In Silico and in Vivo Investigations of Bioagent Helicoverpa Nucleopolyhedrovirus against Helicoverpa armigera in Chickpea

2021 ◽  
Vol 9 (2) ◽  
pp. 21-32
Author(s):  
Ritu Srivastava ◽  
◽  
Amritesh Chandra Shukla ◽  
Keyword(s):  
Green Tea ◽  
Rice Bran ◽  
Helicoverpa Armigera ◽  
Maximum Yield ◽  
3D Structure ◽  
Homology Model ◽  
In Vitro Toxicity ◽  
Helicoverpa Armígera

During investigations; homology model of 3D-structure was built for sequence of polyhedrin protein of Helicoverpa armigera nucleopolyhedrovirus, containing 246 amino acids (Accession: ACI05106.1 GI: 205946055), and evaluated through multiple tools/ applications to judge extent of accuracy in light of existing crystal structure. Further, in vivo experiments were conducted and determined response of different adjuvants with HaNPV and their efficacy. The pooled mean mortality of larvae exposed to virus mixed with 5% green tea and 5% rice bran filtrates (8.3 larvae per 25 plants) was differ significantly from control (15.8 larvae per 25 plants), suggesting that UV protectants & diet enhancer (mannitol) has ability to protect stability of virulence of the virus, under field conditions. The minimum percent pod damage of 8.6% and maximum yield of 1604.8 Kg ha-1 at harvesting was recorded with formulation of indigenous BHA virus isolate @ 2.2 x 105 POBs mL-1 mixed with Roket @50 ppm; followed by formulation with mannitol (@ 1% + green tea 5% + 5% rice bran filtrates) with percent pod damage of 16.8 % and yield of 1045.8 Kg ha-1 of chickpea. Furthermore, in vitro toxicity of fresh virus suspension @ 250 mL ha-1 was recorded more toxic in terms of percent mortality and LT50 (5.65 days). However, three months stored HaNPV formulations [(A) mannitol @ 1%+ green tea@ 5% and (B) mannitol @ 1% + green tea 5% + 5% rice bran filtrates] were more effective in larval reduction with LT50 of 7.89 and 6.00 days, respectively. Virus mixed with 5% green tea and 5% rice bran filtrates gave stability to formulation up-to one year with LT50 of 7.64 days. Findings showed that HaNPV formulations with mannitol (B) have potential that can be used in integrated manner with other IPM practices, to reduce the use of toxic synthetic pesticides in chickpea.

scholarly journals Potensi Nematoda Patogen Serangga Steinernema spp. dalam Pengendalian Hama Utama Tanaman Kapas

2016 ◽  
Vol 1 (2) ◽  
pp. 101
Author(s):  
Heri Prabowo ◽  
I.G.A.A. Indrayani
Keyword(s):  
Helicoverpa Armigera ◽  
Insect Pests ◽  
Soil Surface ◽  
Tenebrio Molitor ◽  
Pectinophora Gossypiella ◽  
Helicoverpa Armígera ◽  
Soy Bean Oil

<p>Steinernema spp. memiliki potensi untuk mengendalikan hama tanaman kapas seperti Helicoverpa armigera dan Pectinophora gossypiella. Steinernema spp. mampu menyebabkan mortalitas P. gossypiella dan H. armi-gera berturut-turut sebesar 31,6–55,4 dan 46,3–63,8%. Steinernema spp. memiliki kemampuan membunuh lebih baik pada P. gossypiella, sedangkan kemampuan reproduksi dalam inangnya lebih baik pada H. armi-gera. Steinernema spp. mampu menginfeksi serangga inang lebih baik pada stadium ulat lebih tua diban-dingkan stadium muda. Steinernema spp. dapat diproduksi secara in vivo dan in vitro. Produksi secara in vivo dapat menggunakan Tenebrio molitor, Tirathaba rufivena, dan Attacus atlas. Produksi secara in vitro dapat menggunakan usus ayam, lemak sapi, dan minyak kedelai. Perlu dikembangkan formulasi Steinerne-ma spp. yang murah dan efektif untuk mengendalikan hama di atas permukaan tanah. Selain itu diperlukan pencarian isolat Steinernema spp. yang virulen dan cepat membunuh hama sasaran.</p><p> </p><p>Steinernema spp. could be potentially used for controlling H. armigera and P. gossypiella on cotton. Steiner-nema spp. causes mortality on P. gossypiella and H. armigera 31,6–55,4 and 46,3–63,8% respectively. The nematode causes a higher mortality on P. gossypiella than on H. armigera, however, produces more juvenile infective on H. armigera than on P. gossypiella. Higher successful infections of Steinernema spp. occurs on late larval stadium than on early one. Production of Steinernema spp. can be in vivo using Tenebrio molitor, Tirathaba rufivena, and Attacus atlas; and in vitro using chicken intestinum, cow lipid, and soy bean oil. For effecttively use, this nematode need to be formulated especially for controlling insect pests on soil surface, as well as finding the more virulent isolates against the target insects.</p>

Effects of dietary quercetin on performance and cytochrome P450 expression of the cotton bollworm, Helicoverpa armigera

2015 ◽  
Vol 105 (6) ◽  
pp. 771-777 ◽  
Author(s):  
D. Liu ◽  
Y. Yuan ◽  
M. Li ◽  
X. Qiu
Keyword(s):  
Cytochrome P450 ◽  
Helicoverpa Armigera ◽  
Adult Emergence ◽  
Cotton Bollworm ◽  
Molecular Response ◽  
Terrestrial Plants ◽  
P450 Expression ◽  
Helicoverpa Armígera

AbstractQuercetin is ubiquitous in terrestrial plants. The cotton bollworm Helicoverpa armigera as a highly polyphagous insect has caused severe crop losses. Until now, interactions between this pest and quercetin are poorly understood at the biochemical and molecular levels. In this study, we investigated the in vivo effects of quercetin on performance of cotton bollworm and on cytochrome P450 (P450) expression. Deleterious effects of quercetin on the performance of the cotton bollworm, including growth, survival, pupation and adult emergence were observed after oral administration of 3 and 10 mg g−1 quercetin to larvae since the third instar, whereas no significant toxic effect was found at 0.1 mg g−1 quercetin treatment. Piperonyl butoxide treatment enhanced the toxicity of quercetin. In vitro metabolism studies showed that quercetin was rapidly transformed by gut enzymes of fifth instar larvae of the cotton bollworm. qRT–PCR results revealed that the effect of quercetin on P450 expression was tissue- and dose-specific. Quercetin regulated P450 expression in a mild manner, and it could serve as P450 inducer (CYP337B1, CYP6B6) or repressor (CYP337B1, CYP6B7, CYP6B27, CYP9A14, CYP6AE11, and CYP4M7). These findings are important for advancing our understanding of the biochemical and molecular response of insects to plant toxins and have implications for a smart pest control.

scholarly journals New Resistance Mechanism in Helicoverpa armigera Threatens Transgenic Crops Expressing Bacillus thuringiensis Cry1Ac Toxin

2005 ◽  
Vol 71 (5) ◽  
pp. 2558-2563 ◽  
Author(s):  
Robin V. Gunning ◽  
Ho T. Dang ◽  
Fred C. Kemp ◽  
Ian C. Nicholson ◽  
Graham D. Moores
Keyword(s):  
Bacillus Thuringiensis ◽  
Helicoverpa Armigera ◽  
Resistance Mechanism ◽  
Transgenic Cotton ◽  
In Vivo Studies ◽  
Cry1ac Toxin ◽  
Helicoverpa Armígera ◽  
History Of

ABSTRACT In Australia, the cotton bollworm, Helicoverpa armigera, has a long history of resistance to conventional insecticides. Transgenic cotton (expressing the Bacillus thuringiensis toxin Cry1Ac) has been grown for H. armigera control since 1996. It is demonstrated here that a population of Australian H. armigera has developed resistance to Cry1Ac toxin (275-fold). Some 70% of resistant H. armigera larvae were able to survive on Cry1Ac transgenic cotton (Ingard) The resistance phenotype is inherited as an autosomal semidominant trait. Resistance was associated with elevated esterase levels, which cosegregated with resistance. In vitro studies employing surface plasmon resonance technology and other biochemical techniques demonstrated that resistant strain esterase could bind to Cry1Ac protoxin and activated toxin. In vivo studies showed that Cry1Ac-resistant larvae fed Cy1Ac transgenic cotton or Cry1Ac-treated artificial diet had lower esterase activity than non-Cry1Ac-fed larvae. A resistance mechanism in which esterase sequesters Cry1Ac is proposed.

scholarly journals Helicoverpa armigera nucleopolyhedrovirus occlusion-derived virus-associated protein, HA100, affects oral infectivity in vivo but not virus replication in vitro

2011 ◽  
Vol 92 (6) ◽  
pp. 1324-1331 ◽  
Author(s):  
Sijiani Luo ◽  
Yanfang Zhang ◽  
Xushi Xu ◽  
Marcel Westenberg ◽  
Just M. Vlak ◽  
...  
Keyword(s):  
Virus Replication ◽  
Helicoverpa Armigera ◽  
Virus Production ◽  
Control Virus ◽  
Helicoverpa Armígera ◽  
Occlusion Bodies ◽  
Lethal Time ◽  
Budded Virus

ORF100 (ha100) of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) has been reported as one of the unique genes of group II alphabaculoviruses encoding a protein located in the occlusion-derived virus (ODV) envelope and nucleocapsid. The protein consists of 510 aa with a predicted mass of 58.1 kDa and is a homologue of poly(ADP–ribose) glycohydrolase in eukaryotes. Western blot analysis detected a 60 kDa band in HearNPV-infected HzAM1 cells starting at 18 h post-infection. Transient expression of GFP-fused HA100 in HzAM1 cells resulted in cytoplasmic localization of the protein, but after superinfection with HearNPV, GFP-fused HA100 was localized in the nucleus. To study the function of HA100 further, an ha100-null virus was constructed using bacmid technology. Viral one-step growth curve analyses showed that the ha100-null virus had similar budded virus production kinetics to that of the parental virus. Electron microscopy revealed that deletion of HA100 did not alter the morphology of ODVs or occlusion bodies (OBs). However, bioassays in larvae showed that the 50 % lethal concentration (LC50) value of HA100-null OBs was significantly higher than that of parental OBs; the median lethal time (LT50) of ha100-null OBs was about 24 h later than control virus. These results indicate that HA100 is not essential for virus replication in vitro. However, it significantly affects the oral infectivity of OBs in host insects, suggesting that the association HA100 with the ODV contributes to the infectivity of OBs in vivo.

scholarly journals Incorporation of GP64 into Helicoverpa armigera nucleopolyhedrovirus enhances virus infectivity in vivo and in vitro

2012 ◽  
Vol 93 (12) ◽  
pp. 2705-2711 ◽  
Author(s):  
Shu Shen ◽  
Yinyin Gan ◽  
Manli Wang ◽  
Zhihong Hu ◽  
Hualin Wang ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Helicoverpa Armigera ◽  
Fusion Proteins ◽  
Virus Infectivity ◽  
F Protein ◽  
Recombinant Viruses ◽  
Group I ◽  
Helicoverpa Armígera

The envelope fusion proteins of baculoviruses, glycoprotein GP64 from group I nucleopolyhedrovirus (NPV) or the F protein from group II NPV and granulovirus, are essential for baculovirus morphogenesis and infectivity. The F protein is considered the ancestral baculovirus envelope fusion protein, while GP64 is a more recent evolutionary introduction into baculoviruses and exhibits higher fusogenic activity than the F protein. Each of the fusion proteins is required by the respective virus to spread infection within larval tissues. A recombinant Helicoverpa armigera NPV (HearNPV) expressing GP64 from Autographa californica multiple nucleopolyhedrovirus, vHaBac-gp64-egfp, was constructed, which still retained the native F protein, and its infectivity was assayed in vivo and in vitro. Analyses by one-step growth curve to determine viral titre and by quantitative PCR to determine viral DNA copy number showed that vHaBac-gp64-egfp was more infectious in vitro than the control, vHaBac-egfp. The polyhedrin gene (polh) was reintroduced into the recombinant viruses and bioassays showed that vHaBac-gp64-polh accelerated the mortality of infected larvae compared with the vHaBac-egfp-polh control, and the LC50 (median lethal concentration) of vHaBac-gp64-polh was reduced to approximately 20 % of that of vHaBac-egfp-polh. Therefore, incorporation of GP64 into HearNPV budded virions improved virus infectivity both in vivo and in vitro. The construction of this bivalent virus with a more efficient fusion protein could improve the use of baculoviruses in different areas such as gene therapy and biocontrol.

scholarly journals Bacillus thuringiensis Bel Protein Enhances the Toxicity of Cry1Ac Protein to Helicoverpa armigera Larvae by Degrading Insect Intestinal Mucin

2009 ◽  
Vol 75 (16) ◽  
pp. 5237-5243 ◽  
Author(s):  
Shangling Fang ◽  
Li Wang ◽  
Wei Guo ◽  
Xia Zhang ◽  
Donghai Peng ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Helicoverpa Armigera ◽  
Trichoplusia Ni ◽  
Knockout Mutant ◽  
Gene Encoding ◽  
In Vitro Degradation ◽  
Intestinal Mucin ◽  
Helicoverpa Armígera ◽  
Cry1ac Protein

ABSTRACT Bacillus thuringiensis has been used as a bioinsecticide to control agricultural insects. Bacillus cereus group genomes were found to have a Bacillus enhancin-like (bel) gene, encoding a peptide with 20 to 30% identity to viral enhancin protein, which can enhance viral infection by degradation of the peritrophic matrix (PM) of the insect midgut. In this study, the bel gene was found to have an activity similar to that of the viral enhancin gene. A bel knockout mutant was constructed by using a plasmid-free B. thuringiensis derivative, BMB171. The 50% lethal concentrations of this mutant plus the cry1Ac insecticidal protein gene were about 5.8-fold higher than those of the BMB171 strain. When purified Bel was mixed with the Cry1Ac protein and fed to Helicoverpa armigera larvae, 3 μg/ml Cry1Ac alone induced 34.2% mortality. Meanwhile, the mortality rate rose to 74.4% when the same amount of Cry1Ac was mixed with 0.8 μg/ml of Bel. Microscopic observation showed a significant disruption detected on the midgut PM of H. armigera larvae after they were fed Bel. In vitro degradation assays showed that Bel digested the intestinal mucin (IIM) of Trichoplusia ni and H. armigera larvae to various degrading products, similar to findings for viral enhancin. These results imply Bel toxicity enhancement depends on the destruction of midgut PM and IIM, similar to the case with viral enhancin. This discovery showed that Bel has the potential to enhance insecticidal activity of B. thuringiensis-based biopesticides and transgenic crops.

scholarly journals In Vivo and In Vitro Toxicity Evaluation of Polyprenols Extracted from Ginkgo biloba L. Leaves

Molecules ◽  
2015 ◽  
Vol 20 (12) ◽  
pp. 22257-22271 ◽  
Author(s):  
Cheng-Zhang Wang ◽  
Jiao-Jiao Yuan ◽  
Wen-Jun Li ◽  
Hong-Yu Zhang ◽  
Jian-Zhong Ye
Keyword(s):  
Ginkgo Biloba ◽  
In Vitro Toxicity ◽  
Toxicity Evaluation

Integration of in vitro neurotoxicity data with biokinetic modelling for the estimation of in vivo neurotoxicity

2007 ◽  
Vol 26 (4) ◽  
pp. 333-338 ◽  
Author(s):  
Anna Forsby ◽  
Bas Blaauboer
Keyword(s):  
Exposure Period ◽  
Cellular Protein ◽  
In Vitro Toxicity ◽  
Target Tissue ◽  
Receptor Function ◽  
Human Neuroblastoma ◽  
Protein Levels ◽  
Effective Doses

Risk assessment of neurotoxicity is mainly based on in vivo exposure, followed by tests on behaviour, physiology and pathology. In this study, an attempt to estimate lowest observed neurotoxic doses after single or repeated dose exposure was performed. Differentiated human neuroblastoma SH-SY5Y cells were exposed to acrylamide, lindane, parathion, paraoxon, phenytoin, diazepam or caffeine for 72 hours. The effects on protein synthesis and intracellular free Ca2+concentration were studied as physiological endpoints. Voltage operated Ca2 +channel function, acetylcholine receptor function and neurite degenerative effects were investigated as neurospecific endpoints for excitability, cholinergic signal transduction and axonopathy, respectively. The general cytotoxicity, determined as the total cellular protein levels after the 72 hours exposure period, was used for comparison to the specific endpoints and for estimation of acute lethality. The lowest concentration that induced 20% effect (EC 20) obtained for each compound, was used as a surrogate for the lowest neurotoxic level (LOEL) at the target site in vivo. The LOELs were integrated with data on adsorption, distribution, metabolism and excretion of the compounds in physiologically-based biokinetic (PBBK) models of the rat and the lowest observed effective doses (LOEDs) were estimated for the test compounds. A good correlation was observed between the estimated LOEDs and experimental LOEDs found in literature for rat for all test compounds, except for diazepam. However, when using in vitro data from the literature on diazepam's effect on gamma-amino butyric acid (GABA)A receptor function for the estimation of LOED, the correlation between the estimated and experimental LOEDs was improved from a 10 000-fold to a 10-fold difference. Our results indicate that it is possible to estimate LOEDs by integrating in vitro toxicity data as surrogates for lowest observed target tissue levels with PBBK models, provided that some knowledge about toxic mechanisms is known. Human & Experimental Toxicology (2007) 26, 333—338

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