Caterpillar Oral Secretions Research Articles

Foliar herbivory‐enhanced mycorrhization is associated with increased levels of lipids in root and root exudates

Abstract Insect herbivory can affect interactions between plants and arbuscular mycorrhizal (AM) fungi through herbivore‐modified root carbon pools, while the specific metabolic changes underlying fungal responses to herbivory are poorly understood. Here we explored the impacts of foliar herbivory and mechanical wounding on AM colonisation and AM community composition of common ragweed (Ambrosia artemisiifolia) and the role of root metabolites in mediating these effects. Foliar insect herbivory enhanced AM colonisation, whereas mechanical wounding only enhanced AM colonisation in combination with application of caterpillar oral secretions. Meanwhile, the relative abundance of Glomus species was increased in root endosphere, rhizoplane and rhizosphere soils after foliar herbivory. Foliar herbivory also increased the concentrations of fatty acids in roots but decreased phenolics, and their concentrations were significantly correlated with AM colonisation. Addition of exudates from plants exposed to herbivory resulted in increases in AM colonisation of plants without herbivory. Moreover, widely targeted metabolomic analyses revealed that foliar herbivory enhanced the relative abundance of lipids and decreased phenols in root exudates. Synthesis. We show that plants can enhance their associations with arbuscular mycorrhizal (AM) fungi when subject to above‐ground herbivory, possibly mediated by herbivore‐induced increases in the levels of root lipids. Our findings highlight the role of root lipids in above‐below‐ground biological interactions, providing novel insights into plant‐AM fungi integrative responses to biotic stresses.

Metabolomic fingerprint of cabbage resistance to Mamestra brassicae L. (Lepidoptera: Noctuidae).

Plants defend themselves from insect feeding by activating specific metabolic pathways. We performed a metabolomic analysis to compare the metabolome reorganization that occurs in the leaves of two genotypes of cabbage (one partially resistant and one susceptible) when attacked by Mamestra brassicae caterpillars. The comparison of the metabolomic reorganization of both genotypes allowed us to identify 43 metabolites that are specifically associated with the insect feeding response in the resistant genotype. Of these, 19% are lipids or lipid-related compounds, most of which are modified fatty acids. These include glycosylated, glycerol-binding and oxidized fatty acids, the majority being associated with the oxylipin pathway. Some of the identified lipids are unlikely to be produced by plants and may be the result of biochemical reactions in the caterpillar oral secretions. A further 16% are phenylpropanoids. Interestingly, some phenylpropanoids were not present in the susceptible genotype, making them possible candidates for specific resistance-related compounds. Our results suggest that glucosinolates do not have a clear role in the resistance to M. brassicae feeding on cabbage. Using an untargeted metabolomics approach, we associated the regulation of metabolic pathways related to lipid signalling and phenylpropanoid compounds with the resistance to this pest. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A highly accumulated secretory protein from cotton bollworm interacts with basic helix-loop-helix transcription factors to dampen plant defense.

Caterpillar oral secretion (OS) contains active molecules that modulate plant defense signaling. We isolated an effector-like protein (Highly Accumulated Secretory Protein 1, HAS1) from cotton bollworm (Helicoverpa armigera) that is the most highly accumulated secretory protein of the nondigestive components in OS and belongs to venom R-like protein. Elimination of HAS1 by plant-mediated RNA interference reduced the suppression of OS on the defense response in plants. Plants expressing HAS1 are more susceptible to insect herbivory accompanied by the reduced expressions of multiple defense genes. HAS1 binds to the basic helix-loop-helix (bHLH) transcription factors, including GoPGF involved in pigmented gland formation and defense compounds biosynthesis in cotton and MYC3/MYC4 the main regulators in jasmonate (JA) signaling in Arabidopsis. The binding activity is required for HAS1 to inhibit the activation of bHLHs on plant defense gene expressions. Together with our previous study that another venom R-like protein HARP1 in cotton bollworm OS blocks JA signaling by interacting with JASMONATE-ZIM-domain repressors, we conclude that the venom R-like proteins in OS interfere with plant defense in a dual suppression manner. Considering the venom proteins in parasitic wasp assault the immune system of its host animal, our investigation reveals their conserved function in carnivorous and herbivorous insects.

Plant-phenotypic changes induced by parasitoid ichnoviruses enhance the performance of both unparasitized and parasitized caterpillars.

There is increasing awareness that interactions between plants and insects can be mediated by microbial symbionts. Nonetheless, evidence showing that symbionts associated with organisms beyond the second trophic level affect plant‐insect interactions are restricted to a few cases belonging to parasitoid‐associated bracoviruses. Insect parasitoids harbour a wide array of symbionts which, like bracoviruses, can be injected into their herbivorous hosts to manipulate their physiology and behaviour. Yet, the function of these symbionts in plant‐based trophic webs remains largely overlooked. Here, we provide the first evidence of a parasitoid‐associated symbiont belonging to the group of ichnoviruses which affects the strength of plant‐insect interactions. A comparative proteomic analysis shows that, upon parasitoid injection of calyx fluid containing ichnovirus particles, the composition of salivary glands of caterpillars changes both qualitatively (presence of two viral‐encoded proteins) and quantitatively (abundance of several caterpillar‐resident enzymes, including elicitors such as glucose oxidase). In turn, plant phenotypic changes triggered by the altered composition of caterpillar oral secretions affect the performance of herbivores. Ichnovirus manipulation of plant responses to herbivory leads to benefits for their parasitoid partners in terms of reduced developmental time within the parasitized caterpillar. Interestingly, plant‐mediated ichnovirus‐induced effects also enhance the performances of unparasitized herbivores which in natural conditions may feed alongside parasitized ones. We discuss these findings in the context of ecological costs imposed to the plant by the viral symbiont of the parasitoid. Our results provide intriguing novel findings about the role played by carnivore‐associated symbionts on plant‐insect‐parasitoid systems and underline the importance of placing mutualistic associations in an ecological perspective.

A receptor-like protein mediates plant immune responses to herbivore-associated molecular patterns.

Herbivory is fundamental to the regulation of both global food webs and the extent of agricultural crop losses. Induced plant responses to herbivores promote resistance and often involve the perception of specific herbivore-associated molecular patterns (HAMPs); however, precisely defined receptors and elicitors associated with herbivore recognition remain elusive. Here, we show that a receptor confers signaling and defense outputs in response to a defined HAMP common in caterpillar oral secretions (OS). Staple food crops, including cowpea (Vigna unguiculata) and common bean (Phaseolus vulgaris), specifically respond to OS via recognition of proteolytic fragments of chloroplastic ATP synthase, termed inceptins. Using forward-genetic mapping of inceptin-induced plant responses, we identified a corresponding leucine-rich repeat receptor, termed INR, specific to select legume species and sufficient to confer inceptin-induced responses and enhanced defense against armyworms (Spodoptera exigua) in tobacco. Our results support the role of plant immune receptors in the perception of chewing herbivores and defense.

Tobacco Hornworm (Manduca sexta) Oral Secretion Elicits Reactive Oxygen Species in Isolated Tomato Protoplasts.

Plants are under constant attack by a suite of insect herbivores. Over millions of years of coexistence, plants have evolved the ability to sense insect feeding via herbivore-associated elicitors in oral secretions, which can mobilize defense responses. However, herbivore-associated elicitors and the intrinsic downstream modulator of such interactions remain less understood. In this study, we show that tobacco hornworm caterpillar (Manduca sexta) oral secretion (OS) induces reactive oxygen species (ROS) in tomato (Solanum lycopersicum) protoplasts. By using a dye-based ROS imaging approach, our study shows that application of plant-fed (PF) M. sexta OS generates significantly higher ROS while artificial diet-fed (DF) caterpillar OS failed to induce ROS in isolated tomato protoplasts. Elevation in ROS generation was saturated after ~140 s of PF OS application. ROS production was also suppressed in the presence of an antioxidant NAC (N-acetyl-L-cysteine). Interestingly, PF OS-induced ROS increase was abolished in the presence of a Ca2+ chelator, BAPTA-AM (1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid). These results indicate a potential signaling cascade involving herbivore-associated elicitors, Ca2+, and ROS in plants during insect feeding. In summary, our results demonstrate that plants incorporate a variety of independent signals connected with their herbivores to regulate and mount their defense responses.

Distinct Arabidopsis Responses to Two Generalist Caterpillar Species Differing in Host Breadth

In general, caterpillar herbivores with a narrow host preference (specialists) have evolved mechanisms to circumvent specific plant defenses. In contrast, caterpillars with a broader host range (generalists) may manipulate phytohormone pathways common to many plant species to attenuate induced defenses. Many studies have compared plant responses to specialist versus generalist caterpillars. In contrast, this study evaluates the induced response of Arabidopsis thaliana to two generalist caterpillar species, the cabbage looper, Trichoplusia ni, and the beet armyworm, Spodoptera exigua. Although both caterpillars are considered generalists, S. exigua has a broader plant host range, whereas T. ni prefers Brassicaceous plants. Our study shows that most responses to caterpillar herbivory, such as the jasmonate burst, are similar in plants attacked by either insect species; however, we do observe dynamic and temporal differences in specific responses. Expression of AtZAT10, a 12-oxo-phytodienoic acid-responsive gene, is only induced in response to T. ni damage. In comparison, only S. exigua herbivory activates the salicylic acid/NPR1-dependent pathway, as observed by the expression of the marker gene AtPR1. Even though both species induce AtPDF1.2 expression, we found caterpillar-specific temporal differences: T. ni herbivory results in sustained expression over time, whereas gene expression is sharply downregulated at 36 h in S. exigua-attacked plants. Although damage by these two caterpillar species induced AtMYB28 and AtMYB34 expression, specific short- and long-chain aliphatic and indolic glucosinolates accumulate only in response to S. exigua herbivory. These species-specific, plant-induced responses likely reflect differences in effectors found in caterpillar oral secretions. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Effects of soil salinity on the expression of direct and indirect defences in wild cotton Gossypium hirsutum

Abstract Previous studies have frequently reported effects of abiotic factors on herbivore‐induced plant defences based on effects on single plant traits. However, plants commonly express multiple defences simultaneously and these traits are often correlated. Thus, a fuller understanding of abiotic‐context dependency in plant defence requires measuring multiple traits and addressing their patterns of correlated expression. We evaluated the effects of soil salinity on the expression of direct (phenolic compounds, gossypol gland density) and indirect (volatile organic compounds, extrafloral nectar) defensive traits in wild cotton Gossypium hirsutum. Specifically, we asked whether soil salinity affects the induction of these traits, and whether it shapes trait correlations potentially underlying altered patterns of trait induction. We conducted a factorial experiment with 16 cotton genotypes where we manipulated soil salinity and defence induction by applying artificial leaf damage (25% mechanical damage and caterpillar oral secretions) and measured defence levels at different time points post damage. Leaf damage induced most traits except gossypol gland density, whereas salinity did not have a mean effect (across constitutive and induced levels) on any of the measured traits. Nonetheless, salinity prevented the induction of phenolic compounds (condensed and hydrolysable tannins), and also affected trait correlations. Specifically, phenolic compounds were negatively associated with nectar production only under salinized conditions, an apparent trade‐off that could affect the induction of phenolic compounds. In addition, positive correlations between phenolic compounds and gland density and root biomass observed under control conditions were lost under salinized conditions. Synthesis. By investigating the effects of soil salinity on the expression of multiple direct and indirect defensive traits and their underlying correlations, these findings build towards a better understanding of how abiotic context dependency shapes plant allocation to and expression of multiple defensive traits.

Caterpillar Oral Secretion Elicits Reactive Oxygen Species in Isolated Plant Protoplasts

Over millions of years, plants have evolved and developed several defense strategies such as spines, trichomes, toxins, and plant volatiles to either directly or indirectly defend against insect herbivores. Reactive Oxygen Species (ROS) is a signaling molecule that is suspected of signaling herbivory by activating defense cascade at the cellular level, leading to local and systemic defense gene expression. In this study, we show that tobacco hornworm caterpillar (Manduca sexta) oral secretion (OS) induces ROS in tomato (Solanum lycopersicum) protoplasts. We used a dye-based ROS imaging approach to dissect the interaction between a damaging herbivore and its host plant. Our study shows that application of Plant-Fed (PF) caterpillar OS increased ROS generation while artificial Diet-Fed (DF) caterpillar OS failed to induce ROS in isolated tomato protoplasts. The elevation in ROS generation was observed after ˜100 s and was saturated after 165 s of PF-OS application. Interestingly, the PF-OS induced ROS increase was abolished in the presence of a Ca2+ chelator BAPTA-AM, suggesting possible crosstalk between Ca2+ and ROS signaling. Targeting the pathway that triggers ROS generation could be used to develop a herbivore origin potent elicitor of plant defense gene expression (ROS elicitor) pest management research.

Compensation to simulated insect leaf herbivory in wild cotton (Gossypium hirsutum): responses to multiple levels of damage and associated traits.

Identifying the mechanisms of compensation to insect herbivory remains a major challenge in plant biology and evolutionary ecology. Most previous studies have addressed plant compensatory responses to one or two levels of insect herbivory, and the underlying traits mediating such responses remain elusive in many cases. We evaluated responses associated with compensation to multiple intensities of leaf damage (0% control, 10%, 25%, 50%, 75% of leaf area removed) by means of mechanical removal of foliar tissue and application of a caterpillar (Spodoptera exigua) oral secretions in 3-month-old wild cotton plants (Gossypium hirsutum). Four weeks post-treatment, we measured plant growth and multiple traits associated with compensation, namely: changes in above- and belowground, biomass and the concentration of nutrients (nitrogen and phosphorus) and non-structural carbon reserves (starch and soluble sugars) in roots, stems and leaves. We found that wild cotton fully compensated in terms of growth and biomass allocation when leaf damage was low (10%), whereas moderate (25%) to high leaf damage in some cases led to under-compensation. Nonetheless, high levels of leaf removal (50% and 75%) in most cases did not cause further reductions in height and allocation to leaf and stem biomass relative to low and moderate damage. There were significant positive effects of leaf damage on P concentration in leaves and stems, but not roots, as well as a negative effect on soluble sugars in roots. These results indicate that wild cotton fully compensated for a low level of leaf damage but under-compensated under moderate to high leaf damage, but can nonetheless sustain growth despite increasing losses to herbivory. Such responses were possibly mediated by a re-allocation of carbohydrate reserves from roots to shoots.

Optimizing Crops for Biocontrol of Pests and Disease.

Volatile compounds and extrafloral nectar are common defenses of wild plants; however, in crops they bear an as-yet underused potential for biological control of pests and diseases. Odor emission and nectar secretion are multigene traits in wild plants, and thus form difficult targets for breeding. Furthermore, domestication has changed the capacity of crops to express these traits. We propose that breeding crops for an enhanced capacity for tritrophic interactions and volatile-mediated direct resistance to herbivores and pathogens can contribute to environmentally-friendly and sustainable agriculture. Natural plant volatiles with antifungal or repellent properties can serve as direct resistance agents. In addition, volatiles mediating tritrophic interactions can be combined with nectar-based food rewards for carnivores to boost indirect plant defense.

Effect of sequential induction by Mamestra brassicae L. and Tetranychus urticae Koch on lima bean plant indirect defense.

Attack by multiple herbivores often leads to modification of induced plant defenses compared to single herbivory, yet little is known about the effects on induced indirect plant defense. Here, we investigated the effect of sequential induction of plant defense by Mamestra brassicae caterpillar oral secretion and an infestation by Tetranychus urticae spider mites on the expression of indirect plant defense in Lima bean plants. The effect on indirect defense was assessed using behavior assays with the specialist predatory mite Phytoseiulus persimilis in an olfactometer, headspace analysis of 11 major herbivore-induced plant volatiles (HIPVs) including (E)-β-ocimene, and transcript levels of the corresponding gene Phaseolus lunatus (E)-β-ocimene synthase (PlOS). Predatory mites were found to distinguish between plants induced by spider mites and caterpillar oral secretion but not between plants with single spider mite infestation and plants induced by caterpillar oral secretion prior to spider mite infestation. Indeed, the volatile blends emitted by plants induced by spider mites only and the sequential induction treatment of caterpillar oral secretion followed by spider mite infestation, were similar. Our results suggest that plant indirect defense is not affected by previous treatment with oral secretion of M. brassicae caterpillars.

Herbivore Cues from the Fall Armyworm (Spodoptera frugiperda) Larvae Trigger Direct Defenses in Maize

In addition to feeding damage, herbivores release cues that are recognized by plants to elicit defenses. Caterpillar oral secretions have been shown to trigger herbivore defense responses in several different plant species. In this study, the effects of two fall armyworm (Spodoptera frugiperda) oral secretions (saliva and regurgitant) on caterpillar defense responses in maize (Zea mays) were examined. Only minute amounts of regurgitant were deposited on the maize leaf during larval feeding bouts and its application to leaves failed to induce the expression of several herbivore defense genes. On the other hand, caterpillars consistently deposited saliva on leaves during feeding and the expression of several maize defense genes significantly increased in response to saliva application and larval feeding. However, feeding by ablated caterpillars with impaired salivation did not induce these defenses. Furthermore, bioassays indicated that feeding by unablated caterpillars significantly enhanced defenses when compared with that of ablated caterpillars. Another critical finding was that the maize genotype and stage of development affected the expression of defense genes in response to wounding and regurgitant treatments. These results demonstrate that fall armyworm saliva contains elicitors that trigger herbivore defenses in maize.

Caterpillar- and Salivary-Specific Modification of Plant Proteins

Though there is overlap, plant responses to caterpillar herbivory show distinct variations from mechanical wounding. In particular, effectors in caterpillar oral secretions modify wound-associated plant responses. Previous studies have focused on transcriptional and protein abundance differences in response to caterpillar herbivory. This study investigated Spodoptera exigua caterpillar-specific post-translational modification of Arabidopsis thaliana soluble leaf proteins by liquid chromatography/electrospray ionization/mass spectroscopy/mass spectroscopy (LC/ESI/MS/MS). Given that caterpillar labial saliva contains oxidoreductases, such as glucose oxidase, particular attention was paid to redox-associated modifications, such as the oxidation of protein cysteine residues. Caterpillar- and saliva-specific protein modifications were observed. Differential phosphorylation of the jasmonic acid biosynthetic enzyme, lipoxygenase 2, and a chaperonin protein is seen in plants fed upon by caterpillars with intact salivary secretions compared to herbivory by larvae with impaired labial salivary secretions. Often a systemic suppression of photosynthesis is associated with caterpillar herbivory. Of the five proteins modified in a caterpillar-specific manner (a transcription repressor, a DNA-repair enzyme, PS I P700, Rubisco and Rubisco activase), three are associated with photosynthesis. Oxidative modifications are observed, such as caterpillar-specific denitrosylation of Rubisco activase and chaperonin, cysteine oxidation of Rubisco, DNA-repair enzyme, and chaperonin and caterpillar-specific 4-oxo-2-nonenal modification of the DNA-repair enzyme.

Transcriptome profiles of hybrid poplar (Populus trichocarpa × deltoides) reveal rapid changes in undamaged, systemic sink leaves after simulated feeding by forest tent caterpillar (Malacosoma disstria)

• Poplar has been established as a model tree system for genomic research of the response to biotic stresses. This study describes a series of induced transcriptome changes and the associated physiological characterization of local and systemic responses in hybrid poplar (Populus trichocarpa × deltoides) after simulated herbivory. • Responses were measured in local source (LSo), systemic source (SSo), and systemic sink (SSi) leaves following application of forest tent caterpillar (Malacosoma disstria) oral secretions to mechanically wounded leaves. • Transcriptome analyses identified spatially and temporally dynamic, distinct patterns of local and systemic gene expression in LSo, SSo and SSi leaves. Galactinol synthase was strongly and rapidly upregulated in SSi leaves. Genome analyses and full-length cDNA cloning established an inventory of poplar galactinol synthases. Induced changes of galactinol and raffinose oligosaccharides were detected by anion-exchange high-pressure liquid chromatography. • The LSo leaves showed a rapid and strong transcriptome response compared with a weaker and slower response in adjacent SSo leaves. Surprisingly, the transcriptome response in distant, juvenile SSi leaves was faster and stronger than that observed in SSo leaves. Systemic transcriptome changes of SSi leaves have signatures of rapid change of metabolism and signaling, followed by later induction of defense genes.

Fatty Acid-amino Acid Conjugates Diversification in Lepidopteran Caterpillars

Fatty acid amino acid conjugates (FACs) have been found in noctuid as well as sphingid caterpillar oral secretions; in particular, volicitin [N-(17-hydroxylinolenoyl)-L-glutamine] and its biochemical precursor, N-linolenoyl-L-glutamine, are known elicitors of induced volatile emissions in corn plants. These induced volatiles, in turn, attract natural enemies of the caterpillars. In a previous study, we showed that N-linolenoyl-L-glutamine in larval Spodoptera litura plays an important role in nitrogen assimilation which might be an explanation for caterpillars synthesizing FACs despite an increased risk of attracting natural enemies. However, the presence of FACs in lepidopteran species outside these families of agricultural interest is not well known. We conducted FAC screening of 29 lepidopteran species, and found them in 19 of these species. Thus, FACs are commonly synthesized through a broad range of lepidopteran caterpillars. Since all FAC-containing species had N-linolenoyl-L-glutamine and/or N-linoleoyl-L-glutamine in common, and the evolutionarily earliest species among them had only these two FACs, these glutamine conjugates might be the evolutionarily older FACs. Furthermore, some species had glutamic acid conjugates, and some had hydroxylated FACs. Comparing the diversity of FACs with lepidopteran phylogeny indicates that glutamic acid conjugates can be synthesized by relatively primitive species, while hydroxylation of fatty acids is limited mostly to larger and more developed macrolepidopteran species.

Systemic induction of volatile release in cotton: How specific is the signal to herbivory?

Plants attacked by herbivorous insects release chemical signals that attract natural enemies of the herbivores to the damaged plants. Feeding of Spodoptera exigua larvae on the lower leaves of cotton (Gossypium hirsutum L.) for multiple feeding periods of 9-12 h with a 12 h, interval in between when the caterpillars are removed overnight, will induce a systemic release of volatile compounds that is comparable to the volatiles released in response to continuous feeding damage on the lower leaves for several days. The systemic volatile release in response to herbivory can be mimicked by mechanically damaging the lower leaves and applying caterpillar oral secretion to the injured leaves over 4 days. Cotton plants that are only mechanically damaged systemically release significantly less beta-pinene, myrcene, (Z)-3-hexenyl acetate, (E)-beta-farnesene and (E,E)-alpha-farnesene after 4 days compared to plants damaged mechanically with application of caterpillar regurgitant. However, multiple 9-12 h mechanical damage alone induces a significantly higher systemic release of (Z)-3-hexenyl acetate, myrcene, (E)-beta-ocimene, and (E)-beta-farnesene after 4 days compared to undamaged control plants. This indicates that multiple mechanical damage alone cannot mimic completely the response induced by mechanically injuring the leaves and applying caterpillar regurgitant. A specific elicitor in the regurgitant of the caterpillar enhances the amount of several systemically released volatiles. Thus, the systemic release of volatile compounds by herbivore-damaged cotton plants appears to be regulated by at least two different mechanisms.

Nitrogen deficiency increases volicitin-induced volatile emission, jasmonic acid accumulation, and ethylene sensitivity in maize.

Insect herbivore-induced plant volatile emission and the subsequent attraction of natural enemies is facilitated by fatty acid-amino acid conjugate (FAC) elicitors, such as volicitin [N-(17-hydroxylinolenoyl)-L-glutamine], present in caterpillar oral secretions. Insect-induced jasmonic acid (JA) and ethylene (E) are believed to mediate the magnitude of this variable response. In maize (Zea mays) seedlings, we examined the interaction of volicitin, JA, and E on the induction of volatile emission at different levels of nitrogen (N) availability that are known to influence E sensitivity. N availability and volicitin-induced sesquiterpene emission are inversely related as maximal responses were elicited in N-deficient plants. Plants with low N availability demonstrated similar volatile responses to volicitin (1 nmol plant(-1)) and JA (100 nmol plant(-1)). In contrast, plants with medium N availability released much lower amounts of volicitin-induced sesquiterpenes compared with JA, suggesting an alteration in volicitin-induced JA levels. As predicted, low N plants exhibited greater sustained increases in wound- and volicitin-induced JA levels compared with medium N plants. N availability also altered volicitin-E interactions. In low N plants, E synergized volicitin-induced sesquiterpene and indole emission 4- to 12-fold, with significant interactions first detected at 10 nL L(-1) E. Medium N plants demonstrated greatly reduced volicitin-E interactions. Volicitin-induced sesquiterpene emission was increased by E and was decreased by pretreatment the E perception inhibitor 1-methylcyclopropene without alteration in volicitin-induced JA levels. N availability influences plant responses to insect-derived elicitors through changes in E sensitivity and E-independent JA kinetics.

Synergistic interactions between volicitin, jasmonic acid and ethylene mediate insect-induced volatile emission in Zea mays.

Plants display differential responses following mechanical damage and insect herbivory. Both caterpillar attack and the application of caterpillar oral secretions (OS) to wounded leaves stimulates volatile emission above mechanical damage alone. Volicitin (N-17-hydroxylinolenoyl-l-glutamine), present in beet armyworm (BAW, Spodoptera exigua) OS, is a powerful elicitor of volatiles in excised maize seedlings (Zea mays cv. Delprim). We consider some of the mechanistic differences between wounding and insect herbivory in maize by examining the activity of volicitin, changes in jasmonic acid (JA) levels, and volatile emission from both intact plant and excised leaf bioassays. Compared to mechanical damage alone, volicitin stimulated increases in both JA levels and sesquiterpene volatiles when applied to intact plants. In a bioassay comparison, excised leaves were more sensitive and produced far greater volatile responses than intact plants following applications of both volicitin and JA. In the excised leaf bioassay, volicitin applications (10-500 pmol) to wounded leaves resulted in dose dependent JA increases and a direct positive relationship between JA and sesquiterpene volatile emission. Interestingly, volicitin-induced JA levels did not differ between intact and excised bioassays, suggesting a possible interaction of JA with other regulatory signals in excised plants. In addition to JA, insect herbivory is known to stimulate the production of ethylene. Significant increases in ethylene were induced only by BAW herbivory and not by either wounding or volicitin treatments. Using intact plant bioassays, ethylene (at 1 micro l l-1 or less) greatly promoted volatile emission induced by volicitin and JA but not mechanical damage alone. For intact plants, wounding, elicitor-induced JA and insect-induced ethylene appear to be important interacting components in the stimulation of insect-induced volatile emission.

De Novo Biosynthesis of Volatiles Induced by Insect Herbivory in Cotton Plants.

In response to insect feeding on the leaves, cotton (Gossypium hirsutum L.) plants release elevated levels of volatiles, which can serve as a chemical signal that attracts natural enemies of the herbivore to the damaged plant. Pulse-labeling experiments with [13C]CO2 demonstrated that many of the volatiles released, including the acyclic terpenes (E,E)-[alpha]-farnesene, (E)-[beta]-farnesene, (E)-[beta]-ocimene, linalool, (E)-4,8-dimethyl-1,3,7-nonatriene, and (E/E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene, as well as the shikimate pathway product indole, are biosynthesized de novo following insect damage. However, other volatile constituents, including several cyclic terpenes, butyrates, and green leaf volatiles of the lipoxygenase pathway are released from storage or synthesized from stored intermediates. Analysis of volatiles from artificially damaged plants, with and without beet armyworm (Spodoptera exigua Hubner) oral secretions exogenously applied to the leaves, as well as volatiles from beet armyworm-damaged and -undamaged control plants, demonstrated that the application of caterpillar oral secretions increased both the production and release of several volatiles that are synthesized de novo in response to insect feeding. These results establish that the plant plays an active and dynamic role in mediating the interaction between herbivores and natural enemies of herbivores.