Nucleopolyhedrovirus infection enhances plant defences by increasing plant volatile diversity

Plant–herbivore–entomopathogen tri-trophic interactions and biodiversity are relatively understudied topics in ecology. Particularly, the effects of entomopathogens on herbivore-induced plant volatiles and plant volatile diversity on the defensive function of plants have not been studied in detail. We used soybean (Glycine max), beet armyworm larvae (Spodoptera exigua), and nucleopolyhedrovirus (NPV) as a tri-trophic system to determine whether NPV infection can promote the emission and diversity of volatiles from plants. We also investigated whether NPV infection affects the attraction of Microplitis pallidipes, an important endoparasitoid of larval S. exigua. Uninfested soybean plants released 7 detectable volatile compounds while plants fed upon by healthy and NPV-infected S. exigua larvae released 12 and 15 volatiles, respectively. Female parasitoids were more attracted to the volatiles from plants that were fed upon by NPV-infected larvae than healthy larvae, and more attracted to the volatiles from plants that were fed upon by healthy larvae than no larvae. The selective responses of parasitoids to plant odours increased as plant volatile diversity increased. Our study suggests that the NPV infection facilitates the release of plant volatiles and enhances the defensive function of plants by increasing plant volatile diversity which in turn attracts more parasitoids. Also, this work reveals that plants might accrue two indirect benefits from NPV infection, cessation of herbivore feeding and more parasitisation.     

Potential role for gut microbiota in cell wall digestion and glucoside detoxification in Tenebrio molitor larvae

Tenebrio molitor larvae were successfully reared free of cultivatable gut lumen bacteria, yeasts and fungi using two approaches; aseptic rearing from surface sterilized eggs and by feeding larvae with antibiotic-containing food. Insects were reared on a rich-nutrient complete diet or a nutrient-poor refractory diet. A comparison of digestive enzyme activities in germ free and conventional insects containing a gut microbiota did not reveal gross differences in enzymes that degrade cell walls from bacteria (lysozyme), fungi (chitinase and laminarinase) and plants (cellulase and licheninase). This suggested that microbial-derived enzymes are not an essential component of the digestive process in this insect. However, more detailed analysis of T. molitor midgut proteins using an electrophoretic separation approach showed that some digestive enzymes were absent and others were newly expressed in microbiota-free larvae. Larvae reared in antibiotic-containing refractory wheat bran diet performed poorly in comparison with controls. The addition of saligenin, the aglycone of the plant glucoside salicin, has more deleterious effects on microbiota-free larvae than on the conventionally reared larvae, suggesting a detoxifying role of midgut microbiota. Analysis of the volatile organic compounds released from the faecal pellets of the larvae shows key differences in the profiles from conventionally reared and aseptically reared larvae. Pentadecene is a semiochemical commonly found in other beetle species. Here we demonstrate the absence of pentadecene from aseptically reared larvae in contrast to its presence in conventionally reared larvae. The results are discussed in the light of the hypothesis that microbial products play subtle roles in the life of the insect, they are involved in the digestion of refractory food, detoxification of secondary plant compounds and modify the volatile profiles of the insect host.     

二化螟绒茧蜂对二化螟及其寄主植物挥发物的趋性反应

利用Y-型嗅觉仪研究了二化螟绒茧蜂Cotesia chilonis对寄主植物（水稻或茭白）、二化螟Chilo suppressalis幼虫、虫粪及虫害苗挥发物的行为反应。健康植株、二化螟幼虫和虫粪的挥发物对二化螟绒茧蜂具有显著引诱作用。在虫害苗与健康苗挥发物之间,二化螟绒茧蜂显著地偏好虫害苗,但当去除虫害苗中的幼虫和虫粪后,寄生蜂对去虫苗与机械损伤苗的选择无显著差异;在虫害苗与有虫健康苗之间,寄生蜂显著趋向虫害苗,表明虫害苗本身释放的挥发物对二化螟绒茧蜂引诱作用与机械损伤苗无显著差异,但与二化螟幼虫或虫粪挥发物之间可能具有协同增效作用。水稻苗经机械损伤或损伤后以二化螟幼虫唾液处理,其挥发物对二化螟绒茧蜂的引诱作用无显著改变。二化螟绒茧蜂对不同为害程度水稻挥发物的选择无显著差异。二化螟绒茧蜂对两种寄主植物的健康苗、虫害苗、取食两种植物的幼虫及虫粪的挥发物的选择无显著差异。结果表明,二化螟绒茧蜂栖境定位和寄主选择过程中所利用的挥发物主要来自寄主植物、二化螟幼虫和虫粪以及虫害苗与幼虫和虫粪的协同作用。     

Herbivore-induced plant volatiles mediate host selection by a root herbivore

In response to herbivore attack, plants mobilize chemical defenses and release distinct bouquets of volatiles. Aboveground herbivores are known to use changes in leaf volatile patterns to make foraging decisions, but it remains unclear whether belowground herbivores also use volatiles to select suitable host plants. We therefore investigated how above- and belowground infestation affects the performance of the root feeder Diabrotica virgifera virgifera, and whether the larvae of this specialized beetle are able to use volatile cues to assess from a distance whether a potential host plant is already under herbivore attack. Diabrotica virgifera larvae showed stronger growth on roots previously attacked by conspecific larvae, but performed more poorly on roots of plants whose leaves had been attacked by larvae of the moth Spodoptera littoralis. Fittingly, D. virgifera larvae were attracted to plants that were infested with conspecifics, whereas they avoided plants that were attacked by S. littoralis. We identified (E)-β-caryophyllene, which is induced by D. virgifera, and ethylene, which is suppressed by S. littoralis, as two signals used by D. virgifera larvae to locate plants that are most suitable for their development. Our study demonstrates that soil-dwelling insects can use herbivore-induced changes in root volatile emissions to identify suitable host plants.     

Elimination of sexual brood in the Argentine antLinepithema humile: queen effect and brood recognition

Mated queens of the antLinepithema humile (Iridomyrmex humilis Mayr) introduced into dequeened colony fragments rearing sexual brood elicited worker aggression resulting in queen larvae being bitten and eliminated. By contrast, male larvae were spared. Regarding queen brood, killing mainly concerned small and medium sized larvae. A large proportion of the large larvae escaped extermination, and prepupae and pupae were spared. These data suggest that workers were able to discriminate sex, caste and age of the brood. That a queen pheromone may be involved was shown by experiments using whole or cut corpses that were either rinsed or not rinsed in pentane. The pheromone eliciting worker aggressive behaviour was shown to act over a short distance, suggesting that it is somewhat volatile. Similarities and differences between this new queen pheromone and other known queen pheromones acting on queen production or worker attraction are discussed as well as the origin of the signals underlying the recognition of the larval classes.     

Oriented responses of carrot fly larvae, Psila rosae, to plant odours, carbon dioxide and carrot root volatiles

ABSTRACT. The responses of carrot fly larvae, Psila rosae , to host plant volatiles were studied. Two criteria for an oriented response were used: one based on whether an individual reached within 1.5 cm of the odour source and one involving analysis of the track patterns of larvae within 3 cm of the odour source. All larval instars were attracted to both host and non-host plant material, though to a lesser degree by the latter; they were also attracted to CO₂, but not to N₂. A steam volatile fraction of carrot roots, and methyl eugenol, a volatile component of this fraction, also attracted the larvae, differentially at different concentrations. The possible roles of both CO₂ and the host-specific secondary plant volatiles in the host plant finding behaviour of soil inhabiting arthropods are discussed.     

Trichomes as dangerous lollipops: Do lizards also use caterpillar body and frass odor to optimize their foraging?

When attacked by herbivores, plants produce toxic secondary metabolites that function as direct defenses, as well as indirect defenses that attract and reward predators of the offending herbivores. These indirect defenses include both nutritive rewards such as extra floral nectar, as well as informational rewards, such as the production and release of volatile compounds that betray the location of feeding herbivores to predators. Herbivory of Nicotiana attenuata by the tobacco hornworm (Manduca larvae) alters the volatile profiles of both the plant and larval headspace. Herbivory-elicited specific changes in the volatile profiles are detected by arthropod predators of Manduca larvae. The known predators that perceive volatile cues induced by Manduca herbivory of N. attenuata are insects that target Manduca at early developmental stages, when the larvae are still small; large, late-instar larvae may have outgrown these predation risks. However, here we offer evidence that branched chain aliphatic acids derived from the digestion of plant O-acyl sugars from trichomes may betray Manduca larvae to lizard predators during late developmental stages as well.     

Olfactory response of Costelytra zealandica (Coleoptera: Melolonthinae) to the roots of several pasture plants

The influence of natural plant odours on the locomotory behaviour of 3rd‐instar larvae of Costelytra zealandica (White) was studied by observing their movement in glass‐sided test chambers. Through an analysis of the paths followed by individual larvae in single‐option choice‐chamber tests it was possible to evaluate the responses of the larvae to different plant materials. The odour of fresh young perennial ryegrass (Loliumperenne) root was more attractive to the insects than was that of older plants. Larvae were also more strongly attracted to the root of lucerne (Medicago sativa), Lotus pedunculatus, red clover (Trifolium pratense), and white clover (T. repens) than to that of perennial ryegrass. The possible role of volatile chemical factors in plant resistance to grass grub attack is discussed.     

Responses of carrot fly larvae, Psila rosae, to the odorous and contact-chemostimulatory metabolites of host and non-host plants

ABSTRACT. The responses of third instar carrot fly larvae, Psila rosae , to plant metabolites were studied in a moving air olfactometer. Directed responses over distances greater than 7.5 cm were obtained to both host and non-host plant odours, to CO₂ and methyl eugenol, a volatile secondary metabolite of carrot roots. Larvae also oriented to gradients of contact-chemostimuli. The role of both olfactory and contact-chemostimuli in the host plant finding behaviour of P. rosae larvae is discussed, and observational evidence for the behavioural mechanisms involved presented.     

Oriented responses of grapevine moth larvae Lobesia botrana to volatiles from host plants and an artificial diet on a locomotion compensator

Larvae of the grapevine moth Lobesia botrana (Lepidoptera: Tortricidae) are a major pest of vine, Vitis vinifera. As larvae have limited energy reserves and are in danger of desiccation and predation an efficient response to plant volatiles that would guide them to food and shelter could be expected. The responses of starved 2nd or 3rd instar larvae to volatile emissions from their artificial diet and to single host plant volatiles were recorded on a locomotion compensator. Test products were added to an air stream passing over the 30 cm diameter servosphere. The larvae showed non-directed walks of low rectitude in the air stream alone but changed to goal-oriented upwind displacement characterised by relatively straight tracks when the odour of the artificial diet and vapours of methyl salicylate, 1-hexanol, (Z)-3-hexen-1-ol, terpinen-4-ol, 1-octen-3-ol, (E)-4,8-dimethyl-1,3,7-nonatriene and (Z)-3-hexenyl acetate were added to the air stream. This chemoanemotactic targeted displacement illustrates appetence for certain volatile cues from food by starved Lobesia larvae. Analysis of the larval behaviour indicates dose dependent responses to some of the host plant volatiles tested with a response to methyl salicylate already visible at 1 ng, the lowest source dose tested. These behavioural responses show that Lobesia larvae can efficiently locate mixtures of volatile products released by food sources as well as single volatile constituents of their host plants. Such goal-oriented responses with shorter travel time and reduced energy loss are probably of importance for larval survival as it decreases the time they are exposed to biotic and abiotic hazards.     

茉莉酸甲酯诱导邻近番茄植株抗斜纹夜蛾的浓度和距离效应

茉莉酸甲酯(Methyl jasmonate, MeJA)与植物间的防御通讯密切相关,其挥发性强,可以在空气中传播而诱导邻近植物产生防御反应,为探明MeJA诱导邻近植株抗虫性的浓度和距离效应,本研究在大棚(长4 m×宽1 m×高1 m)进口处喷以不同浓度MeJA,检测离气味源60 cm、120 cm、180 cm、240 cm和300 cm远的番茄植株叶片多酚氧化酶(Polyphenol oxidase, PPO),过氧化物酶(Peroxidase, POD)和脂氧合酶(Lipoxygenase, LOX)的活性以及斜纹夜蛾幼虫取食不同处理番茄植株叶片后的体重增长率,研究结果表明1 mmol/L和10 mmol/L MeJA挥发物可以显著诱导增强邻近番茄植株叶片PPO,POD和LOX酶的活性,以离10 mmol/L MeJA气味源60 cm处植株叶片的酶活性最高,并随着距离的增加而呈降低的趋势;取食60 cm和300 cm对照组植株叶片的斜纹夜蛾幼虫其体重增长率没有显著差异,然而1 mmol/L和10 mmol/L MeJA处理下,取食300 cm处植株叶片的斜纹夜蛾幼虫其体重增长率显...     

Birds exploit herbivore‐induced plant volatiles to locate herbivorous prey

Arthropod herbivory induces plant volatiles that can be used by natural enemies of the herbivores to find their prey. This has been studied mainly for arthropods that prey upon or parasitise herbivorous arthropods but rarely for insectivorous birds, one of the main groups of predators of herbivorous insects such as lepidopteran larvae. Here, we show that great tits (Parus major) discriminate between caterpillar‐infested and uninfested trees. Birds were attracted to infested trees, even when they could not see the larvae or their feeding damage. We furthermore show that infested and uninfested trees differ in volatile emissions and visual characteristics. Finally, we show, for the first time, that birds smell which tree is infested with their prey based on differences in volatile profiles emitted by infested and uninfested trees. Volatiles emitted by plants in response to herbivory by lepidopteran larvae thus not only attract predatory insects but also vertebrate predators.     

Neurophysiological and behavioural evidence for an olfactory function for the dorsal organ and a gustatory one for the terminal organ in Drosophila melanogaster larvae

Multicellular electrophysiological responses from the dorsal organ on the cephalic lobes of third instar Drosophila melanogaster larvae (wild-type Canton S) stimulated with a cold-trapped banana volatile extract showed that this structure has an olfactory function in the fruit fly. Responses of the dorsal organ were also recorded to constituents of the banana volatile extract as they eluted from a gas chromatographic column (GC-coupled dorsal organ electrophysiology). The active chemostimulants were identified as 2-heptanone, isoamyl alcohol, hexyl acetate, hexanol and hexyl butyrate by gas chromatography-coupled mass spectrometry. Applying the same recording system to the terminal organ sensilla, no responses were obtained to either the banana volatile bouquet or its constituents. By contrast, high frequency multicellular responses were recorded in response to touching the terminal organ with the gustatory stimuli KCl and grapefruit juice; responses were absent on similar stimulation of the dorsal organ with either NaCl or KCl. This suggests a role for olfaction by the dorsal organ and for gustation by the terminal organ in Drosophila larvae.In a 7-mm high wind tunnel with a thin 1.2% agar floor, the Drosophila larvae showed odour-conditioned upwind responses in an air stream of 0.1 m/s bearing banana volatiles. Drosophila larvae responded best to the odour of cut bananas. A 1:1 mixture of the banana odour constituents 2-heptanone and hexanol (at either 50 or 100 &mgr;g source dose each) proved as attractive as the known larval attractants propionic acid and isoamyl acetate on their own at 100 &mgr;g, whereas hexanol and 2-heptanone on their own at a 100 &mgr;g source dose were less attractive. The stronger behavioural response to the banana volatile bouquet and to the binary mixture serves to underline the multireceptor nature of the dorsal organ response to food odour in Drosophila.     

Chemical defences in leaf beetles and their larvae: The ecological, evolutionary and taxonomic significance

The chemical defences of the Chrysomelinae are reviewed. Defensive glandular secretions have evolved independently in larvae and adults, and faster than the morphology of the glands. Both characters are used in a phylogenetic study of the Chrysomelini, disclosing suprageneric affinities. First, a close relationship between the Chrysomelina and Phratora is proposed. Secondly, Leptinotarsa and Gonioctena are probably more closely related to the Chrysolinina than to the Chrysomelina and Phratora. The qualitative and quantitative composition of larval and adult secretions sometimes differs between geographically isolated populations of the same species, which are therefore probably genetically distinct. The host plant shows an influence which explains discrepancies between the current classification and the chemical nature of the secretions. Some beetles sequester secondary plant metabolites or use them as precursors for their own defences. The original biosynthetic pathway has been lost in these species. Other factors which could affect the chemical diversity of the secretions are discussed: random events such as neutral mutations and genetic drift, conditioning or adaptation of predators to specific defensive mixtures, and the diversity of potential predators. The latter could help to explain the divergence in the chemical nature of the larval and adult secretions. The volatile irritants produced by the larvae are well suited to repel small arthropods, such as ants, whereas poisons associated with aposematic coloration in the adults could be directed against small vertebrates, such as birds.     

Where to from here? The mechanisms enabling the movement of first instar caterpillars on whole plants using Helicoverpa armigera (Hübner)

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is an economically-important, polyphagous herbivore in Old World countries. The distribution of larvae within various host plants has been described, but few studies have tried to determine the behavioural mechanisms by which the given distributions arose. Our aim was to determine the mechanisms which enable larval movement on pea plants, starting with first instars. Observations and bioassays determined larval movement in response to light and angled surfaces, as well as the effect of feeding and plant volatiles on these responses. The majority (68–72%) of 1st instars were positively phototactic towards blue, green and white light and 42% towards UV light. In the dark, larvae showed negative geotaxis. The angle of their substrate also had a kinetic effect on larvae; the steeper the angle from horizontal the more larvae moved under all conditions. Phenylacetaldehyde (a flower volatile) suppressed larval movement except at 90°. (Z)-3-Hexenyl acetate (a green leaf volatile) reversed the direction of movement at the flattest angle. Feeding lessened the probability of moving. We suggest that phototaxis and geotaxis are behaviours common to larval lepidopterans (caterpillars), and that these basic behaviours are modulated by environmental, larval, and plant factors to give observed distributions. Using a multinomial model approach, we created a flow chart to qualitatively and quantitatively represent the decision-making process of first instar H. armigera in response to the factors influencing movement.     

Chemical composition and anti-mosquito potential of rhizome extract and volatile oil derived from Curcuma aromatica against Aedes aegypti (Diptera: Culicidae).

Crude rhizome extracts and volatile oils of Curcuma aromatica were evaluated for chemical composition and anti-mosquito potential, including larvicidal, adulticidal, and repellent activities against the Aedes aegypti mosquito. Chemical identification achieved by GC/MS analysis revealed that xanthorrhizol, 1H-3a, 7-methanoazulene and curcumene at 35.08 and 13.65%, 21.81 and 30.02%, and 13.75 and 25.71%, were the main constituents in hexane extracts and volatile oils, respectively. Volatile oil of Cu. aromatica possessed a significantly higher larvicidal activity against the 4th instar larvae of Ae. aegypti than that of hexane extracts, with LC50 values of 36.30 and 57.15 ppm, respectively. In testing for adulticidal activity, on the other hand, hexane-extracted Cu. aromatica (LC50: 1.60 microg/mg female) was found to be slightly more effective against female Ae. aegypti than volatile oil (LC50: 2.86 microg/mg female). However, the repellency of these two products against Ae. aegypti adult females differed significantly. The hexane-extracted Cu. aromatica, with a median complete protection time of 1 h (range = 1-1.5 h) when applied at a concentration of 25%, appeared to have significantly higher repellency than that of distillate oil (0.5 h, range = 0-0.5 h). The different results obtained from both products of Cu. aromatica were probably due to variety in quantity and type of active ingredients as well as the biological and physiological characteristics that differed between both developmental stages of mosquitoes, larvae, and adults.     

Methyl palmitate does not elicit invasion of honeybee brood cells by Varroa mites

Since Varroa mites decide at some distance from the larva whether to stay on a bee or invade a cell, they may well use a volatile chemical to select a brood cell. A few aliphatic esters, predominantly methyl palmitate, have been claimed to be this volatile signal for the mites for two reasons. The mites respond to the esters in an olfactometer (Le Conte et al., 1989), and the levels of the esters in worker and drone larvae may explain that drone cells are attractive during a longer period and are invaded more frequently than worker cells (Trouiller et al., 1992). However, invasion itsclf appeared to be unaffected by application of methyl palmitate to brood cells. In addition, analysis of the volatiles emanating from attractive brood cells showed hundreds of components in the volatile blend, but in only 2 of 17 analyses a trace of methyl palmitate was found. Hence, there is no reason to believe that methyl palmitate is used as a signal for invasion by the mites.     

Feeding by Hessian fly [Mayetiola destructor (Say)] larvae does not induce plant indirect defences

Abstract.  1. Recent research has addressed the function of herbivore-induced plant volatiles in attracting natural enemies of feeding herbivores. While many types of insect herbivory appear to elicit volatile responses, those triggered by gall insects have received little attention. Previous work indicates that at least one gall insect species induces changes in host-plant volatiles, but no other studies appear to have addressed whether gall insects trigger plant indirect defences.
2. The volatile responses of wheat to feeding by larvae of the Hessian fly Mayetiola destructor (Say) (Diptera: Cecidomyiidae) were studied to further explore indirect responses of plants to feeding by gall insects. This specialist gall midge species did not elicit a detectable volatile response from wheat plants, whereas a generalist caterpillar triggered volatile release. Moreover, Hessian fly feeding altered volatile responses to subsequent caterpillar herbivory.
3. These results suggest that Hessian fly larvae exert a degree of control over the defensive responses of their host plants and offer insight into plant-gall insect interactions. Also, the failure of Hessian fly larvae to elicit an indirect defensive response from their host plants may help explain why natural enemies, which often rely on induced volatile cues, fail to inflict significant mortality on M. destructor populations in the field.     

The effect of lactic acid and the volatile fatty acids on the aggregation behavior of Periplaneta americana (L.)

Young larvae of Periplaneta americana (L.) are attracted to filter paper wetted with 10(-2)-10(-3)% solutions of butyric acid. Other volatile fatty acids and lactic acid, as well as other organic and inorganic acids, are repellent to larvae at high concentrations, and ineffective at low.