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Overexpression of poplar wounding-inducible genes in Arabidopsis caused improved resistance against Helicoverpa armigera (Hubner) larvae.

ABSTRACT: Four highly inducible genes of poplar trees, PtdKTI5, PtdWIN4, PtdPOP3 from hybrid poplar (Populus trichocarpa × P. deltoides) and PtKTI2 from trembling aspen (Populus tremuloides Michx.) have been individually transformed into Arabidopsis thaliana for overexpression. High transcriptional level of each transgene in transgenic Arabidopsis lines was confirmed by RT-PCR analysis. The development, body weight and survivorship of cotton bollworm (Helicoverpa armigera) fed on four types of transgenic Arabidopis plants were evaluated in the laboratory. Our data indicated that these four Populus defense-related genes exhibited various degree of insectital activity on larval and postlarval development of cotton bollworm and may be utilized for herbivore resistance improvement in plant genetic engineering.

SUBMITTER: Hu R

PROVIDER: S-EPMC3501947 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature

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Overexpression of poplar wounding-inducible genes in Arabidopsis caused improved resistance against Helicoverpa armigera (Hübner) larvae.

Hu Rongfeng R Wang Jiehua J Ji Yan Y Song Yingjin Y Yang Shaohui S

Breeding science 20120901 3

Four highly inducible genes of poplar trees, PtdKTI5, PtdWIN4, PtdPOP3 from hybrid poplar (Populus trichocarpa × P. deltoides) and PtKTI2 from trembling aspen (Populus tremuloides Michx.) have been individually transformed into Arabidopsis thaliana for overexpression. High transcriptional level of each transgene in transgenic Arabidopsis lines was confirmed by RT-PCR analysis. The development, body weight and survivorship of cotton bollworm (Helicoverpa armigera) fed on four types of transgenic ...[more]

PMID: 23226090

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Cantharidin Impedes Activity of Glutathione S-Transferase in the Midgut of Helicoverpa armigera Hubner.

Project description:Previous investigations have implicated glutathione S-transferases (GSTs) as one of the major reasons for insecticide resistance. Therefore, effectiveness of new candidate compounds depends on their ability to inhibit GSTs to prevent metabolic detoxification by insects. Cantharidin, a terpenoid compound of insect origin, has been developed as a bio-pesticide in China, and proves highly toxic to a wide range of insects, especially lepidopteran. In the present study, we test cantharidin as a model compound for its toxicity, effects on the mRNA transcription of a model Helicoverpa armigera glutathione S-transferase gene (HaGST) and also for its putative inhibitory effect on the catalytic activity of GSTs, both in vivo and in vitro in Helicoverpa armigera, employing molecular and biochemical methods. Bioassay results showed that cantharidin was highly toxic to H. armigera. Real-time qPCR showed down-regulation of the HaGST at the mRNA transcript ranging from 2.5 to 12.5 folds while biochemical assays showed in vivo inhibition of GSTs in midgut and in vitro inhibition of rHaGST. Binding of cantharidin to HaGST was rationalized by homology and molecular docking simulations using a model GST (1PN9) as a template structure. Molecular docking simulations also confirmed accurate docking of the cantharidin molecule to the active site of HaGST impeding its catalytic activity.

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Elevated carboxylesterase activity contributes to the lambda-cyhalothrin insensitivity in quercetin fed Helicoverpa armigera (Hubner).

Project description:Quercetin as one of the key plant secondary metabolite flavonol is ubiquitous in terrestrial plants. In this study, the decrease in sensitivity to lambda-cyhalothrin was observed in quercetin-fed Helicoverpa armigera larvae. In order to figure out the mechanisms underlying the decreased sensitivity of H. armigera larvae to lambda-cyhalothrin by quercetin induction, the changes in carboxylesterase activity and in-vitro hydrolytic metabolic capacity to lambda-cyhalothrin were examined. The LC50 value of quercetin-fed H. armigera larvae to lambda-cyhalothrin showed 2.41-fold higher than that of the control. S, S, S-Tributyl phosphorotrithioate (DEF) treatment showed a synergism effect on lambda-cyhalothrin toxicity to quercetin-fed H. armigera. Moreover, the activity of carboxylesterase was significantly higher in quercetin-fed H. armigera larvae after fed on quercetin for 48 h. The in-vitro hydrolytic metabolic capacity to lambda-cyhalothrin in quercetin-fed H. armigera larvae midgut was 289.82 nmol 3-PBA/mg protein/min, which is significant higher than that in the control group (149.60 nmol 3-PBA/mg protein/min). The elevated CarE enzyme activity and corresponding increased hydrolytic metabolic capacity to lambda-cyhalothrin in quercetin-fed H. armigera contributed to the enhanced tolerance to lambda-cyhalothrin.

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Comparative TMT Proteomic Analysis Unveils Unique Insights into Helicoverpa armigera (Hubner) Resistance in Cajanus scarabaeoides (L.) Thouars.

Project description:Pigeonpea [Cajanus cajan (L.) Millspaugh] is an economically important legume playing a crucial role in the semi-arid tropics. Pigeonpea is susceptible to Helicoverpa armigera (Hübner), which causes devastating yield losses. This pest is developing resistance to many commercially available insecticides. Therefore, crop wild relatives of pigeonpea, are being considered as potential sources of genes to expand the genetic base of cultivated pigeonpea to improve traits such as host plant resistance to pests and pathogens. Quantitative proteomic analysis was conducted using the tandem mass tag platform to identify differentially abundant proteins between IBS 3471 and ICPL 87 tolerant accession and susceptible variety to H. armigera, respectively. Leaf proteome were analysed at the vegetative and flowering/podding growth stages. H. armigera tolerance in IBS 3471 appeared to be related to enhanced defence responses, such as changes in secondary metabolite precursors, antioxidants, and the phenylpropanoid pathway. The development of larvae fed on an artificial diet with IBS 3471 lyophilised leaves showed similar inhibition with those fed on an artificial diet with quercetin concentrations with 32 mg/25 g of artificial diet. DAB staining (3,3'-diaminobenzidine) revealed a rapid accumulation of reactive oxygen species in IBS 3471. We conclude that IBS 3471 is an ideal candidate for improving the genetic base of cultivated pigeonpea, including traits for host plant resistance.

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Gene Cloning, Prokaryotic Expression, and Biochemical Characterization of a Soluble Trehalase in Helicoverpa armigera Hubner (Lepidoptera: Noctuidae).

Project description:Trehalase is an indispensable component of insect hemolymph that plays important role in energy metabolism and stress resistance. In this study, we cloned and expressed the gene encoding soluble trehalase (HaTreh-1) of Helicoverpa armigera (cotton bollworm) and characterized the enzyme. HaTreh-1 had a full-length open reading frame encoding a protein of 571 amino acids. Sequence comparison indicated that HaTreh-1 was similar to some known insect trehalases. Two essential active sites (D321 and E519) and three essential residues (R168, R221, and R286) were conserved in HaTreh-1. The recombinant trehalase was expressed in Escherichia coli and purified by nickel exchange chromatography. Molecular weight of the recombinant protein was about 71 kDa, and the optimum HaTreh-1 enzyme activity is at 55°C with pH 6.0. Enzymatic assays showed a Km value of 72.8 mmol/liter and a Vmax value of 0.608 mmol/(liter·min). Inhibition assays in vitro indicated that castanospermine, a polyhydroxylated alkaloid, was an effective competitive inhibitor of trehalase with a Ki value of 6.7 μmol/liter. The inhibitor action of castanospermine was linked to its modification effect on trehalase structure. The circular dichroism spectrum showed that the percentage of α-helix increased under the presence of castanospermine. Results of our study will aid in developing effective trehalase inhibitors for controlling H. armigera in the future.

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Molecular Characterization and Function Analysis of the Vitellogenin Receptor from the Cotton Bollworm, Helicoverpa armigera (Hubner) (Lepidoptera, Noctuidae).

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Cannibalism Affects Core Metabolic Processes in Helicoverpa armigera Larvae-A 2D NMR Metabolomics Study.

Project description:Cannibalism is known in many insect species, yet its impact on insect metabolism has not been investigated in detail. This study assessed the effects of cannibalism on the metabolism of fourth-instar larvae of the non-predatory insect Helicoverpa armigera (Lepidotera: Noctuidea). Two groups of larvae were analyzed: one group fed with fourth-instar larvae of H. armigera (cannibal), the other group fed with an artificial plant diet. Water-soluble small organic compounds present in the larvae were analyzed using two-dimensional nuclear magnetic resonance (NMR) and principal component analysis (PCA). Cannibalism negatively affected larval growth. PCA of NMR spectra showed that the metabolic profiles of cannibal and herbivore larvae were statistically different with monomeric sugars, fatty acid- and amino acid-related metabolites as the most variable compounds. Quantitation of ¹H-(13)C HSQC (Heteronuclear Single Quantum Coherence) signals revealed that the concentrations of glucose, glucono-1,5-lactone, glycerol phosphate, glutamine, glycine, leucine, isoleucine, lysine, ornithine, proline, threonine and valine were higher in the herbivore larvae.

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Sub-lethal doses of Nucleopolyhedrosis Virus and synthetic ınsecticides alter the biological parameters of Helicoverpa armigera Hubner (Lepidoptera: Noctuidae).

Project description:Resistance management is very important for devising control strategies of polyphagous insect-pests like Helicoverpa armigera Hübner (Lepidoptera: Noctuidae). Considering the importance of resistance management, demographic features of selected and unselected populations of H. armigera were studied in 6 different treatments viz. emamectin benzoate, Helicoverpa armigera Nucleopolyhedrosis Virus (HaNPV), emamectin benzoate+HaNPV, spinetoram, spinetoram+HaNPV and control. Higher values for fecundity, intrinsic rate, the finite rate of increase (λ) were recorded in the control of selected as compared to the rest of treatment. Similarly, higher values for these population parameters viz. oviposition days, fecundity, intrinsic rate, the finite rate of increase were calculated in the unselected control. Similarly, net reproductive rate (R0) for selected and unselected control was higher as compared to the rest of the treatments. It may happen because these kinds of selection pressures can result in decreased fitness of the test insect thus decreased fitness of H. armigera in different treatments was observed as compared to the control. Additionally, quicker development of susceptible insects was observed because susceptible insects were growing without any stressor (xenobiotics) as compared to the rest which contributed to their faster development.

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Effect of Different Light Spectrum in Helicoverpa armigera Larvae during HearNPV Induced Tree-Top Disease.

Project description:Lepidopteran larvae upon infection by baculovirus show positive photo-tactic movement during tree-top disease. In light of many insects exploiting specific spectral information for the different behavioral decision, each spectral wavelength of light is an individual parsimonious candidate for such behavior stimulation. Here, we investigated the responses of third instar Helicoverpa armigera larvae infected by Helicoverpa armigera nucleopolyhedrovirus (HearNPV) to white (broad-spectrum), blue (450⁻490 nm), UVA (320⁻400 nm), and UVB (290⁻320 nm) lights for the tree-top disease. Our findings suggest that tree-top phenomenon is induced only when the light is applied from above. Blue, white and UVA lights from above induced tree-top disease, causing infected larvae to die in an elevated position compared to those larvae living in the complete dark. In contrast, UVB from above did not induce tree-top disease. Blue light exerted the maximum photo-tactic response, significantly (p < 0.01) higher than white light. The magnitude of the response decreased with decreasing wavelength to UVA, and no response at UVB. Our results suggested that the spectral wavelength of the light has a significant effect on the induction of the tree-top disease in H. armigera third instar larvae infected with HearNPV.

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Susceptibility Profiles of Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) to Deltamethrin Reveal a Contrast between the Northern and the Southern Benin.

Project description:Helicoverpa armigera is an indigenous species in Africa and has been reported in the destruction of several crops in Benin. Management of H. armigera pest is mainly focused on the use of synthetic pyrethroids, which may contribute to resistance selection. This study aimed to screen the susceptibility pattern of field populations of H. armigera to deltamethrin in Benin. Relevant information on the type of pesticides used by farmers were gathered through surveys. Collected samples of Helicoverpa (F0) were reared to F1. F0 were subjected to morphological speciation followed by a confirmation using restriction fragment length polymorphism coupled with a polymerase chain reaction (RFLP-PCR). F1 (larvae) were used for insecticide susceptibility with deltamethrin alone and in the presence of the P450 inhibitor Piperonyl Butoxide (PBO). Deltamethrin and lambda-cyhalothrin were the most used pyrethroids in tomato and cotton farms respectively. All field-sampled Helicoverpa were found to be H. armigera. Susceptibility assays of H. armigera to deltamethrin revealed a high resistance pattern in cowpea (resistance factor (RF) = 2340), cotton (RF varying from 12 to 516) and tomato (RF=85) farms which is a concern for the control of this major polyphagous agricultural pest. There was a significant increase of mortality when deltamethrin insecticide was combined with piperonyl butoxide (PBO), suggesting the possible involvement of detoxification enzymes such as oxidase. This study highlights the presence of P450 induced metabolic resistance in H. armigera populations from diverse cropping systems in Benin. The recorded high levels of deltamethrin resistance in H. armigera is a concern for the control of this major agricultural pest in Benin as the country is currently embarking into economical expansion of cotton, vegetables and grain-legumes cropping systems.

| S-EPMC6603559 | biostudies-literature