual-choice tests showed that mechanically damaged cabbage leaves were less attractive than herbivore-damaged leaves and mechanically damaged leaves treated with larval regurgitant. Chemical analysis of the headspace of undamaged, artificially damaged, caterpillar-infested, and caterpillar regurgitant-treated leaves showed that the plant responds to damage with an increased release of volatiles. Greenleaf volatiles and several terpenoids are the major components of cabbage leaf headspace. Terpenoids are emitted in analogous amounts in all treatments, including undamaged leaves. On the other hand, if the plant is infested by caterpillars or if caterpillar regurgitant is applied to damaged leaves, the emission of green-leaf volatiles is highly enhanced. Our data are in contrast with the induction of more specific synomones in other plant species, such as Lima bean and corn.     

Response of the Aphid Parasitoid <Emphasis Type="Italic">Aphidius funebris</Emphasis> to Volatiles from Undamaged and Aphid-infested <Emphasis Type="Italic">Centaurea nigra</Emphasis>

Two issues have hindered the understanding of the ecology and evolution of volatile-mediated tritrophic interactions: few studies have addressed noncrop systems; and few statistical techniques have been applied that are suitable for the analysis of complex volatile blends. In this paper, we addressed both of these issues by studying the noncrop system involving the plant Centaurea nigra, the specialist aphid Uroleucon jaceae, and the parasitoid Aphidius funebris. In a Y-tube olfactometer, A. funebris was attracted to the odor from undamaged C. nigra, but preferred the plant–host complex (PHC) after 3 d of feeding by 200 U. jaceae over the undamaged plant, but not after three or 5 d of feeding by 50 U. jaceae. When aphids were removed, the initial preference for the damaged plant remained, but the final preference was not greater than for the undamaged plant. No qualitative differences were detected between the headspaces of C. nigra and the C. nigra–U. jaceae PHC. For quantitative analysis, we used a compositional approach, which treats each compound produced as part of a blend, and not as a compound released in isolation, thus allowing analysis of the relative contribution of each compound to the blend as a whole. With this approach, subtle increases and decreases of some green leaf volatiles and monoterpenoids on the third day of aphid infestation were detected. Mechanically damaged C. nigra had a volatile profile that differed from undamaged C. nigra and the PHC. One and 10 ng of (Z)-3-hexenyl acetate, and 10 or 100 ng of 6-methyl-5-hepten-2-one were attractive to the parasitoid when placed in solution on filter paper. A. funebris appears to be using a combination of chemical cues to locate host-infested plants.     

Selection of Nothofagus Host Trees by the Aphids Neuquenaphis staryi and Neuquenaphis edwardsi

Leaf volatiles were collected from three Nothofagus species growing in close proximity in Los Ruiles National Reserve, Chile. The volatile preparation from leaves of No. alessandrii were attractive to the specialist aphid, Neuquenaphis staryi, but not to the generalist aphid, Ne. edwardsi, while the volatile preparations of No. dombeyi and No. glauca were attractive to Ne. edwardsi, but not to Ne. staryi. This reflects the pattern of aphid/host-plant associations. Alpha-Agarofuran was found to occur in all leaf volatile preparations and was shown by electroantennography and olfactometry to be attractive for both Neuquenaphis spp., suggesting it may be the Nothofagus host-recognition factor for Neuquenaphis. The factor(s) mediating Ne. stayi's specialization on No. alessandrii remain to be identified.     

The Role of Indole and Other Shikimic Acid Derived Maize Volatiles in the Attraction of Two Parasitic Wasps

After herbivore attack, plants release a plethora of different volatile organic compounds (VOCs), which results in odor blends that are attractive to predators and parasitoids of these herbivores. VOCs in the odor blends emitted by maize plants (Zea mays) infested by lepidopteran larvae are well characterized. They are derived from at least three different biochemical pathways, but the relative importance of each pathway for the production of VOCs that attract parasitic wasps is unknown. Here, we studied the importance of shikimic acid derived VOCs for the attraction of females of the parasitoids Cotesia marginiventris and Microplitis rufiventris. By incubating caterpillar-infested maize plants in glyphosate, an inhibitor of the 5-enolpyruvylshikimate-3-phospate (EPSP) synthase, we obtained induced odor blends with only minute amounts of shikimic acid derived VOCs. In olfactometer bioassays, the inhibited plants were as attractive to naive C. marginiventris females as control plants that released normal amounts of shikimic acid derived VOCs, whereas naive M. rufiventris females preferred inhibited plants to control plants. By adding back synthetic indole, the quantitatively most important shikimic acid derived VOC in induced maize odors, to inhibited plants, we showed that indole had no effect on the attraction of C. marginiventris and that M. rufiventris preferred blends without synthetic indole. Exposing C. marginiventris females either to odor blends of inhibited or control plants during oviposition experiences shifted their preference in subsequent olfactometer tests in favor of the experienced odor. Further learning experiments with synthetic indole showed that C. marginiventris can learn to respond to this compound, but that this does not affect its choices between natural induced blends with or without indole. We hypothesize that for naïve wasps the attractiveness of an herbivore-induced odor blend is reduced due to masking by nonattractive compounds, and that during oviposition experiences in the presence of complex odor blends, parasitoids strongly associate some compounds, whereas others are largely ignored.     

Chemical Composition of Corn Leaf Essential Oils and Their Role in the Oviposition Behavior of Sesamia nonagrioides Females

The chemical composition of the volatile oils collected by steam distillation from leaves of seven corn hybrids and their effect on the oviposition behavior of Sesamia nonagrioides females was studied. Samples of the volatile oils from each of the hybrids were analyzed by capillary gas chromatography-mass spectrometry (GC-MS) and several major compounds were identified. The major compound found in all seven hybrids was 3,7,11,15-tetramethyl-2-hexadecen-1-ol (phytol), ranging from 38.3 to 64.9% of the total quantity. Compounds detected in significant proportions include (Z)-3-hexenol (3.1 to 8%), nonanal (4.9 to 14.5%), pentadecanal (1.8 to 5.8%), neophytadiene (5.5 to 12.9%), (Z)-3-hexenyl acetate (2.5 to 8.9%), and an analogue of 2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3-(4H)-one (DIMBOA) (2.4 to 9.3%). The analysis showed no qualitative differences among the chemicals identified while quantitative differences were detected. Among the volatile oils, the significant difference was in the quantity of aldehydes present. In two-choice bioassays, filter paper sticks treated with volatile oils containing higher quantity of aldehydes received fewer eggs than those with lower aldehyde quantity. Bioassays with synthetic aldehydes of a chain length C9-C14 confirmed the above results.     

Influence of the Type of Reducing Agent (H2, CO, C3H6 and C3H8) on the Reduction of Stored NOX in a Pt/BaO/Al2O3 Model Catalyst

In this investigation, a comparative study for a NO _X storage catalytic system was performed focusing on the parameters that affect the reduction by using different reductants (H₂, CO, C₃H₆ and C₃H₈) and different temperatures (350, 250 and 150 °C), for a Pt/BaO/Al₂O₃ catalyst. Transient experiments show that H₂ and CO are highly efficient reductants compared to C₃H₆ which is somewhat less efficient. H₂ shows a significant reduction effect at relatively low temperature (150 °C) but with a low storage capacity. We find that C₃H₈does not show any NO _X reduction ability for NO _X stored in Pt/BaO/Al₂O₃ at any of the temperatures. The formation of ammonia and nitrous oxide is also discussed.