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PubMed2005

Evaluating the induced-odour emission of a Bt maize and its attractiveness to parasitic wasps.

The current discussion on the safety of transgenic crops includes their effects on beneficial insects, such as parasitoids and predators of pest insects. One important plant trait to consider in this context is the emission of volatiles in response to herbivory. Natural enemies use the odours that result from these emissions as cues to locate their herbivorous prey and any significant change in these plant-provided signals may disrupt their search efficiency. There is a need for practical and reliable methods to evaluate transgenic crops for this and other important plant traits. Moreover, it is imperative that such evaluations are done in the context of variability for these traits among conventional genotypes of a crop. For maize and the induction of volatile emissions by caterpillar feeding this variability is known and realistic comparisons can therefore be made. Here we used a six-arm olfactometer that permits the simultaneous collection of volatiles emitted by multiple plants and testing of their attractiveness to insects. With this apparatus we measured the induced odour emissions of Bt maize (Bt11, N4640Bt) and its near-isogenic line (N4640) and the attractiveness of these odours to Cotesia marginiventris and Microplitis rufiventris, two important larval parasitoids of common lepidopteran pests. Both parasitoid species were strongly attracted to induced maize odour and neither wasp distinguished between the odours of the transgenic and the isogenic line. Also wasps that had previously experienced one of the odours during a successful oviposition divided their choices equally between the two odours. However, chemical analyses of collected odours revealed significant quantitative differences. The same 11 compounds dominated the blends of both genotypes, but the isogenic line released a larger amount of most of these. These differences may be due to altered resource allocation in the transgenic line, but it had no measurable effect on the wasps' behaviour. All compounds identified here had been previously reported for maize and the differential quantities in which they were released fall well within the range of variability observed for other maize genotypes.

Animals↗
PubMed2006

CFTR surface expression and chloride currents are decreased by inhibitors of N-WASP and actin polymerization.

The cystic fibrosis transmembrane conductance regulator (CFTR) undergoes rapid turnover at the plasma membrane in various cell types. The ubiquitously expressed N-WASP promotes actin polymerization and regulates endocytic trafficking of other proteins in response to signaling molecules such as Rho-GTPases. In the present study we investigated the effects of wiskostatin, an N-WASP inhibitor, on the surface expression and activity of CFTR. We demonstrate, using surface biotinylation methods, that the steady-state surface CFTR pool in stably transfected BHK cells was dramatically decreased following wiskostatin treatment with a corresponding increase in the amount of intracellular CFTR. Similar effects were observed for latrunculin B, a specific actin-disrupting reagent. Both reagents strongly inhibited macroscopic CFTR-mediated Cl(-) currents in two cell types including HT29-Cl19A colonic epithelial cells. As previously reported, CFTR internalization from the cell surface was strongly inhibited by a cyclic-AMP cocktail. This effect of cyclic-AMP was only partially blunted in the presence of wiskostatin, which raises the possibility that these two factors modulate different steps in CFTR traffic. In kinetic studies wiskostatin appeared to accelerate the initial rate of CFTR endocytosis as well as inhibit its recycling back to the cell surface over longer time periods. Our studies implicate a role for N-WASP-mediated actin polymerization in regulating CFTR surface expression and channel activity.

Actins↗
PubMed2005

Caenorhabditis elegans WASP-interacting protein homologue WIP-1 is involved in morphogenesis through maintenance of WSP-1 protein levels.

Mammalian WASP and N-WASP are involved in reorganization of the actin cytoskeleton through activation of the Arp2/3 complex and in regulation of cell motility or cell shape changes. In the present study, we identified WASP-interacting protein homologue (WIP)-1 in Caenorhabditis elegans. WIP-1 contains the domains and sequences conserved among mammalian WIP family proteins. Yeast two-hybrid analysis detected a physical interaction between WIP-1 and WSP-1, the sole homologue of WASP/N-WASP in C. elegans. Western analysis of embryo lysates showed that RNA interference (RNAi) treatment for wip-1 decreased levels of WSP-1 protein, and wsp-1(RNAi) treatment decreased levels of WIP-1 protein. However, wsp-1 mRNA levels were not decreased in wip-1(RNAi)-treated embryos, and wip-1 mRNA levels were not decreased in wsp-1(RNAi)-treated embryos. Furthermore, disruption of WIP-1 by RNAi resulted in embryonic lethality with morphologic defects in hypodermal cell migration, a process known as ventral enclosure. This phenotype was similar to that observed in RNAi experiments for wsp-1. Immunostaining showed that WIP-1 was expressed by migrating hypodermal cells, as was WSP-1. This expression during ventral enclosure was reduced in wip-1(RNAi)-treated embryos and wsp-1(RNAi)-treated embryos. Our results suggest that C. elegans WIP-1 may function in hypodermal cell migration during ventral enclosure by maintaining levels of WSP-1.

Amino Acid Sequence↗
PubMed2004

EspFU is a translocated EHEC effector that interacts with Tir and N-WASP and promotes Nck-independent actin assembly.

Several microbial pathogens including enteropathogenic E. coli (EPEC) exploit mammalian tyrosine-kinase signaling cascades to recruit Nck adaptor proteins and activate N-WASP-Arp2/3-mediated actin assembly. To promote localized actin "pedestal formation," EPEC translocates the bacterial effector protein Tir into the plasma membrane, where it is tyrosine-phosphorylated and binds Nck. Enterohemorrhagic E. coli (EHEC) also generates Tir-dependent pedestals, but in the absence of phosphotyrosines and Nck recruitment. To identify additional EHEC effectors that stimulate phosphotyrosine-independent actin assembly, we systematically generated EHEC mutants containing specific deletions in putative pathogenicity-islands. Among 0.33 Mb of deleted sequences, only one ORF was critical for pedestal formation. It lies within prophage-U, and encodes a protein similar to the known effector EspF. This proline-rich protein, EspFU, is the only EHEC effector of actin assembly absent from EPEC. Whereas EHEC Tir cannot efficiently recruit N-WASP or trigger actin polymerization, EspFU associates with Tir, binds N-WASP, and potently stimulates Nck-independent actin assembly.

Actins↗
PubMed2001

Haemocyte changes in resistant and susceptible strains of D. melanogaster caused by virulent and avirulent strains of the parasitic wasp Leptopilina boulardi.

Two strains of Drosophila melanogaster (resistant and susceptible) were parasitized by a virulent or avirulent strain of the parasitoid wasp Leptopilina boulardi. The success of encapsulation depends on both the genetic status of the host strain and the genetic status of the parasitoid strain: the immune cellular reaction (capsule) is observed only with the resistant strain-avirulent strain combination. The total numbers of host haemocytes increased in all 4 combinations, suggesting that an immune reaction was triggered in all hosts. Resistant host larvae infected with the virulent or avirulent strains of parasitoid wasp had slightly more haemocytes per mm(3) than did susceptible host larvae at the beginning of the reaction (less than 15 h post-parasitization). This difference disappeared later. Only the virulent parasitoid strain caused the production of a high percentage of altered lamellocytes (from a discoid shape to a bipolar shape), half the total number of lamellocytes are altered. This suggests that the alteration of lamellocyte shape alone is not sufficient to explain the lack of capsule formation seen in resistant hosts parasitized by the virulent strain. Lastly, there were very few altered lamellocytes in resistant or susceptible hosts parasitized by the avirulent parasitoid strain, two combinations in which no capsule was formed. As is now established for Drosophila-parasitoid interactions, virus-like particles contained in the long gland of the female wasp affect the morphology of the lamellocytes. The results presented here are further proof of the action (direct or indirect) of virus like particles of the virulent strain on lamellocytes.

Journal Article↗
PubMed2001

Two distinct reproductive strategies are correlated with an ovarian phenotype in co-existing parthenogenetic strains of a parasitic wasp.

The question whether different organisms are able to compete for the same resource is of fundamental importance to evolutionary biology. Sympatric co-existence of similar species on a single resource has long been claimed to be unstable. However, indirect evidence suggests that parasitic wasps exhibit evolutionarily stable mixtures of life-history strategies. Here we describe genetically distinct strains of a parthenogenetic wasp Venturia canescens, with different ovarian phenotypes that affect egg numbers in oviducts. Wasp females with large egg load search for caterpillars and deposit eggs immediately after host encounter, whereas females with fewer eggs delay parasitism. Since the outcome of interlarval competition within super-parasitized caterpillars depends on the age distribution of competing larvae, the two egg deposition strategies may co-exist under conditions that favor super-parasitism.

Journal Article↗
PubMed2002

Larvae of the ectoparasitic wasp, Eulophus pennicornis, release factors which adversely affect haemocytes of their host, Lacanobia oleracea.

When larvae of the ectoparasitic wasp Eulophus pennicornis were incubated for 4 h on balls of cotton wool soaked in tissue culture medium (TC-100), they released a variety of factors. Subsequent incubation of these larval wasp secretions with monolayers of haemocytes from their host, Lacanobia oleracea, demonstrated that they adversely affect haemocyte morphology, behaviour and viability. For instance, when monolayers of haemocytes were incubated for 18 h in TC-100, approximately 73% of the cells present, attached firmly to and spread over the tissue culture surface by extending pseudopods. By contrast, when incubated in TC-100 containing larval wasp secretions, only about 27% of the haemocytes present remained attached to the tissue culture surface after washing. The majority of these had a rounded configuration and neither spread nor extended pseudopods. Furthermore, viability assays indicated that approximately 36% of the attached haemocytes were dead, as opposed to 11-12% in the controls. The E. pennicornis secretions also significantly reduced the ability of L. oleracea haemocytes to move across the surface of the slide and form clumps (p</=0.0005) and to phagocytose FITC-labelled Escherichia coli in vitro (p</=0.0005). These results indicate that secretions from E. pennicornis larvae contain an anti-haemocyte factor(s) that can kill and/or alter the behaviour of host haemocytes. As a result, the ability of the haemocytes to execute important immune responses is compromised. Preliminary data suggest that the active molecules are proteins, and that their mechanism of action may involve inhibition of polymerization and/or disorganization of the haemocyte cytoskeleton.

Journal Article↗
PubMed1997

Effects of Parasitism by the Braconid Wasp Cotesia congregata on Metabolic Rate in Host Larvae of the Tobacco Hornworm, Manduca sexta.

We examined growth rates, gas exchange patterns and energy metabolism of tobacco hornworm (Manduca sexta) larvae parasitized by the braconid wasp Cotesia congragata. Larvae parasitized at the beginning of the fourth-instar had reduced growth compared to unparasitized larvae of the same age and short-term differences in metabolism (measured as rates of CO(2) production, Vdot; CO(2)) were apparent almost immediately after wasp oviposition. However, over the growth period between parasitization and the last part of the fifth-instar, there was no significant difference between parasitized and unparasitized hosts as seen in the relationship between mass and Vdot; CO(2). One day prior to parasitoid emergence, host larvae stopped eating, ceased spontaneous locomotor activity and showed a dramatic decline in metabolism. The 60% decline of Vdot; CO(2) at this time is consistent with lack of specific dynamic action because the animals were not feeding. Gas exchange became highly cyclical on the day of parasitoid emergence, but the cause and significance of this phenomenon, which disappeared by the third day following emergence, are not clear. This pattern of cycling was not induced by starving nonparasitized larvae for 6days, nor by immobilizing nonparasitized larvae with tetrodotoxin. Ecdysteroid levels in the host's hemolymph significantly increased on the day when parasitoids completed their L2-L3 molt and began emerging, but not during the wasps' L1-L2 molt which occurred a few days earlier. Contrary to our initial expectation that hemolymph ecdysteroid titers might be linked to alterations in the host's metabolic rate, we observed no such correlation.

Journal Article↗
PubMed1997

Caste formation in the polyembryonic wasp Copidosoma floridanum (Hymenoptera: Encyrtidae): in vivo and in vitro analysis.

The polyembryonic wasp Copidosoma floridanum produces two morphologically distinct types of larvae in its host Trichoplusia ni. Reproductive larvae consume the host, pupate, and form adult wasps, whereas precocious larvae manipulate the sex ratios of the reproductive caste and defend the brood against interspecific competitors. The previous study indicated that morphogenesis of the reproductive caste was associated with a 9-day competency period, and that ecdysteroids of host origin were required for completion of embryogenesis. Here we investigated whether factors associated with the host environment mediate morphogenesis of precocious larvae and caste determination. Embryogenesis of precocious larvae was found to be synchronized with specific stages of the host first-fourth instars. However, development of precocious larvae did not depend on environmental factors specifically associated with these host stages. Elevation of the host juvenoid titer using the analogue methoprene induced T. ni to undergo a supernumerary sixth instar, but did not alter the proportion of wasp embryos that developed into precocious and reproductive larvae. In contrast, embryos competent to initiate morphogenesis developed into precocious larvae when transplanted into novel host stages such as pupae. Development of precocious larvae was arrested by ablation of the host's source of ecdysteroids, but could be rescued dose-dependently by injection of 20-hydroxyecdysone. In vitro rearing studies confirmed that completion of embryogenesis of the precocious caste required an exogenous pulse of 20-hydroxyecdysone. Combined with previous studies, our results indicate that embryos forming precocious and reproductive larvae acquire the competence to undergo morphogenesis at different times. However, we find no evidence to suggest that caste determination is mediated by environmental factors associated with a specific stage of the host.

Journal Article↗
PubMed1999

Free serosal cells originating from the embryo of the wasp Diadromus pulchellus in the pupal body of parasitized leek-moth, Acrolepiosis assectella. Are these cells teratocyte-like?

In braconid species, teratocytes are derived from a serosal cell membrane which envelops the developing parasitoid embryo. On hatching, this membrane dissociates into individual cells, the teratocytes, which then circulate in the haemolymph of the host. We describe herein such a membrane, surrounding the embryo in eggs of the ichneumonid parasitoid wasp, Diadromus pulchellus. This membrane consisted of a single sheet of tightly packed cells with large 12+/-1.4 &mgr;m nuclei. These cells were released after hatching in vitro and cells of the same size were detected in vivo, in the vicinity of the D. pulchellus embryo. The number of nuclei detected suggests that the serosal membrane consists of about 450+/-150 cells. These cells did not grow after hatching of the parasitoid egg in the parasitized host, Acrolepiosis assectella, during the development of the parasitoid wasp larva. Southern blot experiments, using D. pulchellus satellite DNA or the ribosomal genes as probes, showed that free-living floating cells of wasp origin were present in the body of the parasitized host. This is the first time that free-floating teratocyte-like cells have been described in species of the Ichneumonidae.

Journal Article↗
PubMed2000

Parasitization of Lacanobia oleracea (Lepidoptera) by the ectoparasitic wasp, Eulophus pennicornis, suppresses haemocyte-mediated recognition of non-self and phagocytosis.

Although many endoparasitic wasps suppress the haemocyte-mediated immune defences of their insect hosts, the effects of ectoparasitoids are virtually unknown. In view of this, a study has been made of the ectoparasitic wasp, Eulophus pennicornis, and its host, the tomato moth, Lacanobia oleracea. For unparasitized insects, in vitro assays indicated that less than 3.0% of L. oleracea haemocytes on a monolayer formed rosettes with yeast cells or fresh rabbit erythrocytes (rbc), and virtually no phagocytosis of these particles occurred. In addition, although fixed rbc formed rosettes with 51.21% of haemocytes, only about 3.0% of the haemocytes ingested one or more of these particles. In contrast to this, B. cereus and E. coli were readily phagocytosed by 14.75% and 53.70% of haemocytes, respectively. These results indicate that L. oleracea haemocytes can recognise different types of non-self particles and demonstrate that ingestion does not necessarily follow attachment. When rosetting and phagocytosis assays were performed with fixed rbc and FITC-labelled E. coli, and haemocytes from starved L. oleracea, PBS injected L. oleracea, and experimentally envenomated insects on day five of treatment, there was no significant difference in the percentage of rosetting or phagocytosis occurring. When haemocytes from parasitized insects on day five of treatment were utilised, however, rosetting and phagocytosis were reduced by 31.41% and 34.94%, respectively. Thus, the effects of parasitization and experimental envenomation are not the same. In addition, suppression of host haemocyte-mediated recognition and phagocytosis was not a secondary effect of nutritional deprivation and was not due to ectoparasitoid venom components, rather it was a direct result of parasitization of L. oleracea by E. pennicornis. The putative nature and source of the immunosuppressive factor(s) involved is discussed with reference to those produced by endoparasitic wasps.

Journal Article↗
PubMed2000

Use of Dufour's gland secretion in nest defence and brood nutrition by hover wasps (Hymenoptera, Stenogastrinae).

Social wasps of the subfamily Stenogastrinae produce an abdominal secretion that is used in two distinct biological contexts. First, the secretion plays an important role in larval nutrition where it serves as a substrate in which food is placed by the adults for eventual consumption by the larvae. Second, in several species, females apply the same secretion to the substrate on which their nests are constructed, where it constitutes a sticky barrier that defends the immature brood from predation by ants. This paper describes for the first time ant guard construction behaviour of three species of stenogastrine wasps belonging to the genera Eustenogaster and Liostenogaster. The identification of compounds making up these secretions was also performed by gas chromatography-mass spectrometry. Ant guards and brood secretions were similar, with saturated and unsaturated long chain hydrocarbons and alcohols as major components. We further confirm that the glandular source of abdominal secretion is the Dufour's gland. This gland contains the same hydrocarbons, and in the same proportions as ant guards and brood secretion. We discuss the fundamental importance of Dufour's gland secretion in the social life of these wasps by comparing species with and without ant guards within the subfamily.

Journal Article↗
PubMed2001

Augmentative releases of parasitoid wasps in stored wheat reduces insect fragments in flour.

Field studies were conducted to assess the effectiveness of the parasitoid wasp, Theocolax elegans, for reducing insect fragments in flour by suppressing populations of Rhyzopertha dominica in six bins, each containing 27 tonnes of wheat. Beetles were released into all the six bins at monthly intervals for 3 months. Parasitoid wasps were released into three of the bins, 21 days after the first beetle release. Wheat samples from the bins were milled to determine the effects of parasitoid releases on insect fragment counts in flour. In the first year of the study, after 198 days of storage, insect fragment counts were 9.4 and 31 per 50 g flour in the treatment and control bins. However, because of high variability, the means were not significantly different. New grain was used in the second year of the study, and higher numbers of beetles were released. After 131 days of storage, fragment counts averaged 56 and 487/50 g in the treatment and control bins, a reduction in the former of 89%. In the second year of the study, insect myosin in the treatment and control bins averaged 0.27 and 3.23 ng/well, a percentage reduction in the treated bin of 92%. The number of insect damaged kernels (IDK) was significantly lower in the treatment than in the control bins in both years of the study. In the first year, IDK was 6 and 15 IDK/100 g wheat in the treatment and control bins respectively, a reduction in the former of 61%. In the second year, IDK was 12 and 148 IDK/100 g wheat, in the treatment and control bins respectively, a reduction of 92%. This study showed that augmentative releases of parasitoid wasps into bins of stored wheat reduced damage to wheat kernels and the number of insect fragments in flour.

Journal Article↗
PubMed2002

Structure of the N-WASP EVH1 domain-WIP complex: insight into the molecular basis of Wiskott-Aldrich Syndrome.

Missense mutants that cause the immune disorder Wiskott-Aldrich Syndrome (WAS) map primarily to the Enabled/VASP homology 1 (EVH1) domain of the actin regulatory protein WASP. This domain has been implicated in both peptide and phospholipid binding. We show here that the N-WASP EVH1 domain does not bind phosphatidyl inositol-(4,5)-bisphosphate, as previously reported, but does specifically bind a 25 residue motif from the WASP Interacting Protein (WIP). The NMR structure of the complex reveals a novel recognition mechanism-the WIP ligand, which is far longer than canonical EVH1 ligands, wraps around the domain, contacting a narrow but extended surface. This recognition mechanism provides a basis for understanding the effects of mutations that cause WAS.

Amino Acid Sequence↗
PubMed1998

The WASp homologue Las17p functions with the WIP homologue End5p/verprolin and is essential for endocytosis in yeast.

Several end mutations that block the internalisation step of endocytosis in Saccharomyces cerevisiae also affect the cortical actin cytoskeleton [1]. END5 encodes a proline-rich protein (End5p or verprolin) required for a polarised cortical actin cytoskeleton and endocytosis [2,3]. End5p interacts with actin [4], but its exact function is not yet known. To help elucidate End5p function, we sought other End5p-interacting proteins and identified the LAS17/BEE1 gene (encoding the yeast homologue of the human Wiskott-Aldrich Syndrome protein, WASp) as a high-copy-number suppressor of the temperature-sensitive growth and endocytic defects of end5-1 cells (carrying a frameshift mutation affecting the last 213 residues of End5p). LAS17 is unable to suppress a full deletion of END5 (end5 delta), however, suggesting that the defective End5-1p in end5-1 mutants may be stabilised by Las17p. The amino terminus of Las17p interacts with the carboxyl terminus of End5p in the yeast two-hybrid system and similar interactions have been shown between WASp and a mammalian End5p homologue, WASp-interacting protein (WIP) [5]. As las17 delta deletion mutants are blocked in endocytosis, we conclude that Las17p and End5p interact and are essential for endocytosis.

Carrier Proteins↗
PubMed1999

Activation of the yeast Arp2/3 complex by Bee1p, a WASP-family protein.

The Arp2/3 complex is a highly conserved cytoskeletal component that has been implicated in the nucleation of actin filament assembly. Purified Arp2/3 complex has a low intrinsic actin nucleation activity, leading to the hypothesis that an unidentified cellular activator is required for the function of this complex. We showed previously that mutations in the Arp2/3 complex and in Bee1p/Las17p, a member of the Wiskott-Aldrich syndrome protein(WASP) family, lead to a loss of cortical actin structures (patches) in yeast. Bee1p has also been identified as an essential nucleation factor in the reconstitution of actin patches in vitro. Recently, it was reported that WASP-like proteins might interact directly with the Arp2/3 complex through a conserved carboxy-terminal domain. Here, we have shown that Bee1p and the Arp2/3 complex co-immunoprecipitate when expressed at endogenous levels, and that this interaction requires both the Arc15p and Arc19p subunits of the Arp2/3 complex. Furthermore, the carboxy-terminal domain of Bee1p greatly stimulated the nucleation activity of purified Arp2/3 complex in vitro, suggesting a direct role for WASP-family proteins in the activation of the Arp2/3 complex. Interestingly, deletion of the carboxy-terminal domain of Bee1p neither abolished the localization of the Arp2/3 complex, as had been suggested, nor resulted in a severe defect in cortical actin assembly. These results indicate that the function of Bee1p is not mediated entirely through its interaction with the Arp2/3 complex, and that factors redundant with Bee1p might exist to activate the nucleation activity of the Arp2/3 complex.

Actin-Related Protein 2↗
PubMed2001

Listeria protein ActA mimics WASp family proteins: it activates filament barbed end branching by Arp2/3 complex.

Actin-based propulsion of the bacteria Listeria and Shigella mimics the forward movement of the leading edge of motile cells. While Shigella harnesses the eukaryotic protein N-WASp to stimulate actin polymerization and filament branching through Arp2/3 complex, the Listeria surface protein ActA directly activates Arp2/3 complex by an unknown mechanism. Here we show that the N-terminal domain of ActA binds one actin monomer, in a profilin-like fashion, and Arp2/3 complex and mimics the C-terminal domain of WASp family proteins in catalyzing filament barbed end branching by Arp2/3 complex. No evidence is found for side branching of filaments by ActA-activated Arp2/3 complex. Mutations in the conserved acidic (41)DEWEEE(46) and basic (146)KKRRK(150) regions of ActA affect Arp2/3 binding but not G-actin binding. The motility properties of wild-type and mutated Listeria strains in living cells and in the medium reconstituted from pure proteins confirm the conclusions of biochemical experiments. Filament branching is followed by rapid debranching. Debranching is 3-4-fold faster when Arp2/3 is activated by ActA than by the C-terminal domain of N-WASp. VASP is required for efficient propulsion of ActA-coated beads in the reconstituted motility medium, but it does not affect the rates of barbed end branching/debranching by ActA-activated Arp2/3 nor the capping of filaments. VASP therefore affects another still unidentified biochemical reaction that plays an important role in actin-based movement.

Actin-Related Protein 2↗
PubMed2006

Application of an enzyme-linked immunosorbent assay for the analysis of imidacloprid in wiliwili tree, Erythrina sandwicensis O. Deg, for control of the wasp Quadrastichus erythrinae.

A monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for the neonicotinoid insecticide imidacloprid was evaluated for its reproducibility, accuracy, and comparability to results from a conventional high-performance liquid chromatography (HPLC) for the analysis of imidacloprid in the endemic wiliwili tree (Erythrina sandwicensis O. Deg) found in dryland forests and landscapes in Hawaii. Imidacloprid was applied to these wiliwili trees in an attempt to control the newly introduced erythrina gall wasp, Quadrastichus erythrinae Kim. Leaf samples were freeze-dried and extracted with acidic aqueous methanol followed by methylene chloride partitioning. After solvent removal, the extract residue was reconstituted in 1 mL of water/methanol (1:1, v/v) for ELISA; no significant matrix interference was observed at 10-fold or more dilution. The average recoveries of imidacloprid from fortified samples ranged from 78% to 100% by ELISA. The correlation between the ELISA and HPLC results was excellent (r2 = 0.98). Imidacloprid was detected with the ELISA in all treated samples and its level varied in the samples among different treatments and in those from different parts of the trees. The infestation severity rating of leaf samples was inversely related to the concentration of imidacloprid. It is clear that imidacloprid effectively controls the wasps. The ELISA is a suitable method for quantitative and reliable determination of imidacloprid in wiliwili trees and the application provides information to understand how to control the wasps.

Chromatography, High Pressure Liquid↗