Effects of genetic and environmental host plant variation on the susceptibility of two noctuids toBacillus thuringiensis

TwoApium graveolens var.rapaceum (L.) cultivars that differ in their suitability for the survival and growth ofSpodoptera exigua (Hübner) andTrichoplusia ni (Hübner) were used to examine the effect of genetic and seasonal environmental variation in host plant suitability on the efficacy ofBacillus thuringiensis subsp.kurstaki (Berliner). The effects of host plant genotype andB. thuringiensis were generally independent, so thatB. thuringiensis efficacy was greatest on the resistant host plant cultivar. Host plant suitability varied within growing season for both insect species but, while host plant suitability decreased with increasing plant age forT. ni, the response ofS. exigua to plant age was not as clear. Within season variation in host plant suitability affectedB. thuringiensis efficacy and the interaction betweenB. thuringiensis and host plant cultivar forS. exigua but not forT. ni. Soluble protein and Folin-Denis phenolic concentrations of host plant tissue were not correlated with changes in host plant suitability to either insect species.     

Effect of the pheromone biosynthesis activating neuropeptide on sex pheromone biosynthesis in Spodoptera littoralis isolated glands

The control of Spodoptera littoralis sex pheromone biosynthesis has been investigated with synthetic pheromone biosynthesis activating neuropeptide (PBAN) and different labeled tracers using an in vitro isolated gland system. Responsiveness of the glands to PBAN stimulation was impaired by careless tissue manipulation. The fact that PBAN is active in the isolated gland system suggests that this might be a target organ for this peptide in S. littoralis. As reported previously with Br-SOG extracts and intact females, label incorporation into the pheromone increased in glands treated with PBAN from all the precursors tested. However, the formation of labeled intermediates from d₅E11–14:Acid also occurred in glands incubated in the absence of the peptide, but the amounts of d₅Z9, E11–14:Acid were lower in PBAN treated glands than in controls. These results indicate that PBAN controls pheromone biosynthesis in S. littoralis by regulating the reduction of acyl moieties. © 1994 Wiley-Liss, Inc.     

Evaluation of monitoring approaches and effects of culture conditions on recombinant protein production in baculovirus-infected insect cells

The baculovirus infection process ofSpodoptera frugiperda (Sf9) insect cells in oxygen-controlled bioreactors in serum-free medium was investigated using a recombinantAutographa californica (AcNPV) virus expressing -galactosidase enzyme as a model system. A variety of monitoring techniques including trypan blue exclusion, fluorescent dye staining, oxygen uptake rate (OUR) measurements, and glucose consumption were applied to infected cells to determine the best way of evaluating cell integrity and assessing the course of baculovirus infection. The metabolism of newly-infected cells increased 90% during the first 24 hours, but as infection proceeded, and cells gradually succumbed to the baculovirus infection, the cytopathic effect of the baculovirus on the cells became evident. Oxygen and glucose uptake rate measurements appeared to more accurately assess the condition of infected cells than conventional trypan blue staining, which tended to overestimate cell viability in the mid stages of infection. The optimal harvest time varied, depending on which technique — SDS-PAGE, chromogenic (ONPG) or fluorometric (C₁₂FDG) — was used to monitor -galactosidase production. Specific -galactosidase production was found to be insensitive to a wide range of culture dissolved oxygen tensions, whereas resuspending cells in fresh medium prior to infection increased volumetric productivity approximately two-fold (800,000 units -galactosidase/ml) compared to cultures infected in batch mode and allowed successful infections to occur at higher cell densities.Abbreviations ONPG ortho-phenyl 2--D-galactopyranoside - OUR oxygen uptake rate (-mol O₂/liter/hour) - q_glucose specific glucose uptake rate (mg glucose/10⁶cell/hour) - q_glutamine specific glutamine uptake rate (mg glutamine/10⁶cell/hour) - qO₂ specific oxygen uptake rate (-mol O₂/10⁶cell/hour) - MOI virus multiplicity of infection (viral plaque forming units/cell)     

Ecology and biology ofNoctuidonema guyanense (Nematoda,aphelenchoididae), an ectoparasite ofSpodoptera frugiperda (Lep., Noctuidae), in French Guiana

Male and female moths ofSpodoptera frugiperda (J. E. Smith), collected in French Guiana between 1980 and 1987 were examined to evaluate the level of parasitism by the Aphelenchoidid ectoparasitic nematodeNoctuidonema guyanense Remillet &; Silvain. The rate of parasitism varies according to the population level ofS. frugiperda, the sex of the moths, number of matings, and age of the females. The results confirm that the transmission of the nematode from one moth to another occurs during mating. The growth of a nematode colony on a moth seems to be limited by the development of the physiological state of the host.     

In Vitro and Molecular Docking Evaluation of Larvicidal Effects of Essential Oils of Five Aromatic Plants on the Fall Armyworm Spodoptera frugiperda JE. Smith (Lepidoptera: Noctuidae) from Ivory Coast

Faced with the serious consequences resulting from the abusive and repeated use of synthetic chemicals, today rethinking crop protection is more than necessary. It is in this context that the essential oils of the Lamiaceae Ocimum gratissimum and Ocimum canum, the Poaceae Cymbopogon citratus and nardus and a Rutaceae Citrus sp. of known chemical compositions were experimented. The evaluation of the larvicidal potential of the essential oils was done by the method of topical application of the test solutions, on the L1−L2 stage larvae from the first generation of S. frugiperda obtained after rearing in an air-conditioned room. Lethal concentrations (LC₁₀, LC₅₀ and LC₉₀) were determined after 48 h. After assessing the larvicidal potential of essential oils, molecular docking was carried out to study protein-ligand interactions and their propensity to bind to insect enzyme sites (AChE). The essential oil of O. gratissimum was the most effective with the lowest lethal concentrations (LC₁₀=0.91 %, LC₅₀=1.91 % and LC₉₀=3.92 %). The least toxic oil to larvae was Citrus sp. (LC₁₀=5.44 %, LC₅₀=20.50 % and LC₉₀=77.41 %). Molecular docking revealed that p-cymene and thymol from O. gratissimum essential oil are structurally similar and bind to the AChE active site via predominantly hydrophobic interactions and a H-bond with Tyr374 in the case of thymol. The essential oil of O. gratissimum constitutes a potential candidate for the development of biological insecticides for the fight against insect pests and for the protection of the environment.     

Larval diet influence on oviposition behaviour inSpodoptera littoralis

Females ofSpodoptera littoralis Boisd. (Lepidoptera: Noctuidae) with different feeding experiences during their larval development were tested for their ovipositional response to methanol extracts of larval frass and semisynthetic diets. The effect of the following frass, diet and diet component extracts was tested: (a) frass fromS. littoralis orAgrotis segetum larvae fed on a potato-based diet; (b) frass fromS. littoralis larvae fed on a wheat germ-based diet; (c) potato and wheat germ-based diets; and (d) potatoes and wheat germ. Ovipositing females without prior experience of the potato diet were deterred by extracts of: (1) larval frass from either species fed on potato diet; (2) the potato-based diet; (3) potato. Also females with experience of the potato diet during only a part of their larval development were deterred from oviposition by frass of larvae reared on the potato diet and by the diet itself. However, for females reared on the potato diet for their entire larval development, oviposition was no longer deterred by either of the three extracts listed above. Extracts of: (1) frass from larvae of either species reared on wheat germ diet: (2) the wheat germ diet; or (3) wheat germ did not significantly affect oviposition. Females with ablated antennae were still deterred by frass extracts from larvae fed on potato diet, when they had been reared on the wheat germ diet. In feeding experiments, larvae of larval stage one and of larval stage three-four reared on either of the two diets preferred to feed on the wheat germ diet. However, the preference was significantly stronger for larvae with no prior contact with the potato diet. The effect of larval experience on the loss of oviposition-deterring activity by extracts of larval frass, diets and diet components is discussed in view of induction and selection.     

Field Evaluation of a Commercial Formulation of the Spodoptera exigua (Lepidoptera: Noctuidae) Nuclear Polyhedrosis Virus for Control of Beet Armyworm on Vegetable Crops in Thailand

Small - scale field trials were conducted in the Kanchanaburi and Ratchaburi provinces of Thailand between December 1994 and March 1996 to gather efficacy data to support a registration dossier for a commercial formulation of a nuclear polyhedrois virus of beet armyworm , Spodoptera exigua, under the tradename Spod - X . A liquid concentrate formula tion of Spod - X was compared with two commercial standards , Bacillus thuringiensis subsp . aizawai (CenTuri 3 . 5 WDG) and the insect growth regulator tebufenozide (Mimic 20F , a benzoylhydrazine ecdysone agonist) , for control of beet armyworm on garden pea , Chinese kale , shallot and table grape . Spod - X applied at 4 - day intervals at 3 . 1 - 12 . 5 1011 occlusion bodies (OBs) ha 1 in a spray volume of 625 - 1000 l ha 1 provided excellent control of beet armyworm on garden pea and grape (75 - 100 % reductions) after the second application . On Chinese kale and shallot , rates of Spod - X in the rate 5 - 60 1011 OBs ha 1 sprayed every 4 days in a spray volume of 1000 l ha 1 provided significant protection from infestation of beet armyworm over the growing season; however , efficacy was poor to moderate . Tank - mixing Spod - X with the pyrethroid , deltamethrin (Decis3EC) , did not improveefficacy on shallot;however , combinations of virus and B. thuringiensis or tebufenozide achieved commercially acceptable levels of crop protection on Chinese kale compared with virus alone . Lower levels of control and crop pro tection observed with all materials on Chinese kale and shallot reflect , in part , the tendency of S. exigua larvae to feed within sheltered sites and directly damage marketable plant parts . In general , all rates of Spod - X performed as well as , or better than , the B. thuringiensis standard (56 . 3 - 105 g ha 1 a.i.) on all crops tested . In general , reductions of beet armyworm larval populations and plant damage provided by all labeled rates of the chemical standard , tebufenozide (125 - 250 g ha 1 a . i .), were marginally greater than all rates of Spod - X tested . On crops such as pea and grape , where beet armyworm predominately feed on exposed leaf surfaces , our data unequivocally support the use of Spod - X for cost - effective , season - long pest management . Strategies to improve the efficacy of virus on shallot and kale are discussed .     

Efficient membrane targeting of the chick nicotinic acetylcholine receptor α-subunit in stable insect cell lines

We have synthesised the -subunit of the chick nicotinic acetylcholine receptor (nAChR) in stable, continuous insect (Spodoptera frugiperda) cell lines. A cDNA was integrated randomly into the insect cell genome under control of a baculovius immediate early gene promoter. Transformed cells were obtained by co-transfection of the insect cells with pIEK1.nAChR, encoding the -subunit cDNA, and pIEK1.neo, encoding the neomycin resistance gene. G-418-resistant clones were selected and expanded into continuous cell lines synthesising the chick nAChR -subunit. Using fluorescence microscopy and ligand binding studies we were able to demonstrate efficient membrane targeting of the receptor subunit in the insect cell plasma membrane. Stable insect cell lines may thus have significant advantages over transient baculovirus vectors for the synthesis and characterisation of heterologous receptor proteins.Abbreviations AcNPV Autographa californica nuclear polyhedrosis virus - BTX -bungarotoxin - BSA bovine serum albumin - FITC Fluoroscein isothiocyanate - G418 geneticin-418 - hpi hours post-infection - ie-1 immediate early 1 gene - nAChR nicotinic acetylcholine receptor alpha subunit - Sf Spodoptera frugiperda - tPA tissue plasminogen activator     

The role of the secondary plant compound 2,5-dihydroxymethyl 3,4-dihydroxypyrrolidine as a feeding inhibitor for insects

The cyclic amino alditol 2,5-dihydroxymethyl 3,4-dihydroxypyrrolidine (DMDP) has recently been shown to be an inhibitor of various glucosidases. We have investigated its role as a plant protective chemical against a range of phytophagous insects by examining its effects on development, feeding behaviour and functioning of taste receptors. We have shown that it can inhibit feeding and can be toxic. The mechanisms by which these effects are achieved are discussed in the context of the molecular structure of the compound.

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Résumé Récemment a été mis en évidence le puissant pouvoir inhibiteur de différents glucosidases. Présenté par l'amino-alditol cyclique, 2,5 dihydroxymethyl 3,4 dihydroxypyrrolidine (DMDP), isolé de graines de la légumineuse Lonchocarpus sericeus. Nous avons examiné le rôle de ce composé secondaire dans la stratégie défensive de la plante contre une gamme d'insectes phytophages, à travers son action sur le développement, le comportement alimentaire et le fonctionnement des récepteurs gustatifs. Nous avons montré que DMDP peut inhiber, suivant la dose, la prise de nourriture et, qu'aprs ingestion (par Spodoptera spp), il peut agir comme une toxine. Par suite de sa structure moléculaire, DMDP peut être considéré, non seulement comme l'analogue d'un sucre, mais aussi comme un alcaloïde. Le bilan des résultats de nos expériences suggère que l'efficacité de DMDP comme phagodissuadant, si elle ne provient pas de sa toxicité, découle principalement de ses propriétés d'alcaloïde.