Cloning and comparative characterization of nucleopolyhedroviruses isolated from African Bollworm, Helicoverpa armigera, (Lepidoptera: Noctudiae) in different geographic regions

  • Gordon Ogembo Javier
    Laboratory of Biodynamics, Graduate School of Bioagricultural Sciences Nagoya University
  • Chaeychomsri Sudawan
    Central Laboratory and Greenhouse Complex, Kasetsart University
  • Kamiya Katsumi
    Gifu Prefectural Institute for Bioengineering
  • Ishikawa Hiroki
    Laboratory of Biodynamics, Graduate School of Bioagricultural Sciences Nagoya University
  • Katou Yasuhiro
    Laboratory of Biodynamics, Graduate School of Bioagricultural Sciences Nagoya University Division of Biomodeling, Graduate School of Bioagricultural Sciences, Nagoya University
  • Ikeda Motoko
    Laboratory of Sericulture and Entomoresources, Graduate School of Bioagricultural Sciences Nagoya University
  • Kobayashi Michihiro
    Laboratory of Biodynamics, Graduate School of Bioagricultural Sciences Nagoya University

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タイトル別名
  • Cloning and Comparative Characterization of Nucleopolyhedroviruses Isolated from African Bollworm, <i>Helicoverpa armigera</i>,(Lepidoptera: Noctudiae) in Different Geographic Regions

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A total of 162 clones were isolated by plaque assay with Hz-AM1 cells from uncloned field populations of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) collected from Kenya, South Africa, Zimbabwe and Thailand. Restriction endonuclease (REN) analysis showed that 25 out of the 162 clones exhibited unique REN patterns, which should be characterized as variants of HearNPV. Five clones, NNg1, NS2, NMa1, NZ3 and NT1, were randomly selected from the 25 clones and characterized in both Hz-AM1 cells and H. armigera larvae, together with the clone G4 which was isolated in China and sequenced previously. In Hz-AM1 cells, clones NNg1, NMa1, NZ3 and NS2 produced more budded virions than clones G4 and NT1, whereas a higher amount of polyhedrin was produced in clones NNg1 and G4. The dose-mortality analysis revealed that clone NNg1 expressed the highest insecticidal activity against third instar H. armigera larvae, showing a 50% lethal dose (LD50) value of 10 occlusion bodies (OBs)/larva and a 50% lethal time (LT50) value of 4.0 days following peroral infection with OBs from infected larvae. Clones NS2, NMa1 and NZ3 had LD50 values of 32, 35 and 146 OBs/larva and LT50 values of 5.1, 4.4 and 4.8 days, respectively, indicating that these three clones had insecticidal activity somewhat lower than that of clone NNg1. In contrast, clones NT1 and G4 had markedly lower insecticidal activities compared with the four other clones, recording LD50 values of 826 and 3115 OBs/larva and LT50 values of 5.8 and 8.3 days, respectively. These results indicate that clone NNg1 is the most promising candidate for biological control programs of H. armigera larvae among the six clones characterized in this study.<br>

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