(2R,5S)-Theaspirane Identified as the Kairomone for the Banana Weevil, Cosmopolites sordidus, from Attractive Senesced Leaves of the Host Banana, Musa spp.

The principal active component produced by highly attractive senesced host banana leaves, Musa spp., for the banana weevil, Cosmopolites sordidus, is shown by coupled gas chromatography-electroantennography (GC-EAG), coupled GC-mass spectrometry (GC-MS), chemical synthesis and coupled enantioselective (chiral) GC-EAG to be (2R,5S)-theaspirane. In laboratory behaviour tests, the synthetic compound is as attractive as natural host leaf material and presents a new opportunity for pest control.

The sex pheromones of mealy plum (Hyalopterus pruni) and leaf-curl plum (Brachycaudus helichrysi) aphids: identification and field trapping of male and gynoparous aphids in prune orchards.

Mealy plum, Hyalopterus pruni, and leaf-curl plum, Brachycaudus helichrysi, aphids are the primary arthropod pests in orchards that produce dried plums (i.e., prunes). The sexual stage of their respective lifecycles occurs on prune trees in the fall, during which time males respond to sex pheromones produced by oviparous females. Air-entrainment collections confirmed that oviparous H. pruni and B. helichrysi emitted combinations of (4aS, 7S, 7aR)-nepetalactone and (1R, 4aS, 7S, 7aR)-nepetalactol. The responses of H. pruni and B. helichrysi to these compounds in ratios of 1:0, 0:1, 1:1, 2.6:1, 3.4:1, 5:1, 7:1, and 0:0 (no-pheromone control) using water traps were determined in field experiments conducted in prune orchards during the fall. The greatest number of male H. pruni was caught in traps releasing a 1:1 ratio of (4aS, 7S, 7aR)-nepetalactone and (1R, 4aS, 7S, 7aR)-nepetalactol, while male B. helichrysi were caught in similar numbers in traps releasing any of the two-component ratios tested. There was no evidence that any of the pheromone treatments influenced trap catches of gynoparae of either species. Results suggest that addition of sex pheromone lures increases trap catches of male H. pruni and B. helichrysi, and that this approach may improve monitoring and management of these pests in prune orchards. Knowledge gained from this study contributes to the understanding of the ecology of insect pests in prune orchards.

Use of honey bees (Apis mellifera L.) to detect the presence of Mediterranean fruit fly (Ceratitis capitata Wiedemann) larvae in Valencia oranges.

BACKGROUND: When fruit deteriorates a characteristic profile of volatile chemicals is produced that is different from that produced by healthy fruits. The identification of such chemicals allows the possibility of monitoring the fruit for early signs of deterioration with biological sensors. The use of honey bees and other insects as biological sensors is well known. This study aimed to identify the volatiles produced by oranges infested with larvae of the Mediterranean fruit fly and to test the ability of honey bees, conditioned to this volatile chemical profile, to detect such oranges. RESULTS: Seventeen compounds that were present in higher concentrations in the volatile profiles of infested oranges than in those of insect-free fruits were mixed at the same relative concentrations as those in the collected volatiles of infested oranges. The synthetic mixture was used to train honey bees by classical Pavlovian conditioning and subsequent tests showed that they were then able to discriminate between medfly-infested and uninfested oranges. CONCLUSION: This study demonstrates an innovative way of detecting, at an early stage, the symptoms of damage to oranges by the Mediterranean fruit fly.

cis-Jasmone induces Arabidopsis genes that affect the chemical ecology of multitrophic interactions with aphids and their parasitoids.

It is of adaptive value for a plant to prepare its defenses when a threat is detected, and certain plant volatiles associated with insect damage, such as cis-jasmone (CJ), are known to switch-on defense metabolism. We used aphid and aphid parasitoid responses to Arabidopsis thaliana as a model system for studying gene expression and defense chemistry and its impact at different trophic levels. Differential responses to volatiles of induced Arabidopsis occurred for specialist and generalist insects: the generalist aphid, Myzus persicae, was repelled, whereas the specialist, Lipaphis erysimi, was attracted; the generalist aphid parasitoid Aphidius ervi was attracted, but the specialist parasitoid Diaeretiella rapae was not affected. A. ervi also spent longer foraging on induced plants than on untreated ones. Transcriptomic analyses of CJ-induced Arabidopsis plants revealed that a limited number of genes, including a gene for a cytochrome P450, CYP81D11, were strongly up-regulated in the treated plants. We examined transgenic Arabidopsis lines constitutively overexpressing this gene in bioassays and found insect responses similar to those obtained for wild-type plants induced with CJ, indicating the importance of this gene in the CJ-activated defense response. Genes involved in glucosinolate biosynthesis and catabolism are unaffected by CJ and, because these genes relate to interactions with herbivores and parasitoids specific to this family of plants (Brassicaceae), this finding may explain the differences in behavioral response of specialist and generalist insects.

Vicia faba-Lygus rugulipennis interactions: induced plant volatiles and sex pheromone enhancement.

The profiles of volatile chemicals emitted by Vicia faba plants damaged by Lygus rugulipennis feeding, and by feeding plus oviposition, were shown to be quantitatively different from those released by undamaged plants. Samples of volatile chemicals collected from healthy plants, plants damaged by males as a consequence of feeding, plants damaged by females as a consequence of feeding and oviposition, plants damaged by feeding with mated males still present, and plants damaged by feeding and oviposition with gravid females still present, showed significant differences in the emission of hexyl acetate, (Z)-beta-ocimene, (E)-beta-ocimene, (E)-beta-caryophyllene, and methyl salicylate. In particular, treatments with mated females present on plants had a significant increase in emission levels of the above compounds, possibly due to eggs laid within plant tissues or active feeding, compared with undamaged plants and plants damaged by males feeding, with or without insects still present. Furthermore, the pheromonal blend released by mated L. rugulipennis females, mainly comprising hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-4-oxo-2-hexenal, was enhanced when females were active on broad bean plants, whereas such an increase was not observed in males. Both sexes gave electroantennogram responses to green leaf volatiles from undamaged plants and to methyl salicylate and (E)-beta-caryophyllene emitted by Lygus-damaged plants, suggesting that these compounds may be involved in colonization of host plants by L. rugulipennis. In addition, mated males and females were responsive to hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-4-oxo-2-hexenal released by mated females on V. faba, indicating that these substances could have a dual function as a possible aggregation pheromone in female-female communication, and as a sex pheromone in female-male communication.

Attractiveness of host banana leaf materials to the banana weevil, Cosmopolites sordidus in Ghana for development of field management strategies.

BACKGROUND: The banana weevil, Cosmopolites sordidus, has been frequently cited as the most challenging constraint to banana and plantain production, particularly in small-scale (smallholder) farming. For the development of a new, low-cost weevil management technology based on attractive host plant material, we previously identified (2R,5S)-theaspirane as the active component of attractive senesced banana leaves. In this new study, we used behavioural (olfactometer) bioassays with adult weevils to compare the attractiveness of four different developmental stages of banana leaves, i.e. unfolding (pale green), matured green (deep green), matured yellowing and senesced, to determine which leaf developmental stage would be most appropriate for use in weevil management. We also investigated the attractiveness of senesced leaf extracts prepared using different solvents to determine which solvent would be most appropriate for local production of leaf extracts. Coupled gas chromatography-electroantennography (GC-EAG) was then used with adult weevils to confirm the presence of (2R,5S)-theaspirane in attractive leaf extracts. RESULTS: Of the leaf materials tested, only the odour of senesced leaf material was significantly attractive to adult weevils (P < 0.005). Furthermore, an extract of senesced material prepared using palm wine alcohol was significantly attractive (P < 0.05). Using coupled GC-EAG with weevil antennae, (2R,5S)-theaspirane was identified as a minor component with strong EAG activity within the palm wine alcohol extract. CONCLUSION: The results suggest that palm wine alcohol extracts of senesced banana leaf material could be used to lure adult C. sordidus to traps in the field, as part of an ethnobotanical-based approach for C. sordidus management on smallholder farms. © 2018 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Antiectoparasitic activity of the gum resin, gum haggar, from the East African plant, Commiphora holtziana.

The mechanism of ixodid tick (Acari: Ixodidae) repellency by gum haggar, a resin produced by Commiphora holtziana (Burseraceae), was investigated by evaluating activity against the cattle tick, Boophilus microplus. In an arena bioassay, a hexane extract of the resin of C. holtziana exhibited a repellent effect lasting up to 5h. The hydrocarbon fraction of the resin extract was shown to account for the repellent activity, and was analysed by coupled gas chromatography-mass spectrometry (GC-MS). Major sesquiterpene hydrocarbons were tentatively identified as germacrene-D, delta-elemene and beta-bourbonene. The identity and stereochemistry of the former compound was confirmed as the (+)-isomer by peak enhancement using enantioselective GC, whereas the latter 2 compounds, which are most likely degradation products of germacrene-type precursors, were identified through isolation by preparative gas chromatography followed by microprobe-NMR spectroscopy. GC comparison of gum haggar with another resin, C. myrrha, which was inactive in the tick bioassay, showed that the latter contained much lower levels of these hydrocarbons. To assess the suitability of the gum haggar resin as a general acarine repellent, further tests were made on a major acarine pest of European and US animal husbandry systems, the red poultry mite, Dermanyssus gallinae (Acari: Dermanyssidae). Gum haggar extract, and the isolated hydrocarbon fraction, showed strong repellent effects in an olfactometer assay, and again gum myrrh showed no effect. These findings provide a scientific basis for the observed anti-tick properties of gum haggar, and demonstrate the potential for its development as a general acarine repellent for use in animal husbandry systems.

Developments in aspects of ecological phytochemistry: the role of cis-jasmone in inducible defence systems in plants.

The challenges and opportunities for protecting agricultural production of food and other materials will be met through exploiting the induction of defence pathways in plants to control pests, diseases and weeds. These approaches will involve processes that can be activated by application of natural products, patented in terms of this use, to "switch on" defence pathways. Already, a number of secondary metabolite defence compounds are known for which the pathways are conveniently clustered genomically, e.g. the benzoxazinoids (hydroxamic acids) and the avenacins. For the former, it is shown that the small molecular weight lipophilic activator cis-jasmone can induce production of these compounds and certain genes within the pathway. Numerous groups around the world work on inducible defence systems. The science is rapidly expanding and involves studying the interacting components of defence pathways and the switching mechanisms activated by small molecular weight lipophilic compounds. Examples are described of how plant breeding can exploit these systems and how heterologous gene expression will eventually give rise to a new range of GM crops for food and energy, without the need for external application of synthetic pesticides.

Novel olfactory ligands via terpene synthases.

A synthetic biology approach to the rational design of analogues of olfactory ligands by providing unnatural substrates for the enzyme synthesising (S)-germacrene D, an olfactory ligand acting as a plant derived insect repellent, to produce novel ligands is described as a viable alternative to largely unsuccessful ligand docking studies. (S)-14,15-Dimethylgermacrene D shows an unexpected reversal in behavioural activity.

Insecticidal 2-hydroxy-3-alkyl-1,4-naphthoquinones: correlation of inhibition of ubiquinol cytochrome c oxidoreductase (complex III) with insecticidal activity.

The insecticidal and in vitro activities of four homologous series of 2-hydroxy and acetoxy-3-substituted-1,4-naphthoquinones have been measured and correlated with their (Log) octanol/water partition coefficients (Log Ko/w). In vitro activity against mitochondrial complex III was only exhibited by 2-hydroxy-3-alkyl-1,4-naphthoquinones, indicating that the 2-acetoxy compounds act as proinsecticides. Good correlation was observed between in vivo activity against the two-spotted spider mite, Tetranychus urticae and inhibition of complex III isolated from blowfly flight muscle. Both hydroxy and acetoxy analogues of individual compounds exhibited similar levels of in vivo activity with optimum activity for analogues with Log Ko/w values of 7-8. In contrast, the acetoxy derivatives showed superior in vivo activity against the tobacco whitefly, Bemisia tabaci. Complex III isolated from whitefly was optimally inhibited by hydroxy analogues with lower Log Ko/w values (6.0-6.5) and was also more sensitive than the blowfly enzyme to all the compounds tested.

Emerging roles in plant defense for cis-jasmone-induced cytochrome P450 CYP81D11.

cis-Jasmone is a volatile organic compound emitted constitutively by flowers or leaves of several plant species where it acts as an attractant for pollinators and as a chemical cue for host localisation (or avoidance) for insects. ( 1-3) It is also released by some plant species after feeding damage inflicted by herbivorous insects and in this case might serve as a chemical cue for parasitoids to guide them to their prey (so called "indirect defense"). ( 4,5) Moreover, we have recently shown that plants can perceive cis-jasmone and that it acts as a signaling molecule in A. thaliana, inducing a discrete and distinctive suite of genes, of which a large subset is putatively involved in metabolism and defense responses. ( 6) Cytochrome P450s feature prominently in these functional subsets and of these the highest fold change upon cis-jasmone treatment occurred with the cytochrome CYP81D11 (At3g28740). ( 6) Hence this gene was chosen for a more thorough analysis of the potential biological relevance of the cis-jasmone induced defense response. Although the precise function of CYP81D11 remains to be determined, we could previously demonstrate its involvement in the indirect defense response in Arabidopsis, as plants exposed to cis-jasmone ceased to be attractive to the aphid parasitoid Aphidius ervi when this P450 was inactivated by T-DNA insertion mutagenesis. ( 6) Here we report additional experiments which give further support to a role of CYP81D11 in the direct or indirect defense response of A. thaliana.

Isoschaftoside, a C-glycosylflavonoid from Desmodium uncinatum root exudate, is an allelochemical against the development of Striga.

In East African small-holder farming of maize, the cattle forage legume, Desmodium uncinatum is used as an intercrop due to its allelopathic inhibition of parasitism by Striga hermonthica, an obligate parasitic weed that can devastate the maize crop. Bioassay-guided fractionation of the root extract of D. uncinatum revealed isoschaftoside to be the main compound in the most potent fraction inhibiting growth of germinated S. hermonthica radicles. Bioassays repeated with isoschaftoside isolated from a different plant source, Passiflora incarnata, proved it to be a biologically active component. Analysis of the root exudates produced by hydroponically grown D. uncinatum showed isoschaftoside to be present in the hydroponic media at biologically active concentrations of 10-100 nM.

New genetic opportunities from legume intercrops for controlling Striga spp. parasitic weeds.

In smallholder farming in East Africa, intercropping of maize with the cattle forage legume, Desmodium uncinatum Jacq., prevents parasitism by Striga hermonthica (Del.) Benth. (witchweed) through an allelopathic mechanism. Isoschaftoside, a di-C-glycosylflavone, isolated from the root extract and root exudate of Desmodium, interferes with in vitro radicle development of germinated Striga. The biosynthetic pathway of this class of compound is already mostly present in edible legumes and in cereals, so characterisation of the enzyme and genes that control C-glycosylflavone biosynthesis has the potential to create this protection mechanism in other agriculturally important plants.

Aphid alarm pheromone produced by transgenic plants affects aphid and parasitoid behavior.

The alarm pheromone for many species of aphids, which causes dispersion in response to attack by predators or parasitoids, consists of the sesquiterpene (E)-beta-farnesene (Ebetaf). We used high levels of expression in Arabidopsis thaliana plants of an Ebetaf synthase gene cloned from Mentha x piperita to cause emission of pure Ebetaf. These plants elicited potent effects on behavior of the aphid Myzus persicae (alarm and repellent responses) and its parasitoid Diaeretiella rapae (an arrestant response). Here, we report the transformation of a plant to produce an insect pheromone and demonstrate that the resulting emission affects behavioral responses at two trophic levels.