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Sample records for helicoverpa armigera nucleopolyhedrovirus

  1. Genomic Sequences of Five Helicoverpa armigera Nucleopolyhedrovirus Genotypes from Spain That Differ in Their Insecticidal Properties

    PubMed Central

    Arrizubieta, Maite; Williams, Trevor; Caballero, Primitivo

    2015-01-01

    Helicoverpa armigera nucleopolyhedrovirus (HearNPV) has proved effective as the basis for various biological insecticides. Complete genome sequences of five Spanish HearNPV genotypes differed principally in the homologous regions (hrs) and the baculovirus repeat open reading frame (bro) genes, suggesting that they may be involved in the phenotypic differences observed among genotypes. PMID:26067949

  2. Genomic Sequences of Five Helicoverpa armigera Nucleopolyhedrovirus Genotypes from Spain That Differ in Their Insecticidal Properties.

    PubMed

    Arrizubieta, Maite; Simón, Oihane; Williams, Trevor; Caballero, Primitivo

    2015-06-11

    Helicoverpa armigera nucleopolyhedrovirus (HearNPV) has proved effective as the basis for various biological insecticides. Complete genome sequences of five Spanish HearNPV genotypes differed principally in the homologous regions (hrs) and the baculovirus repeat open reading frame (bro) genes, suggesting that they may be involved in the phenotypic differences observed among genotypes. Copyright © 2015 Arrizubieta et al.

  3. Selection of a nucleopolyhedrovirus isolate from Helicoverpa armigera as the basis for a biological insecticide.

    PubMed

    Arrizubieta, Maite; Williams, Trevor; Caballero, Primitivo; Simón, Oihane

    2014-06-01

    The cotton bollworm, Helicoverpa armigera, is an insect that causes damage in a wide range of crops in Spain. Seven isolates of H. armigera single nucleopolyhedrovirus (HearSNPV) from the Iberian Peninsula were subjected to molecular and biological characterization and compared with a Chinese genotype (HearSNPV-G4). The estimated sizes of the Iberian genomes varied between 116.2 and 132.4 kb, compared to 131.4 kb of the HearSNPV-G4 reference genome. Phylogenetic analysis based on the lef-8, lef-9 and polh genes revealed that the Iberian strains were more closely related to one another than to other HearSNPV isolates. Occlusion body (OB) concentration-mortality responses (LC50 values) did not differ significantly among Iberian isolates when tested against a Helicoverpa armigera colony from Oxford (UK). Despite being the fastest killing isolate, HearSNPV-SP1 was as productive as isolates with lower virulence, with an average yield of 3.1 × 10(9) OBs larva(-1) . OBs of HearSNPV-SP1 and HearSNPV-G4 were similarly pathogenic against a recently established colony from southern Spain, although HearSNPV-SP1 was faster killing than HearSNPV-G4 against a range of instars. The insecticidal properties of HearSNPV-SP1 mean that this strain is likely to prove useful as the basis for a biological insecticide for control of Helicoverpa armigera in Spain. © 2013 Society of Chemical Industry.

  4. Characterization of a new Helicoverpa armigera nucleopolyhedrovirus variant causing epizootic on a previously unreported host, Helicoverpa gelotopoeon (Lepidoptera: Noctuidae).

    PubMed

    Ferrelli, M L; Taibo, C; Fichetti, P; Sciocco-Cap, A; Arneodo, J D

    2016-07-01

    This paper reports the first biological and molecular characterization of a nucleopolyhedrovirus isolated from the soybean and cotton pest Helicoverpa gelotopoeon. Studies were performed following a virus outbreak in a rearing facility and in wild H. gelotopoeon populations in Córdoba, Argentina. Host identity was corroborated by partial sequencing of the COI gene. Scanning electron microscope observations of purified OBs revealed their polyhedral morphology and an average diameter of 0.89±0.14μm. Ultrathin sections of infected larvae examined by transmission electron microscopy showed the intranuclear occurrence of polyhedra and virus particles in fat body cells. Nucleocapsids were singly enveloped. Phylogenetic analysis of lef-8, lef-9, polh, orf5/5b and hr3-orf62 viral sequences identified this new NPV isolate (hereafter HegeSNPV) as a variant of Helicoverpa armigera nucleopolyhedrovirus (HearNPV). Furthermore, HegeSNPV was closely related to the so-called "HzSNPV Group" within HearNPV, although having particular characteristics.

  5. Horizontal and vertical transmission of wild-type and recombinant Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus.

    PubMed

    Zhou, Mingzhe; Sun, Xiulian; Sun, Xincheng; Vlak, Just M; Hu, Zhihong; van der Werf, Wopke

    2005-06-01

    Transmission plays a central role in the ecology of baculoviruses and the population dynamics of their hosts. Here, we report on the horizontal and vertical transmission dynamics of wild-type Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HaSNPV-WT) and a genetically modified variant (HaSNPV-AaIT) with enhanced speed of action through the expression of an insect-selective scorpion toxin (AaIT). In caged field plots, horizontal transmission of both HaSNPV variants was greatest when inoculated 3rd instar larvae were used as infectors, transmission was intermediate with 2nd instar infectors and lowest with 1st instar infectors. Transmission was greater at a higher density of infectors (1 per plant) than at a lower density (1 per 4 plants); however, the transmission coefficient (number of new infections per initial infector) was lower at the higher density of infectors than at the lower density. HaSNPV-AaIT exhibited a significantly lower rate of transmission than HaSNPV-WT in the field cages. This was also the case in open field experiments. In the laboratory, the vertical transmission of HaSNPV-AaIT from infected females to offspring of 16.7+/-2.1% was significantly lower than that of HaSNPV-WT (30.9+/-2.9%). Likewise, in the field, vertical transmission of HaSNPV-AaIT (8.4+/-1.1%) was significantly lower than that of HaSNPV-WT (12.6+/-2.0%). The results indicate that the recombinant virus will be transmitted at lower rates in H. armigera populations than the wild-type virus. This may potentially affect negatively its long-term efficacy as compared to wild-type virus, but contributing positively to its biosafety.

  6. Identification of protein-protein interactions of the occlusion-derived virus-associated proteins of Helicoverpa armigera nucleopolyhedrovirus.

    PubMed

    Peng, Ke; Wu, Minzhi; Deng, Fei; Song, Jingjiao; Dong, Chunsheng; Wang, Hualin; Hu, Zhihong

    2010-03-01

    The purpose of this study was to identify protein-protein interactions among the components of the occlusion-derived virus (ODV) of Helicoverpa armigera nucleopolyhedrovirus (HearNPV), a group II alphabaculovirus in the family Baculoviridae. To achieve this, 39 selected genes of potential ODV structural proteins were cloned and expressed in the Gal4 yeast two-hybrid (Y2H) system. The direct-cross Y2H assays identified 22 interactions comprising 13 binary interactions [HA9-ODV-EC43, ODV-E56-38K, ODV-E56-PIF3, LEF3-helicase, LEF3-alkaline nuclease (AN), GP41-38K, GP41-HA90, 38K-PIF3, 38K-PIF2, VP80-HA100, ODV-E66-PIF3, ODV-E66-PIF2 and PIF3-PIF2] and nine self-associations (IE1, HA44, LEF3, HA66, GP41, CG30, 38K, PIF3 and P24). Five of these interactions - LEF3-helicase and LEF3-AN, and the self-associations of IE1, LEF3 and 38K - have been reported previously in Autographa californica multiple nucleopolyhedrovirus. As HA44 and HA100 were two newly identified ODV proteins of group II viruses, their interactions were further confirmed. The self-association of HA44 was verified with a His pull-down assay and the interaction of VP80-HA100 was confirmed by a co-immunoprecipitation assay. A summary of the protein-protein interactions of baculoviruses reported so far, comprising 68 interactions with 45 viral proteins and five host proteins, is presented, which will facilitate our understanding of the molecular mechanisms of baculovirus infection.

  7. Putative phosphorylation sites on WCA domain of HA2 is essential for Helicoverpa armigera single nucleopolyhedrovirus replication.

    PubMed

    Lv, Yi-pin; Wang, Qian; Wu, Chun-chen; Pei, Rong-juan; Zhou, Yuan; Wang, Yun; Chen, Xin-wen

    2011-08-01

    Protein phosphorylation is one of the most common post-translational modification processes that play an essential role in regulating protein functionality. The Helicoverpa armigera single nucleopolyhedrovirus (HearNPV) orf2-encoded nucleocapsid protein HA2 participates in orchestration of virus-induced actin polymerization through its WCA domain, in which phosphorylation status are supposed to be critical in respect to actin polymerization. In the present study, two putative phosphorylation sites ((232)Thr and (250)Ser) and a highly conserved Serine ((245)Ser) on the WCA domain of HA2 were mutated, and their phenotypes were characterized by reintroducing the mutated HA2 into the HearNPV genome. Viral infectivity assays demonstrated that only the recombinant HearNPV bearing HA2 mutation at (245)Ser can produce infectious virions, both (232)Thr and (250)Ser mutations were lethal to the virus. However, actin polymerization assay demonstrated that all the three viruses bearing HA2 mutations were still capable of initiating actin polymerization in the host nucleus, which indicated the putative phosphorylation sites on HA2 may contribute to HearNPV replication through another unidentified pathway.

  8. Inactivation of baculovirus by isoflavonoids on chickpea (Cicer arietinum) leaf surfaces reduces the efficacy of nucleopolyhedrovirus against Helicoverpa armigera.

    PubMed

    Stevenson, Philip C; D'Cunha, Reju F; Grzywacz, David

    2010-02-01

    Biological pesticides based on nucleopolyhedroviruses (NPVs) can provide an effective and environmentally benign alternative to synthetic chemicals. On some crops, however, the efficacy and persistence of NPVs is known to be reduced by plant specific factors. The present study investigated the efficacy of Helicoverpa armigera NPV (HearNPV) for control of H. armigera larvae, and showed that chickpea reduced the infectivity of virus occlusion bodies (OBs) exposed to the leaf surface of chickpea for at least 1 h. The degree of inactivation was greater on chickpea than that previously reported on cotton, and the mode of action is different from that of cotton. The effect was observed for larvae that consumed OBs on chickpea leaves, but it also occurred when OBs were removed after exposure to plants and inoculated onto artificial diet, indicating that inhibition was leaf surface-related and permanent. Despite their profuse exudation from trichomes on chickpea leaves and their low pH, organic acids-primarily oxalic and malic acid-caused no inhibition. When HearNPV was incubated with biochanin A and sissotrin, however, two minor constituents of chickpea leaf extracts, OB activity was reduced significantly. These two isoflavonoids increased in concentration by up to 3 times within 1 h of spraying the virus suspension onto the plants and also when spraying only the carrier, indicating induction was in response to spraying and not a specific response to the HearNPV. Although inactivation by the isoflavonoids did not account completely for the level of effect recorded on whole plants, this work constitutes evidence for a novel mechanism of NPV inactivation in legumes. Expanding the use of biological pesticides on legume crops will be dependent upon the development of suitable formulations for OBs to overcome plant secondary chemical effects.

  9. Dynamics of the interaction between cotton bollworm Helicoverpa armigera and nucleopolyhedrovirus as revealed by integrated transcriptomic and proteomic analyses.

    PubMed

    Xing, Longsheng; Yuan, Chuanfei; Wang, Manli; Lin, Zhe; Shen, Benchang; Hu, Zhihong; Zou, Zhen

    2017-04-12

    Over the past decades, Helicoverpa armigera nucleopolyhedrovirus (HearNPV) has been widely used for biocontrol of cotton bollworm, which is one of the most destructive pest insects in agriculture worldwide. However, the molecular mechanism underlying the interaction between HearNPV and host insects remains poorly understood. In this study, high throughput RNA-sequencing was integrated with label-free quantitative proteomics analysis to examine the dynamics of gene expression in the fat body of H. armigera larvae in response to challenge with HearNPV. RNA-sequencing-based transcriptomic analysis indicated that host gene expression was substantially altered, yielding 3,850 differentially expressed genes (DEGs), while no global transcriptional shut-off effects were observed in the fat body. Among the DEGs, 60 immunity-related genes were down-regulated after baculovirus infection, a finding that was consistent with the results of quantitative real-time RT-PCR (qRT-PCR). Gene ontology and functional classification demonstrated that the majority of down-regulated genes were enriched in gene cohorts involved in energy, carbohydrate, and amino acid metabolic pathways. Proteomics analysis identified differentially expressed proteins in the fat body, among which 76 were up-regulated, whereas 373 were significantly down-regulated upon infection. The down-regulated proteins are involved in metabolic pathways such as energy metabolism, carbohydrate metabolism (CM), and amino acid metabolism, in agreement with the RNA-seq data. Furthermore, correlation analysis suggested a strong association between the mRNA level and protein abundance in the H. armigera fat body. More importantly, the predicted gene interaction network indicated that a large subset of metabolic networks was significantly negatively regulated by viral infection, including CM-related enzymes such as aldolase, enolase, malate dehydrogenase, and triose-phosphate isomerase. Taken together, transcriptomic combined

  10. A Novel Binary Mixture of Helicoverpa armigera Single Nucleopolyhedrovirus Genotypic Variants Has Improved Insecticidal Characteristics for Control of Cotton Bollworms

    PubMed Central

    Arrizubieta, Maite; Simón, Oihane; Williams, Trevor

    2015-01-01

    The genotypic diversity of two Spanish isolates of Helicoverpa armigera single nucleopolyhedrovirus (HearSNPV) was evaluated with the aim of identifying mixtures of genotypes with improved insecticidal characteristics for control of the cotton bollworm. Two genotypic variants, HearSP1A and HearSP1B, were cloned in vitro from the most pathogenic wild-type isolate of the Iberian Peninsula, HearSNPV-SP1 (HearSP1-wt). Similarly, six genotypic variants (HearLB1 to -6) were obtained by endpoint dilution from larvae collected from cotton crops in southern Spain that died from virus disease during laboratory rearing. Variants differed significantly in their insecticidal properties, pathogenicity, speed of kill, and occlusion body (OB) production (OBs/larva). HearSP1B was ∼3-fold more pathogenic than HearSP1-wt and the other variants. HearLB1, HearLB2, HeaLB5, and HearLB6 were the fastest-killing variants. Moreover, although highly virulent, HearLB1, HearLB4, and HearLB5 produced more OBs/larva than did the other variants. The co-occluded HearSP1B:LB6 mixture at a 1:1 proportion was 1.7- to 2.8-fold more pathogenic than any single variant and other mixtures tested and also killed larvae as fast as the most virulent genotypes. Serial passage resulted in modified proportions of the component variants of the HearSP1B:LB6 co-occluded mixture, suggesting that transmissibility could be further improved by this process. We conclude that the improved insecticidal phenotype of the HearSP1B:LB6 co-occluded mixture underlines the utility of the genotypic variant dissection and reassociation approach for the development of effective virus-based insecticides. PMID:25841011

  11. A Novel Binary Mixture of Helicoverpa armigera Single Nucleopolyhedrovirus Genotypic Variants Has Improved Insecticidal Characteristics for Control of Cotton Bollworms.

    PubMed

    Arrizubieta, Maite; Simón, Oihane; Williams, Trevor; Caballero, Primitivo

    2015-06-15

    The genotypic diversity of two Spanish isolates of Helicoverpa armigera single nucleopolyhedrovirus (HearSNPV) was evaluated with the aim of identifying mixtures of genotypes with improved insecticidal characteristics for control of the cotton bollworm. Two genotypic variants, HearSP1A and HearSP1B, were cloned in vitro from the most pathogenic wild-type isolate of the Iberian Peninsula, HearSNPV-SP1 (HearSP1-wt). Similarly, six genotypic variants (HearLB1 to -6) were obtained by endpoint dilution from larvae collected from cotton crops in southern Spain that died from virus disease during laboratory rearing. Variants differed significantly in their insecticidal properties, pathogenicity, speed of kill, and occlusion body (OB) production (OBs/larva). HearSP1B was ∼3-fold more pathogenic than HearSP1-wt and the other variants. HearLB1, HearLB2, HeaLB5, and HearLB6 were the fastest-killing variants. Moreover, although highly virulent, HearLB1, HearLB4, and HearLB5 produced more OBs/larva than did the other variants. The co-occluded HearSP1B:LB6 mixture at a 1:1 proportion was 1.7- to 2.8-fold more pathogenic than any single variant and other mixtures tested and also killed larvae as fast as the most virulent genotypes. Serial passage resulted in modified proportions of the component variants of the HearSP1B:LB6 co-occluded mixture, suggesting that transmissibility could be further improved by this process. We conclude that the improved insecticidal phenotype of the HearSP1B:LB6 co-occluded mixture underlines the utility of the genotypic variant dissection and reassociation approach for the development of effective virus-based insecticides.

  12. Mutational and functional analysis of N-linked glycosylation of envelope fusion protein F of Helicoverpa armigera nucleopolyhedrovirus.

    PubMed

    Shen, Shu; Wang, Manli; Li, Xin; Li, Shufen; van Oers, Monique M; Vlak, Just M; Braakman, Ineke; Hu, Zhihong; Deng, Fei; Wang, Hualin

    2016-04-01

    The envelope fusion (F) protein of baculoviruses is a heavily N-glycosylated protein that plays a significant role in the virus infection cycle. N-Linked glycosylation of virus envelope glycoprotein is important for virus envelope glycoprotein folding and its function in general. There are six predicted N-glycosylation sites in the F (HaF) protein of Helicoverpa armigera nucleopolyhedrovirus (HearNPV). The N-glycosylation site located in the F(2) subunit (N104) of HaF has been identified and functionally characterized previously (Long et al., 2007). In this study, the other five potential N-glycosylation sites located in the HaF1 subunit, namely, N293, N361, N526, N571 and N595, were analysed extensively to examine their N-glycosylation and relative importance to the function of HaF. The results showed that four of these five potential glycosylation sites in the F(1) subunit, N293, N361, N526 and N571, were N-glycosylated in F proteins of mature HearNPV budded viruses (BVs) but that N595 was not. In general, the conserved site N526 was critical to the functioning of HaF, as absence of N-glycosylation of N526 reduced the efficiency of HaF folding and trafficking, consequently decreased fusogenicity and modified the subcellular localization of HaF proteins, and thus impaired virus production and infectivity. The absence of N-glycosylation at other individual sites was found to have different effects on the fusogenicity and subcelluar distribution of HaF proteins in HzAM1 cells. In summary, N-glycosylation plays comprehensive roles in HaF function and virus infectivity, which is further discussed.

  13. Mutagenesis and nuclear magnetic resonance analyses of the fusion peptide of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus F protein.

    PubMed

    Tan, Ying; Jiang, Ling; Wang, Manli; Yin, Feifei; Deng, Fei; Liu, Maili; Hu, Zhihong; Wang, Hualin

    2008-08-01

    The entry of enveloped viruses into cells is normally mediated by fusion between viral and cellular membranes, in which the fusion peptide plays a crucial role. The fusion peptides of group II nucleopolyhedrovirus (NPV) F proteins are quite conserved, with a hydrophobic region located at the N terminal of the F(1) fragment. For this report, we used mutagenesis and nuclear magnetic resonance (NMR) to study the structure and function of the fusion peptide of the Helicoverpa armigera single-nucleocapsid NPV (HearNPV) F protein (HaF). Five mutations in the fusion peptide of HaF, N(1)G, N(1)L, I(2)N, G(3)L, and D(11)L, were generated separately, and the mutated f genes were transformed into the f-null HearNPV bacmid. The mutations N(1)L, I(2)N, and D(11)L were found to completely abolish the ability of the recombinant bacmids to produce infectious budded virus, while the mutations N(1)G and G(3)L did not. The low-pH-induced envelope fusion assay demonstrated that the N(1)G substitution increased the fusogenicity of HaF, while the G(3)L substitution reduced its fusogenicity. NMR spectroscopy was used to determine the structure of a synthetic fusion peptide of HaF in the presence of sodium dodecyl sulfate micelles at pH 5.0. The fusion peptide appeared to be an amphiphilic structure composed of a flexible coil in the N terminus from N(1) to N(5), a 3(10)-helix from F(6) to G(8), a turn at S(9), and a regular alpha-helix from V(10) to D(19). The data provide the first NMR structure of a baculovirus fusion peptide and allow us to further understand the relationship of structure and function of the fusion peptide.

  14. Whole genome sequence and comparative genomic sequence analysis of Helicoverpa armigera nucleopolyhedrovirus (HearNPV-L1) isolated from India.

    PubMed

    Raghavendra, Ashika T; Jalali, Sushil K; Ojha, Rakshit; Shivalingaswamy, Timalapur M; Bhatnagar, Raj

    2017-03-01

    The whole genome of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) from India, HearNPV-L1, was sequenced and analyzed, with a view to look for genes and/or nucleotide sequences that might be involved in the differences and virulence among other HearNPVs sequenced from other countries like SP1A (Spain), NNg1 (Kenya) and G4 (China). The entire nucleotide sequence of the HearNPV-L1 genome was 136,740 bp in length having GC content of 39.19% and contained 113 ORFs that could encode polypeptides with more than 50 amino acids (GenBank accession number KT013224). Two ORFs, viz., ORF 18 (300 bp) and ORF 19 (401 bp) identified were unique in HearNPV-L1 genome. Most of the HearNPV-L1 ORFs showed high similarity to NNg1, SP1A and G4 genomes. HearNPV-L1 genome contains 5 h (hr1-hr5), these regions were found 84-100% similar to hr region of NNg1, SP1A and G4 genomes. A total of four bro genes were observed in HearNPV-L1 genome, of which bro-a gene was 12 and 351 bp bigger than SP1A and G4 bro-a, respectively, while bro-b was 15 bp bigger SP1A and NNg1 bro-b, whereas 593 bp shorter than G4 bro-b, while bro-c was 12 bp shorter than NNg1, however bro-c was absent in G4 genome. HearNPV-L1 bro-d was 100% homologous to bro-d of SP1A, NNg1 and G4 genomes, respectively. The comparative analysis of HearNPV-L1 genome indicated that there are several other putative genes and nucleotide sequences that may be responsible for insecticidal activity in HearNPV-L1 isolate, however, further functional analysis of the hypothetical (putative) genes may help identifying the genes that are crucial for the virulence and insecticidal activity.

  15. A Novel Neurotoxin Gene ar1b Recombination Enhances the Efficiency of Helicoverpa armigera Nucleopolyhedrovirus as a Pesticide by Inhibiting the Host Larvae Ability to Feed and Grow

    PubMed Central

    Yu, Huan; Meng, Jiao; Xu, Jian; Liu, Tong-xian; Wang, Dun

    2015-01-01

    A recombinant Helicoverpa armigera nucleopolyhedrovirus (HearNPV), Ar1b-HearNPV, was constructed and identified as an improved bio-control agent of Helicoverpa armigera larvae. The HearNPV polyhedrin promoter was used to express the insect-specific neurotoxin gene, ar1b, which was originally isolated from the Australian funnel-web spider (Atrax robustus). RT-PCR and Western blotting analysis showed that both the ar1b transcript and protein were produced successfully in Ar1b-HearNPV-infected HzAM1 cells. In order to investigate the influence of foreign gene insertion in HearNPV, including the ar1b gene, chloramphenicol resistance gene, lacZ, kanamycin resistance gene, and the gentamicin resistance gene, two virus strains (HZ8-HearNPV and wt-HearNPV) were used as controls in the cell transfection analysis. As expected, foreign gene insertion had no impact on budded virus production and viral DNA replication. Both optical microscopy and electron microscopy observations indicated that the formation of the occlusion bodies of recombinant virus was similar to wild type virus. The Ar1b-HearNPV-infected H. armigera larvae exhibited paralysis and weight loss before dying. This recombinant virus also showed a 32.87% decrease in LT50 assays compared with the wild type virus. Besides, Ar1b-HearNPV also inhibited host larval growth and diet consumption. This inhibition was still significant in the older instar larvae treated with the recombinant virus. All of these positive properties of this novel recombinant HearNPV provide a further opportunity to develop this virus strain into a commercial product to control the cotton bollworm. PMID:26296090

  16. A Novel Neurotoxin Gene ar1b Recombination Enhances the Efficiency of Helicoverpa armigera Nucleopolyhedrovirus as a Pesticide by Inhibiting the Host Larvae Ability to Feed and Grow.

    PubMed

    Yu, Huan; Meng, Jiao; Xu, Jian; Liu, Tong-Xian; Wang, Dun

    2015-01-01

    A recombinant Helicoverpa armigera nucleopolyhedrovirus (HearNPV), Ar1b-HearNPV, was constructed and identified as an improved bio-control agent of Helicoverpa armigera larvae. The HearNPV polyhedrin promoter was used to express the insect-specific neurotoxin gene, ar1b, which was originally isolated from the Australian funnel-web spider (Atrax robustus). RT-PCR and Western blotting analysis showed that both the ar1b transcript and protein were produced successfully in Ar1b-HearNPV-infected HzAM1 cells. In order to investigate the influence of foreign gene insertion in HearNPV, including the ar1b gene, chloramphenicol resistance gene, lacZ, kanamycin resistance gene, and the gentamicin resistance gene, two virus strains (HZ8-HearNPV and wt-HearNPV) were used as controls in the cell transfection analysis. As expected, foreign gene insertion had no impact on budded virus production and viral DNA replication. Both optical microscopy and electron microscopy observations indicated that the formation of the occlusion bodies of recombinant virus was similar to wild type virus. The Ar1b-HearNPV-infected H. armigera larvae exhibited paralysis and weight loss before dying. This recombinant virus also showed a 32.87% decrease in LT50 assays compared with the wild type virus. Besides, Ar1b-HearNPV also inhibited host larval growth and diet consumption. This inhibition was still significant in the older instar larvae treated with the recombinant virus. All of these positive properties of this novel recombinant HearNPV provide a further opportunity to develop this virus strain into a commercial product to control the cotton bollworm.

  17. Characterization of two monoclonal antibodies, 38F10 and 44D11, against the major envelope fusion protein of Helicoverpa armigera nucleopolyhedrovirus.

    PubMed

    Zou, Zijiao; Liu, Jinliang; Wang, Zhiying; Deng, Fei; Wang, Hualin; Hu, Zhihong; Wang, Manli; Zhang, Tao

    2016-12-01

    The envelope fusion protein F of baculoviruses is a class I viral fusion protein which play a significant role during virus entry into insect cells. F is initially synthesized as a precursor (F0) and then cleaved into a disulfide-linked F1 and F2 subunits during the process of protein maturation and secretion. To facilitate further investigation into the structure and function of F protein during virus infection, monoclonal antibodies (mAbs) against the F2 subunit of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) (HaF) were generated. Two kinds of mAbs were obtained according to their different recognition epitopes: one kind of mAbs, as represented by 38F10, recognizes amino acid (aa) 85 to 123 of F2 and the other kind, represented by 44D11, recognizes aa 148 to 173 of F2. Western blot and immunofluorescence assay confirmed that both of the mAbs recognized the F protein expressed in HearNPV infected cells, however, only 44D11 could neutralize HearNPV infection. The results further showed that 44D11 may not interact with a receptor binding epitope, rather it was demonstrated to inhibit syncytium formation in cells expressing the HaF protein. The results imply that the monoclonal antibody 44D11 recognizes a region within HaF2 that may be involved in the F-mediated membrane fusion process.

  18. Open reading frame 94 of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus encodes a novel conserved occlusion-derived virion protein, ODV-EC43.

    PubMed

    Fang, Minggang; Wang, Hanzhong; Wang, Hualin; Yuan, Li; Chen, Xinwen; Vlak, Just M; Hu, Zhihong

    2003-11-01

    Open reading frame 94 (Ha94) of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV) is 1086 bp long and a homologue of Autographa californica multiple NPV ORF109. The gene is conserved among all baculoviruses whose genomes have been completely sequenced so far and is thus considered a baculovirus core gene. Ha94 transcripts were detected from 24 to 96 h post-infection (p.i.) of HzAM1 cells with HaSNPV. Polyclonal antiserum raised to a GST-HA94 fusion protein recognized a 43 kDa protein, HA94, in infected cell lysates from 36 to 96 h p.i., suggesting that Ha94 is a late gene. Western blot analysis of proteins present in budded virus and occlusion-derived virus (ODV) showed that Ha94 encodes a structural component of ODV. When ODVs were fractionated further into nucleocapsid and envelope components, Western blot analysis indicated that the encoded protein was associated with both the nucleocapsid and the envelope. In summary, data available indicated that Ha94 encodes a novel ODV-specific protein of HaSNPV, designated ODV-EC43.

  19. Proteomics analysis of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus identified two new occlusion-derived virus-associated proteins, HA44 and HA100.

    PubMed

    Deng, Fei; Wang, Ranran; Fang, Minggang; Jiang, Yue; Xu, Xushi; Wang, Hanzhong; Chen, Xinwen; Arif, Basil M; Guo, Lin; Wang, Hualin; Hu, Zhihong

    2007-09-01

    Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry were used to analyze the structural proteins of the occlusion-derived virus (ODV) of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV), a group II NPV. Twenty-three structural proteins of HearNPV ODV were identified, 21 of which have been reported previously as structural proteins or ODV-associated proteins in other baculoviruses. These include polyhedrin, P78/83, P49, ODV-E18, ODV-EC27, ODV-E56, P74, LEF-3, HA66 (AC66), DNA polymerase, GP41, VP39, P33, ODV-E25, helicase, P6.9, ODV/BV-C42, VP80, ODV-EC43, ODV-E66, and PIF-1. Two proteins encoded by HearNPV ORF44 (ha44) and ORF100 (ha100) were discovered as ODV-associated proteins for the first time. ha44 encodes a protein of 378 aa with a predicted mass of 42.8 kDa. ha100 encodes a protein of 510 aa with a predicted mass of 58.1 kDa and is a homologue of the gene for poly(ADP-ribose) glycohydrolase (parg). Western blot analysis and immunoelectron microscopy confirmed that HA44 is associated with the nucleocapsid and HA100 is associated with both the nucleocapsid and the envelope of HearNPV ODV. HA44 is conserved in group II NPVs and granuloviruses but does not exist in group I NPVs, while HA100 is conserved only in group II NPVs.

  20. Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus ORF51 is a ChaB homologous gene involved in budded virus production and DNA replication.

    PubMed

    Zheng, Fangliang; Huang, Yi; Long, Gang; Sun, Xiulian; Wang, Hanzhong

    2011-01-01

    The baculovirus ChaB proteins are conserved in all completely sequenced Lepidopteran NPVs and are annotated as putative DNA binding proteins. Here we investigated Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) ORF51 (ha51), one of the ChaB homologues in HearNPV. 5'-RACE revealed that Ha51 is transcribed from a conventional early promoter transcriptional initiator motif (CATT) located at 159nt upstream of ATG. RT-PCR confirmed that ha51 is an early transcribed gene. To study the function of Ha51 in the life cycle of HearNPV, Ha51 knockout and repair bacmids were generated by homologous recombination in Escherichia coli. Growth curve and DNA replication analyses showed that the levels of budded virus (BV) production and viral DNA accumulation were significantly higher in cells infected with Ha51 null virus than those infected with wild-type bacmid derived virus. Electron microscopy revealed that polyhedra formation was not affected by the deletion of Ha51. Bioassay demonstrated that the Ha51-deleted virus had similar oral infectivity as the wild-type and rescued virus. Western blot analyses suggested that HA51 is a component of the nucleocapsid of BV and occlusion-derived virus as well as the envelope of BV. Immunofluorescence microscopy showed that HA51 protein is mainly localized in the cytoplasm of infected cells. Taken together, our results indicate that, unlike previously characterized baculovirual ChaB genes, Ha51 is involved in viral DNA replication and BV production and is transcribed in the early stage of infection.

  1. Ha83, a Chitin Binding Domain Encoding Gene, Is Important to Helicoverpa armigera Nucleopolyhedrovirus Budded Virus Production and Occlusion Body Assembling

    PubMed Central

    Yu, Huan; Xu, Jian; Liu, Qiang; Liu, Tong-Xian; Wang, Dun

    2015-01-01

    Helicoerpa armigera nucleopolyhedrovirus (HearNPV) ha83 is a late expressed gene that encodes a chitin binding protein. Chitin domain truncation studies revealed that the cysteine at the 128 amino acid position probably played an important role in both chitin binding ability and protein transmission of Ha83. In order to study the function of ha83 in the HearNPV infection cycle, an ha83 knockout HearNPV (Ha83KO) was constructed via homologous recombination. Viral growth and viral DNA replication curves showed that fewer budded virions were produced in Ha83KO transfected cells, while viral DNA replication was increased. Electron microscopy revealed that fewer nucleocapsids were transmitted from virogenic stroma in the Ha83KO transfected cell nucleus, and the morphology of occlusion bodies was prominently larger and cube-shaped. Furthermore, DNA quantity in occlusion bodies of Ha83KO was significantly lower than the occlusion bodies of HaWT. The transcription analysis indicated that these changes may be due to the decreased expression level of viral structural associated genes, such as polyhedrin, p10, pif-2, or cg30 in Ha83KO infected cells. Above results demonstrated that the cysteine at the 128 amino acid position in Ha83 might be the key amino acid, and Ha83 plays an important role in BVs production and OBs assembling. PMID:26057202

  2. Emamectin benzoate: new insecticide against Helicoverpa armigera.

    PubMed

    Fanigliulo, A; Sacchetti, M

    2008-01-01

    Emamectin benzoate is a new insecticide of Syngenta Crop Protection, with a new mechanism of action and a strong activity against Lepidoptera as well as with and a high selectivity on useful organisms. This molecule acts if swallowed and has some contact action. It penetrates leaf tissues (translaminar activity) and forms a reservoir within the leaf. The mechanism of action is unique in the panorama of insecticides. In facts, it inhibits muscle contraction, causing a continuous flow of chlorine ions in the GABA and H-Glutamate receptor sites. During 2006 and 2007, experimentation was performed by the Bioagritest test facility, according to EPPO guidelines and Principles of Good Experimental Practice (GEP), aiming at establishing the biological efficacy and the selectivity of Emamectin benzoate on industry tomato against Helicoverpa armigera (Lepidoptera: Noctuidoe). The study was performed in Tursi-Policoro (Matera), southern Italy. Experimental design consisted in random blocks, in 4 repetitions. A dosage of 1.5 Kg/ha of the formulate was compared with two commercial formulates: Spinosad 0.2 kg/ha (Laser, Dow Agrosciences Italia) and Indoxacarb 0.125 kg/ha (Steward EC insecticide, Dupont). Three foliage applications were applied every 8 days. The severity of damage induced by H. armigera was evaluated on fruits. Eventual phytotoxic effects were also evaluated. Climatic conditions were optimal for Lepidoptera development, so that the percentage of fruits attacked in 2007 at the first scouting was 68.28%. Emamectin benzoate has shown, in two years of testing, a high control of H. armigera if compared with the standards Indoxacarb and Spinosad. No effect of phytotoxicity was noticed on fruits.

  3. Battle in the New World: Helicoverpa armigera versus Helicoverpa zea (Lepidoptera: Noctuidae)

    PubMed Central

    2016-01-01

    The corn earworm Helicoverpa zea (Boddie) and the old world bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) are allopatric species and occur in important agricultural crops. In maize, both species tend to infest the ear. The introduction of H. armigera in Brazil has created a new scenario, where these Helicoverpa species might cohabit and interact with one another, affecting the prevalence of each species in the agroecosystem, integrated pest management, and insect resistance management. In this study, larval occurrence and proportion of these species in maize was assessed in three regions of Brazil during three crop seasons. Interaction between the species was evaluated in interspecific and intraspecific scenarios under laboratory and field conditions. Helicoverpa zea was predominant in Rio Grande do Sul and the Planaltina, DF (central Brazil). In western Bahia, H. zea was predominant in the first collection, but approximately equal in number to H armigera in the second crop season. Both species exhibit high cannibalism/predation rates, and larval size was the primary factor for larval survival in the interaction studies. Larva of H. zea had higher survival when interacting with H. armigera, indicating that H. zea has an advantage in intraguild interactions with H. armigera in maize. Overall, the results from this study indicate that maize might play a role as a source of infestation or a sink of insecticide or Bt protein unselected H. armigera populations, depending on the H. zea:H. armigera intraguild competition and adult movement in the landscape. PMID:27907051

  4. Battle in the New World: Helicoverpa armigera versus Helicoverpa zea (Lepidoptera: Noctuidae).

    PubMed

    Bentivenha, José P F; Paula-Moraes, Silvana V; Baldin, Edson L L; Specht, Alexandre; da Silva, Ivana F; Hunt, Thomas E

    2016-01-01

    The corn earworm Helicoverpa zea (Boddie) and the old world bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) are allopatric species and occur in important agricultural crops. In maize, both species tend to infest the ear. The introduction of H. armigera in Brazil has created a new scenario, where these Helicoverpa species might cohabit and interact with one another, affecting the prevalence of each species in the agroecosystem, integrated pest management, and insect resistance management. In this study, larval occurrence and proportion of these species in maize was assessed in three regions of Brazil during three crop seasons. Interaction between the species was evaluated in interspecific and intraspecific scenarios under laboratory and field conditions. Helicoverpa zea was predominant in Rio Grande do Sul and the Planaltina, DF (central Brazil). In western Bahia, H. zea was predominant in the first collection, but approximately equal in number to H armigera in the second crop season. Both species exhibit high cannibalism/predation rates, and larval size was the primary factor for larval survival in the interaction studies. Larva of H. zea had higher survival when interacting with H. armigera, indicating that H. zea has an advantage in intraguild interactions with H. armigera in maize. Overall, the results from this study indicate that maize might play a role as a source of infestation or a sink of insecticide or Bt protein unselected H. armigera populations, depending on the H. zea:H. armigera intraguild competition and adult movement in the landscape.

  5. Proteomic analysis of novel Cry1Ac binding proteins in Helicoverpa armigera (Hübner)

    USDA-ARS?s Scientific Manuscript database

    Aminopeptidase N (APN) and cadherin-like proteins have been previously identified as Cry1Ac-binding proteins in Helicoverpa armigera (Hübner). In this study, a proteomic approach was used to identify novel Cry1Ac-binding proteins in H. armigera. Brush border membrane vesicles (BBMV) of H. armigera w...

  6. The Spread of Helicoverpa armigera (Lepidoptera: Noctuidae) and Coexistence with Helicoverpa zea in Southeastern Brazil

    PubMed Central

    Pinto, Fábio A.; Mattos, Marcos V. V.; Rocha, Silma L.; Elliot, Simon L.

    2017-01-01

    Helicoverpa armigera, one of the world’s most destructive crop pests, was first documented in Brazil in 2013. Within a few months, this polyphagous insect had spread over the Northeast and Central-West of Brazil, causing great agricultural losses. With several reports of populations resistant to pesticides and Bt crops around the world, there is great concern about the spread of this pest in Brazil. There is confusion about the actual distribution of this species due to the high morphological similarity with the native corn earworm Helicoverpa zea, which may also coexist with H. armigera in the field. Our aims here were (i) to confirm its presence in the State of Minas Gerais, one of the most important agricultural regions in the country; and (ii) to assess the co-occurrence of this pest with the congeneric corn earworm H. zea. Using molecular screening, we confirmed the presence of H. armigera in Bt-crops of soybean and cotton, and non-Bt-crops of soybean, cotton and maize. Mixed infestations of H. armigera with H. zea were found in non-Bt maize (Viçosa, Southeastern Minas Gerais). These results highlight the need for adequate control strategies for H. armigera in Brazil, to deal with its polyphagous feeding habits, high dispersal capacity and possible risks of hybridization with congeneric species. PMID:28869528

  7. The Spread of Helicoverpa armigera (Lepidoptera: Noctuidae) and Coexistence with Helicoverpa zea in Southeastern Brazil.

    PubMed

    Pinto, Fábio A; Mattos, Marcos V V; Silva, Farley W S; Rocha, Silma L; Elliot, Simon L

    2017-09-04

    Helicoverpa armigera, one of the world's most destructive crop pests, was first documented in Brazil in 2013. Within a few months, this polyphagous insect had spread over the Northeast and Central-West of Brazil, causing great agricultural losses. With several reports of populations resistant to pesticides and Bt crops around the world, there is great concern about the spread of this pest in Brazil. There is confusion about the actual distribution of this species due to the high morphological similarity with the native corn earworm Helicoverpa zea, which may also coexist with H. armigera in the field. Our aims here were (i) to confirm its presence in the State of Minas Gerais, one of the most important agricultural regions in the country; and (ii) to assess the co-occurrence of this pest with the congeneric corn earworm H. zea. Using molecular screening, we confirmed the presence of H. armigera in Bt-crops of soybean and cotton, and non-Bt-crops of soybean, cotton and maize. Mixed infestations of H. armigera with H. zea were found in non-Bt maize (Viçosa, Southeastern Minas Gerais). These results highlight the need for adequate control strategies for H. armigera in Brazil, to deal with its polyphagous feeding habits, high dispersal capacity and possible risks of hybridization with congeneric species.

  8. Host plant induced variation in gut bacteria of Helicoverpa armigera.

    PubMed

    Priya, Natarajan Gayatri; Ojha, Abhishek; Kajla, Mayur K; Raj, Anand; Rajagopal, Raman

    2012-01-01

    Helicoverpa are important polyphagous agricultural insect pests and they have a worldwide distribution. In this study, we report the bacterial community structure in the midgut of fifth instar larvae of Helicoverpa armigera, a species prevalent in the India, China, South Asia, South East Asia, Southern & Eastern Africa and Australia. Using culturable techniques, we isolated and identified members of Bacillus firmus, Bacillus niabense, Paenibacillus jamilae, Cellulomonas variformis, Acinetobacter schindleri, Micrococcus yunnanesis, Enterobacter sp., and Enterococcus cassiliflavus in insect samples collected from host plants grown in different parts of India. Besides these the presence of Sphingomonas, Ralstonia, Delftia, Paracoccus and Bacteriodetes was determined by culture independent molecular analysis. We found that Enterobacter and Enterococcus were universally present in all our Helicoverpa samples collected from different crops and in different parts of India. The bacterial diversity varied greatly among insects that were from different host plants than those from the same host plant of different locations. This result suggested that the type of host plant greatly influences the midgut bacterial diversity of H. armigera, more than the location of the host plant. On further analyzing the leaf from which the larva was collected, it was found that the H. armigera midgut bacterial community was similar to that of the leaf phyllosphere. This finding indicates that the bacterial flora of the larval midgut is influenced by the leaf surface bacterial community of the crop on which it feeds. Additionally, we found that laboratory made media or the artificial diet is a poor bacterial source for these insects compared to a natural diet of crop plant.

  9. Host Plant Induced Variation in Gut Bacteria of Helicoverpa armigera

    PubMed Central

    Gayatri Priya, Natarajan; Ojha, Abhishek; Kajla, Mayur K.; Raj, Anand; Rajagopal, Raman

    2012-01-01

    Helicoverpa are important polyphagous agricultural insect pests and they have a worldwide distribution. In this study, we report the bacterial community structure in the midgut of fifth instar larvae of Helicoverpa armigera, a species prevalent in the India, China, South Asia, South East Asia, Southern & Eastern Africa and Australia. Using culturable techniques, we isolated and identified members of Bacillus firmus, Bacillus niabense, Paenibacillus jamilae, Cellulomonas variformis, Acinetobacter schindleri, Micrococcus yunnanesis, Enterobacter sp., and Enterococcus cassiliflavus in insect samples collected from host plants grown in different parts of India. Besides these the presence of Sphingomonas, Ralstonia, Delftia, Paracoccus and Bacteriodetes was determined by culture independent molecular analysis. We found that Enterobacter and Enterococcus were universally present in all our Helicoverpa samples collected from different crops and in different parts of India. The bacterial diversity varied greatly among insects that were from different host plants than those from the same host plant of different locations. This result suggested that the type of host plant greatly influences the midgut bacterial diversity of H. armigera, more than the location of the host plant. On further analyzing the leaf from which the larva was collected, it was found that the H. armigera midgut bacterial community was similar to that of the leaf phyllosphere. This finding indicates that the bacterial flora of the larval midgut is influenced by the leaf surface bacterial community of the crop on which it feeds. Additionally, we found that laboratory made media or the artificial diet is a poor bacterial source for these insects compared to a natural diet of crop plant. PMID:22292034

  10. Candidate olfaction genes identified within the Helicoverpa armigera Antennal Transcriptome.

    PubMed

    Liu, Yang; Gu, Shaohua; Zhang, Yongjun; Guo, Yuyuan; Wang, Guirong

    2012-01-01

    Antennal olfaction is extremely important for insect survival, mediating key behaviors such as host preference, mate choice, and oviposition site selection. Multiple antennal proteins are involved in olfactory signal transduction pathways. Of these, odorant receptors (ORs) and ionotropic receptors (IRs) confer specificity on olfactory sensory neuron responses. In this study, we identified the olfactory gene repertoire of the economically important agricultural pest moth, Helicoverpa armigera, by assembling the adult male and female antennal transcriptomes. Within the male and female antennal transcriptomes we identified a total of 47 OR candidate genes containing 6 pheromone receptor candidates. Additionally, 12 IR genes as well as 26 odorant-binding proteins and 12 chemosensory proteins were annotated. Our results allow a systematic functional analysis across much of conventional ORs repertoire and newly reported IRs mediating the key olfaction-mediated behaviors of H. armigera.

  11. Activity of Selected Formulated Biorational and Synthetic Insecticides Against Larvae of Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Vivan, L M; Torres, J B; Fernandes, P L S

    2016-12-23

    This work studied 17 insecticides belonging to nucleopolyhedrovirus (NPV), Bacillus thuringiensis (Bt kurstaki and Bt aizawai), benzoylureas (insect growth regulators [IGRs]), carbamates, organophosphates, spinosyns, and diamides against larvae of Helicoverpa armigera (Hübner), invasive species in the South American continent. Larvae of different instars were fed for 7 d with untreated or insecticide-treated diets. Mortality was recorded daily for 7 d, and surviving larvae were individually weighed on the seventh day. The NPV and Bt insecticides caused 100% mortality of first-instar larvae and first-instar and second-instar larvae, respectively. However, both NPV and Bt-based products caused low mortality of third-instar larvae and did not kill older larvae. The IGR lufenuron was highly effective against all three ages of larvae tested, whereas teflubenzuron and triflumuron produced maximum 60% mortality of second-instar larvae and lower than 50% to older larvae. Thiodicarb, chlorantraniliprole, indoxacarb, chlorpyrifos, and chlorfenapyr, irrespective of tested age, caused 100% mortality of larvae, with the last two insecticides reaching 100% mortality within 2 d of feeding on the treated diet. Flubendiamide caused lower mortality but significantly affected the weight of surviving larvae, whereas neither spinosad nor methomyl produced significant mortality or affected the weight of larvae. Based on the results, the age of H. armigera larvae plays an important role in the recommendation of NPV and Bt insecticides. Furthermore, there are potential options between biological and synthetic insecticides tested against H. armigera, and recording larval size during monitoring, in addition to the infestation level, should be considered when recommending biological-based insecticides to control this pest.

  12. Efficacy of Venom from Tentacle of Jellyfish Stomolophus meleagris (Nemopilema nomurai) against the Cotton Bollworm Helicoverpa armigera

    PubMed Central

    Yu, Huahua; Li, Rongfeng; Dong, Xiangli; Xing, Ronge; Liu, Song; Li, Pengcheng

    2014-01-01

    Efficacy of venom from tentacle of jellyfish Stomolophus meleagris against the cotton bollworm Helicoverpa armigera was determined. Venom from tentacle of jellyfish Stomolophus meleagris could inhibit the growth of Helicoverpa armigera and the weight inhibiting rate of sample NFr-2 was 60.53%. Of the six samples, only NFr-2 had high insecticidal activity against Helicoverpa armigera and the corrected mortality recorded at 7 d was 74.23%. PMID:25162008

  13. Olfactory perception and behavioral effects of sex pheromone gland components in Helicoverpa armigera and Helicoverpa assulta

    PubMed Central

    Xu, Meng; Guo, Hao; Hou, Chao; Wu, Han; Huang, Ling-Qiao; Wang, Chen-Zhu

    2016-01-01

    Two sympatric species Helicoverpa armigera and Helicoverpa assulta use (Z)-11-hexadecenal and (Z)-9-hexadecenal as sex pheromone components in reverse ratio. They also share several other pheromone gland components (PGCs). We present a comparative study on the olfactory coding mechanism and behavioral effects of these additional PGCs in pheromone communication of the two species using single sensillum recording, in situ hybridization, calcium imaging, and wind tunnel. We classify antennal sensilla types A, B and C into A, B1, B2, C1, C2 and C3 based on the response profiles, and identify the glomeruli responsible for antagonist detection in both species. The abundance of these sensilla types when compared with the number of OSNs expressing each of six pheromone receptors suggests that HarmOR13 and HassOR13 are expressed in OSNs housed within A type sensilla, HarmOR14b within B and C type sensilla, while HassOR6 and HassOR16 within some of C type sensilla. We find that for H. armigera, (Z)-11-hexadecenol and (Z)-11-hexadecenyl acetate act as behavioral antagonists. For H. assulta, instead, (Z)-11-hexadecenyl acetate acts as an agonist, while (Z)-9-hexadecenol, (Z)-11-hexadecenol and (Z)-9-hexadecenyl acetate are antagonists. The results provide an overall picture of intra- and interspecific olfactory and behavioral responses to all PGCs in two sister species. PMID:26975244

  14. Rapid identification of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) using ribosomal RNA internal transcribed spacer 1

    USDA-ARS?s Scientific Manuscript database

    Rapid identification of invasive species is crucial for deploying management strategies to prevent establishment. Recent Helicoverpa armigera (Hübner) invasions and subsequent establishment in some countries of South America has increased the risk of this species invading North America. Differentiat...

  15. Pan-American Similarities in Genetic Structures of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) With Implications for Hybridization.

    PubMed

    Leite, N A; Correa, A S; Michel, A P; Alves-Pereira, A; Pavinato, V A C; Zucchi, M I; Omoto, C

    2017-08-01

    The genus Helicoverpa (Lepidoptera: Noctuidae) includes phytophagous and polyphagous agricultural insect pests. In the Americas, a native pest, Helicoverpa zea (Boddie), and an invasive pest, Helicoverpa armigera (Hübner), are causing severe damage in vegetable and agronomic crops. The population structure of both species in South America is poorly understood, and the phylogenetic relatedness of H. armigera and H. zea suggests natural interspecific gene flow between these species. Using microsatellite loci, we investigated: 1) the genetic diversity and gene flow of H. armigera specimens from Brazil; 2) the genetic diversity and gene flow between H. zea specimens from Brazil and the United States; and 3) the possibility of interspecific gene flow and the frequency of putative hybrids in Brazil. We detected high intraspecific gene flow among populations collected in the same country. However, there is a geographic limit to gene flow among H. zea individuals from South and North America. Pairwise Fst and private alleles showed that H. armigera is more similar to H. zea from Brazil than H. zea from the United States. A comparative STRUCTURE analysis suggests natural hybridization between H. armigera and H. zea in Brazil. High gene flow and natural hybridization are key traits to population adaptation in new and disturbed environments, which can influence the management of these pests in the American continent. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  16. Carbon dioxide receptor genes in cotton bollworm Helicoverpa armigera

    NASA Astrophysics Data System (ADS)

    Xu, Wei; Anderson, Alisha

    2015-04-01

    Carbon dioxide (CO2) is important in insect ecology, eliciting a range of behaviours across different species. Interestingly, the numbers of CO2 gustatory receptors (GRs) vary among insect species. In the model organism Drosophila melanogaster, two GRs (DmelGR21a and DmelGR63a) have been shown to detect CO2. In the butterfly, moth, beetle and mosquito species studied so far, three CO2 GR genes have been identified, while in tsetse flies, four CO2 GR genes have been identified. In other species including honeybees, pea aphids, ants, locusts and wasps, no CO2 GR genes have been identified from the genome. These genomic differences may suggest different mechanisms for CO2 detection exist in different insects but, with the exception of Drosophila and mosquitoes, limited attention has been paid to the CO2 GRs in insects. Here, we cloned three putative CO2 GR genes from the cotton bollworm Helicoverpa armigera and performed phylogenetic and expression analysis. All three H. armigera CO2 GRs (HarmGR1, HarmGR2 and HarmGR3) are specifically expressed in labial palps, the CO2-sensing tissue of this moth. HarmGR3 is significantly activated by NaHCO3 when expressed in insect Sf9 cells but HarmGR1 and HarmGR2 are not. This is the first report characterizing the function of lepidopteran CO2 receptors, which contributes to our general understanding of the molecular mechanisms of insect CO2 gustatory receptors.

  17. Carbon dioxide receptor genes in cotton bollworm Helicoverpa armigera.

    PubMed

    Xu, Wei; Anderson, Alisha

    2015-04-01

    Carbon dioxide (CO2) is important in insect ecology, eliciting a range of behaviours across different species. Interestingly, the numbers of CO2 gustatory receptors (GRs) vary among insect species. In the model organism Drosophila melanogaster, two GRs (DmelGR21a and DmelGR63a) have been shown to detect CO2. In the butterfly, moth, beetle and mosquito species studied so far, three CO2 GR genes have been identified, while in tsetse flies, four CO2 GR genes have been identified. In other species including honeybees, pea aphids, ants, locusts and wasps, no CO2 GR genes have been identified from the genome. These genomic differences may suggest different mechanisms for CO2 detection exist in different insects but, with the exception of Drosophila and mosquitoes, limited attention has been paid to the CO2 GRs in insects. Here, we cloned three putative CO2 GR genes from the cotton bollworm Helicoverpa armigera and performed phylogenetic and expression analysis. All three H. armigera CO2 GRs (HarmGR1, HarmGR2 and HarmGR3) are specifically expressed in labial palps, the CO2-sensing tissue of this moth. HarmGR3 is significantly activated by NaHCO3 when expressed in insect Sf9 cells but HarmGR1 and HarmGR2 are not. This is the first report characterizing the function of lepidopteran CO2 receptors, which contributes to our general understanding of the molecular mechanisms of insect CO2 gustatory receptors.

  18. Demographics and Genetic Variability of the New World Bollworm (Helicoverpa zea) and the Old World Bollworm (Helicoverpa armigera) in Brazil

    PubMed Central

    Leite, Natália A.; Alves-Pereira, Alessandro; Corrêa, Alberto S.; Zucchi, Maria I.; Omoto, Celso

    2014-01-01

    Helicoverpa armigera is one of the primary agricultural pests in the Old World, whereas H. zea is predominant in the New World. However, H. armigera was first documented in Brazil in 2013. Therefore, the geographical distribution, range of hosts, invasion source, and dispersal routes for H. armigera are poorly understood or unknown in Brazil. In this study, we used a phylogeographic analysis of natural H. armigera and H. zea populations to (1) assess the occurrence of both species on different hosts; (2) infer the demographic parameters and genetic structure; (3) determine the potential invasion and dispersal routes for H. armigera within the Brazilian territory; and (4) infer the geographical origin of H. armigera. We analyzed partial sequence data from the cytochrome c oxidase subunit I (COI) gene. We determined that H. armigera individuals were most prevalent on dicotyledonous hosts and that H. zea were most prevalent on maize crops, based on the samples collected between May 2012 and April 2013. The populations of both species showed signs of demographic expansion, and no genetic structure. The high genetic diversity and wide distribution of H. armigera in mid-2012 are consistent with an invasion period prior to the first reports of this species in the literature and/or multiple invasion events within the Brazilian territory. It was not possible to infer the invasion and dispersal routes of H. armigera with this dataset. However, joint analyses using sequences from the Old World indicated the presence of Chinese, Indian, and European lineages within the Brazilian populations of H. armigera. These results suggest that sustainable management plans for the control of H. armigera will be challenging considering the high genetic diversity, polyphagous feeding habits, and great potential mobility of this pest on numerous hosts, which favor the adaptation of this insect to diverse environments and control strategies. PMID:25409452

  19. Demographics and genetic variability of the new world bollworm (Helicoverpa zea) and the old world bollworm (Helicoverpa armigera) in Brazil.

    PubMed

    Leite, Natália A; Alves-Pereira, Alessandro; Corrêa, Alberto S; Zucchi, Maria I; Omoto, Celso

    2014-01-01

    Helicoverpa armigera is one of the primary agricultural pests in the Old World, whereas H. zea is predominant in the New World. However, H. armigera was first documented in Brazil in 2013. Therefore, the geographical distribution, range of hosts, invasion source, and dispersal routes for H. armigera are poorly understood or unknown in Brazil. In this study, we used a phylogeographic analysis of natural H. armigera and H. zea populations to (1) assess the occurrence of both species on different hosts; (2) infer the demographic parameters and genetic structure; (3) determine the potential invasion and dispersal routes for H. armigera within the Brazilian territory; and (4) infer the geographical origin of H. armigera. We analyzed partial sequence data from the cytochrome c oxidase subunit I (COI) gene. We determined that H. armigera individuals were most prevalent on dicotyledonous hosts and that H. zea were most prevalent on maize crops, based on the samples collected between May 2012 and April 2013. The populations of both species showed signs of demographic expansion, and no genetic structure. The high genetic diversity and wide distribution of H. armigera in mid-2012 are consistent with an invasion period prior to the first reports of this species in the literature and/or multiple invasion events within the Brazilian territory. It was not possible to infer the invasion and dispersal routes of H. armigera with this dataset. However, joint analyses using sequences from the Old World indicated the presence of Chinese, Indian, and European lineages within the Brazilian populations of H. armigera. These results suggest that sustainable management plans for the control of H. armigera will be challenging considering the high genetic diversity, polyphagous feeding habits, and great potential mobility of this pest on numerous hosts, which favor the adaptation of this insect to diverse environments and control strategies.

  20. Diet-delivered RNAi in Helicoverpa armigera--Progresses and challenges.

    PubMed

    Lim, Zhi Xian; Robinson, Karl E; Jain, Ritesh G; Chandra, G Sharath; Asokan, R; Asgari, Sassan; Mitter, Neena

    2016-02-01

    Helicoverpa armigera (the cotton bollworm) is a significant agricultural pest endemic to Afro-Eurasia and Oceania. Gene suppression via RNA interference (RNAi) presents a potential avenue for management of the pest, which is highly resistant to traditional insecticide sprays. This article reviews current understanding on the fate of ingested double-stranded RNA in H. armigera. Existing in vivo studies on diet-delivered RNAi and their effects are summarized and followed by a discussion on the factors and hurdles affecting the efficacy of diet-delivered RNAi in H. armigera.

  1. Effects of a new microbial α-amylase inhibitor protein on Helicoverpa armigera larvae.

    PubMed

    Zeng, Fanrong; Wang, Xiaojing; Cui, Jinjie; Ma, Yan; Li, Qiannan

    2013-03-06

    A new microbial α-amylase inhibitor gene was cloned and characterized. The encoded, recombinant, α-amylase inhibitor protein was induced and expressed by isopropyl β-d-1-thiogalactopyranoside (IPTG) in Escherichia coli M15 cells. The effects of the α-amylase inhibitor protein on Helicoverpa armigera larvae were studied. Compared to the control, the weight of H. armigera larvae fed the diet with recombinant α-amylase inhibitor protein added at a concentration of 20 μg/g was reduced by 49.8%. The total soluble protein of H. armigera larvae fed the diet with the α-amylase inhibitor protein added was also reduced by 36.8% compared to the control. The recombinant α-amylase inhibitor protein showed inhibition activity against α-amylase of H. armigera. These results suggested that this α-amylase inhibitor protein may be a promising bioinsecticide candidate for controlling H. armigera.

  2. Intra- and extracellular domains of the Helicoverpa armigera cadherin mediate Cry1Ac cytotoxicity

    USDA-ARS?s Scientific Manuscript database

    Diverse midgut cadherin mutations confer resistance to Cry1A toxins in at least three lepidopteran pests, including the cotton bollworm, Helicoverpa armigera. Most of these cadherin mutations are inherited as recessive alleles and result in changes within the cadherin repeat (CR) regions of the extr...

  3. Prediction of cotton resistance to Helicoverpa armigera based on the percent (+)-gossypol in mature seed

    USDA-ARS?s Scientific Manuscript database

    Various Uzbek commercial varieties were grown in the field and these were exposed to cotton bollworm (Helicoverpa armigera) larvae. A significant negative correlation coefficient (r = -0.89) and linear regression (Y = 109.69-5.26X) was observed between the concentration of (+)-gossypol in cotton se...

  4. Rapid Identification of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) Using Ribosomal RNA Internal Transcribed Spacer 1

    PubMed Central

    Perera, Omaththage P.; Allen, Kerry C.; Jain, Devendra; Purcell, Matthew; Little, Nathan S.; Luttrell, Randall G.

    2015-01-01

    Rapid identification of invasive species is crucial for deploying management strategies to prevent establishment. Recent Helicoverpa armigera (Hübner) invasions and subsequent establishment in South America has increased the risk of this species invading North America. Morphological similarities make differentiation of H. armigera from the native Helicoverpa zea (Boddie) difficult. Characteristics of adult male genitalia and nucleotide sequence differences in mitochondrial DNA are two of the currently available methods to differentiate these two species. However, current methods are likely too slow to be employed as rapid detection methods. In this study, conserved differences in the internal transcribed spacer 1 (ITS1) of the ribosomal RNA genes were used to develop species-specific oligonucleotide primers that amplified ITS1 fragments of 147 and 334 bp from H. armigera and H. zea, respectively. An amplicon (83 bp) from a conserved region of 18S ribosomal RNA subunit served as a positive control. Melting temperature differences in ITS1 amplicons yielded species-specific dissociation curves that could be used in high resolution melt analysis to differentiate the two Helicoverpa species. In addition, a rapid and inexpensive procedure for obtaining amplifiable genomic DNA from a small amount of tissue was identified. Under optimal conditions, the process was able to detect DNA from one H. armigera leg in a pool of 25 legs. The high resolution melt analysis combined with rapid DNA extraction could be used as an inexpensive method to genetically differentiate large numbers of H. armigera and H. zea using readily available reagents. PMID:26516166

  5. Rapid identification of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) using ribosomal RNA internal transcribed spacer 1.

    PubMed

    Perera, Omaththage P; Allen, Kerry C; Jain, Devendra; Purcell, Matthew; Little, Nathan S; Luttrell, Randall G

    2015-01-01

    Rapid identification of invasive species is crucial for deploying management strategies to prevent establishment. Recent Helicoverpa armigera (Hübner) invasions and subsequent establishment in South America has increased the risk of this species invading North America. Morphological similarities make differentiation of H. armigera from the native Helicoverpa zea (Boddie) difficult. Characteristics of adult male genitalia and nucleotide sequence differences in mitochondrial DNA are two of the currently available methods to differentiate these two species. However, current methods are likely too slow to be employed as rapid detection methods. In this study, conserved differences in the internal transcribed spacer 1 (ITS1) of the ribosomal RNA genes were used to develop species-specific oligonucleotide primers that amplified ITS1 fragments of 147 and 334 bp from H. armigera and H. zea, respectively. An amplicon (83 bp) from a conserved region of 18S ribosomal RNA subunit served as a positive control. Melting temperature differences in ITS1 amplicons yielded species-specific dissociation curves that could be used in high resolution melt analysis to differentiate the two Helicoverpa species. In addition, a rapid and inexpensive procedure for obtaining amplifiable genomic DNA from a small amount of tissue was identified. Under optimal conditions, the process was able to detect DNA from one H. armigera leg in a pool of 25 legs. The high resolution melt analysis combined with rapid DNA extraction could be used as an inexpensive method to genetically differentiate large numbers of H. armigera and H. zea using readily available reagents.

  6. Population structure and gene flow in the global pest, Helicoverpa armigera.

    PubMed

    Anderson, C J; Tay, W T; McGaughran, A; Gordon, K; Walsh, T K

    2016-11-01

    Helicoverpa armigera is a major agricultural pest that is distributed across Europe, Asia, Africa and Australasia. This species is hypothesized to have spread to the Americas 1.5 million years ago, founding a population that is at present, a distinct species, Helicoverpa zea. In 2013, H. armigera was confirmed to have re-entered South America via Brazil and subsequently spread. The source of the recent incursion is unknown and population structure in H. armigera is poorly resolved, but a basic understanding would highlight potential biosecurity failures and determine the recent evolutionary history of region-specific lineages. Here, we integrate several end points derived from high-throughput sequencing to assess gene flow in H. armigera and H. zea from populations across six continents. We first assemble mitochondrial genomes to demonstrate the phylogenetic relationship of H. armigera with other Heliothine species and the lack of distinction between populations. We subsequently use de novo genotyping-by-sequencing and whole-genome sequences aligned to bacterial artificial chromosomes, to assess levels of admixture. Primarily, we find that Brazilian H. armigera are derived from diverse source populations, with strong signals of gene flow from European populations, as well as prevalent signals of Asian and African ancestry. We also demonstrate a potential field-caught hybrid between H. armigera and H. zea, and are able to provide genomic support for the presence of the H. armigera conferta subspecies in Australasia. While structure among the bulk of populations remains unresolved, we present distinctions that are pertinent to future investigations as well as to the biosecurity threat posed by H. armigera.

  7. Hybridization between Helicoverpa armigera and Helicoverpa assulta (Lepidoptera: Noctuidae): development and morphological characterization of F1 hybrids.

    PubMed

    Zhao, X-C; Dong, J-F; Tang, Q-B; Yan, Y-H; Gelbic, I; Van Loon, J J A; Wang, C-Z

    2005-10-01

    Reciprocal hybridizations between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) were studied. The cross between females of H. armigera and males of H. assulta yielded only fertile males and sterile individuals lacking an aedeagus, valva or ostium bursae. A total of 492 larvae of the F1 generation were obtained and 374 of these completed larval development and pupated. Only 203 pupae were morphologically normal males, the remaining 171 pupae were malformed. Larvae and pupae that gave rise to morphologically abnormal adults exhibited longer development times. Sterility was not only associated with malformed external sex organs, but also a range of abnormalities of the internal reproductive system: (i) loss of internal reproductive organs, (ii) with one to three copies of an undeveloped bursa copulatrix; or (iii) with one or two undeveloped testes. Normal male hybrid adults showed higher flight activity in comparison with males of both species. In contrast, the cross between females of H. assulta and males of H. armigera yielded morphologically normal offspring (80 males and 83 females). The interaction of the Z-chromosome from H. assulta with autosomes from H. armigera might result in morphological abnormalities found in hybrids and backcrosses, and maternal-zygotic incompatibilities might contribute to sex bias attributed to hybrid inviability.

  8. Responses of midgut amylases of Helicoverpa armigera to feeding on various host plants.

    PubMed

    Kotkar, Hemlata M; Sarate, Priya J; Tamhane, Vaijayanti A; Gupta, Vidya S; Giri, Ashok P

    2009-08-01

    Midgut digestive amylases and proteinases of Helicoverpa armigera, a polyphagous and devastating insect pest of economic importance have been studied. We also identified the potential of a sorghum amylase inhibitor against H. armigera midgut amylase. Amylase activities were detected in all the larval instars, pupae, moths and eggs; early instars had lower amylase levels which steadily increased up to the sixth larval instar. Qualitative and quantitative differences in midgut amylases of H. armigera upon feeding on natural and artificial diets were evident. Natural diets were categorized as one or more members of legumes, vegetables, flowers and cereals belonging to different plant families. Amylase activity and isoform patterns varied depending on host plant and/or artificial diet. Artificial diet-fed H. armigera larvae had comparatively high amylase activity and several unique amylase isoforms. Correlation of amylase and proteinase activities of H. armigera with the protein and carbohydrate content of various diets suggested that H. armigera regulates the levels of these digestive enzymes in response to macromolecular composition of the diet. These adjustments in the digestive enzymes of H. armigera may be to obtain better nourishment from the diet and avoid toxicity due to nutritional imbalance. H. armigera, a generalist feeder experiences a great degree of nutritional heterogeneity in its diet. An investigation of the differences in enzyme levels in response to macronutrient balance and imbalance highlight their importance in insect nutrition.

  9. Mitochondrial DNA COI characterization of Helicoverpa armigera (Lepidoptera: Noctuidae) from Paraguay and Uruguay.

    PubMed

    Arnemann, J A; James, W J; Walsh, T K; Guedes, J V C; Smagghe, G; Castiglioni, E; Tay, W T

    2016-04-07

    Since its detection in Brazil in 2013, the Old World cotton bollworm Helicoverpa armigera has been reported in Argentina, Paraguay, and Bolivia. Here we present evidence extending the South American range of H. armigera to Uruguay, using polymerase chain reaction and sequencing of the partial mitochondrial DNA (mtDNA) cytochrome oxidase I region. Molecular characterization of this gene region from individuals from Paraguay also supports previous morphological identification of H. armigera in Paraguay. Shared mtDNA haplotypes in H. armigera from Brazil, Uruguay, and Paraguay were identified. Additional surveying of populations in this region will be imperative to better monitor and understand factors that are underpinning its presence and successful adaptation in these South American regions. We discuss our findings with respect to the development of resistance pest management strategies of this invasive insect pest in a predominantly monoculture soybean crop landscape in the Southern Cone region.

  10. A droplet digital PCR (ddPCR) assay to detect Helicoverpa armigera (Lepidoptera: Noctuidae) in bulk trap samples

    USDA-ARS?s Scientific Manuscript database

    Moths in the genus Helicoverpa are some of the most important agricultural pests in the world. Two species, H. armigera (Hübner) and H. zea (Boddie), cause the majority of damage to crops and millions of dollars are spent annually on control of these pests. The recent introduction of H. armigera int...

  11. Developmental and Digestive Flexibilities in the Midgut of a Polyphagous Pest, the Cotton Bollworm, Helicoverpa armigera

    PubMed Central

    Sarate, P.J.; Tamhane, V.A.; Kotkar, H.M.; Ratnakaran, N.; Susan, N.; Gupta, V.S.; Giri, A.P.

    2012-01-01

    Developmental patterns and survival of the cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), a polyphagous insect pest, have been studied with reference to the effect of diet on major gut digestive enzymes (amylases, proteases, and lipases). Significant correlations between nutritional quality of the diet and larval and pupal mass were observed when H. armigera larvae were fed on various host plants viz. legumes (chickpea and pigeonpea), vegetables (tomato and okra), flowers (rose and marigold), and cereals (sorghum and maize). Larvae fed on diets rich in proteins and/or carbohydrates (pigeonpea, chickpea, maize, and sorghum) showed higher larval mass and developed more rapidly than larvae fed on diets with low protein and carbohydrate content (rose, marigold, okra, and tomato). Low calorific value diets like rose and marigold resulted in higher mortality (25–35%) of H. armigera. Even with highly varying development efficiency and larval/pupal survival rates, H. armigera populations feeding on different diets completed their life cycles. Digestive enzymes of H. armigera displayed variable expression levels and were found to be regulated on the basis of macromolecular composition of the diet. Post—ingestive adaptations operating at the gut level, in the form of controlled release of digestive enzymes, might be a key factor contributing to the physiological plasticity in H. armigera. PMID:22954360

  12. Seasonal migration of Helicoverpa armigera (Lepidoptera: Noctuidae) over the Bohai Sea.

    PubMed

    Feng, Hongqiang; Wu, Xianfu; Wu, Bo; Wu, Kongming

    2009-02-01

    The seasonal migration of the Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) over the Bohai Sea was observed with a searchlight trap and an entomological radar located on a small island in the center of the sea, and through a network of light-traps around the Bohai region. The H. armigera moths were observed to migrate over the sea at least as early as May and light trapping through a network suggested migration might start as early as April, as soon as the moths had emerged from overwintering pupae. H. armigera moths migrated toward the north in southerly winds during spring and summer, and returned south on nights with northerly winds, or at altitudes where the wind was northerly, during fall. The passage of a weather front (cold or warm) or trough at approximately 1700 hours provokes migration of H. armigera over the sea. The H. armigera generally flew at altitudes of below 1,500 m above sea level (asl) with layer concentrations at 200-500 m asl, where the wind direction, wind speed, and temperature were optimum. During fall migration, H. armigera tended to orient toward the southwest and was able to compensate for the wind drift by turning clockwise when the downwind direction was < 225 degrees but counterclockwise when it was > 225 degrees. The displacement speed measured with the radar was 24-41 km/h, the duration of flight was 8-11 h and the maximum migration rate was 1,894 moths per km per h.

  13. A Brave New World for an Old World Pest: Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil

    PubMed Central

    Walsh, Thomas; Thomazoni, Danielle; Silvie, Pierre; Behere, Gajanan T.; Anderson, Craig; Downes, Sharon

    2013-01-01

    The highly polyphagous Old World cotton bollworm Helicoverpa armigera is a quarantine agricultural pest for the American continents. Historically H. armigera is thought to have colonised the American continents around 1.5 to 2 million years ago, leading to the current H. zea populations on the American continents. The relatively recent species divergence history is evident in mating compatibility between H. zea and H. armigera under laboratory conditions. Despite periodic interceptions of H. armigera into North America, this pest species is not believed to have successfully established significant populations on either continent. In this study, we provide molecular evidence via mitochondrial DNA (mtDNA) cytochrome oxidase I (COI) and cytochrome b (Cyt b) partial gene sequences for the successful recent incursion of H. armigera into the New World, with individuals being detected at two sites (Primavera do Leste, Pedra Preta) within the State of Mato Grosso in Brazil. The mtDNA COI and Cyt b haplotypes detected in the Brazilian H. armigera individuals are common throughout the Old World, thus precluding identification of the founder populations. Combining the two partial mtDNA gene sequences showed that at least two matrilines are present in Brazil, while the inclusion of three nuclear DNA Exon-Primed Intron-Crossing (EPIC) markers identified a further two possible matrilines in our samples. The economic, biosecurity, resistance management, ecological and evolutionary implications of this incursion are discussed in relation to the current agricultural practices in the Americas. PMID:24260345

  14. Data of in vitro synthesized dsRNAs on growth and development of Helicoverpa armigera.

    PubMed

    Chikate, Yojana R; Dawkar, Vishal V; Barbole, Ranjit S; Tilak, Priyadarshini V; Gupta, Vidya S; Giri, Ashok P

    2016-06-01

    The data presented in this article is related to the research article "RNAi of selected candidate genes interrupts growth and development of Helicoverpa armigera" (Chikate et al., 2016) [1]. RNA interference (RNAi) is emerging as a potent insect pest control strategy over current methods and their resistance by pest. In this study we tested 15 different in vitro synthesized dsRNAs for gene silencing in Helicoverpa armigera. These dsRNAs were specific against H. armigera enzymes/proteins such as proteases like trypsins (HaTry2, 3, 4 and 6), chymotrypsin (HaChy4) and cysteine proteases such as cathepsin (HaCATHL); glutathione S-transferases (HaGST1a, 6 and 8); esterases (HaAce4, HaJHE); catalase (HaCAT); super-oxide-dismutase (HaCu/ZnSOD); fatty acid binding protein (HaFabp) and chitin deacetylase (HaCda5b). These dsRNAs were fed to second instar larvae at an optimized dose (60 µg/day) for 3 days separately. Effects of dsRNA feeding were observed in terms of larval mass gain, percentage mortality and phenotypic abnormalities in later developmental stages of H. armigera. These findings might provide potential new candidates for designing sequence-specific dsRNA as pesticide in crop protection.

  15. Comparative host selection responses of specialist (Helicoverpa assulta) and generalist (Helicoverpa armigera) moths in complex plant environments

    PubMed Central

    Li, Wei-zheng; Teng, Xiao-hui; Zhang, Hong-fei; Liu, Ting; Wang, Qiong; Guo, Xian-ru

    2017-01-01

    We tested the behavioral responses of ovipositing females and natal larvae of two sibling species, a generalist Helicoverpa armigera (Hübner) and a specialist Helicoverpa assulta (Guenée), to odor sources emitted from different combinations of six plant species (tobacco, Nicotiana tabacum; hot pepper, Capsicum annuum; tomato, Solanum esculentum; cotton, Gossypium hirsutum; peanut, Arachis hypogaea; maize, Zea mays). Under the conditions of plant materials versus corresponding controls, both stages of both species could find their corresponding host plants. However, H. assulta females and larvae exhibited a supersensitive and an insensitive response, respectively. Under the conditions of tobacco paired with each plant species, H. assulta females exhibited more specialized ovipositional response to tobacco than its sibling. When each plant species were combined with tobacco and tested against tobacco reference, peanut played an opposite role in the two species in their ovipositional responses to tobacco, and cotton can enhance the approaching response of H. armigera larvae when combined with tobacco. It seems that two attractive host plants also can act antagonistically with respect to host selection of the generalist via volatile exchange. Tomato should better be excluded from host list of H. assulta. PMID:28182679

  16. Cytochrome P450 and actin genes expressed in Helicoverpa zea and Helicoverpa armigera: paralogy/orthology identification, gene conversion and evolution.

    PubMed

    Li, Xianchun; Berenbaum, May R; Schuler, Mary A

    2002-03-01

    Molecular phylogenetic analysis was conducted using conserved cytoplasmic actin and diversified cytochrome P450 (P450) sequences isolated from Helicoverpa zea and Helicoverpa armigera, two species thought to be closely related based on allozyme analyses. These sequences were compared in turn with published sequences from other insects to gain insight into how different gene families evolve. In Bombyx mori and these Helicoverpa species, cytoplasmic actin genes are present as a pair of tandemly duplicated paralogs with coding sequence identities as high as 95.5% (B. mori), 98.9% (H. zea) and 98.5% (H. armigera) due to recent 5'-polar gene conversions. Phylogeny and interspecies comparisons assign the six actin genes into two orthologous groups: HaA3a/HzA3a/BmA3 and HaA3b/HzA3b/BmA4, which exhibit more similarities between H. zea and H. armigera than between Helicoverpa species and B. mori. Like the actin genes in H. zea, four CYP6B genes exist as two pairs of duplicated paralogs with recent 5'-polar gene conversions. Interspecific comparisons and phylogeny analysis identified three groups of orthologous CYP6B genes: H. zea CYP6B8 or CYP6B28/H. armigera CYP6B7, H. zea CYP6B27/H. armigera CYP6B6, and H. zea CYP6B9/H. armigera CYP6B2/Heliothis virescens CYP6B10. The low degree of divergence in the first two of these groups is comparable to allelic variation within a single species. These orthologous relationships and the high degrees of similarity in both actin and P450 genes strongly indicate that these Helicoverpa species are extremely closely related.

  17. Bitter gourd proteinase inhibitors: potential growth inhibitors of Helicoverpa armigera and Spodoptera litura.

    PubMed

    Telang, Manasi; Srinivasan, Ajay; Patankar, Aparna; Harsulkar, Abhay; Joshi, Vijay; Damle, Archana; Deshpande, Vasanti; Sainani, Mohini; Ranjekar, Prabhakar; Gupta, Gorakh; Birah, Ajanta; Rani, Seema; Kachole, Manavendra; Giri, Ashok; Gupta, Vidya

    2003-07-01

    Proteinase inhibitors (PIs) from the seeds of bitter gourd (Momordica charantia L.) were identified as strong inhibitors of Helicoverpa armigera gut proteinases (HGP). Biochemical investigations showed that bitter gourd PIs (BGPIs) inhibited more than 80% HGP activity. Electrophoretic analysis revealed the presence of two major proteins (BGPI-1 and-2) and two minor proteins (BGPI-3 and-4) having inhibitory activity against both trypsin and HGP. The major isoforms BGPI-1 and BGPI-2 have molecular mass of 3.5 and 3.0 kDa, respectively. BGPIs inhibited HGP activity of larvae fed on different host plants, on artificial diet with or without added PIs and proteinases excreted in fecal matter. Degradation of BGPI-1 by HGP showed direct correlation with accumulation of BGPI-2-like peptide, which remained stable and active against high concentrations of HGP up to 3 h. Chemical inhibitors of serine proteinases offered partial protection to BGPI-1 from degradation by HGP, suggesting that trypsin and chymotrypsin like proteinases are involved in degradation of BGPI-1. In larval feeding studies, BGPIs were found to retard growth and development of two lepidopteran pests namely Helicoverpa armigera and Spodoptera litura. This is the first report showing that BGPIs mediated inhibition of insect gut proteinases directly affects fertility and fecundity of both H. armigera and S. litura. The results advocate use of BGPIs to introduce insect resistance in otherwise susceptible plants.

  18. Data of in vitro synthesized dsRNAs on growth and development of Helicoverpa armigera

    PubMed Central

    Chikate, Yojana R.; Dawkar, Vishal V.; Barbole, Ranjit S.; Tilak, Priyadarshini V.; Gupta, Vidya S.; Giri, Ashok P.

    2016-01-01

    The data presented in this article is related to the research article “RNAi of selected candidate genes interrupts growth and development of Helicoverpa armigera” (Chikate et al., 2016) [1]. RNA interference (RNAi) is emerging as a potent insect pest control strategy over current methods and their resistance by pest. In this study we tested 15 different in vitro synthesized dsRNAs for gene silencing in Helicoverpa armigera. These dsRNAs were specific against H. armigera enzymes/proteins such as proteases like trypsins (HaTry2, 3, 4 and 6), chymotrypsin (HaChy4) and cysteine proteases such as cathepsin (HaCATHL); glutathione S-transferases (HaGST1a, 6 and 8); esterases (HaAce4, HaJHE); catalase (HaCAT); super-oxide-dismutase (HaCu/ZnSOD); fatty acid binding protein (HaFabp) and chitin deacetylase (HaCda5b). These dsRNAs were fed to second instar larvae at an optimized dose (60 µg/day) for 3 days separately. Effects of dsRNA feeding were observed in terms of larval mass gain, percentage mortality and phenotypic abnormalities in later developmental stages of H. armigera. These findings might provide potential new candidates for designing sequence-specific dsRNA as pesticide in crop protection. PMID:27222861

  19. Combining Tpi and CO1 genetic markers to discriminate invasive Helicoverpa armigera from local Helicoverpa zea (Lepidoptera:Noctuidae) populations in the southeastern United States

    USDA-ARS?s Scientific Manuscript database

    The recent establishment of the Old World pest Helicoverpa armigera into South America has had significant economic consequences and places the rest of the hemisphere at risk, emphasizing the need for improved methods of monitoring. A major complication is that a sibling species endemic to the New W...

  20. Mitochondrial genome of the cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) and comparison with other Lepidopterans.

    PubMed

    Yin, Jiao; Hong, Gui-Yun; Wang, Ai-Min; Cao, Ya-Zhong; Wei, Zhao-Jun

    2010-10-01

    We present the complete sequence of the mitochondrial genome (mitogenome) of the cotton bollworm Helicoverpa armigera. The 15,347-bp mitogenome of H. armigera was arranged in the same order described for all other sequenced lepidopterans, which differs from the most common type found in insects, due to the movement of trnM to a position 5'-upstream of trnI. The gene overlap in the H. armigera mitogenome is totally 23 bp in six locations. The H. armigera mitogenome has a total of 175 bp of intergenic spacer sequences spread over 14 regions ranging in size from 1 to 45 bp. The nucleotide composition of the whole mitogenome of H. armigera is highly A+T biased, accounting for 80.97%, with a slightly positive AT skewness and negative GC skewness, indicating the occurrence of more A than T, C more than G. The protein-encoding genes have typical mitochondrial start codons, except for cox1, which contains the unusual CGA. The cox1, cox2, and nad4 genes have incomplete stop codons (T). The lrRNA and srRNA genes are 1395 and 794-bp long, respectively. All tRNAs have a typical cloverleaf structure of mitochondrial tRNAs, except for trnS1(AGN), the dihydrouridine arm of which could not form a stable stem-loop structure. The H. armigera A+T-rich region contains a conserved structure combining the motif ATAGA and a 19-bp poly-T stretch, but absence of the 9-bp poly-A element upstream of trnM.

  1. Combining Tpi and CO1 Genetic Markers to Discriminate Invasive Helicoverpa armigera From Local Helicoverpa zea (Lepidoptera: Noctuidae) Populations in the Southeastern United States.

    PubMed

    Nagoshi, Rodney N; Gilligan, Todd M; Brambila, Julieta

    2016-10-01

    The recent establishment of the Old World pest Helicoverpa armigera (Hübner) into South America has had significant economic consequences and places the rest of the hemisphere at risk, emphasizing the need for improved methods of monitoring. A major complication is that a sibling species endemic to the New World, Helicoverpa zea (Boddie), is morphologically very similar, with the two species capable of producing fertile hybrids in the laboratory. The consequences of such hybridization in the field are uncertain, but could result in significant and unpredictable changes in the timing, range, and pesticide susceptibilities of Helicoverpa infestations. The objective here is to provide new genetic resources applicable to Helicoverpa populations in northern Florida and neighboring states (a region at risk for H. armigera) that can distinguish the two species and possible hybrids. The genetic variability in segments of the mitochondrial cytochrome oxidase 1 (CO1) and the Z-linked triosephosphate isomerase (Tpi) genes were determined for H. zea from the southeastern United States. These were compared to DNA sequences from H. armigera specimens from Morocco, Australia, and Europe. Phylogenetic network analysis showed a clear demarcation between the two species for all gene segments. These results extend earlier studies establishing CO1 as marker for discriminating the Helicoverpa species complex and introduce a new sex-linked genomic marker. The CO1 and Tpi markers in combination provide a more accurate and sensitive method than existing techniques for identifying hybridization between H. zea and H. armigera and could potentially be used to extrapolate the likely source of invasive H. armigera populations. Published by Oxford University Press on behalf of Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the US.

  2. Innate preference and learning of colour in the male cotton bollworm moth, Helicoverpa armigera.

    PubMed

    Satoh, Aya; Kinoshita, Michiyo; Arikawa, Kentaro

    2016-12-15

    We investigated colour discrimination and learning in adult males of the nocturnal cotton bollworm moth, Helicoverpa armigera, under a dim light condition. The naive moths preferred blue and discriminated the innately preferred blue from several shades of grey, indicating that the moths have colour vision. After being trained for 2 days to take nectar at a yellow disc, an innately non-preferred colour, moths learned to select yellow over blue. The choice distribution between yellow and blue changed significantly from that of naive moths. However, the dual-choice distribution of the trained moths was not significantly biased to yellow: the preference for blue is robust. We also tried to train moths to grey, which was not successful. The limited ability to learn colours suggests that H armigera may not strongly rely on colours when searching for flowers in the field, although they have the basic property of colour vision. © 2016. Published by The Company of Biologists Ltd.

  3. Impact of differential feeding on the growth and development of Helicoverpa armigera (Hubner).

    PubMed

    Rao, K Prabhakara; Radhakrishnaiah, K; Sudhakar, K

    2008-11-01

    The growth and development of Helicoverpa armigera was observed in the laboratory by feeding them on different foods viz; leaf, flower and fruit of lady finger, cotton, pigeon pea and chick pea. Based on the food ingested, food digested and food excreted as well as on weight and size of the larvae and the duration of larval period, the fruit of chick pea was found to be the most suitable food for H. armigera development, as the food ingesta and food digesta of the larvae on pigeon pea were more than on the other plants. On the pigeon pea pod the larval growth and development was fast and larval duration was short. Next to the fruit, the larvae preferred the leaf of lady finger and cotton and flower of pigeon pea. The results suggested that the larval growth and development was dependent on the feed i.e. both on the part and the type of the plant.

  4. Biochemical characterization of the cotton bollworm Helicoverpa armigera resistance to pyrethroids in Burkina Faso.

    PubMed

    Omer, S A H; Konate, G; Traore, Oumar; Traore, Ouola; Menozzi, P

    2009-07-01

    The bollworm Helicoverpa armigera is cotton plant main pest in most parts of the world. The mechanisms of the resistance of the bollworm to the pyrethroid deltamethrin were studied by comparing field strains to the reference and susceptible strain (BK77). Resistance to deltamethrin was studied using bio-assays. Results showed that the field collected strains had susceptibility 11 to 43 fold lower than that of the susceptible standard strain BK77. Activities of two types of enzymes i.e., oxidases and Glutathione-S-Transferases (GST) were significantly higher in field strains, whereas esterase activities were lower compared to that of standard strain. The increase of oxidases and GST activities and the decrease of esterase activity are at least in part, responsible for the development of resistance of H. armigera to pyrethroids.

  5. Monitoring and management strategy for Helicoverpa armigera resistance to Bt cotton in China.

    PubMed

    Wu, Kongming

    2007-07-01

    The cotton bollworm, Helicoverpa armigera, is one of the most important insect pests in cotton growing regions of China. Transgenic cotton that expresses a gene derived from the bacterium Bacillus thuringiensis (Bt) has been deployed for combating cotton bollworm since 1997. Natural refugees derived from the mixed planting system consisting of cotton, corn, soybean, vegetables, peanut and others on single-family farms of a small scale were used for delaying the evolution of resistance to Bt cotton. Susceptibility of H. armigera field populations to the Bt insecticidal protein Cry1Ac was monitored from 1997 to 2006. The results indicate that the field populations are still susceptible to Cry1Ac, and monitoring indication no apparent shifts in susceptibility in field populations of this important pest.

  6. New Resistance Mechanism in Helicoverpa armigera Threatens Transgenic Crops Expressing Bacillus thuringiensis Cry1Ac Toxin

    PubMed Central

    Gunning, Robin V.; Dang, Ho T.; Kemp, Fred C.; Nicholson, Ian C.; Moores, Graham D.

    2005-01-01

    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. PMID:15870346

  7. New resistance mechanism in Helicoverpa armigera threatens transgenic crops expressing Bacillus thuringiensis Cry1Ac toxin.

    PubMed

    Gunning, Robin V; Dang, Ho T; Kemp, Fred C; Nicholson, Ian C; Moores, Graham D

    2005-05-01

    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.

  8. Rab3 is involved in cellular immune responses of the cotton bollworm, Helicoverpa armigera.

    PubMed

    Li, Jie; Song, Cai-Xia; Li, Yu-Ping; Li, Li; Wei, Xiu-Hong; Wang, Jia-Lin; Liu, Xu-Sheng

    2015-06-01

    Rab3, a member of the Rab GTPase family, has been found to be involved in innate immunity. However, the precise function of this GTPase in innate immunity remains unknown. In this study, we identified a Rab3 gene (Ha-Rab3) from the cotton bollworm, Helicoverpa armigera and studied its roles in innate immune responses. Expression of Ha-Rab3 was upregulated in the hemocytes of H. armigera larvae after the injection of Escherichia coli or chromatography beads. The dsRNA-mediated knockdown of Ha-Rab3 gene in H. armigera larval hemocytes led to significant reduction in the phagocytosis and nodulation activities of hemocytes against E. coli, significant increase in the bacterial load in larval hemolymph, and significant reduction in the encapsulation activities of hemocytes toward invading chromatography beads. Furthermore, Ha-Rab3 knockdown significantly suppressed spreading of plasmatocytes. These results suggest that Ha-Rab3 plays important roles in H. armigera cellular immune responses, possibly by mediating spreading of hemocytes.

  9. Characterisation and inhibition studies of Helicoverpa armigera (Hübner) gut α-amylase.

    PubMed

    Kaur, Rimaljeet; Gupta, Anil K; Taggar, Gaurav K

    2015-09-01

    The survival of a devastating pest, Helicoverpa armigera, is mainly dependent on the availability of α-amylase. Therefore, characterising H. armigera α-amylase and targeting it with effective inhibitors could aid in reducing its damaging effects. H. armigera gut possessed four isozymes of α-amylase. The molecular weight of the major purified isozyme ranged from 79 to 81 kDa. The purified enzyme was identified to be α-amylase on the basis of products formed from starch. The optimum pH and temperature were 10.0 and 50 °C respectively. The activation energy was 5.7 kcal mol(-1) . The enzyme showed high activity with starch and amylopectin, whereas dextrins were poor substrates. The Michaelis constant Km with starch, amylose and amylopectin was 0.45, 1.23 and 0.11 mg mL(-1) respectively. ZnSO4 , FeSO4 , CuSO4 , citric acid, oxalic acid and salicylic acid were potent inhibitors. ZnSO4 , salicylic acid and pigeonpea α-amylase inhibitor (∼21.0 kDa) acted primarily as competitive inhibitors, FeSO4 and citric acid displayed mainly anticompetitive behaviour, while CuSO4 and oxalic acid behaved mainly as non-competitive inhibitors. The identification of effective ecofriendly inhibitors could help in managing H. armigera infestation. © 2014 Society of Chemical Industry. © 2014 Society of Chemical Industry.

  10. Genome Sequence of Acinetobacter sp. Strain HA, Isolated from the Gut of the Polyphagous Insect Pest Helicoverpa armigera

    PubMed Central

    Malhotra, Jaya; Dua, Ankita; Saxena, Anjali; Sangwan, Naseer; Mukherjee, Udita; Pandey, Neeti; Rajagopal, Raman; Khurana, Paramjit; Khurana, Jitendra P.

    2012-01-01

    In this study, Acinetobacter sp. strain HA was isolated from the midgut of a fifth-instar larva of Helicoverpa armigera. Here, we report the draft genome sequence (3,125,085 bp) of this strain that consists of 102 contigs, 2,911 predicted coding sequences, and a G+C content of 41%. PMID:22933775

  11. Relationships of Helicoverpa armigera, Ostrinia nubilalis and Fusarium verticillioides on MON 810 Maize

    PubMed Central

    Darvas, Béla; Bánáti, Hajnalka; Takács, Eszter; Lauber, Éva; Szécsi, Árpád; Székács, András

    2011-01-01

    MON 810 maize was developed against Ostrinia nubilalis and is suggested to indirectly decrease Fusarium spp. infestation in maize ears. To evaluate this effect, co-occurrence of insect and fungal pests on MON 810 maize was studied. During 2009, exceptionally high maize ear infestation occurred in Julianna-major (Hungary). From investigation of some thousands of maize ears, the majority of the larval damage originated from Helicoverpa armigera larvae, while O. nubilalis larvae contributed significant damage only at a single plot. Fusarium verticillioides infection appeared only in a small portion (∼20–30%) of the insect damaged cobs. H. armigera and O. nubilalis larvae feeding on F. verticillioides mycelia can distribute its conidia with their fecal pellets. MON 810 maize showed 100% efficacy against O. nubilalis in the stem, but lower efficacy against O. nubilalis and H. armigera in maize ears. The ∼Cry1Ab toxin content of maize silk, the entry site of H. armigera, was lower than that in the leaves/stem/husk leaves of MON 810. Fusarium-infected MON 810 cobs are rarely found and only after larval damage by O. nubilalis. H. armigera larvae could not tolerate well F. verticillioides infected food and attempted to move out from the infected cobs. For further feeding they re-entered the maize ears through the 8–12 husk leaves, but in the case of the MON 810 variety, they usually could not reach the kernels. Apical damage on cobs resulted in only a minor (about one-tenth of the cob) decrease in yield. PMID:26467495

  12. The Potential Distribution of Invading Helicoverpa armigera in North America: Is It Just a Matter of Time?

    PubMed Central

    Kriticos, Darren J.; Ota, Noboru; Hutchison, William D.; Beddow, Jason; Walsh, Tom; Tay, Wee Tek; Borchert, Daniel M.; Paula-Moreas, Silvana V.; Czepak, Cecília; Zalucki, Myron P.

    2015-01-01

    Helicoverpa armigera has recently invaded South and Central America, and appears to be spreading rapidly. We update a previously developed potential distribution model to highlight the global invasion threat, with emphasis on the risks to the United States. The continued range expansion of H. armigera in Central America is likely to change the invasion threat it poses to North America qualitatively, making natural dispersal from either the Caribbean islands or Mexico feasible. To characterise the threat posed by H. armigera, we collated the value of the major host crops in the United States growing within its modelled potential range, including that area where it could expand its range during favourable seasons. We found that the annual value of crops that would be exposed to H. armigera totalled approximately US$78 billion p.a., with US$843 million p.a. worth growing in climates that are optimal for the pest. Elsewhere, H. armigera has developed broad-spectrum pesticide resistance; meaning that if it invades the United States, protecting these crops from significant production impacts could be challenging. It may be cost-effective to undertake pre-emptive biosecurity activities such as slowing the spread of H. armigera throughout the Americas, improving the system for detecting H. armigera, and methods for rapid identification, especially distinguishing between H. armigera, H. zea and potential H. armigera x H. zea hybrids. Developing biological control programs, especially using inundative techniques with entomopathogens and parasitoids could slow the spread of H. armigera, and reduce selective pressure for pesticide resistance. The rapid spread of H. armigera through South America into Central America suggests that its spread into North America is a matter of time. The likely natural dispersal routes preclude aggressive incursion responses, emphasizing the value of preparatory communication with agricultural producers in areas suitable for invasion by H

  13. The potential distribution of invading Helicoverpa armigera in North America: is it just a matter of time?

    PubMed

    Kriticos, Darren J; Ota, Noboru; Hutchison, William D; Beddow, Jason; Walsh, Tom; Tay, Wee Tek; Borchert, Daniel M; Paula-Moraes, Silvana V; Paula-Moreas, Silvana V; Czepak, Cecília; Zalucki, Myron P

    2015-01-01

    Helicoverpa armigera has recently invaded South and Central America, and appears to be spreading rapidly. We update a previously developed potential distribution model to highlight the global invasion threat, with emphasis on the risks to the United States. The continued range expansion of H. armigera in Central America is likely to change the invasion threat it poses to North America qualitatively, making natural dispersal from either the Caribbean islands or Mexico feasible. To characterise the threat posed by H. armigera, we collated the value of the major host crops in the United States growing within its modelled potential range, including that area where it could expand its range during favourable seasons. We found that the annual value of crops that would be exposed to H. armigera totalled approximately US$78 billion p.a., with US$843 million p.a. worth growing in climates that are optimal for the pest. Elsewhere, H. armigera has developed broad-spectrum pesticide resistance; meaning that if it invades the United States, protecting these crops from significant production impacts could be challenging. It may be cost-effective to undertake pre-emptive biosecurity activities such as slowing the spread of H. armigera throughout the Americas, improving the system for detecting H. armigera, and methods for rapid identification, especially distinguishing between H. armigera, H. zea and potential H. armigera x H. zea hybrids. Developing biological control programs, especially using inundative techniques with entomopathogens and parasitoids could slow the spread of H. armigera, and reduce selective pressure for pesticide resistance. The rapid spread of H. armigera through South America into Central America suggests that its spread into North America is a matter of time. The likely natural dispersal routes preclude aggressive incursion responses, emphasizing the value of preparatory communication with agricultural producers in areas suitable for invasion by H

  14. A perspective on management of Helicoverpa armigera: transgenic Bt cotton, IPM, and landscapes.

    PubMed

    Downes, Sharon; Kriticos, Darren; Parry, Hazel; Paull, Cate; Schellhorn, Nancy; Zalucki, Myron P

    2017-03-01

    Helicoverpa armigera is a major pest of agriculture, horticulture and floriculture throughout the Old World and recently invaded parts of the New World. We overview of the evolution in thinking about the application of area-wide approaches to assist with its control by the Australian Cotton Industry to highlight important lessons and future challenges to achieving the same in the New World. An over-reliance of broad-spectrum insecticides led to Helicoverpa spp. in Australian cotton rapidly became resistant to DDT, synthetic pyrethroids, organophosphates, carbamates and endosulfan. Voluntary strategies were developed to slow the development of insecticide resistance, which included rotating chemistries and basing spray decisions on thresholds. Despite adoption of these practices, insecticide resistance continued to develop until the introduction of genetically modified cotton provided a platform for augmenting Integrated Pest Management in the Australian cotton industry. Compliance with mandatory resistance management plans for Bt cotton necessitated a shift from pest control at the level of individual fields or farms towards a coordinated area-wide landscape approach. Our take-home message for control of H. armigera is that resistance management is essential in genetically modified crops and must be season long and area-wide to be effective. © 2016 Society of Chemical Industry.

  15. Expressional divergence of the fatty acid-amino acid conjugate-hydrolyzing aminoacylase 1 (L-ACY-1) in Helicoverpa armigera and Helicoverpa assulta.

    PubMed

    Cheng, Qian; Gu, Shaohua; Liu, Zewen; Wang, Chen-Zhu; Li, Xianchun

    2017-08-18

    How FACs-producing generalist and specialist herbivores regulate their FACs-hydrolyzing enzyme L-ACY-1 to balance FACs' beneficial vs. detrimental effects remains unknown. To address this question, we compared L-ACY-1 expression in Helicoverpa armigera and Helicoverpa assulta, a pair of closely related sibling species differing mainly in their host range, by the same sets of hostplants, protein to digestible carbohydrate (P:C) ratios, or allelochemical. L-ACY-1 expression remained low/unchanged in H. armigera, but was induced by hot pepper fruits and repressed by cotton bolls in H. assulta. The representative allelochemicals of the tested hostplants significantly (capsaicin) or insignificantly (gossypol and nicotine) induced L-ACY-1 expression in H. armigera, but insignificantly inhibited (capsaicin and gossypol) or induced (nicotine) it in H. assulta. L-ACY-1 expression remained low/unaltered on balanced (P50:C50 and P53:C47) or protein-biased diets and induced on carbohydrate-biased diets in H. armigera, but was at the highest level on balanced diets and reduced on either protein- or carbohydrate-biased diets in H. assulta. Furthermore, L-ACY-1 expression was significantly higher in H. assulta than in H. armigera for most of feeding treatments. Such expressional divergences suggest that FACs are utilized mainly for removal of excessive nitrogen in generalists but for nitrogen assimilation in specialists.

  16. Characterization of the metabolic transformation of thiamethoxam to clothianidin in Helicoverpa armigera larvae by SPE combined UPLC-MS/MS and its relationship with the toxicity of thiamethoxam to Helicoverpa armigera larvae.

    PubMed

    Fan, Yinjun; Shi, Xueyan

    2017-09-01

    In order to characterize the metabolic transformation of thiamethoxam (TMX) to clothianidin (CLO) in Helicoverpa armigera larvae and clarify its relationship with the insecticidal toxicity of TMX, method for determination of TMX and its metabolite clothianidin (CLO) residues in H. armigera larvae by solid phase extraction (SPE) combined UPLC-MS/MS was established. Following acetonitrile extraction and purification by SPE on florisil cartridge and C18 cartridge sequently, and cleanup by PSA adsorption, TMX and CLO residues in H. armigera larvae were successfully determined by UPLC-MS/MS. By using the established method, the concentration-time curves of TMX and its metabolite CLO in H. armigera larvae in vivo and metabolism of TMX by microsome of H. armigera larvae midguts in vitro were studied. TMX was quickly eliminated from H. armigera larvae with the elimination half-life as 4.2h. Meanwhile, only a small amount of CLO was formed from TMX metabolism, with the maximum CLO level in H. armigera larvae only accounts for the metabolic transformation of 7.99% of TMX, at 10h after intravenous TMX administration. Our results suggested that the low insecticidal efficacy of TMX against H. armigera larvae was related with the rapidly elimination of TMX from H. armigera larvae, meanwhile, CLO as TMX metabolite at a very low level in vivo didn't contribute to TMX toxicity to H. armigera larvae. In H. armigera larvae, TMX didn't act as proinsecticide for CLO in insecticidal efficacy of TMX. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Nutritional performance of the tomato fruit borer, Helicoverpa armigera, on different tomato cultivars.

    PubMed

    Kouhi, Davoud; Naseri, Bahram; Golizadeh, Ali

    2014-01-01

    The development and cultivation of tomato cultivars that are resistant to the tomato fruit borer, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), are very limited in Iran and other parts of the world because of the lack of information about resistant tomato cultivars to minimize the use of insecticides. Therefore, the present study was carried out to identify alternative methods to chemical control. Nutritional performance of the larval stages (fourth, fifth, and sixth instars) of H. armigera on fruit of eight tomato cultivars, including SUN 6108 f1, Rio grande UG, Korral, Super strain B, CH falat, Hed rio grande, Cal.JN3, and Super crystal, was studied under laboratory conditions. Fourth instars reared on CH falat and SUN 6108 f1 respectively showed the highest and lowest values of approximate digestibility. The highest values of efficiency of conversion of ingested food and efficiency of conversion of digested food of fifth instars were on Super strain B. The relative consumption rate and relative growth rate values of the sixth instars were the highest on Korral. The highest and lowest values of consumption index of sixth instars were on Super strain B and Hed rio grande, respectively. The efficiency of conversion of ingested food and efficiency of conversion of digested food values of whole larval instars were the highest on Hed rio grande and lowest on Rio grande UG. The results of nutritional indices indicated that Rio grande UG is an unsuitable host for H. armigera.

  18. Acute, Sublethal, and Combination Effects of Azadirachtin and Bacillus thuringiensis on the Cotton Bollworm, Helicoverpa armigera

    PubMed Central

    Abedi, Zahra; Saber, Moosa; Vojoudi, Samad; Mahdavi, Vahid; Parsaeyan, Ehsan

    2014-01-01

    The cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a polyphagous and cosmopolitan insect pest that causes damage to various plants. In this study, the lethal and sublethal effects of azadirachtin and Bacillus thuringiensis Berliner sub sp. kurstaki (Bacillales: Bacillaceae) were evaluated on third instar H. armigera under laboratory conditions. The LC50 values of azadirachtin and Bt were 12.95 and 96.8 µg a.i./mL, respectively. A total mortality of 56.7% was caused on third instar larvae when LC20 values of the insecticides were applied in combination with each other. The LT50 values of azadirachtin and Bt were 4.8 and 3.6 days, respectively. The results of the sublethal study showed that the application of LC30 value of azadirachtin and Bt reduced the larval and pupal weight and increased larval and pupal duration of H. armigera. The longevity and fecundity of female adults were affected significantly by the insecticides. Female fecundity was reduced by the treatments, respectively. The lowest adult emergence ratio and pupation ratio were observed in the azadirachtin treatment. The results indicated that both insecticides have high potential for controlling of the pest. PMID:25373177

  19. Differential antibiosis against Helicoverpa armigera exerted by distinct inhibitory repeat domains of Capsicum annuum proteinase inhibitors.

    PubMed

    Joshi, Rakesh S; Gupta, Vidya S; Giri, Ashok P

    2014-05-01

    Plant defensive serine proteinase inhibitors (PIs) are known to have negative impact on digestive physiology of herbivore insects and thus have a crucial role in plant protection. Here, we have assessed the efficacy and specificity of three previously characterized inhibitory repeat domain (IRD) variants from Capsicum annuum PIs viz., IRD-7, -9 and -12 against gut proteinases from Helicoverpa armigera. Comparative study of in silico binding energy revealed that IRD-9 possesses higher affinity towards H. armigera serine proteinases as compared to IRD-7 and -12. H. armigera fed on artificial diet containing 5 TIU/g of recombinant IRD proteins exhibited differential effects on larval growth, survival rate and other nutritional parameters. Major digestive gut trypsin and chymotrypsin genes were down regulated in the IRD fed larvae, while few of them were up-regulated, this indicate alterations in insect digestive physiology. The results corroborated with proteinase activity assays and zymography. These findings suggest that the sequence variations among PIs reflect in their efficacy against proteinases in vitro and in vivo, which also could be used for developing tailor-made multi-domain inhibitor gene(s).

  20. Bacterial Expression and Kinetic Analysis of Carboxylesterase 001D from Helicoverpa armigera

    PubMed Central

    Li, Yongqiang; Liu, Jianwei; Lu, Mei; Ma, Zhiqing; Cai, Chongling; Wang, Yonghong; Zhang, Xing

    2016-01-01

    Carboxylesterasesare an important class of detoxification enzymes involved in insecticide resistance in insects. A subgroup of Helicoverpa armigera esterases, known as Clade 001, was implicated in organophosphate and pyrethroid insecticide resistance due to their overabundance in resistant strains. In this work, a novel carboxylesterasegene 001D of H. armigera from China was cloned, which has an open reading frame of 1665 nucleotides encoding 554 amino acid residues. We used a series of fusion proteins to successfully express carboxylesterase 001D in Escherichia coli. Three different fusion proteins were generated and tested. The enzyme kinetic assay towards 1-naphthyl acetate showed all three purified fusion proteins are active with a Kcat between 0.35 and 2.29 s−1, and a Km between 7.61 and 19.72 μM. The HPLC assay showed all three purified fusion proteins had low but measurable hydrolase activity towards β-cypermethrin and fenvalerate insecticides (specific activities ranging from 0.13 to 0.67 μM·min−1·(μM−1·protein)). The enzyme was stable up to 40 °C and at pH 6.0–11.0. The results imply that carboxylesterase 001D is involved in detoxification, and this moderate insecticide hydrolysis may suggest that overexpression of the gene to enhance insecticide sequestration is necessary to allow carboxylesterases to confer resistance to these insecticides in H. armigera. PMID:27049381

  1. Insecticidal Potential of Defense Metabolites from Ocimum kilimandscharicum against Helicoverpa armigera

    PubMed Central

    Thulasiram, Hirekodathakallu V.; Kulkarni, Mahesh J.; Giri, Ashok P.

    2014-01-01

    Genus Ocimum contains a reservoir of diverse secondary metabolites, which are known for their defense and medicinal value. However, the defense-related metabolites from this genus have not been studied in depth. To gain deeper insight into inducible defense metabolites, we examined the overall biochemical and metabolic changes in Ocimum kilimandscharicum that occurred in response to the feeding of Helicoverpa armigera larvae. Metabolic analysis revealed that the primary and secondary metabolism of local and systemic tissues in O. kilimandscharicum was severely affected following larval infestation. Moreover, levels of specific secondary metabolites like camphor, limonene and β-caryophyllene (known to be involved in defense) significantly increased in leaves upon insect attack. Choice assays conducted by exposing H. armigera larvae on O. kilimandscharicum and tomato leaves, demonstrated that O. kilimandscharicum significantly deters larval feeding. Further, when larvae were fed on O. kilimandscharicum leaves, average body weight decreased and mortality of the larvae increased. Larvae fed on artificial diet supplemented with O. kilimandscharicum leaf extract, camphor, limonene and β-caryophyllene showed growth retardation, increased mortality rates and pupal deformities. Digestive enzymes of H. armigera - namely, amylase, protease and lipase- showed variable patterns after feeding on O. kilimandscharicum, which implies striving of the larvae to attain required nutrition for growth, development and metamorphosis. Evidently, selected metabolites from O. kilimandscharicum possess significant insecticidal activity. PMID:25098951

  2. Developmental inhibition and DNA damage of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) by gamma radiation.

    PubMed

    Kim, Junheon; Chung, Soon-Oh; Jang, Miyeon; Jang, Sin Ae; Park, Chung Gyoo

    2015-01-01

    Gamma radiation on Helicoverpa armigera Hübner was performed to assess developmental inhibition and to identify a potential quarantine treatment dose of the radiation. Gamma radiation ((60)Co) treatment at different doses of 50, 100, 200, 300, and 400 Gy was carried out with egg, larvae, pupae and adults of H. armigera. Gamma radiation induced developmental inhibition of all stages of H. armigera. The effective dose values required for inhibition 99% (ED(99)) of hatching, pupation and adult emergence from the irradiated eggs were 550.7, 324.9 and 136.4 Gy, respectively. ED(99) values for inhibition of the larvae to adult emergence was 200.0 Gy. Irradiation on pupae could not completely inhibit adult emergence even at 400 Gy. ED(99) value for inhibition of F(1) egg hatchability from the irradiated adults was estimated to be 229.5 Gy. This study suggests that gamma radiation is a possible alternative to phytosanitary treatments. Irradiation treatment with minimum dose of 200 Gy can be suggested as optimum dose for larval treatment in quarantine.

  3. Successes and challenges of managing resistance in Helicoverpa armigera to Bt cotton in Australia.

    PubMed

    Downes, Sharon; Mahon, Rod

    2012-01-01

    Bt cotton has been gradually released and adopted by Australian growers since 1996. It was initially deployed in Australia primarily to control the polyphagous pest Helicoverpa armigera (Hübner), which in the 1990s became increasingly difficult to control due to widespread resistance to synthetic chemical insecticides. Bt-cotton has become a key tool in a program of integrated pest management for the production system that reduces pesticide dependence and the problems associated with its use. Herein we overview the deployment of Bt cotton in Australia including its performance and the approaches used to prolong the evolution of resistance to it by H. armigera. An integral component of this approach is monitoring resistance in this pest. We outline resistance screening methods, as well as the characteristics of resistant strains of H. armigera that have been isolated from field populations, or selected in the laboratory. We then highlight the successes and challenges for Bt cotton in Australia by way of discussing adaptive resistance management in light of potential changes in resistance.

  4. Insecticidal activity of a novel fatty acid amide derivative from Streptomyces species against Helicoverpa armigera.

    PubMed

    Gopalakrishnan, Subramaniam; Rajendran, Vijayabharathi; Arumugam, Sathya; Sharma, Hari C; Vadlamudi, Srinivas; Bhimineni, Ratna Kumari; V Gonzalez, Susana; M Melø, Torunn; Simic, Nebojsa

    2016-03-08

    Helicoverpa armigera, an important pest causes serious damage to grain legumes. The main objective of this study was to isolate and identify the metabolite against H. armigera from a previously characterised Streptomyces sp. CAI-155. The culture filtrate of CAI-155 was extracted using Diaion HP-20 and the active fractions were fractionated on Silica and C18 column chromatography. The C18 active fraction was further fractionated on Silica gel 60 F254 thin layer chromatography (TLC). The most active fraction (Rf 0.64) purified from TLC led to the identification of a novel metabolite N-(1-(2,2-dimethyl-5-undecyl-1,3-dioxolan-4-yl)-2-hydroxyethyl)stearamide by spectral studies. The purified metabolite showed 70-78% mortality in 2nd instar H. armigera by diet impregnation assay, detached leaf assay and greenhouse assay. The LD50 and LD90 values of the purified metabolite were 627 and 2276 ppm, respectively. Hence, this novel metabolite can be exploited for pest management in future.

  5. Nutritional Performance of the Tomato Fruit Borer, Helicoverpa armigera, on Different Tomato Cultivars

    PubMed Central

    Kouhi, Davoud; Naseri, Bahram; Golizadeh, Ali

    2014-01-01

    The development and cultivation of tomato cultivars that are resistant to the tomato fruit borer, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), are very limited in Iran and other parts of the world because of the lack of information about resistant tomato cultivars to minimize the use of insecticides. Therefore, the present study was carried out to identify alternative methods to chemical control. Nutritional performance of the larval stages (fourth, fifth, and sixth instars) of H. armigera on fruit of eight tomato cultivars, including SUN 6108 f1, Rio grande UG, Korral, Super strain B, CH falat, Hed rio grande, Cal.JN3, and Super crystal, was studied under laboratory conditions. Fourth instars reared on CH falat and SUN 6108 f1 respectively showed the highest and lowest values of approximate digestibility. The highest values of efficiency of conversion of ingested food and efficiency of conversion of digested food of fifth instars were on Super strain B. The relative consumption rate and relative growth rate values of the sixth instars were the highest on Korral. The highest and lowest values of consumption index of sixth instars were on Super strain B and Hed rio grande, respectively. The efficiency of conversion of ingested food and efficiency of conversion of digested food values of whole larval instars were the highest on Hed rio grande and lowest on Rio grande UG. The results of nutritional indices indicated that Rio grande UG is an unsuitable host for H. armigera. PMID:25204681

  6. X-ray radiation and development inhibition of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

    NASA Astrophysics Data System (ADS)

    Kim, Junheon; Jung, Soon-Oh; Jang, Sin Ae; Kim, Jeongmin; Park, Chung Gyoo

    2015-10-01

    Effect of X-ray radiation on the development inhibition was evaluated for all stages of the life cycle of Helicoverpa armigera to determine a radiation dose for potential quarantine treatment against the insect. ED99 values for inhibition of hatching, pupation, and adult emergence from irradiated eggs were 413, 210, and 154 Gy, respectively. ED99 values for inhibition of pupation and adult emergence from irradiated larvae were 221 and 167 Gy, respectively. Pupa was the most tolerant to X-ray radiation. ED99 value for inhibition of adult emergence from irradiated pupae was as high as 2310 Gy, whereas that for inhibition of F1 egg hatching was only 66 Gy. ED99 value for inhibition of hatching of F1 eggs which were laid by irradiated adults was estimated to 194 Gy. X-ray irradiation against H. armigera is recommended as an alternative method to methyl bromide fumigation for phytosanitary treatments during quarantine. X-ray radiation dose of 200 Gy is proposed as a potential quarantine treatment dose for H. armigera eggs and larvae.

  7. Resurgence of the cotton bollworm Helicoverpa armigera in northern Greece associated with insecticide resistance.

    PubMed

    Mironidis, George K; Kapantaidaki, Despina; Bentila, Maria; Morou, Evangelia; Savopoulou-Soultani, M; Vontas, John

    2013-08-01

    Helicoverpa armigera has been controlled effectively with chemical insecticides in the major cotton crop production areas of northern Greece for many years. However, a resurgence of the pest was observed in 2010, which significantly affected crop production. During a 4-year survey (2007-2010), we examined the insecticide resistance status of H. armigera populations from two major and representative cotton production areas in northern Greece against seven insecticides (chlorpyrifos, diazinon, methomyl, alpha-cypermethrin, cypermethrin, gamma-cyhalothrin and endosulfan). Full dose-response bioassays on third instar larvae were performed by topical application. Lethal doses at 50% were estimated by probit analysis and resistance factors (RF) were calculated, compared to a susceptible laboratory reference strain. Resistance levels were relatively moderate until 2009, with resistance ratios below 10-fold for organophosphates and carbamates and up to 16-fold for the pyrethroid alpha-cypermethrin. However, resistance rose to 46- and 81-fold for chlorpyrifos and alpha-cypermethrin, respectively in 2010, when the resurgence of the pest was observed. None of the known pyrethroid resistance mutations were found in the pyrethroid-resistant insects. The possible association between resistance and H. armigera resurgence in Greece is discussed. © 2012 Institute of Zoology, Chinese Academy of Sciences.

  8. Characterization of Three Novel SINE Families with Unusual Features in Helicoverpa armigera

    PubMed Central

    Wang, Jianjun; Wang, Aina; Han, Zhaojun; Zhang, Zan; Li, Fei; Li, Xianchun

    2012-01-01

    Although more than 120 families of short interspersed nuclear elements (SINEs) have been isolated from the eukaryotic genomes, little is known about SINEs in insects. Here, we characterize three novel SINEs from the cotton bollworm, Helicoverpa armigera. Two of them, HaSE1 and HaSE2, share similar 5′ -structure including a tRNA-related region immediately followed by conserved central domain. The 3′ -tail of HaSE1 is significantly similar to that of one LINE retrotransposon element, HaRTE1.1, in H. armigera genome. The 3′ -region of HaSE2 showed high identity with one mariner-like element in H. armigera. The third family, termed HaSE3, is a 5S rRNA-derived SINE and shares both body part and 3′-tail with HaSE1, thus may represent the first example of a chimera generated by recombination between 5S rRNA and tRNA-derived SINE in insect species. Further database searches revealed the presence of these SINEs in several other related insect species, but not in the silkworm, Bombyx mori, indicating a relatively narrow distribution of these SINEs in Lepidopterans. Apart from above, we found a copy of HaSE2 in the GenBank EST entry for the cotton aphid, Aphis gossypii, suggesting the occurrence of horizontal transfer. PMID:22319625

  9. Baseline Susceptibility of Helicoverpa armigera (Lepidoptera: Noctuidae) to Indoxacarb, Emamectin Benzoate, and Chlorantraniliprole in Australia.

    PubMed

    Bird, Lisa J

    2015-02-01

    Baseline susceptibility of Helicoverpa armigera (Hübner) to emamectin benzoate, chlorantraniliprole, and indoxacarb was determined in feeding assays on insecticide-incorporated artificial diet in the laboratory. The intraspecific variation of H. armigera was established from field populations collected between September 2012 and March 2013, primarily from commercial farms across eastern Australia. Emamectin benzoate had the highest toxicity with a median lethal concentration (LC50) of 0.01 µg/ml diet (n=20 strains). The LC50 for chlorantraniliprole was 0.03 µg/ml diet (n=21 strains), while indoxacarb had the lowest relative toxicity with an average LC50 of 0.3 µg/ml diet (n=22 strains). Variation in susceptibility amongst field strains was 2.3-fold for emamectin benzoate and 2.9-fold for chlorantraniliprole and indoxacarb. Discriminating concentrations of 0.2, 1, and 12 µg of insecticide per milliliter of diet for emamectin benzoate, chlorantraniliprole, and indoxacarb, respectively, were calculated from toxicological data from field H. armigera strains as a first step in resistance management of these classes of insecticide in Australia. The low intraspecific tolerance, high slope values, and goodness-of-fit to a probit binomial model obtained in this study suggest that a feeding assay using diet incorporated insecticide is an effective laboratory method for measuring the dose-responses of these classes of insecticides in H. armigera. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  10. Exploitation of wild Cicer reticulatum germplasm for resistance to Helicoverpa armigera.

    PubMed

    Sharma, H C; Pampapathy, G; Lanka, S K; Ridsdill-Smith, T J

    2005-12-01

    In the absence of high levels of resistance to Helicoverpa armigera (Hübner) in the cultivated germplasm of chickpea, we evaluated accessions of Cicer spp. mostly Cicer reticulatum Ladzinsky, for resistance to this important pest. Under multichoice conditions in the field, 10 accessions showed lower leaf damage and lower numbers of eggs, larvae, or both of H. armigera. Of these, IG 69960, IG 72934, and IG 72936 showed significantly lower leaf feeding than the cultivated genotypes or other accessions at the vegetative and reproductive stages. Larval weight was lower or comparable with that on C. bijugum (IG 70019) and C. judaicum (IG 70032) in C. reticulatum accessions IG 72933, IG 72934, IG 72936, and IG 72953 at the seedling stage and on IG 69960 and IG 72934 at the flowering stage. The accessions showing resistance to H. armigera in the field and laboratory conditions were placed in different groups, indicating the presence of diversity in C. reticulatum accessions for resistance to this pest. Less than seven larvae survived on IG 70020, IG 72940, IG 72948, and IG 72949, and IG 72964 compared with 12 on ICC 506. Larval and total developmental periods were prolonged by 6-15 and 3-8 d, respectively, on C. reticultatum accessions compared with those on ICCC 37. Less than five larvae pupated on the C. reticulatum accessions (except IG 72958 and ICC 17163) compared with 11 in ICCC 37. Accessions showing lower leaf feeding and adverse effects on the survival and development can be used in increasing the levels and diversifying the basis of resistance to H. armigera in chickpea.

  11. Biochemical, structural and functional diversity between two digestive α-amylases from Helicoverpa armigera.

    PubMed

    Bhide, Amey J; Channale, Sonal M; Patil, Sucheta S; Gupta, Vidya S; Ramasamy, Sureshkumar; Giri, Ashok P

    2015-09-01

    Helicoverpa armigera (Lepidoptera) feeds on various plants using diverse digestive enzymes as one of the survival tool-kit. The aim of the present study was to understand biochemical properties of recombinant α-amylases of H. armigera viz., HaAmy1 and HaAmy2. The open reading frames of HaAmy1 and HaAmy2 were cloned in Pichia pastoris and expressed heterologously. Purified recombinant enzymes were characterized for their biochemical and biophysical attributes using established methods. Sequence alignment and homology modeling showed that HaAmy1 and HaAmy2 were conserved in their amino acid sequences and structures. HaAmy1 and HaAmy2 showed optimum activity at 60°C; however, they differed in their optimum pH. Furthermore, HaAmy2 showed higher affinity for starch and amylopectin whereas HaAmy1 had higher catalytic efficiency. HaAmy1 and HaAmy2 were inhibited to the same magnitude by a synthetic amylase inhibitor (acarbose) while wheat amylase inhibitor showed about 2-fold higher inhibition of HaAmy1 than HaAmy2 at pH7 while 6-fold difference at pH11. Interactions of HaAmy1 and HaAmy2 with wheat amylase inhibitor revealed 2:1 stoichiometric ratio and much more complex interaction with HaAmy1. The diversity of amylases in perspective of their biochemical and biophysical properties, and their differential interactions with amylase inhibitors signify the potential role of these enzymes in adaptation of H. armigera on diverse plant diets. Characterization of digestive enzymes of H. armigera provides the molecular basis for the polyphagous nature and thus could assist in designing future strategies for the insect control. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Differential immunosuppression by Campoletis chlorideae eggs and ichnovirus in larvae of Helicoverpa armigera and Spodoptera exigua.

    PubMed

    Han, Li-Bin; Yin, Li-Hong; Huang, Ling-Qiao; Wang, Chen-Zhu

    2015-09-01

    The ichneumonid wasp, Campoletis chlorideae Uchida, successfully develops in the cotton bollworm Helicoverpa armigera (Hübner), but rarely survives in the beet armyworm Spodoptera exigua (Hübner) due to the encapsulation by host immunity. In this study, we investigated the role of C. chlorideae ichnovirus (CcIV) and eggs in the evasion of the host immune system. Washed eggs of different types, immature, mature, newly laid, or pretreated with protease K, were injected alone or with the calyx fluid containing CcIV into the larvae of H. armigera and S. exigua. In H. armigera, when injected with washed eggs alone, only 9.5% of the mature eggs were encapsulated at 24h post-injection. This is much lower than that of the immature eggs (100%), mature eggs pretreated with protease K (100%) and newly laid eggs (54.4%). No encapsulation was observed when the washed eggs were co-injected with calyx fluid at 24h post-injection. Conversely, the eggs in all treatments were encapsulated in S. exigua. Electron microscopic observations of parasitoid eggs showed structural differences between the surfaces of the mature and other kinds of eggs. The injected CcIV decreased the numbers of host hemocytes and suppressed the spreading ability of plasmatocytes and granulocytes in H. armigera, but had little effect on the hemocytes from S. exigua. In conclusion, the C. chlorideae egg provides a passive protection against encapsulation by itself, and CcIV supplies an active protection through disrupting host immune responses. These coordinated protections are host-specific, implying their role in host range determination. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. Life table and consumption capacity of corn earworm, Helicoverpa armigera, fed asparagus, Asparagus officinalis.

    PubMed

    Jha, Ratna Kumar; Tuan, Shu-Jen; Chi, Hsin; Tang, Li-Cheng

    2014-03-01

    The life table and consumption rate of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) reared on asparagus, Asparagus officinalis L. (Asparagales: Asparagaceae) were studied under laboratory conditions to assess their interaction. Development, survival, fecundity, and consumption data were analyzed by the age-stage, twosex life table. This study indicated that asparagus is a natural host of H. armigera. However, the poor nutritional content in asparagus foliage and the poor fitness of H. armigera that fed on asparagus indicated that asparagus is a suboptimal host in comparison to hybrid sweet corn. The uncertainty associated with life table parameters was estimated by using jackknife and bootstrap techniques, and the results were compared for statistical inference. The intrinsic rate of increase (r), finite rate of increase (λ), net reproductive rate (R0), and mean generation time (T) were estimated by the jackknife technique to be 0.0780 day(-1), 1.0811 day(-1), 67.4 offspring, and 54.8 days, respectively, while those estimated by the bootstrap technique were 0.0752 day(-1), 1.0781 day(-1), 68.0 offspring, and 55.3 days, respectively. The net consumption rate of H. armigera, as estimated by the jackknife and bootstrap technique, was 1183.02 and 1132.9 mg per individual, respectively. The frequency distribution of sample means obtained by the jackknife technique failed the normality test, while the bootstrap results fit the normal distribution well. By contrast, the relationship between the mean fecundity and the net reproductive rate, as estimated by the bootstrap technique, was slightly inconsistent with the relationship found by mathematical proof. The application of the jackknife and bootstrap techniques in estimating population parameters requires further examination. This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.

  14. Life Table and Consumption Capacity of Corn Earworm, Helicoverpa armigera, Fed Asparagus, Asparagus officinalis

    PubMed Central

    Jha, Ratna Kumar; Tuan, Shu-Jen; Chi, Hsin; Tang, Li-Cheng

    2014-01-01

    The life table and consumption rate of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) reared on asparagus, Asparagus officinalis L. (Asparagales: Asparagaceae) were studied under laboratory conditions to assess their interaction. Development, survival, fecundity, and consumption data were analyzed by the age-stage, two-sex life table. This study indicated that asparagus is a natural host of H. armigera. However, the poor nutritional content in asparagus foliage and the poor fitness of H. armigera that fed on asparagus indicated that asparagus is a suboptimal host in comparison to hybrid sweet corn. The uncertainty associated with life table parameters was estimated by using jackknife and bootstrap techniques, and the results were compared for statistical inference. The intrinsic rate of increase (r), finite rate of increase (λ), net reproductive rate (R0), and mean generation time (T) were estimated by the jackknife technique to be 0.0780 day-1, 1.0811 day-1, 67.4 offspring, and 54.8 days, respectively, while those estimated by the bootstrap technique were 0.0752 day-1, 1.0781 day-1, 68.0 offspring, and 55.3 days, respectively. The net consumption rate of H. armigera, as estimated by the jackknife and bootstrap technique, was 1183.02 and 1132.9 mg per individual, respectively. The frequency distribution of sample means obtained by the jackknife technique failed the normality test, while the bootstrap results fit the normal distribution well. By contrast, the relationship between the mean fecundity and the net reproductive rate, as estimated by the bootstrap technique, was slightly inconsistent with the relationship found by mathematical proof. The application of the jackknife and bootstrap techniques in estimating population parameters requires further examination. PMID:25373181

  15. Nitrate reductase and nitrite as additional components of defense system in pigeonpea (Cajanus cajan L.) against Helicoverpa armigera herbivory.

    PubMed

    Kaur, Rimaljeet; Gupta, Anil Kumar; Taggar, Gaurav Kumar

    2014-10-01

    Amylase inhibitors serve as attractive candidates of defense mechanisms against insect attack. Therefore, the impediment of Helicoverpa armigera digestion can be the effective way of controlling this pest population. Nitrite was found to be a potent mixed non-competitive competitive inhibitor of partially purified α-amylase of H. armigera gut. This observation impelled us to determine the response of nitrite and nitrate reductase (NR) towards H. armigera infestation in nine pigeonpea genotypes (four moderately resistant, three intermediate and two moderately susceptible). The significant upregulation of NR in moderately resistant genotypes after pod borer infestation suggested NR as one of the factors that determine their resistance status against insect attack. The pod borer attack caused greater reduction of nitrate and significant accumulation of nitrite in moderately resistant genotypes. The activity of nitrite reductase (NiR) was also enhanced more in moderately resistant genotypes than moderately susceptible genotypes on account of H. armigera herbivory. Expression of resistance to H. armigera was further revealed when significant negative association between NR, NiR, nitrite and percent pod damage was observed. This is the first report that suggests nitrite to be a potent inhibitor of H. armigera α-amylase and also the involvement of nitrite and NR in providing resistance against H. armigera herbivory.

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

    PubMed

    Chen, Chengyu; Liu, Ying; Shi, Xueyan; Desneux, Nicolas; Han, Peng; Gao, Xiwu

    2017-01-01

    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.

  17. A RAPD-PCR-based genetic diversity analysis of Helicoverpa armigera and H. zea populations in Brazil.

    PubMed

    Lopes, H M; Bastos, C S; Boiteux, L S; Foresti, J; Suinaga, F A

    2017-09-21

    Helicoverpa armigera is the most significant pest of agriculture in Asia, Europe, Africa, and Australasia, causing damage to crops greater than US$2 billion annually and until 2013 it was not detected in Brazil. Helicoverpa zea is restricted to the American continent and is important to corn and a secondary pest of cotton and tomatoes. The wide range of crops exploited by H. armigera (mainly cotton, soybeans, chickpea, and corn), the possible mating between these species can promote population shifts, that could be assessed by RAPD-PCR technique. Therefore, the aim of this study was to determine the genetic diversity of H. armigera and H. zea populations by RAPD-PCR analysis. The most important result was the clustering of one H. armigera population in a group predominantly formed by H. zea. It could indicate a possible occurrence of an interspecific cross between these species. This is a concern to Brazilian agriculture due to the possibility of selection of hybrids well adapted to the American environment, which would be inherited from H. zea. The other noxious fact is the possible development of new biotypes resistant to insectides or Bt toxins expressed in transgenic crops, came from H. armigera gene pool.

  18. Way toward "dietary pesticides": molecular investigation of insecticidal action of caffeic acid against Helicoverpa armigera.

    PubMed

    Joshi, R S; Wagh, T P; Sharma, N; Mulani, F A; Sonavane, U; Thulasiram, H V; Joshi, R; Gupta, V S; Giri, A P

    2014-11-12

    Bioprospecting of natural molecules is essential to overcome serious environmental issues and pesticide resistance in insects. Here we are reporting insights into insecticidal activity of a plant natural phenol. In silico and in vitro screening of multiple molecules supported by in vivo validations suggested that caffeic acid (CA) is a potent inhibitor of Helicoverpa armigera gut proteases. Protease activity and gene expression were altered in CA-fed larvae. The structure-activity relationship of CA highlighted that all the functional groups are crucial for inhibition of protease activity. Biophysical studies and molecular dynamic simulations revealed that sequential binding of multiple CA molecules induces conformational changes in the protease(s) and thus lead to a significant decline in their activity. CA treatment significantly inhibits the insect's detoxification enzymes, thus intensifying the insecticidal effect. Our findings suggest that CA can be implicated as a potent insecticidal molecule and explored for the development of effective dietary pesticides.

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

    PubMed Central

    Vergara, Fredd; Shino, Amiu; Kikuchi, Jun

    2016-01-01

    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-13C 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. PMID:27598144

  20. Alterations in the Helicoverpa armigera midgut digestive physiology after ingestion of pigeon pea inducible leucine aminopeptidase.

    PubMed

    Lomate, Purushottam R; Jadhav, Bhakti R; Giri, Ashok P; Hivrale, Vandana K

    2013-01-01

    Jasmonate inducible plant leucine aminopeptidase (LAP) is proposed to serve as direct defense in the insect midgut. However, exact functions of inducible plant LAPs in the insect midgut remain to be estimated. In the present investigation, we report the direct defensive role of pigeon pea inducible LAP in the midgut of Helicoverpa armigera (Lepidoptera: Noctuidae) and responses of midgut soluble aminopeptidases and serine proteinases upon LAP ingestion. Larval growth and survival was significantly reduced on the diets supplemented with pigeon pea LAP. Aminopeptidase activities in larvae remain unaltered in presence or absence of inducible LAP in the diet. On the contrary, serine proteinase activities were significantly decreased in the larvae reared on pigeon pea LAP containing diet as compared to larvae fed on diet without LAP. Our data suggest that pigeon pea inducible LAP is responsible for the degradation of midgut serine proteinases upon ingestion. Reduction in the aminopeptidase activity with LpNA in the H. armigera larvae was compensated with an induction of aminopeptidase activity with ApNA. Our findings could be helpful to further dissect the roles of plant inducible LAPs in the direct plant defense against herbivory.

  1. Alterations in the Helicoverpa armigera Midgut Digestive Physiology after Ingestion of Pigeon Pea Inducible Leucine Aminopeptidase

    PubMed Central

    Lomate, Purushottam R.; Jadhav, Bhakti R.; Giri, Ashok P.; Hivrale, Vandana K.

    2013-01-01

    Jasmonate inducible plant leucine aminopeptidase (LAP) is proposed to serve as direct defense in the insect midgut. However, exact functions of inducible plant LAPs in the insect midgut remain to be estimated. In the present investigation, we report the direct defensive role of pigeon pea inducible LAP in the midgut of Helicoverpa armigera (Lepidoptera: Noctuidae) and responses of midgut soluble aminopeptidases and serine proteinases upon LAP ingestion. Larval growth and survival was significantly reduced on the diets supplemented with pigeon pea LAP. Aminopeptidase activities in larvae remain unaltered in presence or absence of inducible LAP in the diet. On the contrary, serine proteinase activities were significantly decreased in the larvae reared on pigeon pea LAP containing diet as compared to larvae fed on diet without LAP. Our data suggest that pigeon pea inducible LAP is responsible for the degradation of midgut serine proteinases upon ingestion. Reduction in the aminopeptidase activity with LpNA in the H. armigera larvae was compensated with an induction of aminopeptidase activity with ApNA. Our findings could be helpful to further dissect the roles of plant inducible LAPs in the direct plant defense against herbivory. PMID:24098675

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

    PubMed

    Vergara, Fredd; Shino, Amiu; Kikuchi, Jun

    2016-09-02

    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.

  3. Organophosphate and Pyrethroid Hydrolase Activities of Mutant Esterases from the Cotton Bollworm Helicoverpa armigera

    PubMed Central

    Li, Yongqiang; Farnsworth, Claire A.; Coppin, Chris W.; Teese, Mark G.; Liu, Jian-Wei; Scott, Colin; Zhang, Xing; Russell, Robyn J.; Oakeshott, John G.

    2013-01-01

    Two mutations have been found in five closely related insect esterases (from four higher Diptera and a hymenopteran) which each confer organophosphate (OP) hydrolase activity on the enzyme and OP resistance on the insect. One mutation converts a Glycine to an Aspartate, and the other converts a Tryptophan to a Leucine in the enzymes’ active site. One of the dipteran enzymes with the Leucine mutation also shows enhanced activity against pyrethroids. Introduction of the two mutations in vitro into eight esterases from six other widely separated insect groups has also been reported to increase substantially the OP hydrolase activity of most of them. These data suggest that the two mutations could contribute to OP, and possibly pyrethroid, resistance in a variety of insects. We therefore introduced them in vitro into eight Helicoverpa armigera esterases from a clade that has already been implicated in OP and pyrethroid resistance. We found that they do not generally enhance either OP or pyrethroid hydrolysis in these esterases but the Aspartate mutation did increase OP hydrolysis in one enzyme by about 14 fold and the Leucine mutation caused a 4–6 fold increase in activity (more in one case) of another three against some of the most insecticidal isomers of fenvalerate and cypermethrin. The Aspartate enzyme and one of the Leucine enzymes occur in regions of the H. armigera esterase isozyme profile that have been previously implicated in OP and pyrethroid resistance, respectively. PMID:24204917

  4. A sugar gustatory receptor identified from the foregut of cotton bollworm Helicoverpa armigera.

    PubMed

    Xu, Wei; Zhang, Hui-Jie; Anderson, Alisha

    2012-12-01

    Helicoverpa armigera (Hübner) is one of the most polyphagous and cosmopolitan pest species, the larvae of which feed on numerous important crops. The gustatory system is critical in guiding insect feeding behavior. Here, we identified a gustatory receptor from H. armigera, HaGR9, which shows high levels of identity to DmGR43a from Drosophila melanogaster and BmGR9 from Bombyx mori. Reverse transcriptase PCR (RT-PCR) revealed HaGR9 is highly expressed in larval foregut, with little or no expression in other chemosensory tissues. Membrane topology studies indicated that, like two previously studied B. mori GRs, BmGR8 and BmGR53, HaGR9 has an inverted topology relative to G protein-coupled receptors (GPCRs), an intracellular N-terminus and an extracellular C-terminus. Calcium imaging studies confirmed HaGR9 is a sugar receptor showing dose-dependent responses to D-galactose, D-maltose, and D-fructose. This highly-expressed foregut-specific gustatory receptor may contribute to the regulation of larval feeding behavior.

  5. Proteomic and metabolomic profiles of larval hemolymph associated with diapause in the cotton bollworm, Helicoverpa armigera.

    PubMed

    Zhang, Qi; Lu, Yu-Xuan; Xu, Wei-Hua

    2013-11-01

    Diapause is programmed developmental arrest coupled with the depression of metabolic activity and the enhancement of stress resistance. Pupal diapause is induced by environmental signals and is prepared during the prediapause phase. In the cotton bollworm, Helicoverpa armigera, the prediapause phase, which contains two sub-phases, diapause induction and preparation, occurs in the larval stage. Here, we performed parallel proteomic and metabolomic analyses on H. armigera larval hemolymph during the prediapause phase. By two-dimensional electrophoresis, 37 proteins were shown to be differentially expressed in diapause-destined larvae. Of these proteins, 28 were successfully identified by MALDI-TOF/TOF mass spectrometry. Moreover, a total of 22 altered metabolites were found in diapause-destined larval hemolymph by GC-MS analysis, and the levels of 17 metabolites were elevated and 5 were decreased. The proteins and metabolites with significantly altered levels play different roles in diapause-destined larvae, including diapause induction, metabolic storage, immune response, stress tolerance, and others. Because hemolymph circulates through the whole body of an insect, these differences found in diapause-destined larvae most likely correspond to upstream endocrine signals and would further influence other organ/tissue activities to determine the insect's fact: diapause or development.

  6. Organophosphate and pyrethroid hydrolase activities of mutant Esterases from the cotton bollworm Helicoverpa armigera.

    PubMed

    Li, Yongqiang; Farnsworth, Claire A; Coppin, Chris W; Teese, Mark G; Liu, Jian-Wei; Scott, Colin; Zhang, Xing; Russell, Robyn J; Oakeshott, John G

    2013-01-01

    Two mutations have been found in five closely related insect esterases (from four higher Diptera and a hymenopteran) which each confer organophosphate (OP) hydrolase activity on the enzyme and OP resistance on the insect. One mutation converts a Glycine to an Aspartate, and the other converts a Tryptophan to a Leucine in the enzymes' active site. One of the dipteran enzymes with the Leucine mutation also shows enhanced activity against pyrethroids. Introduction of the two mutations in vitro into eight esterases from six other widely separated insect groups has also been reported to increase substantially the OP hydrolase activity of most of them. These data suggest that the two mutations could contribute to OP, and possibly pyrethroid, resistance in a variety of insects. We therefore introduced them in vitro into eight Helicoverpa armigera esterases from a clade that has already been implicated in OP and pyrethroid resistance. We found that they do not generally enhance either OP or pyrethroid hydrolysis in these esterases but the Aspartate mutation did increase OP hydrolysis in one enzyme by about 14 fold and the Leucine mutation caused a 4-6 fold increase in activity (more in one case) of another three against some of the most insecticidal isomers of fenvalerate and cypermethrin. The Aspartate enzyme and one of the Leucine enzymes occur in regions of the H. armigera esterase isozyme profile that have been previously implicated in OP and pyrethroid resistance, respectively.

  7. Nutritional performance of the tomato fruit borer, Helicoverpa armigera, on different tomato cultivars.

    PubMed

    Kouhi, Davoud; Naseri, Bahram; Golizadeh, Ali

    2014-01-01

    The development and cultivation of tomato cultivars that are resistant to the tomato fruit borer, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), are very limited in Iran and other parts of the world because of the lack of information about resistant tomato cultivars to minimize the use of insecticides. Therefore, the present study was carried out to identify alternative methods to chemical control. Nutritional performance of the larval stages (fourth, fifth, and sixth instars) of H. armigera on fruit of eight tomato cultivars, including SUN 6108 f1, Rio grande UG, Korral, Super strain B, CH falat, Hed rio grande, Cal.JN3, and Super crystal, was studied under laboratory conditions. Fourth instars reared on CH falat and SUN 6108 f1 respectively showed the highest and lowest values of approximate digestibility. The highest values of efficiency of conversion of ingested food and efficiency of conversion of digested food of fifth instars were on Super strain B. The relative consumption rate and relative growth rate values of the sixth instars were the highest on Korral. The highest and lowest values of consumption index of sixth instars were on Super strain B and Hed rio grande, respectively. The efficiency of conversion of ingested food and efficiency of conversion of digested food values of whole larval instars were the highest on Hed rio grande and lowest on Rio grande UG. The results of nutritional indices indicated that Rio grande UG is an unsuitable host for H. armigera. This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.

  8. In-Plant Protection against Helicoverpa armigera by Production of Long hpRNA in Chloroplasts

    PubMed Central

    Bally, Julia; McIntyre, Glen J.; Doran, Rachel L.; Lee, Karen; Perez, Alicia; Jung, Hyungtaek; Naim, Fatima; Larrinua, Ignacio M.; Narva, Kenneth E.; Waterhouse, Peter M.

    2016-01-01

    Expressing double-stranded RNA (dsRNA) in transgenic plants to silence essential genes within herbivorous pests is referred to as trans-kingdom RNA interference (TK-RNAi) and has emerged as a promising strategy for crop protection. However, the dicing of dsRNA into siRNAs by the plant’s intrinsic RNAi machinery may reduce this pesticidal activity. Therefore, genetic constructs, encoding ∼200 nt duplex-stemmed-hairpin (hp) RNAs, targeting the acetylcholinesterase gene of the cotton bollworm, Helicoverpa armigera, were integrated into either the nuclear or the chloroplast genome of Nicotiana benthamiana. Undiced, full-length hpRNAs accumulated in transplastomic lines of N. benthamiana and conferred strong protection against H. armigera herbivory while the hpRNAs of nuclear-transformed plants were processed into siRNAs and gave more modest anti-feeding activity. This suggests that there is little or no RNAi machinery or activity in the chloroplast, that hpRNAs produced within this organelle do not enter the cytoplasm, and that oral delivery of chloroplast-packaged intact hpRNA is a more effective means of delivering TK-RNAi than using nuclear encoded hpRNAs. This contrasts with a recently reported correlation between siRNA expression and effectiveness of TK-RNAi targeting the chitinase gene of H. armigera, but is consistent with reports of efficient TK-RNAi by dsRNA generated in chloroplasts by converging promoters flanking a pest gene sequence and from very small (21 nt-stem) hpRNAs resembling artificial miRNAs. Here we demonstrate that hpRNAs, constructed along the conventional design principles of plant RNAi constructs but integrated into the chloroplast genome, are stable and effective over multiple generations, and hold the promise of providing durable pest resistance in crops. PMID:27746796

  9. RNAi of selected candidate genes interrupts growth and development of Helicoverpa armigera.

    PubMed

    Chikate, Yojana R; Dawkar, Vishal V; Barbole, Ranjit S; Tilak, Priyadarshini V; Gupta, Vidya S; Giri, Ashok P

    2016-10-01

    Helicoverpa armigera is one of the major crop pests and is less amenable to current pest control approaches. RNA interference (RNAi) is emerging as a potent arsenal for the insect pest control over current methods. Here, we examined the effect on growth and development in H. armigera by targeting various enzymes/proteins such as proteases like trypsins (HaTry2, 3, 4 and 6), chymotrypsin (HaChy4) and cysteine protease like cathepsin (HaCATHL); glutathione S-transferases (HaGST1a, 6 and 8); esterases (HaAce4, HaJHE); catalase (HaCAT); super-oxide-dismutase (HaCu/ZnSOD); fatty acid binding protein (HaFabp) and chitin deacetylase (HaCda5b) through dsRNA approach. Significant downregulation of cognate mRNA expression and reduced activity of trypsin and GST-like enzyme were evident upon feeding candidate dsRNAs to the larvae. Among these, the highest mortality was observed in HaAce4 dsRNA fed larvae followed by HaJHE; HaCAT; HaCuZnSOD; HaFabp and HaTry3 whereas remaining ones showed relatively lower mortality. Furthermore, the dsRNA fed larvae showed significant reduction in the larval mass and abnormalities at the different stages of H. armigera development compared to their control diets. For example, malformed larvae, pupae and moth at a dose of 60μg/day were evident in high number of individual insects fed on dsRNA containing diets. Moreover, the growth and development of insects and moths were retarded in dsRNA fed larvae. These findings might provide potential new candidates for designing effective dsRNA as pesticide in crop protection. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. In-Plant Protection against Helicoverpa armigera by Production of Long hpRNA in Chloroplasts.

    PubMed

    Bally, Julia; McIntyre, Glen J; Doran, Rachel L; Lee, Karen; Perez, Alicia; Jung, Hyungtaek; Naim, Fatima; Larrinua, Ignacio M; Narva, Kenneth E; Waterhouse, Peter M

    2016-01-01

    Expressing double-stranded RNA (dsRNA) in transgenic plants to silence essential genes within herbivorous pests is referred to as trans-kingdom RNA interference (TK-RNAi) and has emerged as a promising strategy for crop protection. However, the dicing of dsRNA into siRNAs by the plant's intrinsic RNAi machinery may reduce this pesticidal activity. Therefore, genetic constructs, encoding ∼200 nt duplex-stemmed-hairpin (hp) RNAs, targeting the acetylcholinesterase gene of the cotton bollworm, Helicoverpa armigera, were integrated into either the nuclear or the chloroplast genome of Nicotiana benthamiana. Undiced, full-length hpRNAs accumulated in transplastomic lines of N. benthamiana and conferred strong protection against H. armigera herbivory while the hpRNAs of nuclear-transformed plants were processed into siRNAs and gave more modest anti-feeding activity. This suggests that there is little or no RNAi machinery or activity in the chloroplast, that hpRNAs produced within this organelle do not enter the cytoplasm, and that oral delivery of chloroplast-packaged intact hpRNA is a more effective means of delivering TK-RNAi than using nuclear encoded hpRNAs. This contrasts with a recently reported correlation between siRNA expression and effectiveness of TK-RNAi targeting the chitinase gene of H. armigera, but is consistent with reports of efficient TK-RNAi by dsRNA generated in chloroplasts by converging promoters flanking a pest gene sequence and from very small (21 nt-stem) hpRNAs resembling artificial miRNAs. Here we demonstrate that hpRNAs, constructed along the conventional design principles of plant RNAi constructs but integrated into the chloroplast genome, are stable and effective over multiple generations, and hold the promise of providing durable pest resistance in crops.

  11. Draft Genome Sequence of Bacillus thuringiensis Serovar Tolworthi Strain Na205-3, an Isolate Toxic for Helicoverpa armigera

    PubMed Central

    Palma, Leopoldo; Muñoz, Delia; Murillo, Jesús

    2014-01-01

    We report here the complete annotated 6,510,053-bp draft genome sequence of Bacillus thuringiensis serovar tolworthi strain Na205-3, which is toxic for Helicoverpa armigera. This strain potentially contains nine insecticidal toxin genes homologous to cry1Aa12, cry1Ab1, cry1Ab8, cry1Ba1, cry1Af1, cry1Ia10, vip1Bb1, vip2Ba2, and vip3Aa6. PMID:24625875

  12. Specific binding of activated Vip3Aa10 to Helicoverpa armigera brush border membrane vesicles results in pore formation.

    PubMed

    Liu, Jing-Guo; Yang, Ai-Zhen; Shen, Xiao-Hong; Hua, Bao-Guang; Shi, Guang-Lu

    2011-10-01

    Helicoverpa armigera is one of the most harmful pests in China. Although it had been successfully controlled by Cry1A toxins, some H. armigera populations are building up resistance to Cry1A toxins in the laboratory. Vip3A, secreted by Bacillus thuringiensis, is another potential toxin against H. armigera. Previous reports showed that activated Vip3A performs its function by inserting into the midgut brush border membrane vesicles (BBMV) of susceptible insects. To further investigate the binding of Vip3A to BBMV of H. armigera, the full-length Vip3Aa10 toxin expressed in Escherichia coli was digested by trypsin or midgut juice extract, respectively. Among the fragments of digested Vip3Aa10, only a 62kDa fragment (Vip3Aa10-T) exhibited binding to BBMV of H. armigera and has insecticidal activity. Moreover, this interaction was specific and was not affected by the presence of Cry1Ab toxin. Binding of Vip3Aa10-T to BBMV resulted in the formation of an ion channel. Unlike Cry1A toxins, Vip3Aa10-T was just slightly associated with lipid rafts of BBMV. These data suggest that although activated Vip3Aa10 specifically interacts with BBMV of H. armigera and forms an ion channel, the mode of action of it may be different from that of Cry1A toxins.

  13. Feeding deterrent and growth inhibitory activities of PONNEEM, a newly developed phytopesticidal formulation against Helicoverpa armigera (Hubner)

    PubMed Central

    Packiam, Soosaimanickam Maria; Baskar, Kathirvelu; Ignacimuthu, Savarimuthu

    2014-01-01

    Objective To assess the feeding deterrent, growth inhibitory and egg hatchability effects of PONNEEM on Helicoverpa armigera (H. armigera). Methods Five oil formulations were prepared at different ratios to assess the feeding deterrent, growth inhibitory and egg hatchability effects on H. armigera. Results Invariably all the newly formulated phytopesticidal oil formulations showed the feeding deterrent and growth inhibitory activities against H. armigera. The maximum feeding deterrent activity of 88.44% was observed at 15 µL/L concentration of PONNEEM followed by formulation A (74.54%). PONNEEM was found to be effective in growth inhibitory activities and egg hatchability at 10 µL/L concentration. It exhibited statistically significant feeding deterrent activity and growth inhibitory activity compared with all the other treatments. Conclusions PONNEEM was found to be effective phytopesticidal formulation to control the larval stage of H. armigera. This is the first report for the feeding deterrent activity of PONNEEM against H. armigera. This newly formulated phytopesticide was patented in India. PMID:25183105

  14. Variation in susceptibility of Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) (Lepidoptera: Noctuidae) in Australia to two Bacillus thuringiensis toxins.

    PubMed

    Bird, Lisa J; Akhurst, Raymond J

    2007-02-01

    Intra-specific variation in susceptibility of Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) in Australia to the Cry1Ac and Cry2Ab delta-endotoxins from Bacillus thuringiensis (Berliner) (Bt) was determined to establish a baseline for monitoring changes that might occur with the use of Bt cotton. Strains of H. armigera and H. punctigera were established from populations collected primarily from commercial farms throughout the Australian cotton belts. Strains were evaluated for susceptibility using two bioassay methods (surface treatment and diet incorporation) by measuring the dose response for mortality (LC50) and growth inhibition (IC50). The variation in LC50 among H. armigera (n=17 strains) and H. punctigera (n=12 strains) in response to Cry1Ac was 4.6- and 3.2-fold, respectively. The variation in LC50 among H. armigera (n=19 strains) and H. punctigera (n=12 strains) to Cry2Ab was 6.6- and 3.5-fold, respectively. The range of Cry1Ac induced growth inhibition from the 3rd to 4th instar in H. armigera (n=15 strains) was 3.6-fold and in H. punctigera (n=13 strains) was 2.6-fold, while the range of Cry2Ab induced growth inhibition from neonate to 3rd instar in H. armigera (n=13 strains) was 4.3-fold and in H. punctigera (n=12 strains) was 6.1-fold. Variation in susceptibility was also evaluated for two age classes (neonates and 3rd instars) in laboratory strains of H. armigera and H. punctigera. Neonates of H. punctigera had the same or higher sensitivity to Bt than 3rd instars. Neonates of H. armigera were more sensitive to Cry2Ab than 3rd instars, while being less sensitive to Cry1Ac than 3rd instars. Differences in the two methods of bioassay used affected relative sensitivity of species to Bt toxins, highlighting the need to standardize bioassay protocols.

  15. Interaction of plant cell signaling molecules, salicylic acid and jasmonic acid, with the mitochondria of Helicoverpa armigera.

    PubMed

    Akbar, S M D; Sharma, H C; Jayalakshmi, S K; Sreeramulu, K

    2012-02-01

    The cotton bollworm, Helicoverpa armigera is a polyphagous pest in Asia, Africa, and the Mediterranean Europe. Salicylic acid (SA) and jasmonic acid (JA) are the cell signaling molecules produced in response to insect attack in plants. The effect of these signaling molecules was investigated on the oxidative phosphorylation and oxidative stress of H. armigera. SA significantly inhibited the state III and state IV respiration, respiratory control index (RCI), respiratory complexes I and II, induced mitochondrial swelling, and cytochrome c release in vitro. Under in vivo conditions, SA induced state IV respiration as well as oxidative stress in time- and dose-dependent manner, and also inhibited the larval growth. In contrast, JA did not affect the mitochondrial respiration and oxidative stress. SA affected the growth and development of H. armigera, in addition to its function as signaling molecules involved in both local defense reactions at feeding sites and the induction of systemic acquired resistance in plants.

  16. Cross-species amplification and polymorphism of microsatellite loci in Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) in Brazilian cropping systems.

    PubMed

    Leite, N A; Corrêa, A S; Alves-Pereira, A; Campos, J B; Zucchi, M I; Omoto, C

    2016-04-04

    The Old World bollworm Helicoverpa armigera (Hübner) was recently discovered in Brazil. This species is closely related to the New World bollworm H. zea (Boddie), and mating between these species has already been reported under laboratory conditions. Here, we tested the cross-species amplification of 20 microsatellite (SSR) loci in field populations of H. armigera and H. zea collected from Brazilian cropping systems. Seven SSR loci were successfully amplified and polymorphic in both species except for the locus HaC14, which was monomorphic for H. zea. All SSR loci were in linkage equilibrium, and deviations from Hardy- Weinberg equilibrium were only observed for the locus HarSSR1 in the HaRS-2 population, where null alleles were present. A moderate level of polymorphism was detected in H. armigera and H. zea populations with a mean allele number of 4.14, and 2.24, respectively. Interestingly, most of the populations of the recent invader H. armigera showed higher genetic diversity and inbreeding coefficients than H. zea populations. The genetic identity of each species was recovered using a STRUCTURE analysis, where the populations formed two clusters (K = 2) according to their species. STRUCTURE also suggested the occurrence of potential hybrid offspring between H. armigera and H. zea individuals in natural conditions. These SSR loci will be valuable in characterizing population differentiation, invasion routes, adaptation, reproductive behavior, and intra- and interspecific gene flow in H. armigera and H. zea populations in Brazil, the USA, and other areas where these two pests occur.

  17. Targeting chitinase gene of Helicoverpa armigera by host-induced RNA interference confers insect resistance in tobacco and tomato.

    PubMed

    Mamta; Reddy, K R K; Rajam, M V

    2016-02-01

    Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a devastating agricultural insect pest with broad spectrum of host range, causing million dollars crop loss annually. Limitations in the present conventional and transgenic approaches have made it crucial to develop sustainable and environmental friendly methods for crop improvement. In the present study, host-induced RNA interference (HI-RNAi) approach was used to develop H. armigera resistant tobacco and tomato plants. Chitinase (HaCHI) gene, critically required for insect molting and metamorphosis was selected as a potential target. Hair-pin RNAi construct was prepared from the conserved off-target free partial HaCHI gene sequence and was used to generate several HaCHI-RNAi tobacco and tomato plants. Northern hybridization confirmed the production of HaCHI gene-specific siRNAs in HaCHI-RNAi tobacco and tomato lines. Continuous feeding on leaves of RNAi lines drastically reduced the target gene transcripts and consequently, affected the overall growth and survival of H. armigera. Various developmental deformities were also manifested in H. armigera larvae after feeding on the leaves of RNAi lines. These results demonstrated the role of chitinase in insect development and potential of HI-RNAi for effective management of H. armigera.

  18. The expression of proteins involved in digestion and detoxification are regulated in Helicoverpa armigera to cope up with chlorpyrifos insecticide.

    PubMed

    Dawkar, Vishal V; Chikate, Yojana R; More, Tushar H; Gupta, Vidya S; Giri, Ashok P

    2016-02-01

    Helicoverpa armigera is a key pest in many vital crops, which is mainly controlled by chemical strategies. To manage this pest is becoming challenging due to its ability and evolution of resistance against insecticides. Further, its subsequent spread on nonhost plant is remarkable in recent times. Hence, decoding resistance mechanism against phytochemicals and synthetic insecticides is a major challenge. The present work describes that the digestion, defense and immunity related enzymes are associated with chlorpyrifos resistance in H. armigera. Proteomic analysis of H. armigera gut tissue upon feeding on chlorpyrifos containing diet (CH) and artificial diet (AD) using nano-liquid chromatography-mass spectrometry identified upregulated 23-proteins in CH fed larvae. Database searches combined with gene ontology analysis revealed that the identified gut proteins engrossed in digestion, proteins crucial for immunity, adaptive responses to stress, and detoxification. Biochemical and quantitative real-time polymerase chain reaction analysis of candidate proteins indicated that insects were struggling to get nutrients and energy in presence of CH, while at the same time endeavoring to metabolize chlorpyrifos. Moreover, we proposed a potential processing pathway of chlorpyrifos in H. armigera gut by examining the metabolites using gas chromatography-mass spectrometry. H. armigera exhibit a range of intriguing behavioral, morphological adaptations and resistance to insecticides by regulating expression of proteins involved in digestion and detoxification mechanisms to cope up with chlorpyrifos. In these contexts, as gut is a rich repository of biological information; profound analysis of gut tissues can give clues of detoxification and resistance mechanism in insects.

  19. Induced resistance to Helicoverpa armigera through exogenous application of jasmonic acid and salicylic acid in groundnut, Arachis hypogaea.

    PubMed

    War, Abdul Rashid; Paulraj, Michael Gabriel; Ignacimuthu, Savarimuthu; Sharma, Hari Chand

    2015-01-01

    Induced resistance to Helicoverpa armigera through exogenous application of jasmonic acid (JA) and salicylic acid (SA) was studied in groundnut genotypes (ICGV 86699, ICGV 86031, ICG 2271 and ICG 1697) with different levels of resistance to insects and the susceptible check JL 24 under greenhouse conditions. Activities of oxidative enzymes and the amounts of secondary metabolites and proteins were quantified at 6 days after JA and SA application/insect infestation. Data were also recorded on plant damage and H. armigera larval weights and survival. Higher levels of enzymatic activities and amounts of secondary metabolites were observed in the insect-resistant genotypes pretreated with JA and then infested with H. armigera than in JL 24. The insect-resistant genotypes suffered lower insect damage and resulted in poor survival and lower weights of H. armigera larvae than JL 24. In some cases, JA and SA showed similar effects. JA and SA induced the activity of antioxidative enzymes in groundnut plants against H. armigera, and reduced its growth and development. However, induced response to application of JA was greater than to SA, and resulted in reduced plant damage, and larval weights and survival, suggesting that induced resistance can be used as a component of pest management in groundnut. © 2014 Society of Chemical Industry.

  20. Molecular Cloning, Expression, and Identification of Bre Genes Involved in Glycosphingolipids Synthesis in Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Zhang, Dandan; Xiao, Yutao; Hussain Dhiloo, Khalid; Soberon, Mario; Bravo, Alejandra; Wu, Kongming

    2016-05-17

    Glycosphingolipids (GSLs) play important roles in the cellular biology of vertebrate and invertebrate organisms, such as cell differentiation, tumor metastasis, and cell coordination. GSLs also serve as receptors for different bacterial toxins. For example, in the nematode Caenorhabditis elegans, GSLs function as receptors of the insecticidal Cry toxins produced by Bacillus thuringiensis (Bt), and mutations in bre genes involved in GSLs synthesis resulted in resistance to Cry5 toxin in this organism. However, the information of GSLs function in insects is still limited. In this study, three genes for glycosyltransferases, bre2, bre3, and bre4, from Helicoverpa armigera were identified and cloned. The previously reported bre5 gene from H. armigera was also analyzed. Protein sequence alignments revealed that proteins codified by H. armigera Bre shared high identity with homologous proteins from other organisms. Expression profile analysis revealed that the expressions of bre genes varied in the different tissues and also in the different developmental stages of H. armigera. Finally, the heterologous expression of bre genes in Trichoplusia ni Hi5 cell line showed that the corresponding translated proteins were localized in the cytoplasm of Hi5 cells. These results provide the bases for further functional studies of bre genes and analyzing potential roles of GSLs in mode of action of Cry1A toxin in H. armigera.

  1. Interaction of Bacillus thuringiensis Toxins with Larval Midgut Binding Sites of Helicoverpa armigera (Lepidoptera: Noctuidae)

    PubMed Central

    Estela, Anna; Escriche, Baltasar; Ferré, Juan

    2004-01-01

    In 1996, Bt-cotton (cotton expressing a Bacillus thuringiensis toxin gene) expressing the Cry1Ac protein was commercially introduced to control cotton pests. A threat to this first generation of transgenic cotton is the evolution of resistance by the insects. Second-generation Bt-cotton has been developed with either new B. thuringiensis genes or with a combination of cry genes. However, one requirement for the “stacked” gene strategy to work is that the stacked toxins bind to different binding sites. In the present study, the binding of 125I-labeled Cry1Ab protein (125I-Cry1Ab) and 125I-Cry1Ac to brush border membrane vesicles (BBMV) of Helicoverpa armigera was analyzed in competition experiments with 11 nonlabeled Cry proteins. The results indicate that Cry1Aa, Cry1Ab, and Cry1Ac competed for common binding sites. No other Cry proteins tested competed for either 125I-Cry1Ab or 125I-Cry1Ac binding, except Cry1Ja, which competed only at the highest concentrations used. Furthermore, BBMV from four H. armigera populations were also tested with 125I-Cry1Ac and Cry1Ab to check the influence of the insect population on the binding results. Finally, the inhibitory effect of selected sugars and lectins was also determined. 125I-Cry1Ac binding was strongly inhibited by N-acetylgalactosamine, sialic acid, and concanavalin A and moderately inhibited by soybean agglutinin. In contrast, 125I-Cry1Ab binding was only significantly inhibited by concanavalin A. These results show that Cry1Ac and Cry1Ab use different epitopes for binding to BBMV. PMID:15006756

  2. Effect of different host plants on nutritional indices of the pod borer, Helicoverpa armigera.

    PubMed

    Hemati, S A; Naseri, B; Ganbalani, G Nouri; Dastjerdi, H Rafiee; Golizadeh, A

    2012-01-01

    Nutritional indices of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) on different host plants including chickpea (cultivars Arman, Hashem, Azad, and Binivich), common bean (cultivar Khomein), white kidney bean (cultivar Dehghan), red kidney bean (cultivar Goli), cowpea (cultivar Mashhad), tomato (cultivar Meshkin) and potato (cultivars Agria and Satina) were studied under laboratory conditions (25 ± 1 °C, 65 ± 5% RH, 16:8 L:D). Third instar larvae reared on potato Agria showed the highest efficiency of conversion of digested food (ECD) and efficiency of conversion of ingested food (ECI) (50.800 ± 0.104% and 13.630 ± 0.016%, respectively). Approximate digestibility (AD) values of the fourth instar larvae were highest (92.651 ± 0.004%) and lowest (57.140 - 0.049%) on chickpea Azad and potato Agria, respectively. The fifth instar larvae fed on tomato Meshkin and white kidney bean Dehghan had the highest consumption index (CI) (3.717 ± 0.091) and relative consumption rate (RCR) (1.620 ± 0.074), respectively. Whole larval instars showed the highest ECI and ECD values on potatoes Satina (14.640 ± 0.014%) and Agria (21.380 ± 0.015%), respectively, and the lowest of both values on tomato Meshkin (ECI: 5.748 ± 0.002% and ECD: 7.341 ± 0.002%). The results of nutritional indices and the cluster analysis indicated that tomato Meshkin was an unsuitable host for feeding of H. armigera.

  3. Effect of electron beam irradiation on developmental stages of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

    NASA Astrophysics Data System (ADS)

    Kim, Junheon; Chung, Soon-Oh; Jang, Sin Ae; Jang, Miyeon; Park, Chung Gyoo

    2015-07-01

    Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), is an economically important and polyphagous pest, which harms various kinds of food crops and important agricultural plants, such as cotton and paprika. Effects of electron beam irradiation at six dose levels between 50 and 350 Gy on the egg (24-48 h old), the larval (4-5th instar), and the pupal (7-d old for female, 5-d old for male) development, and on the adult (1-d old) reproduction were tested to identify a potential quarantine treatment dose. Increased doses of irradiation on eggs decreased egg hatchability, pupation and adult emergence and increased larval period. ED99 values for inhibition of hatching, pupation and emergence were 460.6, 236.9 and 197.8 Gy, respectively. When larvae were irradiated with more than 280 Gy, no larvae could develop into pupae. ED99 values for inhibition of pupation and adult emergence were 265.6 and 189.6 Gy, respectively. Even though the irradiation on pupa did not completely inhibit adult emergence, most of the pupae emerged to deformed adults. When adults were irradiated, fecundity was not affected. However, F1 egg hatching was completely inhibited at the dose of 350 Gy. ED99 value for inhibition of adult emergence was estimated at 366.5 Gy. Our results suggest that electron beam irradiation could be recommendable as an alternative to MB and as a phytosanitary treatment for quarantine. A treatment dose of less than or equal to 220 Gy is suggested as a potential quarantine treatment to H. armigera egg for prevention of pupation and to larva for prevention of adult emerge.

  4. A droplet digital PCR (ddPCR) assay to detect Helicoverpa armigera (Lepidoptera: Noctuidae) in bulk trap samples

    PubMed Central

    Tembrock, Luke R.; Timm, Alicia E.; Farris, Roxanne E.; Perera, Omaththage P.; Gilligan, Todd M.

    2017-01-01

    Moths in the genus Helicoverpa are some of the most important agricultural pests in the world. Two species, H. armigera (Hübner) and H. zea (Boddie), cause the majority of damage to crops and millions of dollars are spent annually on control of these pests. The recent introduction of H. armigera into the New World has prompted extensive survey efforts for this species in the United States. Surveys are conducted using bucket traps baited with H. armigera pheromone, and, because the same pheromone compounds attract both species, these traps often capture large numbers of the native H. zea. Adult H. armigera and H. zea are very similar and can only be separated morphologically by minor differences in the genitalia. Thus, a time consuming genitalic dissection by a trained specialist is necessary to reliably identify either species, and every specimen must be dissected. Several molecular methods are available for differentiating and identifying H. armigera and H. zea, including two recently developed rapid protocols using real-time PCR. However, none of the published methods are capable of screening specimens in large batches. Here we detail a droplet digital PCR (ddPCR) assay that is capable of detecting a single H. armigera in a background of up to 999 H. zea. The assay has been tested using bulk extractions of 1,000 legs from actual trap samples and is effective even when using poor quality samples. This study provides an efficient, rapid, reproducible, and scalable method for processing H. armigera survey trap samples in the U.S. and demonstrates the potential for applying ddPCR technology to screen and diagnose invasive species. PMID:28562660

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

    PubMed Central

    Khan, Rashid Ahmed; Liu, Ji Yuan; Rashid, Maryam; Wang, Dun; Zhang, Ya Lin

    2013-01-01

    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. PMID:23528854

  6. NMR structure and function of Helicoverpa armigera sterol carrier protein-2, an important insecticidal target from the cotton bollworm.

    PubMed

    Ma, Haihao; Ma, Yuemin; Liu, Xuehui; Dyer, David H; Xu, Pingyong; Liu, Kaiyu; Lan, Que; Hong, Huazhu; Peng, Jianxin; Peng, Rong

    2015-12-10

    The cotton bollworm, Helicoverpa armigera, has developed strong resistance to many insecticides. Sterol Carrier Protein-2 (SCP-2) is an important non-specific lipid transfer protein in insects and appears to be a potential new target. In order to elucidate the structure and function of Helicoverpa armigera SCP-2 (HaSCP-2), NMR spectroscopy, docking simulations, mutagenesis and bioassays were performed. HaSCP-2 composed of five α-helices and four stranded β-sheets. The folds of α-helices and β-sheets interacted together to form a hydrophobic cavity with putative entrance and exit openings, which served as a tunnel for accommodating and transporting of lipids. Several sterols and fatty acids could interact with HaSCP-2 via important hydrophobic sites, which could be potential targets for insecticides. Mutagenesis experiments indicated Y51, F53, F89, F110, I117 and Q131 may be the key functional sites. HaSCP-2 showed high cholesterol binding activity and SCP-2 inhibitors (SCPIs) could inhibit the biological activity of HaSCP-2. SCPI-treated larvae at young stage showed a significant decrease of cholesterol uptake in vivo. Our study describes for the first time a NMR structure of SCP-2 in lepidopteran H. armigera and reveals its important function in cholesterol uptake, which facilitates the screening of effective insecticides targeting the insect cholesterol metabolism.

  7. A TRPA1 channel that senses thermal stimulus and irritating chemicals in Helicoverpa armigera.

    PubMed

    Wei, J J; Fu, T; Yang, T; Liu, Y; Wang, G R

    2015-08-01

    Sensing and responding to changes in the external environment is important for insect survival. Transient receptor potential (TRP) channels are crucial for various sensory modalities including olfaction, vision, hearing, thermosensation and mechanosensation. Here, we identified and characterized a transient receptor potential gene named as HarmTRPA1 in Helicoverpa armigera antennae. HarmTRPA1 was abundantly expressed in the antennae and labial palps. Transcripts of HarmTRPA1 could also be detected in the head and proboscis. Furthermore, functional analyses of HarmTRPA1 were conducted in the Xenopus Oocyte system. The results showed that the HarmTRPA1 channel could be activated by increasing the temperature from 20 to 45 °C. No significant adaptation was observed when the stimulus was repeated. In addition to thermal stimuli, pungent natural compounds including allyl isothiocyanate, cinnamaldehyde and citronellal also activated HarmTRPA1. Taken together, we infer that HarmTRPA1 may function as both a thermal sensor involved in peripheral temperature detection and as a chemical sensor detecting irritating chemicals in vivo. Our data provide valuable insight into the TRPA1 channel in this moth and lay the foundation for developing novel strategies for pest control. © 2015 The Royal Entomological Society.

  8. Mitochondrial DNA and trade data support multiple origins of Helicoverpa armigera (Lepidoptera, Noctuidae) in Brazil

    PubMed Central

    Tay, Wee Tek; Walsh, Thomas K.; Downes, Sharon; Anderson, Craig; Jermiin, Lars S.; Wong, Thomas K. F.; Piper, Melissa C.; Chang, Ester Silva; Macedo, Isabella Barony; Czepak, Cecilia; Behere, Gajanan T.; Silvie, Pierre; Soria, Miguel F.; Frayssinet, Marie; Gordon, Karl H. J.

    2017-01-01

    The Old World bollworm Helicoverpa armigera is now established in Brazil but efforts to identify incursion origin(s) and pathway(s) have met with limited success due to the patchiness of available data. Using international agricultural/horticultural commodity trade data and mitochondrial DNA (mtDNA) cytochrome oxidase I (COI) and cytochrome b (Cyt b) gene markers, we inferred the origins and incursion pathways into Brazil. We detected 20 mtDNA haplotypes from six Brazilian states, eight of which were new to our 97 global COI-Cyt b haplotype database. Direct sequence matches indicated five Brazilian haplotypes had Asian, African, and European origins. We identified 45 parsimoniously informative sites and multiple substitutions per site within the concatenated (945 bp) nucleotide dataset, implying that probabilistic phylogenetic analysis methods are needed. High diversity and signatures of uniquely shared haplotypes with diverse localities combined with the trade data suggested multiple incursions and introduction origins in Brazil. Increasing agricultural/horticultural trade activities between the Old and New Worlds represents a significant biosecurity risk factor. Identifying pest origins will enable resistance profiling that reflects countries of origin to be included when developing a resistance management strategy, while identifying incursion pathways will improve biosecurity protocols and risk analysis at biosecurity hotspots including national ports. PMID:28350004

  9. Characterization of protein phosphatase 5 from three lepidopteran insects: Helicoverpa armigera, Mythimna separata and Plutella xylostella.

    PubMed

    Chen, Xi'en; Lü, Shumin; Zhang, Yalin

    2014-01-01

    Protein phosphatase 5 (PP5), a unique member of serine/threonine phosphatases, regulates a variety of biological processes. We obtained full-length PP5 cDNAs from three lepidopteran insects, Helicoverpa armigera, Mythimna separata and Plutella xylostella, encoding predicted proteins of 490 (55.98 kDa), 490 (55.82 kDa) and 491 (56.07 kDa) amino acids, respectively. These sequences shared a high identity with other insect PP5s and contained the TPR (tetratricopeptide repeat) domains at N-terminal regions and highly conserved C-terminal catalytic domains. Tissue- and stage-specific expression pattern analyses revealed these three PP5 genes were constitutively expressed in all stages and in tested tissues with predominant transcription occurring at the egg and adult stages. Activities of Escherichia coli-produced recombinant PP5 proteins could be enhanced by almost 2-fold by a known PP5 activator: arachidonic acid. Kinetic parameters of three recombinant proteins against substrate pNPP were similar both in the absence or presence of arachidonic acid. Protein phosphatases inhibitors, okadaic acid, cantharidin, and endothall strongly impeded the activities of the three recombinant PP5 proteins, as well as exerted an inhibitory effect on crude protein phosphatases extractions from these three insects. In summary, lepidopteran PP5s share similar characteristics and are all sensitive to the protein phosphatases inhibitors. Our results also imply protein phosphatase inhibitors might be used in the management of lepidopteran pests.

  10. Mitochondrial DNA and trade data support multiple origins of Helicoverpa armigera (Lepidoptera, Noctuidae) in Brazil

    NASA Astrophysics Data System (ADS)

    Tay, Wee Tek; Walsh, Thomas K.; Downes, Sharon; Anderson, Craig; Jermiin, Lars S.; Wong, Thomas K. F.; Piper, Melissa C.; Chang, Ester Silva; Macedo, Isabella Barony; Czepak, Cecilia; Behere, Gajanan T.; Silvie, Pierre; Soria, Miguel F.; Frayssinet, Marie; Gordon, Karl H. J.

    2017-03-01

    The Old World bollworm Helicoverpa armigera is now established in Brazil but efforts to identify incursion origin(s) and pathway(s) have met with limited success due to the patchiness of available data. Using international agricultural/horticultural commodity trade data and mitochondrial DNA (mtDNA) cytochrome oxidase I (COI) and cytochrome b (Cyt b) gene markers, we inferred the origins and incursion pathways into Brazil. We detected 20 mtDNA haplotypes from six Brazilian states, eight of which were new to our 97 global COI-Cyt b haplotype database. Direct sequence matches indicated five Brazilian haplotypes had Asian, African, and European origins. We identified 45 parsimoniously informative sites and multiple substitutions per site within the concatenated (945 bp) nucleotide dataset, implying that probabilistic phylogenetic analysis methods are needed. High diversity and signatures of uniquely shared haplotypes with diverse localities combined with the trade data suggested multiple incursions and introduction origins in Brazil. Increasing agricultural/horticultural trade activities between the Old and New Worlds represents a significant biosecurity risk factor. Identifying pest origins will enable resistance profiling that reflects countries of origin to be included when developing a resistance management strategy, while identifying incursion pathways will improve biosecurity protocols and risk analysis at biosecurity hotspots including national ports.

  11. Multicopper oxidase-1 is required for iron homeostasis in Malpighian tubules of Helicoverpa armigera.

    PubMed

    Liu, Xiaoming; Sun, Chengxian; Liu, Xiaoguang; Yin, Xinming; Wang, Baohai; Du, Mengfang; An, Shiheng

    2015-10-06

    Multicopper oxidases (MCOs) are enzymes that contain 10 conserved histidine residues and 1 cysteine residue. MCO1 has been extensively investigated in the midgut because this MCO is implicated in ascorbate oxidation, iron homeostasis and immune responses. However, information regarding the action of MCO1 in Malpighian tubules is limited. In this study, Helicoverpa armigera was used as a model to investigate the function of MCO1 in Malpighian tubules. Sequence analysis results revealed that HaMCO1 exhibits typical MCO characteristics, with 10 histidine and 1 cysteine residues for copper ion binding. HaMCO1 was also found to be highly abundant in Malpighian tubules. Temporal expression patterns indicated that HaMCO1 is mainly expressed during larval molting stages. Hormone treatments [the molting hormone 20-hydroxyecdysone (20E) and juvenile hormone (JH)] revealed that 20E inhibits HaMCO1 transcript expression via its heterodimer receptor, which consists of ecdysone receptor (EcR) and ultraspiracle (USP), and that JH counteracts the action of 20E to activate HaMCO1 transcript expression via its intracellular receptor methoprene-tolerant (Met). HaMCO1 knockdown caused a significant decrease in iron accumulation and also significantly reduced transferrin and ferritin transcript expression. Therefore, HaMCO1 is coordinately regulated by 20E and JH and is required for iron homeostasis in Malpighian tubules.

  12. Defensive role of tomato polyphenol oxidases against cotton bollworm (Helicoverpa armigera) and beet armyworm (Spodoptera exigua).

    PubMed

    Bhonwong, Anongnut; Stout, Michael J; Attajarusit, Jutharat; Tantasawat, Piyada

    2009-01-01

    Tomato (Solanum lycopersicum) polyphenol oxidases (PPOs), enzymes that oxidize phenolics to quinones, have been implicated in plant resistance to insects. The role of PPO in resistance to cotton bollworm [Helicoverpa armigera (Hübner)] and beet armyworm [Spodoptera exigua (Hübner)] (Lepidoptera: Noctuidae) was evaluated. Consumption, weight gains, and mortality of larvae feeding on foliage of transgenic tomato lines overexpressing PPO (OP lines) and of larvae feeding on foliage of transgenic tomato lines with suppressed PPO (SP lines) were compared with consumption, weight gains, and mortality of larvae feeding on non-transformed (NT) plants. Increases in foliage consumption and weight gains were observed for cotton bollworms feeding on leaves of SP plants compared to NT and OP plants. PPO activity was negatively correlated with both weight gains and foliar consumption of cotton bollworm, substantiating the defensive role of PPO against this insect. Similarly, beet armyworm consumed less foliage (both young and old leaves) from OP plants than SP plants. Larvae feeding on OP leaves generally exhibited lower weight gains than those feeding on SP leaves. These results indicate that tomato PPO plays a role in resistance to both cotton bollworm and beet armyworm.

  13. Mis-splicing of the ABCC2 gene linked with Bt toxin resistance in Helicoverpa armigera

    PubMed Central

    Xiao, Yutao; Zhang, Tao; Liu, Chenxi; Heckel, David G.; Li, Xianchun; Tabashnik, Bruce E.; Wu, Kongming

    2014-01-01

    Toxins from the bacterium Bacillus thuringiensis (Bt) are used widely for insect control in sprays and transgenic plants, but their efficacy is reduced when pests evolve resistance. Previous work showed that mutations in a gene encoding the transporter protein ABCC2 are linked with resistance to Bt toxins Cry1Ab, Cry1Ac or both in four species of Lepidoptera. Here we compared the ABCC2 gene of Helicoverpa armigera (HaABCC2) between susceptible strains and a laboratory-selected strain with >1,000-fold resistance to Cry1Ac relative its susceptible parent strain. We discovered a 73-base pair (bp) insertion in the cDNA of the resistant strain that generates a premature stop codon expected to yield a truncated ABCC2 protein. Sequencing of genomic DNA revealed that this insertion is an intron that is not spliced out because of a 6-bp deletion at its splicing site. Analysis of progeny from crosses revealed tight genetic linkage between HaABCC2 and resistance to Cry1Ac. These results provide the first evidence that mis-splicing of a gene encoding an ABCC2 protein confers resistance to a Bt toxin. PMID:25154974

  14. A cathepsin L-like proteinase is involved in moulting and metamorphosis in Helicoverpa armigera.

    PubMed

    Wang, L-F; Chai, L-Q; He, H-J; Wang, Q; Wang, J-X; Zhao, X-F

    2010-02-01

    Holometabolous insects undergo larval moulting and metamorphosis within their life cycle. A cDNA encoding the cathepsin L-like proteinase Ha-cathL has been cloned from Helicoverpa armigera. It has a sequence of 1826 bp and encodes a 550-residue protein with a molecular mass of 63 kDa. Northern blot analysis indicated that Ha-cathL is specifically expressed in haemocytes, with increased expression during larval moulting and metamorphosis. In vivo experimentation revealed that Ha-cathL is up-regulated by 20-hydroxyecdysone. Meanwhile, in situ hybridization and immunocytochemistry revealed that Ha-cathL mRNA is mainly expressed in granulocytes and plasmatocytes. Knock down of cathepsin L by RNA interference results in larvae death before pupation or the formation of a chimeric pupa containing a larval head and thorax, abnormal wings and the pupal abdomen. The reason for this is that the affected haemocytes cannot become granulated, and therefore cannot participate in fat body remodelling and wing development. These facts suggest that Ha-cathL is involved in larval moulting and metamorphosis by participating in the functioning of haemocytes.

  15. Multicopper oxidase-1 is required for iron homeostasis in Malpighian tubules of Helicoverpa armigera

    PubMed Central

    Liu, Xiaoming; Sun, Chengxian; Liu, Xiaoguang; Yin, Xinming; Wang, Baohai; Du, Mengfang; An, Shiheng

    2015-01-01

    Multicopper oxidases (MCOs) are enzymes that contain 10 conserved histidine residues and 1 cysteine residue. MCO1 has been extensively investigated in the midgut because this MCO is implicated in ascorbate oxidation, iron homeostasis and immune responses. However, information regarding the action of MCO1 in Malpighian tubules is limited. In this study, Helicoverpa armigera was used as a model to investigate the function of MCO1 in Malpighian tubules. Sequence analysis results revealed that HaMCO1 exhibits typical MCO characteristics, with 10 histidine and 1 cysteine residues for copper ion binding. HaMCO1 was also found to be highly abundant in Malpighian tubules. Temporal expression patterns indicated that HaMCO1 is mainly expressed during larval molting stages. Hormone treatments [the molting hormone 20-hydroxyecdysone (20E) and juvenile hormone (JH)] revealed that 20E inhibits HaMCO1 transcript expression via its heterodimer receptor, which consists of ecdysone receptor (EcR) and ultraspiracle (USP), and that JH counteracts the action of 20E to activate HaMCO1 transcript expression via its intracellular receptor methoprene-tolerant (Met). HaMCO1 knockdown caused a significant decrease in iron accumulation and also significantly reduced transferrin and ferritin transcript expression. Therefore, HaMCO1 is coordinately regulated by 20E and JH and is required for iron homeostasis in Malpighian tubules. PMID:26437857

  16. Influence of CO2 and Temperature on Metabolism and Development of Helicoverpa armigera (Noctuidae: Lepidoptera).

    PubMed

    Akbar, S Md; Pavani, T; Nagaraja, T; Sharma, H C

    2016-02-01

    Climate change will have a major bearing on survival and development of insects as a result of increase in CO2 and temperature. Therefore, we studied the direct effects of CO2 and temperature on larval development and metabolism in cotton bollworm, Helicoverpa armigera (Hübner). The larvae were reared under a range of CO2 (350, 550, and 750 ppm) and temperature (15, 25, 35, and 45°C) regimes on artificial diet. Elevated CO2 negatively affected the larval survival, larval weight, larval period, pupation, and adult emergence, but showed a positive effect on pupal weight, pupal period, and fecundity. Increase in temperature exhibited a negative effect on larval survival, larval period, pupal weights, and pupal period, but a positive effect on larval growth. Pupation and adult emergence were optimum at 25°C. Elevated CO2 and temperature increased food consumption and metabolism of larvae by enhancing the activity of midgut proteases, carbohydrases (amylase and cellulase), and mitochondrial enzymes and therefore may cause more damage to crop production. Elevated CO2 and global warming will affect insect growth and development, which will change the interactions between the insect pests and their crop hosts. Therefore, there is need to gain an understanding of these interactions to develop strategies for mitigating the effects of climate change.

  17. Current status of insecticide resistance in Helicoverpa armigera after 15 years of Bt cotton planting in China.

    PubMed

    Yang, Yihua; Li, Yapeng; Wu, Yidong

    2013-02-01

    Insecticide resistance was an important factor responsible for outbreaks of Helicoverpa armigera (Hübner) in China in the early 1990s. Bt cotton has been adopted in China since 1997, and has resulted in a reduction of insecticide use for H. armigera control. After 15 yr of Bt cotton planting, in 2011 we surveyed resistance to fenvalerate, phoxim, and emamectin benzoate in 16 field populations of H. armigera collected from major cotton production areas of China. Fourteen populations from northern China showed very strong resistance to fenvalerate (from 43- to 830-fold) and low levels of resistance to phoxim (3.0- to 8.9-fold) when compared with the susceptible SCD strain of H. armigera, whereas two populations from northwestern China showed low levels of resistance to fenvalerate (3.0- and 10-fold) and no resistance to phoxim (0.7- and 0.9-fold). Synergist bioassays demonstrated that oxidase-based detoxification was involved in fenvalerate resistance and esterase-based detoxification in phoxim resistance in the resistant field populations. In comparison with the resistance in field populations before Bt cotton adoption, we observed a maintenance of high levels of fenvalerate resistance, but a reversion of phoxim resistance from high levels to low levels in the field populations of H. armigera from northern China. All 16 field populations from both northern China and northwestern China were susceptible to emamectin benzoate (with about two-fold variations in LD50s among populations), but the SCD strain has an inherent tolerance of 11-fold compared with the most susceptible field population (Xiajin-1). Emamectin benzoate is not cross resistant to fenvalerate and may provide an alternative option for H. armigera control in China, if the efficacy of Bt cotton is compromised by Bt resistance in the field.

  18. Comparison of Leg Regeneration Potency Between Holometabolous Helicoverpa armigera (Lepidoptera: Noctuidae) and Hemimetabolous Locusta migratoria manilensis (Orthoptera: Acrididae).

    PubMed

    Yang, Qingpo; Li, Zhen; Li, Hui; Li, Yanrong; Yang, Yuhui; Zhang, Qingwen; Liu, Xiaoxia

    2016-09-11

    After injury many insects could regenerate lost limb. In this study, Helicoverpa armigera Hubner and Locusta migratoria manilensis (Meyen, 1835) were chosen to compare the regeneration potency of holometabolous and hemimetabolous insects. We employed the classical approach of surgical excision to verify the regeneration ability and to investigate the factors that affect the extent of regeneration. The results found that H. armigera could regenerate intact legs when the larval legs were excised at the first and second instar and that legs of adult H. armigera had a close relationship with their larval counterparts. However, the adult legs became malformed or disappeared when excised at other older instars. For the L. migratoria, we found the legs have weak partial regeneration ability when amputation was conducted at the joint of two segments. The regeneration potency might be stronger the more proximal the operation. Regeneration process had a negative impact on the larval development. This is the first report of complete leg regeneration capacity having a strong correlation with the instar but not with the position where amputation occurred for H. armigera, while for the L. migratoria, partial regenerative ability had a close relationship with the position where amputation occurred but not with instars.

  19. Comparison of Leg Regeneration Potency Between Holometabolous Helicoverpa armigera (Lepidoptera: Noctuidae) and Hemimetabolous Locusta migratoria manilensis (Orthoptera: Acrididae).

    PubMed

    Yang, Qingpo; Li, Zhen; Li, Hui; Li, Yanrong; Yang, Yuhui; Zhang, Qingwen; Liu, Xiaoxia

    2016-12-01

    After injury many insects could regenerate lost limb. In this study, Helicoverpa armigera Hubner and Locusta migratoria manilensis (Meyen, 1835) were chosen to compare the regeneration potency of holometabolous and hemimetabolous insects. We employed the classical approach of surgical excision to verify the regeneration ability and to investigate the factors that affect the extent of regeneration. The results found that H. armigera could regenerate intact legs when the larval legs were excised at the first and second instar and that legs of adult H. armigera had a close relationship with their larval counterparts. However, the adult legs became malformed or disappeared when excised at other older instars. For the L. migratoria, we found the legs have weak partial regeneration ability when amputation was conducted at the joint of two segments. The regeneration potency might be stronger the more proximal the operation. Regeneration process had a negative impact on the larval development. This is the first report of complete leg regeneration capacity having a strong correlation with the instar but not with the position where amputation occurred for H. armigera, while for the L. migratoria, partial regenerative ability had a close relationship with the position where amputation occurred but not with instars.

  20. The efficacy of Beauveria bassiana, jasmonic acid and chlorantraniliprole on larval populations of Helicoverpa armigera in chickpea crop ecosystems.

    PubMed

    Younas, Aneela; Wakil, Waqas; Khan, Zaeema; Shaaban, Muhammad; Prager, Sean Michael

    2017-02-01

    A robust integrated pest management (IPM) programme is needed to reduce the use of insecticides in controlling Helicoverpa armigera. Therefore, a 2 year field study was conducted to evaluate the use of alternative control measures (biochemical use) for H. armigera relative to exclusively using chemical insecticides. The entomopathogenic fungus Beauveria bassiana, jasmonic acid and the insecticide chlorantraniliprole were each applied twice during the chickpea growing season. All three applied materials (either alone or combined) significantly (P ≤ 0.05) reduced the larval population of H. armigera and pod infestation. Effects increased with time, and the maximum difference was observed 7 days after the second application in each year. The lowest numbers of larvae per plant and pod infestation were in the B. bassiana 3.21 × 10(6) + chlorantraniliprole treatment in both 2009/2010 and 2010/2011 year. The reduction in the larval population and pod infestation increased chickpea yield and the highest yield in both seasons, and the maximum yield was obtained in the B. bassiana 3.21 × 10(6) + chlorantraniliprole treatment. The populations of natural enemies were highest in the jasmonic acid treatment. The results suggest that B. bassiana, jasmonic acid and chlorantraniliprole may be useful components for the H. armigera IPM strategy. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  1. Comparisons of contact chemoreception and food acceptance by larvae of polyphagous Helicoverpa armigera and oligophagous Bombyx mori.

    PubMed

    Zhang, Hui-Jie; Faucher, Cécile P; Anderson, Alisha; Berna, Amalia Z; Trowell, Stephen; Chen, Quan-Mei; Xia, Qing-You; Chyb, Sylwester

    2013-08-01

    We compared food choice and the initial response to deterrent treated diet between fifth instars of Helicoverpa armigera, a polyphagous generalist pest, and Bombyx mori, an oligophagous specialist beneficial. Bombyx mori was more behaviorally sensitive to salicin than to caffeine. The relative sensitivities were reversed for H. armigera, which was tolerant to the highest levels of salicin found in natural sources but sensitive to caffeine. A single gustatory receptor neuron (GRN) in the medial styloconic sensillum of B. mori was highly sensitive to salicin and caffeine. The styloconic sensilla of H. armigera did not respond consistently to either of the bitter compounds. Phagostimulants also were tested. Myo-inositol and sucrose were detected specifically by two GRNs located in B. mori lateral styloconic sensillum, whereas, in H. armigera, sucrose was sensed by a GRN in the lateral sensillum, and myo-inositol by a GRN in the medial sensillum. Myo-inositol responsiveness in both species occurred at or below 10(-3) mM, which is far below the naturally occurring concentration of 1 mM in plants. Larval responses to specific plant secondary compounds appear to have complex determinants that may include host range, metabolic capacity, and gustatory repertoire.

  2. Mitochondrial P-glycoprotein ATPase contributes to insecticide resistance in the cotton bollworm, Helicoverpa armigera (Noctuidae: Lepidoptera).

    PubMed

    Akbar, S Md; Aurade, Ravindra M; Sharma, H C; Sreeramulu, K

    2014-09-01

    Cotton bollworm, Helicoverpa armigera, is one of the most damaging polyphagous pests worldwide, which has developed high levels of resistance to commonly applied insecticides. Mitochondrial P-glycoprotein (Pgp) was detected in the insecticide-resistant strain of H. armigera using C219 antibodies, and its possible role was demonstrated in the efflux of xenobiotic compounds using spectrofluorometer. The TMR accumulated in mitochondria in the absence of ATP, and effluxed out in presence of ATP; the process of efflux was inhibited in the presence of ortho-vandate, an inhibitor of Pgp, in insecticide-resistant larvae of H. armigera. The mitochondria isolated from insecticide-resistant larvae were resistant to insecticide-induced inhibition of oxygen consumption and cytochrome c release. Membrane potential decreased in a dose-dependent manner in the presence of higher concentration of insecticides (>50 µM) in mitochondria of insecticide-resistant larvae. In conclusion, mitochondrial Pgp ATPase detected in the insecticide-resistant larvae influenced the efflux of xenobiotic compounds. Pgp might be involved in protecting the mitochondrial DNA and the components of the electron transport chain from damage due to insecticides, and contributing to the resistance to the deleterious effects of insecticides on the growth of insecticide-resistant H. armigera larvae.

  3. Biological activities of Solanum pseudocapsicum (Solanaceae) against cotton bollworm, Helicoverpa armigera Hübner and armyworm, Spodoptera litura Fabricius (Lepidotera: Noctuidae)

    PubMed Central

    Jeyasankar, Alagarmalai; Premalatha, Selvaraj; Elumalai, Kuppusamy

    2012-01-01

    Objective To evaluate the antifeedant, insecticidal and growth inhibition activities of Solanum pseudocapsicum (S. pseudocapsicum) seed extracts against Spodoptera litura (S. litura) and Helicoverpa armigera (H. armigera). Methods Hexane, diethyl ether, dichloromethane and ethyl acetate seed extracts were prepared and tested for antifeedant, insecticidal and growth inhibitory activities against fourth instar larvae of S. litura and H. armigera. Results Ethyl acetate extract showed promising antifeedant and insecticidal activities against S. litura and H. armigera. Percentage of deformed larvae, pupae and adults were maximum in treatment of ethyl acetate extract. Percentage of successful adult emergence was deteriorated by seeds on extract treated larvae. Conclusions Ethyl acetate extracts of S. pseudocapsicum, showed higher efficiency of antifeedant, insecticidal and growth inhibition activities. Hence, it can be used to controll agricultural insect pests, S. litura and H. armigera. PMID:23593579

  4. Determinant Factors in the Production of a Co-Occluded Binary Mixture of Helicoverpa armigera Alphabaculovirus (HearNPV) Genotypes with Desirable Insecticidal Characteristics.

    PubMed

    Arrizubieta, Maite; Simón, Oihane; Williams, Trevor; Caballero, Primitivo

    2016-01-01

    A co-occluded binary mixture of Helicoverpa armigera nucleopolyhedrovirus genotypes HearSP1B and HearLB6 at a 1:1 ratio (HearSP1B+HearLB6) was selected for the development of a virus-based biological insecticide, which requires an efficient large-scale production system. In vivo production systems require optimization studies in each host-virus pathosystem. In the present study, the effects of larval instar, rearing density, timing of inoculation, inoculum concentration and temperature on the production of HearSP1B+HearLB6 in its homologous host were evaluated. The high prevalence of cannibalism in infected larvae (40-87%) indicated that insects require individual rearing to avoid major losses in OB production. The OB production of recently molted fifth instars (7.0 x 109 OBs/larva), combined with a high prevalence of mortality (85.7%), resulted in the highest overall OB yield (6.0 x 1011 OBs/100 inoculated larvae), compared to those of third or fourth instars. However, as inoculum concentration did not influence final OB yield, the lowest concentration, LC80 (5.5 x 106 OBs/ml), was selected. Incubation temperature did not significantly influence OB yield, although larvae maintained at 30°C died 13 and 34 hours earlier than those incubated at 26°C and 23°C, respectively. We conclude that the efficient production of HearSP1B+HearLB6 OBs involves inoculation of recently molted fifth instars with a LC80 concentration of OBs followed by individual rearing at 30°C.

  5. Determinant Factors in the Production of a Co-Occluded Binary Mixture of Helicoverpa armigera Alphabaculovirus (HearNPV) Genotypes with Desirable Insecticidal Characteristics

    PubMed Central

    Arrizubieta, Maite; Simón, Oihane; Williams, Trevor; Caballero, Primitivo

    2016-01-01

    A co-occluded binary mixture of Helicoverpa armigera nucleopolyhedrovirus genotypes HearSP1B and HearLB6 at a 1:1 ratio (HearSP1B+HearLB6) was selected for the development of a virus-based biological insecticide, which requires an efficient large-scale production system. In vivo production systems require optimization studies in each host-virus pathosystem. In the present study, the effects of larval instar, rearing density, timing of inoculation, inoculum concentration and temperature on the production of HearSP1B+HearLB6 in its homologous host were evaluated. The high prevalence of cannibalism in infected larvae (40–87%) indicated that insects require individual rearing to avoid major losses in OB production. The OB production of recently molted fifth instars (7.0 x 109 OBs/larva), combined with a high prevalence of mortality (85.7%), resulted in the highest overall OB yield (6.0 x 1011 OBs/100 inoculated larvae), compared to those of third or fourth instars. However, as inoculum concentration did not influence final OB yield, the lowest concentration, LC80 (5.5 x 106 OBs/ml), was selected. Incubation temperature did not significantly influence OB yield, although larvae maintained at 30°C died 13 and 34 hours earlier than those incubated at 26°C and 23°C, respectively. We conclude that the efficient production of HearSP1B+HearLB6 OBs involves inoculation of recently molted fifth instars with a LC80 concentration of OBs followed by individual rearing at 30°C. PMID:27732657

  6. Antibiotics influence the toxicity of the delta endotoxins of Bacillus thuringiensis towards the cotton bollworm, Helicoverpa armigera.

    PubMed

    Paramasiva, Inakarla; Sharma, Hari C; Krishnayya, Pulipaka Venkata

    2014-07-24

    The cotton bollworm, Helicoverpa armigera is one of the most important crop pests worldwide. It has developed high levels of resistance to synthetic insecticides, and hence, Bacillus thuringiensis (Bt) formulations are used as a safer pesticide and the Bt genes have been deployed in transgenic crops for controlling this pest. There is an apprehension that H. armigera might develop resistance to transgenic crops in future. Therefore, we studied the role of gut microbes by eliminating them with antibiotics in H. armigera larvae on the toxicity of Bt toxins against this pest. Commercial formulation of Bt (Biolep®) and the pure Cry1Ab and Cry1Ac toxin proteins were evaluated at ED50, LC50, and LC90 dosages against the H. armigera larvae with and without antibiotics (which removed the gut microbes). Lowest H. armigera larval mortality due to Bt formulation and the Bt toxins Cry1Ab and Cry1Ac was recorded in insects reared on diets with 250 and 500 μg ml-1 diet of each of the four antibiotics (gentamicin, penicillin, rifampicin, and streptomycin), while the highest larval mortality was recorded in insects reared on diets without the antibiotics. Mortality of H. armigera larvae fed on diets with Bt formulation and the δ-endotoxins Cry1Ab and Cry1Ac was inversely proportional to the concentration of antibiotics in the artificial diet. Nearly 30% reduction in larval mortality was observed in H. armigera larvae from F1 to F3 generation when the larvae were reared on diets without antibiotics (with gut microbes) and fed on 0.15% Bt or 12 μg Cry1Ab or Cry1Ac ml-1 diet, indicating development of resistance to Bt in the presence of gut microflora. However, there were no differences in larval mortality due to Bt, Cry1Ab or Cry1Ac across generations in insects when they were reared on diets with 250 μg of each antibiotic ml-1 diet (without gut microflora). The results suggested that antibiotics which eliminated gut microflora influenced the toxicity of Bt towards H. armigera

  7. Antibiotics influence the toxicity of the delta endotoxins of Bacillus thuringiensis towards the cotton bollworm, Helicoverpa armigera

    PubMed Central

    2014-01-01

    Background The cotton bollworm, Helicoverpa armigera is one of the most important crop pests worldwide. It has developed high levels of resistance to synthetic insecticides, and hence, Bacillus thuringiensis (Bt) formulations are used as a safer pesticide and the Bt genes have been deployed in transgenic crops for controlling this pest. There is an apprehension that H. armigera might develop resistance to transgenic crops in future. Therefore, we studied the role of gut microbes by eliminating them with antibiotics in H. armigera larvae on the toxicity of Bt toxins against this pest. Results Commercial formulation of Bt (Biolep®) and the pure Cry1Ab and Cry1Ac toxin proteins were evaluated at ED50, LC50, and LC90 dosages against the H. armigera larvae with and without antibiotics (which removed the gut microbes). Lowest H. armigera larval mortality due to Bt formulation and the Bt toxins Cry1Ab and Cry1Ac was recorded in insects reared on diets with 250 and 500 μg ml−1 diet of each of the four antibiotics (gentamicin, penicillin, rifampicin, and streptomycin), while the highest larval mortality was recorded in insects reared on diets without the antibiotics. Mortality of H. armigera larvae fed on diets with Bt formulation and the δ-endotoxins Cry1Ab and Cry1Ac was inversely proportional to the concentration of antibiotics in the artificial diet. Nearly 30% reduction in larval mortality was observed in H. armigera larvae from F1 to F3 generation when the larvae were reared on diets without antibiotics (with gut microbes) and fed on 0.15% Bt or 12 μg Cry1Ab or Cry1Ac ml−1 diet, indicating development of resistance to Bt in the presence of gut microflora. However, there were no differences in larval mortality due to Bt, Cry1Ab or Cry1Ac across generations in insects when they were reared on diets with 250 μg of each antibiotic ml−1 diet (without gut microflora). Conclusions The results suggested that antibiotics which eliminated gut microflora

  8. Differential responses of Helicoverpa armigera C-type immunlectin genes to the endoparasitoid Campoletis chlorideae.

    PubMed

    Wang, Xiong-Ya; Bai, Su-Fen; Li, Xin; An, Shi-Heng; Yin, Xin-Ming; Li, Xian-Chun

    2017-03-01

    The C-type lectins mediate nonself recognition in insects. The previous studies focused on host immunlectin response to bacterial infection; however, the molecular basis of immunlectin reactions to endoparasitoids has not been elucidated. The present study investigated the effect of parasitization by Campoletis chlorideae on hemagglutination activity (HA; defined as the ability of lectin to agglutinate erythrocytes or other cells), and transcriptional expression of C-type immunlectin genes in the larval host, Helicoverpa armigera. Parasitization induced four- to eightfold higher HA in the parasitized larvae, compared to nonparasitized larvae at days 2 and 6 postparasitization (PP), however inhibited HA at other days PP. Eight C-type lectins were differentially expressed in different host developmental stages, from feeding to wandering stage. The mRNA levels of HaCTL1, HaCTL3, HaCTL4, and HaCTL5 were upregulated and HaCTL2 and HaCTL7 were downregulated. Tissue analysis showed that HaCTLs were mainly expressed in fat body or hemocytes, while HaCTL5 was highly expressed in testes. The effects of parasitization on the lectin expression patterns differed. Lectins except HaCTL6 or HaCTL5 were significantly down- or upregulated in parasitized larvae at day 4 or 6 PP compared with that of nonparasitized larvae. We infer from our results that C-type immunlectins are involved in host-parasitoid interactions, and parasitization alter host immunlectin levels both in inhibiting and promoting host immune defenses to endoparasitoids. These immunlectin genes indicated an altered physiological status of the host insect, depending on developmental stage, tissue, and parasitization.

  9. Heterologous Expression and Biochemical Characterisation of Fourteen Esterases from Helicoverpa armigera

    PubMed Central

    Li, Yongqiang; Coppin, Chris W.; Devonshire, Alan L.; Scott, Colin; East, Peter; Russell, Robyn J.; Oakeshott, John G.

    2013-01-01

    Esterases have recurrently been implicated in insecticide resistance in Helicoverpa armigera but little is known about the underlying molecular mechanisms. We used a baculovirus system to express 14 of 30 full-length esterase genes so far identified from midgut cDNA libraries of this species. All 14 produced esterase isozymes after native PAGE and the isozymes for seven of them migrated to two regions of the gel previously associated with both organophosphate and pyrethroid resistance in various strains. Thirteen of the enzymes obtained in sufficient yield for further analysis all showed tight binding to organophosphates and low but measurable organophosphate hydrolase activity. However there was no clear difference in activity between the isozymes from regions associated with resistance and those from elsewhere in the zymogram, or between eight of the isozymes from a phylogenetic clade previously associated with resistance in proteomic and quantitative rtPCR experiments and five others not so associated. By contrast, the enzymes differed markedly in their activities against nine pyrethroid isomers and the enzymes with highest activity for the most insecticidal isomers were from regions of the gel and, in some cases, the phylogeny that had previously been associated with pyrethroid resistance. Phospholipase treatment confirmed predictions from sequence analysis that three of the isozymes were GPI anchored. This unusual feature among carboxylesterases has previously been suggested to underpin an association that some authors have noted between esterases and resistance to the Cry1Ac toxin from Bacillus thuringiensis. However these three isozymes did not migrate to the zymogram region previously associated with Cry1Ac resistance. PMID:23799064

  10. Vip3Aa tolerance response of Helicoverpa armigera populations from a Cry1Ac cotton planting region.

    PubMed

    An, Jingjie; Gao, Yulin; Wu, Kongming; Gould, Fred; Gao, Jianhua; Shen, Zhicheng; Lei, Chaoliang

    2010-12-01

    Transgenic cotton, Gossypium hirsutum L., that expresses the Bacillus thuringiensis (Bt) Cry1Ac toxin, holds great promise in controlling target insect pests. Evolution of resistance by target pests is the primary threat to the continued efficacy of Bt cotton. To thwart pest resistance evolution, a transgenic cotton culitvar that produces two different Bt toxins, cry1Ac and vip3A genes, was proposed as a successor of cry1Ac cotton. This article reports on levels of Vip3Aa tolerance in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) populations from the Cry1Ac cotton planting region in China based on bioassays of the F1 generation of isofemale lines. In total, 80 isofemale families of H. armigera from Xiajin county of Shandong Province (an intensive Bt cotton planting area) and 93 families from Anci county of Hebei Province (a multiple-crop system including corn [Zea mays L.] , soybean [Glycine max (L.) Merr.], peanut (Arachis hypogaea L.), and Bt cotton) were screened with a discriminating concentration of both Cry1Ac- and Vip3A-containing diets in 2009. From data on the relative average development rates and percentage of larval weight inhibition of F1 full-sib families tested simultaneously on Cry1Ac and Vip3Aa, results indicate that responses to Cry1Ac and Vip3Aa were not genetically correlated in field population ofH. armigera. This indicates that the threat of cross-resistance between Cry1Ac and Vip3A is low in field populations of H. armigera. Thus, the introduction of Vip3Aa/Cry1Ac-producing lines could delay resistance evolution in H. armigera in Bt cotton planting area of China.

  11. Elimination of Gut Microbes with Antibiotics Confers Resistance to Bacillus thuringiensis Toxin Proteins in Helicoverpa armigera (Hubner).

    PubMed

    Visweshwar, R; Sharma, H C; Akbar, S M D; Sreeramulu, K

    2015-12-01

    Helicoverpa armigera is one of the most important pests worldwide. Transgenic crops with toxin genes from Bacillus thuringiensis (Bt) have been deployed on a large scale to control this pest. The insecticidal activity of Bt is probably influenced by the insect midgut microbes, which vary across crop hosts and locations. Therefore, we examined the role of gut microbes in pathogenicity of Bt toxins in the H. armigera. Antibiotic cocktail was used for the complete elimination of the H. armigera gut microbes. Activated Cry1Ac, Bt formulation, and transgenic cotton resulted in larval weight loss and increase in mortality, but pretreatment of larvae with antibiotic cocktail significantly decreased larval mortality and increased the larval weight gain. Activated Cry1Ac and Bt formulation inhibited the activity of proteases in midgut of H. armigera larvae but showed no such effect in the larvae pretreated with antibiotic cocktail. Five protease bands in activated Cry1Ac and two in Bt formulation-treated larvae were inhibited but no such effect in the larvae pretreated with antibiotic cocktail. Cry1Ac protein was detected in Bt/Cry1Ac protoxin-fed larval gut extract in the absence of antibiotic cocktail, but fewer in larvae pretreated with antibiotic cocktail. The activity of antioxidant enzymes and aminopeptidases increased in larvae fed on Bt toxin, but there was no significant increase in antioxidant enzymes in larvae reared on toxin protein in combination with antibiotic cocktail. The results suggest that gut microbes exercise a significant influence on the toxicity of Cry1Ac and Bt formulation in H. armigera larvae. The implications of these results have been discussed in relation to development of insect resistance to Bt transgenic crops deployed for pest management.

  12. Elevated CO2 Reduces the Resistance and Tolerance of Tomato Plants to Helicoverpa armigera by Suppressing the JA Signaling Pathway

    PubMed Central

    Ren, Qin; Zhu-Salzman, Keyan; Kang, Le; Wang, Chenzhu; Li, Chuanyou; Ge, Feng

    2012-01-01

    Both resistance and tolerance, which are two strategies that plants use to limit biotic stress, are affected by the abiotic environment including atmospheric CO2 levels. We tested the hypothesis that elevated CO2 would reduce resistance (i.e., the ability to prevent damage) but enhance tolerance (i.e., the ability to regrow and compensate for damage after the damage has occurred) of tomato plants to the cotton bollworm, Helicoverpa armigera. The results showed that elevated CO2 reduced resistance by decreasing the jasmonic acid (JA) level and activities of lipoxygenase, proteinase inhibitors, and polyphenol oxidase in wild-type (WT) plants infested with H. armigera. Consequently, the activities of total protease, trypsin-like enzymes, and weak and active alkaline trypsin-like enzymes increased in the midgut of H. armigera when fed on WT plants grown under elevated CO2. Unexpectedly, the tolerance of the WT to H. armigera (in terms of photosynthetic rate, activity of sucrose phosphate synthases, flower number, and plant biomass and height) was also reduced by elevated CO2. Under ambient CO2, the expression of resistance and tolerance to H. armigera was much greater in wild type than in spr2 (a JA-deficient genotype) plants, but elevated CO2 reduced these differences of the resistance and tolerance between WT and spr2 plants. The results suggest that the JA signaling pathway contributes to both plant resistance and tolerance to herbivorous insects and that by suppressing the JA signaling pathway, elevated CO2 will simultaneously reduce the resistance and tolerance of tomato plants. PMID:22829948

  13. Diversity in gut microflora of Helicoverpa armigera populations from different regions in relation to biological activity of Bacillus thuringiensis δ-endotoxin Cry1Ac.

    PubMed

    Paramasiva, Inakarla; Shouche, Yogesh; Kulkarni, Girish Jayant; Krishnayya, Pulipaka Venkata; Akbar, Shaik Mohammed; Sharma, Hari Chand

    2014-12-01

    Transgenic crops expressing toxin proteins from Bacillus thuringiensis (Bt) have been deployed on a large scale for management of Helicoverpa armigera. Resistance to Bt toxins has been documented in several papers, and therefore, we examined the role of midgut microflora of H. armigera in its susceptibility to Bt toxins. The susceptibility of H. armigera to Bt toxin Cry1Ac was assessed using Log-dose-Probit analysis, and the microbial communities were identified by 16S rRNA sequencing. The H. armigera populations from nine locations harbored diverse microbial communities, and had some unique bacteria, suggesting a wide geographical variation in microbial community in the midgut of the pod borer larvae. Phylotypes belonging to 32 genera were identified in the H. armigera midgut in field populations from nine locations. Bacteria belonging to Enterobacteriaceae (Order Bacillales) were present in all the populations, and these may be the common members of the H. armigera larval midgut microflora. Presence and/or absence of certain species were linked to H. armigera susceptibility to Bt toxins, but there were no clear trends across locations. Variation in susceptibility of F1 neonates of H. armigera from different locations to the Bt toxin Cry1Ac was found to be 3.4-fold. These findings support the idea that insect migut microflora may influence the biological activity of Bt toxins.

  14. Silencing the HaAK gene by transgenic plant-mediated RNAi impairs larval growth of Helicoverpa armigera.

    PubMed

    Liu, Feng; Wang, Xiao-Dong; Zhao, Yi-Ying; Li, Yan-Jun; Liu, Yong-Chang; Sun, Jie

    2015-01-01

    Insect pests have caused noticeable economic losses in agriculture, and the heavy use of insecticide to control pests not only brings the threats of insecticide resistance but also causes the great pollution to foods and the environment. Transgenic plants producing double-stranded RNA (dsRNA) directed against insect genes have been is currently developed for protection against insect pests. In this study, we used this technology to silence the arginine kinase (AK) gene of Helicoverpa armigera (HaAK), encoding a phosphotransferase that plays a critical role in cellular energy metabolism in invertebrate. Transgenic Arabidopsis plants producing HaAK dsRNA were generated by Agrobacterium-mediated transformation. The maximal mortality rate of 55% was reached when H. armigera first-instar larvae were fed with transgenic plant leaves for 3 days, which was dramatically higher than the 18% mortality recorded in the control group. Moreover, the ingestion of transgenic plants significantly retarded larval growth, and the transcript levels of HaAK were also knocked down by up to 52%. The feeding bioassays further indicated that the inhibition efficiency was correlated with the integrity and concentration of the produced HaAK dsRNA in transgenic plants. These results strongly show that the resistance to H. armigera was improved in transgenic Arabidopsis plants, suggesting that the RNAi targeting of AK has the potential for the control of insect pests.

  15. Silencing the HaHR3 Gene by Transgenic Plant-mediated RNAi to Disrupt Helicoverpa armigera Development

    PubMed Central

    Xiong, Yehui; Zeng, Hongmei; Zhang, Yuliang; Xu, Dawei; Qiu, Dewen

    2013-01-01

    RNA interference (RNAi) caused by exogenous double-stranded RNA (dsRNA) has developed into a powerful technique in functional genomics, and to date it is widely used to down-regulate crucial physiology-related genes to control pest insects. A molt-regulating transcription factor gene, HaHR3, of cotton bollworm (Helicoverpa armigera) was selected as the target gene. Four different fragments covering the coding sequence (CDS) of HaHR3 were cloned into vector L4440 to express dsRNAs in Escherichia coli. The most effective silencing fragment was then cloned into a plant over-expression vector to express a hairpin RNA (hpRNA) in transgenic tobacco (Nicotiana tabacum). When H. armigera larvae were fed the E. coli or transgenic plants, the HaHR3 mRNA and protein levels dramatically decreased, resulting developmental deformity and larval lethality. The results demonstrate that both recombinant bacteria and transgenic plants could induce HaHR3 silence to disrupt H. armigera development, transgenic plant-mediated RNAi is emerging as a powerful approach for controlling insect pests. PMID:23630449

  16. Silencing the HaAK Gene by Transgenic Plant-Mediated RNAi Impairs Larval Growth of Helicoverpa armigera

    PubMed Central

    Liu, Feng; Wang, Xiao-Dong; Zhao, Yi-Ying; Li, Yan-Jun; Liu, Yong-Chang; Sun, Jie

    2015-01-01

    Insect pests have caused noticeable economic losses in agriculture, and the heavy use of insecticide to control pests not only brings the threats of insecticide resistance but also causes the great pollution to foods and the environment. Transgenic plants producing double-stranded RNA (dsRNA) directed against insect genes have been is currently developed for protection against insect pests. In this study, we used this technology to silence the arginine kinase (AK) gene of Helicoverpa armigera (HaAK), encoding a phosphotransferase that plays a critical role in cellular energy metabolism in invertebrate. Transgenic Arabidopsis plants producing HaAK dsRNA were generated by Agrobacterium-mediated transformation. The maximal mortality rate of 55% was reached when H. armigera first-instar larvae were fed with transgenic plant leaves for 3 days, which was dramatically higher than the 18% mortality recorded in the control group. Moreover, the ingestion of transgenic plants significantly retarded larval growth, and the transcript levels of HaAK were also knocked down by up to 52%. The feeding bioassays further indicated that the inhibition efficiency was correlated with the integrity and concentration of the produced HaAK dsRNA in transgenic plants. These results strongly show that the resistance to H. armigera was improved in transgenic Arabidopsis plants, suggesting that the RNAi targeting of AK has the potential for the control of insect pests. PMID:25552931

  17. Digestive proteolytic and amylolytic activities and feeding responses of Helicoverpa armigera (Lepidoptera: Noctuidae) on different host plants.

    PubMed

    Hemati, S A; Naseri, B; Ganbalani, G Nouri; Dastjerdi, H Rafiee; Golizadeh, A

    2012-08-01

    Digestive proteolytic and amylolytic activities and feeding responses of fifth instar larvae of Helicoverpa armigera (Hübner) on different host plants including chickpea (cultivars Arman, Hashem, Azad, and Binivich), common bean (cultivar Khomein), white kidney bean (cultivar Dehghan), red kidney bean (cultivar Goli), cowpea (cultivar Mashhad), tomato (cultivar Meshkin), and potato (cultivars Agria and Satina) were studied under laboratory conditions (25 +/- 1 degrees C, 65 +/- 5% RH and a photoperiod of 16:8 [L:D] h). Our results showed that the highest protease activity in optimal pH was on cultivar Dehghan (8.717 U/mg) and lowest one was on Meshkin (3.338 U/mg). In addition, the highest amylase activity in optimal pH was on cultivar Dehghan (0.340 mU/mg) and lowest was on Meshkin (0.088 mU/mg). The larval weight of fifth instar H. armigera showed significant difference, being heaviest on Binivich (125.290 +/- 5.050 mg) and lightest on Meshkin (22.773 +/- 0.575 mg). Furthermore, the highest and lowest values of food consumed were on Goli (362.800 +/- 27.500 mg) and Satina (51.280 +/- 4.500 mg), respectively. In addition, the lowest values of prepupal and pupal weight were on Meshkin (32.413 +/- 0.980 and 41.820 +/- 1.270 mg, respectively). The results indicated that tomato (Meshkin) was unsuitable host for feeding fifth instar larvae of H. armigera.

  18. Inheritance and stability of pyrethroid resistance in the cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) in Central Africa.

    PubMed

    Achaleke, Joseph; Brévault, Thierry

    2010-02-01

    The cotton bollworm Helicoverpa armigera (Hübner) has developed esterase-mediated resistance to pyrethroids in Central Africa. To gain a better understanding of how quickly this resistance can evolve, its genetic basis and stability were examined in a field-derived strain of H. armigera (474-fold resistant to cypermethrin). Genetic crosses between susceptible, resistant and F(1) hybrids indicated that resistance was inherited as a dominant trait (D(LD) = 0.86) and conferred by a single autosomal gene. The dominance of resistance decreased as the cypermethrin dose increased, so that resistance was incompletely dominant (D(ML) = 0.73) at the highest dose tested. Resistance (DL(50)) of the F(1) hybrid progeny significantly decreased over five generations in the absence of pyrethroid exposure. Rapid selection of resistance alleles due to dominance supports the ability of H. armigera to develop resistance to pyrethroids in Central Africa. However, associated fitness costs provide useful information for managing the evolution of resistance.

  19. RNAi silencing of the HaHMG-CoA reductase gene inhibits oviposition in the Helicoverpa armigera cotton bollworm.

    PubMed

    Wang, Zhijian; Dong, Yongcheng; Desneux, Nicolas; Niu, Changying

    2013-01-01

    RNA interference (RNAi) has considerable promise for developing novel pest control techniques, especially because of the threat of the development of resistance against current strategies. For this purpose, the key is to select pest control genes with the greatest potential for developing effective pest control treatments. The present study demonstrated that the 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase; HMGR) gene is a potential target for insect control using RNAi. HMGR is a key enzyme in the mevalonate pathway in insects. A complete cDNA encoding full length HMGR (encoding an 837-aa protein) was cloned from Helicoverpa armigera (Lepidoptera: Noctuidae). The HaHMGR (H. armigera HMGR) knockdown using systemic RNAi in vivo inhibited the fecundity of the females, effectively inhibited ovipostion, and significantly reduced vitellogenin (Vg) mRNA levels. Moreover, the oviposition rate of the female moths was reduced by 98% by silencing HaHMGR compared to the control groups. One-pair experiments showed that both the proportions of valid mating and fecundity were zero. Furthermore, the HaHMGR-silenced females failed to lay eggs (approximate 99% decrease in oviposition) in the semi-field cage performance. The present study demonstrated the potential implications for developing novel pest management strategies using HaHMGR RNAi in the control of H. armigera and other insect pests.

  20. Helicoverpa armigera baseline susceptibility to Bacillus thuringiensis Cry toxins and resistance management for Bt cotton in India.

    PubMed

    Gujar, G T; Kalia, V; Kumari, A; Singh, B P; Mittal, A; Nair, R; Mohan, M

    2007-07-01

    Transgenic cotton that produces insecticidal proteins from Bacillus thuringiensis (Bt), often referred to as Bt cotton, is widely grown in many countries. Bt cotton with a single cry1A gene and stacked also with cry2A gene has provided satisfactory protection against the damage by the lepidopteran bollworms, especially the cotton bollworm, Helicoverpa armigera (Hübner) which is considered as a key pest. The baseline susceptibility of the larvae of H. armigera to Cry1Ac and other toxins carried out in many countries has provided a basis for monitoring resistance. There is no evidence of development of field-level resistance in H. armigera leading to the failure of Bt cotton crop anywhere in the world, despite the fact that Bt cotton was grown on the largest ever area of 12.1 million hectares in 2006 and its cumulative cultivation over the last 11 years has surpassed the annual cotton area in the world. Nevertheless, the Bt resistance management has become a necessity to sustain Bt cotton and other transgenic crops in view of potential of the target insects to evolve Cry toxin resistance.

  1. Modified Artificial Diet for Rearing of Tobacco Budworm, Helicoverpa armigera, using the Taguchi Method and Derringer's Desirability Function

    PubMed Central

    Assemi, H.; Rezapanah, M.; Vafaei-Shoushtari, R.

    2012-01-01

    With the aim to improve the mass rearing feasibility of tobacco budworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), design of experimental methodology using Taguchi orthogonal array was applied. To do so, the effect of 16 ingredients of an artificial diet including bean, wheat germ powder, Nipagin, ascorbic acid, formaldehyde, oil, agar, distilled water, ascorbate, yeast, chloramphenicol, benomyl, penicillin, temperature, humidity, and container size on some biological characteristics of H. armigera was evaluated. The selected 16 factors were considered at two levels (32 experiments) in the experimental design. Among the selected factors, penicillin, container size, formaldehyde, chloramphenicol, wheat germ powder, and agar showed significant effect on the mass rearing performance. Derringer's desirability function was used for simultaneous optimization of mass rearing of tobacco budworm, H. armigera, on a modified artificial diet. Derived optimum operating conditions obtained by Derringer's desirability function and Taguchi methodology decreased larval period from 19 to 15.5 days (18.42 % improvement), decreased the pupal period from 12.29 to 11 days (10.49 % improvement), increased the longevity of adults from 14.51 to 21 days (44.72 % improvement), increased the number of eggs/female from 211.21 to 260, and increased egg hatchability from 54.2% to 72% (32.84 % improvement). The proposed method facilitated a systematic mathematical approach with a few well-defined experimental sets. PMID:23425103

  2. Dual Cry2Ab and Vip3A resistant strains of Helicoverpa armigera and Helicoverpa punctigera (Lepidoptera: Noctuidae); testing linkage between loci and monitoring of allele frequencies.

    PubMed

    Walsh, T K; Downes, S J; Gascoyne, J; James, W; Parker, T; Armstrong, J; Mahon, R J

    2014-08-01

    Considerable attention has been given to delaying the evolution of insect resistance to toxins produced by transgenic crops. The major pests of cotton in Australia are the Lepidoptera Helicoverpa armigera (Hubner, 1805) and Helicoverpa punctigera (Wallengren), and the toxins deployed in current and imminent transgenic cotton varieties are Cry1Ac, Cry2Ab and Vip3A from Bacillus thuringiensis. In this study, lines that carry alleles conferring resistance to Cry2Ab and Vip3A were isolated using F2 tests. Extensive work on the Cry2Ab resistant lines, and preliminary work on the Vip3A resistant lines, suggested a single common resistance to each toxin in both species thereby justifying the use of more efficient F1 tests as the primary means for monitoring changes over time. A potential further efficiency could be gained by developing a single resistant line that carries both types of Bt resistance. Herein we report on work with both H. armigera and H. punctigera that tests whether dual Cry2Ab-Vip3A resistant lines can be developed and, if so, whether they can be used to effectively monitor resistance frequencies. Furthermore, the creation of dual resistant lines allowed linkage between the Cry2Ab and Vip3A resistances to be investigated for H. punctigera. We show that dual resistant lines can be used to increase the efficiency of the F1 screen for recessive alleles, and that in H. punctigera there is no linkage between Cry2Ab and Vip3A resistance.

  3. Molecular Identification of Helicoverpa armigera (Lepidoptera: Noctuidae: Heliothinae) in Argentina and Development of a Novel PCR-RFLP Method for its Rapid Differentiation From H. zea and H. gelotopoeon.

    PubMed

    Arneodo, Joel D; Balbi, Emilia I; Flores, Fernando M; Sciocco-Cap, Alicia

    2015-12-01

    Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae: Heliothinae) is among the most voracious global pests of agriculture. Adults of this species were identified recently in northern Argentina by dissection of male genitalia. In this work, a rapid and simple molecular tool was designed to distinguish H. armigera from the morphologically similar indigenous bollworms Helicoverpa zea (Boddie) and Helicoverpa gelotopoeon (Dyar), regardless of the life stage. Amplification of partial COI gene with a new primer pair, and subsequent digestion with endonuclease HinfI, yielded different RFLP profiles for the three main Helicoverpa pests currently present in South America. The method was validated in Helicoverpa specimens collected across Argentina, whose identity was further corroborated by COI sequencing and phylogenetic analysis. The data reported here constitute the first molecular confirmation of this pest in the country. The survey revealed the occurrence of H. armigera in northern and central Argentina, including the main soybean- and maize-producing area.

  4. Identification of MicroRNAs in Helicoverpa armigera and Spodoptera litura Based on Deep Sequencing and Homology Analysis

    PubMed Central

    Ge, Xie; Zhang, Yong; Jiang, Jianhao; Zhong, Yi; Yang, Xiaonan; Li, Zhiqian; Huang, Yongping; Tan, Anjiang

    2013-01-01

    The current identification of microRNAs (miRNAs) in insects is largely dependent on genome sequences. However, the lack of available genome sequences inhibits the identification of miRNAs in various insect species. In this study, we used a miRNA database of the silkworm Bombyx mori as a reference to identify miRNAs in Helicoverpa armigera and Spodoptera litura using deep sequencing and homology analysis. Because all three species belong to the Lepidoptera, the experiment produced reliable results. Our study identified 97 and 91 conserved miRNAs in H. armigera and S. litura, respectively. Using the genome of B. mori and BAC sequences of H. armigera as references, 1 novel miRNA and 8 novel miRNA candidates were identified in H. armigera, and 4 novel miRNA candidates were identified in S. litura. An evolutionary analysis revealed that most of the identified miRNAs were insect-specific, and more than 20 miRNAs were Lepidoptera-specific. The investigation of the expression patterns of miR-2a, miR-34, miR-2796-3p and miR-11 revealed their potential roles in insect development. miRNA target prediction revealed that conserved miRNA target sites exist in various genes in the 3 species. Conserved miRNA target sites for the Hsp90 gene among the 3 species were validated in the mammalian 293T cell line using a dual-luciferase reporter assay. Our study provides a new approach with which to identify miRNAs in insects lacking genome information and contributes to the functional analysis of insect miRNAs. PMID:23289012

  5. Relative Fitness of Helicoverpa armigera (Lepidoptera: Noctuidae) on Seven Host Plants: A Perspective for IPM in Brazil

    PubMed Central

    Reigada, C.; Guimarães, K. F.; Parra, J. R. P.

    2016-01-01

    The cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a widespread pest of many cultivated and wild plants in Europe, Africa, Asia, and Australia. In 2013, this species was reported in Brazil, attacking various host crops in the midwestern and northeastern regions of the country and is now found countrywide. Aiming to understand the effects of different host plants on the life cycle of H. armigera, we selected seven species of host plants that mature in different seasons and are commonly grown in these regions: cotton (Gossypium hirsutum, “FM993”), corn (Zea mays, “2B587”), soybean (Glycine max, “99R01”), rattlepods (Crotalaria spectabilis), millet (Pennisetum glaucum, “ADR300”), sorghum (Sorghum bicolor, “AGROMEN70G35”), and cowpea (Vigna unguiculata, “SEMPRE VERDE”). The development time of immatures, body weight, survivorship, and fecundity of H. armigera were evaluated on each host plant under laboratory conditions. The bollworms did not survive on corn, millet, or sorghum and showed very low survival rates on rattlepods. Survival rates were highest on soybean, followed by cotton and cowpea. The values for relative fitness found on soybean, cotton, cowpea, and rattlepods were 1, 0.5, 0.43, and 0.03, respectively. Survivorship, faster development time, and fecundity on soybean, cotton, and cowpea were positively correlated. Larger pupae and greater fecundity were found on soybean and cotton. The results indicated that soybean, cotton, and cowpea are the most suitable plants to support the reproduction of H. armigera in the field. PMID:26798139

  6. Comparative feeding performance and digestive physiology of Helicoverpa armigera (Lepidoptera: Noctuidae) larvae-fed 11 corn hybrids.

    PubMed

    Hosseininejad, A S; Naseri, B; Razmjou, J

    2015-01-01

    This study aimed to evaluate the feeding responses and digestive proteolytic and amylolytic activity of Helicoverpa armigera (Hübner) on 11 corn (Zea mays L.) hybrids at 25 ± 1°C, 65 ± 5% relative humidity (RH), and a photoperiod of 16:8 (L:D) h. The fourth- and fifth-instar larvae fed on hybrid K47*K19 had the highest weight of food consumption and those reared on hybrid KSC705 had the lowest value of food consumption. The highest weight gain of the larvae was observed when H. armigera were fed hybrid KLM78*MO17 and lowest when they were fed hybrids K36 * MO17, KSC705, and K35 * K36. Pupal weight of H. armigera was heaviest when larvae were fed hybrid K47*K19 and lightest when they were fed hybrid KSC705. The highest proteolytic activity of the fourth-instar larvae was observed when they were fed hybrid KSC705, and the lowest activity was observed when they were fed hybrid K47*A67. Fifth-instar larvae that fed on hybrid K47*K19 showed the highest proteolytic activity. Fourth-instar larvae that fed on hybrid K36*MO17 showed the highest amylase activity. The fifth-instar larvae fed on hybrid K47*A67 showed the maximum amylase activity and those reared on the K48*K18 showed the minimum activity. Our results indicated that K36 * MO17, KSC705, and K48 * K18 were the most unsuitable hybrids for feeding H. armigera. © The Author 2015. Published by Oxford University Press on behalf of the Entomological Society of America.

  7. Comparative Feeding Performance and Digestive Physiology of Helicoverpa armigera (Lepidoptera: Noctuidae) Larvae-Fed 11 Corn Hybrids

    PubMed Central

    Hosseininejad, A. S.; Naseri, B.; Razmjou, J.

    2015-01-01

    This study aimed to evaluate the feeding responses and digestive proteolytic and amylolytic activity of Helicoverpa armigera (Hübner) on 11 corn (Zea mays L.) hybrids at 25 ± 1°C, 65 ± 5% relative humidity (RH), and a photoperiod of 16:8 (L:D) h. The fourth- and fifth-instar larvae fed on hybrid K47*K19 had the highest weight of food consumption and those reared on hybrid KSC705 had the lowest value of food consumption. The highest weight gain of the larvae was observed when H. armigera were fed hybrid KLM78*MO17 and lowest when they were fed hybrids K36 * MO17, KSC705, and K35 * K36. Pupal weight of H. armigera was heaviest when larvae were fed hybrid K47*K19 and lightest when they were fed hybrid KSC705. The highest proteolytic activity of the fourth-instar larvae was observed when they were fed hybrid KSC705, and the lowest activity was observed when they were fed hybrid K47*A67. Fifth-instar larvae that fed on hybrid K47*K19 showed the highest proteolytic activity. Fourth-instar larvae that fed on hybrid K36*MO17 showed the highest amylase activity. The fifth-instar larvae fed on hybrid K47*A67 showed the maximum amylase activity and those reared on the K48*K18 showed the minimum activity. Our results indicated that K36 * MO17, KSC705, and K48 * K18 were the most unsuitable hybrids for feeding H. armigera. PMID:25688090

  8. Isolation and Characterization of Gut Bacterial Proteases Involved in Inducing Pathogenicity of Bacillus thuringiensis Toxin in Cotton Bollworm, Helicoverpa armigera

    PubMed Central

    Regode, Visweshwar; Kuruba, Sreeramulu; Mohammad, Akbar S.; Sharma, Hari C.

    2016-01-01

    Bacillus thuringiensis toxin proteins are deployed in transgenic plants for pest management. The present studies were aimed at characterization of gut bacterial proteases involved in activation of inactive Cry1Ac protoxin (pro-Cry1Ac) to active toxin in Helicoverpa armigera. Bacterial strains were isolated from H. armigera midgut and screened for their proteolytic activation toward pro-Cry1Ac. Among 12 gut bacterial isolates seven isolates showed proteolytic activity, and proteases from three isolates (IVS1, IVS2, and IVS3) were found to be involved in the proteolytic conversion of pro-Cry1Ac into active toxin. The proteases from IVS1, IVS2, and IVS3 isolates were purified to 11.90-, 15.50-, and 17.20-fold, respectively. The optimum pH and temperature for gut bacterial protease activity was 8.0 and 40°C. Maximum inhibition of total proteolytic activity was exerted by phenylmethane sulfonyl fluoride followed by EDTA. Fluorescence zymography revealed that proteases from IVS1, IVS2, and IVS3 were chymotrypsin-like and showing protease band at ~15, 65, and 15 kDa, respectively. Active Cry1Ac formed from processing pro-Cry1Ac by gut bacterial proteases exhibited toxicity toward H. armigera. The gut bacterial isolates IVS1, IVS2, and IVS3 showed homology with B. thuringiensis (CP003763.1), Vibrio fischeri (CP000020.2), and Escherichia coli (CP011342.1), respectively. Proteases produced by midgut bacteria are involved in proteolytic processing of B. thuringiensis protoxin and play a major role in inducing pathogenicity of B. thuringiensis toxins in H. armigera. PMID:27766093

  9. Cloning and characterisation of NADPH-dependent cytochrome P450 reductase gene in the cotton bollworm, Helicoverpa armigera.

    PubMed

    Zhao, Chunqing; Tang, Tao; Feng, Xiaoyun; Qiu, Lihong

    2014-01-01

    Previous studies in our laboratory showed that cytochrome P450 CYP6B7 plays a critical role in a Handan fenvalerate resistant strain (HDFR) of Helicoverpa armigera. As an important component of P450 enzyme systems, cytochrome P450 reductase (CPR) plays an essential role in transferring electrons from NADPH to the P450-substrate complex. However, little information about CPR in H. armigera (HaCPR) has been reported. A full-length cDNA (3525 bp) of HaCPR was cloned. The open reading frame of the HaCPR gene encoded 687 amino acids and shared 27.87-95.21% identities with other known CPRs. Bioinformatic analysis showed that HaCPR is a transmembrane protein with Mw of 77.4 kDa and contains conserved features. The results of real-time quantitative polymerase chain reaction showed that the expression level of HaCPR mRNA was 1.84-fold higher in midgut of 5th instars of the Handan susceptible strain than that in pupae, and the level in the midgut of HDFR strain was 2.02-fold higher than that of the Handan susceptible strain. The levels of HaCPR mRNA were induced by phenobarbital at concentrations of 2 and 4 mg g(-1) , which enhanced 5.20- and 17.45-fold, respectively, compared to that of the control after 48 h of phenobarbital treatment. The results indicate that HaCPR is important for the development of H. armigera and may play an essential role in the P450-mediated insecticide resistance of H. armigera to fenvalerate. © 2013 Society of Chemical Industry.

  10. The Inheritance of the Pheromone Sensory System in Two Helicoverpa Species: Dominance of H. armigera and Possible Introgression from H. assulta

    PubMed Central

    Xu, Meng; Dong, Jun-Feng; Wu, Han; Zhao, Xin-Cheng; Huang, Ling-Qiao; Wang, Chen-Zhu

    2017-01-01

    Hybridization of sympatric closely related species may sometimes lead to introgression and speciation. The sister species Helicoverpa armigera and Helicoverpa assulta both use (Z)-11-hexadecenal and (Z)-9-hexadecenal as sex pheromone components but in reversed ratios. Female H. armigera and male H. assulta could hybridize and produce fertile male hybrids, which can then backcross with females of the two parent species to get backcross lines in the laboratory. In this study, we compared the olfactory responses to pheromone compounds in the periphery and in the antennal lobes (ALs) of males of the two species, as well as of their hybrids and backcrosses. Single-sensillum recordings were carried out to explore characteristics of male-specific sensilla on the antennae, and in vivo calcium imaging combined with digital 3D-reconstruction was used to describe what happens in the macroglomerular complex (MGC) of the AL. The results show that the population ratio of the two male-specific types of olfactory sensory neurons responding to two sex pheromone components are controlled by a major gene, and that the allele of H. armigera is dominant. Consistently, the study of the representative areas activated by sex pheromone components in the ALs further support the dominance of H. armigera. However, the topological structure of the MGC in the hybrid was similar but not identical to that in H. armigera. All subtypes of male-specific sensilla identified in the two species were found in the male hybrids and backcrosses. Moreover, two new subtypes with broader response spectra (the expanded A subtype and the expanded C subtype) emerged in the hybrids. Based on the inheritance pattern of the pheromone sensory system, we predict that when hybridization of female H. armigera and male H. assulta occurs in the field, male hybrids would readily backcross with female H. armigera, and introgression might occur from H. assulta into H. armigera through repeated backcrossing. PMID:28119570

  11. Gene cloning and expression of cadherin in midgut of Helicoverpa armigera and its Cry1A binding region.

    PubMed

    Wang, Guirong; Wu, Kongming; Liang, Gemei; Guo, Yuyuan

    2005-08-01

    Cadherins belong to one of the families of animal glycoproteins responsible for calcium-dependent cell-cell adhesion. Recent literatures showed that the cadherin-like in midgut of several insects served as the receptor of Bt toxin Cry1A and the variation of cadherin-like is related to insect's resistance to Cry1A. The full-length cDNA encoding cadherin-like of Helicoverpa armigera is cloned by degenerate PCR and RACE techniques and the gene was designated as BtR-harm, which is 5581 bp in full-length, encoding 1730 amino acid residues (BtR-harm was deposited in GenBank and the accession number is AF519180). Its predicted molecular weight and isoelectric point were 195.39 kDa and 4.23, respectively. The inferred amino acid sequence includes a signal sequence, 11 cadherin repeats, a membrane-proximal region, a transmembrane region and a cytoplasmic region. Sequence analysis indicated that the deduced protein sequence was most similar to the cadherin-like from Heliothis virescens with 84.2% identity and highly similar to three other lepidopteran cadherin from Bombyx mori, Manduca sexta and Pectinophora gossypiella, with the sequence identities of 60.3.6%, 57.5% and 51.0%, respectively. The cDNA encoding cadherin gene was expressed successfully in E. coli and the recombinant proteins can bind with Cry1Ac. Truncation analysis and binding experiment of BtR-harm revealed that the Cry1A binding region was a contiguous 244-amino acid sequence, which located between amino acid 1217 and 1461. Semi-quantitative RT-PCR analysis showed that BtR-harm was highly expressed in midgut of H. armigera, very low expressed in foregut and hindgut and was not expressed in other tissues. After H. armigera producing resistance to Cry1Ac, the expression quantity of BtR-harm significantly decreased in midgut of H. armigera. It is the first confirmation that BtR-harm can function as receptor of Cry1Ac in H. armigera and the binding region was located on a contiguous 244 amino acid sequence

  12. Bacillus thuringiensis insecticidal crystal proteins affect lifespan and reproductive performance of Helicoverpa armigera and Spodoptera exigua adults.

    PubMed

    Zhang, Ying; Ma, Yan; Wan, Pin-Jun; Mu, Li-Li; Li, Guo-Qing

    2013-04-01

    Being delivered as sprays or expressed in plant, Bacillus thuringiensis (Bt) crystalline proteins (Cry toxins) display insecticidal activities against numerous Lepidopteran, Dipteran, and Coleopteran larvae. Comparative study of toxicities of Bt Cry toxins between larvae and adults may afford important new insights into the interactions of the toxins with receptor proteins in host insect, and represent intriguing targets for the control of insect pests. However, the effectiveness of Bt Cry toxins in insect adults has paid less attention. In the present article, the effectiveness of Cry1Ac and Cry1Ca on lifespans and reproductive performance of Helicoverpa armigera (Hübner) and Spodoptera exigua (Hübner) adults were evaluated by in vivo experiments. Considering transgenic plants express modified, truncated versions of cry genes yielding active toxin fragment, we used activated Bt toxins at the concentration of 500, 100, and 20 microg/ml in a 10% sucrose aquous solution. At the highest concentration, Cry1Ac and Cry1Ca shortened 48.1 and 48.9% of H. armigera female lifespan, and 43.5 and 38.5% of S. exigua female lifespan, and they reduced 37.8 and 40.3%, and 50.5 and 47.4% of H. armigera and S. exigua male lifespans respectively. Bt toxins negatively affected copulation. Exposure to 500 microg/ml of Cry1Ac and Cry1Ca greatly reduced 50.0 and 46.8%, and 58.7 and 57.3% spermatophore acceptance by H. armigera and S. exigua females, respectively. Similarly, Cry1Ac and Cry1Ca exposure decreased 40.0 and 50.3%, and 61.3 and 60.0% of spermatophore transfer by H. armigera and S. exigua males, respectively. Moreover, exposure females rather than males to 500 microg/ml of Cry1Ac and Cry1Ca significantly dropped 57.5 and 57.5% of the number of eggs laid by H. armigera, and 35.4 and 45.8% of the number of egg masses deposited by S. exigua. In contrast, both Cry1Ac and Cry1Ca did not negatively influence the egg hatchability. At the middle and the lowest concentrations, however

  13. Effects of elevated CO2 on the fitness and potential population damage of Helicoverpa armigera based on two-sex life table.

    PubMed

    Liu, Jinping; Huang, Wenkun; Chi, Hsin; Wang, Chonghui; Hua, Hongxia; Wu, Gang

    2017-04-25

    We evaluated the direct effects of three different atmospheric CO2 concentrations (380 ppm, 550 ppm and 750 ppm) on the population parameters of the cotton bollworm, Helicoverpa armigera fed an artificial diet. The life history and fitness of H. armigera were analyzed using an age-stage, two-sex life table. Our results showed significantly longer larval durations and lower female pupal weight under elevated CO2 than under ambient CO2. Additionally, the fecundity of H. armigera was lower under elevated CO2 than under ambient CO2. H. armigera reared under elevated CO2 conditions showed lower intrinsic and finite rates of increase but higher net consumption and finite consumption rates than H. armigera reared under ambient CO2 conditions. According to population projections, a much smaller total population size and reduced consumption capacities would be expected in an elevated CO2 atmosphere due to higher mortality and lower growth rate compared with ambient CO2 levels. These results indicate that the fitness of and potential damage caused by H. armigera would be affected by increased CO2 relative to ambient CO2 concentrations. Additional studies on the long-term direct and indirect effects of elevated CO2 levels on H. armigera are still needed.

  14. Species From the Heliothinae Complex (Lepidoptera: Noctuidae) in Tucumán, Argentina, an Update of Geographical Distribution of Helicoverpa armigera

    PubMed Central

    Murúa, M. Gabriela; Cazado, Lucas E.; Casmuz, Augusto; Herrero, M. Inés; Villagrán, M. Elvira; Vera, Alejandro; Sosa-Gómez, Daniel R.; Gastaminza, Gerardo

    2016-01-01

    The Heliothinae complex in Argentina encompasses Helicoverpa gelotopoeon (Dyar), Helicoverpa zea (Boddie), Helicoverpa armigera (Hübner), and Chloridea virescens (Fabricius). In Tucumán, the native species H. gelotopoeon is one of the most voracious soybean pests and also affects cotton and chickpea, even more in soybean-chickpea succession cropping systems. Differentiation of the Heliothinae complex in the egg, larva, and pupa stages is difficult. Therefore, the observation of the adult wing pattern design and male genitalia is useful to differentiate species. The objective of this study was to identify the species of the Heliothinae complex, determine population fluctuations of the Heliothinae complex in soybean and chickpea crops using male moths collected in pheromone traps in Tucuman province, and update the geographical distribution of H. armigera in Argentina. The species found were H. gelotopoeon, H. armigera, H. zea, and C. virescens. Regardless of province, county, crop, and year, the predominant species was H. gelotopoeon. Considering the population dynamics of H. gelotopoeon and H. armigera in chickpea and soybean crops, H. gelotopoeon was the most abundant species in both crops, in all years sampled, and the differences registered were significant. On the other hand, according to the Sistema Nacional Argentino de Vigilancia y Monitoreo de Plagas (SINAVIMO) database and our collections, H. armigera was recorded in eight provinces and 20 counties of Argentina, and its larvae were found on soybean, chickpea, sunflower crops and spiny plumeless thistle (Carduus acanthoides). This is the first report of H. armigera in sunflower and spiny plumeless thistle in Argentina. PMID:27324588

  15. Species From the Heliothinae Complex (Lepidoptera: Noctuidae) in Tucumán, Argentina, an Update of Geographical Distribution of Helicoverpa armigera.

    PubMed

    Murúa, M Gabriela; Cazado, Lucas E; Casmuz, Augusto; Herrero, M Inés; Villagrán, M Elvira; Vera, Alejandro; Sosa-Gómez, Daniel R; Gastaminza, Gerardo

    2016-01-01

    The Heliothinae complex in Argentina encompasses Helicoverpa gelotopoeon (Dyar), Helicoverpa zea (Boddie), Helicoverpa armigera (Hübner), and Chloridea virescens (Fabricius). In Tucumán, the native species H. gelotopoeon is one of the most voracious soybean pests and also affects cotton and chickpea, even more in soybean-chickpea succession cropping systems. Differentiation of the Heliothinae complex in the egg, larva, and pupa stages is difficult. Therefore, the observation of the adult wing pattern design and male genitalia is useful to differentiate species. The objective of this study was to identify the species of the Heliothinae complex, determine population fluctuations of the Heliothinae complex in soybean and chickpea crops using male moths collected in pheromone traps in Tucuman province, and update the geographical distribution of H. armigera in Argentina. The species found were H. gelotopoeon, H. armigera, H. zea, and C. virescens. Regardless of province, county, crop, and year, the predominant species was H. gelotopoeon Considering the population dynamics of H. gelotopoeon and H. armigera in chickpea and soybean crops, H. gelotopoeon was the most abundant species in both crops, in all years sampled, and the differences registered were significant. On the other hand, according to the Sistema Nacional Argentino de Vigilancia y Monitoreo de Plagas (SINAVIMO) database and our collections, H. armigera was recorded in eight provinces and 20 counties of Argentina, and its larvae were found on soybean, chickpea, sunflower crops and spiny plumeless thistle (Carduus acanthoides). This is the first report of H. armigera in sunflower and spiny plumeless thistle in Argentina.

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

    PubMed Central

    Hu, Rongfeng; Wang, Jiehua; Ji, Yan; Song, Yingjin; Yang, Shaohui

    2012-01-01

    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. PMID:23226090

  17. Effects of climate change on overwintering pupae of the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

    NASA Astrophysics Data System (ADS)

    Huang, Jian; Li, Jing

    2015-07-01

    Climate change significantly affects insects' behaviors. Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is one of the most serious insect pests in the world. Much is known about the survival of the overwintering population and spring emergence of H. armigera. However, little is known about the effects of climate change on overwintering and spring emergence of H. armigera. This study investigated the effects of changes of air and soil temperatures and precipitation on overwintering pupae of H. armigera by analyzing historical data from Magaiti County in northwest China using statistical methods. The results showed that during the period of 1989-2006, the climate warming advanced the first-appearance date of overwintering pupae eclosion (FD) and end date of overwintering pupae eclosion (ED) by 1.276 and 0.193 days per year, respectively; the duration between the FD and ED (DFEPE) was prolonged by 1.09 days per year, which resulted in more eclosion of overwintering pupae. For a 1 °C increase in the maximum air temperature ( T max) in winter, the FD became earlier by 3.234 days. Precipitation in winter delayed the FD and ED and produced little relative influence on DFEPE. A 1-mm increase of precipitation in winter delayed the FD and ED by 0.850 and 0.494 days, respectively. Mean air temperature ( T mean) in March, with a 41.3 % relative influence, precipitation in winter, with a 49.0 % relative influence, and T mean in March, with a 37.5 % relative influence, were the major affecting factors on FD, ED, and DFEPE, respectively. T max in February with a 53.0 % relative influence was the major affecting factor on the mortality of overwintering pupae (MOP). Increased soil temperatures in October and November and autumn and air temperatures in winter could decrease the MOP, though the relative influences were lower than T max in February. Increased precipitation in winter increased the MOP, but the relative influence was only 4.2 % because of little precipitation

  18. Integrated proteomic and metabolomic analysis of larval brain associated with diapause induction and preparation in the cotton bollworm, Helicoverpa armigera.

    PubMed

    Zhang, Qi; Lu, Yu-Xuan; Xu, Wei-Hua

    2012-02-03

    Diapause is a developmental arrest that allows an organism to survive unfavorable environmental conditions and is induced by environmental signals at a certain sensitive developmental stage. In Helicoverpa armigera, the larval brain receives the environmental signals for diapause induction and then regulates diapause entry at the pupal stage. Here, combined proteomic and metabolomic differential display analysis was performed on the H. armigera larval brain. Using two-dimensional electrophoresis, it was found that 22 proteins were increased and 27 proteins were decreased in the diapause-destined larval brain, 37 of which were successfully identified by MALDI-TOF/TOF mass spectrometry. RT-PCR and Western blot analyses showed that the expression levels of the differentially expressed proteins were consistent with the 2-DE results. Furthermore, a total of 49 metabolites were identified in the larval brain by GC-MS analysis, including 4 metabolites at high concentrations and 14 metabolites at low concentrations. The results gave us a clue to understand the governing molecular events of the prediapause phase. Those differences that exist in the induction phase of diapause-destined individuals are probably relevant to a special memory mechanism for photoperiodic information storage, and those differences that exist in the preparation phase are likely to regulate accumulation of specific energy reserves in diapause-destined individuals.

  19. Transgenic Cotton Plants Expressing the HaHR3 Gene Conferred Enhanced Resistance to Helicoverpa armigera and Improved Cotton Yield.

    PubMed

    Han, Qiang; Wang, Zhenzhen; He, Yunxin; Xiong, Yehui; Lv, Shun; Li, Shupeng; Zhang, Zhigang; Qiu, Dewen; Zeng, Hongmei

    2017-08-30

    RNA interference (RNAi) has been developed as an efficient technology. RNAi insect-resistant transgenic plants expressing double-stranded RNA (dsRNA) that is ingested into insects to silence target genes can affect the viability of these pests or even lead to their death. HaHR3, a molt-regulating transcription factor gene, was previously selected as a target expressed in bacteria and tobacco plants to control Helicoverpa armigera by RNAi technology. In this work, we selected the dsRNA-HaHR3 fragment to silence HaHR3 in cotton bollworm for plant mediated-RNAi research. A total of 19 transgenic cotton lines expressing HaHR3 were successfully cultivated, and seven generated lines were used to perform feeding bioassays. Transgenic cotton plants expressing dsHaHR3 were shown to induce high larval mortality and deformities of pupation and adult eclosion when used to feed the newly hatched larvae, and 3rd and 5th instar larvae of H. armigera. Moreover, HaHR3 transgenic cotton also demonstrated an improved cotton yield when compared with controls.

  20. Electrophysiological and behavioral responses of female Helicoverpa armigera to compounds identified in flowers of African marigold, Tagetes erecta.

    PubMed

    Bruce, T J; Cork, A

    2001-06-01

    Seven electrophysiologically active compounds were detected in air-entrained headspace samples of live flowers of Tagetes erecta analyzed by gas chromatography (GC) linked to a female Helicoverpa armigera electroantennograph (EAG) using polar and nonpolar capillary columns. These compounds were subsequently identified using GC linked to mass spectrometry as benzaldehyde, (S)-(-)-limonene, (R,S)-(+/-)-linalool, (E)-myroxide, (Z)-beta-ocimene, phenylacetaldehyde, and (R)-(-)-piperitone. Electrophysiological activity was confirmed by EAG with a 1-microg dose of each compound on filter paper eliciting EAG responses that were significantly greater than the solvent control response from female moths. Wind-tunnel bioassays with T. erecta headspace samples, equivalent to 0.4 flower/hr emission from a live flower, elicited a significant increase in the number of upwind approaches from female H. armigera relative to a solvent control. Similarly, a seven-component synthetic blend of EAG-active compounds identified from T. erecta presented in the same ratio (1.0:1.6:0.7:1.4:0.4:5.0:2.7, respectively) and concentration (7.2 microg) as found in the natural sample elicited a significant increase in the number of upwind approaches relative to a solvent control during a 12-min bioassay that was equivalent to that elicited by the natural T. erecta floral volatiles.

  1. Effect of Larvae Treated with Mixed Biopesticide Bacillus thuringiensis - Abamectin on Sex Pheromone Communication System in Cotton Bollworm, Helicoverpa armigera

    PubMed Central

    Shen, Li-Ze; Chen, Peng-Zhou; Xu, Zhi-Hong; Deng, Jian-Yu; Harris, Marvin-K; Wanna, Ruchuon; Wang, Fu-Min; Zhou, Guo-Xin; Yao, Zhang-Liang

    2013-01-01

    Third instar larvae of the cotton bollworm (Helicoverpa armigera) were reared with artificial diet containing a Bacillus thuringiensis - abamectin (BtA) biopesticide mixture that resulted in 20% mortality (LD20). The adult male survivors from larvae treated with BtA exhibited a higher percentage of “orientation” than control males but lower percentages of “approaching” and “landing” in wind tunnel bioassays. Adult female survivors from larvae treated with BtA produced higher sex pheromone titers and displayed a lower calling percentage than control females. The ratio of Z-11-hexadecenal (Z11–16:Ald) and Z-9-hexadecenal (Z9–16:Ald) in BtA-treated females changed and coefficients of variation (CV) of Z11–16:Ald and Z9–16:Ald were expanded compared to control females. The peak circadian calling time of BtA-treated females occurred later than that of control females. In mating choice experiment, both control males and BtA-treated males preferred to mate with control females and a portion of the Bt-A treated males did not mate whereas all control males did. Our Data support that treatment of larvae with BtA had an effect on the sex pheromone communication system in surviving H.armigera moths that may contribute to assortative mating. PMID:23874751

  2. Role of induced glutathione-S-transferase from Helicoverpa armigera (Lepidoptera: Noctuidae) HaGST-8 in detoxification of pesticides.

    PubMed

    Labade, Chaitali P; Jadhav, Abhilash R; Ahire, Mehul; Zinjarde, Smita S; Tamhane, Vaijayanti A

    2017-09-15

    The present study deals with glutathione-S-transferase (GST) based detoxification of pesticides in Helicoverpa armigera and its potential application in eliminating pesticides from the environment. Dietary exposure of a pesticide mixture (organophosphates - chlorpyrifos and dichlorvos, pyrethroid - cypermethrin; 2-15ppm each) to H. armigera larvae resulted in a dose dependant up-regulation of GST activity and gene expression. A variant GST from H. armigera (HaGST-8) was isolated from larvae fed with 10ppm pesticide mixture and it was recombinantly expressed in yeast (Pichia pastoris HaGST-8). HaGST-8 had a molecular mass of 29kDa and was most active at pH 9 at 30°C. GC-MS and LC-HRMS analysis validated that HaGST-8 was effective in eliminating organophosphate type of pesticides and partially reduced the cypermethrin content (53%) from aqueous solutions. Unlike the untransformed yeast, P. pastoris HaGST-8 grew efficiently in media supplemented with pesticide mixtures (200 and 400ppm each pesticide) signifying the detoxification ability of HaGST-8. The amino acid sequence of HaGST-8 and the already reported sequence of HaGST-7 had just 2 mismatches. The studies on molecular interaction strengths revealed that HaGST-8 had stronger binding affinities with organophosphate, pyrethroid, organochloride, carbamate and neonicotinoid type of pesticides. The abilities of recombinant HaGST-8 to eliminate pesticides and P. pastoris HaGST-8 to grow profusely in the presence of high level of pesticide content can be applied for removal of such residues from food, water resources and bioremediation. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Molecular and insecticidal characterization of Vip3A protein producing Bacillus thuringiensis strains toxic against Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Lone, Showkat Ahmad; Yadav, Radha; Malik, Abdul; Padaria, Jasdeep Chatrath

    2016-02-01

    Vegetative insecticidal proteins (Vip) represent the second generation of insecticidal proteins produced by Bacillus thuringiensis (Bt) during the vegetative growth stage of growth. Bt-based biopesticides are recognized as viable alternatives to chemical insecticides; the latter cause environmental pollution and lead to the emergence of pest resistance. To perform a systematic study of vip genes encoding toxic proteins, a total of 30 soil samples were collected from diverse locations of Kashmir valley, India, and characterized by molecular and analytical methods. Eighty-six colonies showing Bacillus-like morphology were selected. Scanning electron microscopy observations confirmed the presence of different crystal shapes, and PCR analysis of insecticidal genes revealed a predominance of the lepidopteran-specific vip3 (43.18%) gene followed by coleopteran-specific vip1 (22.72%) and vip2 (15.90%) genes in the isolates tested. Multi-alignment of the deduced amino acid sequences revealed that vip3 sequences were highly conserved, whereas vip1 and vip2 showed adequate differences in amino acid sequences compared with already reported sequences. Screening for toxicity against Helicoverpa armigera larvae was performed using partially purified soluble fractions containing Vip3A protein. The mortality levels observed ranged between 70% and 96.6% in the isolates. The LC50 values of 2 of the native isolates, JK37 and JK88, against H. armigera were found to be on par with that of Bt subsp. kurstaki HD1, suggesting that these isolates could be developed as effective biopesticides against H. armigera.

  4. Multiple Resistances Against Formulated Organophosphates, Pyrethroids, and Newer-Chemistry Insecticides in Populations of Helicoverpa armigera (Lepidoptera: Noctuidae) from Pakistan.

    PubMed

    Qayyum, Mirza Abdul; Wakil, Waqas; Arif, Muhammad Jalal; Sahi, Shahbaz Talib; Saeed, Noor Abid; Russell, Derek Allan

    2015-02-01

    Field populations of Helicoverpa armigera Hübner from 15 localities across the Punjab, Pakistan, were assessed by the leaf dip method for resistance against formulated organophosphates, pyrethroids, and newer insecticide groups. Resistance levels in H. armigera have been incrementally increasing for organophosphate and pyrethroid insecticides after decades of use in Pakistan. Resistance ratios (RRs) documented for organophosphates were 24- to 116-fold for profenofos and 22- to 87-fold for chlorpyrifos. For pyrethroids, RRs were 3- to 69-fold for cypermethrin and 3- to 27-fold for deltamethrin. Resistance levels against newer chemistries were 2- to 24-fold for chlorfenapyr, 1- to 22-fold for spinosad, 1- to 20-fold for indoxacarb, 1- to 18-fold for abamectin, and 1- to 16-fold for emamectin benzoate. Resistant populations of H. armigera were mainly in the southern part of the Punjab, Pakistan. The most resistant populations were collected from Pakpattan, Multan, and Muzzafargarh. Of the nine insecticides tested, LC50 and LC90 values were lower for newer insecticide groups; resistance levels were moderate to very high against organophosphates, very low to high against pyrethroids, and very low to low against the newer-chemistry insecticides. These findings suggest that the newer-chemistry insecticides with different modes of action could be included in insecticide rotations or replace the older insecticides. Supplementing the use of synthetic insecticides with safer alternatives could help to successfully lower the farmer's reliance on insecticides and the incidence of resistance due to repeated use of insecticides against major insect pests. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  5. Identification and expression analysis of NADH-cytochrome b₅ reductase gene in the cotton bollworm, Helicoverpa armigera.

    PubMed

    Zhao, Chunqing; Tang, Tao; Liu, Jiqin; Feng, Xiaoyun; Qiu, Lihong

    2012-12-10

    NADH-cytochrome b(5) reductase (CBR) is one of the most important components of cytochrome P450s, which play an essential role in the detoxification of xenobiotics as well as insecticide resistance in insect pest. In the present study, two novel full-length cDNAs of CBR of the cotton bollworm, Helicoverpa armigera (Hübner) were amplified by means of reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. The sequencing results showed that the transcripts were 1809bp and 1518bp for HaCBR1 and HaCBR2, respectively, including 969bp and 939bp of complete open reading frame (ORF), which encoded 322 and 312 amino acids respectively. The putative structure and function of HaCBR1 and HaCBR2 were preliminarily analyzed by SMART program. HaCBR1 and HaCBR2 (GenBank accession numbers: HQ638220 and HQ190046HQ638220HQ190046) showed high identities with CBRs of other species. The expression of HaCBR1 and HaCBR2 mRNA was detected by real-time quantitative polymerase chain reaction (RT-qPCR) in most developmental stages of H. armigera with the exception of eggs, as well as in tissues such as cuticle, fatbody and midgut. The expression level of the two genes was significantly induced by phenobarbital (PB). These results would contribute to the understanding of CBR function in H. armigera and provide information for further study on the interactions of different components of cytochrome P450 enzyme systems. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Quercetin interacts with Cry1Ac protein to affect larval growth and survival of Helicoverpa armigera.

    PubMed

    Li, Zhen; Guan, Xiumin; Michaud, J P; Zhang, Qingwen; Liu, Xiaoxia

    2016-07-01

    Bt cotton has been widely planted in China for over a decade to control H. armigera, but field surveys indicate increasing resistance in the pest. It has been speculated that accumulating plant secondary compounds in mature cotton may interact with Bt toxins and affect the toxicity of Bt to H. armigera. Both quercetin, one of the main flavonoids in cotton, and the Bt toxin Cry1Ac protein had significant negative effects on the growth, development and survival of H. armigera when added singly to artificial diet, but their effects were inhibited when added in combination. Quercetin was antagonistic to Cry1Ac toxicity at all tested concentrations. The accumulation of quercetin might be one factor contributing to the reduced toxicity of mature Bt cotton plants to H. armigera, and could partially explain the reduced efficacy of Cry1Ac in controlling this pest in the field. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  7. High Susceptibility to Cry1Ac and Low Resistance Allele Frequency Reduce the Risk of Resistance of Helicoverpa armigera to Bt Soybean in Brazil

    PubMed Central

    Bacalhau, Fabiana B.; Amado, Douglas; Carvalho, Renato A.; Martinelli, Samuel; Head, Graham P.; Omoto, Celso

    2016-01-01

    The Old World bollworm, Helicoverpa armigera (Hübner), was recently introduced into Brazil, where it has caused extensive damage to cotton and soybean crops. MON 87701 × MON 89788 soybean, which expresses the Bt protein Cry1Ac, was recently deployed in Brazil, providing high levels of control against H. armigera. To assess the risk of resistance to the Cry1Ac protein expressed by MON 87701 × MON 89788 soybean in Brazil, we conducted studies to evaluate the baseline susceptibility of H. armigera to Cry1Ac, in planta efficacy including the assessment of the high-dose criterion, and the initial resistance allele frequency based on an F2 screen. The mean Cry1Ac lethal concentration (LC50) ranged from 0.11 to 1.82 μg·mL−1 of diet among all H. armigera field populations collected from crop seasons 2013/14 to 2014/15, which indicated about 16.5-fold variation. MON 87701 × MON 89788 soybean exhibited a high level of efficacy against H. armigera and most likely met the high dose criterion against this target species in leaf tissue dilution bioassays up to 50 times. A total of 212 F2 family lines of H. armigera were established from field collections sampled from seven locations across Brazil and were screened for the presence of MON 87701 × MON 89788 soybean resistance alleles. None of the 212 families survived on MON 87701 × MON 89788 soybean leaf tissue (estimated allele frequency = 0.0011). The responses of H. armigera to Cry1Ac protein, high susceptibility to MON 87701 × MON 89788 soybean, and low frequency of resistance alleles across the main soybean-producing regions support the assumptions of a high-dose/refuge strategy. However, maintenance of reasonable compliance with the refuge recommendation will be essential to delay the evolution of resistance in H. armigera to MON 87701 × MON 89788 soybean in Brazil. PMID:27532632

  8. Broad-scale suppression of cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae), associated with Bt cotton crops in Northern New South Wales, Australia.

    PubMed

    Baker, G H; Tann, C R

    2017-04-01

    The cotton bollworm, Helicoverpa armigera, is a major pest of many agricultural crops in several countries, including Australia. Transgenic cotton, expressing a single Bt toxin, was first used in the 1990s to control H. armigera and other lepidopteran pests. Landscape scale or greater pest suppression has been reported in some countries using this technology. However, a long-term, broad-scale pheromone trapping program for H. armigera in a mixed cropping region in eastern Australia caught more moths during the deployment of single Bt toxin cotton (Ingard®) (1996-2004) than in previous years. This response can be attributed, at least in part, to (1) a precautionary cap (30% of total cotton grown, by area) being applied to Ingard® to restrict the development of Bt resistance in the pest, and (2) during the Ingard® era, cotton production greatly increased (as did that of another host plant, sorghum) and H. armigera (in particular the 3rd and older generations) responded in concert with this increase in host plant availability. However, with the replacement of Ingard® with Bollgard II® cotton (containing two different Bt toxins) in 2005, and recovery of the cotton industry from prevailing drought, H. armigera failed to track increased host-plant supply and moth numbers decreased. Greater toxicity of the two gene product, introduction of no cap on Bt cotton proportion, and an increase in natural enemy abundance are suggested as the most likely mechanisms responsible for the suppression observed.

  9. Identification and validation of reference genes for normalization of gene expression analysis using qRT-PCR in Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Zhang, Songdou; An, Shiheng; Li, Zhen; Wu, Fengming; Yang, Qingpo; Liu, Yichen; Cao, Jinjun; Zhang, Huaijiang; Zhang, Qingwen; Liu, Xiaoxia

    2015-01-25

    Recent studies have focused on determining functional genes and microRNAs in the pest Helicoverpa armigera (Lepidoptera: Noctuidae). Most of these studies used quantitative real-time PCR (qRT-PCR). Suitable reference genes are necessary to normalize gene expression data of qRT-PCR. However, a comprehensive study on the reference genes in H. armigera remains lacking. Twelve candidate reference genes of H. armigera were selected and evaluated for their expression stability under different biotic and abiotic conditions. The comprehensive stability ranking of candidate reference genes was recommended by RefFinder and the optimal number of reference genes was calculated by geNorm. Two target genes, thioredoxin (TRX) and Cu/Zn superoxide dismutase (SOD), were used to validate the selection of reference genes. Results showed that the most suitable candidate combinations of reference genes were as follows: 28S and RPS15 for developmental stages; RPS15 and RPL13 for larvae tissues; EF and RPL27 for adult tissues; GAPDH, RPL27, and β-TUB for nuclear polyhedrosis virus infection; RPS15 and RPL32 for insecticide treatment; RPS15 and RPL27 for temperature treatment; and RPL32, RPS15, and RPL27 for all samples. This study not only establishes an accurate method for normalizing qRT-PCR data in H. armigera but also serve as a reference for further study on gene transcription in H. armigera and other insects. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Isotopes and Trace Elements as Natal Origin Markers of Helicoverpa armigera – An Experimental Model for Biosecurity Pests

    PubMed Central

    Holder, Peter W.; Armstrong, Karen; Van Hale, Robert; Millet, Marc-Alban; Frew, Russell; Clough, Timothy J.; Baker, Joel A.

    2014-01-01

    Protecting a nation's primary production sector and natural estate is heavily dependent on the ability to determine the risk presented by incursions of exotic insect species. Identifying the geographic origin of such biosecurity breaches can be crucial in determining this risk and directing the appropriate operational responses and eradication campaigns, as well as ascertaining incursion pathways. Reading natural abundance biogeochemical markers using mass spectrometry is a powerful tool for tracing ecological pathways as well as provenance determination of commercial products and items of forensic interest. However, application of these methods to trace insects has been underutilised to date and our understanding in this field is still in a phase of basic development. In addition, biogeochemical markers have never been considered in the atypical situation of a biosecurity incursion, where sample sizes are often small, and of unknown geographic origin and plant host. These constraints effectively confound the interpretation of the one or two isotope geo-location markers systems that are currently used, which are therefore unlikely to achieve the level of provenance resolution required in biosecurity interceptions. Here, a novel approach is taken to evaluate the potential for provenance resolution of insect samples through multiple biogeochemical markers. The international pest, Helicoverpa armigera, has been used as a model species to assess the validity of using naturally occurring δ2H, 87Sr/86Sr, 207Pb/206Pb and 208Pb/206Pb isotope ratios and trace element concentration signatures from single moth specimens for regional assignment to natal origin. None of the biogeochemical markers selected were individually able to separate moths from the different experimental regions (150–3000 km apart). Conversely, using multivariate analysis, the region of origin was correctly identified for approximately 75% of individual H. armigera samples. The geographic resolution

  11. Resistance to Bacillus thuringiensis toxin Cry2Ab in a strain of Helicoverpa armigera (Lepidoptera: Noctuidae) in Australia.

    PubMed

    Mahon, R J; Olsen, K M; Garsia, K A; Young, S R

    2007-06-01

    Transgenic cotton, Gossypium hirsutum L., expressing the crylAc and cry2Ab genes from Bacillus thuringiensis (Bt) Berliner variety kurstaki in a pyramid (Bollgard II) was widely planted for the first time in Australia during the 2004-2005 growing season. Before the first commercial Bollgard II crops, limited amounts of cotton expressing only the crylAc gene (Ingard) was grown for seven seasons. No field failures due to resistance to CrylAc toxin were observed during that period and a monitoring program indicated that the frequency of genes conferring high level resistance to the CrylAc toxin were rare in the major pest of cotton, Helicoverpa armigera (Htibner) (Lepidoptera: Noctuidae). Before the deployment of Bollgard II, an allele conferring resistance to Cry2Ab toxin was detected in field-collected H. armigera. We established a colony (designated SP15) consisting of homozygous resistant individuals and examined their characteristics through comparison with individuals from a Bt-susceptible laboratory colony (GR). Through specific crosses and bioassays, we established that the resistance present in SP15 was due to a single autosomal gene. The resistance was recessive. Homozygotes were highly resistant to Cry2Ab toxin, so much so, that we were unable to induce significant mortality at the maximum concentration of toxin available. Homozygotes also were unaffected when fed leaves of a cotton variety expressing the cry2Ab gene. Although cross-resistant to Cry2Aa toxin, SP15 was susceptible to CrylAc and to the Bt product DiPel.

  12. Next-Generation Sequencing-Based Transcriptome Analysis of Helicoverpa armigera Larvae Immune-Primed with Photorhabdus luminescens TT01

    PubMed Central

    Zhao, Zengyang; Wu, Gongqing; Wang, Jia; Liu, Chunlin; Qiu, Lihong

    2013-01-01

    Although invertebrates are incapable of adaptive immunity, immunal reactions which are functionally similar to the adaptive immunity of vertebrates have been described in many studies of invertebrates including insects. The phenomenon was termed immune priming. In order to understand the molecular mechanism of immune priming, we employed Illumina/Solexa platform to investigate the transcriptional changes of the hemocytes and fat body of Helicoverpa armigera larvae immune-primed with the pathogenic bacteria Photorhabdus luminescens TT01. A total of 43.6 and 65.1 million clean reads with 4.4 and 6.5 gigabase sequence data were obtained from the TT01 (the immune-primed) and PBS (non-primed) cDNA libraries and assembled into 35,707 all-unigenes (non-redundant transcripts), which has a length varied from 201 to 16,947 bp and a N50 length of 1,997 bp. For 35,707 all-unigenes, 20,438 were functionally annotated and 2,494 were differentially expressed after immune priming. The differentially expressed genes (DEGs) are mainly related to immunity, detoxification, development and metabolism of the host insect. Analysis on the annotated immune related DEGs supported a hypothesis that we proposed previously: the immune priming phenomenon observed in H. armigera larvae was achieved by regulation of key innate immune elements. The transcriptome profiling data sets (especially the sequences of 1,022 unannotated DEGs) and the clues (such as those on immune-related signal and regulatory pathways) obtained from this study will facilitate immune-related novel gene discovery and provide valuable information for further exploring the molecular mechanism of immune priming of invertebrates. All these will increase our understanding of invertebrate immunity which may provide new approaches to control insect pests or prevent epidemic of infectious diseases in economic invertebrates in the future. PMID:24302999

  13. Infection of Helicoverpa armigera by endophytic Beauveria bassiana colonizing tomato plants

    USDA-ARS?s Scientific Manuscript database

    A novel endophytic strain of Beauveria bassiana was isolated from leaf tissue of a wild tomato plant. This strain and two B. bassiana strains previously isolated from soil were evaluated for their ability to endophytically colonize tomatoes and subsequent in planta efficacy against Helicoverpa armig...

  14. Development associated profiling of chitinase and microRNA of Helicoverpa armigera identified chitinase repressive microRNA.

    PubMed

    Agrawal, Neema; Sachdev, Bindiya; Rodrigues, Janneth; Sree, K Sowjanya; Bhatnagar, Raj K

    2013-01-01

    Expression of chitinase is developmentally regulated in insects in consonance with their molting process. During the larval-larval metamorphosis in Helicoverpa armigera, chitinase gene expression varies from high to negligible. In the five-day metamorphic course of fifth-instar larvae, chitinase transcript is least abundant on third day and maximal on fifth day. MicroRNA library prepared from these highest and lowest chitinase-expressing larval stages resulted in isolation of several miRNAs. In silico analysis of sequenced miRNAs revealed three miRNAs having sequence similarity to 3'UTR of chitinase. Gene-targeted specific action of these miRNAs, was investigated by luciferase reporter having 3'UTR of chitinase. Only one of three miRNAs, miR-24, inhibited luciferase expression. Further, a day-wise in vivo quantification of miR-24 in fifth-instar larvae revealed a negative correlation with corresponding chitinase transcript abundance. The force-feeding of synthetic miR-24 induced significant morphological aberrations accompanied with arrest of molting. These miR-24 force-fed larvae revealed significantly reduced chitinase transcript abundance.

  15. Dynamic transcriptome analysis and volatile profiling of Gossypium hirsutum in response to the cotton bollworm Helicoverpa armigera

    PubMed Central

    Huang, Xin-Zheng; Chen, Jie-Yin; Xiao, Hai-Jun; Xiao, Yu-Tao; Wu, Juan; Wu, Jun-Xiang; Zhou, Jing-Jiang; Zhang, Yong-Jun; Guo, Yu-Yuan

    2015-01-01

    In response to insect herbivory, plants emit elevated levels of volatile organic compounds for direct and indirect resistance. However, little is known about the molecular and genomic basis of defense response that insect herbivory trigger in cotton plants and how defense mechanisms are orchestrated in the context of other biological processes. Here we monitored the transcriptome changes and volatile characteristics of cotton plants in response to cotton bollworm (CBW; Helicoverpa armigera) larvae infestation. Analysis of samples revealed that 1,969 transcripts were differentially expressed (log2|Ratio| ≥ 2; q ≤ 0.05) after CBW infestation. Cluster analysis identified several distinct temporal patterns of transcriptome changes. Among CBW-induced genes, those associated with indirect defense and jasmonic acid pathway were clearly over-represented, indicating that these genes play important roles in CBW-induced defenses. The gas chromatography-mass spectrometry (GC-MS) analyses revealed that CBW infestation could induce cotton plants to release volatile compounds comprised lipoxygenase-derived green leaf volatiles and a number of terpenoid volatiles. Responding to CBW larvae infestation, cotton plants undergo drastic reprogramming of the transcriptome and the volatile profile. The present results increase our knowledge about insect herbivory-induced metabolic and biochemical processes in plants, which may help improve future studies on genes governing processes. PMID:26148847

  16. Dynamic transcriptome analysis and volatile profiling of Gossypium hirsutum in response to the cotton bollworm Helicoverpa armigera.

    PubMed

    Huang, Xin-Zheng; Chen, Jie-Yin; Xiao, Hai-Jun; Xiao, Yu-Tao; Wu, Juan; Wu, Jun-Xiang; Zhou, Jing-Jiang; Zhang, Yong-Jun; Guo, Yu-Yuan

    2015-07-07

    In response to insect herbivory, plants emit elevated levels of volatile organic compounds for direct and indirect resistance. However, little is known about the molecular and genomic basis of defense response that insect herbivory trigger in cotton plants and how defense mechanisms are orchestrated in the context of other biological processes. Here we monitored the transcriptome changes and volatile characteristics of cotton plants in response to cotton bollworm (CBW; Helicoverpa armigera) larvae infestation. Analysis of samples revealed that 1,969 transcripts were differentially expressed (log2|Ratio| ≥ 2; q ≤ 0.05) after CBW infestation. Cluster analysis identified several distinct temporal patterns of transcriptome changes. Among CBW-induced genes, those associated with indirect defense and jasmonic acid pathway were clearly over-represented, indicating that these genes play important roles in CBW-induced defenses. The gas chromatography-mass spectrometry (GC-MS) analyses revealed that CBW infestation could induce cotton plants to release volatile compounds comprised lipoxygenase-derived green leaf volatiles and a number of terpenoid volatiles. Responding to CBW larvae infestation, cotton plants undergo drastic reprogramming of the transcriptome and the volatile profile. The present results increase our knowledge about insect herbivory-induced metabolic and biochemical processes in plants, which may help improve future studies on genes governing processes.

  17. Molecular and biochemical characterization of the effects of insecticidal toxin from Meloidae beetles on Helicoverpa armigera (Hub.) (Lepidoptera: Noctuidae).

    PubMed

    Khan, R A; Rashid, M; Wang, D; Zhang, Y L

    2013-10-10

    The molecular and biochemical effects of an insecticidal toxin extracted from Meloidae beetles were investigated on Helicoverpa armigera. The toxin was identified as cantharidin, a well-known natural compound produced by beetles of family Meloidae and Oedemeridae. Furthermore, the effect of the toxin on the metabolic enzymes alkaline phosphatase (ALP) and glutathione S-transferase (GST), responsible for the metabolism of insecticides, was also investigated. Results of a diet incorporation bioassay performed under laboratory conditions showed that the LC50 value of cantharidin was 0.068 mg/g. The body weight of the insect was also significantly reduced by cantharidin treatment. The LC10 concentration of cantharidin, 0.01 mg/g, was also tested to determine its effect on ALP and GST. Our results showed that cantharidin significantly inhibited ALP activity after 48 h, whereas GST activity was significantly inhibited after 24 h. The decline of ALP and GST transcript levels was also validated by semiquantitative RT-PCR analysis. It may be concluded from the results that ALPs and GSTs may be targets of the cantharidin intoxication mechanism. Moreover, the inability of ALP and GST to metabolize cantharidin shows that the mechanism of detoxification for cantharidin is different from that for conventional insecticides. On the basis of our investigations, the chemical structure of insecticides may be modified using a model structure of cantharidin, to avoid metabolism by metabolic enzymes.

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

    PubMed

    Liu, D; Yuan, Y; Li, M; Qiu, X

    2015-12-01

    Quercetin 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.

  19. Functional analysis of the ABCs of eye color in Helicoverpa armigera with CRISPR/Cas9-induced mutations

    PubMed Central

    Khan, Sher Afzal; Reichelt, Michael; Heckel, David G.

    2017-01-01

    Many insect pigments are localized in subcellular pigment granules, and transport of pigment precursors from the cytoplasm is accomplished by ABC proteins. Drosophila melanogaster has three half-transporter genes (white, scarlet, and brown, all affecting eye pigments) and Bombyx mori has a fourth (ok). The White, Brown, Scarlet and Ok proteins each have one transmembrane and one cytoplasmic domain and they heterodimerize to form functional transporters with different substrate specificities. We used CRISPR/Cas9 to create somatic and germ-line knockout mutations of these four genes in the noctuid moth Helicoverpa armigera. Somatic knockouts of white block pigmentation of the egg, first instar larva and adult eye, but germ-line knockouts of white are recessive lethal in the embryo. Knockouts of scarlet are viable and produce pigmentless first instar larvae and yellow adult eyes lacking xanthommatin. Knockouts of brown show no phenotypic effects on viability or pigmentation. Knockouts of ok are viable and produce translucent larval cuticle and black eyes. CRISPR/Cas9-induced mutations are a useful tool for analyzing how essential and non-essential genes interact to produce the diversity of insect pigmentation patterns found in nature. PMID:28053351

  20. Daily expression of two circadian clock genes in compound eye of Helicoverpa armigera: evidence for peripheral tissue circadian timing.

    PubMed

    Yan, Shuo; Liu, Yan-Jun; Zhu, Jia-Lin; Cui, Wei-Na; Zhang, Xin-Fang; Yang, Yu-Hui; Liu, Xiao-Ming; Zhang, Qing-Wen; Liu, Xiao-Xia

    2017-09-23

    Circadian clock genes in peripheral tissues usually play an important role in regulating the circadian rhythms. Light is the most important environmental signal for synchronizing endogenous rhythms with the daily light-dark cycle, and compound eyes are known as the principal circadian photoreceptor for photic entrainment in most moths. However, there is little evidence for circadian timing in the compound eyes. In the current study, we isolated the timeless gene, designated Ha-tim (GenBank accession number: KM233162), from the cotton bollworm Helicoverpa armigera. Ha-tim and period (Ha-per) showed low mRNA levels in the compound eyes compared to the other tested adult organs. Ha-tim and Ha-per transcript levels were dependent on an endogenous rhythm that fluctuated over a daily cycle in the compound eyes and heads. The cycles of Ha-tim and Ha-per transcript levels followed similar time courses, and identical expression patterns of the two genes were observed in the compound eyes and heads. Ha-tim and Ha-per were down-regulated in the compound eyes after light exposure, copulation and starvation. These results indicated that Ha-tim and Ha-per transcript levels were regulated by endogenous and exogenous factors. Our study helped to improve our understanding of the circadian clock machinery in compound eyes and other peripheral tissues. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  1. A global-wide search for sexual dimorphism of glomeruli in the antennal lobe of female and male Helicoverpa armigera

    PubMed Central

    Zhao, Xin-Cheng; Ma, Bai-Wei; Berg, Bente G.; Xie, Gui-Ying; Tang, Qing-Bo; Guo, Xian-Ru

    2016-01-01

    By using immunostaining and three-dimensional reconstruction, the anatomical organization of the antennal lobe glomeruli of the female cotton bollworm Helicoverpa armigera was investigated. Eighty-one glomeruli were identified, 15 of which were not previously discovered. The general anatomical organization of the AL of female is similar to that of male and all glomeruli were classified into four sub-groups, including the female-specific glomerular complex, posterior complex, labial-palp pit organ glomerulus, and ordinary glomeruli. A global-wide comparison on the complete glomerular map of female and male was performed and for the first time the quantitative difference in volume for each individual homologous glomerulus was analyzed. We found that the sexual dimorphism includes not only the sex-specific glomeruli but also some of the other glomeruli. The findings in the present study may provide a reference to examine the antennal-lobe organization more in detail and to identify new glomeruli in other moth species. In addition, the complete identification and global-wide comparison of the sexes provide an important basis for mapping the function of distinct glomeruli and for understanding neural mechanisms underlying sexually dimorphic olfactory behaviors. PMID:27725758

  2. Molecular cloning and expression patterns of the molt-regulating transcription factor HHR3 from Helicoverpa armigera.

    PubMed

    Zhao, X-F; Wang, J-X; Xu, X-L; Li, Z-M; Kang, C-J

    2004-08-01

    Molt-regulating transcription factors, hormone receptor 3 (HR3), play important roles in regulating expression of tissue-specific genes involved in insect molting and metamorphosis. A 1668 bp cDNA encoding a molt-regulating transcription factor (HHR3) was cloned from Helicoverpa armigera, which encodes a protein made up of 556 amino acids. This 62 kDa protein was found to have an isoelectric point (pI) of 6.52. There was no signal peptide or N-glycosylation site found in this cDNA. A DNA-binding region signature of nuclear hormone receptor was found from amino acids 107-133. A possible outside to inside transmembrane helice was found from amino acids 72-90. Northern blots of the larvae revealed five bands of HHR3 named as band 0, 1, 2, 3 and 4 with molecular masses determined as 2.1, 2.6, 3.6, 4.5 and 5.5 kb, respectively. The expression patterns of HHR3 in vivo were variable with developmental stages and tissues. Results showed that band 1-4 of HHR3 was only briefly expressed during molting, which suggested these bands are involved in the regulation of molting cascade, whereas band 0 was expressed in both molting and feeding larvae. Band 1 and 2 of HHR3 could be induced from epidermis of newly molted 6th instar larvae by non-steroidal ecdysone agonist, RH-2485.

  3. Central projections of gustatory receptor neurons in the medial and the lateral sensilla styloconica of Helicoverpa armigera larvae.

    PubMed

    Tang, Qing-Bo; Zhan, Huan; Cao, Huan; Berg, Bente G; Yan, Feng-Ming; Zhao, Xin-Cheng

    2014-01-01

    Food selection behavior of lepidopteran larvae is predominantly governed by the activation of taste neurons present in two sensilla styloconica located on the galea of the maxilla. In this study, we present the ultrastructure of the sensilla styloconica and the central projection pattern of their associated receptor neurons in larvae of the heliothine moth, Helicoverpa armigera. By means of light microscopy and scanning electron microscopy, the previous findings of two morphologically fairly similar sensilla comprising a socketed conic tip inserted into a large peg were confirmed. However, the peg size of the medial sensillum was found to be significantly bigger than that of the lateral sensillum. The sensory neurons derived from each sensillum styloconicum were mapped separately using anterograde staining experiments combined with confocal laser-scanning microscopy. For determining the afferents' target regions relative to each other, we reconstructed the labeled axons and placed them into a common reference framework. The sensory axons from both sensilla projected via the ipsilateral maxillary nerve to the suboesophageal ganglion and further through the ipsilateral circumoesophageal connective to the brain. In the suboesophageal ganglion, the sensory projections targeted two areas of the ipsilateral maxillary neuropil, one located in the ventrolateral neuromere and the other adjacent to the neuromere midline. In the brain, the axon terminals targeted the dorso-anterior area of the ipsilateral tritocerebrum. As confirmed by the three-dimensional reconstructions, the target regions of the neural projections originating from each of the two sensilla styloconica were identical.

  4. Nutritional performance and activity of some digestive enzymes of the cotton bollworm, Helicoverpa armigera, in response to seven tested bean cultivars.

    PubMed

    Namin, Foroogh Rahimi; Naseri, Bahram; Razmjou, Jabraeil

    2014-01-01

    Nutritional performance and activity of some digestive enzymes (protease and α-amylase) of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) in response to feeding on bean (Phaseolus vulgaris L. (Fabales: Fabaceae)) cultivars (Shokufa, Akhtar, Sayyad, Naz, Pak, Daneshkadeh, and Talash) were evaluated under laboratory conditions (25 ± 1°C, 65 ± 5% RH, and a 16:8 L:D photoperiod). The highest and lowest respective values of approximate digestibility were observed when fourth, fifth, and sixth larval instar H. armigera were fed red kidney bean Akhtar and white kidney bean Daneshkadeh. The efficiency of conversion of ingested and digested food was highest when H. armigera was fed red kidney beans Akhtar and Naz and lowest when they were fed white kidney bean Pak. The highest protease activity of fifth instars was observed when they were fed red kidney bean Naz, and the highest amylase activity of fifth instars was observed when they were fed red kidney bean Sayyad. Sixth instar larvae that fed on red kidney bean Sayyad showed the highest protease activity. Larvae reared on common bean Talash and white kidney bean Pak showed the highest amylase activity. Among bean cultivars tested, red kidney bean Sayyad was the most unsuitable host for feeding H. armigera.

  5. A Multiplex Real-Time PCR Assay to Diagnose and Separate Helicoverpa armigera and H. zea (Lepidoptera: Noctuidae) in the New World.

    PubMed

    Gilligan, Todd M; Tembrock, Luke R; Farris, Roxanne E; Barr, Norman B; van der Straten, Marja J; van de Vossenberg, Bart T L H; Metz-Verschure, Eveline

    2015-01-01

    The Old World bollworm, Helicoverpa armigera (Hübner), and the corn earworm, H. zea (Boddie), are two of the most important agricultural pests in the world. Diagnosing these two species is difficult-adults can only be separated with a complex dissection, and larvae cannot be identified to species using morphology, necessitating the use of geographic origin for identification in most instances. With the discovery of H. armigera in the New World, identification of immature Helicoverpa based on origin is no longer possible because H. zea also occurs in all of the geographic regions where H. armigera has been discovered. DNA barcoding and restriction fragment length polymorphism (RFLP) analyses have been reported in publications to distinguish these species, but these methods both require post-PCR processing (i.e., DNA sequencing or restriction digestion) to complete. We report the first real-time PCR assay to distinguish these pests based on two hydrolysis probes that bind to a segment of the internal transcribed spacer region 2 (ITS2) amplified using a single primer pair. One probe targets H. armigera, the second probe targets H. zea, and a third probe that targets a conserved segment of 18S rDNA is used as a control of DNA quality. The assay can be completed in 50 minutes when using isolated DNA and is successfully tested on larvae intercepted at ports of entry and adults captured during domestic surveys. We demonstrate that the assay can be run in triplex with no negative effects on sensitivity, can be run using alternative real-time PCR reagents and instruments, and does not cross react with other New World Heliothinae.

  6. A Multiplex Real-Time PCR Assay to Diagnose and Separate Helicoverpa armigera and H. zea (Lepidoptera: Noctuidae) in the New World

    PubMed Central

    Gilligan, Todd M.; Tembrock, Luke R.; Farris, Roxanne E.; Barr, Norman B.; van der Straten, Marja J.; van de Vossenberg, Bart T. L. H.; Metz-Verschure, Eveline

    2015-01-01

    The Old World bollworm, Helicoverpa armigera (Hübner), and the corn earworm, H. zea (Boddie), are two of the most important agricultural pests in the world. Diagnosing these two species is difficult—adults can only be separated with a complex dissection, and larvae cannot be identified to species using morphology, necessitating the use of geographic origin for identification in most instances. With the discovery of H. armigera in the New World, identification of immature Helicoverpa based on origin is no longer possible because H. zea also occurs in all of the geographic regions where H. armigera has been discovered. DNA barcoding and restriction fragment length polymorphism (RFLP) analyses have been reported in publications to distinguish these species, but these methods both require post-PCR processing (i.e., DNA sequencing or restriction digestion) to complete. We report the first real-time PCR assay to distinguish these pests based on two hydrolysis probes that bind to a segment of the internal transcribed spacer region 2 (ITS2) amplified using a single primer pair. One probe targets H. armigera, the second probe targets H. zea, and a third probe that targets a conserved segment of 18S rDNA is used as a control of DNA quality. The assay can be completed in 50 minutes when using isolated DNA and is successfully tested on larvae intercepted at ports of entry and adults captured during domestic surveys. We demonstrate that the assay can be run in triplex with no negative effects on sensitivity, can be run using alternative real-time PCR reagents and instruments, and does not cross react with other New World Heliothinae. PMID:26558366

  7. Transcriptional responses underlying the hormetic and detrimental effects of the plant secondary metabolite gossypol on the generalist herbivore Helicoverpa armigera

    PubMed Central

    2011-01-01

    Background Hormesis is a biphasic biological response characterized by the stimulatory effect at relatively low amounts of chemical compounds which are otherwise detrimental at higher concentrations. A hormetic response in larval growth rates has been observed in cotton-feeding insects in response to increasing concentrations of gossypol, a toxic metabolite found in the pigment glands of some plants in the family Malvaceae. We investigated the developmental effect of gossypol in the cotton bollworm, Helicoverpa armigera, an important heliothine pest species, by exposing larvae to different doses of this metabolite in their diet. In addition, we sought to determine the underlying transcriptional responses to different gossypol doses. Results Larval weight gain, pupal weight and larval development time were measured in feeding experiments and a hormetic response was seen for the first two characters. On the basis of net larval weight gain responses to gossypol, three concentrations (0%, 0.016% and 0.16%) were selected for transcript profiling in the gut and the rest of the body in a two-color double reference design microarray experiment. Hormesis could be observed at the transcript level, since at the low gossypol dose, genes involved in energy acquisition such as β-fructofuranosidases were up-regulated in the gut, and genes involved in cell adhesion were down-regulated in the body. Genes with products predicted to be integral to the membrane or associated with the proteasome core complex were significantly affected by the detrimental dose treatment in the body. Oxidoreductase activity-related genes were observed to be significantly altered in both tissues at the highest gossypol dose. Conclusions This study represents the first transcriptional profiling approach investigating the effects of different concentrations of gossypol in a lepidopteran species. H. armigera's transcriptional response to gossypol feeding is tissue- and dose-dependent and involves diverse

  8. Selection and heritability of resistance to Bacillus thuringiensis subsp kurstaki and transgenic cotton in Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Lu, Mei-guang; Rui, Chang-Hui; Zhao, Jian-Zhou; Jian, Gui-liang; Fan, Xian-lin; Gao, Xi-wu

    2004-09-01

    Compared with an unselected susceptible population, a cotton bollworm, Helicoverpa armigera (Hübner), population selected for 22 generations with transgenic cotton leaves (modified Cry1A) in the laboratory developed 11.0-fold resistance to Cry1Ac (one single-protein product MVPII). Resistance to Bacillus thuringiensis Berliner subsp kurstaki (Btk) was selected for 22 generations with a 5.2-fold increase in LC50. The estimated realized heritabilities (h2) of resistance for transgenic-cotton- and Btk-selected populations were 0.1008 and 0.2341, respectively. This reflects the higher phenotypic variation in response to Cry1Ac in the transgenic-cotton-selected population. This variation may have been caused by differences in protein toxin levels expressed in different growth stages of the transgenic cotton. Because of the different slopes of the probit regression lines between Cry1Ac and Btk, the estimated realized h2 cannot be used visually to compare resistance development to Cry1Ac and Btk in H armigera. Thus, the response quotient (Q) of resistance was also estimated. The Q values of resistance for transgenic-cotton- and Btk-selected populations were 0.0763 and 0.0836, respectively. This showed that the rate of resistance development would be similar in both selection populations. This result indicates that the selection of resistance using transgenic cotton is different from that selected using the single toxin. Resistance risk to transgenic cotton and Btk in field populations was assessed assuming different pressures of selection by using the estimated h2. Assuming the h2 of resistance in a field population was half of the estimated h2, and the population received prolonged and uniform exposure to transgenic cotton or Btk causing >70% mortality in each generation, we predicted that resistance would increase 10-fold after <23 generations for Cry1Ac in transgenic cotton-selected-populations and after <21 generations for Btk in Btk-selected populations. Cross

  9. The seesaw effect of winter temperature change on the recruitment of cotton bollworms Helicoverpa armigera through mismatched phenology.

    PubMed

    Reddy, Gadi V P; Shi, Peijian; Hui, Cang; Cheng, Xiaofei; Ouyang, Fang; Ge, Feng

    2015-12-01

    Knowing how climate change affects the population dynamics of insect pests is critical for the future of integrated pest management. Rising winter temperatures from global warming can drive increases in outbreaks of some agricultural pests. In contrast, here we propose an alternative hypothesis that both extremely cold and warm winters can mismatch the timing between the eclosion of overwintering pests and the flowering of key host plants. As host plants normally need higher effective cumulative temperatures for flowering than insects need for eclosion, changes in flowering time will be less dramatic than changes in eclosion time, leading to a mismatch of phenology on either side of the optimal winter temperature. We term this the "seesaw effect." Using a long-term dataset of the Old World cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in northern China, we tested this seesaw hypothesis by running a generalized additive model for the effects of the third generation moth in the preceding year, the winter air temperature, the number of winter days below a critical temperature and cumulative precipitation during winter on the demography of the overwintering moth. Results confirmed the existence of the seesaw effect of winter temperature change on overwintering populations. Pest management should therefore consider the indirect effect of changing crop phenology (whether due to greenhouse cultivation or to climate change) on pest outbreaks. As arthropods from mid- and high latitudes are actually living in a cooler thermal environment than their physiological optimum in contrast to species from lower latitudes, the effects of rising winter temperatures on the population dynamics of arthropods in the different latitudinal zones should be considered separately. The seesaw effect makes it more difficult to predict the average long-term population dynamics of insect pests at high latitudes due to the potential sharp changes in annual growth rates

  10. Procarboxypeptidase A from the insect pest Helicoverpa armigera and its derived enzyme. Two forms with new functional properties.

    PubMed

    Bayés, Alex; Sonnenschein, Anka; Daura, Xavier; Vendrell, Josep; Aviles, Francesc X

    2003-07-01

    Although there is a significant knowledge about mammalian metallocarboxypeptidases, the data available on this family of enzymes is very poor for invertebrate forms. Here we present the biochemical characterization of a metallocarboxypeptidase from the insect Helicoverpa armigera (Lepidoptera: Noctuidae), a devastating pest spread in subtropical regions of Europe, Asia, Africa and Oceania. The zymogen of this carboxypeptidase (PCPAHa) has been expressed at high levels in a Pichia pastoris system and shown to display the characteristics of the enzyme purified from the insect midgut. The in vitro activation process of the proenzyme differs significantly from the mammalian ones. The lysine-specific endoprotease LysC activates PCPAHa four times more efficiently than trypsin, the general activating enzyme for all previously studied metalloprocarboxypeptidases. LysC and trypsin independently use two different activation targets and the presence of sugars in the vicinity of the LysC activation point affects the activation process, indicating a possible modulation of the activation mechanism. During the activation with LysC the prodomain is degraded, while the carboxypeptidase moiety remains intact except for a C-terminal octapeptide that is rapidly released. Interestingly, the sequence at the cleavage point for the release of the octapeptide is also found at the boundary between the activation peptide and the enzyme moieties. The active enzyme (CPAHa) is shown to have a very broad substrate specificity, as it appears to be the only known metallocarboxypeptidase capable of efficiently hydrolysing basic and aliphatic residues and, to a much lower extent, acidic residues. Two carboxypeptidase inhibitors, from potato and leech, were tested against CPAHa. The former, of vegetal origin, is the most efficient metallocarboxypeptidase inhibitor described so far, with a Ki in the pm range.

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

    PubMed Central

    Xiao, Haijun; Xie, Bingtang; Smagghe, Guy; Guo, Yuyuan; Liang, Gemei

    2016-01-01

    Developing oocytes accumulate plentiful yolk protein during oogenesis through receptor-mediated endocytosis. The vitellogenin receptor (VgR), belonging to the low-density lipoprotein receptor (LDLR) family, regulates the absorption of yolk protein. In this work, the full-length vitellogenin receptor (HaVgR) in the cotton bollworm Helicoverpa armigera was identified, encoding a 1817 residue protein. Sequence alignment revealed that the sequence of HaVgR contained all of the conservative structural motifs of LDLR family members, and phylogenetic analysis indicated that HaVgR had a high identity among Lepidoptera and was distinct from that of other insects. Consistent with other insects, HaVgR was specifically expressed in ovarian tissue. The developmental expression pattern showed that HaVgR was first transcribed in the newly metamorphosed female adults, reached a peak in 2-day-old adults and then declined. Western blot analysis also revealed an ovarian-specific and developing expression pattern, which was consistent with the HaVgR mRNA transcription. Moreover, RNAi-mediated HaVgR knockdown strongly reduced the VgR expression in both the mRNA and protein levels, which inhibited the yolk protein deposition in the ovaries, led to the dramatic accumulation of vitellogenin and the up-regulation of HaVg expression in hemolymph, and eventually resulted in a declined fecundity. Together, all of these findings demonstrate that HaVgR is a specific receptor in uptake and transportation of yolk protein for the maturation of oocytes and that it plays a critical role in female reproduction. PMID:27192057

  12. The retardant effect of 2-Tridecanone, mediated by Cytochrome P450, on the Development of Cotton bollworm, Helicoverpa armigera.

    PubMed

    Zhang, Lei; Lu, Yao; Xiang, Min; Shang, Qingli; Gao, Xiwu

    2016-11-22

    Plant allelochemicals act as toxins, inhibitors of digestion, and deterrents that affect the fecundity of insects. These compounds have attracted significant research attention in recent decades, and much is known about the effects of these xenobiotic plant secondary metabolites on insect development. To date, although ecological interactions between xenobiotic plant secondary chemicals that retard insect growth have been observed in many species, it remains unclear how particular allelochemicals influence insect development in a life stage-dependent manner. We found that 2-tridecanone can affect insect development; this effect appears similar to the symptoms induced by the physiological imbalance between juvenile and molting hormones in cotton bollworm. We later detected that a decrease in the concentration of 20-hydroxyecdysone occurred alongside the observed symptoms. We next identified the transcriptome of Helicoverpa armigera and eightdigital gene expression libraries for shading light on how 2-tridecanone retarded the development of cotton bollworm. The expression of CYP314A1, CYP315A1, CYP18A1, CYP307A1, and CYP306A1 (unigenes 16487, 15409, 40026, 41217, 35643, 16953, 8199, 13311, and 13036) were found to be induced by 2-tridecanone; these are known to be related to the biosynthesis or metabolism of 20-hydroxyecdysone. Expression analysis and RNA interference studies established that the retardant effect of 2-tridecanone on the development of cotton bollworm is mediated by P450 genes. The candidate P450 gene approach described and exploited here is useful for identifying potential causal genes for the influence of plant allelochemicals on insect development.

  13. A comparison of the life-history traits between diapause and direct development individuals in the cotton bollworm, Helicoverpa armigera.

    PubMed

    Chen, Chao; Xia, Qin-Wen; Xiao, Hai-Jun; Xiao, Liang; Xue, Fang-Sen

    2014-02-05

    In order to understand the differences of life-history traits between diapause and direct development individuals in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), the development time, body size, growth rate, and adult longevity were investigated between the two populations, which were induced under 12:12 L:D and 16:8 L:D photoperiods, respectively, at 20, 22, and 25°C. The results indicated that the larval development time, pupal weight, adult weight, and growth rate were significantly different between diapause and direct developing individuals. The diapause developing individuals had a significantly higher pupal and adult weight and a longer larval time compared with direct developing individuals. However, the growth rate in diapause developing individuals was lower than that in the direct developing individuals. Analysis by GLM showed that larval time, pupal and adult weight, and growth rate were significantly influenced by both temperature and developmental pathway. The pupal and adult weights were greater in males than females in both developmental pathways, exhibiting sexual size dimorphism. The dimorphism in adult weight was more pronounced than in pupal weight because female pupae lost more weight at metamorphosis compared to male pupae. Protogyny was observed in both developmental pathways. However, the protogyny phenomenon was more pronounced at lower temperatures in direct developing individuals, whereas it was more pronounced in diapause developing individuals when they experienced higher temperatures in their larval stage and partial pupal period. The adult longevity of diapause developing individuals was significantly longer than that of direct developing individuals. The results reveal that the life-history strategy was different between diapause and direct developing individuals. This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper

  14. Inheritance of Cry1Ac resistance and associated biological traits in the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Nair, Rupa; Kalia, Vinay; Aggarwal, K K; Gujar, Govind T

    2010-05-01

    The analysis of reciprocal genetic crosses between resistant Helicoverpa armigera strain (BH-R) (227.9-fold) with susceptible Vadodara (VA-S) strain showed dominance (h) of 0.65-0.89 and degree of dominance (D) of 0.299-0.782 suggesting Cry1Ac resistance as a semi-dominant trait. The D and h values of F(1) hybrids of female resistant parent were higher than female susceptible parent, showing maternally enhanced dominance of Cry1Ac resistance. The progeny of F(2) crosses, backcrosses of F(1) hybrid with resistant BH-R parent did not differ significantly in respect of mortality response with resistant parent except for backcross with female BH-R and male of F(1) (BH-RxVA-S) cross, suggesting dominant inheritance of Cry1Ac resistance. Evaluation of some biological attributes showed that larval and pupal periods of progenies of reciprocal F(1) crosses, backcrosses and F(2) crosses were either at par with resistant parent or lower than susceptible parent on treated diet (0.01 microg/g). The susceptible strain performed better in terms of pupation and adult formation than the resistant strain on untreated diet. In many backcrosses and F(2) crosses, Cry1Ac resistance favored emergence of more females than males on untreated diet. The normal larval period and the body weight (normal larval growth) were the dominant traits associated with susceptible strain as contrast to longer larval period and the lower body weight (slow growth) associated with resistance trait. Further, inheritance of larval period in F(2) and backcross progeny suggested existence of a major resistant gene or a set of tightly linked loci associated with Cry1Ac sensitivity. (c) 2010 Elsevier Inc. All rights reserved.

  15. Wnt/β-catenin signaling regulates Helicoverpa armigera pupal development by up-regulating c-Myc and AP-4.

    PubMed

    Chen, Wei; Xu, Wei-Hua

    2014-10-01

    Seasonally changing environmental conditions perceived by insect brains can be converted into hormonal signals that prompt insects to make a decision to develop or enter developmental arrest (diapause). Diapause is a complex physiological response, and many signaling pathways may participate in its regulation. However, little is known about these regulatory pathways. In this study, we cloned four genes related to the Wnt/β-catenin signaling pathway from Helicoverpa armigera, a pupal diapause species. Western blotting shows that expression of Har-Wnt1, Har-β-catenin, and Har-c-Myc are higher in non-diapause pupal brains than in diapause-destined brains. Har-Wnt1 can promote the accumulation of Har-β-catenin in the nucleus, and Har-β-catenin in turn increases the expression of Har-c-Myc. The blockage of Wnt/β-catenin signaling by the inhibitor XAV939 significantly down-regulates Har-β-catenin and Har-c-Myc expression and delays pupal development, suggesting that the Wnt/β-catenin pathway functions in insect development. Furthermore, Har-c-Myc binds to the promoter of Har-AP-4 and regulates its expression. It has been reported that Har-AP-4 activates diapause hormone (DH) expression and that DH up-regulates the growth hormone ecdysteroid for pupal development. Thus, pupal development is regulated by Wnt/β-catenin signaling through the pathway Wnt-β-catenin-c-Myc-AP-4-DH-ecdysteroid. In contrast, the down-regulation of Wnt/β-catenin signaling is likely to induce insects to enter diapause. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Effectiveness of Bacillus thuringiensis-Transgenic Chickpeas and the Entomopathogenic Fungus Metarhizium anisopliae in Controlling Helicoverpa armigera (Lepidoptera: Noctuidae)▿

    PubMed Central

    Lawo, N. C.; Mahon, R. J.; Milner, R. J.; Sarmah, B. K.; Higgins, T. J. V.; Romeis, J.

    2008-01-01

    The use of genetically modified (Bt) crops expressing lepidopteran-specific Cry proteins derived from the soil bacterium Bacillus thuringiensis is an effective method to control the polyphagous pest Helicoverpa armigera. As H. armigera potentially develops resistance to Cry proteins, Bt crops should be regarded as one tool in integrated pest management. Therefore, they should be compatible with biological control. Bioassays were conducted to understand the interactions between a Cry2Aa-expressing chickpea line, either a susceptible or a Cry2A-resistant H. armigera strain, and the entomopathogenic fungus Metarhizium anisopliae. In a first concentration-response assay, Cry2A-resistant larvae were more tolerant of M. anisopliae than susceptible larvae, while in a second bioassay, the fungus caused similar mortalities in the two strains fed control chickpea leaves. Thus, resistance to Cry2A did not cause any fitness costs that became visible as increased susceptibility to the fungus. On Bt chickpea leaves, susceptible H. armigera larvae were more sensitive to M. anisopliae than on control leaves. It appeared that sublethal damage induced by the B. thuringiensis toxin enhanced the effectiveness of M. anisopliae. For Cry2A-resistant larvae, the mortalities caused by the fungus were similar when they were fed either food source. To examine which strain would be more likely to be exposed to the fungus, their movements on control and Bt chickpea plants were compared. Movement did not appear to differ among larvae on Bt or conventional chickpeas, as indicated by the number of leaflets damaged per leaf. The findings suggest that Bt chickpeas and M. anisopliae are compatible to control H. armigera. PMID:18487396

  17. Influence of oxalic and malic acids in chickpea leaf exudates on the biological activity of CryIAc towards Helicoverpa armigera.

    PubMed

    Devi, V Surekha; Sharma, Hari C; Rao, P Arjuna

    2013-04-01

    Efforts are being made to express toxin genes from the bacterium, Bacillus thuringiensis (Bt) in chickpea for minimizing the losses due to the pod borer, Helicoverpa armigera. However, there is an apprehension that acidic exudates in chickpea leaves may influence the protoxin-toxin conversion in the insect midgut, and thus, reduce the efficacy of Bt toxins. Therefore, we studied the influence of organic acids (oxalic acid and malic acid) present in the trichome exudates of chickpea on the biological activity and binding of Bt δ-endotoxin Cry1Ac to brush border membrane vesicles (BBMV) of the pod borer, H. armigera. Oxalic and malic acids in combination at concentrations present in chickpea leaves did not influence the biological activity of Bt toxin Cry1Ac towards H. armigera larvae. Amounts of Cry1Ac protein in the midgut of insects reared on diets with organic acids were similar to those reared on artificial diet without the organic acids. However, very high concentrations of the organic acids reduced the amounts of Cry1Ac in the midgut of H. armigera larvae. Organic acids in the artificial diet also increased the excretion of Cry1Ac in the fecal matter. Organic acids reduced the amount of protein in the BBMV of insects reared on diets with Cry1Ac, possibly because of reduced size of the larvae. Oxalic and malic acids at concentrations present in chickpea leaves did not affect the biological activity of Cry1Ac, but it will be desirable to have high levels of expression of Cry1Ac toxin proteins in chickpea for effective control of the pod borer, H. armigera. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. A bifunctional α-amylase/trypsin inhibitor from pigeonpea seeds: Purification, biochemical characterization and its bio-efficacy against Helicoverpa armigera.

    PubMed

    Gadge, Prafull P; Wagh, Sandip K; Shaikh, Faiyaz K; Tak, Rajesh D; Padul, Manohar V; Kachole, Manvendra S

    2015-11-01

    This paper evaluates α-amylase inhibitor (α-AI) mediated defense of pigeonpea against Helicoverpa armigera. A bifunctional α-amylase/trypsin inhibitor was purified from the seeds of pigeonpea by native liquid phase isoelectric focusing (N-LP-IEF), affinity chromatography and preparative electrophoresis. Its in-vivo and in-vitro interaction with midgut amylases of H. armigera was studied along with growth inhibitory activity. One and two dimensional (2D) zymographic analyses revealed that the purified inhibitor is dimeric glycoprotein (60.2kDa and 56kDa) exist in a multi-isomeric form with five pI variants (pI 5.5 to 6.3). It was found to be heat labile with complete inactivation up to 80°C and stable over a wide range of pH (4-11). The slow binding and competitive type of α-amylase inhibition was observed with 0.08μM of dissociation constant (Ki) for the enzyme-inhibitor complex (EI). The internal protein sequence of two subunits obtained by mass spectrometry matched with cereal-type α-AI, a conserved domain from AAI_LTSS superfamily and sialyltransferase-like protein respectively. In-vivo studies indicated up-regulation of total midgut α-amylase activity with negative effect on growth rate of H. armigera suggesting its suitability for pest control. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Identification and expression patterns of Halloween genes encoding cytochrome P450s involved in ecdysteroid biosynthesis in the cotton bollworm Helicoverpa armigera.

    PubMed

    Zheng, J; Tian, K; Yuan, Y; Li, M; Qiu, X

    2017-02-01

    20-Hydroxyecdysone (20E) is a key hormone which regulates growth, development and reproduction in insects. Although cytochrome P450 enzymes (P450s) participating in the ecdysteroid biosynthesis of 20E have been characterized in a few model insects, no work has been published on the molecular entity of their orthologs in the cotton bollworm Helicoverpa armigera, a major pest insect in agriculture worldwide. In this study, four cytochrome P450 homologs, namely HarmCYP302A1, HarmCYP306A1, HarmCYP314A1 and HarmCYP315A1 from H. armigera, were identified and evolutional conservation of these Halloween genes were revealed among lepidopteran. Expression analyses showed that HarmCYP302A1 and HarmCYP315A1 were predominantly expressed in larval prothoracic glands, whereas this predominance was not always observed for HarmCYP306A1 and CYP314A1. The expression patterns of Halloween genes indicate that the fat bodies may play an important role in the conversion of ecdysone into 20E in larval-larval molt and in larval-pupal metamorphosis, and raise the possibility that HarmCYP315A1 plays a role in tissue-specific regulation in the steroid biosynthesis in H. armigera. These findings represent the first identification and expression characterization of four steriodogenic P450 genes and provide the groundwork for future functional and evolutionary study of steroid biosynthesis in this agriculturally important pest.

  20. Novel and environmental friendly approach; Impact of Neem (Azadirachta indica) gum nano formulation (NGNF) on Helicoverpa armigera (Hub.) and Spodoptera litura (Fab.).

    PubMed

    Kamaraj, Chinnaperumal; Gandhi, Pachiyappan Rajiv; Elango, Gandhi; Karthi, Sengodan; Chung, Ill-Min; Rajakumar, Govindasamy

    2017-08-30

    The future of this study was to prepare a natural pesticide which will not harm the environment and yet control pests. Neem gum nano formulation (NGNF), a novel biopesticide prepared from the Neem gum extract (Azadirachta indica) (NGE) was evaluated for its antifeedant, larvicidal and pupicidal activities against Helicoverpa armigera (Hub.) and Spodoptera litura (Fab.) at 100ppm. The NGNF showed significant (100%) antifeedant, larvicidal and pupicidal activities against H. armigera and S. litura. The LC50 values of 10.20, 12.49 and LC90 values of 32.68, 36.68ppm on H. armigera and S. litura, respectively at 100ppm. The NGNF treatments showed differences in the activities of detoxifying enzymes, carboxylesterases, glucosidases and glutathione S-transferases in the larval gut. Earthworm toxicity illustrated that 6.25ppm of chemical insecticides (cypermethrin) varied widely in their contact toxicities compared to 100ppm of NGNF and control in both contact filter paper and artificial soil test. The NGNF were characterized and confirmed by FTIR, XRD, SEM and EDX analysis. Ten compounds were identified from the Neem gum extract (NGE) by Gas Chromatography-Mass Spectrometry (GC-MS) analysis. The major compounds were fatty acids like Hexadecanoic acid, oleic acid, and ricinoleic acid. NGNF could be used as an agent to prepare novel bio-pesticides formulations. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Recombinantly expressed isoenzymic aminopeptidases from Helicoverpa armigera (American cotton bollworm) midgut display differential interaction with closely related Bacillus thuringiensis insecticidal proteins.

    PubMed Central

    Rajagopal, R; Agrawal, Neema; Selvapandiyan, Angamuthu; Sivakumar, S; Ahmad, Suhail; Bhatnagar, Raj K

    2003-01-01

    Several investigators have independently identified membrane-associated aminopeptidases in the midgut of insect larvae as the initial interacting ligand to the insecticidal crystal proteins of Bacillus thuringiensis. Though several isoenzymes of aminopeptidases have been identified from the midgut of an insect and their corresponding cDNA cloned, only one of the isoform has been expressed heterologously and studied for its binding to Cry toxins. Here we report the cloning and expression of two aminopeptidases N from Helicoverpa armigera (American cotton bollworm) (HaAPNs). The full-length cDNA of H. armigera APN1 (haapn1) is 3205 bp in size and encodes a 1000-amino-acid protein, while H. armigera APN2 (haapn2) is 3116 bp in size and corresponds to a 1012-amino-acid protein. Structurally these proteins show sequence similarity to other insect aminopeptidases and possess characteristic aminopeptidase motifs. Both the genes have been expressed in Trichoplusia ni (cabbage looper) cells using a baculovirus expression vector. The expressed aminopeptidases are membrane-associated, catalytically active and glycosylated. Ligand-blot analysis of both these aminopeptidases with bioactive Cry1Aa, Cry1Ab and Cry1Ac proteins displayed differential interaction. All the three toxins bound to HaAPN1, whereas only Cry1Ac interacted with HaAPN2. This is the first report demonstrating differential Cry-toxin-binding abilities of two different aminopeptidases from a susceptible insect. PMID:12441000

  2. Organophosphorus insecticides synergize pyrethroids in the resistant strain of cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) from West Africa.

    PubMed

    Martin, Thibaud; Ochou, Ochou G; Vaissayre, Maurice; Fournier, Didier

    2003-04-01

    Helicoverpa armigera (Hübner) populations from West Africa recently developed resistance to pyrethroid insecticides through enhanced metabolism by mixed-function oxidases. The combination index method was used to study the synergism of pyrethroids by organophosphorus insecticides. Several mixtures of insecticides currently registered to control cotton pest complex in West Africa were tested, including: cypermethrin/ethion, cypermethrin/profenofos, deltamethrin/ triazophos, deltamethrin/chlorpyriphos, cyfluthrin/chlorpyriphos, and betacyfluthrin/chlorpyriphos. In the resistant strain, the organophosphorus insecticides significantly increased the toxicity of pyrethroids suppressing the resistance effect, either by additive or synergistic effects. Significant synergism was shown for the following mixtures: cypermethrin/ethion, deltamethrin/triazophos, and deltamethrin/chlorpyriphos. The use of synergism from these insecticide mixtures should prove to be an additional tool in the overall resistance management strategy because the pyrethroid resistance in H. armigera from West Africa is not yet stable, decreasing between cotton seasons and increasing with treatments. In absence of selection, the susceptibility of H. armigera to insecticides should be restored.

  3. Effects of soil temperature and snow cover on the mortality of overwintering pupae of the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

    NASA Astrophysics Data System (ADS)

    Huang, Jian

    2016-07-01

    Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is one of the most damaging insect pests in the world. However, little is known about the effects of snow cover and soil temperature on the overwintering pupae of H. armigera. A field experiment was conducted from November 2, 2012 to April 24, 2013 at the agrometeorological experimental station in Wulanwusu, China. Overwintering pupae were embedded into the soil at depths of 5, 10, and 15 cm in the following four treatments: without snow cover, snow cover, and increased temperatures from 600 and 1200 W infrared lights. The results showed that snow cover and rising temperatures could all markedly increase soil temperatures, which was helpful in improving the survival of the overwintering pupae of H. armigera. The mortality of overwintering pupae (MOP) at a depth of 15 cm was the highest, and the MOP at a depth of 5 cm followed. The lower accumulated temperature (≤0 °C) (AT ≤ °C) led to the higher MOP, and the lower diurnal soil temperature range (DSTR) likely led to the lower MOP. After snowmelt, the MOPs at the depths of 5 and 10 cm increased as the soil temperature increased, especially in April. The AT of the soil (≤0 °C) was the factor with the strongest effect on MOP. The soil moisture content was not a major factor affecting the MOP in this semiarid region because precipitation was 45 mm over the entire experimental period. With climate warming, the MOP will likely decrease, and the overwintering boundary air temperatures of H. armigera should be expanded due to higher soil temperatures and increased snow cover.

  4. Purification and Partial Characterization of Trypsin-Specific Proteinase Inhibitors from Pigeonpea Wild Relative Cajanus platycarpus L. (Fabaceae) Active against Gut Proteases of Lepidopteran Pest Helicoverpa armigera

    PubMed Central

    Swathi, Marri; Mishra, Prashant K.; Lokya, Vadthya; Swaroop, Vanka; Mallikarjuna, Nalini; Dutta-Gupta, Aparna; Padmasree, Kollipara

    2016-01-01

    Proteinase inhibitors (PIs) are natural defense proteins of plants found to be active against gut proteases of various insects. A pigeonpea wild relative Cajanus platycarpus was identified as a source of resistance against Helicoverpa armigera, a most devastating pest of several crops including pigeonpea. In the light of earlier studies, trypsin-specific PIs (CpPI 63) were purified from mature dry seeds of C. platycarpus (ICPW-63) and characterized their biochemical properties in contributing to H. armigera resistance. CpPI 63 possessed significant H. armigera gut trypsin-like proteinase inhibitor (HGPI) activity than trypsin inhibitor (TI) activity. Analysis of CpPI 63 using two-dimensional (2-D) electrophoresis and matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that it contained several isoinhibitors and small oligomers with masses ranging between 6 and 58 kDa. The gelatin activity staining studies suggest that these isoinhibitors and oligomers possessed strong inhibitory activity against H. armigera gut trypsin-like proteases (HGPs). The N-terminal sequence of the isoinhibitors (pI 6.6 and pI 5.6) of CpPI 63 exhibited 80% homology with several Kunitz trypsin inhibitors (KTIs) as well as miraculin-like proteins (MLPs). Further, modification of lysine residue(s) lead to 80% loss in both TI and HGPI activities of CpPI 63. In contrast, the TI and HGPI activities of CpPI 63 were stable over a wide range of temperature and pH conditions. The reported results provide a biochemical basis for pod borer resistance in C. platycarpus. PMID:27656149

  5. Effects of soil temperature and snow cover on the mortality of overwintering pupae of the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

    PubMed

    Huang, Jian

    2016-07-01

    Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is one of the most damaging insect pests in the world. However, little is known about the effects of snow cover and soil temperature on the overwintering pupae of H. armigera. A field experiment was conducted from November 2, 2012 to April 24, 2013 at the agrometeorological experimental station in Wulanwusu, China. Overwintering pupae were embedded into the soil at depths of 5, 10, and 15 cm in the following four treatments: without snow cover, snow cover, and increased temperatures from 600 and 1200 W infrared lights. The results showed that snow cover and rising temperatures could all markedly increase soil temperatures, which was helpful in improving the survival of the overwintering pupae of H. armigera. The mortality of overwintering pupae (MOP) at a depth of 15 cm was the highest, and the MOP at a depth of 5 cm followed. The lower accumulated temperature (≤0 °C) (AT ≤ °C) led to the higher MOP, and the lower diurnal soil temperature range (DSTR) likely led to the lower MOP. After snowmelt, the MOPs at the depths of 5 and 10 cm increased as the soil temperature increased, especially in April. The AT of the soil (≤0 °C) was the factor with the strongest effect on MOP. The soil moisture content was not a major factor affecting the MOP in this semiarid region because precipitation was 45 mm over the entire experimental period. With climate warming, the MOP will likely decrease, and the overwintering boundary air temperatures of H. armigera should be expanded due to higher soil temperatures and increased snow cover.

  6. Oviposition site selection and survival of susceptible and resistant larvae of Helicoverpa armigera (Lepidoptera: Noctuidae) on Bt and non-Bt cotton.

    PubMed

    Luong, T T A; Downes, S J; Cribb, B; Perkins, L E; Zalucki, M P

    2016-12-01

    In Australia Bt cotton has been planted since 1996, and has greatly improved the control of its key target Helicoverpa armigera (Hübner). There is no strong evidence that genetically modified cotton has been selected for significant physiological resistance to Bt toxin in field populations. There are many possible explanations for the lack of apparent selection that range from high compliance with the resistance management strategy for this technology to a lack of behavioral preference in key traits such as oviposition that could favor survival. To date most experiments that test oviposition of H. armigera on Bt cotton vs. conventional cotton have been done with susceptible moths. We determine the oviposition preference of a field isolated Bt resistant line of H. armigera and a susceptible counterpart when given a choice of non-Bt cotton and Bt-cotton with the same genetic background, and test whether there is any relationship between oviposition site selection (different plant structures) and the survival of the first instar larvae. Within cotton plants, our experiments consistently showed that both resistant and susceptible moths did not choose plants or plant parts that were less toxic in terms of Bt toxin on which to lay eggs. There was one exception in that susceptible moths were more likely to lay eggs on squares of Bt cotton plants than squares of non-Bt cotton. As expected, the mortality of susceptible H. armigera neonates was significantly higher on structures of Bt cotton plants than on those structures of conventional cotton, and survival was greater on flowers than on other structures of Bt cotton. This confirms opportunities for selection for resistance, and demonstrates no advantage in this respect to carrying resistance genes that might overcome the Bt toxins.

  7. Population Genetic Structure of the Cotton Bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in India as Inferred from EPIC-PCR DNA Markers

    PubMed Central

    Behere, Gajanan Tryambak; Tay, Wee Tek; Russell, Derek Alan; Kranthi, Keshav Raj; Batterham, Philip

    2013-01-01

    Helicoverpa armigera is an important pest of cotton and other agricultural crops in the Old World. Its wide host range, high mobility and fecundity, and the ability to adapt and develop resistance against all common groups of insecticides used for its management have exacerbated its pest status. An understanding of the population genetic structure in H. armigera under Indian agricultural conditions will help ascertain gene flow patterns across different agricultural zones. This study inferred the population genetic structure of Indian H. armigera using five Exon-Primed Intron-Crossing (EPIC)-PCR markers. Nested alternative EPIC markers detected moderate null allele frequencies (4.3% to 9.4%) in loci used to infer population genetic structure but the apparently genome-wide heterozygote deficit suggests in-breeding or a Wahlund effect rather than a null allele effect. Population genetic analysis of the 26 populations suggested significant genetic differentiation within India but especially in cotton-feeding populations in the 2006–07 cropping season. In contrast, overall pair-wise FST estimates from populations feeding on food crops indicated no significant population substructure irrespective of cropping seasons. A Baysian cluster analysis was used to assign the genetic make-up of individuals to likely membership of population clusters. Some evidence was found for four major clusters with individuals in two populations from cotton in one year (from two populations in northern India) showing especially high homogeneity. Taken as a whole, this study found evidence of population substructure at host crop, temporal and spatial levels in Indian H. armigera, without, however, a clear biological rationale for these structures being evident. PMID:23326431

  8. Population genetic structure of the cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in India as inferred from EPIC-PCR DNA markers.

    PubMed

    Behere, Gajanan Tryambak; Tay, Wee Tek; Russell, Derek Alan; Kranthi, Keshav Raj; Batterham, Philip

    2013-01-01

    Helicoverpa armigera is an important pest of cotton and other agricultural crops in the Old World. Its wide host range, high mobility and fecundity, and the ability to adapt and develop resistance against all common groups of insecticides used for its management have exacerbated its pest status. An understanding of the population genetic structure in H. armigera under Indian agricultural conditions will help ascertain gene flow patterns across different agricultural zones. This study inferred the population genetic structure of Indian H. armigera using five Exon-Primed Intron-Crossing (EPIC)-PCR markers. Nested alternative EPIC markers detected moderate null allele frequencies (4.3% to 9.4%) in loci used to infer population genetic structure but the apparently genome-wide heterozygote deficit suggests in-breeding or a Wahlund effect rather than a null allele effect. Population genetic analysis of the 26 populations suggested significant genetic differentiation within India but especially in cotton-feeding populations in the 2006-07 cropping season. In contrast, overall pair-wise F(ST) estimates from populations feeding on food crops indicated no significant population substructure irrespective of cropping seasons. A Baysian cluster analysis was used to assign the genetic make-up of individuals to likely membership of population clusters. Some evidence was found for four major clusters with individuals in two populations from cotton in one year (from two populations in northern India) showing especially high homogeneity. Taken as a whole, this study found evidence of population substructure at host crop, temporal and spatial levels in Indian H. armigera, without, however, a clear biological rationale for these structures being evident.

  9. Artificial miRNA-mediated silencing of ecdysone receptor (EcR) affects larval development and oogenesis in Helicoverpa armigera.

    PubMed

    Yogindran, Sneha; Rajam, Manchikatla Venkat

    2016-10-01

    The insect pests are real threat to farmers as they affect the crop yield to a great extent. The use of chemical pesticides for insect pest control has always been a matter of concern as they pollute the environment and are also harmful for human health. Bt (Bacillus thuringensis) technology helped the farmers to get rid of the insect pests, but experienced a major drawback due to the evolution of insects gaining resistance towards these toxins. Hence, alternative strategies are high on demand to control insect pests. RNA-based gene silencing is emerging as a potential tool to tackle with this problem. In this study, we have shown the use of artificial microRNA (amiRNA) to specifically target the ecdysone receptor (EcR) gene of Helicoverpa armigera (cotton bollworm), which attacks several important crops like cotton, tomato chickpea, pigeon pea, etc and causes huge yield losses. Insect let-7a precursor miRNA (pre-miRNA) backbone was used to replace the native miRNA with that of amiRNA. The precursor backbone carrying the 21 nucleotide amiRNA sequence targeting HaEcR was cloned in bacterial L4440 vector for in vitro insect feeding experiments. Larvae fed with Escherichia coli expressing amiRNA-HaEcR showed a reduction in the expression of target gene as well as genes involved in the ecdysone signaling pathway downstream to EcR and exhibited mortality and developmental defects. Stem-loop RT-PCR revealed the presence of amiRNA in the insect larvae after feeding bacteria expressing amiRNA-HaEcR, which was otherwise absent in controls. We also found a significant drop in the reproduction potential (oogenesis) of moths which emerged from treated larvae as compared to control. These results demonstrate the successful use of an insect pre-miRNA backbone to express amiRNA for gene silencing studies in insects. The method is cost effective and can be exploited as an efficient and alternative tool for insect pest management.

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

    PubMed

    Cai, Jun; Xiao, Liang; Yan, Bing; Bin, Guan; Chen, Yuehua; Ren, Gaixin

    2006-04-01

    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.

  11. Negative Effects of a Nonhost Proteinase Inhibitor of ~19.8 kDa from Madhuca indica Seeds on Developmental Physiology of Helicoverpa armigera (Hübner)

    PubMed Central

    Jamal, Farrukh; Singh, Dushyant; Pandey, Prabhash K.

    2014-01-01

    An affinity purified trypsin inhibitor from the seed flour extracts of Madhuca indica (MiTI) on denaturing polyacrylamide gel electrophoresis showed that MiTI consisted of a single polypeptide chain with molecular mass of ~19.8 kDa. MiTI inhibited the total proteolytic and trypsin-like activities of the midgut proteinases of Helicoverpa armigera larvae by 87.51% and 76.12%, respectively, at concentration of 5 µg/mL with an IC50 of 1.75 µg/mL against trypsin like midgut proteinases. The enzyme kinetic studies demonstrated that MiTI is a competitive inhibitor with a Ki value of 4.1 × 10−10 M for Helicoverpa trypsin like midgut proteinases. In vivo experiments with different concentrations of MiTI in artificial diet (0.5, 1.0, and 1.5% w/w) showed an effective downfall in the larval body weight and an increase in larval mortality. The concentration of MiTI in the artificial diet to cause 50% mortality (LD50) of larvae was 1.5% w/w and that to cause reduction in mass of larvae by 50% (ED50) was 1.0% w/w. Nutritional indices observations suggest the toxic and adverse effects of MiTI on the growth and development of H. armigera larvae. The results suggest a strong bioinsecticidal potential of affinity purified MiTI which can be exploited in insect pest management of crop plants. PMID:25298962

  12. Effects of Soil Salinity on the Expression of Bt Toxin (Cry1Ac) and the Control Efficiency of Helicoverpa armigera in Field-Grown Transgenic Bt Cotton.

    PubMed

    Luo, Jun-Yu; Zhang, Shuai; Peng, Jun; Zhu, Xiang-Zhen; Lv, Li-Min; Wang, Chun-Yi; Li, Chun-Hua; Zhou, Zhi-Guo; Cui, Jin-Jie

    2017-01-01

    An increasing area of transgenic Bacillus thuringiensis (Bt) cotton is being planted in saline-alkaline soil in China. The Bt protein level in transgenic cotton plants and its control efficiency can be affected by abiotic stress, including high temperature, water deficiency and other factors. However, how soil salinity affects the expression of Bt protein, thus influencing the control efficiency of Bt cotton against the cotton bollworm (CBW) Helicoverpa armigera (Hübner) in the field, is poorly understood. Our objective in the present study was to investigate the effects of soil salinity on the expression of Bt toxin (Cry1Ac) and the control efficiency of Helicoverpa armigera in field-grown transgenic Bt cotton using three natural saline levels (1.15 dS m-1 [low soil-salinity], 6.00 dS m-1 [medium soil-salinity] and 11.46 dS m-1 [high soil-salinity]). We found that the Bt protein content in the transgenic Bt cotton leaves and the insecticidal activity of Bt cotton against CBW decreased with the increasing soil salinity in laboratory experiments during the growing season. The Bt protein content of Bt cotton leaves in the laboratory were negatively correlated with the salinity level. The CBW populations were highest on the Bt cotton grown in medium-salinity soil instead of the high-salinity soil in field conditions. A possible mechanism may be that the relatively high-salinity soil changed the plant nutritional quality or other plant defensive traits. The results from this study may help to identify more appropriate practices to control CBW in Bt cotton fields with different soil salinity levels.

  13. Effects of Soil Salinity on the Expression of Bt Toxin (Cry1Ac) and the Control Efficiency of Helicoverpa armigera in Field-Grown Transgenic Bt Cotton

    PubMed Central

    Luo, Jun-Yu; Zhang, Shuai; Peng, Jun; Zhu, Xiang-Zhen; Lv, Li-Min; Wang, Chun-Yi; Li, Chun-Hua; Zhou, Zhi-Guo; Cui, Jin-Jie

    2017-01-01

    An increasing area of transgenic Bacillus thuringiensis (Bt) cotton is being planted in saline-alkaline soil in China. The Bt protein level in transgenic cotton plants and its control efficiency can be affected by abiotic stress, including high temperature, water deficiency and other factors. However, how soil salinity affects the expression of Bt protein, thus influencing the control efficiency of Bt cotton against the cotton bollworm (CBW) Helicoverpa armigera (Hübner) in the field, is poorly understood. Our objective in the present study was to investigate the effects of soil salinity on the expression of Bt toxin (Cry1Ac) and the control efficiency of Helicoverpa armigera in field-grown transgenic Bt cotton using three natural saline levels (1.15 dS m-1 [low soil-salinity], 6.00 dS m-1 [medium soil-salinity] and 11.46 dS m-1 [high soil-salinity]). We found that the Bt protein content in the transgenic Bt cotton leaves and the insecticidal activity of Bt cotton against CBW decreased with the increasing soil salinity in laboratory experiments during the growing season. The Bt protein content of Bt cotton leaves in the laboratory were negatively correlated with the salinity level. The CBW populations were highest on the Bt cotton grown in medium-salinity soil instead of the high-salinity soil in field conditions. A possible mechanism may be that the relatively high-salinity soil changed the plant nutritional quality or other plant defensive traits. The results from this study may help to identify more appropriate practices to control CBW in Bt cotton fields with different soil salinity levels. PMID:28099508

  14. Effects of dietary sodium on performance, flight and compensation strategies in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

    PubMed Central

    2010-01-01

    Background Sodium is critical for many physiological functions in insects. Herbivorous insects should expend considerable energy to compensate for sodium deficiency due to low sodium concentration in most inland plants upon which they feed. However, sodium compensation behaviors such as mud-puddling have been observed in some species but not in others. We expect that there may be other sodium compensation strategies in insects. Here, we select a rarely mud-puddling insect species, the cotton boll worm, Helicoverpa armigera, and determine the effects of dietary sodium on performance and flight, and examine their means of sodium compensation. Results When freshly hatched H. armigera neonates were cultured on one of three diets differing in sodium contents (diet A, B and C with a high, middle and low sodium concentrations, respectively), the larvae on diet C grew larger, had a higher mortality rate and a shorter development period than those on diet A and B. The larvae previously fed from 1st to 3rd instar on diet C consumed more subsequent diet when they were transferred to diet A or C at 4th instar, comparing to those previously fed on diet A. Moreover, any 4th-instar larvae on diet C consumed a greater amount of food than those on diet A, no matter which diet the larvae had previously ingested from 1st to 3rd instar. Moths from diet A and B flew more rapidly than those from diet C, with similar sugar and lipid utilization rates among the three test groups. When a 5th-instar cannibal from diet A, B or C and a 5th-instar victim from diet A were housed together, many more cannibals from diet C ate their victims. When a victim from diet A, B or C was provided, a cannibal from diet C was more likely to eat the victim from diet A. When newly emerged moths had been exposed to 3% sodium chloride solution for all scotophase period, the average weight increase (proxy for sodium solution intake) for moths from diet A was lower than those from diet B or C. Conclusion Sodium

  15. Effects of dietary sodium on performance, flight and compensation strategies in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

    PubMed

    Xiao, Kai; Shen, Ke; Zhong, Jian-Feng; Li, Guo-Qing

    2010-04-13

    Sodium is critical for many physiological functions in insects. Herbivorous insects should expend considerable energy to compensate for sodium deficiency due to low sodium concentration in most inland plants upon which they feed. However, sodium compensation behaviors such as mud-puddling have been observed in some species but not in others. We expect that there may be other sodium compensation strategies in insects. Here, we select a rarely mud-puddling insect species, the cotton boll worm, Helicoverpa armigera, and determine the effects of dietary sodium on performance and flight, and examine their means of sodium compensation. When freshly hatched H. armigera neonates were cultured on one of three diets differing in sodium contents (diet A, B and C with a high, middle and low sodium concentrations, respectively), the larvae on diet C grew larger, had a higher mortality rate and a shorter development period than those on diet A and B. The larvae previously fed from 1st to 3rd instar on diet C consumed more subsequent diet when they were transferred to diet A or C at 4th instar, comparing to those previously fed on diet A. Moreover, any 4th-instar larvae on diet C consumed a greater amount of food than those on diet A, no matter which diet the larvae had previously ingested from 1st to 3rd instar. Moths from diet A and B flew more rapidly than those from diet C, with similar sugar and lipid utilization rates among the three test groups. When a 5th-instar cannibal from diet A, B or C and a 5th-instar victim from diet A were housed together, many more cannibals from diet C ate their victims. When a victim from diet A, B or C was provided, a cannibal from diet C was more likely to eat the victim from diet A. When newly emerged moths had been exposed to 3% sodium chloride solution for all scotophase period, the average weight increase (proxy for sodium solution intake) for moths from diet A was lower than those from diet B or C. Sodium-deficient diet resulted in rapid

  16. Frequency of alleles conferring resistance to the Bt toxins Cry1Ac and Cry2Ab in Australian populations of Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Mahon, R J; Olsen, K M; Downes, S; Addison, S

    2007-12-01

    Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is an important lepidopteran pest of cotton (Gossypium spp.) in Australia and the Old World. From 2002, F2 screens were used to examine the frequency of resistance alleles in Australian populations of H. armigera to Bacillus thuringiensis (Bt) CrylAc and Cry2Ab, the two insecticidal proteins present in the transgenic cotton Bollgard II. At that time, Ingard (expressing Cry1Ac) cotton had been grown in Australia for seven seasons, and Bollgard II was about to be commercially released. The principal objective of our study was to determine whether sustained exposure caused an elevated frequency of alleles conferring resistance to Cry1Ac in a species with a track record of evolving resistance to conventional insecticides. No major alleles conferring resistance to Cry1Ac were found. The frequency of resistance alleles for Cry1Ac was <0.0003, with a 95% credibility interval between 0 and 0.0009. In contrast, alleles conferring resistance to Cry2Ab were found at a frequency of 0.0033 (0.0017, 0.0055). The first isolation of this allele was found before the widespread deployment of Bollgard II. For both toxins the experiment-wise detection probability was 94.4%. Our results suggest that alleles conferring resistance to Cry1Ac are rare and that a relatively high baseline frequency of alleles conferring resistance to Cry2Ab existed before the introduction of Bt cotton containing this toxin.

  17. Transgenic plants over-expressing insect-specific microRNA acquire insecticidal activity against Helicoverpa armigera: an alternative to Bt-toxin technology.

    PubMed

    Agrawal, Aditi; Rajamani, Vijayalakshmi; Reddy, Vanga Siva; Mukherjee, Sunil Kumar; Bhatnagar, Raj K

    2015-10-01

    The success of Bt transgenics in controlling predation of crops has been tempered by sporadic emergence of resistance in targeted insect larvae. Such emerging threats have prompted the search for novel insecticidal molecules that are specific and could be expressed through plants. We have resorted to small RNA-based technology for an investigative search and focused our attention to an insect-specific miRNA that interferes with the insect molting process resulting in the death of the larvae. In this study, we report the designing of a vector that produces artificial microRNA (amiR), namely amiR-24, which targets the chitinase gene of Helicoverpa armigera. This vector was used as transgene in tobacco. Northern blot and real-time analysis revealed the high level expression of amiR-24 in transgenic tobacco plants. Larvae feeding on the transgenic plants ceased to molt further and eventually died. Our results demonstrate that transgenic tobacco plants can express amiR-24 insectice specific to H. armigera.

  18. Kinetic assessment and effect on developmental physiology of a trypsin inhibitor from Eugenia jambolana (Jambul) seeds on Helicoverpa armigera (Hübner).

    PubMed

    Singh, Dushyant; Jamal, Farrukh; Pandey, Prabhash K

    2014-02-01

    A trypsin inhibitor was purified from the seeds of Eugenia jambolana (Jambul) with a fold purification of 14.28 and a yield recovery of 2.8%. Electrophoretic analysis of E. jambolana trypsin inhibitor (EjTI) revealed a molecular weight of approximately 17.4 kDa on 12% denaturing polyacrylamide gel electrophoresis with or without reduction. EjTI exhibited high stability over a wide range of temperatures (4-80 °C for 30 min) and pH (3.0-10.0) and inhibited trypsin-like activities of the midgut proteinases of fourth instar Helicoverpa armigera larvae by approximately 86%. Feeding assays containing 0.05, 0.15, and 0.45 (% w/w) EjTI on functionally important fourth-instar larvae indicated a dose-dependent downfall in the larval body weight as well as on extent of survival. The nutritional analysis suggests that EjTI exerts toxic effects on H. armigera. Dixon plot analysis revealed competitive inhibition of larval midgut proteinases by EjTI, with an inhibition constant (Ki ) of approximately 3.1 × 10(-9) M. However, inhibitor kinetics using double reciprocal plots for trypsin inhibition demonstrated a mixed inhibition pattern. These observations suggest the potential of E. jambolana trypsin inhibitor protein in insect pest management. © 2014 Wiley Periodicals, Inc.

  19. Isomer-specific comparisons of the hydrolysis of synthetic pyrethroids and their fluorogenic analogues by esterases from the cotton bollworm Helicoverpa armigera.

    PubMed

    Yuan, G; Li, Y; Farnsworth, C A; Coppin, C W; Devonshire, A L; Scott, C; Russell, R J; Wu, Y; Oakeshott, J G

    2015-06-01

    The low aqueous solubility and chiral complexity of synthetic pyrethroids, together with large differences between isomers in their insecticidal potency, have hindered the development of meaningful assays of their metabolism and metabolic resistance to them. To overcome these problems, Shan and Hammock (2001) [7] therefore developed fluorogenic and more water-soluble analogues of all the individual isomers of the commonly used Type 2 pyrethroids, cypermethrin and fenvalerate. The analogues have now been used in several studies of esterase-based metabolism and metabolic resistance. Here we test the validity of these analogues by quantitatively comparing their hydrolysis by a battery of 22 heterologously expressed insect esterases with the hydrolysis of the corresponding pyrethroid isomers by these esterases in an HPLC assay recently developed by Teese et al. (2013) [14]. We find a strong, albeit not complete, correlation (r = 0.7) between rates for the two sets of substrates. The three most potent isomers tested were all relatively slowly degraded in both sets of data but three esterases previously associated with pyrethroid resistance in Helicoverpa armigera did not show higher activities for these isomers than did allelic enzymes derived from susceptible H. armigera. Given their amenability to continuous assays at low substrate concentrations in microplate format, and ready detection of product, we endorse the ongoing utility of the analogues in many metabolic studies of pyrethroids. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  20. Fitness costs of reproductive capacity and ovarian development in a Bt-resistant strain of the cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

    PubMed

    Zhang, Wanna; Ma, Long; Zhong, Feng; Wang, Yanan; Guo, Yuyuan; Lu, Yanhui; Liang, Gemei

    2015-06-01

    The cotton bollworm Helicoverpa armigera has developed resistance to the insecticidal Cry1Ac toxins produced by Bacillus thuringiensis (Bt) in China. To gain a better understanding of the fitness cost associated with resistance evolution, reproductive capacities and ovarian development were examined in four strains of H. armigera: a Cry1Ac-susceptible strain (96S); a Cry1Ac-resistant strain fed on an artificial diet with Cry1Ac toxin for 135 generations (BtR); two strains derived from BtR: a strain grown without Cry1Ac selection for 38 generations (CK1) and a strain grown for one generation without Cry1Ac selection (CK2). Strains 96S and CK1 had similar reproductive capacity and Cry1Ac susceptibility. Compared with the 96S and CK1 strains, the fecundity and hatching rate of CK2 were respectively more than 30% and 50% lower. Moreover, the number of eggs laid by BtR was significantly lower than the number of eggs laid by 96S and CK1, but higher than the number of eggs laid by CK2. In accordance with the differences in reproductive capacity, ovarian development in CK2 and BtR was significantly delayed. These results indicated that there was a fitness cost in the reproductive physiology in the resistant strains. Reversion to susceptibility to the Bt Cry-toxin in the resistant CK1 strain was associated with an increase in reproductive capacity. © 2014 Society of Chemical Industry.

  1. Presence of snow coverage and its thickness affected the mortality of overwintering pupae of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

    NASA Astrophysics Data System (ADS)

    Huang, Jian

    2016-10-01

    Helicoverpa armigera causes serious damage to most crops around the world. However, the impacts of snow thickness on the H. armigera overwintering pupae are little known. A field experiment was employed in 2012-2015 at Urumqi, China. At soil depths of 5, 10, and 15 cm, overwintering pupae were embedded with four treatments: no snow cover (NSC), snow cover (SC), increasing snow thickness to 1.5 times the thickness of SC (ISSC-1), and to two times the thickness of SC (ISSC-2). Results suggested that snow cover and increasing snow thickness both significantly increased soil temperatures, which helped to decrease the mortality of overwintering pupae (MOP) of H. armigera. However, the MOP did not always decrease with increases in snow thickness. The MOPs in NSC and ISSC-1 were the highest and the lowest, respectively, though ISSC-2 had much thicker snow thickness than ISSC-1. A maximum snow thickness of 60 cm might lead to the lowest MOP. The longer the snow cover duration (SCD) at a soil depth of 10 cm in March and April was, the higher the MOP was. A thicker snow cover layer led to a higher soil moisture content (SMC) and a lower diurnal soil temperature range (DSTR). The highest and the lowest MOP were at a depth of 15 and 10 cm, respectively. The SMC at the depths of 10 and 15 cm had significant effects on MOP. A lower accumulated temperature (≤0 °C) led to a higher MOP. The DSTR in March of approximately 4.5 °C might cause the lowest MOP. The largest influence factor for the MOPs at depths of 5 and 10 cm and the combined data were the SCDs during the whole experimental period, and for the MOPs at a depth of 15 cm was the soil temperature in November.

  2. Presence of snow coverage and its thickness affected the mortality of overwintering pupae of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

    PubMed

    Huang, Jian

    2017-04-01

    Helicoverpa armigera causes serious damage to most crops around the world. However, the impacts of snow thickness on the H. armigera overwintering pupae are little known. A field experiment was employed in 2012-2015 at Urumqi, China. At soil depths of 5, 10, and 15 cm, overwintering pupae were embedded with four treatments: no snow cover (NSC), snow cover (SC), increasing snow thickness to 1.5 times the thickness of SC (ISSC-1), and to two times the thickness of SC (ISSC-2). Results suggested that snow cover and increasing snow thickness both significantly increased soil temperatures, which helped to decrease the mortality of overwintering pupae (MOP) of H. armigera. However, the MOP did not always decrease with increases in snow thickness. The MOPs in NSC and ISSC-1 were the highest and the lowest, respectively, though ISSC-2 had much thicker snow thickness than ISSC-1. A maximum snow thickness of 60 cm might lead to the lowest MOP. The longer the snow cover duration (SCD) at a soil depth of 10 cm in March and April was, the higher the MOP was. A thicker snow cover layer led to a higher soil moisture content (SMC) and a lower diurnal soil temperature range (DSTR). The highest and the lowest MOP were at a depth of 15 and 10 cm, respectively. The SMC at the depths of 10 and 15 cm had significant effects on MOP. A lower accumulated temperature (≤0 °C) led to a higher MOP. The DSTR in March of approximately 4.5 °C might cause the lowest MOP. The largest influence factor for the MOPs at depths of 5 and 10 cm and the combined data were the SCDs during the whole experimental period, and for the MOPs at a depth of 15 cm was the soil temperature in November.

  3. Identification and Evaluation of Suitable Reference Genes for Normalization of MicroRNA Expression in Helicoverpa armigera (Lepidoptera: Noctuidae) Using Quantitative Real-Time PCR.

    PubMed

    Yang, Yuhui; Li, Zhen; Cao, Jinjun; Li, Yanrong; Li, Hui; Yang, Qingpo; Zhang, Qingwen; Liu, Xiaoxia

    2017-01-01

    More and more studies have focused on microRNAs (miRNAs) expression in the pest Helicoverpa armigera (Lepidoptera: Noctuidae) recently. Quantitative real-time PCR (qRT-PCR) is being widely used in miRNA expression studies. Suitable reference genes are necessary for the correct analysis of results. In this study, 10 candidate genes of H. armigera were selected and analyzed for their expression stability under different biotic and abiotic conditions with 3 statistical methods, including geNorm, NormFinder, and Bestkeeper. Combination the best number of reference genes was calculated by geNorm. One target gene, let-7, was used to validate the selection of reference genes. The suitable candidate reference genes were shown as follows: miR-9 and U6 snRNA for developmental stages, miR-100 and U6 snRNA for larval tissues, miR-100 and miR-305 for adult tissues, miR-9 and miR-279 for parasitic treatment, miR-998 and U6 snRNA for nuclear polyhedrosis virus infection, miR-9 and U6 snRNA for insecticide treatment, miR-92a, miR-100, and miR-279 for temperature treatment, miR-92a, miR-305, and miR-998 for starvation treatment, miR-9 and miR-279 for light treatment, miR-305 and miR-998 for hormone treatment, and there was not one reference gene suitable for all samples. This study could promote future research on miRNAs expression in H. armigera with optimal reference genes under different experimental conditions. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America.

  4. Tracking pyrethroid resistance in the polyphagous bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae), in the shifting landscape of a cotton-growing area.

    PubMed

    Brévault, T; Achaleke, J; Sougnabé, S P; Vaissayre, M

    2008-12-01

    In cotton-growing areas of Central Africa, timing of host crops and pest management practices in annual rainfed cropping systems result in a shifting mosaic of habitats that influence the dynamics and resistance of Helicoverpa armigera (Hübner) populations on spatial scales, both within and across seasons. From 2002 to 2006, regional and local resistance was monitored among cotton fields and among the major host plants of the bollworm. From 2002, pyrethroid resistance increased within and across cotton-growing seasons to reach a worrying situation at the end of the 2005 growing season. Cotton crops played a fundamental role in the increase in seasonal resistance, even if the intensive use of insecticides on local tomato crops strongly concentrated resistance alleles in residual populations throughout the off-season. Due to the relative stability of resistance in H. armigera populations despite a long off-season, we believe that after the dispersal of the moths southwards at the end of the growing season, reverse migration mainly accounts for the reconstitution of populations at the onset of the following growing season. In addition, local resistance monitoring in 2005 and 2006 showed that it was possible to control the increase in resistance by temporarily stopping the use of pyrethroids during the period of peak infestation of cotton by H. armigera. On the other hand, the similar resistance frequency of populations sampled from sprayed and unsprayed synchronous hosts confirmed the absence of reproductive isolation between adults. As a result, diversity in cropping systems should be encouraged by planting alternative host plants to provide a mosaic of habitats, which in return would provide insecticide-free refuges. The implications for insecticide resistance management in annual cropping systems are discussed.

  5. Yeast one-hybrid screening the potential regulator of CYP6B6 overexpression of Helicoverpa armigera under 2-tridecanone stress.

    PubMed

    Zhao, J; Liu, X N; Li, F; Zhuang, S Z; Huang, L N; Ma, J; Gao, X W

    2016-04-01

    In insect, the cytochrome P450 plays a pivotal role in detoxification to toxic allelochemicals. Helicoverpa armigera can tolerate and survive in 2-tridecanone treatment owing to the CYP6B6 responsive expression, which is controlled by some regulatory DNA sequences and transcription regulators. Therefore, the 2-tridecanone responsive region and transcription regulators of the CYP6B6 are responsible for detoxification of cotton bollworm. In this study, we used yeast one-hybrid to screen two potential transcription regulators of the CYP6B6 from H. armigera that respond to the plant secondary toxicant 2-tridecanone, which were named Prey1 and Prey2, respectively. According to the NCBI database blast, Prey1 is the homology with FK506 binding protein (FKBP) of Manduca sexta and Bombyx mori that belongs to the FKBP-C superfamily, while Prey2 may be a homology of an unknown protein of Papilio or the fcaL24 protein homology of B. mori. The electrophoretic mobility shift assays revealed that the FKBP of prokaryotic expression could specifically bind to the active region of the CYP6B6 promoter. After the 6th instar larvae of H. armigera reared on 2-tridecanone artificial diet, we found there were similar patterns of CYP6B6 and FKBP expression of the cotton bollworm treated with 10 mg g-1 2-tridecanone for 48 h, which correlation coefficient was the highest (0.923). Thus, the FKBP is identified as a strong candidate for regulation of the CYP6B6 expression, when the cotton bollworm is treated with 2-tridecanone. This may lead us to a better understanding of transcriptional mechanism of CYP6B6 and provide very useful information for the pest control.

  6. Transgenic Tobacco Expressing the TAT-Helicokinin I-CpTI Fusion Protein Show Increased Resistance and Toxicity to Helicoverpa armigera (Lepidoptera: Noctuidae)

    PubMed Central

    Zhou, Zhou; Li, Yongli; Yuan, Chunyan; Zhang, Yongan; Qu, Liangjian

    2017-01-01

    Insect kinins were shown to have diuretic activity, inhibit weight gain, and have antifeedant activity in insects. In order to study the potential of the TAT-fusion approach to deliver diuretic peptides per os to pest insects, the HezK I peptide from Helicoverpa zea, as a representative of the kinin family, was selected. The fusion gene TAT-HezK I was designed and was used to transform tobacco plants. As a means to further improve the stability of TAT-HezK I, a fusion protein incorporating HezK I, transactivator of transcription (TAT), and the cowpea trypsin inhibitor (CpTI) was also designed. Finally, the toxicity of the different tobacco transgenic strains toward Helicoverpa armigera was compared. The results demonstrated that TAT-HezK I had high toxicity against insects via transgenic expression of the peptide in planta and intake through larval feeding. The toxicity of the fusion TAT-HezK I and CpTI was higher than the CpTI single gene in transgenic tobacco, and the fusion TAT-HezK I and CpTI further enhanced the stability and bioavailability of agents in oral administration. Our research helps in targeting new genes for improving herbivore tolerance in transgenic plant breeding. PMID:28085119

  7. Tradeoff between reproduction and resistance evolution to Bt-toxin in Helicoverpa armigera: regulated by vitellogenin gene expression.

    PubMed

    Zhang, W N; Xiao, H J; Liang, G M; Guo, Y Y; Wu, K M

    2014-08-01

    Evolution of resistance to insecticides usually has fitness tradeoffs associated with adaptation to the stress. The basic regulation mechanism of tradeoff between reproduction and resistance evolution to Bacillus thuringiensis (Bt) toxin in the cotton bollworm, Helicoverpa armigera (Ha), based on the vitellogenin (Vg) gene expression was analyzed here. The full-length cDNA of the Vg gene HaVg (JX504706) was cloned and identified. HaVg has 5704 base pairs (bp) with an open reading frame (ORF) of 5265 bp, which encoded 1756 amino acid protein with a predicted molecular mass of 197.28 kDa and a proposed isoelectric point of 8.74. Sequence alignment analysis indicated that the amino acid sequence of HaVg contained all of the conserved domains detected in the Vgs of the other insects and had a high similarity with the Vgs of the Lepidoptera insects, especially Noctuidae. The resistance level to Cry1Ac Bt toxin and relative HaVg mRNA expression levels among the following four groups: Cry1Ac-susceptible strain (96S), Cry1Ac-resistant strain fed on artificial diet with Bt toxin for 135 generations (BtR stands for the Cry1Ac Bt resistance), progeny of the Cry1Ac-resistant strain with a non-Bt-toxin artificial diet for 38 generations (CK1) and the direct descendants of the 135th-generation resistant larvae which were fed on an artificial diet without the Cry1Ac protein (CK2) were analyzed. Compared with the 96S strain, the resistance ratios of the BtR strain, the CK1 strain and the CK2 strain were 2917.15-, 2.15- and 2037.67-fold, respectively. The maximum relative HaVg mRNA expression levels of the BtR strain were approximately 50% less than that of the 96S strain, and the coming of maximum expression was delayed for approximately 4 days. The overall trend of the HaVg mRNA expression levels in the CK1 strain was similar to that in the 96S strain, and the overall trend of the HaVg mRNA expression levels in the CK2 strain was similar to that in the BtR strain. Our results

  8. Genomic basis for the pest status of two Helicoverpa species

    USDA-ARS?s Scientific Manuscript database

    Background: Helicoverpa armigera and Helicoverpa zea are major caterpillar pests of Old and New World agriculture respectively. Both, particularly H. armigera, are extremely polyphagous, and H. armigera has developed resistance to many insecticides. Here we use comparative and population genomics an...

  9. Effect of pyramiding Bt and CpTI genes on resistance of cotton to Helicoverpa armigera (Lepidoptera: Noctuidae) under laboratory and field conditions.

    PubMed

    Cui, Jinjie; Luo, Junyu; Van Der Werf, Wopke; Ma, Yan; Xia, Jingyuan

    2011-04-01

    Transgenic cotton (Cossypium hirsutum L.) varieties, adapted to China, have been bred that express two genes for resistance to insects, the CrylAc gene from Bacillus thuringiensis (Berliner) (Bt), and a trypsin inhibitor gene from cowpea (CpTI). Effectiveness of the double gene modification in conferring resistance to cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), was studied in laboratory and field experiments. In each experiment, performance of Bt+CpTI cotton was compared with Bt cotton and to a conventional nontransgenic variety. Larval survival was lower on both types of transgenic variety, compared with the conventional cotton. Survival of first-, second-, and third-stage larvae was lower on Bt+CpTI cotton than on Bt cotton. Plant structures differed in level of resistance, and these differences were similar on Bt and Bt + CpTI cotton. Likewise, seasonal trends in level of resistance in different plant structures were similar in Bt and Bt+CpTI cotton. Both types of transgenic cotton interfered with development of sixth-stage larvae to adults, and no offspring was produced by H. armigera that fed on Bt or Bt+CpTI cotton from the sixth stage onward. First-, second-, and third-stage larvae spent significantly less time feeding on transgenic cotton than on conventional cotton, and the reduction in feeding time was significantly greater on Bt+CpTI cotton than on Bt cotton. Food conversion efficiency was lower on transgenic varieties than on conventional cotton, but there was no significant difference between Bt and Bt+CpTI cotton. In 3-yr field experimentation, bollworm densities were greatly suppressed on transgenic as compared with conventional cotton, but no significant differences between Bt and Bt+CpTI cotton were found. Overall, the results from laboratory work indicate that introduction of the CpTI gene in Bt cotton raises some components of resistance in cotton against H. armigera, but enhanced control of H. armigera under field

  10. Cloning and Tissue-Specific Expression of a Chitin Deacetylase Gene from Helicoverpa armigera (Lepidoptera: Noctuidae) and Its Response to Bacillus thuringiensis

    PubMed Central

    Han, Guoying; Li, Xiumin; Zhang, Ting; Zhu, Xiaoting

    2015-01-01

    Chitin deacetylases (CDAs) convert chitin into chitosan, the N-deacetylated form of chitin, which influences the mechanical and permeability properties of structures such as the cuticle and peritrophic matrices. In this article, a new CDA encoding gene, Hacda2, was cloned by reverse transcription-polymerase chain reaction method in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), with an open reading frame of 1,611 bp. The deduced protein composed of 536 amino acid residues with a signal peptide, a chitin-binding domain, a low-density lipoprotein receptor class A domain, and a polysaccharide deacetylase-like catalytic domain. The highest expression level of Hacda2 was detected in fat body among tissues tested in the fifth-instar larvae using real-time quantitative polymerase chain reaction method. Feeding of Bacillus thuringiensis (Bt) (Bacillales: Bacillaceae) diet changed the expression level of Hacda1, Hacda2, Hacda5a, and Hacda5b significantly and differentially in the third-instar larvae. Hacda5a and Hacda5b expression were initially down-regulated and then up-regulated, whereas, the expression level of Hacda1 and Hacda2 was suppressed constantly postfeeding on Bt diet. These results suggested that HaCDAs may be involved in the response of H. armigera larvae to Bt and may be helpful to elucidate the roles of HaCDAs in the action of Bt cry toxin. The potential of HaCDAs to be used as synergists of Bt insecticidal protein needs to be further tested. PMID:26163665

  11. A Generalist Herbivore Copes with Specialized Plant Defence: the Effects of Induction and Feeding by Helicoverpa armigera (Lepidoptera: Noctuidae) Larvae on Intact Arabidopsis thaliana (Brassicales) Plants.

    PubMed

    Zalucki, M P; Zalucki, J M; Perkins, L E; Schramm, K; Vassão, D G; Gershenzon, J; Heckel, D G

    2017-06-01

    Plants of the Brassicaceae are defended from feeding by generalist insects by constitutively-expressed and herbivory-induced glucosinolates (GS). We induced Arabidopsis plants 1, 16 and 24 h prior to allowing neonate larvae of the generalist Helicoverpa armigera to feed on whole plants for 72 h. These plants were subsequently retested with another group of neonates for a further 72 h. We used wild-type A. thaliana Col-0, and mutant lines lacking indolic GS, aliphatic GS or all GS. We hypothesized that larvae would not grow well on defended plants (WT) compared to those lacking GS, and would not grow well if plants had been primed or fed on for longer, due to the expected induced GS. There was survivorship on all lines suggesting H. armigera is a suitable generalist for these experiments. Larvae performed less well on wild-type and no indolic lines than on no aliphatic and no GS lines. Larvae distributed feeding damage extensively in all lines, more so on wild type and no-indolic lines. Contrary to expectations, larvae grew better on plants that had been induced for 1 to 16 h than on un-induced plants suggesting they moved to and selected less toxic plant parts within a heterogeneously defended plant. Performance declined on all lines if plants had been induced for 24 h, or had been fed upon for a further 72 h. However, contrary to expectation, individual and total GS did not increase after these two treatments. This suggests that Arabidopsis plants induce additional (not GS) defenses after longer induction periods.

  12. Identification of the 2-tridecanone responsive region in the promoter of cytochrome P450 CYP6B6 of the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Li, F; Liu, X N; Zhu, Y; Ma, J; Liu, N; Yang, J H

    2014-12-01

    Eukaryote transcription is controlled by regulatory DNA sequences and transcription factors, so transcriptional control of gene plays a pivotal role in gene expression. In this study, we identified the region of the CYP6B6 gene promoter of Helicoverpa armigera which responds to the plant secondary toxicant 2-tridecanone. Transient transfection assay results from five of stepwise deletion fragments linked to the luciferase reporter gene revealed that the promoter activity of each CYP6B6 fragment was significantly higher than that of their basal activity after the Sf9 cells were treated with 2-tridecanone. Among all, the fragment spanning -373 to +405 bp of the CYP6B6 promoter showed an obviously 2-tridecanone inducibility (P<0.0001), which might have the 2-tridecanone responsive element based on promoter activity. Electrophoretic mobility shift assays revealed that the nuclear protein extracted from midgut of the 6th instar larva of H. armigera, reared on 10 mg 2-tridecanone per gram artificial diet for 48 h, could specifically bind to the active region from -373 to 21 bp of the CYP6B6 promoter. The combination feature also appeared when using a shorter fragment from -292 to -154 bp of the CYP6B6 promoter. Taken together, we found a 2-tridecanone core responsive region between -292 and -154 bp of the CYP6B6 promoter. This may lead us to a better understanding of transcriptional mechanism of P450 gene and provide very useful information for the pest control.

  13. Cloning and Tissue-Specific Expression of a Chitin Deacetylase Gene from Helicoverpa armigera (Lepidoptera: Noctuidae) and Its Response to Bacillus thuringiensis.

    PubMed

    Han, Guoying; Li, Xiumin; Zhang, Ting; Zhu, Xiaoting; Li, Jigang

    2015-01-01

    Chitin deacetylases (CDAs) convert chitin into chitosan, the N-deacetylated form of chitin, which influences the mechanical and permeability properties of structures such as the cuticle and peritrophic matrices. In this article, a new CDA encoding gene, Hacda2, was cloned by reverse transcription-polymerase chain reaction method in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), with an open reading frame of 1,611 bp. The deduced protein composed of 536 amino acid residues with a signal peptide, a chitin-binding domain, a low-density lipoprotein receptor class A domain, and a polysaccharide deacetylase-like catalytic domain. The highest expression level of Hacda2 was detected in fat body among tissues tested in the fifth-instar larvae using real-time quantitative polymerase chain reaction method. Feeding of Bacillus thuringiensis (Bt) (Bacillales: Bacillaceae) diet changed the expression level of Hacda1, Hacda2, Hacda5a, and Hacda5b significantly and differentially in the third-instar larvae. Hacda5a and Hacda5b expression were initially down-regulated and then up-regulated, whereas, the expression level of Hacda1 and Hacda2 was suppressed constantly postfeeding on Bt diet. These results suggested that HaCDAs may be involved in the response of H. armigera larvae to Bt and may be helpful to elucidate the roles of HaCDAs in the action of Bt cry toxin. The potential of HaCDAs to be used as synergists of Bt insecticidal protein needs to be further tested.

  14. Identification and characterization of the Sudanese Bacillus thuringiensis and related bacterial strains for their efficacy against Helicoverpa armigera and Tribolium castaneum.

    PubMed

    Gorashi, N E; Tripathi, M; Kalia, V; Gujar, G T

    2014-06-01

    Forty-four isolates of Bacillus thuringiensis like bacteria from various sources in different locations from Sudan were tested for their insecticidal activity. The toxicity of these isolates ranged from 6.6 to 70% to the neonates of cotton bollworm, Helicoverpa armigera at 10 ppm concentration. The most effective ones are Kb-29, St-6 and Wh-1 comparable with HD-1. Toxicity of isolates to larvae of the red flour beetle, Tribolium castaneum ranged from 20 to 100%. Isolates St-2 and St-23 gave 100% larval mortality within 15 days of exposure and were at par with Ab-8, Ab-12, Kb-26, Kb-30, Om-4, Po-2, Po-5, Po-7, Sa-8 and Wh-5 and were also comparable with E. coli clone expressing Cry3 toxin. The most effective five isolates viz., Kb-29, St-2, St-6, St-23 and Wh-1 belonged to B. thuringiensis. The St-6 isolate, which also showed high toxicity to T. castaneum larvae, had cry1 genes along with coleopteran active cry28 genes, but not cry3 genes. Of the 25 isolates characterized with 16s DNA sequencing, seven belonged to Paenibacillus spp., one Lysinibacillus sphaericus, one Bacillus pumilus, four Bacillus spp., and rest 12 belonged to B. thuringiensis. Biochemical characterization in each species showed variation. The present study shows potential of some isolates like Kb-29, St-2, St-6, St-23 and Wh-1 as promising bioinsecticides.

  15. A novel bio-engineering approach to generate an eminent surface-functionalized template for selective detection of female sex pheromone of Helicoverpa armigera

    PubMed Central

    Moitra, Parikshit; Bhagat, Deepa; Pratap, Rudra; Bhattacharya, Santanu

    2016-01-01

    Plant pests exert serious effects on food production due to which the global crop yields are reduced by ~20–40 percent per year. Hence to meet the world’s food needs, loses of food due to crop pests must be reduced. Herein the silicon dioxide based MEMS devices are covalently functionalized for robust and efficient optical sensing of the female sex pheromones of the pests like Helicoverpa armigera for the first time in literature. The functionalized devices are also capable of selectively measuring the concentration of this pheromone at femtogram level which is much below the concentration of pheromone at the time of pest infestation in an agricultural field. Experiments are also performed in a confined region in the presence of male and female pests and tomato plants which directly mimics the real environmental conditions. Again the reversible use and absolutely trouble free transportation of these pheromone nanosensors heightens their potentials for commercial use. Overall, a novel and unique approach for the selective and reversible sensing of female sex pheromones of certain hazardous pests is reported herein which may be efficiently and economically carried forward from the research laboratory to the agricultural field. PMID:27892521

  16. No evidence for change in oviposition behaviour of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) after widespread adoption of transgenic insecticidal cotton.

    PubMed

    Zalucki, M P; Cunningham, J P; Downes, S; Ward, P; Lange, C; Meissle, M; Schellhorn, N A; Zalucki, J M

    2012-08-01

    Cotton growing landscapes in Australia have been dominated by dual-toxin transgenic Bt varieties since 2004. The cotton crop has thus effectively become a sink for the main target pest, Helicoverpa armigera. Theory predicts that there should be strong selection on female moths to avoid laying on such plants. We assessed oviposition, collected from two cotton-growing regions, by female moths when given a choice of tobacco, cotton and cabbage. Earlier work in the 1980s and 1990s on populations from the same geographic locations indicated these hosts were on average ranked as high, mid and low preference plants, respectively, and that host rankings had a heritable component. In the present study, we found no change in the relative ranking of hosts by females, with most eggs being laid on tobacco, then cotton and least on cabbage. As in earlier work, some females laid most eggs on cotton and aspects of oviposition behaviour had a heritable component. Certainly, cotton is not avoided as a host, and the implications of these finding for managing resistance to Bt cotton are discussed.

  17. Trans-generational desensitization and within-generational resensitization of a sucrose-best neuron in the polyphagous herbivore Helicoverpa armigera (Lepidoptera: Noctuidae)

    PubMed Central

    Ma, Ying; Li, Jingjing; Tang, Qingbo; Zhang, Xuening; Zhao, Xincheng; Yan, Fengming; van Loon, Joop J. A.

    2016-01-01

    Dietary exposure of insects to a feeding deterrent substance for hours to days can induce habituation and concomitant desensitization of the response of peripheral gustatory neurons to such a substance. In the present study, larvae of the herbivore Helicoverpa armigera were fed on diets containing either a high, medium or low concentration of sucrose, a major feeding stimulant. The responsiveness of the sucrose-best neuron in the lateral sensilla styloconica on the galea was quantified. Results showed the response of the sucrose-best neuron exposed to high-sucrose diets decreased gradually over successive generations, resulting in complete desensitization in the 5th and subsequent generations. However, the sensitivity was completely restored in the ninth generation after neonate larvae were exposed to low-sucrose diet. These findings demonstrate phenotypic plasticity and exclude inadvertent artificial selection for low sensitivity to sucrose. No significant changes were found in the sensitivity of caterpillars which experienced low- or medium-sucrose diets over the same generations. Such desensitization versus re-sensitization did not generalise to the phagosimulant myo-inositol-sensitive neuron or the feeding deterrent-sensitive neuron. Our results demonstrate that under conditions of high sucrose availability trans-generational desensitization of a neuron sensitive to this feeding stimulant becomes more pronounced whereas re-sensitization occurs within one generation. PMID:27966640

  18. Silencing of ecdysone receptor, insect intestinal mucin and sericotropin genes by bacterially produced double-stranded RNA affects larval growth and development in Plutella xylostella and Helicoverpa armigera.

    PubMed

    Israni, B; Rajam, M V

    2017-04-01

    RNA interference mediated gene silencing, which is triggered by double-stranded RNA (dsRNA), has become a important tool for functional genomics studies in various systems, including insects. Bacterially produced dsRNA employs the use of a bacterial strain lacking in RNaseIII activity and harbouring a vector with dual T7 promoter sites, which allow the production of intact dsRNA molecules. Here, we report an assessment of the functional relevance of the ecdysone receptor, insect intestinal mucin and sericotropin genes through silencing by dsRNA in two lepidopteran insect pests, Helicoverpa armigera and Plutella xylostella, both of which cause serious crop losses. Oral feeding of dsRNA led to significant reduction in transcripts of the target insect genes, which caused significant larval mortality with various moulting anomalies and an overall developmental delay. We also found a significant decrease in reproductive potential in female moths, with a drop in egg laying and compromised egg hatching from treated larvae as compared to controls. dsRNA was stable in the insect gut and was efficiently processed into small interfering RNAs (siRNAs), thus accounting for the phenotypes observed in the present work. The study revealed the importance of these genes in core insect processes, which are essential for insect development and survival.

  19. Overexpression of a Weed (Solanum americanum) Proteinase Inhibitor in Transgenic Tobacco Results in Increased Glandular Trichome Density and Enhanced Resistance to Helicoverpa armigera and Spodoptera litura

    PubMed Central

    Luo, Ming; Wang, Zhaoyu; Li, Huapeng; Xia, Kuai-Fei; Cai, Yinpeng; Xu, Zeng-Fu

    2009-01-01

    In this study we produced transgenic tobacco plants by overexpressing a serine proteinase inhibitor gene, SaPIN2a, from the American black nightshade Solanum americanum under the control of the CaMV 35S promoter using Agrobacterium tumefaciens-mediated transformation. SaPIN2a was properly transcribed and translated as indicated by Northern blot and Western blot analyses. Functional integrity of SaPIN2a in transgenic plants was confirmed by proteinase inhibitory activity assay. Bioassays for insect resistance showed that SaPIN2a-overexpressing transgenic tobacco plants were more resistant to cotton bollworm (Helicoverpa armigera) and tobacco cutworm (Spodoptera litura) larvae, two devastating pests of important crop plants, than the control plants. Interestingly, overexpression of SaPIN2a in transgenic tobacco plants resulted in a significant increase in glandular trichome density and a promotion of trichome branching, which could also provide an additional resistance mechanism in transgenic plants against insect pests. Therefore, SaPIN2a could be used as an alternative proteinase inhibitor for the production of insect-resistant transgenic plants. PMID:19468345

  20. Quantitative Analysis of Fitness Costs Associated with the Development of Resistance to the Bt Toxin Cry1Ac in Helicoverpa armigera

    PubMed Central

    Cao, Guangchun; Feng, Hongqiang; Guo, Fang; Wu, Kongming; Li, Xianchun; Liang, Gemei; Desneux, Nicolas

    2014-01-01

    Transgenic Bacillus thuringiensis (Bt) crops play an increasing role in pest control, and resistance management is a major issue in large-scale cultivation of Bt crops. The fitness cost of resistance in targeted pests is considered to be one of the main factors delaying resistance when using the refuge strategy. By comparing 10 resistant Helicoverpa armigera (Hubner) strains, showing various resistance levels to Bt toxin (Cry1Ac), to a susceptible strain, we showed an increasing fitness cost corresponding with increasing levels of resistance. The relationship between overall fitness cost C and the resistance ratio Rr could be described by C = 24.47/(1 + exp([1.57 - Log10Rr]/0.2)). This model predicted that the maximum overall fitness cost would be ~24% (±5.22) in the strains with the highest resistance level. The overall fitness cost was closely linked to egg hatching rate, fecundity, emergence rate, larval survival rate, and developmental duration of adults. Among fitness components measured, fecundity was the most sensitive trait linked to the resistance selection. To integrate the results into simulation models would be valuable in evaluating how variation in fitness cost may influence the development of resistance in pest populations, thus helping to develop enhanced refuge strategies. PMID:25005122

  1. A novel bio-engineering approach to generate an eminent surface-functionalized template for selective detection of female sex pheromone of Helicoverpa armigera

    NASA Astrophysics Data System (ADS)

    Moitra, Parikshit; Bhagat, Deepa; Pratap, Rudra; Bhattacharya, Santanu

    2016-11-01

    Plant pests exert serious effects on food production due to which the global crop yields are reduced by ~20-40 percent per year. Hence to meet the world’s food needs, loses of food due to crop pests must be reduced. Herein the silicon dioxide based MEMS devices are covalently functionalized for robust and efficient optical sensing of the female sex pheromones of the pests like Helicoverpa armigera for the first time in literature. The functionalized devices are also capable of selectively measuring the concentration of this pheromone at femtogram level which is much below the concentration of pheromone at the time of pest infestation in an agricultural field. Experiments are also performed in a confined region in the presence of male and female pests and tomato plants which directly mimics the real environmental conditions. Again the reversible use and absolutely trouble free transportation of these pheromone nanosensors heightens their potentials for commercial use. Overall, a novel and unique approach for the selective and reversible sensing of female sex pheromones of certain hazardous pests is reported herein which may be efficiently and economically carried forward from the research laboratory to the agricultural field.

  2. A P-glycoprotein gene serves as a component of the protective mechanisms against 2-tridecanone and abamectin in Helicoverpa armigera.

    PubMed

    Xiang, Min; Zhang, Lei; Lu, Yao; Tang, Qiuling; Liang, Pei; Shi, Xueyan; Song, Dunlun; Gao, Xiwu

    2017-09-05

    P-glycoprotein (P-gp) exists in animals, fungi and bacteria and likely evolved as a defense mechanism against harmful substances. Here a cDNA (4054bp) encoding a putative P-glycoprotein gene from Helicoverpa armigera was cloned and named HaPgp1. This putative HaPgp1 sequence encoded a protein of 1253 amino acids with a molecular mass of approximately 137kDa. qPCR analyses demonstrated that the expression of HaPgp1 was significantly higher in 4th instar larvae when compared to other developmental stages. HaPgp1 transcripts were more abundant in the head and fat bodies than in other tissues. Compared with the control, the expression of HaPgp1 reach a peak at 12h after the treatment by 2-tridecanone in all tissues. However, the expression of HaPgp1 increased from 12h to 48h after treatment with abamectin in all tissues. Immunohistochemistry analyses also verified that 2-tridecanone and abamectin can induce the increase of HaPgp1 expression. RNAi of HaPgp1 significantly raised the mortality rate of larvae treated by 2-tridecanone and abamectin, as compared to control larvae fed with GFP dsRNA. These results illustrate the possible involvement of HaPgp1 as a component of the protective mechanisms to plant secondary chemicals such as 2-tridecanone and to certain classes of insecticides, like abamectin. Copyright © 2017. Published by Elsevier B.V.

  3. Higher accumulation of proteinase inhibitors in flowers than leaves and fruits as a possible basis for differential feeding preference of Helicoverpa armigera on tomato (Lycopersicon esculentum Mill, Cv. Dhanashree).

    PubMed

    Damle, Mrunal S; Giri, Ashok P; Sainani, Mohini N; Gupta, Vidya S

    2005-11-01

    Tomato (Lycopersicon esculentum, Mill; cultivar- Dhanashree) proteinase inhibitors (PIs) were tested for their trypsin inhibitory (TI) and Helicoverpa armigera gut proteinases inhibitory (HGPI) activity in different organs of the tomato plants. Analysis of TI and HGPI distribution in various parts of the plant showed that flowers accumulated about 300 and 1000 times higher levels of TI while 700 and 400 times higher levels of HGPI as compared to those in leaves and fruits, respectively. Field observation that H. armigera larvae infest leaves and fruits but not the flowers could be at least partially attributed to the protective role-played by the higher levels of PIs in the flower tissue. Tomato PIs inhibited about 50-80% HGP activity of H. armigera larvae feeding on various host plants including tomato, of larvae exposed to non-host plant PIs and of various larval instars. Tomato PIs were found to be highly stable to insect proteinases wherein incubation of inhibitor with HGP even for 3h at optimum conditions did not affect inhibitory activity. Bioassay using H. armigera larvae fed on artificial diet containing tomato PIs revealed adverse effect on larval growth, pupae development, adult formation and fecundity.

  4. Effectiveness of two insect growth regulators against Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) and Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) and their impact on population densities of arthropod predators in cotton in Pakistan.

    PubMed

    Gogi, Muhammad D; Sarfraz, Rana M; Dosdall, Lloyd M; Arif, Muhammad J; Keddie, Andrew B; Ashfaq, Muhammad

    2006-10-01

    Field efficacies of two insect growth regulators (IGRs) at two recommended application rates, buprofezin at 370 and 555 g AI ha(-1) and lufenuron at 37 and 49 g AI ha(-1), were determined against the sweet potato whitefly, Bemisia tabaci (Gennadius), and the cotton bollworm, Helicoverpa armigera (Hübner), in experimental plots of cotton at the Directorate of Cotton Research, Faisalabad, Pakistan. Adverse effects of the IGRs on populations of associated arthropod predators, namely geocorids, chrysopids, coccinellids, formicids and arachnids, were also assessed. Both IGRs significantly reduced populations of B. tabaci at each application rate 24, 48 and 72 h after treatment, and higher doses were more effective than lower doses. Buprofezin was not effective against H. armigera at any tested dose for any time of treatment in any spray. Lufenuron applied at 37 and 49 g AI ha(-1) effectively suppressed H. armigera populations, resulting in significant reductions in crop damage. At lower doses, both IGRs appeared safe to predator populations, which did not differ significantly in IGR-treated versus untreated control plots. Population densities of formicids and coccinellids were significantly lower at high concentrations of both IGRs in treatment plots, possibly as a result of reduced prey availability. The potential role of buprofezin and lufenuron for control of B. tabaci and H. armigera in a spray programme and the likelihood of direct toxic effects of IGRs on predatory fauna of cotton are discussed.

  5. Effects of larval host plants on over-wintering preparedness and survival of the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

    PubMed

    Liu, Zhudong; Gong, Peiyu; Wu, Kujun; Wei, Wei; Sun, Jianghua; Li, Dianmo

    2007-10-01

    Laboratory colonies of cotton bollworm larvae, Helicoverpa armigera, kept at 20 degrees C under a photoperiod of L:D=10:14 were fed on five host plants (cotton, corn, kidney bean, tobacco and tomato) and an artificial diet (control) to determine the effects of larval host quality on survival and pupal over-wintering preparedness. A separate experiment showed that diapausing pupae weighed more and contained greater nutrient stores than did non-diapausing pupae. Diapausing pupae reared on different host plants showed significant differences in terms of over-wintering reserve storage, and degree of cold-hardiness (extent of low-molecular-weight substances and SCPs), and survivorship. The more nutrients the host plant had, the more the pupae weighed and the higher the levels of total lipids and glycogen. Body water content was also significantly affected by larval food quality. The mean pupal super-cooling capacities varied significantly from -16.7 to -18.9 degrees C according to host plants the larvae feed on, and these significantly related to water content, pupal weight, lipid and glycogen content, and the levels of glycerol. Levels of trehalose, glycerol, and inositol, which were mainly low-molecular-weight substances, showed no significant differences among different host plants, except for trehalose. Pupal mortality varied from 39.7% on corn to 3.3% on the artificial diet, which was significantly related to pupal weight, total lipid content, trehalose levels, and super-cooling points. These results suggest that larval food quality can affect survival and influence the over-wintering preparedness of the cotton bollworm.

  6. The Use of F2 Screening for Detection of Resistance to Emamectin Benzoate, Chlorantraniliprole, and Indoxacarb in Australian Populations of Helicoverpa armigera (Lepidoptera: Noctuidae).

    PubMed

    Bird, L J; Drynan, L J; Walker, P W

    2017-04-01

    The ability to effectively detect changes in susceptibility to insecticides is an integral component of resistance management strategies and is highly dependent upon precision of methods deployed. Between 2013 and 2016, F2 screens were performed for detection of resistance alleles in Helicoverpa armigera (Hübner) to emamectin benzoate, chlorantraniliprole, and indoxacarb in major cropping regions of eastern Australia. Resistance to emamectin benzoate was not detected. There were low but detectable levels of survival at discriminating concentrations of chlorantraniliprole and indoxacarb. Alleles conferring an advantage to chlorantraniliprole were present at a frequency of 0.0027 (95% CI 0.0012-0.0064; n = 1,817). Alleles conferring an advantage to indoxacarb were present at a frequency of 0.027 (95% CI 0.020-0.035; n = 1,863). Complementation tests for allelism in six of seven positive indoxacarb tests indicated that resistance was due to alleles present at the same locus. The majority (88%) of lines that tested positive for indoxacarb resistance deviated from a model of recessive inheritance. Pheromone-caught male moths contributed significantly greater numbers of F2 lines compared with moths derived from field-collected eggs or larvae. There was no difference in the detectability of indoxacarb resistance in F2 lines from pheromone-caught moths compared with moths derived from immature stages collected from the field and reared to adult under laboratory conditions. Therefore, we recommend the use of pheromone traps for sourcing insects for F2 screening as a more cost- and time-efficient alternative to traditional methods of sampling. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  7. The movement and distribution of Helicoverpa armigera (Hübner) larvae on pea plants is affected by egg placement and flowering.

    PubMed

    Perkins, L E; Cribb, B W; Hanan, J; Zalucki, M P

    2010-10-01

    The distribution and movement of 1st instar Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) larvae on whole garden pea (Pisum sativum L.) plants were determined in glasshouse trials. This economically-important herbivore attacks a wide variety of agricultural, horticultural and indigenous plants. To investigate the mechanisms underlying larval intra-plant movement, we used early-flowering and wild-type plant genotypes and placed eggs at different vertical heights within the plants, one egg per plant. Leaf water and nitrogen content and cuticle hardness were measured at the different plant heights. Of 92 individual larvae, 41% did not move from the node of eclosion, 49% moved upwards and 10% moved downwards with the distance moved being between zero and ten plant nodes. Larvae from eggs placed on the lower third of the plant left the natal leaf more often and moved further than larvae from eggs placed in the middle or upper thirds. The low nutritive value of leaves was the most likely explanation for more movement away from lower plant regions. Although larvae on flowering plants did not move further up or down than larvae on non-flowering plants, they more often departed the leaflet (within a leaf) where they eclosed. The final distribution of larvae was affected by plant genotype, with larvae on flowering plants found less often on leaflets and more often on stipules, tendrils and reproductive structures. Understanding intra-plant movement by herbivorous insects under natural conditions is important because such movement determines the value of economic loss to host crops. Knowing the behaviour underlying the spatial distribution of herbivores on plants will assist us to interpret field data and should lead to better informed pest management decisions.

  8. A Comparison of the Life-History Traits between Diapause and Direct Development iNdividuals in the Cotton Bollworm, Helicoverpa armigera

    PubMed Central

    Chen, Chao; Xia, Qin-Wen; Xiao, Hai-Jun; Xiao, Liang; Xue, Fang-Sen

    2014-01-01

    In order to understand the differences of life-history traits between diapause and direct development individuals in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), the development time, body size, growth rate, and adult longevity were investigated between the two populations, which were induced under 12:12 L:D and 16:8 L:D photoperiods, respectively, at 20, 22, and 25°C. The results indicated that the larval development time, pupal weight, adult weight, and growth rate were significantly different between diapause and direct developing individuals. The diapause developing individuals had a significantly higher pupal and adult weight and a longer larval time compared with direct developing individuals. However, the growth rate in diapause developing individuals was lower than that in the direct developing individuals. Analysis by GLM showed that larval time, pupal and adult weight, and growth rate were significantly influenced by both temperature and developmental pathway. The pupal and adult weights were greater in males than females in both developmental pathways, exhibiting sexual size dimorphism. The dimorphism in adult weight was more pronounced than in pupal weight because female pupae lost more weight at metamorphosis compared to male pupae. Protogyny was observed in both developmental pathways. However, the protogyny phenomenon was more pronounced at lower temperatures in direct developing individuals, whereas it was more pronounced in diapause developing individuals when they experienced higher temperatures in their larval stage and partial pupal period. The adult longevity of diapause developing individuals was significantly longer than that of direct developing individuals. The results reveal that the lifehistory strategy was different between diapause and direct developing individuals. PMID:25373166

  9. Global Metabolomic Analyses of the Hemolymph and Brain during the Initiation, Maintenance, and Termination of Pupal Diapause in the Cotton Bollworm, Helicoverpa armigera

    PubMed Central

    Lu, Yu-Xuan; Zhang, Qi; Xu, Wei-Hua

    2014-01-01

    A strategy known as diapause (developmental arrest) has evolved in insects to increase their survival rate under harsh environmental conditions. Diapause causes a dramatic reduction in the metabolic rate and drastically extends lifespan. However, little is known about the mechanisms underlying the metabolic changes involved. Using gas chromatography-mass spectrometry, we compared the changes in the metabolite levels in the brain and hemolymph of nondiapause- and diapause-destined cotton bollworm, Helicoverpa armigera, during the initiation, maintenance, and termination of pupal diapause. A total of 55 metabolites in the hemolymph and 52 metabolites in the brain were detected. Of these metabolites, 21 and 12 metabolite levels were altered in the diapause pupal hemolymph and brain, respectively. During diapause initiation and maintenance, the number of metabolites with increased levels in the hemolymph of the diapausing pupae is far greater than the number in the nondiapause pupae. These increased metabolites function as an energy source, metabolic intermediates, and cryoprotectants. The number of metabolites with decreased levels in the brain of diapausing pupae is far greater than the number in the nondiapause pupae. Low metabolite levels are likely to directly or indirectly repress the brain metabolic activity. During diapause termination, most of the metabolite levels in the hemolymph of the diapausing pupae rapidly decrease because they function as energy and metabolic sources that promote pupa-adult development. In conclusion, the metabolites with altered levels in the hemolymph and brain serve as energy and metabolic resources and help to maintain a low brain metabolic activity during diapause. PMID:24926789

  10. Impact of the Stem Extract of Thevetia neriifolia on the Feeding Potential and Histological Architecture of the Midgut Epithelial Tissue of Early Fourth Instars of Helicoverpa armigera Hübner.

    PubMed

    Mishra, Monika; Gupta, Kamal Kumar; Kumar, Sarita

    2015-01-01

    Helicoverpa armigera Hübner is one of the most important agricultural crop pests in the world causing heavy crop yield losses. The continued and indiscriminate use of synthetic insecticides in agriculture for their control has received wide public apprehension because of multifarious problems, including insecticide resistance, resurgence of pest species, environmental pollution, and toxic hazards to humans and nontarget organisms. These problems have necessitated the need to explore and develop alternative strategies using eco-friendly and biodegradable plant products. In view of this, the efficacy of Thevetia neriifolia methanol stem extract was evaluated against the early fourth instars of H. armigera as an antifeedant and stomach poison agent. Feeding of larvae with the diet containing 0.005%-5.0% extract resulted in 2.06%-37.35% antifeedant index; the diet with 5.0% extract caused 54.3% reduced consumption. The negative impact of extract on larval feeding resulted in 37.5%-77.7% starvation, causing adverse effects on the larval weight. Choice between control and experimental diet resulted in feeding preference of larvae for the control diet, leading to 7.3%-42.9% reduced consumption of extract-containing diet. The only exception was the diet with 0.005% extract, which could not cause any deterrence. The midgut histological architecture of H. armigera larvae fed with 0.005%-0.05% extract-containing diet with negligible antifeedant potential showed significant damage, shrinkage, and distortion and vacuolization of gut tissues and peritrophic membrane, causing the disintegration of epithelial, goblet, and regenerative cells; the damage increased with the increase in concentration. These changes in the gut caused negative impact on the digestion and absorption of food and thus nutritional deficiency in the larvae, which could probably affect their growth and development. This study reveal the appreciable stomach poison potential of T. neriifolia stem methanol

  11. Impact of the Stem Extract of Thevetia neriifolia on the Feeding Potential and Histological Architecture of the Midgut Epithelial Tissue of Early Fourth Instars of Helicoverpa armigera Hübner

    PubMed Central

    Mishra, Monika; Gupta, Kamal Kumar; Kumar, Sarita

    2015-01-01

    Helicoverpa armigera Hübner is one of the most important agricultural crop pests in the world causing heavy crop yield losses. The continued and indiscriminate use of synthetic insecticides in agriculture for their control has received wide public apprehension because of multifarious problems, including insecticide resistance, resurgence of pest species, environmental pollution, and toxic hazards to humans and nontarget organisms. These problems have necessitated the need to explore and develop alternative strategies using eco-friendly and biodegradable plant products. In view of this, the efficacy of Thevetia neriifolia methanol stem extract was evaluated against the early fourth instars of H. armigera as an antifeedant and stomach poison agent. Feeding of larvae with the diet containing 0.005%–5.0% extract resulted in 2.06%–37.35% antifeedant index; the diet with 5.0% extract caused 54.3% reduced consumption. The negative impact of extract on larval feeding resulted in 37.5%–77.7% starvation, causing adverse effects on the larval weight. Choice between control and experimental diet resulted in feeding preference of larvae for the control diet, leading to 7.3%–42.9% reduced consumption of extract-containing diet. The only exception was the diet with 0.005% extract, which could not cause any deterrence. The midgut histological architecture of H. armigera larvae fed with 0.005%–0.05% extract-containing diet with negligible antifeedant potential showed significant damage, shrinkage, and distortion and vacuolization of gut tissues and peritrophic membrane, causing the disintegration of epithelial, goblet, and regenerative cells; the damage increased with the increase in concentration. These changes in the gut caused negative impact on the digestion and absorption of food and thus nutritional deficiency in the larvae, which could probably affect their growth and development. This study reveal the appreciable stomach poison potential of T. neriifolia stem

  12. Diversity of aminopeptidases, derived from four lepidopteran gene duplications, and polycalins expressed in the midgut of Helicoverpa armigera: Identification of proteins binding the δ-endotoxin, Cry1Ac of Bacillus thuringiensis

    PubMed Central

    Angelucci, Constanza; Barrett-Wilt, Gregory A.; Hunt, Donald F.; Akhurst, Raymond J.; East, Peter D.; Gordon, Karl H.J.; Campbell, Peter M.

    2010-01-01

    Helicoverpa armigera midgut proteins that bind the Bacillus thuringiensis (Bt) δ-endotoxin Cry1Ac were purified by affinity chromatography. SDS-PAGE showed that several proteins were eluted with N-acetylgalactosamine and no further proteins were detected after elution with urea. Tandem mass spectral data for tryptic peptides initially indicated that the proteins resembled aminopeptidases (APNs) from other lepidopterans and cDNA sequences for seven APNs were isolated from H. armigera through a combination of cloning with primers derived from predicted peptide sequences and established EST libraries. Phylogenetic analysis showed lepidopteran APN genes in nine clades of which five were part of a lepidopteran-specific radiation. The Cry1Ac-binding proteins were then identified with four of the seven HaAPN genes. Three of those four APNs are likely orthologs of APNs characterised as Cry1Ac-binding proteins in other lepidopterans. The fourth Cry1Ac-binding APN has orthologs not previously identified as Cry1Ac-binding partners. The HaAPN genes were expressed predominantly in the midgut through larval development. Each showed consistent expression along the length of the midgut but five of the genes were expressed at levels about two orders of magnitude greater than the remaining two. The remaining mass spectral data identified sequences encoding polycalin proteins with multiple lipocalin-like domains. A polycalin has only been previously reported in another lepidopteran, Bombyx mori, but polycalins in both species are now linked with binding of Bt Cry toxins. This is the first report of hybrid, lipocalin-like domains in shorter polycalin sequences that are not present in the longest sequence. We propose that these hybrid domains are generated by alternative splicing of the mRNA. PMID:18549954

  13. Expression of a Chimeric Gene Encoding Insecticidal Crystal Protein Cry1Aabc of Bacillus thuringiensis in Chickpea (Cicer arietinum L.) Confers Resistance to Gram Pod Borer (Helicoverpa armigera Hubner.).

    PubMed

    Das, Alok; Datta, Subhojit; Thakur, Shallu; Shukla, Alok; Ansari, Jamal; Sujayanand, G K; Chaturvedi, Sushil K; Kumar, P A; Singh, N P

    2017-01-01

    Domain swapping and generation of chimeric insecticidal crystal protein is an emerging area of insect pest management. The lepidopteran insect pest, gram pod borer (Helicoverpa armigera H.) wreaks havoc to chickpea crop affecting production. Lepidopteran insects were reported to be controlled by Bt (cryI) genes. We designed a plant codon optimized chimeric Bt gene (cry1Aabc) using three domains from three different cry1A genes (domains I, II, and III from cry1Aa, cry1Ab, and cry1Ac, respectively) and expressed it under the control of a constitutive promoter in chickpea (cv. DCP92-3) to assess its effect on gram pod borer. A total of six transgenic chickpea shoots were established by grafting into mature fertile plants. The in vitro regenerated (organogenetic) shoots were selected based on antibiotic kanamycin monosulfate (100 mg/L) with transformation efficiency of 0.076%. Three transgenic events were extensively studied based on gene expression pattern and insect mortality across generations. Protein expression in pod walls, immature seeds and leaves (pre- and post-flowering) were estimated and expression in pre-flowering stage was found higher than that of post-flowering. Analysis for the stable integration, expression and insect mortality (detached leaf and whole plant bioassay) led to identification of efficacious transgenic chickpea lines. The chimeric cry1Aabc expressed in chickpea is effective against gram pod borer and generated events can be utilized in transgenic breeding program.

  14. Prevalence of cry2-type genes in Bacillus thuringiensis isolates recovered from diverse habitats in India and isolation of a novel cry2Af2 gene toxic to Helicoverpa armigera (cotton boll worm).

    PubMed

    Katara, Jawahar Lal; Kaur, Sarvjeet; Kumari, Gouthami Krishna; Singh, Nagendra Kumar

    2016-12-01

    Insecticidal cry and vip genes from Bacillus thuringiensis (Bt) have been used for control of lepidopteran insects in transgenic crops. However, novel genes are required for gene pyramiding to delay evolution of resistance to the currently deployed genes. Two PCR-based techniques were employed for screening of cry2-type genes in 129 Bt isolates from diverse habitats in India and 27 known Bt strains. cry2Ab-type genes were more prevalent than cry2Aa- and cry2Ac-type genes. Correlation between source of isolates and abundance of cry2-type genes was not observed. Full-length cry2A-type genes were amplified by PCR from 9 Bt isolates and 4 Bt strains. The genes from Bt isolates SK-758 from Sorghum grain dust and SK-793 from Chilli seeds warehouse, Andhra Pradesh, were cloned and sequenced. The gene from SK-758 (NCBI GenBank accession No. GQ866915) was novel, while that from SK-793 (NCBI GenBank accession No. GQ866914) was identical to the cry2Ab1 gene. The Bacillus thuringiensis Nomenclature Committee ( http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/toxins2.html ) named these genes cry2Af2 and cry2Ab16, respectively. The cry2Af2 gene was expressed in Escherichia coli and found to be toxic towards Helicoverpa armigera. The cry2Af2 gene will be useful for pyramiding in transgenic crops.

  15. Expression of a Chimeric Gene Encoding Insecticidal Crystal Protein Cry1Aabc of Bacillus thuringiensis in Chickpea (Cicer arietinum L.) Confers Resistance to Gram Pod Borer (Helicoverpa armigera Hubner.)

    PubMed Central

    Das, Alok; Datta, Subhojit; Thakur, Shallu; Shukla, Alok; Ansari, Jamal; Sujayanand, G. K.; Chaturvedi, Sushil K.; Kumar, P. A.; Singh, N. P.

    2017-01-01

    Domain swapping and generation of chimeric insecticidal crystal protein is an emerging area of insect pest management. The lepidopteran insect pest, gram pod borer (Helicoverpa armigera H.) wreaks havoc to chickpea crop affecting production. Lepidopteran insects were reported to be controlled by Bt (cryI) genes. We designed a plant codon optimized chimeric Bt gene (cry1Aabc) using three domains from three different cry1A genes (domains I, II, and III from cry1Aa, cry1Ab, and cry1Ac, respectively) and expressed it under the control of a constitutive promoter in chickpea (cv. DCP92-3) to assess its effect on gram pod borer. A total of six transgenic chickpea shoots were established by grafting into mature fertile plants. The in vitro regenerated (organogenetic) shoots were selected based on antibiotic kanamycin monosulfate (100 mg/L) with transformation efficiency of 0.076%. Three transgenic events were extensively studied based on gene expression pattern and insect mortality across generations. Protein expression in pod walls, immature seeds and leaves (pre- and post-flowering) were estimated and expression in pre-flowering stage was found higher than that of post-flowering. Analysis for the stable integration, expression and insect mortality (detached leaf and whole plant bioassay) led to identification of efficacious transgenic chickpea lines. The chimeric cry1Aabc expressed in chickpea is effective against gram pod borer and generated events can be utilized in transgenic breeding program. PMID:28871265

  16. Characterization of the resistance to Vip3Aa in Helicoverpa armigera from Australia and the role of midgut processing and receptor binding

    PubMed Central

    Chakroun, Maissa; Banyuls, Núria; Walsh, Tom; Downes, Sharon; James, Bill; Ferré, Juan

    2016-01-01

    Crops expressing genes from Bacillus thuringiensis (Bt crops) are among the most successful technologies developed for the control of pests but the evolution of resistance to them remains a challenge. Insect resistant cotton and maize expressing the Bt Vip3Aa protein were recently commercialized, though not yet in Australia. We found that, although relatively high, the frequency of alleles for resistance to Vip3Aa in field populations of H. armigera in Australia did not increase over the past four seasons until 2014/15. Three new isofemale lines were determined to be allelic with previously isolated lines, suggesting that they belong to one common gene and this mechanism is relatively frequent. Vip3Aa-resistance does not confer cross-resistance to Cry1Ac or Cry2Ab. Vip3Aa was labeled with 125I and used to show specific binding to H. armigera brush-border membrane vesicles (BBMV). Binding was of high affinity (Kd = 25 and 19 nM for susceptible and resistant insects, respectively) and the concentration of binding sites was high (Rt = 140 pmol/mg for both). Despite the narrow-spectrum resistance, binding of 125I-labeled Vip3Aa to BBMV of resistant and susceptible insects was not significantly different. Proteolytic conversion of Vip3Aa protoxin into the activated toxin rendered the same products, though it was significantly slower in resistant insects. PMID:27095284

  17. Characterization of the resistance to Vip3Aa in Helicoverpa armigera from Australia and the role of midgut processing and receptor binding.

    PubMed

    Chakroun, Maissa; Banyuls, Núria; Walsh, Tom; Downes, Sharon; James, Bill; Ferré, Juan

    2016-04-20

    Crops expressing genes from Bacillus thuringiensis (Bt crops) are among the most successful technologies developed for the control of pests but the evolution of resistance to them remains a challenge. Insect resistant cotton and maize expressing the Bt Vip3Aa protein were recently commercialized, though not yet in Australia. We found that, although relatively high, the frequency of alleles for resistance to Vip3Aa in field populations of H. armigera in Australia did not increase over the past four seasons until 2014/15. Three new isofemale lines were determined to be allelic with previously isolated lines, suggesting that they belong to one common gene and this mechanism is relatively frequent. Vip3Aa-resistance does not confer cross-resistance to Cry1Ac or Cry2Ab. Vip3Aa was labeled with (125)I and used to show specific binding to H. armigera brush-border membrane vesicles (BBMV). Binding was of high affinity (Kd = 25 and 19 nM for susceptible and resistant insects, respectively) and the concentration of binding sites was high (Rt = 140 pmol/mg for both). Despite the narrow-spectrum resistance, binding of (125)I-labeled Vip3Aa to BBMV of resistant and susceptible insects was not significantly different. Proteolytic conversion of Vip3Aa protoxin into the activated toxin rendered the same products, though it was significantly slower in resistant insects.

  18. Coexpression of the silent cry2Ab27 together with cry1 genes in Bacillus thuringiensis subsp. aizawai SP41 leads to formation of amorphous crystal toxin and enhanced toxicity against Helicoverpa armigera.

    PubMed

    Somwatcharajit, Rasapirose; Tiantad, Itsares; Panbangred, Watanalai

    2014-02-01

    The unexpressed cry2Ab27 gene of Bacillus thuringiensis subsp. aizawai SP41 (SP41) consists of a single open reading frame (ORF) of 1902bp encoding for 634 amino acid residues. The cry2Ab27 gene appears to be silent due to the lack of promoter and terminator sequences. In this study we fused the cry2Ab27 ORF with the cry1Ab promoter (500bp) and the terminator (300bp) in vector pHT304-18Z in order to drive the expression of cry2Ab27 in both SP41 and an acrystaliferous, B. thuringiensis subsp. thuringiensis 407 (407). A protein with a molecular mass of 65kDa, consistent with the Cry2Ab protein, was detected in both transformants using SDS-PAGE and Western blot analysis. Bipyramidal crystals were observed in SP41 and its transformant containing the pHT304-18Z vector (SPHT) in contrast, cells expressing cry2Ab27 (SPC2) exhibited crystal proteins with irregular shapes. No inclusion protein was detected in the 407 transformant expressing the cry2Ab27 gene. Cry2Ab27 was found in the purified crystal toxin from strain SPC2. The solubilized crystal toxin proteins from SPC2 were 6.9-fold more toxic toward the larvae of Helicoverpa armigera compared to toxin proteins from SPHT. However SPC2 crystal toxin displayed only slightly higher toxicity against the larvae of Spodoptera litura and S. exigua compared to SPHT produced toxin. Our data support the use of Cry2Ab in combination with the Cry1 toxin for enhanced control of heliothine insect pests.

  19. Proteomic Analysis of Mamestra Brassicae Nucleopolyhedrovirus Progeny Virions from Two Different Hosts

    PubMed Central

    Hou, Dianhai; Chen, Xi

    2016-01-01

    Mamestra brassicae nucleopolyhedrovirus (MabrNPV) has a wide host range replication in more than one insect species. In this study, a sequenced MabrNPV strain, MabrNPV-CTa, was used to perform proteomic analysis of both BVs and ODVs derived from two infected hosts: Helicoverpa armigera and Spodoptera exigua. A total of 82 and 39 viral proteins were identified in ODVs and BVs, respectively. And totally, 23 and 76 host proteins were identified as virion-associated with ODVs and BVs, respectively. The host proteins incorporated into the virus particles were mainly involved in cytoskeleton, signaling, vesicle trafficking, chaperone and metabolic systems. Some host proteins, such as actin, cyclophilin A and heat shock protein 70 would be important for viral replication. Several host proteins involved in immune response were also identified in BV, and a C-type lectin protein was firstly found to be associated with BV and its family members have been demonstrated to be involved in entry process of other viruses. This study facilitated the annotation of baculovirus genome, and would help us to understand baculovirus virion structure. Furthermore, the identification of host proteins associated with virions produced in vivo would facilitate investigations on the involvement of intriguing host proteins in virus replication. PMID:27058368

  20. Both Lymantria dispar Nucleopolyhedrovirus Enhancin Genes Contribute to Viral Potency

    PubMed Central

    Popham, Holly J. R.; Bischoff, David S.; Slavicek, James M.

    2001-01-01

    Enhancins are a group of proteins first identified in granuloviruses (GV) that have the ability to enhance nuclear polyhedrosis virus potency. We had previously identified an enhancin gene (E1) in the Lymantria dispar multinucleocapsid nucleopolyhedrovirus (LdMNPV) (D. S. Bischoff and J. M. Slavicek, J. Virol. 71:8133–8140, 1997). Inactivation of the E1 gene product within the viral genome lowered viral potency by an average of 2.9-fold. A second enhancin gene (E2) was identified when the entire genome of LdMNPV was sequenced (Kuzio et al., Virology 253:17–34, 1999). The E2 protein exhibits approximately 30% amino acid identity to the LdMNPV E1 protein as well as the enhancins from Trichoplusia ni GV, Pseudaletia unipuncta GV, Helicoverpa armigera GV, and Xestia c-nigrum GV. Northern analysis of viral RNA indicated that the E2 gene transcripts are expressed at late times postinfection from a consensus baculovirus late promoter. The effect of the enhancin proteins on viral potency was investigated through bioassay using two recombinant viruses, one with a deletion in the E2 gene (E2del) and a second with deletion mutations in both enhancin genes (E1delE2del). The enhancin gene viral constructs were verified by Southern analysis and shown not to produce enhancin gene transcripts by Northern analysis. The E2del virus exhibited an average decrease in viral potency of 1.8-fold compared to wild-type virus. In the same bioassays, the recombinant virus E1cat, which does not produce an E1 gene transcript, exhibited an average decrease in viral potency of 2.3-fold compared to control virus. The E1delE2del virus exhibited an average decrease in viral potency of 12-fold compared to wild-type virus. Collectively, these results suggest that both LdMNPV enhancin genes contribute to viral potency, that each enhancin protein can partially compensate for the lack of the other protein, and that both enhancin genes are necessary for wild-type viral potency. PMID:11507209

  1. Complete genomic sequences and comparative analysis of Mamestra brassicae nucleopolyhedrovirus isolated in Korea.

    PubMed

    Choi, Jae Bang; Heo, Won Il; Shin, Tae Young; Bae, Sung Min; Kim, Woo Jin; Kim, Ju Il; Kwon, Min; Choi, Jae Young; Je, Yeon Ho; Jin, Byung Rae; Woo, Soo Dong

    2013-08-01

    Mamestra brassicae nucleopolyhedrovirus-K1 (MabrNPV-K1) was isolated from naturally infected M. brassicae (Lepidoptera: Noctuidae) larvae in Korea. The full genome sequences of MabrNPV-K1 were determined, analysed and compared to those of other baculoviruses. The MabrNPV-K1 genome consisted of 152,710 bp and had an overall G + C content of 39.9%. Computer-assisted analysis predicted 158 open reading frames (ORFs) of 150 nucleotides or greater that showed minimal overlap. Two inhibitor of apoptosis (iap) and six baculovirus repeated ORFs were interspersed in the MabrNPV-K1 genome. The unique MabrNPV-K1 ORF133 was identified in the MabrNPV-K1 genome that was not previously reported in baculoviruses. The gene content and arrangement in MabrNPV-K1 had the highest similarity with those of Helicoverpa armigera MNPV (HearMNPV) and Mamestra configurata NPV-B (MacoNPV-B), and their shared homologous genes were 99% collinear. The MabrNPV-K1 genome contained four homologous repeat regions (hr1, hr2, hr3 and hr4) that accounted for 3.3% of the genome. The genomic positions of the four MabrNPV-K1 hr regions were conserved among those of HearMNPV and MacoNPV-B. The gene parity plot, percent identity of the gene homologues and a phylogenetic analysis suggested that these three viruses are closely related not only to each other but also to the same virus strains rather than different virus species.

  2. Efficacy of four traps in capturing male Helicoverpa moths in north Florida

    USDA-ARS?s Scientific Manuscript database

    Species in the genus Helicoverpa include agricultural pests that attack a variety of crops worldwide. In North America, the most important species is H. zea but in the Eastern Hemisphere H. armigera is the most important species and is a threat to invade the U.S. USDA-APHIS-PPQ has an active trapp...

  3. Efficacies of four pheromone-baited traps in capturing male Helicoverpa (Lepidoptera: Noctuidae) moths in northern Florida

    USDA-ARS?s Scientific Manuscript database

    Helicoverpa armigera (Hübner) is a serious pest of grain, row, and vegetable crops throughout much of the world, although it is currently not established in the United States. USDA-APHIS and the Cooperative Agricultural Pest Survey program are charged with the responsibility to monitor for this ins...

  4. Two Year Field Study to Evaluate the Efficacy of Mamestra brassicae Nucleopolyhedrovirus Combined with Proteins Derived from Xestia c-nigrum Granulovirus

    PubMed Central

    Goto, Chie; Mukawa, Shigeyuki; Mitsunaga, Takayuki

    2015-01-01

    Japan has only three registered baculovirus biopesticides despite its long history of studies on insect viruses. High production cost is one of the main hindrances for practical use of baculoviruses. Enhancement of insecticidal effect is one possible way to overcome this problem, so there have been many attempts to develop additives for baculoviruses. We found that alkaline soluble proteins of capsules (GVPs) of Xestia c-nigrum granulovirus can increase infectivity of some viruses including Mamestra brassicae nucleopolyhedrovirus (MabrNPV), and previously reported that MabrNPV mixed with GVPs was highly infectious to three important noctuid pests of vegetables in the following order, Helicoverpa armigera, M. brassicae, and Autographa nigrisigna. In this study, small-plot experiments were performed to assess concentrations of MabrNPV and GVPs at three cabbage fields and a broccoli field for the control of M. brassicae. In the first experiment, addition of GVPs (10 µg/mL) to MabrNPV at 106 OBs/mL resulted in a significant increase in NPV infection (from 53% to 66%). In the second experiment, the enhancing effect of GVP on NPV infection was confirmed at 10-times lower concentrations of MabrNPV. In the third and fourth experiments, a 50% reduction in GVPs (from 10 µg/mL to 5 µg/mL) did not result in a lowering of infectivity of the formulations containing MabrNPV at 105 OBs/mL. These results indicate that GVPs are promising additives for virus insecticides. PMID:25760139

  5. Evolution, ecology and management of resistance in Helicoverpa spp. to Bt cotton in Australia.

    PubMed

    Downes, Sharon; Mahon, Rod

    2012-07-01

    Prior to the widespread adoption of two-gene Bt cotton (Bollgard II®) in Australia, the frequency of resistance alleles to one of the deployed proteins (Cry2Ab) was at least 0.001 in the pests targeted namely, Helicoverpa armigera and Helicoverpa punctigera. In the 7 years hence, there has been a statistically significant increase in the frequency of alleles conferring Cry2Ab resistance in field populations of H. punctigera. This paper reviews the history of deploying Bt cotton in Australia, the characteristics of the isolated Cry2Ab resistance that likely impact on resistance evolution, aspects of the efficacy of Bollgard IIχ, and the behavioural ecology of Helicoverpa spp. larvae as it pertains to resistance management. It also presents up-to-date frequencies of resistant alleles for H. punctigera and reviews the same information for H. armigera. This is followed by a discussion of current resistance management strategies. The consequences of the imminent release of a third generation product that utilizes the novel vegetative insecticidal protein Vip3A are then considered. The area planted to Bt-crops is anticipated to continue to rise worldwide and many biotechnical companies intend to add Vip3A to existing products; therefore the information reviewed herein for Australia is likely to be pertinent to other situations.

  6. Evaluation of the natural refuge function for Helicoverpa arnigera (Lepidoptera: Noctuidae) within Bacillus thuringiensis transgenic cotton growing areas in north China.

    PubMed

    Wu, Kongming; Guo, Yuyuan; Gao, Shansong

    2002-08-01

    The density of Helicoverpa armigera (Hübner) populations on Bacillus thuringiensis Berliner (Bt) transgenic cotton, corn, peanut, and soybean; differences in its development on Bt cotton and common (nontransgenic) cotton; and the potential for mating among populations from Bt cotton fields and other crop fields were investigated in the suburbs of Xinxiang City (Henan Province) and Langfang City (Hebei Province) in the southern and northern parts of north China, respectively. Although development of H. armigera on Bt cotton was much slower than on common cotton, there was a still high probability of mating between populations from Bt cotton and other sources due to the scattered emergence pattern of H. armigera adults, and overlap of the second and third generations. In a cotton and corn growing region, early and late planted corn provided suitable refugia for the third and fourth generations of H. armigera, but not for the second generation. In a cotton and soybean/ peanut mix system, noncotton crops provided a natural refugia from the second- to fourth-generation H. armigera, but function of the refuge would closely depend on the proportion of Bt cotton. Consequently, it may be necessary to compensate the original mixed cropping patterns in different areas for delaying resistance development of H. armigera to Bt cotton.

  7. Sensillar expression and responses of olfactory receptors reveal different peripheral coding in two Helicoverpa species using the same pheromone components

    PubMed Central

    Chang, Hetan; Guo, Mengbo; Wang, Bing; Liu, Yang; Dong, Shuanglin; Wang, Guirong

    2016-01-01

    Male moths efficiently recognize conspecific sex pheromones thanks to their highly accurate and specific olfactory system. The Heliothis/Helicoverpa species are regarded as good models for studying the perception of sex pheromones. In this study, we performed a series of experiments to investigate the peripheral mechanisms of pheromone coding in two-closely related species, Helicoverpa armigera and H. assulta. The morphology and distribution patterns of sensilla trichoidea are similar between the two species when observed at the scanning electron microscope, but their performances are different. In H. armigera, three functional types of sensilla trichoidea (A, B and C) were found to respond to different pheromone components, while in H. assulta only two types of such sensilla (A and C) could be detected. The response profiles of all types of sensilla trichoidea in the two species well matched the specificities of the pheromone receptors (PRs) expressed in the same sensilla, as measured in voltage-clamp experiments. The expressions of PRs in neighboring olfactory sensory neurons (OSNs) within the same trichoid sensillum were further confirmed by in situ hybridization. Our results show how the same pheromone components can code for different messages at the periphery of two Helicoverpa species. PMID:26744070

  8. [Evaluation of reference genes for quantitative real-time PCR normalization in cotton bollworm, Helicoverna armigera].

    PubMed

    Chandra, G Sharath; Asokan, R; Manamohan, M; Kumar, N K K; Sita, T

    2014-01-01

    Reverse-transcription quantitative real-time PCR (RT-qPCR), a sensitive technique is being extensively employed in quantification of gene expression. However this requires normalization with suitable reference gene (RG) which is crucial in minimizing inter sample variations. Information regarding suitable RG is scarce in general and more so in insects, including the cotton bollworm, Helicoverpa armigera, an economically important pest. In management of this pest RNA interference (RNAi), is perceived as a potential tool, which is achieved by double-stranded RNA (dsRNA) delivery. These studies demand accurate quantification of gene silencing. In this study we assessed the suitability of five RGs viz. β-actin (ACTB), 18S rRNA (18S), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-tubulin (TUB) and elongation fator-1-alfa (EF1-α) for gene expression studies in dsRNA treatment and across different developmental stages of H. armigera and ranked using geNorm, NormFinder and BestKeeper software programs. Data analysis revealed that best ranked RGs were varied in dsRNA treatment and in developmental stages. Under dsRNA treatment, 18S and GAPDH were more stable whereas, TUB and GAPDH were more stable across developmental stages. We also demonstrate that inappropriate selection of RG led to erroneous estimation of the target gene, chymotrypsin, expression. These results facilitate accurate quantification of gene expression in H. armigera.

  9. High Susceptibility to Cry1Ac and Low Resistance Allele Frequency Reduce the Risk of Resistance of Helicoverpa armigers to Bt Soybean in Brazil.

    PubMed

    Dourado, Patrick M; Bacalhau, Fabiana B; Amado, Douglas; Carvalho, Renato A; Martinelli, Samuel; Head, Graham P; Omoto, Celso

    2016-01-01

    The Old World bollworm, Helicoverpa armigera (Hübner), was recently introduced into Brazil, where it has caused extensive damage to cotton and soybean crops. MON 87701 × MON 89788 soybean, which expresses the Bt protein Cry1Ac, was recently deployed in Brazil, providing high levels of control against H. armigera. To assess the risk of resistance to the Cry1Ac protein expressed by MON 87701 × MON 89788 soybean in Brazil, we conducted studies to evaluate the baseline susceptibility of H. armigera to Cry1Ac, in planta efficacy including the assessment of the high-dose criterion, and the initial resistance allele frequency based on an F2 screen. The mean Cry1Ac lethal concentration (LC50) ranged from 0.11 to 1.82 μg·mL-1 of diet among all H. armigera field populations collected from crop seasons 2013/14 to 2014/15, which indicated about 16.5-fold variation. MON 87701 × MON 89788 soybean exhibited a high level of efficacy against H. armigera and most likely met the high dose criterion against this target species in leaf tissue dilution bioassays up to 50 times. A total of 212 F2 family lines of H. armigera were established from field collections sampled from seven locations across Brazil and were screened for the presence of MON 87701 × MON 89788 soybean resistance alleles. None of the 212 families survived on MON 87701 × MON 89788 soybean leaf tissue (estimated allele frequency = 0.0011). The responses of H. armigera to Cry1Ac protein, high susceptibility to MON 87701 × MON 89788 soybean, and low frequency of resistance alleles across the main soybean-producing regions support the assumptions of a high-dose/refuge strategy. However, maintenance of reasonable compliance with the refuge recommendation will be essential to delay the evolution of resistance in H. armigera to MON 87701 × MON 89788 soybean in Brazil.

  10. A host-plant specialist, Helicoverpa assulta, is more tolerant to capsaicin from Capsicum annuum than other noctuid species.

    PubMed

    Ahn, Seung-Joon; Badenes-Pérez, Francisco R; Heckel, David G

    2011-09-01

    Plant secondary compounds not only play an important role in plant defense, but have been a driving force for host adaptation by herbivores. Capsaicin (8-methyl-N-vanillyl-6-nonenamide), an alkaloid found in the fruit of Capsicum spp. (Solanaceae), is responsible for the pungency of hot pepper fruits and is unique to the genus. The oriental tobacco budworm, Helicoverpa assulta (Lepidoptera: Noctuidae), is a specialist herbivore feeding on solanaceous plants including Capsicum annuum, and is one of a very few insect herbivores worldwide capable of feeding on hot pepper fruits. To determine whether this is due in part to an increased physiological tolerance of capsaicin, we compared H. assulta with another specialist on Solanaceae, Heliothis subflexa, and four generalist species, Spodoptera frugiperda, Heliothis virescens, Helicoverpa armigera, and Helicoverpa zea, all belonging to the family Noctuidae. When larvae were fed capsaicin-spiked artificial diet for the entire larval period, larval mortality increased in H. subflexa and H. zea but decreased in H. assulta. Larval growth decreased on the capsaicin-spiked diet in four of the species, was unaffected in H. armigera and increased in H. assulta. Food consumption and utilization experiments showed that capsaicin decreased relative consumption rate (RCR), relative growth rate (RGR) and approximate digestibility (AD) in H. zea, and increased AD and the efficiency of conversion of ingested food (ECI) in H. armigera; whereas it did not significantly change any of these nutritional indices in H. assulta. The acute toxicity of capsaicin measured by injection into early fifth instar larvae was less in H. assulta than in H. armigera and H. zea. Injection of high concentrations produced abdominal paralysis and self-cannibalism. Injection of sub-lethal doses of capsaicin resulted in reduced pupal weights in H. armigera and H. zea, but not in H. assulta. The results indicate that H. assulta is more tolerant to capsaicin than

  11. Biology, Ecology, and Evolving Management of Helicoverpa zea (Lepidoptera: Noctuidae) in Sweet Corn in the United States.

    PubMed

    Olmstead, Daniel L; Nault, Brian A; Shelton, Anthony M

    2016-08-01

    The corn earworm, Helicoverpa zea (Boddie), is a polyphagous pest found throughout the United States, where it attacks many field and vegetable crops. Although H. zea has long been a traditional pest of sweet corn, its importance to this crop has increased dramatically over the past two decades. In this review, we summarize information critical for current and future management of H. zea in sweet corn production in the United States. First, we discuss the pest status of H. zea and its life history, including migration, infestation and larval development, diapause, overwintering, and abiotic factors that affect its biology. Next we describe monitoring methods, crop protection decision-making processes, chemical control options, and the use of genetic technologies for control of H. zea Alternative H. zea management options including biological control, cultural controls, host plant resistance, and pheromone disruption are also reviewed. The role of climate change and its effects on H. zea and its ecology are discussed, as well as the recent invasion of its relative, Helicoverpa armigera (Hübner), which is a major pest of corn in other parts of the world. To conclude, we suggest future research opportunities for H. zea and H. armigera management in sweet corn. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  12. Specific binding of Bacillus thuringiensis Cry2A insecticidal proteins to a common site in the midgut of Helicoverpa species.

    PubMed

    Hernández-Rodríguez, Carmen Sara; Van Vliet, Adri; Bautsoens, Nadine; Van Rie, Jeroen; Ferré, Juan

    2008-12-01

    For a long time, it has been assumed that the mode of action of Cry2A toxins was unique and different from that of other three-domain Cry toxins due to their apparent nonspecific and unsaturable binding to an unlimited number of receptors. However, based on the homology of the tertiary structure among three-domain Cry toxins, similar modes of action for all of them are expected. To confirm this hypothesis, binding assays were carried out with (125)I-labeled Cry2Ab. Saturation assays showed that Cry2Ab binds in a specific and saturable manner to brush border membrane vesicles (BBMVs) of Helicoverpa armigera. Homologous-competition assays with (125)I-Cry2Ab demonstrated that this toxin binds with high affinity to binding sites in H. armigera and Helicoverpa zea midgut. Heterologous-competition assays showed a common binding site for three toxins belonging to the Cry2A family (Cry2Aa, Cry2Ab, and Cry2Ae), which is not shared by Cry1Ac. Estimation of K(d) (dissociation constant) values revealed that Cry2Ab had around 35-fold less affinity than Cry1Ac for BBMV binding sites in both insect species. Only minor differences were found regarding R(t) (concentration of binding sites) values. This study questions previous interpretations from other authors performing binding assays with Cry2A toxins and establishes the basis for the mode of action of Cry2A toxins.

  13. Specific Binding of Bacillus thuringiensis Cry2A Insecticidal Proteins to a Common Site in the Midgut of Helicoverpa Species▿

    PubMed Central

    Hernández-Rodríguez, Carmen Sara; Van Vliet, Adri; Bautsoens, Nadine; Van Rie, Jeroen; Ferré, Juan

    2008-01-01

    For a long time, it has been assumed that the mode of action of Cry2A toxins was unique and different from that of other three-domain Cry toxins due to their apparent nonspecific and unsaturable binding to an unlimited number of receptors. However, based on the homology of the tertiary structure among three-domain Cry toxins, similar modes of action for all of them are expected. To confirm this hypothesis, binding assays were carried out with 125I-labeled Cry2Ab. Saturation assays showed that Cry2Ab binds in a specific and saturable manner to brush border membrane vesicles (BBMVs) of Helicoverpa armigera. Homologous-competition assays with 125I-Cry2Ab demonstrated that this toxin binds with high affinity to binding sites in H. armigera and Helicoverpa zea midgut. Heterologous-competition assays showed a common binding site for three toxins belonging to the Cry2A family (Cry2Aa, Cry2Ab, and Cry2Ae), which is not shared by Cry1Ac. Estimation of Kd (dissociation constant) values revealed that Cry2Ab had around 35-fold less affinity than Cry1Ac for BBMV binding sites in both insect species. Only minor differences were found regarding Rt (concentration of binding sites) values. This study questions previous interpretations from other authors performing binding assays with Cry2A toxins and establishes the basis for the mode of action of Cry2A toxins. PMID:18931285

  14. Inheritance of transgenes in transgenic Bt lines resistance to Helicoerpa armigera in upland cotton.

    PubMed

    Zhang, Baolong; Guo, Wangzhen; Zhang, Tianzhen

    2013-01-01

    Six transgenic Bt cotton cultivars (lines) including GKsu12, GK19, MR1, GK5, 109B, and SGK1 are highly resistant to bollworm from the seedling to boll-setting stages in bioassays with detached cotton leaves, though there are differences in resistant level and Bt toxin content in these transgenic cottons. Genetics analysis reveals that the resistance to Helicoverpa armigera in these six transgenic Bt cotton cultivars (lines) are controlled by one pair of dominant genes. Allelic tests further demonstrate some populations are in Mendel segregation for two nonallelic genes, i.e., the inserted Bt gene in GKsu12 is nonallelic to that of SGK1, GK5, 109B, and GK19 and Bt genes in GK19 and SGK1 are likely inserted in the same or in close proximity (genetically closely linked), while some F(2) produce abnormal segregation patterns, with a segregation of resistance to Helicoerpa armigera vary between 15:1 and 3:1, though their Bt segregation fit into 15:1 by PCR analysis, suggesting Bt gene silence in these populations. Two genes silence may occur in these populations due to the homologous sequence by crossing since the silenced individuals accounted for 1/16 of the F(2) populations for allelic test. To those silenced populations, one of their parents all showed high resistance to bollworm.

  15. Specific olfactory neurons and glomeruli are associated to differences in behavioral responses to pheromone components between two Helicoverpa species

    PubMed Central

    Wu, Han; Xu, Meng; Hou, Chao; Huang, Ling-Qiao; Dong, Jun-Feng; Wang, Chen-Zhu

    2015-01-01

    Sex pheromone communication of moths helps to understand the mechanisms underlying reproductive isolation and speciation. Helicoverpa armigera and Helicoverpa assulta use (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-9-hexadecenal (Z9-16:Ald) as pheromone components in reversed ratios, 97:3 and 5:95, respectively. H. armigera also produces trace amount of (Z)-9-tetradecenal (Z9-14:Ald) in the sex pheromone gland, but H. assulta does not. Wind tunnel studies revealed that the addition of small amounts (0.3%) of Z9-14:Ald to the main pheromone blend of H. armigera increased the males' attraction, but at higher doses (1%, 10%) the same compound acted as an inhibitor. In H. assulta, Z9-14:Ald reduced male attraction when presented as 1% to the pheromone blend, but was ineffective at lower concentrations (0.3%). Three types (A–C) of sensilla trichodea in antennae were identified by single sensillum recording, responding to Z11-16:Ald, Z9-14:Ald, and both Z9-16:Ald and Z9-14:Ald, respectively. Calcium imaging in the antennal lobes (ALs) revealed that the input information of the three chemicals was transmitted to three units of the macroglomerular complex (MGC) in ALs in both species: a large glomerulus for the major pheromone components, a small one for the minor pheromone components, and a third one for the behavioral antagonists. The type A and C neurons tuned to Z11-16:Ald and Z9-16:Ald had a reversed target in the MGC between the two species. In H. armigera, low doses (1, 10 μg) of Z9-14:Ald dominantly activated the glomerulus which processes the minor pheromone component, while a higher dose (100 μg) also evoked an equal activity in the antagonistic glomerulus. In H. assulta, instead, Z9-14:Ald always strongly activated the antagonistic glomerulus. These results suggest that Z9-14:Ald plays different roles in the sexual communication of two Helicoverpa species through activation of functionally different olfactory pathways. PMID:26300751

  16. History and Current Status of Development and Use of Viral Insecticides in China

    PubMed Central

    Sun, Xiulian

    2015-01-01

    The use of insect viruses as biological control agents started in the early 1960s in China. To date, more than 32 viruses have been used to control insect pests in agriculture, forestry, pastures, and domestic gardens in China. In 2014, 57 products from 11 viruses were authorized as commercial viral insecticides by the Ministry of Agriculture of China. Approximately 1600 tons of viral insecticidal formulations have been produced annually in recent years, accounting for about 0.2% of the total insecticide output of China. The development and use of Helicoverpa armigera nucleopolyhedrovirus, Mamestra brassicae nucleopolyhedrovirus, Spodoptera litura nucleopolyhedrovirus, and Periplaneta fuliginosa densovirus are discussed as case studies. Additionally, some baculoviruses have been genetically modified to improve their killing rate, infectivity, and ultraviolet resistance. In this context, the biosafety assessment of a genetically modified Helicoverpa armigera nucleopolyhedrovirus is discussed. PMID:25609304

  17. History and current status of development and use of viral insecticides in China.

    PubMed

    Sun, Xiulian

    2015-01-20

    The use of insect viruses as biological control agents started in the early 1960s in China. To date, more than 32 viruses have been used to control insect pests in agriculture, forestry, pastures, and domestic gardens in China. In 2014, 57 products from 11 viruses were authorized as commercial viral insecticides by the Ministry of Agriculture of China. Approximately 1600 tons of viral insecticidal formulations have been produced annually in recent years, accounting for about 0.2% of the total insecticide output of China. The development and use of Helicoverpa armigera nucleopolyhedrovirus, Mamestra brassicae nucleopolyhedrovirus, Spodoptera litura nucleopolyhedrovirus, and Periplaneta fuliginosa densovirus are discussed as case studies. Additionally, some baculoviruses have been genetically modified to improve their killing rate, infectivity, and ultraviolet resistance. In this context, the biosafety assessment of a genetically modified Helicoverpa armigera nucleopolyhedrovirus is discussed.

  18. Trade-offs of host use between generalist and specialist Helicoverpa sibling species: adult oviposition and larval performance.

    PubMed

    Liu, Zhudong; Scheirs, Jan; Heckel, David G

    2012-02-01

    Much attention has been paid to the question of the relative importance of female behaviour versus larval feeding capacities in determining the host range of herbivorous insects. Host-use trade-offs displayed by generalist and specialist sister species of the genus Helicoverpa were evaluated to examine the relationship between maternal choice and offspring performance. The prediction of optimal oviposition theory, that females will choose to lay eggs on plants on which their offspring perform best as larvae, was tested by measuring oviposition preference and larval performance of Helicoverpa armigera and H. assulta on tobacco, sunflower, and hot pepper. These two measures were more highly correlated in the specialist H. assulta. Both species exhibited the same oviposition preference ranking: tobacco > sunflower > hot pepper. H. armigera larvae preferred sunflower, followed by tobacco and hot pepper; while H. assulta larvae preferred tobacco to sunflower and hot pepper, consistent with their mothers' oviposition preference. Duration of the total period from egg to adult emergence for each species was significantly shorter on the host plant preferred by the larvae. H. assulta had shorter larval duration and higher relative growth rate than H. armigera on tobacco and hot pepper, and vice versa for sunflower, indicating species differences in host utilization. Thus, while only the specialist H. assulta displayed the predicted optimal oviposition pattern, females of both species show the least preference for the plant on which their offspring perform worst. Selection for optimal oviposition may be stronger on the specialist, which has fewer choices and lower lifetime fecundity than the generalist.

  19. Gut Transcription in Helicoverpa zea is Dynamically Altered in Response to Baculovirus Infection.

    PubMed

    Noland, Jeffrey E; Breitenbach, Jonathan E; Popham, Holly J R; Hum-Musser, Sue M; Vogel, Heiko; Musser, Richard O

    2013-09-23

    The Helicoverpa zea transcriptome was analyzed 24 h after H. zea larvae fed on artificial diet laced with Helicoverpa zea single nucleopolyhedrovirus (HzSNPV). Significant differential regulation of 1,139 putative genes (p < 0.05 T-test with Benjamini and Hochberg False Discovery Rate) was detected in the gut epithelial tissue; where 63% of these genes were down-regulated and 37% of genes were up-regulated compared to the mock-infected control. Genes that play important roles in digestive physiology were noted as being generally down-regulated. Among these were aminopeptidases, trypsin-like serine proteases, lipases, esterases and serine proteases. Genes related to the immune response reacted in a complex nature having peptidoglycan binding and viral antigen recognition proteins and antiviral pathway systems down-regulated, whereas antimicrobial peptides and prophenoloxidase were up-regulated. In general, detoxification genes, specifically cytochrome P450 and glutathione S-transferase were down-regulated as a result of infection. This report offers the first comparative transcriptomic study of H. zea compared to HzSNPV infected H. zea and provides further groundwork that will lead to a larger understanding of transcriptional perturbations associated with viral infection and the host response to the viral insult in what is likely the most heavily infected tissue in the insect.

  20. A tale of two trapping methods: Helicoverpa spp. (Lepidoptera, Noctuidae) in pheromone and light traps in Australian cotton production systems.

    PubMed

    Baker, G H; Tann, C R; Fitt, G P

    2011-02-01

    Pheromone and light traps have often been used in ecological studies of two major noctuid pests of agriculture in Australia, Helicoverpa armigera and H. punctigera. However, results from these two methods have rarely been compared directly. We set pheromone and light traps adjacent to or amongst cotton and various other crops for 10-11 years in the Namoi Valley, in northern New South Wales, Australia. Catches in pheromone traps suggested a major peak in (male) numbers of H. punctigera in early spring, with relatively few moths caught later in the summer cropping season. In contrast, (male) H. armigera were most abundant in late summer. Similar trends were apparent for catches of both male and female H. armigera in light traps, but both sexes of H. punctigera were mostly caught in mid-summer. For both species, males were more commonly caught than females. These catch patterns differed from some previous reports. At least three generations of both species were apparent in the catches. There was some evidence that the abundance of later generations could be predicted from the size of earlier generations; but, unlike previous authors, we found no positive relationships between local winter rainfall and subsequent catches of moths, nor did we find persuasive evidence of correlations between autumn and winter rainfall in central Australia and the abundance of subsequent 1st generation H. punctigera moths. Female H. punctigera were consistently more mature (gravid) and more frequently mated than those of H. armigera. Overall, our results highlight the variability in trap catches of these two species and question the use of trap catches and weather as predictors of future abundance in cropping regions.

  1. Genetic variation and virulence of Autographa californica multiple nucleopolyhedrovirus and Trichoplusia ni single nucleopolyhedrovirus isolates

    USDA-ARS?s Scientific Manuscript database

    To determine the genetic diversity within the baculovirus species Autographa calfornica multiple nucleopolyhedrovirus (AcMNPV; Baculoviridae: Alphabaculovirus), a PCR-based method was used to identify and classify baculoviruses found in virus samples from the lepidopteran host species A. californi...

  2. Binding Site Alteration Is Responsible for Field-Isolated Resistance to Bacillus thuringiensis Cry2A Insecticidal Proteins in Two Helicoverpa Species

    PubMed Central

    Caccia, Silvia; Hernández-Rodríguez, Carmen Sara; Mahon, Rod J.; Downes, Sharon; James, William; Bautsoens, Nadine; Van Rie, Jeroen; Ferré, Juan

    2010-01-01

    Background Evolution of resistance by target pests is the main threat to the long-term efficacy of crops expressing Bacillus thuringiensis (Bt) insecticidal proteins. Cry2 proteins play a pivotal role in current Bt spray formulations and transgenic crops and they complement Cry1A proteins because of their different mode of action. Their presence is critical in the control of those lepidopteran species, such as Helicoverpa spp., which are not highly susceptible to Cry1A proteins. In Australia, a transgenic variety of cotton expressing Cry1Ac and Cry2Ab (Bollgard II) comprises at least 80% of the total cotton area. Prior to the widespread adoption of Bollgard II, the frequency of alleles conferring resistance to Cry2Ab in field populations of Helicoverpa armigera and Helicoverpa punctigera was significantly higher than anticipated. Colonies established from survivors of F2 screens against Cry2Ab are highly resistant to this toxin, but susceptible to Cry1Ac. Methodology/Principal Findings Bioassays performed with surface-treated artificial diet on neonates of H. armigera and H. punctigera showed that Cry2Ab resistant insects were cross-resistant to Cry2Ae while susceptible to Cry1Ab. Binding analyses with 125I-labeled Cry2Ab were performed with brush border membrane vesicles from midguts of Cry2Ab susceptible and resistant insects. The results of the binding analyses correlated with bioassay data and demonstrated that resistant insects exhibited greatly reduced binding of Cry2Ab toxin to midgut receptors, whereas no change in 125I-labeled-Cry1Ac binding was detected. As previously demonstrated for H. armigera, Cry2Ab binding sites in H. punctigera were shown to be shared by Cry2Ae, which explains why an alteration of the shared binding site would lead to cross-resistance between the two Cry2A toxins. Conclusion/Significance This is the first time that a mechanism of resistance to the Cry2 class of insecticidal proteins has been reported. Because we found the same

  3. Analysis of the Choristoneura fumiferana nucleopolyhedrovirus genome.

    PubMed

    de Jong, Jondavid G; Lauzon, Hilary A M; Dominy, Cliff; Poloumienko, Arkadi; Carstens, Eric B; Arif, Basil M; Krell, Peter J

    2005-04-01

    The double-stranded DNA genome of Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) was sequenced and analysed in the context of other group I nucleopolyhedroviruses (NPVs). The genome consists of 129,593 bp with a G + C content of 50.1 mol%. A total of 146 open reading frames (ORFs) of greater than 150 bp, and with no or minimal overlap were identified. In addition, five homologous regions were identified containing 7-10 repeats of a 36 bp imperfect palindromic core. Comparison with other completely sequenced baculovirus genomes revealed that 139 of the CfMNPV ORFs have homologues in at least one other baculovirus and seven ORFs are unique to CfMNPV. Of the 117 CfMNPV ORFs common to all group I NPVs, 12 are exclusive to group I NPVs. Overall, CfMNPV is most similar to Orgyia pseudotsugata MNPV based on gene content, arrangement and overall amino acid identity. Unlike other group I baculoviruses, however, CfMNPV encodes a viral enhancing factor (vef) and has two copies of p26.

  4. Complete Mitochondrial Genome of Helicoverpa zea (Lepidoptera: Noctuidae) and Expression Profiles of Mitochondrial-Encoded Genes in Early and Late Embryos.

    PubMed

    Perera, Omaththage P; Walsh, Thomas K; Luttrell, Randall G

    2016-01-01

    The mitochondrial genome (mitogenome) of the bollworm, Helicoverpa zea (Boddie), was assembled using paired-end nucleotide sequence reads generated with a next-generation sequencing platform. Assembly resulted in a mitogenome of 15,348 bp with greater than 17,000-fold average coverage. Organization of the H. zea mitogenome (gene order and orientation) was identical to other known lepidopteran mitogenome sequences. Compared with Helicoverpa armigera (Hübner) mitogenome, there were a few differences in the lengths of gaps between genes, but the lengths of nucleotide overlaps were essentially conserved between the two species. Nucleotide composition of the H. zea mitochondrial genome was very similar to those of the related species H. armigera and Helicoverpa punctigera Wallengren. Mapping of RNA-Seq reads obtained from 2-h eggs and 48-h embryos to protein coding genes (PCG) revealed that all H. zea PCGs were processed as single mature gene transcripts except for the bicistronic atp8 + atp6 transcript. A tRNA-like sequence predicted to form a hammer-head-like secondary structure that may play a role in transcription start and mitogenome replication was identified within the control region of the H. zea mitogenome. Similar structures were also found within the control regions of several other lepidopteran species. Expression analysis revealed significant differences in levels of expression of PCGs within each developmental stage, but the pattern of variation was similar in both developmental stages analyzed in this study. Mapping of RNA-Seq reads to PCG transcripts also identified transcription termination and polyadenylation sites that differed from the sites described in other lepidopteran species. Published by Oxford University Press on behalf of the Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the United States.

  5. Complete Mitochondrial Genome of Helicoverpa zea (Lepidoptera: Noctuidae) and Expression Profiles of Mitochondrial-Encoded Genes in Early and Late Embryos

    PubMed Central

    Perera, Omaththage P.; Walsh, Thomas K.; Luttrell, Randall G.

    2016-01-01

    The mitochondrial genome (mitogenome) of the bollworm, Helicoverpa zea (Boddie), was assembled using paired-end nucleotide sequence reads generated with a next-generation sequencing platform. Assembly resulted in a mitogenome of 15,348 bp with greater than 17,000-fold average coverage. Organization of the H. zea mitogenome (gene order and orientation) was identical to other known lepidopteran mitogenome sequences. Compared with Helicoverpa armigera (Hübner) mitogenome, there were a few differences in the lengths of gaps between genes, but the lengths of nucleotide overlaps were essentially conserved between the two species. Nucleotide composition of the H. zea mitochondrial genome was very similar to those of the related species H. armigera and Helicoverpa punctigera Wallengren. Mapping of RNA-Seq reads obtained from 2-h eggs and 48-h embryos to protein coding genes (PCG) revealed that all H. zea PCGs were processed as single mature gene transcripts except for the bicistronic atp8 + atp6 transcript. A tRNA-like sequence predicted to form a hammer-head-like secondary structure that may play a role in transcription start and mitogenome replication was identified within the control region of the H. zea mitogenome. Similar structures were also found within the control regions of several other lepidopteran species. Expression analysis revealed significant differences in levels of expression of PCGs within each developmental stage, but the pattern of variation was similar in both developmental stages analyzed in this study. Mapping of RNA-Seq reads to PCG transcripts also identified transcription termination and polyadenylation sites that differed from the sites described in other lepidopteran species. PMID:27126963

  6. Long-term changes in the numbers of Helicoverpa punctigera (Lepidoptera: Noctuidae) in a cotton production landscape in northern New South Wales, Australia.

    PubMed

    Baker, G H; Tann, C R

    2017-04-01

    Two noctuid moths, Helicoverpa punctigera and Helicoverpa armigera, are pests of several agricultural crops in Australia, most notably cotton. Cotton is a summer crop, grown predominantly in eastern Australia. The use of transgenic (Bt) cotton has reduced the damage caused by Helicoverpa spp., but the development of Bt resistance in these insects remains a threat. In the past, large populations of H. punctigera have built up in inland Australia, following autumn-winter rains. Moths have then migrated to the cropping regions in spring, when their inland host plants dried off. To determine if there have been any long-term changes in this pattern, pheromone traps were set for H. punctigera throughout a cropping landscape in northern New South Wales from 1992 to 2015. At least three generations of moths were caught from spring to autumn. The 1st generation (mostly spring migrants) was the most numerous. Trap captures varied between sites and decreased in time, especially for moths in the 1st generation. Nearby habitat type influenced the size of catch and there was some evidence that local weather also influenced the numbers of moths caught. There was no correlation between trap catches in the cropping region and rainfall in the inland. In addition, there was little evidence that Bt cotton has reduced the abundance of H. punctigera at landscape scale. The apparent decline in the number of presumably Bt susceptible moths arriving each spring in the cropping regions from inland habitats is of concern in relation to the management of Bt resistance.

  7. Frequency of alleles conferring resistance to the Bacillus thuringiensis toxins Cry1Ac and Cry2Ab in Australian populations of Helicoverpa punctigera (Lepidoptera: Noctuidae) from 2002 to 2006.

    PubMed

    Downes, S; Parker, T L; Mahon, R J

    2009-04-01

    Helicoverpa punctigera and Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) are important pests of field and horticultural crops in Australia. The former is endemic to the continent, whereas the latter is also distributed in Africa and Asia. Although H. armigera rapidly developed resistance to virtually every group of insecticide used against it, there is only one report of resistance to an insecticide in H. punctigera. In 1996 the Australian cotton industry adopted Ingard, which expresses the Bacillus thuringiensis (Bt) toxin gene cry1Ac. In 2004/2005, Bollgard II (which expresses Cry1Ac and Cry2Ab) replaced Ingard and has subsequently been grown on 80% of the area planted to cotton, Gossypium hirsutum L. From 2002/2003 to 2006/2007, F2 screens were used to detect resistance to Cry1Ac or Cry2Ab. We detected no alleles conferring resistance to Cry1Ac; the frequency was < 0.0005 (n = 2,180 alleles), with a 95% credibility interval between 0 and 0.0014. However, during the same period, we detected alleles that confer resistance to Cry2Ab at a frequency of 0.0018 (n = 2,192 alleles), with a 95% credibility interval between 0.0005 and 0.0040. For both toxins, the experiment-wise detection probability was 94%, i.e., if there actually was a resistance allele in any tested lines, we would have detected it 94% of the time. The first isolation of Cry2Ab resistance in H. punctigera was before the widespread deployment of Bollgard II. This finding supports our published notion for H. armigera that alleles conferring resistance to Cry2Ab may be present at detectable frequencies in populations before selection by transgenic crops.

  8. Genetic diversity among isolates of Autographa californica multiple nucleopolyhedrovirus

    USDA-ARS?s Scientific Manuscript database

    Our knowledge of genetic variation at the nucleotide sequence level of Autographa californica multiple nucleopolyhedrovirus (AcMNPV; Baculoviridae: Alphabaculovirus) derives from complete genome sequences of the C6 clonal isolate of AcMNPV and the R1 and CL3 clonal isolates of AcMNPV variants Rachip...

  9. Characterization of the Lymantria dispar nucleopolyhedrovirus 25K FP gene

    Treesearch

    David S. Bischoff; James M. Slavicek

    1996-01-01

    The Lymantria dispar nucleopolyhedrovirus (LdMNPV) gene encoding the 25K FP protein has been cloned and sequenced. The 25KFP gene codes for a 217 amino acid protein with a predicted molecular mass of 24870 Da. Expression of the 25K FP protein in a rabbit reticulocyte system generated a 27 kDa protein, in close agreement with the...

  10. Both Lymantria dispar nucleopolyhedrovirus enhancin genes contribute to viral potency

    Treesearch

    Holly J.R. Popham; David S. Bischoff; James M. Slavicek

    2001-01-01

    Enhancins are a group of proteins first identified in granuloviruses (GV) that have the ability to enhance nuclear polyhedrosis virus potency. We had previously identified an enhancin gene (E1) in the Lymantria dispar multinucleocapsid nucleopolyhedrovirus (LdMNPV) (D.S. Bischoff and J.M. Slavicek, J. Virol. 71:...

  11. Relative potencies of gypsy moth nucleopolyhedrovirus genotypes isolated from Gypchek

    Treesearch

    J.D. Podgwaite; R.T. Zerillo; J.M. Slavicek; N. Hayes-Plazolles

    2011-01-01

    Gypchek is a gypsy moth (Lymantria dispar L.) - specific biopesticide whose primary use is for treating areas where environmental concerns outweigh the use of broad-spectrum pesticides for gypsy moth management. Gypchek is a lyophilized powder produced from larvae that have been infected with the gypsy moth nucleopolyhedrovirus (LdMNPV). The product...

  12. Interactions between nucleopolyhedroviruses and polydnaviruses in larval lepidoptera. Chapter 8

    Treesearch

    Vincent D' Amico; James. Slavicek

    2012-01-01

    The field dynamics of some insect populations are strongly influenced by two types of insect viruses: the nucleopolyhedroviruses (NPVs) and the polydnaviruses (PDVs). Although greatly different in origin and mode of infection, both viruses produce considerable mortality directly and indirectly in the field, and have evolved reproductive strategies that use the same...

  13. Characteristics of resistance to Bacillus thuringiensis toxin Cry2Ab in a strain of Helicoverpa punctigera (Lepidoptera: Noctuidae) isolated from a field population.

    PubMed

    Downes, S; Parker, T L; Mahon, R J

    2010-12-01

    In 1996, the Australian cotton industry adopted Ingard that expresses the Bacillus thuringiensis (Bt) toxin gene cry1Ac and was planted at a cap of 30%. In 2004-2005, Bollgard II, which expresses cry1Ac and cry2Ab, replaced Ingard in Australia, and subsequently has made up >80% of the area planted to cotton, Gossypium hirsutum L. The Australian target species Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) are innately moderately tolerant to Bt toxins, but the absence of a history of insecticide resistance indicates that the latter species is less likely to develop resistance to Bt cotton. From 2002-2003 to 2006-2007, F2 screens were deployed to detect resistance to CrylAc or Cry2Ab in natural populations of H. punctigera. Alleles that conferred an advantage against CrylAc were not detected, but those that conferred resistance to Cry2Ab were present at a frequency of 0.0018 (n = 2,192 alleles). Importantly, the first isolation of Cry2Ab resistance in H. punctigera occurred before significant opportunities to develop resistance in response to Bollgard II. We established a colony (designated Hp4-13) consisting of homozygous resistant individuals and examined their characteristics through comparison with individuals from a Bt-susceptible laboratory colony. Through specific crosses and bioassays, we established that the resistance present in Hp4-13 is due to a single autosomal gene. The resistance is fully recessive. Homozygotes are able to survive a dose of Cry2Ab toxin that is 15 times the reported concentration in field grown Bollgard II in Australia (500 microg/ml) and are fully susceptible to Cry1Ac and to the Bt product DiPel. These characteristics are the same as those described for the first Cry2Ab resistant strain of H. armigera isolated from a field population in Australia.

  14. Expression in Antennae and Reproductive Organs Suggests a Dual Role of an Odorant-Binding Protein in Two Sibling Helicoverpa Species

    PubMed Central

    Sun, Ya-Lan; Huang, Ling-Qiao; Pelosi, Paolo; Wang, Chen-Zhu

    2012-01-01

    Odorant-binding proteins (OBPs) mediate both perception and release of semiochemicals in insects. These proteins are the ideal targets for understanding the olfactory code of insects as well as for interfering with their communication system in order to control pest species. The two sibling Lepidopteran species Helicoverpa armigera and H. assulta are two major agricultural pests. As part of our aim to characterize the OBP repertoire of these two species, here we focus our attention on a member of this family, OBP10, particularly interesting for its expression pattern. The protein is specifically expressed in the antennae of both sexes, being absent from other sensory organs. However, it is highly abundant in seminal fluid, is transferred to females during mating and is eventually found on the surface of fertilised eggs. Among the several different volatile compounds present in reproductive organs, OBP10 binds 1-dodecene, a compound reported as an insect repellent. These results have been verified in both H. armigera and H. assulta with no apparent differences between the two species. The recombinant OBP10 binds, besides 1-dodecene, some linear alcohols and several aromatic compounds. The structural similarity of OBP10 with OBP1 of the mosquito Culex quinquefasciatus, a protein reported to bind an oviposition pheromone, and its affinity with 1-dodecene suggest that OBP10 could be a carrier for oviposition deterrents, favouring spreading of the eggs in these species where cannibalism is active among larvae. PMID:22291900

  15. Genomic sequence analysis of the Illinois strain of the Agrotis ipsilon multiple nucleopolyhedrovirus

    USDA-ARS?s Scientific Manuscript database

    The Agrotis ipsilon multiple nucleopolyhedrovirus (AgipMNPV) is a group II nucleopolyhedrovirus (NPV) from the black cutworm, A. ipsilon, with potential as a biopesticide to control infestations of cutworm larvae. The genome of the Illinois strain of AgipMNPV was completely sequenced. The AgipMNPV...

  16. Autographa californica multiple nucleopolyhedrovirus and Choristoneura fumiferana multiple nucleopolyhedrovirus v-cath genes are expressed as pre-proenzymes.

    PubMed

    Hodgson, Jeffrey J; Arif, Basil M; Krell, Peter J

    2009-04-01

    Intracellular processing and trafficking of the baculovirus v-cath expressed cathepsin (V-CATH), which lacks canonical targeting signals, are poorly understood. The cathepsins of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), Choristoneura fumiferana multiple nucleopolyhedrovirus (CfMNPV) and most other alphabaculovirus group I nucleopolyhedroviruses have well-conserved N-termini containing overlapping chymotrypsin-cleavage (Y(11)) and myristoylation (G(12)) motifs, which are suggestive of proteolytic signal-peptide cleavage to generate proV-CATH and subsequent acylation. To determine proteolytic N-terminal processing of V-CATH, haemagglutinin epitope-coding tags were fused to the 5' and/or 3' ends of AcMNPV and CfMNPV v-cath. Immunoblot analysis suggested that a small N-terminal peptide is cleaved for both viruses, indicating that v-cath is expressed as a pre-proenzyme. The two viral homologues undergo similar proteolytic processing, but have different glycosylation or other post-translational modifications. An AcMNPV V-CATH-DsRED fusion protein co-localized to the endoplasmic reticulum with an HDEL motif-containing green fluorescent protein. Based on these findings, pre-proV-CATH processing and trafficking mechanisms are postulated.

  17. Molecular characterization of Agrotis segetum nucleopolyhedrovirus from Poland.

    PubMed

    Jakubowska, Agata; van Oers, Monique M; Ziemnicka, Jadwiga; Lipa, Jerzy J; Vlak, Just M

    2005-09-01

    The turnip moth, Agrotis segetum (Lepidoptera, Noctuidae), is an important pest insect in Europe, Asia, and Africa. We have genetically characterized and classified a nucleopolyhedrovirus isolated from A. segetum larvae in Poland (AgseNPV-P). The restriction pattern of AgseNPV-P was distinct from an isolate from England/France (AgseNPV-UK and AgseNPV-F). Sequence analysis of three conserved baculovirus genes, polyhedrin, lef-8 and pif-2, revealed that AgseNPV-P differs substantially from the already described NPVs isolated from A. segetum and possibly represents a new NPV species. Phylogenetic analysis placed AgseNPV-P among group II NPVs and showed the closest relationship to Agrotis ipsilon (Agip) NPV and Spodoptera exigua (Se) MNPV.

  18. Cold hardiness of Helicoverpa zea (Lepidoptera: Noctuidae) pupae

    Treesearch

    A.C. Morey; W.D. Hutchison; R.C. Venette; E.C. Burkness

    2012-01-01

    An insect's cold hardiness affects its potential to overwinter and outbreak in different geographic regions. In this study, we characterized the response of Helicoverpa zea (Boddie) pupae to low temperatures by using controlled laboratory measurements of supercooling point (SCP), lower lethal temperature (LT50), and lower...

  19. Geographic isolates of Lymantria dispar multiple nucleopolyhedrovirus: Genome sequence analysis and pathogenicity against European and Asian gypsy moth strains

    USDA-ARS?s Scientific Manuscript database

    Geographic isolates of Lymantria dispar multiple nucleopolyhedrovirus: Genome sequence analysis and pathogenicity against European and Asian gypsy moth strains. To evaluate the genetic diversity of Lymantria dispar nucleopolyhedrovirus (LdMNPV) at the genomic level, the genomes of three isolates of...

  20. A Lysine at the C-Terminus of an Odorant-Binding Protein is Involved in Binding Aldehyde Pheromone Components in Two Helicoverpa Species

    PubMed Central

    Sun, Ya-Lan; Huang, Ling-Qiao; Pelosi, Paolo; Wang, Chen-Zhu

    2013-01-01

    Odorant-binding proteins (OBPs) are soluble proteins, whose role in olfaction of insects is being recognized as more and more important. We have cloned, expressed and purified an OBP (HarmOBP7) from the antennae of the moth Helicoverpa armigera. Western blot experiments indicate specific expression of this protein in the antennae of adults. HarmOBP7 binds both pheromone components Z-11-hexadecenal and Z-9-hexadecenal with good affinity. We have also performed a series of binding experiments with linear aldehydes, alcohols and esters, as well as with other compounds and found a requirement of medium size for best affinity. The affinity of OBP7, as well as that of a mutant lacking the last 6 residues does not substantially decrease in acidic conditions, but increases at basic pH values with no significant differences between wild-type and mutant. Binding to both pheromone components, instead, is negatively affected by the lack of the C-terminus. A second mutant, where one of the three lysine residues in the C-terminus (Lys123) was replaced by methionine showed reduced affinity to both pheromone components, as well as to their analogues, thus indicating that Lys123 is involved in binding these compounds, likely forming hydrogen bonds with the functional groups of the ligands. PMID:23372826

  1. Adaptive management of pest resistance by Helicoverpa species (Noctuidae) in Australia to the Cry2Ab Bt toxin in Bollgard II® cotton

    PubMed Central

    Downes, Sharon; Mahon, Rodney J; Rossiter, Louise; Kauter, Greg; Leven, Tracey; Fitt, Gary; Baker, Geoff

    2010-01-01

    In Australia, monitoring Helicoverpa species for resistance to the Cry2Ab toxin in second generation Bacillus thuringiensis (Bt) cotton has precisely fulfilled its intended function: to warn of increases in resistance frequencies that may lead to field failures of the technology. Prior to the widespread adoption of two-gene Bt cotton, the frequency of Cry2Ab resistance alleles was at least 0.001 in H. armigera and H. punctigera. In the 5 years hence, there has been a significant and apparently exponential increase in the frequency of alleles conferring Cry2Ab resistance in field populations of H. punctigera. Herein we review the history of deploying and managing resistance to Bt cotton in Australia, outline the characteristics of the isolated resistance that likely impact on resistance evolution, and use a simple model to predict likely imminent resistance frequencies. We then discuss potential strategies to mitigate further increases in resistance frequencies, until the release of a third generation product. These include mandating larger structured refuges, applying insecticide to crops late in the season, and restricting the area of Bollgard II® cotton. The area planted to Bt-crops is anticipated to continue to rise worldwide; therefore the strategies being considered in Australia are likely to relate to other situations. PMID:25567948

  2. Choristoneura fumiferana nucleopolyhedrovirus encodes a functional 3'-5' exonuclease.

    PubMed

    Yang, Dan-Hui; de Jong, Jondavid G; Makhmoudova, Amina; Arif, Basil M; Krell, Peter J

    2004-12-01

    The Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) encodes an ORF homologous to type III 3'-5' exonucleases. The CfMNPV v-trex ORF was cloned into the Bac-to-Bac baculovirus expression-vector system, expressed in insect Sf21 cells with an N-terminal His tag and purified to homogeneity by using Ni-NTA affinity chromatography. Biochemical characterization of the purified V-TREX confirmed that this viral protein is a functional 3'-5' exonuclease that cleaves oligonucleotides from the 3' end in a stepwise, distributive manner, suggesting a role in proofreading during viral DNA replication and DNA repair. Enhanced degradation of a 5'-digoxigenin- or 5'-(32)P-labelled oligo(dT)(30) substrate was observed at increasing incubation times or increased amounts of V-TREX. The 3'-excision activity of V-TREX was maximal at alkaline pH (9.5) in the presence of 5 mM MgCl(2), 2 mM dithiothreitol and 0.1 mg BSA ml(-1).

  3. Gender-mediated differences in vertical transmission of a nucleopolyhedrovirus.

    PubMed

    Virto, Cristina; Zárate, Carlos A; López-Ferber, Miguel; Murillo, Rosa; Caballero, Primitivo; Williams, Trevor

    2013-01-01

    With the development of sensitive molecular techniques for detection of low levels of asymptomatic pathogens, it becoming clear that vertical transmission is a common feature of some insect pathogenic viruses, and likely to be essential to virus survival when opportunities for horizontal transmission are unfavorable. Vertical transmission of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) is common in natural populations of S. exigua. To assess whether gender affected transgenerational virus transmission, four mating group treatments were performed using healthy and sublethally infected insects: i) healthy males (H♂) × healthy females (H♀); ii) infected males (I♂) × healthy females (H♀); iii) healthy males (H♂) × infected females (I♀) and iv) infected males (I♂) × infected females (I♀). Experimental adults and their offspring were analyzed by qPCR to determine the prevalence of infection. Both males and females were able to transmit the infection to the next generation, although female-mediated transmission resulted in a higher prevalence of infected offspring. Male-mediated venereal transmission was half as efficient as maternally-mediated transmission. Egg surface decontamination studies indicated that the main route of transmission is likely transovarial rather than transovum. Both male and female offspring were infected by their parents in similar proportions. Incorporating vertically-transmitted genotypes into virus-based insecticides could provide moderate levels of transgenerational pest control, thereby extending the periods between bioinsecticide applications.

  4. Gender-Mediated Differences in Vertical Transmission of a Nucleopolyhedrovirus

    PubMed Central

    Virto, Cristina; Zárate, Carlos A.; López-Ferber, Miguel; Murillo, Rosa; Caballero, Primitivo; Williams, Trevor

    2013-01-01

    With the development of sensitive molecular techniques for detection of low levels of asymptomatic pathogens, it becoming clear that vertical transmission is a common feature of some insect pathogenic viruses, and likely to be essential to virus survival when opportunities for horizontal transmission are unfavorable. Vertical transmission of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) is common in natural populations of S. exigua. To assess whether gender affected transgenerational virus transmission, four mating group treatments were performed using healthy and sublethally infected insects: i) healthy males (H♂)×healthy females (H♀); ii) infected males (I♂)×healthy females (H♀); iii) healthy males (H♂)×infected females (I♀) and iv) infected males (I♂)×infected females (I♀). Experimental adults and their offspring were analyzed by qPCR to determine the prevalence of infection. Both males and females were able to transmit the infection to the next generation, although female-mediated transmission resulted in a higher prevalence of infected offspring. Male-mediated venereal transmission was half as efficient as maternally-mediated transmission. Egg surface decontamination studies indicated that the main route of transmission is likely transovarial rather than transovum. Both male and female offspring were infected by their parents in similar proportions. Incorporating vertically-transmitted genotypes into virus-based insecticides could provide moderate levels of transgenerational pest control, thereby extending the periods between bioinsecticide applications. PMID:23940671

  5. Mamestra configurata nucleopolyhedrovirus-A transcriptome from infected host midgut.

    PubMed

    Donly, B Cameron; Theilmann, David A; Hegedus, Dwayne D; Baldwin, Douglas; Erlandson, Martin A

    2014-02-01

    Infection of an insect by a baculovirus occurs in two distinct phases, an initial infection of host midgut by occlusion-derived virions (ODVs) and subsequent systemic infection of other tissues by budded virions (BV). A vast majority of investigations of the infection process have been restricted to cell culture studies using BV that emulate the systemic phase of infection. This is one of the first studies to investigate baculovirus gene expression in ODV infected midgut cells. We have focused on the critical first phase of in vivo infection by Mamestra configurata nucleopolyhedrovirus-A in M. configurata larvae, using qPCR and RNAseq mass sequencing to measure virus gene expression in midgut cells. The earliest genes detected by each method had significant overlap, including known early genes as well as genes unique to MacoNPV-A and genes of unknown function. The RNAseq data also revealed a large range of expression levels across all ORFs, which could not be measured using qPCR. This dataset provides a first whole genome transcriptomic analysis of viral genes required for virus infection in vivo and will provide the basis for functionally analyzing specific genes that may be critical elements in baculovirus midgut infectivity and host range.

  6. Determination and analysis of the genome sequence of Spodoptera littoralis multiple nucleopolyhedrovirus

    USDA-ARS?s Scientific Manuscript database

    The Spodoptera littoralis multiple nucleopolyhedrovirus (SpliMNPV), a pathogen of the Egyptian cotton leaf worm Spodoptera littoralis, was subjected to sequencing of its entire DNA genome and bioassay analysis comparing its virulence to that of other baculoviruses. The annotated SpliMNPV genome of...

  7. Potency of nucleopolyhedrovirus genotypes for European and Asian gypsy moth (Lepidoptera: Lymantriidae)

    Treesearch

    J.D. Podgwaite; V.V. Martemyanov; J.M. Slavicek; S.A. Bakhavalov; S.V. Pavlushin; N. Hayes-Plazolles; R.T. Zerillo

    2013-01-01

    Gypchek is a gypsy nucleopolyhedrovirus (LdMNPV) product used for management of European gypsy moth (Lymantria dispar dispar L.) in the Unlted States, primarily in areas where the use of broad-spectrum pesticides is not appropriate. Similar LdMNPV products are used in Russia for control of a flighted-female strain of Asian gypsy moth (...

  8. Classification, genetic variation and pathogenicity of Lymantria dispar nucleopolyhedrovirus isolates from Asia, Europe, and North America

    USDA-ARS?s Scientific Manuscript database

    Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) has been formulated and applied to control outbreaks of the gypsy moth, L. dispar. To classify and determine the degree of genetic variation among isolates of L. dispar NPVs from different parts of the range of the gypsy moth, partial sequence...

  9. The Lymantria dispar nucleopolyhedrovirus enhancins are components of occlusion-derived virus

    Treesearch

    James M. Slavicek; Holly J.R. Popham

    2005-01-01

    Enhancins are metalloproteinases, first identified in granuloviruses, that can enhance nucleopolyhedrovirus (NIPV) potency. We had previously identified two enhamin genes (El and E2) in the Lymantria dispar multinucleocapsid NPV (LdMNPV) and showed that both were functional. For this study, we have extended our analysis of LdMNPV...

  10. Classification, genetic variation and pathogenicity of Lymantria dispar nucleopolyhedrovirus isolates from Asia, Europe, and North America

    Treesearch

    Robert L. Harrison; Melody A. Keena; Daniel L. Rowley

    2014-01-01

    Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) has been formulated and applied to control outbreaks of the gypsy moth, L. dispar. To classify and determine the degree of genetic variation among isolates of L. dispar NPVs from different parts of the range of the gypsy moth, partial sequences of the

  11. Complete Genome Sequences of Five Chrysodeixis chalcites Nucleopolyhedrovirus Genotypes from a Canary Islands Isolate

    PubMed Central

    Bernal, Alexandra; Williams, Trevor; Muñoz, Delia; Caballero, Primitivo

    2013-01-01

    The Chrysodeixis chalcites single nucleopolyhedrovirus (ChchSNPV) infects and kills C. chalcites larvae, an important pest of banana crops in the Canary Islands. Five genotypes present in the most prevalent and widespread isolate in the Canary Islands were sequenced, providing genetic data relevant to the genotypic and phenotypic diversity of this virus. PMID:24158555

  12. Comparative infectivity of homologous and heterologous nucleopolyhedroviruses against beet armyworm larvae

    USDA-ARS?s Scientific Manuscript database

    Homologous and heterologous nucleopolyhedroviruses (NPVs) were assayed to determine the most effective NPV against beet armyworm larvae, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae)(SeMNPV). Included were three isolates from S. exigua, one isolate each from S. littoralis Boisduval, S. litura...

  13. Fruit and vegetable extracts as radiation protectants for the beet armyworm nucleopolyhedrovirus

    USDA-ARS?s Scientific Manuscript database

    Extracts from 37 fruits and vegetables were tested as ultraviolet (UV) protectants for the nucleopolyhedrovirus (SeMNPV) of the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Only one extract (black currant) provided almost complete protection following ultraviolet B/ultraviole...

  14. Genetic and biological variation among nucleopolyhedrovirus isolates from spodoptera frugiperda (lepidotpera: noctuidae)

    USDA-ARS?s Scientific Manuscript database

    A PCR-based method was used to identify and distinguish among 40 uncharacterized nucleopolyhedrovirus (NPV) isolates from the moth Spodoptera frugiperda that were part of an insect virus collection. Phylogenetic analysis was carried out with sequences amplified from two strongly conserved loci (pol...

  15. Complete Genome Sequences of Six Chrysodeixis includens Nucleopolyhedrovirus Isolates from Brazil and Guatemala

    PubMed Central

    Craveiro, Saluana R.; Santos, Luis Arthur V. M.; Togawa, Roberto C.; Inglis, Peter W.; Grynberg, Priscila; Ribeiro, Zilda Maria A.; Ribeiro, Bergmann M.

    2016-01-01

    The baculovirus, Chrysodeixis (formerly Pseudoplusia) includens nucleopolyhedrovirus (ChinNPV), is a new Alphabaculovirus pathogenic to Chrysodeixis includens. Here, we report the complete genome sequences of six ChinNPV isolates. The availability of these genome sequences will provide information on ChinNPV molecular genetics, promoting understanding of its pathogenicity, diversity, and evolution. PMID:27932639

  16. Effects of spinosad and neem on the efficacy of a nucleopolyhedrovirus on pickleworm larvae

    USDA-ARS?s Scientific Manuscript database

    A neem formulation (Neemix® 4.5) and spinosad (SpinTor® 2SC) were tested for their effects when mixed with the multicapsid nucleopolyhedrovirus virus (AgMNPV) from the velvetbean caterpillar, Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), for control of pickleworm larvae, Diaphania nitidalis...

  17. Proteomics of the Autographa californica nucleopolyhedrovirus budded virions.

    PubMed

    Wang, Ranran; Deng, Fei; Hou, Dianhai; Zhao, Yong; Guo, Lin; Wang, Hualin; Hu, Zhihong

    2010-07-01

    Baculoviruses produce two progeny phenotypes during their replication cycles. The occlusion-derived virus (ODV) is responsible for initiating primary infection in the larval midgut, and the budded virus (BV) phenotype is responsible for the secondary infection. The proteomics of several baculovirus ODVs have been revealed, but so far, no extensive analysis of BV-associated proteins has been conducted. In this study, the protein composition of the BV of Autographa californica nucleopolyhedrovirus (AcMNPV), the type species of baculoviruses, was analyzed by various mass spectrometry (MS) techniques, including liquid chromatography-triple quadrupole linear ion trap (LC-Qtrap), liquid chromatography-quadrupole time of flight (LC-Q-TOF), and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF). SDS-PAGE and MALDI-TOF analyses showed that the three most abundant proteins of the AcMNPV BV were GP64, VP39, and P6.9. A total of 34 viral proteins associated with the AcMNPV BV were identified by the indicated methods. Thirteen of these proteins, PP31, AC58/59, AC66, IAP-2, AC73, AC74, AC114, AC124, chitinase, polyhedron envelope protein (PEP), AC132, ODV-E18, and ODV-E56, were identified for the first time to be BV-associated proteins. Western blot analyses showed that ODV-E18 and ODV-E25, which were previously thought to be ODV-specific proteins, were also present in the envelop fraction of BV. In addition, 11 cellular proteins were found to be associated with the AcMNPV BV by both LC-Qtrap and LC-Q-TOF analyses. Interestingly, seven of these proteins were also identified in other enveloped viruses, suggesting that many enveloped viruses may commonly utilize certain conserved cellular pathways.

  18. BV/ODV-E26: a palmitoylated, multifunctional structural protein of Autographa californica nucleopolyhedrovirus.

    PubMed

    Burks, Jared K; Summers, Max D; Braunagel, Sharon C

    2007-04-25

    Autographa californica nucleopolyhedrovirus Ac16 is 1 of 17 genes conserved within Type 1 nucleopolyhedroviruses. This report demonstrates that multiple isoforms of the protein encoded by Ac16, BV/ODV-E26 (E26), are present in the infected cell. One form of E26 associates with viral DNA or DNA-binding proteins, while a second form associates with intracellular membranes and this is likely due to palmitoylation. The different forms of E26 present unique epitopes that can be discriminated by antiserum produced to bacterially or virally produced antigen. A summation of the data now available on E26 suggests that it is a multifunctional protein and the functional states assume unique conformations that can be discriminated by differing antisera.

  19. The mosaic structure of the polyhedrin gene of the Autographa californica nucleopolyhedrovirus (AcMNPV).

    PubMed

    Jehle, Johannes A

    2004-08-01

    The polyhedrin (polh) gene of nucleopolyhedroviruses (NPVs) encodes for the matrix protein of the virus occlusion body and is one of the most conserved baculovirus genes. Previous analyses of different NPV genes and polh genes provided conflicting results indicating that the Autographa californica nucleopolyhedrovirus (AcMNPV) is generally a member of the so-called group I NPVs and is most closely related to Rachiplusia ou (Ro) NPV, whereas the AcMNPV polh is more similar to the polh of the group II NPVs. A comparative analysis of the AcMNPV polh and its closest neighbours within group I and group II NPV, the RoMNPV and the Thysanoplusia orichalcea (Thor) NPV, was performed using Hidden Markov Models for detecting recombination. The result provided strong evidence that the AcMNPV polh is a chimerical gene which consists of a mosaic of group I and group II NPV specific sequences.

  20. The Lymantria dispar nucleopolyhedrovirus contains the capsid-associated p24 protein gene

    Treesearch

    James M. Slavicek; Nancy Hayes-Plazolles

    2003-01-01

    During the course of investigations on a wild-type strain of Lymantria dispar multinucleocapsid nucleopolyhedrovirus (LdMNPV), a region of the viral genome was analyzed and found to contain 697 bp that is lacking in the sequenced strain (5-6) of LdMNPV (Kuzio et al., Virology 253, 17-34, 1999). The sequenced strain of LdMNPV contains a mutation in...

  1. Complete Genome Sequence of Lymantria dispar multiple nucleopolyhedrovirus Isolated in Southwestern Poland

    PubMed Central

    Krejmer-Rabalska, Martyna; Skrzecz, Iwona; Szewczyk, Boguslaw

    2016-01-01

    We report here the complete genomic sequence of Polish alphabaculovirus isolated from dead gypsy moth caterpillars. Its genome structure and G+C content indicate that it is a Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) strain which possesses the shortest sequence among eight other (similar to reference sequence AF081810) LdMNPV sequences available in GenBank; it is 159,729 bp long. PMID:28007858

  2. Biosafety of Recombinant and Wild Type Nucleopolyhedroviruses as Bioinsecticides

    PubMed Central

    Ashour, Mohamed-Bassem; Ragheb, Didair A.; El-Sheikh, El-Sayed A.; Gomaa, El-Adarosy A.; Kamita, Shizuo G.; Hammock, Bruce D.

    2007-01-01

    The entomopathogenic Autographa californica (Speyer) nucleopolyhedrovirus (AcMNPV) has been genetically modified to increase its speed of kill. The potential adverse effects of a recombinant AcMNPV (AcAaIT) as well as wild type AcMNPV and wild type Spodoptera littoralis NPV (SlNPV) were studied. Cotton plants were treated with these viruses at concentrations that were adjusted to resemble the recommended field application rate (4 × 1012 PIBs/feddan, feddan = 4,200 m2) and 3rd instar larvae of S. littoralis were allowed to feed on the contaminated plants. SDS-PAGE, ELISA, and DNA analyses were used to confirm that larvae that fed on these plants were virus-infected. Polyhedra that were purified from the infected larvae were subjected to structural protein analysis. A 32 KDa protein was found in polyhedra that were isolated from all of the viruses. Subtle differences were found in the size and abundance of ODV proteins. Antisera against polyhedral proteins isolated from AcAaIT polyhedra were raised in rabbits. The terminal bleeds from rabbits were screened against four coating antigens (i.e., polyhedral proteins from AcAaIT, AcAaIT from field-infected larvae (AcAaIT-field), AcMNPV, and SlNPV) using a two-dimensional titration method with the coated antigen format. Competitive inhibition experiments were conducted in parallel to optimize antibody and coating antigen concentrations for ELISA. The IC50 values for each combination ranged from 1.42 to 163 μg/ml. AcAaIT-derived polyhedrin gave the lowest IC50 value, followed by those of SlNPV, AcAaIT-field, and AcMNPV. The optimized ELISA system showed low cross reactivity for AcMNPV (0.87%), AcAaIT-field (1.2%), and SlNPV (4.0%). Genomic DNAs isolated from AcAaIT that were passaged in larvae of S. littoralis that were reared in the laboratory or field did not show any detectable differences. Albino rats (male and female) that were treated with AcAaIT, AcMNPV or SlNPV (either orally or by intraperitoneal injection at

  3. Biosafety of recombinant and wild type nucleopolyhedroviruses as bioinsecticides.

    PubMed

    Ashour, Mohamed-Bassem; Ragheb, Didair A; El-Sheikh, El-Sayed A; Gomaa, El-Adarosy A; Kamita, Shizuo G; Hammock, Bruce D

    2007-06-01

    The entomopathogenic Autographa californica (Speyer) nucleopolyhedrovirus (AcMNPV) has been genetically modified to increase its speed of kill. The potential adverse effects of a recombinant AcMNPV (AcAaIT) as well as wild type AcMNPV and wild type Spodoptera littoralis NPV (SlNPV) were studied. Cotton plants were treated with these viruses at concentrations that were adjusted to resemble the recommended field application rate (4 x 10(12) PIBs/feddan, feddan = 4,200 m2) and 3rd instar larvae of S. littoralis were allowed to feed on the contaminated plants. SDS-PAGE, ELISA, and DNA analyses were used to confirm that larvae that fed on these plants were virus-infected. Polyhedra that were purified from the infected larvae were subjected to structural protein analysis. A 32 KDa protein was found in polyhedra that were isolated from all of the viruses. Subtle differences were found in the size and abundance of ODV proteins. Antisera against polyhedral proteins isolated from AcAaIT polyhedra were raised in rabbits. The terminal bleeds from rabbits were screened against four coating antigens (i.e., polyhedral proteins from AcAaIT, AcAaIT from field-infected larvae (AcAaIT-field), AcMNPV, and SlNPV) using a two-dimensional titration method with the coated antigen format. Competitive inhibition experiments were conducted in parallel to optimize antibody and coating antigen concentrations for ELISA. The IC50 values for each combination ranged from 1.42 to 163 microg/ml. AcAaIT-derived polyhedrin gave the lowest IC50 value, followed by those of SlNPV, AcAaIT-field, and AcMNPV. The optimized ELISA system showed low cross reactivity for AcMNPV (0.87%), AcAaIT-field (1.2%), and SlNPV (4.0%). Genomic DNAs isolated from AcAaIT that were passaged in larvae of S. littoralis that were reared in the laboratory or field did not show any detectable differences. Albino rats (male and female) that were treated with AcAaIT, AcMNPV or SlNPV (either orally or by intraperitoneal injection at

  4. Comparative activity of Choristoneura fumiferana nucleopolyhedrovirus propagated in different hosts.

    PubMed

    Ebling, Peter M

    2004-07-01

    The biological activity of the Ireland strain of Choristoneura fumiferana (Clem) nucleopolyhedrovirus (CfMNPV) propagated in different hosts was determined to provide the basis upon which genetically modified CfMNPV, or other naturally occurring isolates, should be compared. Occlusion bodies (OB) derived from CF-203 cells were significantly larger and more pathogenic than those propagated in vivo when tested against the fifth larval instar of C fumiferana (Clem) and C occidentalis Freeman. The dose-responses (LD50 and LD95, expressed as occlusion bodies per larva) of C fumiferana larvae to in vitro-propagated OBs were 274 and 5785, respectively. The values of LD50 and LD95 to C occidentalis larvae were 19 and 118, respectively. There were no significant differences in pathogenicity or size when OBs propagated in C fumiferana larvae were tested against either insect species, nor were there significant differences for OBs propagated in C occidentalis larvae. The LD50 and LD95 of in vivo-produced OBs to C fumiferana were 925 and 61988, respectively. The LD50 and LD95 to C occidentalis were 50 and 453, respectively. OBs propagated in vitro had a mean volume of 13.13 microm3, whereas those propagated in vivo ranged from 0.84 to 1.41 microm3. The median survival time-responses (ST50) of fifth-instar C fumiferana or C occidentalis larvae to OBs propagated in vivo were not significantly different from those propagated in vitro at the dosage levels tested. Values of ST50 of C fumiferana larvae to in vitro- and in vivo-produced OBs at dosages causing less than 50% mortality rangedfrom 9.6 to 9.8 days post-inoculation (dpi), whereas a LD95 dose resulted in ST50 values ranging from 7.3 to 7.7 days. ST50 values of C occidentalis larvae at dosages causing less than 50% mortality ranged from 9.8 to 10.2 dpi, whereas a LD95 dose resulted in ST50 values ranging from 9.5 to 9.8 dpi. The median feeding cessation time-response (FT50) of fifth-instar C fumiferana larvae to OBs

  5. The expression of three opsin genes from the compound eye of Helicoverpa armigera (Lepidoptera: Noctuidae) is regulated by a circadian clock, light conditions and nutritional status.

    PubMed

    Yan, Shuo; Zhu, Jialin; Zhu, Weilong; Zhang, Xinfang; Li, Zhen; Liu, Xiaoxia; Zhang, Qingwen

    2014-01-01

    Visual genes may become inactive in species that inhabit poor light environments, and the function and regulation of opsin components in nocturnal moths are interesting topics. In this study, we cloned the ultraviolet (UV), blue (BL) and long-wavelength-sensitive (LW) opsin genes from the compound eye of the cotton bollworm and then measured their mRNA levels using quantitative real-time PCR. The mRNA levels fluctuated over a daily cycle, which might be an adaptation of a nocturnal lifestyle, and were dependent on a circadian clock. Cycling of opsin mRNA levels was disturbed by constant light or constant darkness, and the UV opsin gene was up-regulated after light exposure. Furthermore, the opsin genes tended to be down-regulated upon starvation. Thus, this study illustrates that opsin gene expression is determined by multiple endogenous and exogenous factors and is adapted to the need for nocturnal vision, suggesting that color vision may play an important role in the sensory ecology of nocturnal moths.

  6. Structure and transcription of the Helicoverpa armigera densovirus (HaDV2) genome and its expression strategy in LD652 cells.

    PubMed

    Xu, Pengjun; Graham, Robert I; Wilson, Kenneth; Wu, Kongming

    2017-02-07

    Densoviruses (DVs) are highly pathogenic to their hosts. However, we previously reported a mutualistic DV (HaDV2). Very little was known about the characteristics of this virus, so herein we undertook a series of experiments to explore the molecular biology of HaDV2 further. Phylogenetic analysis showed that HaDV2 was similar to members of the genus Iteradensovirus. However, compared to current members of the genus Iteradensovirus, the sequence identity of HaDV2 is less than 44% at the nucleotide-level, and lower than 36, 28 and 19% at the amino-acid-level of VP, NS1 and NS2 proteins, respectively. Moreover, NS1 and NS2 proteins from HaDV2 were smaller than those from other iteradensoviruses due to their shorter N-terminal sequences. Two transcripts of about 2.2 kb coding for the NS proteins and the VP proteins were identified by Northern Blot and RACE analysis. Using specific anti-NS1 and anti-NS2 antibodies, Western Blot analysis revealed a 78 kDa and a 48 kDa protein, respectively. Finally, the localization of both NS1 and NS2 proteins within the cell nucleus was determined by using Green Fluorescent Protein (GFP) labelling. The genome organization, terminal hairpin structure, transcription and expression strategies as well as the mutualistic relationship with its host, suggested that HaDV2 was a novel member of the genus Iteradensovirus within the subfamily Densovirinae.

  7. The Expression of Three Opsin Genes from the Compound Eye of Helicoverpa armigera (Lepidoptera: Noctuidae) Is Regulated by a Circadian Clock, Light Conditions and Nutritional Status

    PubMed Central

    Yan, Shuo; Zhu, Jialin; Zhu, Weilong; Zhang, Xinfang; Li, Zhen; Liu, Xiaoxia; Zhang, Qingwen

    2014-01-01

    Visual genes may become inactive in species that inhabit poor light environments, and the function and regulation of opsin components in nocturnal moths are interesting topics. In this study, we cloned the ultraviolet (UV), blue (BL) and long-wavelength-sensitive (LW) opsin genes from the compound eye of the cotton bollworm and then measured their mRNA levels using quantitative real-time PCR. The mRNA levels fluctuated over a daily cycle, which might be an adaptation of a nocturnal lifestyle, and were dependent on a circadian clock. Cycling of opsin mRNA levels was disturbed by constant light or constant darkness, and the UV opsin gene was up-regulated after light exposure. Furthermore, the opsin genes tended to be down-regulated upon starvation. Thus, this study illustrates that opsin gene expression is determined by multiple endogenous and exogenous factors and is adapted to the need for nocturnal vision, suggesting that color vision may play an important role in the sensory ecology of nocturnal moths. PMID:25353953

  8. Effects of different Brush Border Membrane Vesicle isolation protocols on proteomic analysis of Cry1Ac binding proteins from the midgut of Helicoverpa armigera

    USDA-ARS?s Scientific Manuscript database

    The brush border membrane vesicles (BBMV) isolated from midgut cells of insect have been widely used for studying of the binding receptors and action mode of Cry proteins produced by Bacillus thuringiensis (Bt). There are several methods for isolating insect BBMV used in one-dimension electrophoresi...

  9. Development of transgenic CryIA(c) + GNA cotton plants via pollen tube pathway method confers resistance to Helicoverpa armigera and Aphis gossypii Glover.

    PubMed

    Liu, Zhi; Zhu, Zhen; Zhang, Tianzhen

    2013-01-01

    Elite cotton cultivar Sumian16 was transformed with p7RPSBK-mGNA-NPTII containing Bt (CryIA(c)), Galanthus nivalis agglutinin (GNA) resistance genes and selectable marker NptII gene via the pollen tube pathway method and two fertile transgenic Bt + GNA plants were obtained in the present study. The integration and expression of the Bt and GNA genes were confirmed by molecular biology techniques and insect bioassays. Insect bioassays showed that the transformed plants were highly toxic to bollworm larvae as well as obviously retarding development of aphid populations. PCR analyses and identification of resistance to Kanamycin and bollworm showed that the resistance to bollworm for the two transgenic plants was dominantly inherited in a Mendelian manner and the two resistance genes and selectable marker co-segregated from primary transformed parents to the first self-fertilized progeny plants.

  10. A microRNA encoded by Autographa californica nucleopolyhedrovirus regulates expression of viral gene ODV-E25.

    PubMed

    Zhu, Mengxiao; Wang, Jinwen; Deng, Riqiang; Xiong, Peiwen; Liang, Hai; Wang, Xunzhang

    2013-12-01

    Baculovirus-encoded microRNAs (miRNAs) have been described in Bombyx mori nucleopolyhedrovirus; however, most of their functions remain unclear. Here we report the identification and characterization of an miRNA encoded by Autographa californica nucleopolyhedrovirus. The identified miRNA, AcMNPV-miR-1, perfectly matched a segment in the coding sequence of the viral gene ODV-E25 and downregulated ODV-E25 mRNA expression, which likely resulted in a reduction of infectious budded virions and accelerated the formation of occlusion-derived virions.

  11. Gene organization and sequencing of the Choristoneura fumiferana defective nucleopolyhedrovirus genome.

    PubMed

    Lauzon, Hilary A M; Jamieson, Peter B; Krell, Peter J; Arif, Basil M

    2005-04-01

    Two distinct nucleopolyhedrovirus species of the eastern spruce budworm, Choristoneura fumiferana, exist in a symbiont-like relationship. C. fumiferana defective nucleopolyhedrovirus (CfDEFNPV) only infects C. fumiferana larvae per os in the presence of C. fumiferana nucleopolyhedrovirus Ireland strain (CfMNPV), but is infective when injected into the haemolymph. CfDEFNPV synergizes CfMNPV in per os infections and CfMNPV is always the predominant progeny. This study was undertaken to report the genomic makeup and organization of CfDEFNPV in an attempt to identify its defect and understand its synergistic role. The genome was mapped, sequenced, characterized and compared to other baculoviruses. The CfDEFNPV genome was 131,160 nt long with 149 putative open reading frames (ORFs) and a G + C content of 45.8 mol%. Homologues of all 62 conserved lepidopteran baculovirus genes were found including those implicated in per os infectivity, p74, per os infectivity factor (pif) and pif-2. Although no obvious deletions were observed to explain the defect, two ORFs, Cfdef79 and Cfdef99 (inhibitor of apoptosis-4), contained potential deletions. Cfdef50 (late expression factor-10)/Cfdef51 (vp1054) and Cfdef76/Cfdef77 (telokin-like protein) had large overlaps and a potential homologue to ac105/he65 was split. Four baculovirus repeat ORFs were present, as were two unique genes, but no enhancins were identified. CfDEFNPV contained 13 homologous regions, each with one to five palindromes. Comparison with fully sequenced baculovirus genomes identified CfDEFNPV as a group I NPV with the closest average amino acid identity to Epiphyas postvittana NPV, followed by Orgyia pseudotsugata MNPV and CfMNPV, with its closest matches being to individual Anticarsia gemmatalis MNPV gene sequences.

  12. Entry of Bombyx mori nucleopolyhedrovirus into BmN cells by cholesterol-dependent macropinocytic endocytosis.

    PubMed

    Huang, Jinshan; Hao, Bifang; Cheng, Chen; Liang, Fei; Shen, Xingjia; Cheng, Xiaowen

    2014-10-10

    Bombyx mori nucleopolyhedrovirus (BmNPV) is a serious viral pathogen of silkworm, and no drug or specific protection against BmNPV infection is available at present time. Although functions of most BmNPV genes were depicted in recent years, knowledge on the mechanism of BmNPV entry into insect cells is still limited. Here BmNPV cell entry mechanism is investigated by different endocytic inhibitor application and subcellular analysis. Results indicated that BmNPV enters BmN cells by clathrin-independent macropinocytic endocytosis, which is mediated by cholesterol in a dose-dependent manner, and cholesterol replenishment rescued the BmNPV infection partially.

  13. The autographa californica multiple nucleopolyhedrovirus ODV-E56 envelope protein is required for oral infectivity and can be functionally substituted by rachiplusia ou multiple nucleopolyhedrovirus ODV-E56

    USDA-ARS?s Scientific Manuscript database

    The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) odv-e56 gene encodes an occlusion-derived virus (ODV)-specific envelope protein, ODV-E56. In a previous analysis, the odv-e56 gene was found to be under positive selection pressure, suggesting that it may be a determinant of viral ho...

  14. Helicoverpa zea and Bt cotton in the United States.

    PubMed

    Luttrell, Randall G; Jackson, Ryan E

    2012-01-01

    Helicoverpa zea (Boddie), the bollworm or corn earworm, is the most important lepidopteran pest of Bt cotton in the United States. Corn is the preferred host, but the insect feeds on most flowering crops and wild host plants. As a cotton pest, bollworm has been closely linked to the insecticide-resistance prone Heliothis virescens (F.), tobacco budworm. Immature stages of the two species are difficult to separate in field environments. Tobacco budworm is very susceptible to most Bt toxins, and Bt cotton is considered to be "high dose." Bollworm is less susceptible to Bt toxins, and Bt cotton is not "high dose" for this pest. Bt cotton is routinely sprayed with traditional insecticides for bollworm control. Assays of bollworm field populations for susceptibility to Bt toxins expressed in Bt cotton have produced variable results since pre-deployment of Bt cottons in 1988 and 1992. Analyses of assay response trends have been used by others to suggest that field resistance has evolved to Bt toxins in bollworm, but disagreement exists on definitions of field resistance and confidence of variable assay results to project changes in susceptibility of field populations. Given historical variability in bollworm response to Bt toxins, erratic field control requiring supplemental insecticides since early field testing of Bt cottons, and dramatic increases in corn acreage in cotton growing areas of the Southern US, continued vigilance and concern for resistance evolution are warranted.

  15. Concentration- and time-response characteristics of plaque isolates of Agrotis ipsilon multiple nucleopolyhedrovirus derived from a field isolate

    USDA-ARS?s Scientific Manuscript database

    Plaque isolates derived from the Illinois field isolate of Agrotis ipsilon multiple nucleopolyhedrovirus are distinguished by the presence or absence of a small deletion in the baculovirus egt (ecdysteroid UDP-glucosyltransferase) coding sequence. Dose-response and time-response bioassays were perf...

  16. A soluble form of P74 can act as a per os infectivity factor to the autographa californica multiple nucleopolyhedrovirus

    USDA-ARS?s Scientific Manuscript database

    The baculovirus occlusion-derived virion (ODV) is required to spread virus infection among insect hosts via the per os route. The Autographa californica Multicapsid Nucleopolyhedrovirus (AcMNPV) P74 protein is an ODV envelope protein that is essential for ODVs to be infectious. P74 is anchored in ...

  17. Geographic isolates of Lymantria dispar multiple nucleopolyhedrovirus: Genome sequence analysis and pathogenicity against European and Asian gypsy moth strains

    Treesearch

    Harrison Robert L.; Daniel L. Rowley; Melody A. Keena

    2016-01-01

    Isolates of the baculovirus species Lymantria dispar multiple nucleopolyhedrovirus have been formulated and applied to suppress outbreaks of the gypsy moth, L. dispar. To evaluate the genetic diversity in this species at the genomic level, the genomes of three isolates from Massachusetts, USA (LdMNPV-Aba624), Spain (LdMNPV-3054...

  18. A comparison of the adaptations of strains of Lymantria dispar multiple nucleopolyhedrovirus to hosts from spatially isolated populations

    Treesearch

    V.V. Martemyanov; J.D. Podgwaite; I.A. Belousova; S.V. Pavlushin; J.M. Slavicek; O.A. Baturina; M.R. Kabilov; A.V. Ilyinykh

    2017-01-01

    The adaptation of pathogens to either their hosts or to environmental conditions is the focus of many current ecological studies. In this work we compared the ability of six spatially-distant Lymantria dispar (gypsy moth) multiple nucleopolyhedrovirus (LdMNPV) strains (three from eastern North America and three from central Asia) to induce acute...

  19. Genetic variation and biological activity of isolates of lymantria dispar multiple nucleopolyhedrovirus from north america, europe, and asia

    USDA-ARS?s Scientific Manuscript database

    Little is known about genetic variation of Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV; Baculoviridae: Alphabaculovirus) at the nucleotide sequence level. To obtain a more comprehensive view of genetic diversity among isolates of LdMNPV, partial sequences of the lef-8 gene were generated...

  20. Genomic analysis of five Lymantria dispar nucleopolyhedrovirus isolates and biological activity against different host strains of Lymantria dispar

    USDA-ARS?s Scientific Manuscript database

    To evaluate genetic diversity of Lymantria dispar nucleopolyhedrovirus (LdMNPV) at the genomic level, five isolates of LdMNPV from North America, Europe, and Asia were selected for complete genome sequence determination and analysis. These isolates consist of LdMNPV-2161 from Korea; LdMNPV-3029, a ...

  1. Field Evaluation of a Kudzu/Cottonseed Oil Formulation on the Persistence of the Beet Armyworm Nucleopolyhedrovirus

    USDA-ARS?s Scientific Manuscript database

    A plant extract (kudzu) was tested as a UV protectant for SeMNPV, with and without the addition of an oil/emulsifier (cottonseed oil/lecithin) formulation. Aqueous and oil emulsion formulations of the beet armyworm, Spodoptera exigua (Hübner), nucleopolyhedrovirus SeMNPV were applied to collards an...

  2. Involvement of Bombyx mori nucleopolyhedrovirus ORF41 (Bm41) in BV production and ODV envelopment

    SciTech Connect

    Tian Caihong; Zhao Jinfang; Xu Yipeng; Xue Jian; Zhang Baoqin; Cui Yingjun; Zhang Minjuan; Bao Yanyuan; Zhang Chuanxi

    2009-04-25

    Bombyx mori nucleopolyhedrovirus (BmNPV) ORF41 (Bm41), homologous to Ac52, is a gene present in most lepidopteran nucleopolyhedroviruses. Bm41 transcripts and encoded protein in BmNPV-infected cells can be detected from 3 and 6 h post-infection, respectively. Immunoassays have shown that Bm41 is not a viral structural protein and is detected in both the nuclei and cytoplasm of infected cells. A Bm41-disrupted virus (vBm{sup De}) and a repaired virus (vBm{sup Re}) were generated to investigate the function of Bm41. The results showed that Bm41 was essential for viral replication, and the disruption of Bm41 resulted in a much lower viral titer. Transmission electron microscopy revealed that disruption of Bm41 affected normal nucleocapsid envelopment and polyhedra formation in the nucleus. The disruption of Bm41 might severely affect odv-ec27 and polyhedrin expression. The disrupted virus reduced BmNPV infectivity in an LD{sub 50} bioassay and took 18-23 h longer to kill larvae than wild-type virus in an LT{sub 50} bioassay.

  3. ac18 is not essential for the propagation of Autographa californica multiple nucleopolyhedrovirus

    SciTech Connect

    Wang Yanjie; Wu Wenbi; Li Zhaofei; Yuan Meijin; Feng Guozhong; Yu Qian; Yang Kai Pang Yi

    2007-10-10

    orf18 (ac18) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved gene in lepidopteran nucleopolyhedroviruses, but its function remains unknown. In this study, an ac18 knockout AcMNPV bacmid was generated to determine the role of ac18 in baculovirus life cycle. After transfection of Sf-9 cells, the ac18-null mutant showed similar infection pattern to the parent virus and the ac18 repair virus with respect to the production of infectious budded virus, occlusion bodies, or the formation of nucleocapsids as visualized by electron microscopy. The deletion mutant did not reduce AcMNPV infectivity for Trichoplusia ni in LD{sub 50} bioassay; however, it did take 24 h longer for deleted mutant to kill T. ni larvae than wild-type virus in LT{sub 50} bioassay. Our results demonstrate that ac18 is not essential for viral propagation both in vitro and in vivo, but it may play a role in efficient virus infection in T. ni larvae.

  4. The first record of nucleopolyhedrovirus isolated from the gypsy moth, Lymantria dispar (Lepidoptera, Lymantriidae) in Turkey.

    PubMed

    Yaman, Mustafa; Bekircan, Cağrı; Radek, Renate; Linde, Andreas

    2012-01-01

    The gypsy moth, Lymantria dispar L. (Lepidoptera, Lymantriidae) is a common pest of forests and fruit trees throughout the world. This insect is also a major serious pest in Turkey. Nowadays L. dispar can be managed by biological control methods especially, using entomopathogenic viruses. The aim of this study is to characterize entomopathogenic viruses and is the first record of nucleopolyhedrovirus isolated from the gypsy moth in Turkey. PIBs obtained from infected larvae were measured and photographed using an Olympus BX51 microscope with a DP-25 digital camera and a DP2-BSW Soft Imaging System and examined with a Philips 208 electron microscope (TEM). The virus had the typical characteristics of nucleopolyhedroviruses. The dimension of the polyhedral inclusion bodies (PIBs) was 2.03±0.25 µm. PIBs varied in size from 1.65 to 2.21 µm and were usually polygonal in shape. Virions in PIBs contained 1 to 8 nucleocapsids per virion. The size of the viral particles was 366.67±54.72 (312-500) x 42.95±6.12 (30-47) nm. The isolation and characterization of a pure isolate of Lymantria dispar multinucleopolyhedrovirus (LdMNPV-TR) from Turkey is presented for the first time.

  5. Involvement of Bombyx mori nucleopolyhedrovirus ORF41 (Bm41) in BV production and ODV envelopment.

    PubMed

    Tian, Cai-Hong; Zhao, Jin-Fang; Xu, Yi-Peng; Xue, Jian; Zhang, Bao-Qin; Cui, Ying-Jun; Zhang, Min-Juan; Bao, Yan-Yuan; Zhang, Chuan-Xi

    2009-04-25

    Bombyx mori nucleopolyhedrovirus (BmNPV) ORF41 (Bm41), homologous to Ac52, is a gene present in most lepidopteran nucleopolyhedroviruses. Bm41 transcripts and encoded protein in BmNPV-infected cells can be detected from 3 and 6 h post-infection, respectively. Immunoassays have shown that Bm41 is not a viral structural protein and is detected in both the nuclei and cytoplasm of infected cells. A Bm41-disrupted virus (vBm(De)) and a repaired virus (vBm(Re)) were generated to investigate the function of Bm41. The results showed that Bm41 was essential for viral replication, and the disruption of Bm41 resulted in a much lower viral titer. Transmission electron microscopy revealed that disruption of Bm41 affected normal nucleocapsid envelopment and polyhedra formation in the nucleus. The disruption of Bm41 might severely affect odv-ec27 and polyhedrin expression. The disrupted virus reduced BmNPV infectivity in an LD(50) bioassay and took 18-23 h longer to kill larvae than wild-type virus in an LT(50) bioassay.

  6. Molecular cloning and functional characterization of the diapause hormone receptor in the corn earworm Helicoverpa zea

    USDA-ARS?s Scientific Manuscript database

    The diapause hormone (DH) in the heliothine moth has shown its activity in termination of pupal diapause, while the orthology in the silkworm is known to induce embryonic diapause. In the current study, we cloned the diapause hormone receptor from the corn earworm Helicoverpa zea (HzDHr) and tested ...

  7. Effect of Hexaflumuron on feeding response and reproduction of bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae)

    USDA-ARS?s Scientific Manuscript database

    Hexaflumuron (Consult® 100 EC, Dow AgroSciences) is an insect growth regulator that inhibits chitin synthesis. The efficacy of hexaflumuron mixed with 2.5 M sucrose (ppm) was evaluated in the laboratory against bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) for toxicity, proboscis exten...

  8. Molecular identification and expression analysis of a diapause hormone receptor in the corn earworm, Helicoverpa zea

    USDA-ARS?s Scientific Manuscript database

    Diapause hormone (DH) is an insect neuropeptide that is highly effective in terminating the overwintering pupal diapause in members of the Helicoverpa/Heliothis complex of agricultural pests, thus DH and related compounds have promise as tools for pest management. To augment our development of effe...

  9. Peripheral genetic structure of Helicoverpa zea indicates asymmetrical panmixia.

    PubMed

    Seymour, Mathew; Perera, Omaththage P; Fescemyer, Howard W; Jackson, Ryan E; Fleischer, Shelby J; Abel, Craig A

    2016-05-01

    Seasonal climatic shifts create peripheral habitats that alternate between habitable and uninhabitable for migratory species. Such dynamic peripheral habitats are potential sites where migratory species could evolve high genetic diversity resulting from convergence of immigrants from multiple regionally distant areas. Migrant populations of Helicoverpa zea (Boddie) captured during two different seasons were assessed for genetic structure using microsatellite markers and for host plant type using stable carbon isotope analysis. Individuals (N = 568) were genotyped and divided into 13 putative populations based on collection site and time. Fixation indices (F-statistics), analysis of molecular variance (AMOVA), and discriminant analysis of principal components (DAPC) were used to examine within and among population genetic variation. Mean number of alleles per locus was 10.25 (± 3.2 SD), and allelic richness ranged from 2.38 to 5.13 (± 3.2 SD). The observed and expected heterozygosity ranged from 0.07 to 0.48 and 0.08 to 0.62, respectively. Low F ST (0.01 to 0.02) and high F IS (0.08 to 0.33) values suggest captured migrants originated from breeding populations with different allele frequencies. We postulate that high genetic diversity within migrant populations and low genetic differentiation among migrant populations of H. zea are the result of asymmetrical immigration due to the high dispersal and reproductive behavior of H. zea, which may hinder the adaptation and establishment of H. zea to peripheral habitat. These findings highlight the importance of assessing peripheral population structure in relation to ecological and evolutionary dynamics of this and other highly reproductive and dispersive species.

  10. Resistance to Bombyx mori nucleopolyhedrovirus via overexpression of an endogenous antiviral gene in transgenic silkworms.

    PubMed

    Jiang, Liang; Wang, Genhong; Cheng, Tingcai; Yang, Qiong; Jin, Shengkai; Lu, Gai; Wu, Fuquan; Xiao, Yang; Xu, Hanfu; Xia, Qingyou

    2012-07-01

    Transgenic technology is a powerful tool for improving disease-resistant species. Bmlipase-1, purified from the midgut juice of Bombyx mori, showed strong antiviral activity against B. mori nucleopolyhedrovirus (BmNPV). In an attempt to create an antiviral silkworm strain for sericulture, a transgenic vector overexpressing the Bmlipase-1 gene was constructed under the control of a baculoviral immediate early-1 (IE1) promoter. Transgenic lines were generated via embryo microinjection. The mRNA level of Bmlipase-1 in the midguts of the transgenic line was 27.3 % higher than that of the non-transgenic line. After feeding the silkworm with different amounts of BmNPV, the mortality of the transgenic line decreased to approximately 33 % compared with the non-transgenic line when the virus dose was 10(6) OB/larva. These results imply that overexpressing endogenous antiviral genes can enhance the antiviral resistance of silkworms.

  11. Identification of a Novel Apoptosis Suppressor Gene from the Baculovirus Lymantria dispar Multicapsid Nucleopolyhedrovirus

    PubMed Central

    Yamada, Hayato; Shibuya, Miyuki; Kobayashi, Michihiro; Ikeda, Motoko

    2011-01-01

    Ld652Y cells from Lymantria dispar readily undergo apoptosis upon infection with a variety of nucleopolyhedroviruses (NPVs), while L. dispar multicapsid NPV (LdMNPV) infection of Ld652Y cells results in the production of a high titer of progeny viruses. Here, we identify a novel LdMNPV apoptosis suppressor gene, apsup, which functions to suppress apoptosis induced in Ld652Y cells by infection with vAcΔp35, a p35-defective recombinant Autographa californica MNPV. apsup also suppresses apoptosis of Ld652Y cells induced by actinomycin D and UV exposure. Apsup is expressed in LdMNPV-infected Ld652Y cells late in infection, and RNA interference-mediated apsup ablation induces apoptosis of LdMNPV-infected Ld652Y cells. PMID:21411519

  12. Genome Sequence and Analysis of Buzura suppressaria Nucleopolyhedrovirus: A Group II Alphabaculovirus

    PubMed Central

    Zhu, Zheng; Yin, Feifei; Liu, Xiaoping; Hou, Dianhai; Wang, Jun; Zhang, Lei; Arif, Basil; Wang, Hualin; Deng, Fei; Hu, Zhihong

    2014-01-01

    The genome of Buzura suppressaria nucleopolyhedrovirus (BusuNPV) was sequenced by 454 pyrosequencing technology. The size of the genome is 120,420 bp with 36.8% G+C content. It contains 127 hypothetical open reading frames (ORFs) covering 90.7% of the genome and includes the 37 conserved baculovirus core genes, 84 genes found in other baculoviruses, and 6 unique ORFs. No typical baculoviral homologous repeats (hrs) were present but the genome contained a region of repeated sequences. Gene Parity Plots revealed a 28.8 kb region conserved among the alpha- and beta-baculoviruses. Overall comparisons of BusuNPV to other baculoviruses point to a distinct species in group II Alphabaculovirus. PMID:24475121

  13. Reduced expression of Autographa californica nucleopolyhedrovirus ORF34, an essential gene, enhances heterologous gene expression

    SciTech Connect

    Salem, Tamer Z.; Zhang, Fengrui; Thiem, Suzanne M.

    2013-01-20

    Autographa californica multiple nucleopolyhedrovirus ORF34 is part of a transcriptional unit that includes ORF32, encoding a viral fibroblast growth factor (FGF) and ORF33. We identified ORF34 as a candidate for deletion to improve protein expression in the baculovirus expression system based on enhanced reporter gene expression in an RNAi screen of virus genes. However, ORF34 was shown to be an essential gene. To explore ORF34 function, deletion (KO34) and rescue bacmids were constructed and characterized. Infection did not spread from primary KO34 transfected cells and supernatants from KO34 transfected cells could not infect fresh Sf21 cells whereas the supernatant from the rescue bacmids transfection could recover the infection. In addition, budded viruses were not observed in KO34 transfected cells by electron microscopy, nor were viral proteins detected from the transfection supernatants by western blots. These demonstrate that ORF34 is an essential gene with a possible role in infectious virus production.

  14. Hierarchical spatial structure of genetically variable nucleopolyhedroviruses infecting cyclic populations of western tent caterpillars.

    PubMed

    Cooper, Dawn; Cory, Jenny S; Myers, Judith H

    2003-04-01

    The cyclic population dynamics of western tent caterpillars, Malacosoma californicum pluviale, are associated with epizootics of a nucleopolyhedrovirus, McplNPV. Given the dynamic fluctuations in host abundance and levels of viral infection, host resistance and virus virulence might be expected to change during different phases of the cycle. As a first step in determining if McplNPV virulence and population structure change with host density, we used restriction fragment length polymorphism (RFLP) analysis to examine the genetic diversity of McplNPV infecting western tent caterpillar populations at different spatial scales. Thirteen dominant genetic variants were identified in 39 virus isolates (individual larvae) collected from field populations during one year of low host density, and another distinct variant was discovered among nine additional isolates in two subsequent years of declining host density. The distribution of these genetic variants was not random and indicated that the McplNPV population was structured at several spatial levels. A high proportion of the variation could be explained by family grouping, which suggested that isolates collected within a family were more likely to be the same than isolates compared among populations. Additionally, virus variants from within populations (sites) were more likely to be the same than isolates collected from tent caterpillar populations on different islands. This may indicate that there is limited mixing of virus among tent caterpillar families and populations when host population density is low. Thus there is potential for the virus to become locally adapted to western tent caterpillar populations in different sites. However, no dominant genotype was observed at any site. Whether and how selection acts on the genetically diverse nucleopolyhedrovirus populations as host density changes will be investigated over the next cycle of tent caterpillar populations.

  15. Chondroitinase from baculovirus Bombyx mori nucleopolyhedrovirus and chondroitin sulfate from silkworm Bombyx mori.

    PubMed

    Sugiura, Nobuo; Ikeda, Motoko; Shioiri, Tatsumasa; Yoshimura, Mayumi; Kobayashi, Michihiro; Watanabe, Hideto

    2013-12-01

    Chondroitin sulfate (CS) is a linear polysaccharide composed of repeating disaccharide units of glucuronic acid (GlcUA) and N-acetyl-d-galactosamine (GalNAc) with sulfate groups at various positions. Baculovirus is an insect-pathogenic virus that infects Lepidoptera larvae. Recently, we found that the occlusion-derived virus envelope protein 66 (ODV-E66) from Autographa californica nucleopolyhedrovirus (AcMNPV) exhibits chondroitin (CH)-digesting activity with distinct substrate specificity. Here, we demonstrate that the ODV-E66 protein from Bombyx mori nucleopolyhedrovirus (BmNPV) exhibits 92% homology to the amino acid sequence and 83% of the CH lyase activity of ODV-E66 from AcMNPV. ODV-E66 cleaves glycosyl bonds at nonreducing sides of disaccharide units consisting of nonsulfated and 6-O-sulfated GalNAc residues. We then investigated CS in the silkworm, Bombyx mori, which is the host of BmNPV. CS was present in insect tissues such as the midgut, peritrophic membrane, silk gland and skin. The polysaccharide consisted of a nonsulfated disaccharide unit, mono-sulfated disaccharide at Position 4 of the GalNAc residue and mono-sulfated disaccharide at Position 6 of the GalNAc residue. With regard to immunohistochemical analysis, the staining patterns of the silkworm tissues were different among anti-CS antibodies. Chondroitn sulfate that is digestible by ODV-E66 exists sufficiently in the peritrophic membrane protecting the midgut epithelium from ingested pathogens. Our results suggest that ODV-E66 facilitates the primary infection of the virus by digestion of CS in the peritrophic membrane.

  16. Multiple nucleocapsid packaging of Autographa californica nucleopolyhedrovirus accelerates the onset of systemic infection in Trichoplusia ni.

    PubMed

    Washburn, J O; Lyons, E H; Haas-Stapleton, E J; Volkman, L E

    1999-01-01

    Among the nucleopolyhedroviruses (Baculoviridae), the occlusion-derived virus (ODV), which initiates infection in host insects, may contain only a single nucleocapsid per virion (the SNPVs) or one to many nucleocapsids per virion (the MNPVs), but the significance of this difference is unclear. To gain insight into the biological relevance of these different packaging strategies, we compared pathogenesis induced by ODV fractions enriched for multiple nucleocapsids (ODV-M) or single nucleocapsids (ODV-S) of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) containing a beta-galactosidase reporter gene. In time course experiments wherein newly molted fourth-instar Trichoplusia ni were challenged with doses of ODV-S or ODV-M that yielded the same final mortality ( approximately 70%), we characterized viral foci as either being restricted to the midgut or involving tracheal cells (the secondary target tissue, indicative of systemic infection). We found that while the timing of primary infection by ODV-S and ODV-M was similar, ODV-S established significantly more primary midgut cell foci than ODV-M, but ODV-M infected tracheal cells at twice the rate of ODV-S. The more efficient establishment of tracheal infections by ODV-M decreased the probability that infections were lost by midgut cell sloughing, explaining why higher numbers of primary infections established by ODV-S within larvae were needed to achieve the same final mortality. These results showed that the multiple nucleocapsid packaging strategy of AcMNPV accelerates the onset of irreversible systemic infections and may indicate why MNPVs have wider individual host ranges than SNPVs.

  17. Classification, genetic variation and pathogenicity of Lymantria dispar nucleopolyhedrovirus isolates from Asia, Europe, and North America.

    PubMed

    Harrison, Robert L; Keena, Melody A; Rowley, Daniel L

    2014-02-01

    Lymantria dispar multiple nucleopolyhedrovirus (LdMNPV) has been formulated and applied to control outbreaks of the gypsy moth, L. dispar. To classify and determine the degree of genetic variation among isolates of L. dispar NPVs from different parts of the range of the gypsy moth, partial sequences of the lef-8, lef-9, and polh genes were determined for Lymantria spp. virus samples from host populations throughout the world. Sequence analysis confirmed that all L. dispar virus samples tested contained isolates of the species Lymantria dispar multiple nucleopolyhedrovirus (Baculoviridae: Alphabaculovirus). Phylogenetic inference based on the lef-8 sequences indicated that the LdMNPV isolates formed two groups, one consisting primarily of isolates from Asia, and one consisting primarily of isolates from Europe and North America. The complete genome sequence was determined for an isolate from the Asian group, LdMNPV-2161 (S. Korea). The LdMNPV-2161 genome was 163,138bp in length, 2092bp larger than the previously determined genome of LdMNPV isolate 5-6 (CT, USA). The two genome sequences were co-linear, with an overall nucleotide sequence identity of 97.5% and some differences in ORF content. In droplet-feeding bioassays against neonate L. dispar larvae, isolates LdMNPV-3029 (Virin-ENSh/Russia) and LdMNPV-Ab-a624 (MA, USA) killed neonate larvae with an LC50 values that were 1.8- to 3.2-fold lower than a sample of Gypchek® (CT, USA) and isolates LdMNPV-3041 (Japan) and LdMNPV-2161. This study expands our knowledge about genetic variation among LdMNPV isolates and provides novel information on the distinct groups in which these NPVs occur. Published by Elsevier Inc.

  18. Microbial control of black cutworm (Lepidoptera: Noctuidae) in turfgrass using Agrotis ipsilon multiple nucleopolyhedrovirus.

    PubMed

    Prater, Callie A; Redmond, Carl T; Barney, Walter; Bonning, Bryony C; Potter, Daniel A

    2006-08-01

    Agrotis ipsilon multiple nucleopolyhedrovirus (family Baculoviridae, genus Nucleopolyhedrovirus, AgipMNPV), a naturally occurring baculovirus, was found infecting black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), on central Kentucky golf courses. Laboratory, greenhouse, and field studies investigated the potential of AgipMNPV for managing black cutworms in turfgrass. The virus was highly active against first instars (LC50 = 73 occlusion bodies [OBs] per microl with 2-microl dose; 95% confidence intervals, 55-98). First instars that ingested a high lethal dose stopped feeding and died in 3-6 d as early second instars, whereas lethally infected fourth instars continued to feed and grow for 4-9 d until death. Sublethal doses consumed by third or fifth instars had little or no effect on subsequent developmental rate or pupal weight. Horizontal transmission of AgipMNPV in turfgrass plots was shown. Sprayed suspensions of AgipMNPV (5 x 10(8) - 6 x 10(9) OBs/m2) resulted in 75 to > 93% lethal infection of third or fourth instars in field plots of fairway-height creeping bentgrass, Agrostis stolonifera (Huds.), and on a golf course putting green collar. Virus spray residues (7 x 10(9) OBs/m2) allowed to weather on mowed and irrigated creeping bentgrass field plots significantly increased lethal infection of implanted larvae for at least 4 wk. This study, the first to evaluate a virus against a pest in turfgrass, suggests that AgipMNPV has potential as a preventive bioinsecticide targeting early instar black cutworms. Establishing a virus reservoir in the thatch and soil could suppress successive generations of that key pest on golf courses and sport fields.

  19. The genome sequence of Agrotis segetum nucleopolyhedrovirus B (AgseNPV-B) reveals a new baculovirus species within the Agrotis baculovirus complex.

    PubMed

    Wennmann, Jörg T; Gueli Alletti, Gianpiero; Jehle, Johannes A

    2015-04-01

    The genome of Agrotis segetum nucleopolyhedrovirus B (AgseNPV-B) was completely sequenced and compared with whole genome sequences of the Agrotis segetum nucleopolyhedrovirus A (AgseNPV-A) and Agrotis ipsilon nucleopolyhedrovirus (AgipNPV). The AgseNPV-B genome is 148,981 bp in length and encodes 150 putative open reading frames. AgseNPV-B contains two copies of the gene viral enhancing factor (vef), making the Agrotis nucleopolyhedroviruses and A. segetum granulovirus (AgseGV) very rich in vef in comparison to other baculoviruses. Genome alignments of AgseNPV-B, AgseNPV-A and AgipNPV showed a very high genome co-linearity interspersed with variable regions, which are considered as putative sites of genomic recombination. Phylogenetic analyses revealed that all three viruses are distinct. However, AgseNPV-B is more closely related to AgipNPV suggesting that both viruses are at an early stage of phylogenetic divergence. It is proposed that AgseNPV-B belongs to a third Alphabaculovirus species of the Agrotis baculovirus complex. The Agrotis exclamationis nucleopolyhedrovirus (AgexNPV) shared high nucleotide sequence identities with AgseNPV-B, suggesting it is actually an AgseNPV-B isolate.

  20. Downregulation of a chitin deacetylase-like protein in response to baculovirus infection and its application for improving baculovirus infectivity.

    PubMed

    Jakubowska, Agata K; Caccia, Silvia; Gordon, Karl H; Ferré, Juan; Herrero, Salvador

    2010-03-01

    Several expressed sequence tags (ESTs) with homology to chitin deacetylase-like protein (CDA) were selected from a group of Helicoverpa armigera genes whose expression changed after infection with H. armigera single nucleopolyhedrovirus (HearNPV). Some of these ESTs coded for a midgut protein containing a chitin deacetylase domain (CDAD). The expressed protein, HaCDA5a, did not show chitin deacetylase activity, but it showed a strong affinity for binding to chitin. Sequence analysis showed the lack of any chitin binding domain, described for all currently known peritrophic membrane (PM) proteins. HaCDA5a has previously been detected in the H. armigera PM. Such localization, together with its downregulation after pathogen infection, led us to hypothesize that this protein might be responsible for the homeostasis of the PM structure and that, by reduction of its expression, the insect may reduce PM permeability, decreasing the entrance of baculovirus. To test this hypothesis, we constructed a recombinant nucleopolyhedrovirus to express HaCDA5a in insect cells and tested its influence on PM permeability as well as the influence of HaCDA5a expression on the performance of the baculovirus. The experiments showed that HaCDA5a increased PM permeability, in a concentration-dependent manner. Bioassays on Spodoptera frugiperda and Spodoptera exigua larvae revealed that NPV expressing HaCDA5a was more infective than its parental virus. However, no difference in virulence was observed when the viruses were injected intrahemocoelically. These findings support the downregulation of a midgut-specific CDA-like protein as a possible mechanism used by H. armigera to reduce susceptibility to baculovirus by decreasing PM permeability.

  1. 75 FR 62484 - Importation of Shepherd's Purse With Roots From the Republic of Korea Into the United States

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-12

    ... horticola) Turnip moth (Agrotis segetum) American bollworm moth (Helicoverpa armigera) Cabbage webworm moth (Hellula undalis, Fabricius) The cabbage moth (Mamestra brassicae) Oriental leafworm moth...

  2. Evidence of recent interspecies horizontal gene transfer regarding nucleopolyhedrovirus infection of Spodoptera frugiperda.

    PubMed

    Barrera, Gloria Patricia; Belaich, Mariano Nicolás; Patarroyo, Manuel Alfonso; Villamizar, Laura Fernanda; Ghiringhelli, Pablo Daniel

    2015-11-25

    Baculoviruses are insect-associated viruses carrying large, circular double-stranded-DNA genomes with significant biotechnological applications such as biological pest control, recombinant protein production, gene delivery in mammals and as a model of DNA genome evolution. These pathogens infect insects from the orders Lepidoptera, Hymenoptera and Diptera, and have high species diversity which is expressed in their diverse biological properties including morphology, virulence or pathogenicity. Spodoptera frugiperda (Lepidoptera: Noctuidae), the fall armyworm, represents a significant pest for agriculture in America; it is a host for baculoviruses such as the Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) (Colombia strain, genotype A) having been classified as a Group II alphabaculovirus making it a very attractive target for bioinsecticidal use. Genome analysis by pyrosequencing revealed that SfMNPV ColA has 145 ORFs, 2 of which were not present in the other sequenced genotypes of the virus (SfMNPV-NicB, SfMNPV-NicG, SfMNPV-19 and SfMNPV-3AP2). An in-depth bioinformatics study showed that ORF023 and ORF024 were acquired by a recent homologous recombination process between Spodoptera frugiperda and Spodoptera litura (the Oriental leafworm moth) nucleopolyhedroviruses. Auxiliary genes are numerous in the affected locus which has a homologous region (hr3), a repetitive sequence associated with genome replication which became lost in SfColA along with 1 ORF. Besides, the mRNAs associated with two acquired genes appeared in the virus' life-cycle during the larval stage. Predictive studies concerning the theoretical proteins identified that ORF023 protein would be a phosphatase involved in DNA repair and that the ORF024 protein would be a membrane polypeptide associated with cell transport. The SfColA genome was thus revealed to be a natural recombinant virus showing evidence of recent horizontal gene transfer between different baculovirus species occurring

  3. Analysis of the Bombyx mori nucleopolyhedrovirus ie-1 promoter in insect, mammalian, plant, and bacterial cells.

    PubMed

    Fujita, Ryosuke; Ono, Chikako; Ono, Isamu; Asano, Shin-Ichiro; Bando, Hisanori

    2015-09-04

    The Bombyx mori nucleopolyhedrovirus (BmNPV) ie-1 promoter exhibits strong transcriptional activity and is used in transient foreign gene expression systems in insect cells. In a reporter assay experiment using the BmNPV ie-1 promoter, we found that it exhibited activity even in non-host mammalian BHK cells, plant BY-2 cells, and also bacterial Escherichia coli cells. An analysis using a deletion series of the BmNPV ie-1 promoter demonstrated that the core promoter region of this promoter was sufficient to display promoter activity in BHK cells, BY-2 cells, and E. coli cells, whereas upstream elements were required for higher activity in insect cells. Furthermore, we found that the BmNPV ie-1 promoter exhibited sufficient activity for a β-galactosidase assay in E. coli cells. The results obtained here suggest that the BmNPV ie-1 promoter has potential as a universal promoter for transient expression systems in insect, mammalian, plant, and bacterial cells. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Functional characterization of Autographa californica multiple nucleopolyhedrovirus gp16 (ac130)

    SciTech Connect

    Yang, Ming; Huang, Cui; Qian, Duo-Duo; Li, Lu-Lin

    2014-09-15

    To investigate the function of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) gp16, multiple gp16-knockout and repair mutants were constructed and characterized. No obvious difference in productivity of budded virus, DNA synthesis, late gene expression and morphogenesis was observed between gp16-knockout and repair viruses, but gp16 deletion resulted in six hours of lengthening in ST{sub 50} to the third instar Spodoptera exigua larvae in bioassays. GP16 was fractionated mainly in the light membrane fraction, by subcellular fractionation. A GP16-EGFP fusion protein was predominantly localized close around the nuclear membrane in infected cells, being coincident with formation of the vesicles associated with the nuclear membrane, which hosted nucleocapsids released from the nucleus. These data suggest that gp16 is not required for viral replication, but may be involved in membrane trafficking associated with the envelopment/de-envelopment of budded viruses when they cross over the nuclear membrane and pass through cytoplasm. - Highlights: • gp16 knockout and repair mutants of AcMNPV were constructed and characterized. • AcMNPV gp16 is not essential to virus replication. • Deletion of gp16 resulted in time lengthening to kill S. exigua larvae. • GP16 was localized close around the nuclear membrane of infected cells. • GP16 was fractionated in the light membrane fraction in subcellular fractionation.

  5. Autographa californica multiple nucleopolyhedrovirus ac53 plays a role in nucleocapsid assembly

    SciTech Connect

    Liu Chao; Li Zhaofei Wu Wenbi; Li Lingling; Yuan Meijin; Pan Lijing; Yang Kai Pang Yi

    2008-12-05

    Autographa californica multiple nucleopolyhedrovirus (AcMNPV) orf53 (ac53) is a highly conserved gene existing in all sequenced Lepidoptera and Hymenoptera baculoviruses, but its function remains unknown. To investigate its role in the baculovirus life cycle, an ac53 deletion virus (vAc{sup ac53KO-PH-GFP}) was generated through homologous recombination in Escherichia coli. Fluorescence and light microscopy and titration analysis revealed that vAc{sup ac53KO-PH-GFP} could not produce infectious budded virus in infected Sf9 cells. Real-time PCR demonstrated that the ac53 deletion did not affect the levels of viral DNA replication. Electron microscopy showed that many lucent tubular shells devoid of the nucleoprotein core are present in the virogenic stroma and ring zone, indicating that the ac53 knockout affected nucleocapsid assembly. With a recombinant virus expressing an Ac53-GFP fusion protein, we observed that Ac53 was distributed within the cytoplasm and nucleus at 24 h post-infection, but afterwards accumulated predominantly near the nucleus-cytoplasm boundary. These data demonstrate that ac53 is involved in nucleocapsid assembly and is an essential gene for virus production.

  6. Persistence of naturally occurring and genetically modified Choristoneura fumiferana nucleopolyhedroviruses in outdoor aquatic microcosms.

    PubMed

    Holmes, Stephen B; Fick, William E; Kreutzweiser, David P; Ebling, Peter M; England, Laura S; Trevors, Jack T

    2008-10-01

    To assess the persistence of genetically modified and naturally occurring baculoviruses in an aquatic environment, replicate (three) outdoor, aquatic microcosms were spiked with spruce budworm viruses [Ireland strain of Choristoneura fumiferana multiple nucleopolyhedrovirus (CfMNPV) and the recombinant CfMNPVegt(-)/lacZ(+)] at a rate of 1.86 x 10(10) occlusion bodies (OBs) m(-2) of surface area. The presence of virus in water samples collected at various times after inoculation was determined by PCR amplification of baculoviral DNA extracted from OBs. Although UV radiation rapidly degrades baculoviruses under natural conditions, both viruses persisted above the level of detection (>100 OBs 450 microL(-1) of natural pond water) for at least 1 year post-inoculation, with little difference between the viruses in their patterns of persistence. The present microcosm study suggests that occlusion bodies of baculoviruses can persist in the flocculent layer of natural ponds. On disturbance, OBs could re-enter the main water column and thus be available for transport to new locations. Implications for environmental risk assessment are discussed.

  7. Nuclear localization of actin requires AC102 in Autographa californica multiple nucleopolyhedrovirus-infected cells

    PubMed Central

    Ohkawa, Taro; Volkman, Loy E.

    2012-01-01

    Autographa californica multiple nucleopolyhedrovirus requires nuclear actin for progeny virus production and thereby encodes viral products that ensure actin’s translocation to and retention within the nucleus. Current evidence suggests that the ie0–ie1 gene complex along with five nuclear localization of actin (NLA) genes are sufficient for NLA in transient transfection experiments. Here we report that, during infection, only one of the five NLA genes, Ac102, was essential for NLA, and that AC102 had at least one other activity critical for budded virus (BV) production. Viral deletion mutants in the other four NLA genes were viable, with only two having replication phenotypes different from that of the wild type. Infection with AcΔpe38 revealed a delay in both BV production and NLA. Infection with AcΔ152 revealed a delay in BV production, but no corresponding delay in NLA. Infection with either AcΔpe38 or AcΔ152 resulted in slightly reduced BV titres. Deletion of Ac004 or he65 had no impact on actin translocation kinetics, timing of BV production or BV titres. These results implicate AC102 as a key player in baculovirus manipulation of actin. PMID:22592260

  8. High-level expression of canine parvovirus VP2 using Bombyx mori nucleopolyhedrovirus vector.

    PubMed

    Choi, J Y; Woo, S D; Lee, H K; Hong, H K; Je, Y H; Park, J H; Song, J Y; An, S H; Kang, S K

    2000-01-01

    For the potential use as recombinant vaccine, canine parvovirus (CPV) major capsid protein VP2 was expressed using Bombyx mori nucleopolyhedrovirus (BmNPV) vector. CPV VP2 gene was introduced into polyhedrin-based BmNPV transfer vector pBmKSK3, and recombinant virus BmK1-Parvo was prepared. When anti-CPV.VP2 monoclonal antibody was employed in immunofluorescence staining, an intense signal was observed within BmK1-Parvo-infected Bm5 cells but not within uninfected cells or cells infected with a wild-type BmNPV-K1. In hemagglutination assay, the expression level of VP2 were 3.2 x 10(3) HA units/ml from infected Bm5 cells, 2.1x 10(5) HA units/larvae from infected larval fat body, and 1.6x 10(6) HA units/ml from infected larval hemolymph. These results suggested that BmNPV vector system using B. mori larva as host could be applied to efficient mass-production of recombinant vaccines.

  9. Pathogenicity of Choristoneura fumiferana nucleopolyhedrovirus propagated in vitro at different incubation temperatures.

    PubMed

    Ebling, Peter M; Caputo, Guido F; Cook, Barbara J

    2003-01-01

    To optimize the in vitro production of Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) as a potential microbial pest control agent, the pathogenicity of occlusion bodies (OBs) produced in two cell lines at three incubation temperatures was determined by bioassay. A plaque-purified isolate of CfMNPV was amplified in permissive C. fumiferana cell lines, FPMI-CF-203 and FPMI-CF-2C1, and incubated at 22, 24, and 28 degrees C. Occlusion bodies propagated in FPMI-CF-203 cells at 28 degrees C were significantly larger (17.5 microm(3)) and more pathogenic (LD(50) = 27; LD(95) = 185, where LD(50) and LD(95) are doses required to kill 50 and 95% of the test larvae, respectively) than those produced in either of the cell lines at any of the incubation temperatures tested. Increased temperatures yielded larger OBs from both cell lines. The pathogenicity of OBs propagated in the FPMI-CF-203 cell line increased with incubation temperature, whereas that of OBs produced in FPMI-CF-2C1 cells decreased. Comparison of the pathogenicity of OBs, whether naturally occurring or genetically modified, should be standardized by cell line and incubation temperature used for propagation. Production efficiency decreased with increasing incubation temperature for each cell line. Lower incubation temperatures used for propagation, and standardization of the titer of viral inoculum, should be further investigated to determine the economic feasibility of the in vitro production of CfMNPV as a microbial pest control agent.

  10. Chitinase from Autographa californica multiple nucleopolyhedrovirus: rapid purification from Sf-9 medium and mode of action.

    PubMed

    Fukamizo, Tamo; Sato, Hirokazu; Mizuhara, Mamiko; Ohnuma, Takayuki; Gotoh, Takeshi; Hiwatashi, Kazuyuki; Takahashi, Saori

    2011-01-01

    Autographa californica multiple nucleopolyhedrovirus (AcMNPV) chitinase is involved in the final liquefaction of infected host larvae. We purified the chitinase rapidly to homogeneity from Sf-9 cells infected with AcMNPV by a simple procedure using a pepstatin-aminohexyl-Sepharose column. In past studies, a recombinant AcMNPV chitinase was found to exhibit both exo- and endo-chitinase activities by analysis using artificial substrates with a fluorescent probe. In this study, however, we obtained more accurate information on the mode of action of the chitinase by HPLC analysis of the enzymatic products using natural oligosaccharide and polysaccharide substrates. The AcMNPV chitinase hydrolyzed the second β-1,4 glycosidic linkage from the non-reducing end of the chitin oligosaccharide substrates [(GlcNAc)(n), n=4, 5, and 6], producing the β-anomer of (GlcNAc)₂. The mode of action was similar to that of Serratia marcescens chitinase A (SmChiA), the amino acid sequence of which is 60.5% homologous to that of the AcMNPV enzyme. The enzyme also hydrolyzed solid β-chitin, producing only (GlcNAc)₂. The AcMNPV chitinase processively hydrolyzes solid β-chitin in a manner similar to SmChiA. The processive mechanism of the enzyme appears to be advantageous in liquefaction of infected host larvae.

  11. Occurrence and characterization of a tetrahedral nucleopolyhedrovirus from Spilarctia obliqua (Walker).

    PubMed

    Senthil Kumar, C M; Jacob, T K; Devasahayam, S; D'Silva, Sharon; Jinsha, J; Rajna, S

    2015-11-01

    Spilarctia obliqua Walker (Lepidoptera: Arctiidae) is a polyphagous insect pest damaging pulses, oil seeds, cereals, vegetables and medicinal and aromatic plants in India. The pest also infests turmeric and ginger sporadically in Kerala. We observed an epizootic caused by a nucleopolyhedrovirus (NPV) in field populations of the insects in December 2013. The NPV was purified and characterized. The isolate was tetrahedral in shape and belonged to multicapsid NPV. The REN profile of the SpobNPV genome with Pst I, Xho I and HindIII enzymes showed a genome size of 99.1±3.9 kbp. Partialpolh, lef-8 and lef-9 gene sequences of the isolate showed a close relationship with HycuNPV and SpphNPV. Phylogram and K-2-P distances between similar isolates suggested inclusion of the present SpobNPV isolate to group I NPV. The biological activity of the isolate was tested under laboratory conditions against third instar larvae of S. obliqua and the LC50 was 4.37×10(3)OBs/ml occlusion bodies (OBs) per ml. The median survival time (ST50) was 181 h at a dose of 1×10(6)OBs/ml and 167 h at a dose of 1×10(8)OBs/ml. SpobNPV merits further field evaluation as a potential biological control agent of S. obliqua, a serious pest of many agriculturally important crops in the Oriental region. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. Sublethal dose of phoxim and Bombyx mori nucleopolyhedrovirus interact to elevate silkworm mortality.

    PubMed

    Gu, ZhiYa; Li, FanChi; Hu, JingSheng; Ding, Chao; Wang, Chaoqian; Tian, JiangHai; Xue, Bin; Xu, KaiZun; Shen, WeiDe; Li, Bing

    2017-03-01

    Silkworm (Bombyx mori) is an economically important insect. It is relatively less resistant to certain chemicals and environment exposures such as pesticides and pathogens. After pesticide exposures, the silkworms are more susceptible to microbial infections. The mechanism underlying the susceptibility might be related to immune response and oxidative stress. A sublethal dose of phoxim combined with Bombyx mori nucleopolyhedrovirus (BmNPV) elevated the silkworm mortality at 96 h. We found a higher content of H2 O2 and increased levels of genes related to oxidative stress and immune response after treatment with a sublethal dose of phoxim for 24 h or 48 h. However, such response decreased with longer pesticide treatment. Mortality increased by 44% when B. mori was exposed to combined treatment with BmNPV and phoxim rather than BmNPV alone. The level of examined immune-related and oxidative-stress-related genes significantly decreased in the combined treatment group compared with the BmNPV group. Our results indicated that, with long-term exposure to pesticides such as OPs, even at sublethal dose, the oxidative stress response and immune responses in silkworm were inhibited, which may lead to further immune impairment and accumulation of oxidative stress, resulting in susceptibility to the virus and harm to the silkworm. Our study provided insights for understanding the susceptibility to pathogen after pesticide exposures, which may promote the development of better pesticide controls to avoid significant economic losses. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  13. Characterization of bro-a gene of Spodoptera litura multicapsid nucleopolyhedrovirus.

    PubMed

    Gong, Yingxue; Li, Zhaofei; Pan, Lijing; Yu, Mei; Yang, Kai; Pang, Yi

    2004-04-01

    Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) ORF120 (designated as the bro-a gene) is a member of the unique multigene family called the baculovirus repeated ORFs (bro) family. Computer-assisted analysis revealed that BRO-A does not contain the conserved Bro-N domain in its N-terminus, but contains a single-stranded DNA binding motif near its C-terminus. Transcriptional analysis indicated that bro-a transcript initiated within a baculovirus consensus early start site motif (ATCAGT). In vitro the bro-a gene transcript was present at 12 h post infection (p.i.) and remained detectable up to 96 h p.i. Western blot analysis of BRO-A expression in SpltMNPV infected S. litura cells showed that it was present from 72 h to 96 h p.i. in vitro. Analysis of extracts from nuclear structures indicated that BRO-A could not be extracted with DNA or proteins which bind to DNA.

  14. Identification of a new Bombyx mori nucleopolyhedrovirus and analysis of its bro gene family.

    PubMed

    Zhou, Jing-Bo; Li, Xing-Qi; De-Eknamkul, Wanchai; Suraporn, Siripuk; Xu, Jia-Ping

    2012-06-01

    The highly pathogenic Bombyx mori nucleopolyhedrovirus (BmNPV) has caused severe damages to sericulture in many countries, and the relationship between the pathogenicity of various BmNPV strains and their geographical evolution has been the topic of our interest. In this study, we isolated a new BmNPV strain from Thailand (BmNPV-Thai), based on the sequences of its conservative genes p10, p35, polh, egt and vp39. The BmNPV-Thai appears to have baculovirus repeated ORF (bro) genes different from four other well-known BmNPV strains of China (GD, CQ1), Japan (T3), and France (SC7); It only has bro-a, bro-c, and bro-d, but not bro-b and bro-e genes. These bro genes are localized only in the two subgroups highly homologous to their counterparts and their encoded BRO proteins differ mainly at their N-terminal amino acid residues. Phylogenetic analysis indicates that the evolution of the bro genes of the five BmNPV strains is not obviously associated with their geographic locations.

  15. Interaction of Bombyx mori nucleopolyhedrovirus BRO-A and host cell protein laminin.

    PubMed

    Kang, W K; Imai, N; Suzuki, M; Iwanaga, M; Matsumoto, S; Zemskov, E A

    2003-01-01

    The Bombyx mori nucleopolyhedrovirus (BmNPV) contains five baculovirus repeated ORF ( bro) genes, all of which are expressed as delayed early genes. We have recently reported that BmNPV BRO proteins, specially BRO-A and BRO-C, contain a nucleic acid binding activity and are involved in nucleosome structures in nuclei of infected cells. To further understand the function of bro-a gene, we looked for factors interacting with BmNPV BRO-A using the yeast two-hybrid system. Fifteen clones obtained from a cDNA library of mock-infected cells and one from a library prepared at 2 h postinfection (p.i.) were found to comprise one distinct gene, which was identified as the Bombyx homolog (bLaminin) of Drosophila laminin beta1. A direct interaction between BRO-A and N-terminal region of bLaminin was demonstrated by in vitro pull-down experiments. Further pull-down assays using BmN cell extracts and anti-laminin antibodies also showed interaction of both proteins. In addition, two more clones were obtained from cDNA library of 12 h p.i. and were found to encode BRO-A itself, indicating that BRO-A forms an oligomer. Taken together, we propose that BRO-A may function as a laminin binding protein.

  16. Characterization of bro-b gene of Spodoptera litura multicapsid nucleopolyhedrovirus.

    PubMed

    Gong, Yingxue; Li, Zhaofei; Wang, Lihua; Pan, Lijing; Yang, Kai; Pang, Yi

    2003-10-01

    Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) ORF125 (designated as the bro-b gene) is a member of the unique multigene family called the baculovirus repeated ORFs (bro) family. Computer-assisted analysis revealed that BRO-B contains a conserved Bro-N domain in its N-terminus and a single-stranded DNA (ssDNA) binding motif in the middle region. In vitro the bro-b gene transcription was present at 12 h post-infection (p.i.) and remained detectable up to 96 h p.i. Western blot analysis of BRO-B expression with an antiserum made against 6xHis tagged BRO-B expressed in Escherichia coli showed that it was present from 12 h through 96 h p.i. in vitro. Structural localization revealed that BRO-B could be found in the nucleocapsid components of both occlusion-derived virus (ODV) and budded virus (BV). Immunofluorescence analysis indicated that BRO-B is located only in the nucleus of infected S. litura cells. Furthermore, Western blot analysis indicated that BRO-B was associated with nuclear structures. These results suggested that BRO-B might be a nuclear-associated protein.

  17. Diversity, distribution, and mobility of bro gene sequences in Bombyx mori nucleopolyhedrovirus.

    PubMed

    López Ferber, M; Argaud, O; Croizier, L; Croizier, G

    2001-06-01

    Genetic differences between strains of a baculovirus are often limited to some restriction sites, short DNA deletions or absence of some nonessential genes. The recently coined bro gene family, represents a new major source of intraspecific variability. A comparison between two bro gene sets of Bombyx mori nucleopolyhedroviruses (NPV) shows that bro genes are distributed in three regions for the -T3 and -SC7 virus strains. In BmNPV T3, five bro genes are distributed in three genome locations, whereas the BmNPV SC7 strain possess a single bro copy in each region. In addition, each of the BmNPV SC7 bro genes belongs to one of the three subfamilies present in BmNPV T3. Analysis of bro copy sequences and of adjacent sequences suggests an active redistribution of sequences due to intraspecific recombination. The maintenance of one allele of each subfamily suggests that they play different roles in the viral cycle, and that they are essential.

  18. Proteomics analysis of digestive juice from silkworm during Bombyx mori nucleopolyhedrovirus infection.

    PubMed

    Hu, Xiaolong; Zhu, Min; Wang, Simei; Zhu, Liyuan; Xue, Renyu; Cao, Guangli; Gong, Chengliang

    2015-08-01

    Previous studies have analyzed the midgut transcriptome and proteome after challenge with Bombyx mori nucleopolyhedrovirus (BmNPV), however little information is available on the digestive juice proteome after BmNPV challenge. This study investigated BmNPV infection-induced protein changes in the digestive juice of silkworms using shotgun proteomics and MS sequencing. From the digestive juice of normal third-day, fifth-instar silkworm larvae, 75 proteins were identified, 44 of which were unknown; from larvae 6 h after inoculation with BmNPV, 106 proteins were identified, of which 39 were unknown. After BmNPV challenge, more secreted proteins appeared that had antiviral and digestive features. GO annotation analysis clustered most proteins in the lumen into catalytic, binding, and metabolic processes. Numerous proteins were reported to have BmNPV interactions. Hsp70 protein cognate, lipase-1, and chlorophyllide A-binding protein precursor were upregulated significantly after BmNPV challenge. Levels of trypsin-like serine protease, beta-1,3-glucanase, catalase, and serine protease transcripts decreased or were not significantly change after BmNPV challenge. Taken together, these findings provided insights into the interaction between host and BmNPV and revealed potential functions of digestive juice after per os BmNPV infection.

  19. Conservation of DNA photolyase genes in group II nucleopolyhedroviruses infecting plusiine insects.

    PubMed

    Xu, Fang; Vlak, Just M; van Oers, Monique M

    2008-09-01

    DNA photolyase genes (phr) encode photoreactive enzymes, which are involved in the repair of UV-damaged DNA. Cyclobutane pyrimidine dimer (CPD) specific photolyase genes are present in nucleopolyhedroviruses isolated from Chrysodeixis chalcites (ChchNPV) and Trichoplusia ni (TnSNPV), insects belonging to the Plusiinae (Noctuidae). To better understand the occurrence and evolution of these genes in baculoviruses, we investigated their possible conservation in other group II NPVs, which infect plusiine insects. A PCR based strategy using degenerate phr-specific primers was designed to detect and analyze possible photolyase genes. Six additional Plusiinae-infecting NPVs were analyzed and all, except Thysanoplusia oricalcea NPV A28-1, which is a group I NPV, contained one or more phr-like sequences. Phylogenetic analysis revealed that all photolyase genes of the tested Plusiinae-infecting baculoviruses group in a single clade, separated into three subgroups. The phylogeny of the polyhedrin sequences of these viruses confirmed that the analyzed viruses also formed a single clade in group II NPVs. We hypothesize that all plusiine group II NPVs contain one or more photolyase genes and that these have a common ancestor.

  20. Genome Sequencing and Analysis of Catopsilia pomona nucleopolyhedrovirus: A Distinct Species in Group I Alphabaculovirus

    PubMed Central

    Wang, Jun; Zhu, Zheng; Zhang, Lei; Hou, Dianhai; Wang, Manli; Arif, Basil; Kou, Zheng; Wang, Hualin; Deng, Fei; Hu, Zhihong

    2016-01-01

    The genome sequence of Catopsilia pomona nucleopolyhedrovirus (CapoNPV) was determined by the Roche 454 sequencing system. The genome consisted of 128,058 bp and had an overall G+C content of 40%. There were 130 hypothetical open reading frames (ORFs) potentially encoding proteins of more than 50 amino acids and covering 92% of the genome. Among all the hypothetical ORFs, 37 baculovirus core genes, 23 lepidopteran baculovirus conserved genes and 10 genes conserved in Group I alphabaculoviruses were identified. In addition, the genome included regions of 8 typical baculoviral homologous repeat sequences (hrs). Phylogenic analysis showed that CapoNPV was in a distinct branch of clade “a” in Group I alphabaculoviruses. Gene parity plot analysis and overall similarity of ORFs indicated that CapoNPV is more closely related to the Group I alphabaculoviruses than to other baculoviruses. Interesting, CapoNPV lacks the genes encoding the fibroblast growth factor (fgf) and ac30, which are conserved in most lepidopteran and Group I baculoviruses, respectively. Sequence analysis of the F-like protein of CapoNPV showed that some amino acids were inserted into the fusion peptide region and the pre-transmembrane region of the protein. All these unique features imply that CapoNPV represents a member of a new baculovirus species. PMID:27166956

  1. Proteomic analyses of baculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus budded and occluded virus.

    PubMed

    Braconi, Carla Torres; Ardisson-Araújo, Daniel Mendes Pereira; Paes Leme, Adriana Franco; Oliveira, Juliana Velasco de Castro; Pauletti, Bianca Alves; Garcia-Maruniak, Alejandra; Ribeiro, Bergmann Morais; Maruniak, James E; Zanotto, Paolo Marinho de Andrade

    2014-04-01

    Baculoviruses infect insects, producing two distinct phenotypes during the viral life cycle: the budded virus (BV) and the occlusion-derived virus (ODV) for intra- and inter-host spread, respectively. Since the 1980s, several countries have been using Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) as a biological control agent against the velvet bean caterpillar, A. gemmatalis. The genome of AgMNPV isolate 2D (AgMNPV-2D) carries at least 152 potential genes, with 24 that possibly code for structural proteins. Proteomic studies have been carried out on a few baculoviruses, with six ODV and two BV proteomes completed so far. Moreover, there are limited data on virion proteins carried by AgMNPV-2D. Therefore, structural proteins of AgMNPV-2D were analysed by MALDI- quadrupole-TOF and liquid chromatography MS/MS. A total of 44 proteins were associated with the ODV and 33 with the BV of AgMNPV-2D. Although 38 structural proteins were already known, we found six new proteins in the ODV and seven new proteins carried by the AgMNPV-2D BV. Eleven cellular proteins that were found on several other enveloped viruses were also identified, which are possibly carried with the virion. These findings may provide novel insights into baculovirus biology and their host interaction. Moreover, our data may be helpful in subsequent applied studies aiming to improve AgMNPV use as a biopesticide and a biotechnology tool for gene expression or delivery.

  2. Identification and characterization of odv-e25 of Spodoptera litura multicapsid nucleopolyhedrovirus.

    PubMed

    Li, Zhaofei; Pan, Lijin; Yu, Hang; Li, Sainan; Zhang, Guoping; Pang, Yi

    2006-02-01

    Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) odv-e25 is 684 bp long, potentially encoding 227 amino acids with a predicted molecular weight of 24.9 kDa. Homology analysis indicated that SpltMNPV ODV-E25 has 35-65% amino acid identity with that of other known baculoviruses. RT-PCR results revealed that the odv-e25 is transcribed actively at the late stage of infection and the mRNA start site was mapped within a consensus baculovirus late promoter sequence (TTAAG). Western blot analysis of odv-e25 expression with an antiserum made against 6 x His tagged ODV-E25 expressed in Escherichia coli indicated that it was present as a doublet of approximately 27 kDa from 24 h through 96 h in SpltMNPV-infected Spli-221 cells. Similar results were seen on Western blots of Spodoptera exigua (Se)MNPV-infected Se301 cells. Immunofluorescence analysis showed that ODV-E25 was predominantly present in the cytoplasm of SpltMNPV-infected cells and localized to the envelopes of occlusion-derived virus.

  3. Autographa californica multiple nucleopolyhedrovirus odv-e66 is an essential gene required for oral infectivity.

    PubMed

    Xiang, Xingwei; Chen, Lin; Hu, Xiaolong; Yu, Shaofang; Yang, Rui; Wu, Xiaofeng

    2011-06-01

    Autographa californica multiple nucleopolyhedrovirus (AcMNPV) odv-e66 is a core gene and encodes an occlusion-derived virus (ODV)-specific envelope protein, ODV-E66. The N-terminal 23 amino acid of the envelope protein ODV-E66 are sufficient to direct native and fusion proteins to induced membrane microvesicles and the viral envelope during infection with AcMNPV. In this study, an odv-e66-knockout bacmid can not express N-terminal hydrophobic domains was constructed via homologous recombination in Escherichia coli. The odv-e66 deletion had no effect on budded virus (BV) production and viral DNA replication in infected Sf9 cells. Larval bioassays demonstrated that injection of odv-e66 deletion BV into the hemocoel could kill P. xylostella larvae as efficiently as repaired and control viruses; however, odv-e66 deletion mutant resulted in a 50% lethal dose that was 10(3) higher than that of the repaired and control viruses when inoculated per os. These results indicated that ODV-E66 envelope protein most likely played an important role in the oral infectivity of AcMNPV, but is not essential for virus replication.

  4. Molecular Characterization of a Nucleopolyhedrovirus Newly Isolated from Ophiusa disjungens in China.

    PubMed

    Lin, Tong; Liu, Li; Chang, Runlei; Lang, Guojun; Xu, Wen

    2012-12-01

    Ophiusa disjungens nucleopolyhedrovirus (OpdiNPV) was newly found in Guangdong Province, China. Using BamHI, EcoRI, HindIII, PstI, XhoI, XbaI digestion, the size of the OpdiNPV genome was estimated to be 92,000 base pair. The pstI-G genomic fragment of OpdiNPV was cloned and sequenced. Searches of databases identified at least four open reading frames (ORFs) within this fragment. These ORFs encode odv-e66 (EU 623602), p87/vp80 (EU 732665), odv-ec43 (EU617337) and ac108 gene (EU 732666) respectively. The phylogenetic tree of NPVs based on the combined sequences of odv-ec43 and ac108 indicated that OpdiNPV was most closely related to Mamestra configurata NPV-A and Mamestra configurata NPV-B. The characterization of OpdiNPV in this paper would provide better understanding molecular properties of this virus and be helpful in the development of the newly isolated virus as a biopesticide or an engineered pesticide to control more species of insect pests.

  5. Protein composition of the occlusion derived virus of Chrysodeixis chalcites nucleopolyhedrovirus.

    PubMed

    Xu, Fang; Ince, Ikbal Agah; Boeren, Sjef; Vlak, Just M; van Oers, Monique M

    2011-06-01

    Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV) is a group II NPV and its genome has 151 predicted open reading frames. In this study, the protein composition of ChchNPV occlusion derived virus (ODV) was determined by LC-MS/MS. Fifty-three proteins were identified in ChchNPV ODV particles. One ODV-protein is encoded by a gene so far unique to ChchNPV (Chch105). The two DNA photolyases PHR1 and PHR2, which are characteristic for ChchNPV and thought to be involved in repairing UV damage in viral DNA, were not detected in the ODVs. Comparison of the ODV proteins identified in ChchNPV and in three other baculoviruses enabled the identification of ten conserved ODV proteins (ODV-E18, ODV-E56, ODV-EC27, ODV-EC43, P6.9, P33, P49, P74, GP41, and VP39). In addition, the baculovirus per os infectivity factors PIF1, PIF2 and PIF3 were all detected in ChchNPV and these should be considered as conserved ODV proteins as well as they are absolutely required for oral infection. With the LC-MS/MS method used 22 viral proteins were detected, which were not identified as ODV proteins in previous studies.

  6. The structural protein ODV-EC27 of Autographa californica nucleopolyhedrovirus is a multifunctional viral cyclin.

    PubMed

    Belyavskyi, M; Braunagel, S C; Summers, M D

    1998-09-15

    Two major characteristics of baculovirus infection are arrest of the host cell at G2/M phase of the cell cycle with continuing viral DNA replication. We show that Autographa californica nucleopolyhedrovirus (AcMNPV) encodes for a multifunctional cyclin that may partially explain the molecular basis of these important characteristics of AcMNPV (baculovirus) infection. Amino acids 80-110 of the viral structural protein ODV-EC27 (-EC27) demonstrate 25-30% similarity with cellular cyclins within the cyclin box. Immunoprecipitation results using antibodies to -EC27 show that -EC27 can associate with either cdc2 or cdk6 resulting in active kinase complexes that can phosphorylate histone H1 and retinoblastoma protein in vitro. The cdk6-EC27 complex also associates with proliferating cell nuclear antigen (PCNA) and we demonstrate that PCNA is a structural protein of both the budded virus and the occlusion-derived virus. These results suggest that -EC27 can function as a multifunctional cyclin: when associated with cdc2, it exhibits cyclin B-like activity; when associated with cdk6, the complex possesses cyclin D-like activity and binds PCNA. The possible roles of such a multifunctional cyclin during the life cycle of baculovirus are discussed, along with potential implications relative to the expression of functionally authentic recombinant proteins by using baculovirus-infected cells.

  7. Functional analysis of Spodoptera frugiperda nucleopolyhedrovirus late expression factors in Sf9 cells.

    PubMed

    Berretta, Marcelo F; López, M Gabriela; Taboga, Oscar; Sciocco-Cap, Alicia; Romanowski, Víctor

    2013-02-01

    We used transient expression assays to assess the function of the baculovirus Spodoptera frugiperda M nucleopolyhedrovirus (SfMNPV) homologs of Autographa californica MNPV (AcMNPV) factors involved in late gene expression (lefs), in the Sf9 insect cell-line, which is permissive for both viruses. It is well-established that nineteen AcMNPV lefs support optimal levels of activity from a late promoter-reporter gene cassette in this assay. A subgroup of SfMNPV lefs predicted to function in transcription-specific events substituted the corresponding AcMNPV lefs very efficiently. When all SfMNPV lefs were assayed, including replication lefs, activity was low, but addition of two AcMNPV lefs not encoded in SfMNPV genome, resulted in augmented reporter activity. SfMNPV IE-1 was able to activate an early promoter cis-linked to an hr-derived element from SfMNPV but not from AcMNPV. However, the level of early promoter activation with SfMNPV IE-1 was lower compared to AcMNPV IE-1.

  8. Immobilization of foreign protein into polyhedra of Bombyx mori nucleopolyhedrovirus (BmNPV)*

    PubMed Central

    Xiang, Xing-wei; Yang, Rui; Chen, Lin; Hu, Xiao-long; Yu, Shao-fang; Cao, Cui-ping; Wu, Xiao-feng

    2012-01-01

    In the late phase of Bombyx mori nucleopolyhedrovirus (BmNPV) infection, a large amount of polyhedra appear in the infected cell nucleolus, these polyhedra being dense protein crystals protecting the incorporated virions from the harsh environment. To investigate whether the foreign protein could be immobilized into the polyhedra of BmNPV, two recombinant baculoviruses were generated by a novel BmNPV polyhedrin-plus (polh+) Bac-to-Bac system, designated as vBmBac(polh+)-enhanced green fluorescent protein (EGFP) and vBmBac(polh+)-LacZ, which can express the polyhedrin and foreign protein simultaneously. Light microscopy analysis showed that all viruses produced polyhedra of normal appearance. Green fluorescence can be apparently detected on the surface of the vBmBac(polh+)-EGFP polyhedra, but not the BmNPV polyhedra. Fluorescence analysis and anti-desiccation testing confirmed that EGFP was embedded in the polyhedra. As expected, the vBmBac(polh+)-LacZ polyhedra contained an amount of LacZ and had a higher β-galactosidase activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were also performed to verify if the foreign proteins were immobilized into polyhedra. This study provides a new inspiration for efficient preservation of useful proteins and development of new pesticides with toxic proteins. PMID:22302424

  9. Bm65 is essential for the propagation of Bombyx mori nucleopolyhedrovirus.

    PubMed

    Tang, Qi; Li, Guohui; Yao, Qin; Chen, Liang; Feng, Fan; Yuan, Yi; Chen, Keping

    2013-01-01

    Orf65 (Bm65) of Bombyx mori nucleopolyhedrovirus (BmNPV) is a highly conserved gene that encodes an unknown 104-amino acid protein. In the present study, we have shown the role of Bm65 in the baculovirus life cycle. 5'-RACE analysis showed that the transcription start site of Bm65 was 14 nucleotides upstream of the start codon ATG. The transcription profile of Bm65 was detected from 6 to 72 h postinfection (p. i.) by RT-PCR. A Bm65-knockout bacmid was constructed by homologous recombination to characterize the role of Bm65 in viral life cycle. Fluorescence microscopy showed that Bm65-knockout virus was unable to generate infectious budded virus in BmN cells. Furthermore, quantitative real-time PCR analysis demonstrated that Bm65 deletion did not affect the viral DNA replication. To conclude, Bm65 is essential for the propagation of BmNPV, but is unnecessary for the replication of viral DNA.

  10. Evidence for Nucleic Acid Binding Ability and Nucleosome Association of Bombyx mori Nucleopolyhedrovirus BRO Proteins

    PubMed Central

    Zemskov, Evgueni A.; Kang, WonKyung; Maeda, Susumu

    2000-01-01

    The Bombyx mori nucleopolyhedrovirus (BmNPV) genome contains five related members of the bro gene family, all of which are actively expressed in infected BmN cells. Although their functions are unknown, their amino acid sequences contain a motif found in all known viral and prokaryotic single-stranded DNA binding proteins. To determine if they bind to nucleic acids, we fractionated the nuclei of BmNPV-infected BmN cells using a histone extraction protocol. We detected BRO-A, BRO-C, and BRO-D in the histone H1 fraction using anti-BRO antibodies. Micrococcal nuclease treatment released these BRO proteins from the chromatin fraction, suggesting their involvement in nucleosome structures. Chromatographic fractionation showed that BRO-A and/or BRO-C interacted with core histones. Expression of partial sequences of BRO-A proved that the N-terminal 80 amino acid residues were required for DNA binding activity. We also demonstrated that BmNPV BRO proteins underwent phosphorylation and ubiquitination followed by proteasome degradation, which may explain their distribution in the cytoplasm as well as the nucleus. We propose that BRO-A and BRO-C may function as DNA binding proteins that influence host DNA replication and/or transcription. PMID:10888617

  11. Does Florivory by Helicoverpa zea (Lepidoptera: Noctuidae) Cause Yield Loss in Soybeans?

    PubMed

    Reisig, Dominic; Suits, Rachel; Burrack, Hannah; Bacheler, Jack; Dunphy, Jim E

    2017-04-01

    Helicoverpa zea (Boddie), corn earworm, is a damaging insect pest of many crops, including soybeans. An economic threshold for soybeans during the pod-filling stages exists to prevent economic damage to seeds. However, the impact of florivory (flower feeding) by H. zea larvae on seed yield is poorly understood and there is no economic threshold for flowering-stage soybeans. Four small plot experiments were conducted in North Carolina during 2011 and 2012 to assess the impact of H. zea feeding during the flowering stages of determinate soybeans on various yield components. Helicoverpa zea densities were manipulated with insecticides and various planting dates of soybeans and monitored weekly. Helicoverpa zea naturally infested the plots after flowering began and were allowed to feed until R3; they were eliminated from all plots from R3 to maturity. In some sites, H. zea densities exceeded the podding economic threshold during the flowering stages, but yield did not differ among treatments. During 2012, florivory from H. zea was measured directly by counting injured flowers. There was a negative yield relationship between both injured flower number and cumulative flower number. Moreover, H. zea densities were related to both a decrease in cumulative flowers and an increase in injured flowers, even though a direct linkage between H. zea density and yield loss was not observed. Without knowing the preferred tissue types and performance of early-instar larvae on soybeans, it is possible that H. zea density may not be the best measurement for developing an economic threshold in flowering soybeans. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  12. The Trichoplusia ni single nucleopolyhedrovirus tn79 gene encodes a functional sulfhydryl oxidase enzyme that is able to support the replication of Autographa californica multiple nucleopolyhedrovirus lacking the sulfhydryl oxidase ac92 gene

    PubMed Central

    Clem, Stian A.; Wu, Wenbi; Lorena Passarelli, A.

    2014-01-01

    The Autographa californica multiple nucleopolyhedrovirus ac92 is a conserved baculovirus gene with homology to flavin adenine dinucleotide-linked sulfhydryl oxidases. Its product, Ac92, is a functional sulfhydryl oxidase. Deletion of ac92 results in almost negligible levels of budded virus (BV) production, defects in occlusion-derived virus (ODV) co-envelopment and their inefficient incorporation into occlusion bodies. To determine the role of sulfhydryl oxidation in the production of BV, envelopment of nucleocapsids, and nucleocapsid incorporation into occlusion bodies, the Trichoplusia ni single nucleopolyhedrovirus ortholog, Tn79, was substituted for ac92. Tn79 was found to be an active sulfhydryl oxidase that substituted for Ac92, resulting in the production of infectious BV, albeit about 10-fold less than an ac92-containing virus. Tn79 rescued defects in ODV morphogenesis caused by a lack of ac92. Active Tn79 sulfhydryl oxidase activity is required for efficient BV production, ODV envelopment, and their subsequent incorporation into occlusion bodies in the absence of ac92. PMID:25010286

  13. Characterization of the interaction between P143 and LEF-3 from two different baculovirus species: Choristoneura fumiferana nucleopolyhedrovirus LEF-3 can complement Autographa californica nucleopolyhedrovirus LEF-3 in supporting DNA replication.

    PubMed

    Chen, Tricia; Sahri, Daniela; Carstens, Eric B

    2004-01-01

    The baculovirus protein P143 is essential for viral DNA replication in vivo, likely as a DNA helicase. We have demonstrated that another viral protein, LEF-3, first described as a single-stranded DNA binding protein, is required for transporting P143 into the nuclei of insect cells. Both of these proteins, along with several other early viral proteins, are also essential for DNA replication in transient assays. We now describe the identification, nucleotide sequences, and transcription patterns of the Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) homologues of p143 and lef-3 and demonstrate that CfMNPV LEF-3 is also responsible for P143 localization to the nucleus. We predicted that the interaction between P143 and LEF-3 might be critical for cross-species complementation of DNA replication. Support for this hypothesis was generated by substitution of heterologous P143 and LEF-3 between two different baculovirus species, Autographa californica nucleopolyhedrovirus and CfMNPV, in transient DNA replication assays. The results suggest that the P143-LEF-3 complex is an important baculovirus replication factor.

  14. Impact of deletion of the Lymantria dispar nucleopolyhedrovirus PEP gene on viral potency: expression of the green fluorescent protein prevents larval liquefaction

    Treesearch

    David S. Bischoff; James M. Slavicek

    1999-01-01

    The Lymantria dispar multicapsid nucleopolyhedrovirus (LdMNPV) is an effective biological control agent of the gypsy moth, L. dispar, but is not in general use because the high cost of production limits availability. In an effort to generate a more cost efficient LdMNPV biopesticide, two...

  15. Deletion of the Lymantria dispar multicapsid nucleopolyhedrovirus ecdysteroid UDP-glucosyl transferase gene enhances viral killing speed in the last instar of the gypsy moth

    Treesearch

    James M. Slavicek; Holly J.R. Popham; C.I. Riegel

    1999-01-01

    The Lymantria dispar multicapsid nucleopolyhedrovirus (LdMNPV) is used on a limited basis as a gypsy moth (L. dispar) control agent. In an effort to improve the efficacy (i.e., killing speed) of the LdMNPV, we generated a recombinant viral strain (vEGT-) that does not produce the enzyme ecdysteroid UDP-glucosyltransferase (EGT). We...

  16. Phenotypic and genetic analysis of Lymantria dispar nucleopolyhedrovirus few polyhedra mutants: Mutations in the 25K FP gene may be caused by DNA replication errors

    Treesearch

    David S. Bischoff; James M. Slavicek

    1997-01-01

    We previously demonstrated that polyhedron formation (PF) mutants arise at a high frequency during serial passage of the Lymantria dispar nucleopolyhedrovirus (LdMNPV) in the L. dispar 652Y cell line (J.M. Slavicek, N. Hayes-Plazolles, and M.E. Kelly, Biol. Control 5:251-261, 1995). Most of these PF mutants...

  17. Identification of a Lymantria dispar nucleopolyhedrovirus isolate that does not accumulate few-polyhedra mutants during extended serial passage in cell culture

    Treesearch

    James M. Slavicek; Nancy Hayes-Plazolles; Mary Ellen. Kelly

    2001-01-01

    During Lymantria dispar multinucleocapsid nucleopolyhedrovirus (LdMNPV) replication in cell culture, few-polyhedra (FP) mutants arise at a high frequency and become predominant after only a few serial passages. The formation of LdMNPV FP mutants was an impediment to successful production of polyhedra in cell culture bioreactors. We have isolated and...

  18. Slow cell infection, inefficient primary infection and inability to replicate in fat body determine host-range of Thysanoplusia orichalcea M Nucleopolyhedrovirus (ThorMNPV)

    USDA-ARS?s Scientific Manuscript database

    An enhanced green fluorescence protein (EGFP) gene expression cassette was inserted at the gp37 locus of Thysanoplusia orichacea M nucleopolyhedrovirus (ThorMNPV) to produce vThGFP to study host-range mechanisms. Using EGFP to monitor infection in vitro, many cell lines showed EGFP expression sugges...

  19. Screening of candidate proteins interacting with IE-2 of Bombyx mori nucleopolyhedrovirus.

    PubMed

    Wu, Yejun; Wu, Yu; Wu, Yan; Tang, Hui; Wu, Huiling; Zhang, Guozheng; Wang, Wenbing

    2013-10-01

    IE-2 of Bombyx mori nucleopolyhedrovirus (BmNPV) has been shown to play important roles in baculovirus infection, which are involved in gene expression and viral replication. However, the mechanism remains unknown. In this paper, by TargetP software, four genes, i.e.-2, odv-e26, odv-e56 and BmNPV-gp101 (Ac-orf116) of BmNPV and Autographa californica multiple NPV (AcMNPV) were predicted to be located in mitochondria. By BLAST tool using BmNPV IE-2 protein sequence, 14 NPVs were found to have IE-2 homologues in GenBank, and most of them were predicted to be located in mitochondria, except for that of Antheraea pernyi NPV (AnpeNPV) and Anticarsia gemmatalis NPV (AngeNPV). To observe the subcellular localization of BmNPV IE-2, a recombinant virus overexpressed the IE-2 and eGFP fusion protein was constructed. In infected BmN cells, the fluorescence specifically enriched in the cellular mitochondria. This evidence was accordant with the prediction. Further, Pull-down assay was used to select protein candidates interacting with IE-2 in B. mori cells infected with BmNPV. Of several isolated protein components, sixteen candidates were identified by MALDI-TOF mass-spectrometry, eight baculoviral proteins (ALK-EXO, F protein, IAP-1, LEF-3, LEF-9, ODV-NC42, TLP, and VP39), and eight proteins from B. mori (Actin, ADP/ATP translocase, ATP synthase subunit beta, Beta-tubulin, DNA topoisomerase 2, Histone H4, Soluble guanylyl cyclae alpha-1 subunit, Transketolase). From the functional point of view, most of these proteins were generally divided into two groups, mitochondrial interaction proteins and viral DNA replication proteins. These results implied that the IE-2 had multiple functions involved in regulating viral gene expression, viral replication and also as a component of mitochondrial factors to regulate the cellular energy supply and apoptosis.

  20. A Hypothetical Model of Crossing Bombyx mori Nucleopolyhedrovirus through Its Host Midgut Physical Barrier

    PubMed Central

    Cheng, Yang; Wang, Xue-Yang; Hu, Hao; Killiny, Nabil; Xu, Jia-Ping

    2014-01-01

    Bombyx mori nucleopolyhedrovirus (BmNPV) is a primary pathogen of silkworm (B. mori) that causes severe economic losses each year. However, the molecular mechanisms of silkworm-BmNPV interactions, especially the silkworm proteins that can interact with the virus, are still largely unknown. In this study, the total and membrane proteins of silkworm midguts were displayed using one- and two-dimensional electrophoresis. A virus overlay assay was used to detect B. mori proteins that specifically bind to BmNPV particles. Twelve proteins were located and identified using mass spectrometry, and the different expression of the corresponding genes in BmNPV susceptible and resistant silkworm strains also indicated their involvement in BmNPV infection. The 12 proteins are grouped based on their potential roles in viral infection, for example, endocytosis, intracellular transportation, and host responses. Based on these results, we hypothesize the following: I) vacuolar ATP synthase catalytic subunit A and subunit B may be implicated in the process of the membrane fusion of virus and the release of the nucleocapsid into cytoplasm; II) actin, enolase and phosphoglycerate kinase are cytoskeleton associated proteins and may play an important role in BmNPV intracellular transportation; III) mitochondrial prohibitin complex protein 2, ganglioside-induced differentiation-associated protein, calreticulin, regucalcin-like isoform X1 and 60 kDa heat shock protein are involved in cell apoptosis regulation during BmNPV infection in larvae midguts; IV) ribosomal P0 may be associated with BmNPV infection by regulating gene expression of BmNPV; V) arginine kinase has a role in the antiviral activities against BmNPV. Our work should prove informative by providing multiple protein targets and a novel direction to investigate the molecular mechanisms of the interactions between silkworms and BmNPV. PMID:25502928

  1. Isolation and characterization of plaque-purified strains of Malacosoma disstria Nucleopolyhedrovirus.

    PubMed

    Erlandson, Martin A; Baldwin, Doug; Haveroen, Melissa; Keddie, B Andrew

    2006-03-01

    Seven plaque-purified genotypic variants or strains, derived from a previously described field isolate of the Malacosoma disstria Nucleopolyhedrovirus (MadiNPV) from Alberta populations of forest tent caterpillar, were characterized based on distinctive restriction endonuclease fragment patterns. Two strains, MadiNPV-pp3 and MadiNPV-pp11, were selected for further characterization, as they represented strains producing high and low budded virus (BV) titres, respectively, in the M. disstria cell line UA-Md203. Analysis of restriction endonuclease fragment profiles indicated the genomes differed significantly in size, 133.8 +/- 2.4 kb for MadiNPV-pp3 and 118.1 +/- 3.5 kb for MadiNPV-pp11. These strains were characterized based on their BV production in three different cell lines derived from M. disstria haemocytes. Compared with MadiNPV-pp11, MadiNPV-pp3 produced two- to three-fold more BVs in UA-Md203 and 210 other cell lines; however, BV production was only marginally higher for MadiNPV-pp3 in the UA-Md221 cell line. Similarly, the yield of polyhedral inclusion bodies was significantly higher for MadiNPV-pp3 in UA-Md203 and 210 cell lines than for MadiNPV-pp11 but not in the UA-Md221 cell line. This data, although derived from a limited number of cell lines, suggested MadiNPV-pp3 may have a broader tissue tropism than MadiNPV-pp11.

  2. Selection and Characterization of Autographa californica Multiple Nucleopolyhedrovirus DNA Polymerase Mutations

    PubMed Central

    Feng, Guozhong; Thumbi, David K.; de Jong, Jondavid; Hodgson, Jeffrey J.; Arif, Basil M.; Doucet, Daniel

    2012-01-01

    Autographa californica multiple nucleopolyhedrovirus (AcMNPV) DNA polymerase (DNApol) is essential for viral DNA replication. AcMNPV mutants resistant to aphidicolin, a selective inhibitor of viral DNA replication, and abacavir, an efficacious nucleoside analogue with inhibitory activity against reverse transcriptase, were selected by the serial passage of the parental AcMNPV in the presence of increasing concentrations of aphidicolin or abacavir. These drug-resistant mutants had either a single (C543R) (aphidicolin) or a double (C543R and S611T) (abacavir) point mutation within conserved regions II and III. To confirm the role of these point mutations in AcMNPV DNA polymerase, a dnapol knockout virus was first generated, and several repair viruses were constructed by transposing the dnapol wild-type gene or ones containing a single or double point mutation into the polyhedrin locus of the dnapol knockout bacmid. The single C543R or double C543R/S611T mutation showed increased resistance to both aphidicolin and abacavir and, even in the absence of drug, decreased levels of virus and viral DNA replication compared to the wild-type repair virus. Surprisingly, the dnapol mutant repair viruses led to the generation of occlusion-derived viruses with mostly single and only a few multiple nucleocapsids in the ring zone and within polyhedra. Thus, these point mutations in AcMNPV DNA polymerase increased drug resistance, slightly compromised virus and viral DNA replication, and influenced the viral morphogenesis of occlusion-derived virus. PMID:23035236

  3. Functional Importance of Deletion Mutant Genotypes in an Insect Nucleopolyhedrovirus Population

    PubMed Central

    Simón, Oihane; Williams, Trevor; López-Ferber, Miguel; Caballero, Primitivo

    2005-01-01

    A Nicaraguan isolate of a nucleopolyhedrovirus (SfNIC) that attacks the fall armyworm, Spodoptera frugiperda, survives as a mixture of nine genotypes (SfNIC A to I) that all present genomic deletions, except variant B (complete genotype). Sequencing of cloned restriction fragments revealed that genotypic variants lack between 5 and 16 of the open reading frames present in a contiguous sequence of 18 kb of the SfNIC genome. The absence of oral infectivity of SfNIC-C and -D variants is related to the deletion of the pif and/or pif-2 gene, while that of SfNIC-G remains unexplained. The presence of open reading frame 10, homolog of Se030, also appeared to influence pathogenicity in certain variants. Previous studies demonstrated a significant positive interaction between genotypes B and C. We compared the median lethal concentration of single genotypes (A, B, C, D, and F) and co-occluded genotype mixtures (B+A, B+D, B+F, A+C, and F+C in a 3:1 ratio). Mixtures B+A and B+D showed increased pathogenicity, although only B+D restored the activity of the mixture to that of the natural population. Mixtures of two deletion variants (A+C and F+C) did not show interactions in pathogenicity. We conclude that minority genotypes have an important influence on the overall pathogenicity of the population. These results clearly demonstrate the value of retaining genotypic diversity in virus-based bioinsecticides. PMID:16085811

  4. In vivo recombination between two strains of the genus Nucleopolyhedrovirus in its natural host, Spodoptera exigua.

    PubMed Central

    Muñoz, D; Vlak, J M; Caballero, P

    1997-01-01

    The DNA restriction map for the enzymes BamHI, BglII, PstI, and XbaI of SeMNPV-US (Se-US), the best-studied Nucleopolyhedrovirus strain isolated from Spodoptera exigua in California, was used as a reference to construct that of SeMNPV-SP2 (Se-SP2), a closely related Spanish strain of the same virus. After coinfection of S. exigua with both the Se-US and Se-SP2 strains per os, a recombinant virus (SeMNPV-SUR1 [Se-SUR1]) was detected after one passage which quickly replaced the parental viruses. A physical map of Se-SUR1 DNA was constructed for BamHI, BglII, PstI, and XbaI and compared to that of the parental viruses, Se-US and Se-SP2. Se-SUR1 is the result of at least four crossover events between Se-US and Se-SP2 and not of selection of a minor variant in one of the parental viruses. Bioassays of the recombinant and parental strains against L2 beet armyworm larvae demonstrated that their 50% lethal dose values were not significantly different. The differences in median lethal time values are too small to explain the replacement of the parental viruses by the recombinant virus upon successive passage in vivo, although it cannot be ruled out as an explanation for the selective advantage of the recombinant strain, Se-SUR1. The consequences of the release of nonindigenous or recombinant baculovirus strains in agro-ecosystems are discussed. PMID:9251191

  5. The Pangenome of the Anticarsia gemmatalis Multiple Nucleopolyhedrovirus (AgMNPV)

    PubMed Central

    de Brito, Anderson Fernandes; Braconi, Carla Torres; Weidmann, Manfred; Dilcher, Meik; Alves, João Marcelo Pereira; Gruber, Arthur; Zanotto, Paolo Marinho de Andrade

    2016-01-01

    The alphabaculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) is the world’s most successful viral bioinsecticide. Through the 1980s and 1990s, this virus was extensively used for biological control of populations of Anticarsia gemmatalis (Velvetbean caterpillar) in soybean crops. During this period, genetic studies identified several variable loci in the AgMNPV; however, most of them were not characterized at the sequence level. In this study we report a full genome comparison among 17 wild-type isolates of AgMNPV. We found the pangenome of this virus to contain at least 167 hypothetical genes, 151 of which are shared by all genomes. The gene bro-a that might be involved in host specificity and carrying transporter is absent in some genomes, and new hypothetical genes were observed. Among these genes there is a unique rnf12-like gene, probably implicated in ubiquitination. Events of gene fission and fusion are common, as four genes have been observed as single or split open reading frames. Gains and losses of genomic fragments (from 20 to 900 bp) are observed within tandem repeats, such as in eight direct repeats and four homologous regions. Most AgMNPV genes present low nucleotide diversity, and variable genes are mainly located in a locus known to evolve through homologous recombination. The evolution of AgMNPV is mainly driven by small indels, substitutions, gain and loss of nucleotide stretches or entire coding sequences. These variations may cause relevant phenotypic alterations, which probably affect the infectivity of AgMNPV. This work provides novel information on genomic evolution of the AgMNPV in particular and of baculoviruses in general. PMID:26615220

  6. The Pangenome of the Anticarsia gemmatalis Multiple Nucleopolyhedrovirus (AgMNPV).

    PubMed

    Brito, Anderson Fernandes de; Braconi, Carla Torres; Weidmann, Manfred; Dilcher, Meik; Alves, João Marcelo Pereira; Gruber, Arthur; Zanotto, Paolo Marinho de Andrade

    2015-11-27

    The alphabaculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) is the world's most successful viral bioinsecticide. Through the 1980s and 1990s, this virus was extensively used for biological control of populations of Anticarsia gemmatalis (Velvetbean caterpillar) in soybean crops. During this period, genetic studies identified several variable loci in the AgMNPV; however, most of them were not characterized at the sequence level. In this study we report a full genome comparison among 17 wild-type isolates of AgMNPV. We found the pangenome of this virus to contain at least 167 hypothetical genes, 151 of which are shared by all genomes. The gene bro-a that might be involved in host specificity and carrying transporter is absent in some genomes, and new hypothetical genes were observed. Among these genes there is a unique rnf12-like gene, probably implicated in ubiquitination. Events of gene fission and fusion are common, as four genes have been observed as single or split open reading frames. Gains and losses of genomic fragments (from 20 to 900 bp) are observed within tandem repeats, such as in eight direct repeats and four homologous regions. Most AgMNPV genes present low nucleotide diversity, and variable genes are mainly located in a locus known to evolve through homologous recombination. The evolution of AgMNPV is mainly driven by small indels, substitutions, gain and loss of nucleotide stretches or entire coding sequences. These variations may cause relevant phenotypic alterations, which probably affect the infectivity of AgMNPV. This work provides novel information on genomic evolution of the AgMNPV in particular and of baculoviruses in general.

  7. Mechanism of enhanced Bombyx mori nucleopolyhedrovirus-resistance by titanium dioxide nanoparticles in silkworm.

    PubMed

    Xu, Kaizun; Li, Fanchi; Ma, Lie; Wang, Binbin; Zhang, Hua; Ni, Min; Hong, Fashui; Shen, Weide; Li, Bing

    2015-01-01

    The infection of Bombyx mori nucleopolyhedrovirus (BmNPV) in silkworms is often lethal. It is difficult to prevent, and its lethality is correlated with both viral particle characteristics and silkworm strains. Low doses of titanium dioxide nanoparticles (TiO2 NPs) can promote silkworm growth and improve its resistance to organophosphate pesticides. In this study, TiO2 NPs' effect on BmNPV resistance was investigated by analyzing the characteristics of BmNPV proliferation and transcriptional differences in silkworm midgut and the transcriptional changes of immunity related genes after feeding with TiO2 NPs. We found that low doses of TiO2 NPs improved the resistance of silkworm against BmNPV by 14.88-fold, with the mortalities of the experimental group and control group being 0.56% and 8.33% at 144 h, respectively. The proliferation of BmNPV in the midgut was significantly increased 72 h after infection in both experimental and control groups; the control group reached the peak at 120 h, while the experimental group took 24 more hours to reach the maximal value that was 12.63 times lower than the control, indicating that TiO2 NPs can inhibit BmNPV proliferation in the midgut. Consistently, the expression of the BmNPV-resistant gene Bmlipase-1 had the same increase pattern as the proliferation changes. Immune signaling pathway analysis revealed that TiO2 NPs inhibited the proliferation of silkworm BmNPV to reduce the activation levels of janus kinase/signal transducer and activator of transcription (JAK/STAT) and phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway, while promoting the expression of Bmakt to improve the immunity. Overall, our results demonstrate that TiO2 NPs increase silkworm resistance against BmNPV by inhibiting virus proliferation and improving immunity in silkworms.

  8. Mechanism of Enhanced Bombyx mori Nucleopolyhedrovirus-Resistance by Titanium Dioxide Nanoparticles in Silkworm

    PubMed Central

    Zhang, Hua; Ni, Min; Hong, Fashui; Shen, Weide; Li, Bing

    2015-01-01

    The infection of Bombyx mori nucleopolyhedrovirus (BmNPV) in silkworms is often lethal. It is difficult to prevent, and its lethality is correlated with both viral particle characteristics and silkworm strains. Low doses of titanium dioxide nanoparticles (TiO2 NPs) can promote silkworm growth and improve its resistance to organophosphate pesticides. In this study, TiO2 NPs’ effect on BmNPV resistance was investigated by analyzing the characteristics of BmNPV proliferation and transcriptional differences in silkworm midgut and the transcriptional changes of immunity related genes after feeding with TiO2 NPs. We found that low doses of TiO2 NPs improved the resistance of silkworm against BmNPV by 14.88-fold, with the mortalities of the experimental group and control group being 0.56% and 8.33% at 144 h, respectively. The proliferation of BmNPV in the midgut was significantly increased 72 h after infection in both experimental and control groups; the control group reached the peak at 120 h, while the experimental group took 24 more hours to reach the maximal value that was 12.63 times lower than the control, indicating that TiO2 NPs can inhibit BmNPV proliferation in the midgut. Consistently, the expression of the BmNPV-resistant gene Bmlipase-1 had the same increase pattern as the proliferation changes. Immune signaling pathway analysis revealed that TiO2 NPs inhibited the proliferation of silkworm BmNPV to reduce the activation levels of janus kinase/signal transducer and activator of transcription (JAK/STAT) and phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway, while promoting the expression of Bmakt to improve the immunity. Overall, our results demonstrate that TiO2 NPs increase silkworm resistance against BmNPV by inhibiting virus proliferation and improving immunity in silkworms. PMID:25692869

  9. Proteomic analysis of BmN cell lipid rafts reveals roles in Bombyx mori nucleopolyhedrovirus infection.

    PubMed

    Hu, Xiaolong; Zhu, Min; Liang, Zi; Kumar, Dhiraj; Chen, Fei; Zhu, Liyuan; Kuang, Sulan; Xue, Renyu; Cao, Guangli; Gong, Chengliang

    2017-04-01

    The mechanism of how Bombyx mori nucleopolyhedrovirus (BmNPV) enters cells is unknown. The primary components of membrane lipid rafts are proteins and cholesterol, and membrane lipid rafts are thought to be an active region for host-viral interactions. However, whether they contribute to the entry of BmNPV into silkworm cells remains unclear. In this study, we explored the membrane protein components of lipid rafts from BmN cells with mass spectrometry (MS). Proteins and cholesterol were investigated after establishing infection with BmNPV in BmN cells. In total, 222 proteins were identified in the lipid rafts, and Gene Ontology (GO) annotation analysis showed that more than 10% of these proteins had binding and catalytic functions. We then identified proteins that potentially interact between lipid rafts and BmNPV virions using the Virus Overlay Protein Blot Assay (VOPBA). A total of 65 proteins were analyzed with MS, and 7 were predicted to be binding proteins involved in BmNPV cellular invasion, including actin, kinesin light chain-like isoform X2, annexin B13, heat-shock protein 90, barrier-to-autointegration factor B-like and serine/arginine-rich splicing factor 1 A-like. When the cholesterol of the lipid rafts from the membrane was depleted by methyl-β-cyclodextrin (MβCD), BmNPV entry into BmN cells was blocked. However, supplying cholesterol into the medium rescued the BmNPV infection ability. These results show that membrane lipid rafts may be the active regions for the entry of BmNPV into cells, and the components of membrane lipid rafts may be candidate targets for improving the resistance of the silkworm to BmNPV.

  10. Transport of Wild-Type and Recombinant Nucleopolyhedroviruses by Scavenging and Predatory Arthropods.

    PubMed

    Lee; Fuxa

    2000-05-01

    Wild-type and recombinant nucleopolyhedroviruses (NPVs) were compared in their capability to be transported over limited distances by the predator Podisus maculiventris (Say) and scavengers Sarcophaga bullata (Parker) and Acheta domesticus (Linnaeus) in Trichoplusia ni (Hübner) larvae infesting collards in a greenhouse microcosm. Viruses tested were variants of Autographa californica (Speyer) NPV (AcNPV): wild-type virus (AcNPV.WT), AcNPV expressing a scorpion toxin (AcNPV.AaIT), and AcNPV expressing juvenile hormone esterase (AcJHE.SG). Podisus maculiventris transported AcNPV.WT and S. bullata transported AcNPV.WT and AcNPV.AaIT. Prevalence and transport of AcNPV.WT were greater than those of AcNPV.AaIT and AcJHE.SG, regardless of whether the nontarget organism carriers were present or absent. Podisus maculiventris and S. bullata transported recombinant and wild-type NPVs at a rate of up to 62.5 cm/day, and A. domesticus transported wild-type NPV at 125 cm/day. The infected host insects, T. ni, undoubtedly contributed to viral transport in the current research. In every experiment, both the wild-type and recombinant virus spread to some degree in the plots without predators or scavengers. The relative amounts of NPVs that accumulated in soil, as indicated by bioassay mortality percentages, generally exhibited spatial patterns similar to those of T. ni mortality due to NPV on the collards plants. Thus, the predator and scavengers in the current research demonstrated some capacity to transport wild-type as well as recombinant viruses at significant rates in a greenhouse microcosm.

  11. Isolation and characterization of the DNA-binding protein (DBP) of the Autographa californica multiple nucleopolyhedrovirus

    SciTech Connect

    Mikhailov, Victor S. Vanarsdall, Adam L.; Rohrmann, George F.

    2008-01-20

    DNA-binding protein (DBP) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) was expressed as an N-terminal His{sub 6}-tag fusion using a recombinant baculovirus and purified to near homogeneity. Purified DBP formed oligomers that were crosslinked by redox reagents resulting in predominantly protein dimers and tetramers. In gel retardation assays, DBP showed a high affinity for single-stranded oligonucleotides and was able to compete with another baculovirus SSB protein, LEF-3, for binding sites. DBP binding protected ssDNA against hydrolysis by a baculovirus alkaline nuclease AN/LEF-3 complex. Partial proteolysis by trypsin revealed a domain structure of DBP that is required for interaction with DNA and that can be disrupted by thermal treatment. Binding to ssDNA, but not to dsDNA, changed the pattern of proteolytic fragments of DBP indicating adjustments in protein structure upon interaction with ssDNA. DBP was capable of unwinding short DNA duplexes and also promoted the renaturation of long complementary strands of ssDNA into duplexes. The unwinding and renaturation activities of DBP, as well as the DNA binding activity, were sensitive to sulfhydryl reagents and were inhibited by oxidation of thiol groups with diamide or by alkylation with N-ethylmaleimide. A high affinity of DBP for ssDNA and its unwinding and renaturation activities confirmed identification of DBP as a member of the SSB/recombinase family. These activities and a tight association with subnuclear structures suggests that DBP is a component of the virogenic stroma that is involved in the processing of replicative intermediates.

  12. Expression and mutational analysis of Autographa californica nucleopolyhedrovirus HCF-1: functional requirements for cysteine residues.

    PubMed

    Wilson, Joyce A; Forney, Scott D; Ricci, Alessondra M; Allen, Emily G; Hefferon, Kathleen L; Miller, Lois K

    2005-11-01

    The host cell-specific factor 1 gene (hcf-1) of the baculovirus Autographa californica multiple nucleopolyhedrovirus is required for efficient virus growth in TN368 cells but is dispensable for virus replication in SF21 cells. However, the mechanism of action of hcf-1 is unknown. To begin to understand its function in virus replication we have investigated the expression and localization pattern of HCF-1 in infected cells. Analysis of virus-infected TN368 cells showed that hcf-1 is expressed at an early time in the virus life cycle, between 2 and 12 h postinfection, and localized the protein to punctate nuclear foci. Through coprecipitation experiments we have confirmed that HCF-1 self-associates into dimers or higher-order structures. We also found that overexpression of HCF-1 repressed expression from the hcf-1 promoter in transient reporter assays. Mutagenesis of cysteine residues within a putative RING finger domain in the amino acid sequence of HCF-1 abolished self-association activity and suggests that the RING domain may be involved in this protein-protein interaction. A different but overlapping set of cysteine residues were required for efficient gene repression activity. Functional analysis of HCF-1 mutants showed that the cysteine amino acids required for both self-association and gene repression activities of HCF-1 were also required for efficient late-gene expression and occlusion body formation in TN368 cells. Mutational analysis also identified essential charged and hydrophobic amino acids located between two of the essential cysteine residues. We propose that HCF-1 is a RING finger-containing protein whose activity requires HCF-1 self-association and gene repression activity.

  13. Genetic Structure of a Spodoptera frugiperda Nucleopolyhedrovirus Population: High Prevalence of Deletion Genotypes

    PubMed Central

    Simón, Oihane; Williams, Trevor; López-Ferber, Miguel; Caballero, Primitivo

    2004-01-01

    A Nicaraguan field isolate (SfNIC) of Spodoptera frugiperda nucleopolyhedrovirus was purified by plaque assay on Sf9 cells. Nine distinct genotypes, A to I, were identified by their restriction endonuclease profiles. Variant SfNIC-B was selected as the standard because its restriction profile corresponded to that of the wild-type isolate. Physical maps were generated for each of the variants. The differences between variants and the SfNIC-B standard were confined to the region between map units 9 and 32.5. This region included PstI-G, PstI-F, PstI-L, PstI-K and EcoRI-L fragments. Eight genotypes presented a deletion in their genome compared with SfNIC-B. Occlusion body-derived virions of SfNIC-C, -D and -G accounted for 41% of plaque-purified clones. These variants were not infectious per os but retained infectivity by injection into S. frugiperda larvae. Median 50% lethal concentration values for the other cloned genotypes were significantly higher than that of the wild type. The variants also differed in their speed of kill. Noninfectious variants SfNIC-C and -D lacked the pif and pif-2 genes. Infectivity was restored to these variants by plasmid rescue with a plasmid comprising both pif and pif-2. Transcription of an SfNIC-G gene was detected by reverse transcription-PCR in insects, but no fatal disease developed. Transcription was not detected in SfNIC-C or -D-inoculated larvae. We conclude that the SfNIC population presents high levels of genetic diversity, localized to a 17-kb region containing pif and pif-2, and that interactions among complete and deleted genotypic variants will likely influence the capacity of this virus to control insect pests. PMID:15345446

  14. Nucleopolyhedrovirus detection and distribution in terrestrial, freshwater, and marine habitats of Appledore Island, Gulf of Maine.

    PubMed

    Hewson, Ian; Brown, Julia M; Gitlin, Shari A; Doud, Devin F

    2011-07-01

    Viruses in aquatic ecosystems comprise those produced by both autochthonous and allochthonous host taxa. However, there is little information on the diversity and abundance of viruses of allochthonous origin, particularly from non-anthropogenic sources, in freshwater and marine ecosystems. We investigated the presence of nucleopolyhedroviruses (NPV) (Baculovirus), which commonly infect terrestrial lepidopteran taxa, across the landscape of Appledore Island, Gulf of Maine. PCR and qPCR primers were developed around a 294-bp fragment of the polyhedrin (polH) gene, which is the major constituent protein of NPV multivirion polyhedral occlusion bodies. polH was successfully amplified from several aquatic habitats, and recovered polH sequences were most similar to known lepidopteran NPV. Using quantitative PCR designed around a cluster of detected sequences, we detected polH in Appledore Island soils, supratidal freshwater ponds, nearshore sediments, near- and offshore plankton, and in floatsam. This diverse set of locations suggests that NPVs are widely dispersed along the terrestrial--marine continuum and that free polyhedra may be washed into ponds and eventually to sea. The putative hosts of detected NPVs were webworms (Hyphantria sp.) which form dense nests in late summer on the dominant Appledore Island vegetation (Prunus virginiana). Our data indicate that viruses of terrestrial origin (i.e., allochthonous viruses) may be dispersed widely in coastal marine habitats. The dispersal of NPV polH and detection within offshore net plankton (>64 μm) demonstrates that terrestrial viruses may interact with larger particles and plankton of coastal marine ecosystem, which further suggests that viral genomic information may be transported between biomes.

  15. Genetic and biological variation among nucleopolyhedrovirus isolates from the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae).

    PubMed

    Rowley, Daniel L; Farrar, Robert R; Blackburn, Michael B; Harrison, Robert L

    2010-06-01

    A PCR-based method was used to identify and distinguish among 40 uncharacterized nucleopolyhedrovirus (NPV) isolates from larvae of the moth Spodoptera frugiperda that were part of an insect virus collection. Phylogenetic analysis was carried out with sequences amplified from two strongly conserved loci (polh and lef-8) from the 40 isolates in the collection and from eight previously studied S. frugiperda NPV (SfMNPV) isolates. To further distinguish these isolates, analysis was also carried out with sequences from two less-conserved loci, hr4 and hr5. Phylogenetic inference from the sequence data could distinguish among several of the individual isolates and between different groups of isolates from Georgia (USA) and Colombia, South America. A stronger degree of bootstrap support for the phylogenetic trees was obtained with the hr4 and hr5 homologous repeat sequences. Sequencing and phylogenetic analysis detected a relatively high degree of larva-to-larva sequence divergence occurring among isolates of SfMNPV collected from the same field in Missouri, USA. Restriction endonuclease analysis of viral DNA from larvae infected with five isolates from Georgia, Missouri, Louisiana, Florida (USA), and Colombia allowed for further comparison with other previously reported isolates of SfMNPV. Bioassays with these five geographically distinct isolates detected minor differences in virulence. This study highlights the use of PCR to rapidly distinguish and characterize large numbers of historical baculovirus isolates from the same host using minimal quantities of material, and the use of sequences from homologous repeat regions to distinguish closely related isolates of the same NPV species.

  16. Genome sequence of Perigonia lusca single nucleopolyhedrovirus: insights into the evolution of a nucleotide metabolism enzyme in the family Baculoviridae

    PubMed Central

    Ardisson-Araújo, Daniel M. P.; Lima, Rayane Nunes; Melo, Fernando L.; Clem, Rollie J.; Huang, Ning; Báo, Sônia Nair; Sosa-Gómez, Daniel R.; Ribeiro, Bergmann M.

    2016-01-01

    The genome of a novel group II alphabaculovirus, Perigonia lusca single nucleopolyhedrovirus (PeluSNPV), was sequenced and shown to contain 132,831 bp with 145 putative ORFs (open reading frames) of at least 50 amino acids. An interesting feature of this novel genome was the presence of a putative nucleotide metabolism enzyme-encoding gene (pelu112). The pelu112 gene was predicted to encode a fusion of thymidylate kinase (tmk) and dUTP diphosphatase (dut). Phylogenetic analysis indicated that baculoviruses have independently acquired tmk and dut several times during their evolution. Two homologs of the tmk-dut fusion gene were separately introduced into the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) genome, which lacks tmk and dut. The recombinant baculoviruses produced viral DNA, virus progeny, and some viral proteins earlier during in vitro infection and the yields of viral occlusion bodies were increased 2.5-fold when compared to the parental virus. Interestingly, both enzymes appear to retain their active sites, based on separate modeling using previously solved crystal structures. We suggest that the retention of these tmk-dut fusion genes by certain baculoviruses could be related to accelerating virus replication and to protecting the virus genome from deleterious mutation. PMID:27273152

  17. Engineering a Recombinant Baculovirus with a Peptide Hormone Gene and its Effect on the Corn Earworm, Helicoverpa zea

    USDA-ARS?s Scientific Manuscript database

    The helicokinins are peptides identified from Helicoverpa zea that when injected into the larvae were found to cause excessive diuresis and loss of feeding activity. Of the three peptides, helicokinin II (HezK-II) was found to be most potent. A synthetic gene encoding HezK-II was constructed based o...

  18. Effect of emamectin benzoate on mortality, proboscis extension, gustation and reproduction of the corn earworm, Helicoverpa zea

    USDA-ARS?s Scientific Manuscript database

    Newly emerged bollworm adults, Helicoverpa zea (Boddie) require carbohydrate source from plant exudates and nectars for reproduction. Adults actively seek such feeding sites upon eclosion in their natural habitat. We wanted to evaluate this nocturnal behavior of the bollworm for potential use as a p...

  19. Risk assessment for Helicoverpa zea (Lepidoptera: Noctuidae) resistance on dual-gene versus single-gene corn

    USDA-ARS?s Scientific Manuscript database

    Recent changes in EPA regulations have prompted concern in some experts that transgenic corn expressing two lepidopteran-active genes from the soil bacterium Bacillus thuringiensis (Bt) (dual-gene) may result in more rapid selection for resistance in Helicoverpa zea (Boddie) than corn expressing a s...

  20. Complete mitochondrial genome of Helicoverpa zea (Boddie) and expression profiles of mitochondrial-encoded genes in early and late embryos

    USDA-ARS?s Scientific Manuscript database

    The mitochondrial genome of the bollworm, Helicoverpa zea, was assembled using paired-end nucleotide sequence reads generated with a next-generation sequencing platform. Assembly resulted in a mitogenome of 15,348 bp with greater than 17,000-fold average coverage. Organization of the H. zea mitogen...

  1. Prevalence of Helicoverpa zea (Lepidoptera: Noctuidae) on late season volunteer corn in Mississippi: implications on Bt resistance management

    USDA-ARS?s Scientific Manuscript database

    The southern United States has a long growing period between corn harvest and first winter frost, so volunteer corn which germinates after corn harvest has a growing period sufficient for corn earworm, Helicoverpa zea (Boddie) to feed on these plants. However, lower air temperatures can limit larval...

  2. The Host Specificities of Baculovirus per os Infectivity Factors

    PubMed Central

    Song, Jingjiao; Wang, Xi; Hou, Dianhai; Huang, Huachao; Liu, Xijia; Deng, Fei; Wang, Hualin; Arif, Basil M.; Hu, Zhihong; Wang, Manli

    2016-01-01

    Baculoviruses are insect-specific pathogens with a generally narrow host ranges. Successful primary infection is initiated by the proper interaction of at least 8 conserved per os infectivity factors (PIFs) with the host’s midgut cells, a process that remains largely a mystery. In this study, we investigated the host specificities of the four core components of the PIF complex, P74, PIF1, PIF2 and PIF3 by using Helicoverpa armigera nucleopolyhedrovirus (HearNPV) backbone. The four pifs of HearNPV were replaced by their counterparts from a group I Autographa californica multiple nucleopolyhedrovirus (AcMNPV) or a group II Spodoptera litura nucleopolyhedrovirus (SpltNPV). Transfection and infection assays showed that all the recombinant viruses were able to produce infectious budded viruses (BVs) and were lethal to H. armigera larvae via intrahaemocoelic injection. However, feeding experiments using very high concentration of occlusion bodies demonstrated that all the recombinant viruses completely lost oral infectivity except SpltNPV pif3 substituted pif3-null HearNPV (vHaBacΔpif3-Sppif3-ph). Furthermore, bioassay result showed that the median lethal concentration (LC50) value of vHaBacΔpif3-Sppif3-ph was 23-fold higher than that of the control virus vHaBacΔpif3-Hapif3-ph, indicating that SpltNPV pif3 can only partially substitute the function of HearNPV pif3. These results suggested that most of PIFs tested have strict host specificities, which may account, at least in part, for the limited host ranges of baculoviruses. PMID:27454435

  3. Impact of mirid (Creontiades spp.) (Hemiptera: Miridae) pest management on Helicoverpa spp. (Lepidoptera: Noctuidae) outbreaks: the case for conserving natural enemies.

    PubMed

    Knight, Kristen M M; Brier, Hugh B; Lucy, Michael J; Kopittke, Rosemary A

    2007-05-01

    Creontiades spp. (Hemiptera: Miridae) are sucking pests that attack buds, flowers and young pods in mungbeans, Vigna radiata (L.), causing these structures subsequently to abort. If left uncontrolled, mirids can cause 25-50% yield loss. Traditional industry practice has involved prophylactic applications of dimethoate to control mirids at budding and again a week later. The present trial was initiated to highlight the dangers of such a practice, in particular the risk of a subsequent Helicoverpa spp. lepidopteran pest outbreak. A single application of dimethoate halved the population of important natural enemies of Helicoverpa spp., and caused an above-threshold outbreak of Helicoverpa spp. within 11 days. This shows that even a moderate (e.g. 50%) reduction in natural enemies may be sufficient to increase Helicoverpa spp. populations in mungbeans. As a result, prophylactic sprays should not be used for the control of mirids in mungbeans, and dimethoate should be applied only when mirids are above the economic threshold. Indoxacarb was also tested to establish its effect on Helicoverpa spp., mirids and natural enemies. Indoxacarb showed potential for Helicoverpa spp. control and suppression of mirids and had little impact on natural enemies.

  4. The genome sequence of Pseudoplusia includens single nucleopolyhedrovirus and an analysis of p26 gene evolution in the baculoviruses.

    PubMed

    Craveiro, Saluana R; Inglis, Peter W; Togawa, Roberto C; Grynberg, Priscila; Melo, Fernando L; Ribeiro, Zilda Maria A; Ribeiro, Bergmann M; Báo, Sônia N; Castro, Maria Elita B

    2015-02-25

    Pseudoplusia includens single nucleopolyhedrovirus (PsinSNPV-IE) is a baculovirus recently identified in our laboratory, with high pathogenicity to the soybean looper, Chrysodeixis includens (Lepidoptera: Noctuidae) (Walker, 1858). In Brazil, the C. includens caterpillar is an emerging pest and has caused significant losses in soybean and cotton crops. The PsinSNPV genome was determined and the phylogeny of the p26 gene within the family Baculoviridae was investigated. The complete genome of PsinSNPV was sequenced (Roche 454 GS FLX - Titanium platform), annotated and compared with other Alphabaculoviruses, displaying a genome apparently different from other baculoviruses so far sequenced. The circular double-stranded DNA genome is 139,132 bp in length, with a GC content of 39.3 % and contains 141 open reading frames (ORFs). PsinSNPV possesses the 37 conserved baculovirus core genes, 102 genes found in other baculoviruses and 2 unique ORFs. Two baculovirus repeat ORFs (bro) homologs, bro-a (Psin33) and bro-b (Psin69), were identified and compared with Chrysodeixis chalcites nucleopolyhedrovirus (ChchNPV) and Trichoplusia ni single nucleopolyhedrovirus (TnSNPV) bro genes and showed high similarity, suggesting that these genes may be derived from an ancestor common to these viruses. The homologous repeats (hrs) are absent from the PsinSNPV genome, which is also the case in ChchNPV and TnSNPV. Two p26 gene homologs (p26a and p26b) were found in the PsinSNPV genome. P26 is thought to be required for optimal virion occlusion in the occlusion bodies (OBs), but its function is not well characterized. The P26 phylogenetic tree suggests that this gene was obtained from three independent acquisition events within the Baculoviridae family. The presence of a signal peptide only in the PsinSNPV p26a/ORF-20 homolog indicates distinct function between the two P26 proteins. PsinSNPV has a genomic sequence apparently different from other baculoviruses sequenced so far. The complete

  5. Field efficacy and transmission of fast- and slow-killing nucleopolyhedroviruses that are infectious to Adoxophyes honmai (Lepidoptera: Tortricidae).

    PubMed

    Takahashi, Maho; Nakai, Madoka; Saito, Yasumasa; Sato, Yasushi; Ishijima, Chikara; Kunimi, Yasuhisa

    2015-03-18

    The smaller tea tortrix, Adoxophyes honmai (Lepidoptera: Tortricidae), is an economically important pest of tea in Japan. Previous work showed that a fast-killing nucleopolyhedrovirus (NPV) isolated from A. orana (AdorNPV) and a slow-killing NPV isolated from A. honmai (AdhoNPV) are both infectious to A. honmai larvae. Field application of these different NPVs was conducted against an A. honmai larval population in tea plants, and the control efficacy and transmission rate of the two NPVs were compared. The slow-killing AdhoNPV showed lower field efficacy, in terms of preventing damage caused by A. honmai larvae against the tea plants, than the fast-killing AdorNPV. However, AdhoNPV had a significantly higher horizontal transmission rate than AdorNPV. These results show that AdorNPV is suitable as an inundative agent, while AdhoNPV is an appropriate inoculative agent.

  6. The putative LEF-1 proteins from two distinct Choristoneura fumiferana multiple nucleopolyhedroviruses share domain homology to eukaryotic primases.

    PubMed

    Barrett, J W; Lauzon, H A; Mercuri, P S; Krell, P J; Sohi, S S; Arif, B M

    1996-01-01

    We have identified the lef-1 genes from two multiple nucleopolyhedroviruses that infect natural populations of Choristoneura fumiferana. The lef-1 genes in both viruses are directly upstream and in the opposite orientation of their respective ecdysteroid UDP-glucosyltransferase (egt) genes. This gene organization pattern is similar to that found in the genomes of AcMNPV and of OpMNPV. As well, the coding regions and putative protein sequences share a high degree of similarity. Alignment of the predicted amino acid sequences of all known baculovirus lef-1 genes suggests that the LEF-1 proteins have a relatively high degree of conservation, particularly at four identified and distinct domains. Moreover, LEF-1 proteins bear clear similarity to some eukaryotic primases, predominately at three of the four domains where certain amino acids are absolutely conserved.

  7. The BRO proteins of Bombyx mori nucleopolyhedrovirus are nucleocytoplasmic shuttling proteins that utilize the CRM1-mediated nuclear export pathway

    SciTech Connect

    Kang, Won Kyung . E-mail: wkkang@riken.jp; Kurihara, Masaaki . E-mail: mkuri@riken.jp; Matsumoto, Shogo . E-mail: smatsu@riken.jp

    2006-06-20

    The BRO proteins of Bombyx mori nucleopolyhedrovirus (BmNPV) display a biphasic pattern of intracellular localization during infection. At early times, they reside in the nucleus but then show both cytoplasmic and nuclear localization as the infection proceeds. Therefore, we examined the possibility of nuclear export. Using inhibitors, we reveal that BmNPV BRO proteins shuttle between the nucleus and cytoplasm. Mutations on the leucine-rich region of BRO proteins resulted in nuclear accumulation of transiently expressed proteins, suggesting that this region functions as a CRM1-dependent nuclear export signal (NES). On the contrary, mutant BRO-D with an altered NES did not show nuclear accumulation in infected cells, although protein production seemed to be reduced. RT-PCR analysis showed that the lower level of protein production was due to a reduction in RNA synthesis. Taken together, our results suggest that BRO proteins are nucleocytoplasmic shuttling proteins that utilize the CRM1-mediated nuclear export pathway.

  8. The BRO proteins of Bombyx mori nucleopolyhedrovirus are nucleocytoplasmic shuttling proteins that utilize the CRM1-mediated nuclear export pathway.

    PubMed

    Kang, WonKyung; Kurihara, Masaaki; Matsumoto, Shogo

    2006-06-20

    The BRO proteins of Bombyx mori nucleopolyhedrovirus (BmNPV) display a biphasic pattern of intracellular localization during infection. At early times, they reside in the nucleus but then show both cytoplasmic and nuclear localization as the infection proceeds. Therefore, we examined the possibility of nuclear export. Using inhibitors, we reveal that BmNPV BRO proteins shuttle between the nucleus and cytoplasm. Mutations on the leucine-rich region of BRO proteins resulted in nuclear accumulation of transiently expressed proteins, suggesting that this region functions as a CRM1-dependent nuclear export signal (NES). On the contrary, mutant BRO-D with an altered NES did not show nuclear accumulation in infected cells, although protein production seemed to be reduced. RT-PCR analysis showed that the lower level of protein production was due to a reduction in RNA synthesis. Taken together, our results suggest that BRO proteins are nucleocytoplasmic shuttling proteins that utilize the CRM1-mediated nuclear export pathway.

  9. A novel third chromosomal locus controls susceptibility to Autographa californica multiple nucleopolyhedrovirus in the silkworm, Bombyx mori.

    PubMed

    Xu, Jian; Kusakabe, Takahiro; Yamamoto, Kimiko; Suetsugu, Yoshitaka; Mon, Hiroaki; Li, Zhiqing; Zhu, Li; Iiyama, Kazuhiro; Banno, Yutaka; Yoshimura, Kaito; Lee, Jae Man

    2014-04-01

    Baculovirus demonstrates specific infection spectrums and thus one certain host exhibits particular response to single baculovirus isolate. Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is considered to be not an innate pathogen to Bombyx mori, but some silkworm strains have been identified to be permissive to AcMNPV, indicating the positive or negative involvement of certain host factors in baculovirus replications in vivo. To provide a fundamental knowledge of this process, we performed large-scale screening to investigate the responses of 448 silkworm strains against recombinant AcMNPV inoculation. By genetic analysis between permissive and resistant strains identified, we further confirmed that a potential corresponding locus on chromosome 3 regulates host responses to AcMNPV in silkworm. Additionally, we found that it is available for AcMNPV-silkworm baculovirus expression vector system to produce proteins of interest.

  10. Genomic comparison of Neodiprion sertifer and Neodiprion lecontei nucleopolyhedroviruses and identification of potential hymenopteran baculovirus-specific open reading frames.

    PubMed

    Lauzon, Hilary A M; Garcia-Maruniak, Alejandra; Zanotto, Paolo M de A; Clemente, José C; Herniou, Elisabeth A; Lucarotti, Christopher J; Arif, Basil M; Maruniak, James E

    2006-06-01

    Genomic comparison of Neodiprion sertifer nucleopolyhedrovirus (NeseNPV) and Neodiprion lecontei nucleopolyhedrovirus (NeleNPV) showed that the hymenopteran baculoviruses had features in common and were distinct from other, fully sequenced lepidopteran and dipteran baculoviruses. Their genomes were small in size (86,462 and 81,755 bp, respectively), had low G+C contents (33.8 and 33.3 mol%, respectively) and contained fewer open reading frames (ORFs) (90 and 89, respectively) than other baculoviruses. They shared 69 ORFs (48.6% mean amino acid identity overall), 43 of which were previously identified baculovirus homologues. The remaining shared ORFs could be common to other baculoviruses, but low amino acid identities precluded identifying them as such. Some may also be unique to hymenopteran baculoviruses. These included a trypsin-like protease, a zinc-finger protein, regulator of chromosome condensation proteins, a densovirus capsid-like protein and a phosphotransferase. Structural analysis, the presence of conserved domains and phylogenetic studies suggested that some of these ORFs may be functional and could have been transferred horizontally from an insect host. ORFs found only in NeseNPV and NeleNPV may play a role in host specificity and/or tissue tropism, as hymenopteran baculoviruses are restricted to the midgut. The genomes were basically collinear, but contained non-syntenic regions (NSRs) with large numbers of repeats between their polyhedrin and dbp genes. They differed from each other in the number of ORFs and the G+C content of their NSRs and the presence of homologous regions in the NeseNPV genome. NeleNPV also had a short inversion relative to NeseNPV. NeseNPV contained 21 ORFs not found in NeleNPV and NeleNPV had 20 ORFs not found in NeseNPV.

  11. Lacanobia oleracea nucleopolyhedrovirus (LaolNPV): A new European species of alphabaculovirus with a narrow host range

    PubMed Central

    Erlandson, Martin A.; Frayssinet, Marie; Williams, Trevor; Theilmann, David A.; Volkoff, Anne-Nathalie; Caballero, Primitivo

    2017-01-01

    During an insect sampling program in alfalfa crops near Montpellier, France in 2011, Lacanobia oleracea larvae were collected that died due to nucleopolyhedrovirus infection (LaolNPV). This virus was subjected to molecular and biological characterization. The virus was a multiple nucleocapsid NPV that showed similar restriction profiles to Mamestra configurata NPV-A (MacoNPV-A) but with significant differences. Polypeptide analysis demonstrated similar proteins in occlusion bodies and occlusion derived virions, to those observed in NPVs from Mamestra spp. Terminal sequencing revealed that the genome organization shared similarity with that of MacoNPV-A. The most homologous virus was MacoNPV-A 90/2 isolate (95.63% identity and 96.47% similarity), followed by MacoNPV-A 90/4 strain (95.37% and 96.26%), MacoNPV-B (89.21% and 93.53%) and M. brassicae MNPV (89.42% and 93.74%). Phylogenetic analysis performed with lef-8, lef-9, polh and a concatenated set of genes showed that LaolNPV and the Mamestra spp. NPVs clustered together with HaMNPV, but with a closer genetic distance to MacoNPV-A strains. The Kimura 2-parameter (K-2-P) distances of the complete genes were greater than 0.05 between LaolNPV and the MbMNPV/MacoNPV-B/HaMNPV complex, which indicates that LaolNPV is a distinct species. K-2-P distances were in the range 0.015–0.050 for comparisons of LaolNPV with MacoNPV-A strains, such that additional biological characteristics should be evaluated to determine species status. While MacoNPV-A was pathogenic to seven lepidopteran species tested, LaolNPV was only pathogenic to Chrysodeixis chalcites. Given these findings, Lacanobia oleracea nucleopolyhedrovirus should be considered as a new species in the Alphabaculovirus genus. PMID:28426736

  12. Expression of the lef5 gene from Spodoptera exigua multiple nucleopolyhedrovirus contributes to the baculovirus stability in cell culture.

    PubMed

    Martínez-Solís, María; Jakubowska, Agata K; Herrero, Salvador

    2017-09-09

    Baculoviruses are a broad group of viruses infecting insects, predominately of the order Lepidoptera. They are used worldwide as biological insecticides and as expression vectors to produce recombinant proteins. Baculoviruses replicate in their host, although several cell lines have been developed for in vitro replication. Nevertheless, replication of baculoviruses in cell culture involves the generation of defective viruses with a decrease in productivity and virulence. Transcriptional studies of the Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) and the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infective process revealed differences in the expression patterns when the virus replicated under in vitro (Se301 cells) or in vivo (S. exigua larvae) conditions. The late expression factor 5 (lef5) gene was found to be highly overexpressed when the virus replicates in larvae. To test the possible role of lef5 expression in viral stability, recombinant AcMNPV expressing the lef5 gene from SeMNPV (Se-lef5) was generated and its stability was monitored during successive infection passages in Sf21 cells by evaluating the loss of several essential and non-essential genes. The gfp transgene was more stable in those viruses expressing the Se-LEF5 protein and the GFP-defective viruses were accumulated at a lower level when compared to its control viruses, confirming the positive influence of lef5 in viral stability during the multiplication process. This work describes for the first time a viral factor involved in transgene stability when baculoviruses replicate in cell culture, opening new ways to facilitate the in vitro production of recombinant proteins using baculovirus.

  13. Lacanobia oleracea nucleopolyhedrovirus (LaolNPV): A new European species of alphabaculovirus with a narrow host range.

    PubMed

    Simón, Oihane; Erlandson, Martin A; Frayssinet, Marie; Williams, Trevor; Theilmann, David A; Volkoff, Anne-Nathalie; Caballero, Primitivo

    2017-01-01

    During an insect sampling program in alfalfa crops near Montpellier, France in 2011, Lacanobia oleracea larvae were collected that died due to nucleopolyhedrovirus infection (LaolNPV). This virus was subjected to molecular and biological characterization. The virus was a multiple nucleocapsid NPV that showed similar restriction profiles to Mamestra configurata NPV-A (MacoNPV-A) but with significant differences. Polypeptide analysis demonstrated similar proteins in occlusion bodies and occlusion derived virions, to those observed in NPVs from Mamestra spp. Terminal sequencing revealed that the genome organization shared similarity with that of MacoNPV-A. The most homologous virus was MacoNPV-A 90/2 isolate (95.63% identity and 96.47% similarity), followed by MacoNPV-A 90/4 strain (95.37% and 96.26%), MacoNPV-B (89.21% and 93.53%) and M. brassicae MNPV (89.42% and 93.74%). Phylogenetic analysis performed with lef-8, lef-9, polh and a concatenated set of genes showed that LaolNPV and the Mamestra spp. NPVs clustered together with HaMNPV, but with a closer genetic distance to MacoNPV-A strains. The Kimura 2-parameter (K-2-P) distances of the complete genes were greater than 0.05 between LaolNPV and the MbMNPV/MacoNPV-B/HaMNPV complex, which indicates that LaolNPV is a distinct species. K-2-P distances were in the range 0.015-0.050 for comparisons of LaolNPV with MacoNPV-A strains, such that additional biological characteristics should be evaluated to determine species status. While MacoNPV-A was pathogenic to seven lepidopteran species tested, LaolNPV was only pathogenic to Chrysodeixis chalcites. Given these findings, Lacanobia oleracea nucleopolyhedrovirus should be considered as a new species in the Alphabaculovirus genus.

  14. Genetic and Biological Characterization of Four Nucleopolyhedrovirus Isolates Collected in Mexico for the Control of Spodoptera exigua (Lepidoptera: Noctuidae).

    PubMed

    Zamora-Avilés, N; Murillo, R; Lasa, R; Pineda, S; Figueroa, J I; Bravo-Patiño, A; Díaz, O; Corrales, J L; Martínez, A M

    2017-08-01

    This study describes four multiple nucleocapsid nucleopolyhedrovirus isolates recovered from infected larvae of beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), on crops in two different geographical regions of Mexico. Molecular and biological characterization was compared with characterized S. exigua multiple nucleopolyhedrovirus (SeMNPV) isolates from the United States (SeUS1 and SeUS2) and Spain (SeSP2). Restriction endonuclease analysis of viral DNA confirmed that all Mexican isolates were SeMNPV isolates, but molecular differences between the Mexican and the reference isolates were detected using PCR combined with restriction fragment length polymorphism (RFLP). Amplification of the variable region V01 combined with RFLP distinguished the two Mexican isolates, SeSLP6 and SeSIN6. BglII digestions showed that the majority of the isolates contained submolar bands, indicating the presence of genetic heterogeneity. Amplification of the variable regions V04 and V05 distinguished between American and the Spanish isolates. Biological characterization was performed against two laboratory colonies of S. exigua, one from Mexico, and another from Switzerland. Insects from the Mexican colony were less susceptible to infection than insects from Se-Swiss colony. In the Se-Mex colony, SeSP2 was the most pathogenic isolate followed by SeSIN6, although their virulence was similar to most of the isolates tested. In Se-Swiss colony, similar LD50 values were observed for the five isolates, although the virulence was higher for the SeSLP6 isolate, which also had the highest OB (occlusion body) yield. We conclude that the Mexican isolates SeSIN6 and SeSLP6 possess insecticidal traits of value for the development of biopesticides for the control of populations of S. exigua. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  15. Efficient isolation, purification, and characterization of the Helicoverpa zea VHDL receptor.

    PubMed

    Persaud, Deryck R; Yousefi, Vandad; Haunerland, Norbert

    2003-12-01

    The study of fat body receptors (e.g., VHDL receptor) in Lepidoptera has been irksome due to the fact that isolation and purification of these proteins are difficult and resulted in extremely low yields. A rapid and efficient method is presented for the purification of Helicoverpa zea VHDL receptor by the use of VHDL-biotin ligand complexed to streptavidin coated magnetic beads. The technique can be easily applied to other ligands and allows for the purification of membrane proteins with higher yields compared to previously used methods involving immunopurification. Although the purified protein can be characterized by Western and non-radioactive ligand blots using enhanced chemiluminescence (ECL), a non-radioactive ligand blot method using VHDL-FITC is presented, which allows for the quick analysis of the receptor directly from the blot under standard UV light. Sufficient receptor protein has been derived for amino acid analysis, receptor-ligand and xenobiotic binding studies.

  16. Storage stability of Anagrapha falcifera nucleopolyhedrovirus in spray-dried formulations.

    PubMed

    Tamez-Guerra, Patricia; McGuire, Michael R; Behle, Robert W; Shasha, Baruch S; Pingel, Randall L

    2002-01-01

    A multiply embedded nucleopolyhedrovirus isolated from Anagrapha falcifera (Kirby) (AfMNPV) can lose insecticidal activity during months of dry storage in ambient room conditions. We tested the spray-dried AfMNPV formulations after storage for up to 1 year at room temperatures for insecticidal activity against neonate Trichoplusia ni (Hübner). Experimental formulations were made using combinations of corn flours, lignin, and sucrose, and were selected based on previous work which demonstrated that these formulations resisted solar degradation in field experiments. Twelve experimental formulations (organized in three groups of four formulations) compared the effect of (1) the ratio of formulation ingredients (lignin and corn flour) to virus concentration, (2) different sources of lignin, or (3) different corn flours and sugar. Based on a single-dose plant assay with these 12 formulations, none of the formulations lost significant activity due to the drying process, when compared with the unformulated wet AfMNPV. Samples of the 12 dried formulations were stored at room (22+/-3 degrees C) and refrigerated (4 degrees C) temperatures. Insecticidal activity (LC(50)) was determined with a dosage-response assay for neonates fed on treated cotton-leaf disks. After 6 (or 9) and 12 months storage, refrigerated samples maintained insecticidal activity better than corresponding samples stored at room temperatures with LC(50)s that averaged 2.0 x 10(6) polyhedral inclusion bodies per milliliter (pibs/ml) for refrigerated samples and 5.4 x 10(6) pibs/ml for samples stored at room temperatures. Compared with unformulated stock virus stored frozen, six formulations stored at room temperature and 10 formulations stored in the refrigerator did not lose significant insecticidal activity after 1 year based on overlapping 90% confidence intervals. Changing the ratio of virus to formulation ingredients did not provide a clear trend over the range of concentrations tested, and may be

  17. Autographa californica multiple nucleopolyhedrovirus ODV-E56 envelope protein is required for oral infectivity and can be substituted functionally by Rachiplusia ou multiple nucleopolyhedrovirus ODV-E56.

    PubMed

    Harrison, Robert L; Sparks, Wendy O; Bonning, Bryony C

    2010-05-01

    The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) odv-e56 gene encodes an occlusion-derived virus (ODV)-specific envelope protein, ODV-E56. In a previous analysis, the odv-e56 gene was found to be under positive selection pressure, suggesting that it may be a determinant of virus host range. To assess the role of ODV-E56 in oral infectivity and host range, we constructed recombinant AcMNPV clones (Ac69GFP-e56lacZ and AcIEGFP-e56lacZ) in which ODV-E56 protein synthesis was eliminated by inserting a beta-galactosidase (lacZ) expression cassette into the odv-e56 open reading frame. We also constructed a recombinant virus, Ac69GFP-Roe56, in which the native AcMNPV odv-e56 coding sequence was replaced with that of Rachiplusia ou multiple nucleopolyhedrovirus (RoMNPV), a closely related virus that is significantly more virulent towards some host species than AcMNPV. The odv-e56 recombinant viruses exhibited no alterations in polyhedron production and morphogenesis or in the production of infectious budded virus in cell culture. In bioassays using three lepidopteran host species, the oral infectivities of the odv-e56 mutant viruses Ac69GFP-e56lacZ and AcIEGFP-e56lacZ were profoundly impaired compared with those of wild-type and control recombinant viruses. Oral infectivity was restored fully by marker rescue of the odv-e56 mutant viruses with either the AcMNPV or the RoMNPV odv-e56 gene. In bioassays using two host species that are more susceptible to RoMNPV than to AcMNPV, Ac69GFP-Roe56 killed larvae with LC50 values similar to those of recombinant viruses expressing AcMNPV ODV-E56. This result indicated that replacement of the AcMNPV odv-e56 gene with the RoMNPV orthologue did not increase virulence against these two species.

  18. Trichoplusia ni Kinesin-1 Associates with Autographa californica Multiple Nucleopolyhedrovirus Nucleocapsid Proteins and Is Required for Production of Budded Virus

    PubMed Central

    Biswas, Siddhartha; Blissard, Gary W.

    2016-01-01

    ABSTRACT The mechanism by which nucleocapsids of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) egress from the nucleus to the plasma membrane, leading to the formation of budded virus (BV), is not known. AC141 is a nucleocapsid-associated protein required for BV egress and has previously been shown to be associated with β-tubulin. In addition, AC141 and VP39 were previously shown by fluorescence resonance energy transfer by fluorescence lifetime imaging to interact directly with the Drosophila melanogaster kinesin-1 light chain (KLC) tetratricopeptide repeat (TPR) domain. These results suggested that microtubule transport systems may be involved in baculovirus nucleocapsid egress and BV formation. In this study, we investigated the role of lepidopteran microtubule transport using coimmunoprecipitation, colocalization, yeast two-hybrid, and small interfering RNA (siRNA) analyses. We show that nucleocapsid AC141 associates with the lepidopteran Trichoplusia ni KLC and kinesin-1 heavy chain (KHC) by coimmunoprecipitation and colocalization. Kinesin-1, AC141, and microtubules colocalized predominantly at the plasma membrane. In addition, the nucleocapsid proteins VP39, FP25, and BV/ODV-C42 were also coimmunoprecipitated with T. ni KLC. Direct analysis of the role of T. ni kinesin-1 by downregulation of KLC by siRNA resulted in a significant decrease in BV production. Nucleocapsids labeled with VP39 fused with three copies of the mCherry fluorescent protein also colocalized with microtubules. Yeast two-hybrid analysis showed no evidence of a direct interaction between kinesin-1 and AC141 or VP39, suggesting that either other nucleocapsid proteins or adaptor proteins may be required. These results further support the conclusion that microtubule transport is required for AcMNPV BV formation. IMPORTANCE In two key processes of the replication cycle of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV), nucleocapsids are

  19. Development of a Microbial-Based Integrated Pest Management Program for Helicoverpa spp. (Lepidoptera: Noctuidae) and Beneficial Insects on Conventional Cotton Crops in Australia

    PubMed Central

    Mensah, Robert K.; Young, Alison; Rood-England, Leah

    2015-01-01

    Entomopathogenic fungi, when used as a microbial control agent against cotton pests, such as Helicoverpa spp., may have the potential to establish and spread in the environment and to have an impact on both pests and beneficial insects. Information on the effect of entomopathogenic fungi on pests and beneficial insects is crucial for a product to be registered as a biopesticide. The effect of the entomopathogenic fungus BC 639 (Aspergillus sp.) against Helicoverpa spp. and beneficial insects (mostly predatory insects) was studied in the laboratory and in cotton field trials. The results show that when Helicoverpa spp. second instar larvae were exposed to increasing concentrations (from 102 to 109) of the entomopathogenic fungus BC 639, the optimum dose required to kill over 50% of the insects was 1.0 × 107 spores/mL. In the field trials, the number of Helicoverpa spp. per metre on plots treated with 1.0 or 0.50 L/ha of BC 639 was the same as on plots treated with the recommended rate of the commercial insecticide, Indoxacarb. However, when plots were treated with 0.25 L/ha of BC 639, this was not as effective at controlling Helicoverpa spp. as 1.0 or 0.5 L/ha BC 639 or Indoxacarb. BC 639 had less effect on predatory insects when applied at lower rates (0.50 and 0.25 L/ha) than at higher rates (1.0 L/ha). Thus, BC 639 was more selective against predators when applied at lower rates than at the higher rate, but was also more selective than Indoxacarb. Thus, the ability of BC 639 to control Helicoverpa spp. effectively with a minimal effect on predatory insects indicates its potential for enhancing integrated pest management programs and to sustain cotton production. PMID:26463189

  20. Development of a Microbial-Based Integrated Pest Management Program for Helicoverpa spp. (Lepidoptera: Noctuidae) and Beneficial Insects on Conventional Cotton Crops in Australia.

    PubMed

    Mensah, Robert K; Young, Alison; Rood-England, Leah

    2015-04-09

    Entomopathogenic fungi, when used as a microbial control agent against cotton pests, such as Helicoverpa spp., may have the potential to establish and spread in the environment and to have an impact on both pests and beneficial insects. Information on the effect of entomopathogenic fungi on pests and beneficial insects is crucial for a product to be registered as a biopesticide. The effect of the entomopathogenic fungus BC 639 (Aspergillus sp.) against Helicoverpa spp. and beneficial insects (mostly predatory insects) was studied in the laboratory and in cotton field trials. The results show that when Helicoverpa spp. second instar larvae were exposed to increasing concentrations (from 10² to 10⁸) of the entomopathogenic fungus BC 639, the optimum dose required to kill over 50% of the insects was 1.0 ´ 10⁷ spores/mL. In the field trials, the number of Helicoverpa spp. per metre on plots treated with 1.0 or 0.50 L/ha of BC 639 was the same as on plots treated with the recommended rate of the commercial insecticide, Indoxacarb. However, when plots were treated with 0.25 L/ha of BC 639, this was not as effective at controlling Helicoverpa spp. as 1.0 or 0.5 L/ha BC 639 or Indoxacarb. BC 639 had less effect on predatory insects when applied at lower rates (0.50 and 0.25 L/ha) than at higher rates (1.0 L/ha). Thus, BC 639 was more selective against predators when applied at lower rates than at the higher rate, but was also more selective than Indoxacarb. Thus, the ability of BC 639 to control Helicoverpa spp. effectively with a minimal effect on predatory insects indicates its potential for enhancing integrated pest management programs and to sustain cotton production.

  1. Susceptibilities of Helicoverpa zea and Heliothis virescens (Lepidoptera: Noctuidae) populations to Cry1Ac insecticidal protein.

    PubMed

    Ali, M I; Luttrell, R G; Young, S Y

    2006-02-01

    Susceptibilities of bollworm, Helicoverpa zea (Boddie) and tobacco budworm, Heliothis virescens (F.) to Cry1Ac were measured via a diet-incorporated assay with MPV II at the University of Arkansas during 2002-2004. Lethal concentration-mortality (LC50) estimates of five laboratory, seven laboratory-cross, and 10 field populations of H. virescens varied 12-fold. Pooled susceptibilities of H. virescens across all laboratory and field populations varied five-fold. The LC50 estimates for H. virescens were higher than those reported by previous research before the introduction of transgenic crops. However, the ratio of susceptibility of laboratory and field populations was similar, suggesting no change in overall species susceptibility. Individual LC50 estimates of five laboratory, nine laboratory-cross, and 57 field populations of H. zea varied over 130-fold. Pooled susceptibilities across laboratory and field populations varied widely. Among the field populations, colonies from non-Bacillus thuringiensis (Bt) crops were generally more susceptible than those from Bt crops. Across the Bt crops expressing Cry protein, colonies from Bollgard (Monsanto Company) cotton had lower susceptibility to CrylAc than those from Bt corn and those from non-Bt crops.

  2. Incipient Resistance of Helicoverpa punctigera to the Cry2Ab Bt Toxin in Bollgard II® Cotton

    PubMed Central

    Downes, Sharon; Parker, Tracey; Mahon, Rod

    2010-01-01

    Combinations of dissimilar insecticidal proteins (“pyramids”) within transgenic plants are predicted to delay the evolution of pest resistance for significantly longer than crops expressing a single transgene. Field-evolved resistance to Bacillus thuringiensis (Bt) transgenic crops has been reported for first generation, single-toxin varieties and the Cry1 class of proteins. Our five year data set shows a significant exponential increase in the frequency of alleles conferring Cry2Ab resistance in Australian field populations of Helicoverpa punctigera since the adoption of a second generation, two-toxin Bt cotton expressing this insecticidal protein. Furthermore, the frequency of cry2Ab resistance alleles in populations from cropping areas is 8-fold higher than that found for populations from non-cropping regions. This report of field evolved resistance to a protein in a dual-toxin Bt-crop has precisely fulfilled the intended function of monitoring for resistance; namely, to provide an early warning of increases in frequencies that may lead to potential failures of the transgenic technology. Furthermore, it demonstrates that pyramids are not ‘bullet proof’ and that rapid evolution to Bt toxins in the Cry2 class is possible. PMID:20830203

  3. Evolution of Resistance by Helicoverpa zea (Lepidoptera: Noctuidae) Infesting Insecticidal Crops in the Southern United States

    PubMed Central

    Onstad, David; Crain, Philip; Crespo, Andre; Hutchison, William; Buntin, David; Porter, Pat; Catchot, Angus; Cook, Don; Pilcher, Clint; Flexner, Lindsey; Higgins, Laura

    2016-01-01

    We created a deterministic, frequency-based model of the evolution of resistance by corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), to insecticidal traits expressed in crops planted in the heterogeneous landscapes of the southern United States. The model accounts for four generations of selection by insecticidal traits each year. We used the model results to investigate the influence of three factors on insect resistance management (IRM): 1) how does adding a third insecticidal trait to both corn and cotton affect durability of the products, 2) how does unstructured corn refuge influence IRM, and 3) how do block refuges (50% compliance) and blended refuges compare with regard to IRM? When Bt cotton expresses the same number of insecticidal traits, Bt corn with three insecticidal traits provides longer durability than Bt corn with two pyramided traits. Blended refuge provides similar durability for corn products compared with the same level of required block refuge when the rate of refuge compliance by farmers is 50%. Results for Mississippi and Texas are similar, but durabilities for corn traits are surprisingly lower in Georgia, where unstructured corn refuge is the highest of the three states, but refuge for Bt cotton is the lowest of the three states. Thus, unstructured corn refuge can be valuable for IRM but its influence is determined by selection for resistance by Bt cotton. PMID:26637533

  4. Evolution of Resistance by Helicoverpa zea (Lepidoptera: Noctuidae) Infesting Insecticidal Crops in the Southern United States.

    PubMed

    Pan, Zaiqi; Onstad, David; Crain, Philip; Crespo, Andre; Hutchison, William; Buntin, David; Porter, Pat; Catchot, Angus; Cook, Don; Pilcher, Clint; Flexner, Lindsey; Higgins, Laura

    2016-04-01

    We created a deterministic, frequency-based model of the evolution of resistance by corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), to insecticidal traits expressed in crops planted in the heterogeneous landscapes of the southern United States. The model accounts for four generations of selection by insecticidal traits each year. We used the model results to investigate the influence of three factors on insect resistance management (IRM): 1) how does adding a third insecticidal trait to both corn and cotton affect durability of the products, 2) how does unstructured corn refuge influence IRM, and 3) how do block refuges (50% compliance) and blended refuges compare with regard to IRM? When Bt cotton expresses the same number of insecticidal traits, Bt corn with three insecticidal traits provides longer durability than Bt corn with two pyramided traits. Blended refuge provides similar durability for corn products compared with the same level of required block refuge when the rate of refuge compliance by farmers is 50%. Results for Mississippi and Texas are similar, but durabilities for corn traits are surprisingly lower in Georgia, where unstructured corn refuge is the highest of the three states, but refuge for Bt cotton is the lowest of the three states. Thus, unstructured corn refuge can be valuable for IRM but its influence is determined by selection for resistance by Bt cotton.

  5. Aflatoxin B1 detoxification by CYP321A1 in Helicoverpa zea.

    PubMed

    Niu, Guodong; Wen, Zhimou; Rupasinghe, Sanjeewa G; Zeng, Ren Sen; Berenbaum, May R; Schuler, Mary A

    2008-09-01

    The polyphagous corn earworm Helicoverpa zea frequently encounters aflatoxins, mycotoxins produced by the pathogens Aspergillus flavus and A. parasiticus, which infect many of this herbivore's host plants. While aflatoxin B1 metabolism by midgut enzymes isolated from fifth instars feeding on control diets was not detected, this compound was metabolized by midgut enzymes isolated from larvae consuming diets supplemented with xanthotoxin, coumarin, or indole-3-carbinol, phytochemicals that are likely to co-occur with aflatoxin in infected host plants. Of the two metabolites generated, the main derivative identified in midguts induced with these chemicals and in reactions containing heterologously expressed CYP321A1 was aflatoxin P1 (AFP1), an O-demethylated product of AFB1. RT-PCR gel blots indicated that the magnitude of CYP321A1 transcript induction by these chemicals is associated with the magnitude of increase in the metabolic activities of induced midgut enzymes (coumarin>xanthotoxin>indole 3-carbinol). These results indicate that induction of P450s, such as CYP321A1, plays an important role in reducing AFB1 toxicity to H. zea. Docking of AFB1 in the molecular models of CYP321A1 and CYP6B8 highlights differences in their proximal catalytic site volumes that allow only CYP321A1 to generate the AFP1 metabolite. Copyright 2008 Wiley-Liss, Inc.

  6. Larval Helicoverpa zea Transcriptional, Growth and Behavioral Responses to Nicotine and Nicotiana tabacum

    PubMed Central

    Gog, Linus; Vogel, Heiko; Hum-Musser, Sue M.; Tuter, Jason; Musser, Richard O.

    2014-01-01

    The polyphagous feeding habits of the corn earworm, Helicoverpa zea (Boddie), underscore its status as a major agricultural pest with a wide geographic distribution and host plant repertoire. To study the transcriptomic response to toxins in diet, we conducted a microarray analysis of H. zea caterpillars feeding on artificial diet, diet laced with nicotine and Nicotiana tabacum (L.) plants. We supplemented our analysis with growth and aversion bioassays. The transcriptome reflects an abundant expression of proteases, chitin, cytochrome P450 and immune-related genes, many of which are shared between the two experimental treatments. However, the tobacco treatment tended to elicit stronger transcriptional responses than nicotine-laced diet. The salivary factor glucose oxidase, known to suppress nicotine induction in the plant, was upregulated by H. zea in response to tobacco but not to nicotine-laced diet. Reduced caterpillar growth rates accompanied the broad regulation of genes associated with growth, such as juvenile hormone epoxide hydrolase. The differential expression of chemosensory proteins, such as odorant binding-protein-2 precursor, as well as the neurotransmitter nicotinic-acetylcholine-receptor subunit 9, highlights candidate genes regulating aversive behavior towards nicotine. We suggest that an observed coincidental rise in cannibalistic behavior and regulation of proteases and protease inhibitors in H. zea larvae signify a compensatory response to induced plant defenses. PMID:26462833

  7. Enzymatic decomposition of elicitors of plant volatiles in Heliothis virescens and Helicoverpa zea.

    PubMed

    Mori, N; Alborn, H T.; Teal, P E.A.; Tumlinson, J H.

    2001-07-01

    Feeding by larvae of Heliothis virescens induces cotton, corn and tobacco plants to release blends of volatile organic compounds that differ in constituent proportions from blends released when Helicoverpa zea larvae feed on the same plant species. The same elicitors (and analogs) of plant biosynthesis and release of volatiles, originally identified in oral secretions of Spodoptera exigua larvae, were also found in oral secretions of H. virescens and H. zea. However, relative amounts of these compounds, particularly N-(17-hydroxylinolenoyl)-L-glutamine (volicitin), 17-hydroxylinolenic acid, and N-linolenoyl-L-glutamine, varied among batches of oral secretions, more so in H. virescens than in H. zea. This variation was due to cleavage of the amide bond of the fatty acid-amino acid conjugates by an enzyme, or enzymes, originating in the midgut. The enzymatic activity in guts of H. virescens was significantly greater than that found in guts of H. zea. Furthermore, H. zea frass contains N-linolenoyl-L-glutamine in more than 0.1% wet weight, while this conjugate comprises