Identification of factors responsible for insecticide resistance in Helicoverpa armigera

Moth larvae (Helicoverpa armigera Hübner) collected from field crops were tested for resistance to cypermethrin, fenvalerate, endosulfan, monocrotophos and quinolphos. Larvae were treated with a dose of the pesticide that would kill 99% of the susceptible insects. The percent survival of the resistant strains was determined. Highest seasonal average percentage survival was recorded by fenvalerate (65.0%) followed by cypermethrin (62.4%). Acetylcholinesterase of resistant larvae was less sensitive to monocrotophos and methyl paraoxon. Resistant larvae showed higher activities of esterases, phosphatases and methyl paraoxon hydrolase compared with susceptible larvae. The presence of high activity of esterases was attributed to appearance of extra bands of esterases in native PAGE. The presence of P-glycoprotein expression was detected in resistant larvae using P-gp antibodies; this was not detected in the susceptible larvae. Our results indicate that the high level of resistance detected in the field pests could be because of a combined effect of decreased sensitivity to AChE, higher levels of esterases, phosphatases and the expression of P-gp.     

Three-dimensional molecular-field analyses of octopaminergic agonists for the cockroach neuronal octopamine receptor

The quantitative structure-activity relationship of a set of 40 octopaminergic agonists against receptor 2 in cockroach nervous tissue, was analyzed using molecular-field analysis (MFA). MFA on the study set of those compounds evaluated effectively the energy between a probe and a molecular model at a series of points defined by a rectangular grid. Contour surfaces for the molecular fields were presented and the results provided useful information in the characterization and differentiation of octopaminergic receptor.     

Diet factors responsible for the change of the glucose oxidase activity in labial salivary glands of Helicoverpa armigera

We investigated the change of the glucose oxidase (GOX) activity in labial salivary glands of Helicoverpa armigera larvae fed with the artificial diet or host plant tobacco and the major factors responsible for such a change. Throughout larval development, the labial salivary GOX activities in caterpillars reared on the artificial diet were remarkably higher than those fed with the plant. After fifth-instar plant-fed caterpillars were transferred to the artificial diet, their labial salivary GOX activity increased quickly, which was closely correlated with the time spent feeding on the artificial diet. The total sugar content of the artificial diet was 68 times higher than that of the tobacco leaves. We hypothesized that sugars and secondary metabolites are the possible causes of induction of GOX activity. When fifth-instar caterpillars were fed with tobacco leaves coated with glucose or sucrose, their labial salivary GOX activity was significantly higher than those fed with leaves without sugar coating. Following native PAGE, 1 single band of the labial salivary GOX was observed in all the caterpillars fed with different diets, implying that only the activity of the isoenzyme was changed in response to different diets. Furthermore, the labial salivary GOX activity was determined after caterpillars were fed with artificial diets containing chlorogenic acid, rutin, and quercetin. The results showed that all these phenolic compounds had no effect on the GOX activity. We conclude that sugar in diets was a major factor influencing the labial salivary GOX activity of the larvae. Arch. Insect Biochem. Physiol. 2008.     

Three-dimensional quantitative structure-activity relationship (3D-QSAR) studies of tricyclic oxazolidinones as antibacterial agents

Oxazolidinones exemplified by eprezolid and linezolid are a new class of antibacterials that are active against Gram positive and anaerobic bacteria including methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE) and vancomycin resistant enterococci (VRE). In an effort to have a better antibacterial agent in the oxazolidinone class, we have performed three-dimensional quantitative structure-activity relationship (3D-QSAR) studies for a series of tricyclic oxazolidinones. 3D-QSAR studies were performed using the Comparative Molecular Field Analysis (CoMFA) and Comparative Molecular Similarity Indices Analysis (CoMSIA) procedures. These studies were performed using 42 compounds; the QSAR model was developed using a training set of 33 compounds. The predictive ability of the QSAR model was assessed using a test set of 9 compounds. The predictive 3D-QSAR models have conventional r(2) values of 0.975 and 0.940 for CoMFA and CoMSIA respectively; similarly, cross-validated coefficient q(2) values of 0.523 and 0.557 for CoMFA and CoMSIA, respectively, were obtained. The CoMFA 3D-QSAR model performed better than the CoMSIA model.     

Exclusion cage studies on the impact of predation on Helicoverpa armigera in cotton

The impact of predation on Helicoverpa armigera was studied in four field cage exclusion trials on cotton in Kenya. H. armigera egg cohorts were introduced inside predator‐free and open control cages, and the impact of local predator populations on the cohort was examined. Fourteen days after inoculation, exclusion cages had four times more larvae than controls, indicating a strong impact of predation. Ants and Anthocoridae were the predominant predator groups. Exclusion cages had more damaged fruiting plant parts (squares, flowers and bolls) than the control. In the absence of predators, natural mortality of H. armigera was greater as cotton matured, and is likely to be linked to the host plant condition.     

Ortho effects in quantitative structure-activity relationships for acetylcholinesterase inhibition by aryl carbamates

Ortho-substituted phenyl-N-butyl carbamates (1-9) are characterized as "pseudo-pseudo-substrate" inhibitors of acetylcholinesterase. Since the inhibitors protonate at pH 7.0 buffer solution, the virtual inhibition constants (K'is) of the protonated inhibitors are calculated from the equation, - logK'i = - logKi - logKb. The logarithms of the inhibition constant (Ki), the carbamylation constant (k(c)), and the bimolecular inhibition constant (k(i)) for the enzyme inhibitions by carbamates 1-9 are multiply linearly correlated with the Hammett para-substituent constant (sigma(p)), the Taft-Kutter-Hansch ortho steric constant (E(S)), and the Swan-Lupton ortho polar constant (F). Values of rho, delta, and f for the - logKi-, logk(c)-, and logk(i)-correlations are -0.6, -0.16, 0.7; 0.11, 0.03, -0.3; and - 0.5, - 0.12, 0.4, respectively. The Ki step further divides into two steps: 1) the pre-equilibrium protonation of the inhibitors, Kb step and 2) formation of a negatively charged enzyme-inhibitor Michaelis-Menten complex--virtual inhibition, K'i step. The Ki step has little ortho steric enhancement effect; moreover, the k(c)step is insensitive to the ortho steric effect. The f value of 0.7 for the Ki step indicates that ortho electron-withdrawing substituents of the inhibitors accelerate the inhibition reactions from the ortho polar effect; however, the f value of -0.3 for the k(c)step implies that ortho electron-withdrawing substituents of the inhibitors lessen the inhibition reactions from the ortho polar effect.     

Learning in the nectar foraging behaviour of Helicoverpa armigera

Abstract .1. Learning may enable insects to obtain nectar from flowers more efficiently. Learning in nectar foraging has been shown primarily in studies of bees and butterflies. Here, learning is demonstrated in the nectar foraging behaviour of a noctuid moth, Helicoverpa armigera .
2. The present studies show that: (1) previous experience with a flowering host species increases the probability of that species being selected for nectar foraging, and (2) previous experience of a particular flower type (food source at bottom or top of the corolla tube) increases the likelihood of the food source being found when that flower type is being searched.
3. The implications of these findings for understanding the pattern of oviposition observed in wild populations of this important pest species are discussed.     

Vip3A is responsible for the potency of Bacillus thuringiensis 9816C culture supernatant against Helicoverpa armigera and Spodoptera exigua

Culture supernatant of Bacillus thuringiensis 9816C had high toxicity against Helicoverpa armigera and Spodoptera exigua. However, it lost insecticidal activities after being bathed in boiling water for 5 min. Acrystalliferous mutants of Bt9816C (Bt9816C-NP1 and Bt9816C-NP2) cured of its endogenous plasmids no longer possessed vip3A gene and toxicity. The 89 kD protein which existed in Bt9816C supernatant disappeared in the two mutants' supernatant; nevertheless, the two mutants still exhibited hemolytic and phospholipase C activity as Bt9816C did. The vip3A gene of Bt9816C, vip3Aa18, was cloned and expressed in Escherichia coli BL21. Bioassay demonstrated that the recombinant E. coli had high toxicity against S. exigua. Taken together, it suggested that Vip3A protein was responsible for the toxicity of Bt9816C culture supernatants.     

Evaluation of Helicoverpa armigera Nucleopolyhedrovirus (HearNPV) for Control of Helicoverpa armigera (Lepidoptera: Noctuidae) on Citrus in South Africa

A concentration of 1.15×10⁷ OBs/mL of HearNPV, sprayed using a knapsack, resulted in a 100% reduction in Helicoverpa armigera larval infestation within 7 days on tomato plants in a hot house environment. A 10-fold lower concentration, 1.15×10⁶ OBs/mL, resulted in a 100% reduction within 16 days. These two concentrations reduced damage to tomatoes by 81 and 69%, respectively. In two field trials on navel oranges, the lower concentration and an additional even lower concentration of 7.26×10⁵ OBs/mL, both resulted in a 100% reduction in H. armigera infestation within 14 days or longer. H. armigera damage to fruit was reduced by up to 84 and 75%, respectively, in the two trials. Rejection for export was reduced by 96 and 62%. This could imply a saving of US$339 per hectare compared to the untreated control. These results were better than those achieved with Bacillus thuringiensis and various standard chemical insecticides used in the same trials. Reasons for these results are discussed.     

棉铃虫围食膜蛋白的电泳分离技术

比较了棉铃虫Helicoverpa armigera(Hübner)围食膜不同处理方法,不同围食膜数量,在不同电泳下的分离效果,筛选最佳的围食膜蛋白分离技术。结果表明:冷冻干燥与非冷冻干燥处理围食膜后进行SDS-PAGE电泳分离效果基本相同,建议使用冷冻干燥后处理较好。取1、3、5条围食膜进行SDS-PAGE电泳,都能将围食膜蛋白分离,且分离效果基本一致,建议5条围食膜最合适,具有可靠性和代表性。浓缩胶为5%、分离胶分别为8%、10%和12%时棉铃虫围食膜的SDS-PAGE电泳分离效果差异不大,而分离胶为12%时效果较好。无水三氟利克酸处理围食膜后进行NuPAGE电泳,分离出的围食膜蛋白大约30多种,远远超过上述方法分离的16²0种,且需要的围食膜材料少,分离效果最佳,但费用高。     

Three-dimensional quantitative structure-activity relationship study of the inhibition of Na(+),K(+)-ATPase by cardiotonic steroids using comparative molecular field analysis.

Na(+),K(+)-ATPase is a transmembrane protein that transports sodium and potassium ions across cell membranes during an activity cycle that uses the energy released by ATP hydrolysis. Cardiotonic steroids (digitalis) inhibit this activity and consequently produce a positive inotropic response in the heart. To identify the structural features of the steroids that are important for this inhibition, we have tested the inhibitory properties of 47 cardiotonic and hormonal steroids and developed a three-dimensional quantitative structure-activity relationship (3D-QSAR) model for the inhibition of Na(+),K(+)-ATPase using comparative molecular field analysis (CoMFA). We also developed a 3D-QSAR model for the binding of digoxin to the murine anti-digoxin monoclonal antibody (mAb) 26-10 because we have previously shown that the environment of the binding sites of 26-10 and the enzyme are similar (Kasturi et al. (1998) Biochemistry 37, 6658-6666). These statistically predictive 3D-QSAR models indicate that both binding sites are about 20 A long and have a close fit or complementarity about the beta side of the lactone ring of digitalis. Furthermore, steric bulk about the lactone ring and the alpha sugar may be critical for drug binding. However, the binding site of Na(+),K(+)-ATPase differs from that of mAb in that it has a greater number of electrostatic interactions along the alpha-sugar, steroid, and lactone moieties. In addition, the availability of the structure of the 26-10 Fab-digoxin complex (Jeffrey et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 10310-10314) enabled us to compare the CoMFA-derived contour maps with the known locations for amino acid residues comprising the mAb ligand binding site.     

Synthesis and structure-activity relationship studies of 4,11-diaminonaphtho[2,3-f]indole-5,10-diones

We describe the synthesis of derivatives of 4,11-diaminonaphtho[2,3-f]indole-5,10-dione and their cytotoxicity for human tumor cells that express major determinants of altered anticancer drug response, the efflux pump P-glycoprotein, and non-functional p53. Nucleophilic substitution of methoxy groups in 4,11-dimethoxynaphtho[2,3-f]indole-5,10-dione with various ethylenediamines yielded the derivatives of 4,11-diaminonaphtho[2,3-f]indole-5,10-dione, the indole containing analogues of the antitumor agent ametantrone. The cytotoxicity of novel compounds for multidrug resistant, P-glycoprotein-expressing tumor cells is highly dependent on the N-substituent at the terminal amino group of the ethylenediamine moiety. Whereas p53 null colon carcinoma cells were less sensitive to the reference drug doxorubicin than their counterparts with wild type p53, the majority of novel naphthoindole derivatives were equally potent for both cell lines, regardless of the p53 status.     

Peculiarities [correction of Peculiarites] of the graviperception and graviresponses [correction of graviresponces] in some unicellular eukaryotic organisms

The growth and ultrastructure of Vaucheria sessilis, a siphonaceous alga, were studied during simulated weightlessness using a clinostat and during hypergravity on a centrifuge. Observations, including abnormalities in sporulation and changes in the distribution of nuclei, are presented and discussed.     

Isolation of a cigarette smoke fraction responsible for the inhibition of benzo[a]pyrene metabolism in the isolated perfused rabbit lung

Among the several thousand components of cigarette smoke is a substance or substances capable of inhibiting pulmonary metabolism of nicotine and altering the metabolite profile of procarcinogens such as benzo[a]pyrene (BP). This substance(s) inhibits BP metabolism in the lung in amounts present in a few puffs of cigarette smoke. By a series of extractions and chromatographic methods an active subfraction containing only 1% of the total cigarette smoke condensate (CSC), was isolated. This fraction demonstrated the same inhibition of BP metabolism in the isolate perfused lung (IPL) as the whole smoke. The inhibitor(s) present in this fraction possess amphoteric characteristics. The acidic function is believed to be a phenolic one.     

The mechanism [correction of mehanism] of spatial [correction of spetial] orientation in conditions of G-stress

Six test subjects were subjected to lateral (+Gy) and longitudinal-lateral (+Gz/+Gy) accelerations in a centrifuge with a rotation radius of 6.55 m. During rotation, test subjects were instructed to indicate the position of subjective visual vertical. Results of this study demonstrated that during exposure to +Gy and +Gz/+Gy accelerations, the direction of the indicated subjective vertical approached the direction of the resultant acceleration vector when the lateral component increased. This observed effect decreases with an increase of the longitudinal component of the acceleration. It was suggested that exposure to (i.e. "pulling") high lateral acceleration (up to 2-3 Gy) in highly maneuverable aircraft can hinder spatial orientation of a pilot due to this persistent illusory spatial position as reported above. Our analysis showed that the process of spatial orientation under the conditions of G-load influence becomes more difficult and it is depending on the compromise between visual and vestibular-proprioceptive inputs. On account of this finding, it may be proposed that under conditions of G-load influence, pilots that rely primarily on visual perception may be exposed to higher risk of spatial disorientation.     

Synergy between parasitoids and pathogens for biological control of Helicoverpa armigera in chickpea

The chickpea pod borer, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), is an important pest of many legume crops, including chickpea, Cicer arietinum L. (Fabaceae). We field-tested the efficacy of two pathogens, Helicoverpa armigera nucleopolyhedrosis virus (HaNPV) and Bacillus thuringiensis Berliner subsp. kurstaki (Btk), both alone and in combination with the parasitoid Habrobracon hebetor Say (Hh) (Hymenoptera: Braconidae), for control of H. armigera larvae in chickpea fields. A randomized complete block design was used at two sites, so that wasp release treatments at one site would not contaminate pathogen-only treatments at the other. Pathogens were applied when most pest larvae were in the second instar, and wasp releases were made 2 days later. Number of H. armigera larvae per plant, number of damaged pods, yield (kg ha⁻¹), treatment cost (US$ ha⁻¹), and benefit:cost ratio were all determined. Treatments separated significantly at 14 days after treatment, with the HaNPV+Hh and Btk+Hh treatments providing the best control, suggesting a synergistic impact of the wasp and the pathogens when applied together. These treatments also produced the fewest damaged pods, the highest yields, and the greatest yield increments in comparison to untreated controls. However, costs were also highest for these combined treatments, which reduced their net benefit in comparison to use of Hh or Btk alone, although more efficient mass production of the pathogens might make combined applications feasible. We conclude that both HaNPV and Btk are suitable for integration with releases of H. hebetor against H. armigera in chickpea fields, provided parasitoid releases are delayed until 2 days after pathogen application.     

安徽棉铃虫的生态控制研究

本从气候因子、天敌、耕作栽培措施、诱集技术、生物制剂和转Ｂｔ基因棉等几个方面对安徽棉铃虫种群的生态控制技术研究进行了总结,表明各种生物控制措施对控制棉铃虫的种群是有效的;种群系统分析将是研究棉铃虫生态控制研究的主要方法,建立科学合理的生态控制系统是棉铃虫控制研究的重点。     

The impact of ingested potato type II inhibitors on the production of the major serine proteases in the gut of Helicoverpa armigera

The flowers of the ornamental tobacco produce high levels of a series of 6 kDa serine protease inhibitors (NaPIs) that are effective inhibitors of trypsins and chymotrypsins from lepidopteran species. These inhibitors have a negative impact on the growth and development of lepidopteran larvae and have a potential role in plant protection. Here we investigate the effect of NaPIs on the activity and levels of serine proteases in the gut of Helicoverpa armigera larvae and explore the adaptive mechanisms larvae employ to overcome the negative effects of NaPIs in the diet. Polyclonal antibodies were raised against a Helicoverpa punctigera trypsin that is a target for NaPIs and two H. punctigera chymotrypsins; one that is resistant and one that is susceptible to inhibition by NaPIs. The antibodies were used to optimize procedures for extraction of proteases for immunoblot analysis and to assess the effect of NaPIs on the relative levels of the proteases in the gut and frass. We discovered that consumption of NaPIs did not lead to over-production of trypsins or chymotrypsins but did result in excessive loss of proteases to the frass.     

Kinetic characterization of the group II Helicoverpa armigera nucleopolyhedrovirus propagated in suspension cell cultures: implications for development of a biopesticides production process

Large-scale commercialization of baculovirus biopesticides for the control of insect pests requires a cell culture production process, and knowledge of the infection kinetics is a vital prerequisite for process optimization. Well-characterized kinetic parameters have so far only been reported for the commercially established recombinant Autographa californica nucleopolyhedrovirus (AcMNPV), a Group I NPV. In this work, key infection kinetic parameters of the Group II NPV Helicoverpa armigera nucleopolyhedrovirus (HaSNPV), and its Few Polyhedra (FP) mutant, were well characterized for the first time, in suspension HzAM1 insect cell cultures, to facilitate the scale-up of an HaSNPV-based biopesticide. The FP mutant had a selective advantage over wild-type HaSNPV in cell cultures, and the kinetic analysis showed that this was due to a superior budding rate, rather than a faster binding rate (BR) or longer budding duration. Another finding was that wild-type HaSNPV had very poor infection kinetics when compared with AcMNPV, exhibiting an 18-fold lower BR, a more than 50-fold lower budding rate, and a 60-fold lower extracellular/total progeny virus ratio. Such poor infection kinetics have serious implications during scale-up of an HaSNPV biopesticide production process, including the requirement for large volumes of virus inocula and the difficulty of achieving synchronous infections. Groups I and II NPVs may have very different infection kinetics because of their different envelope fusion proteins. This study is the first to compare the two groups of NPVs in terms of well-characterized cell-specific infection kinetics, and the findings may indicate a phylogenetic basis for kinetic differences.