Identification and Field Evaluation of Non-Host Volatiles Disturbing Host Location by the Tea Geometrid, Ectropis obliqua

Volatile organic compounds derived from non-host plants, Ocimum basilicum, Rosmarinus officinalis, Corymbia citriodora, and Ruta graveolens, can be used to mask host plant odors, and are repellent to the tea geometrid, Ectropis obliqua. Volatile compounds were collected by headspace absorption, and the components were identified and quantified by using gas chromatography/mass spectrometry. The responses of antennae of female E. obliqua to the compounds were evaluated with gas chromatography/electroantennography detection. Qualitative and quantitative differences were found among the four odor profiles. Consistent electroantennographic activity was obtained for eight of the volatiles from the four plants: β-myrcene, α-terpinene, γ-terpinene, linalool, cis-verbenol, camphor, α-terpineol, and verbenone. In a Y-tube bioassay, six chemicals, β-myrcene, γ-terpinene, (R)-(?)-linalool, (S)-(?)-cis-verbenol, (R)-(+)-camphor, and (S)-(?)-verbenone, were the main compounds responsible for repelling E. obliqua. An eight-component mixture including all of the bioactive compounds (in a ratio of 13:2:13:8:1:24:6:17) from R. officinalis was significantly more effective at repelling the moths than any single compound or a mixture of equal amounts of the eight compounds. Field results demonstrated that intercropping tea plants with R. officinalis effectively suppressed E. obliqua infestations in a tea plantation. Our findings suggests that odor blends of R. officinalis play a role in disturbing host orientation behavior, and in repelling E. obliqua adults, and that R. officinalis should be considered when developing “push-pull” strategies aimed at optimizing the control of E. obliqua with semiochemicals.     

Specificity of Herbivore-Induced Hormonal Signaling and Defensive Traits in Five Closely Related Milkweeds (Asclepias spp.)

Despite the recognition that phytohormonal signaling mediates induced responses to herbivory, we still have little understanding of how such signaling varies among closely related species and may generate herbivore-specific induced responses. We studied closely related milkweeds (Asclepias) to link: 1) plant damage by two specialist chewing herbivores (milkweed leaf beetles Labidomera clivicolis and monarch caterpillars Danaus plexippus); 2) production of the phytohormones jasmonic acid (JA), salicylic acid (SA), and abscisic acid (ABA); 3) induction of defensive cardenolides and latex; and 4) impacts on Danaus caterpillars. We first show that A. syriaca exhibits induced resistance following monarch herbivory (i.e., reduced monarch growth on previously damaged plants), while the defensively dissimilar A. tuberosa does not. We next worked with a broader group of five Asclepias, including these two species, that are highly divergent in defensive traits yet from the same clade. Three of the five species showed herbivore-induced changes in cardenolides, while induced latex was found in four species. Among the phytohormones, JA and ABA showed specific responses (although they generally increased) to insect species and among the plant species. In contrast, SA responses were consistent among plant and herbivore species, showing a decline following herbivore attack. Jasmonic acid showed a positive quantitative relationship only with latex, and this was strongest in plants damaged by D. plexippus. Although phytohormones showed qualitative tradeoffs (i.e., treatments that enhanced JA reduced SA), the few significant individual plant-level correlations among hormones were positive, and these were strongest between JA and ABA in monarch damaged plants. We conclude that: 1) latex exudation is positively associated with endogenous JA levels, even among low-latex species; 2) correlations among milkweed hormones are generally positive, although herbivore damage induces a divergence (tradeoff) between JA and SA; 3) induction of cardenolides and latex are not necessarily physiologically linked; and 4) even very closely related species show highly divergent induction, with some species showing strong defenses, hormonally-mediated induction, and impacts on herbivores, while other milkweed species apparently use alternative strategies to cope with insect attack.     

Highly Site-Selective Epoxidation of Polyene Catalyzed by Metal–Organic Frameworks Assisted by Polyoxometalate

Ectropis obliqua Prout is one of the most severe defoliator insects of tea plants. Sex pheromones method has been widely used for monitoring and controlling the insect pest of E. obliqua Prout due to their relative safety, low cost, and environmental friendliness. However, conventional preparing sex pheromones of E. obliqua Prout has been criticized by the costly and/or low efficiency procedure. Herein, we report a series of Fe-based metal–organic frameworks (MOFs) as efficient heterogeneous catalysts for the site-selective epoxidation of 3Z,6Z,9Z-octadecatriene to its monoepoxides. Particularly, MIL-100(Fe) combined with polyoxometalate (POM) as cocatalyst appears as the best catalyst for the oxidation of 3Z,6Z,9Z-octadecatriene to a high site selectivity to the corresponding 6,7-epoxide with good yields. 3Z,6Z,9Z-octadecatriene and its corresponding 6,7-epoxide are the primary sex pheromone components of the E. obliqua Prout. Significantly, the field bioassay experiments showed that the mixed products were highly effective in attracting E. obliqua Prout male moths. This work provides a general method to fast and clean synthesis of sex pheromones and opens new avenues for the application of Fe-based MOFs in the pest control with environmental friendliness.     

Body Odors of Parasitized Caterpillars Give Away the Presence of Parasitoid Larvae to Their Primary Hyperparasitoid Enemies

Foraging success of parasitoids depends on the utilization of reliable information on the presence of their often, inconspicuous hosts. These parasitic wasps use herbivore-induced plant volatiles (HIPVs) that provide reliable cues on host presence. However, host searching of hyperparasitoids, a group of parasitoids that parasitize the larvae and pupae of other parasitoids, is more constrained. Their hosts do not feed on plants, and often are even concealed inside the body of the herbivore host. Hyperparasitoids recently have been found to use HIPVs of plants damaged by herbivore hosts in which the parasitoid larvae develop. However, hyperparasitoids that search for these parasitoid larvae may be confronted with healthy and parasitized caterpillars on the same plant, further complicating their host location. In this study, we addressed whether the primary hyperparasitoid Baryscapus galactopus uses caterpillar body odors to discriminate between unparasitized herbivores and herbivores carrying larvae of parasitoid hosts. We show that the hyperparasitoids made faster first contact and spent a longer mounting time with parasitized caterpillars. Moreover, although the three parasitoid hosts conferred different fitness values for the development of B. galactopus, the hyperparasitoids showed similar behavioral responses to caterpillar hosts carrying different primary parasitoid hosts. In addition, a two-chamber olfactometer assay revealed that volatiles emitted by parasitized caterpillars were more attractive to the hyperparasitoids than those emitted by unparasitized caterpillars. Analysis of volatiles revealed that body odors of parasitized caterpillars differ from unparasitized caterpillars, allowing the hyperparasitoids to detect their parasitoid host.     

Induction of volatile emissions in maize by different larval instars of Spodoptera littoralis

Maize plants under attack by caterpillars emit a specific blend of volatiles that is highly attractive to parasitic wasps. The release of these signals is induced by elicitors in the caterpillar regurgitant. Studies suggest that plants respond differently to different herbivore species and even to different herbivore stages, thus providing parasitoids and predators with specific signals. We tested if this is the case for different larval instars of the noctuid moth Spodoptera littoralis when they feed on maize plants. Cut maize plants were incubated in diluted regurgitant from second, third, or fifth instar caterpillars. There were no differences in total amount released after these treatments, but there were small differences in the release of the minor compounds phenethyl acetate and -humulene. Regurgitant of all three instars contained the elicitor volicitin. To test the effect of actual feeding by the larvae, potted plants were infested with caterpillars of one of the three instars, and volatiles were collected the following day. The intensity of the emissions was correlated with the number of larvae feeding on a plant, and with the amount of damage inflicted, but was independent of the instar that caused the damage. We also used artificial damage to mimic the manner of feeding of each instar to test the importance of physical aspects of damages for the odor emission. The emission was highly variable, but no differences were found among the different types of damage. In olfactometer tests, Microplitis rufiventris, a parasitoid that can only successfully parasitize second and early third instar S. littoralis, did not differentiate among the odors of maize plants attacked by different instar larvae. The odor analyses as well as the parasitoid's responses indicate that maize odors induced by S. littoralis provide parasitoids with poor information on the larval developmental stage. We discuss the results in the context of variability and lack of specificity in odorous plant signals.     

Silencing an E3 Ubiquitin Ligase Gene OsJMJ715 Enhances the Resistance of Rice to a Piercing-Sucking Herbivore by Activating ABA and JA Signaling Pathways

The RING-type E3 ubiquitin ligases play an important role in plant growth, development, and defense responses to abiotic stresses and pathogens. However, their roles in the resistance of plants to herbivorous insects remain largely unknown. In this study, we isolated the rice gene OsJMJ715, which encodes a RING-domain containing protein, and investigated its role in rice resistance to brown planthopper (BPH, Nilaparvata lugens). OsJMJ715 is a nucleus-localized E3 ligase whose mRNA levels were upregulated by the infestation of gravid BPH females, mechanical wounding, and treatment with JA or ABA. Silencing OsJMJ715 enhanced BPH-elicited levels of ABA, JA, and JA-Ile as well as the amount of callose deposition in plants, which in turn increased the resistance of rice to BPH by reducing the feeding of BPH and the hatching rate of BPH eggs. These findings suggest that OsJMJ715 negative regulates the BPH-induced biosynthesis of ABA, JA, and JA-Ile and that BPH benefits by enhancing the expression of OsJMJ715.     

Use of chemical variation and predation as plant defenses byEncelia farinosa against a specialist herbivore

Larvae of the monophagous herbivore,Trirhabda geminata, selectively eat particular plants and plant parts of its natural host,Encelia farinosa. Measurements of leaf damage and larval positions on branches through time support this observation. Time-lapse movie photography revealed that larvae are sufficiently mobile to search most of a plant in a 48-hr period and that aggregations were the result of larval activity and not directly the result of oviposition. Experiments withT. geminata larvae on artificial diets containing a range of natural concentrations of chemical extracts fromE. farinosa leaves showed that the larvae grew significantly slower and had a lower overall survivorship at the high concentration. Combining the results of all choice tests, larvae appeared unable to distinguish between high- and low-concentration agar diets. Considered individually, larval preferences for natural production concentrations changed as the season progressed. Early-season larvae preferred low-concentration leaves, while late-season larvae preferred high-concentrations. Measurements of chemical and nitrogen content of leaves selected by larvae in the field confirmed this pattern. Percent parasitism in field-collected larvae increased with season as the larval population decreased. This combination of slowed growth and increasing parasitism and predation is a putative defense strategy ofEncelia farinosa to prevent adaptation by a specialist herbivore to the total range of compounds elaborated.     

Induced Responses to Herbivory and Jasmonate in Three Milkweed Species

We studied constitutive and induced defensive traits (latex exudation, cardenolides, proteases, and C/N ratio) and resistance to monarch caterpillars (Danaus plexippus) in three closely related milkweed species (Asclepias angustifolia, A. barjoniifolia and A. fascicularis). All traits showed significant induction in at least one of the species. Jasmonate application only partially mimicked the effect of monarch feeding. We found some correspondence between latex and cardenolide content and reduced larval growth. Larvae fed cut leaves of A. angustifolia grew better than larvae fed intact plants. Addition of the cardenolide digitoxin to cut leaves reduced larval growth but ouabain (at the same concentration) had no effect. We, thus, confirm that latex and cardenolides are major defenses in milkweeds, effective against a specialist herbivore. Other traits such as proteases and C/N ratio additionally may be integrated in the defense scheme of those plants. Induction seems to play an important role in plants that have an intermediate level of defense, and we advocate incorporating induction as an additional axis of the plant defense syndrome hypothesis.     

Defensive Role of Tomato Polyphenol Oxidases against Cotton Bollworm (<Emphasis Type="Italic">Helicoverpa armigera</Emphasis>) and Beet Armyworm (<Emphasis Type="Italic">Spodoptera exigua</Emphasis>)

Tomato (Solanum lycopersicum) polyphenol oxidases (PPOs), enzymes that oxidize phenolics to quinones, have been implicated in plant resistance to insects. The role of PPO in resistance to cotton bollworm [Helicoverpa armigera (Hübner)] and beet armyworm [Spodoptera exigua (Hübner)] (Lepidoptera: Noctuidae) was evaluated. Consumption, weight gains, and mortality of larvae feeding on foliage of transgenic tomato lines overexpressing PPO (OP lines) and of larvae feeding on foliage of transgenic tomato lines with suppressed PPO (SP lines) were compared with consumption, weight gains, and mortality of larvae feeding on non-transformed (NT) plants. Increases in foliage consumption and weight gains were observed for cotton bollworms feeding on leaves of SP plants compared to NT and OP plants. PPO activity was negatively correlated with both weight gains and foliar consumption of cotton bollworm, substantiating the defensive role of PPO against this insect. Similarly, beet armyworm consumed less foliage (both young and old leaves) from OP plants than SP plants. Larvae feeding on OP leaves generally exhibited lower weight gains than those feeding on SP leaves. These results indicate that tomato PPO plays a role in resistance to both cotton bollworm and beet armyworm.     

Jasmonic Acid-Induced Changes in <Emphasis Type="BoldItalic">Brassica oleracea</Emphasis> Affect Oviposition Preference of Two Specialist Herbivores

Jasmonic acid (JA) is a key hormone involved in plant defense responses. The effect of JA treatment of cabbage plants on their acceptability for oviposition by two species of cabbage white butterflies, Pieris rapae and P. brassicae, was investigated. Both butterfly species laid fewer eggs on leaves of JA-treated plants compared to control plants. We show that this is due to processes in the plant after JA treatment rather than an effect of JA itself. The oviposition preference for control plants is adaptive, as development time from larval hatch until pupation of P. rapae caterpillars was longer on JA-treated plants. Total glucosinolate content in leaf surface extracts was similar for control and treated plants; however, two of the five glucosinolates were present in lower amounts in leaf surface extracts of JA-treated plants. When the butterflies were offered a choice between the purified glucosinolate fraction isolated from leaf surface extracts of JA-treated plants and that from control plants, they did not discriminate. Changes in leaf surface glucosinolate profile, therefore, do not seem to explain the change in oviposition preference of the butterflies after JA treatment, suggesting that as yet unknown infochemicals are involved.     

Defense Suppression through Interplant Communication Depends on the Attacking Herbivore Species

In response to herbivory, plants emit volatile compounds that play important roles in plant defense. Herbivore-induced plant volatiles (HIPVs) can deter herbivores, recruit natural enemies, and warn other plants of possible herbivore attack. Following HIPV detection, neighboring plants often respond by enhancing their anti-herbivore defenses, but a recent study found that herbivores can manipulate HIPV-interplant communication for their own benefit and suppress defenses in neighboring plants. Herbivores induce species-specific blends of HIPVs and how these different blends affect the specificity of plant defense responses remains unclear. Here we assessed how HIPVs from zucchini plants (Cucurbita pepo) challenged with different herbivore species affect resistance in neighboring plants. Volatile “emitter” plants were damaged by one of three herbivore species: saltmarsh caterpillars (Estigmene acrea), squash bugs (Anasa tristis), or striped cucumber beetles (Acalymma vittatum), or were left as undamaged controls. Neighboring “receiver” plants were exposed to HIPVs or control volatiles and then challenged by the associated herbivore species. As measures of plant resistance, we quantified herbivore feeding damage and defense-related phytohormones in receivers. We found that the three herbivore species induced different HIPV blends from squash plants. HIPVs induced by saltmarsh caterpillars suppressed defenses in receivers, leading to greater herbivory and lower defense induction compared to controls. In contrast, HIPVs induced by cucumber beetles and squash bugs did not affect plant resistance to subsequent herbivory in receivers. Our study shows that herbivore species identity affects volatile-mediated interplant communication in zucchini, revealing a new example of herbivore defense suppression through volatile cues.

     

Herbivore-Induced Defenses in Tomato Plants Enhance the Lethality of the Entomopathogenic Bacterium, <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> var. <Emphasis Type="Italic">kurstaki</Emphasis>

Plants can influence the effectiveness of microbial insecticides through numerous mechanisms. One of these mechanisms is the oxidation of plant phenolics by plant enzymes, such as polyphenol oxidases (PPO) and peroxidases (POD). These reactions generate a variety of products and intermediates that play important roles in resistance against herbivores. Oxidation of the catecholic phenolic compound chlorogenic acid by PPO enhances the lethality of the insect-killing bacterial pathogen, Bacillus thuringiensis var. kurstaki (Bt) to the polyphagous caterpillar, Helicoverpa zea. Since herbivore feeding damage often triggers the induction of higher activities of oxidative enzymes in plant tissues, here we hypothesized that the induction of plant defenses would enhance the lethality of Bt on those plants. We found that the lethality of a commercial formulation of Bt (Dipel® PRO DF) on tomato plants was higher if it was applied to plants that were induced by H. zea feeding or induced by the phytohormone jasmonic acid. Higher proportions of H. zea larvae killed by Bt were strongly correlated with higher levels of PPO activity in the leaflet tissue. Higher POD activity was only weakly associated with higher levels of Bt-induced mortality. While plant-mediated variation in entomopathogen lethality is well known, our findings demonstrate that plants can induce defensive responses that work in concert with a microbial insecticide/entomopathogen to protect against insect herbivores.     

Fungal Infection Reduces Herbivore-Induced Plant Volatiles of Maize but does not Affect Naïve Parasitoids

Plants attacked by insects release volatile compounds that attract the herbivores' natural enemies. This so-called indirect defense is plastic and may be affected by an array of biotic and abiotic factors. We investigated the effect of fungal infection as a biotic stress agent on the emission of herbivore-induced volatiles and the possible consequences for the attraction of two parasitoid species. Maize seedlings that were simultaneously attacked by the fungus Setosphaeria turcica and larvae of Spodoptera littoralis emitted a blend of volatiles that was qualitatively similar to the blend emitted by maize that was damaged by only the herbivore, but there was a clear quantitative difference. When simultaneously challenged by fungus and herbivore, the maize plants emitted in total 47% less of the volatiles. Emissions of green leaf volatiles were unaffected. In a six-arm olfactometer, the parasitoids Cotesia marginiventris and Microplitis rufiventris responded equally well to odors of herbivore-damaged and fungus- and herbivore-damaged maize plants. Healthy and fungus-infected plants were not attractive. An additional experiment showed that the performance of S. littoralis caterpillars was not affected by the presence of the pathogen, nor was there an effect on larvae of M. rufiventris developing inside the caterpillars. Our results confirm previous indications that naïve wasps may respond primarily to the green leaf volatiles.     

Jasmonate in Lepidopteran Larvae

Jasmonic acid (JA) is a key molecule initiating plant defensive responses to herbivory. Our previous work has shown that this phytohormone is often present at high concentrations in eggs and neonates of lepidopteran species. In this work, we document the concentrations of JA found in various tissues of larval H. virescens fed on artificial diets with varying JA concentrations and on transgenic tobacco lines with different capabilities of producing JA. At high JA concentrations, excess jasmonate appeared to be voided in frass, but significant amounts were found in most larval tissues we analyzed as well as in regurgitant. At lower concentrations, caterpillars excreted less JA and appeared to accumulate relatively greater amounts of JA in their tissues. In both experiments, substantial amounts of JA were found in labial salivary glands, a notable result since JA is an important signaling molecule involved in the induction of plant defenses.     

Brassica Plant Responses to Mild Herbivore Stress Elicited by Two Specialist Insects from Different Feeding Guilds

Compensation growth and chemical defense are two components of plant defense strategy against herbivores. In this study, compensation growth and the response of primary and secondary metabolites were investigated in Brassica rapa plants subjected to infestation by two herbivores from contrasting feeding guilds, the phloem-feeding aphid Brevicoryne brassicae and the leaf-feeding caterpillar Pieris brassicae. These specialist herbivores were used at two different densities and allowed to feed for seven days on a young caged leaf. Changes in growth rates were assessed for total leaf area and bulb mass, whereas changes in primary and secondary metabolites were evaluated in young and mature leaves, roots, and bulbs. Mild stress by caterpillars on young plants enhanced mean bulb mass and elicited a contrasting regulation of aliphatic and indolic glucosinolates in the leaves. In contrast, mild stress by aphids enhanced leaf growth and increased glucosinolate concentrations in the bulb, the most important storage organ of B. rapa. A similar mild stress by either herbivore to older plants did not alter plant growth parameters or concentrations of the metabolites analyzed. In conclusion, Brassica plant growth was either maintained or enhanced under mild herbivore stress, and defense patterns differed strongly in response to herbivore type and plant development stage. These results have implications for the understanding of plasticity in plant defenses against herbivores and for the management of Brassica rapa in agroecosystems.     

Host Plant Suitability in a Specialist Herbivore, <Emphasis Type="Italic">Euphydryas anicia</Emphasis> (Nymphalidae): Preference,Performance and Sequestration

The checkerspot butterfly, Euphydryas anicia (Nymphalidae), specializes on plants containing iridoid glycosides and has the ability to sequester these compounds from its host plants. This study investigated larval preference, performance, and sequestration of iridoid glycosides in a population of E. anicia at Crescent Meadows, Colorado, USA. Although previous studies showed that other populations in Colorado use the host plant, Castilleja integra (Orobanchaceae), we found no evidence for E. anicia ovipositing or feeding on C. integra at Crescent Meadows. Though C. integra and another host plant, Penstemon glaber (Plantaginaceae), occur at Crescent Meadows, the primary host plant used was P. glaber. To determine why C. integra was not being used at the Crescent Meadows site, we first examined the host plant preference of naïve larvae between P. glaber and C. integra. Then we assessed the growth and survivorship of larvae reared on each plant species. Finally, we quantified the iridoid glycoside concentrations of the two plant species and diapausing caterpillars reared on each host plant. Our results showed that E. anicia larvae prefer P. glaber. Also, larvae survive and grow better when reared on P. glaber than on C. integra. Castilleja integra was found to contain two primary iridoid glycosides, macfadienoside and catalpol, and larvae reared on this plant sequestered both compounds; whereas P. glaber contained only catalpol and larvae reared on this species sequestered catalpol. Thus, although larvae are able to use C. integra in the laboratory, the drivers behind the lack of use at the Crescent Meadows site remain unclear.     

Age-Dependent Effects of Jasmonic Acid Treatment and Wind Exposure on Foliar Oxidase Activity and Insect Resistance in Tomato

Jasmonic acid (JA) treatment of tomato plants induces several defense-related oxidative enzymes and increases pest resistance in a manner thought to simulate natural insect wounding. In a full-factorial greenhouse experiment, we examined the independent and interactive effects of plant age and exposure to wind-induced mechanical stress (MS), on the ability of JA to induce defense in tomato. In general, treatment of 4-, 6-, and 8-week-old tomato plants with 1 mM JA resulted in the induction of peroxidase and polyphenol oxidase activity and reduced the relative growth rate of first-instar Manduca sexta larvae fed treated leaves, in accordance with other studies. Peroxidase activity increased with plant age and was induced by JA most strongly in older plants. In contrast, polyphenol oxidase activity did not change with plant age and was induced by JA most strongly in young plants. While relative growth rates of M. sexta were lower on older plants overall, JA reduced growth rates most strongly in young plants, in which JA treatment enhanced polyphenol oxidase activity by more than 70%. MS enhanced the activity of peroxidase, but substantially reduced the activity of polyphenol oxidase; the latter most intensely on older plants. M. sexta tended to grow more slowly on MS-treated plants, although this effect was not significant. Thus, reduced polyphenol oxidase activity in MS-treated plants did not lead to an increase in growth rate of M. sexta, possibly because peroxidase activity was still elevated in MS-treated plants. Significant interactions between JA and MS and three-way interactions were not detected for any variable, although the inductive effects of both JA and MS interacted in complex ways with plant age. Our results indicate that resistance traits in tomato are differentially affected by JA and wind exposure and differ in their relative contribution to defense as plants age.