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Characterization of the Helicoverpa Armigera Nucleopolyhedrovirus During Serial Passage in Cell Culture

Sudawan Chaeychomsri 1, Win Chaeychomsri 2, Motoko Ikeda 2, and Michihiro Kobayashi2
1. Central Laboratory and Greenhouse Complex, Faculty of Agriculture at Kamphaeng Saen, Kasetsart University, Kamphaeng Saen, Nakhon Pathom 73140, Thailand
2. Department of Zoology, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand
Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan
Abstract—Helicoverpa armigera nucleopolyhedrovirus (HearNPV) is highly pathogenic to the cotton bollworm, H. armigera. There have been several attempts to develop in vitro production of this virus, including the selection of high producing cell line. In order to evaluate the potential of in vitro methods of producing this virus on a large-scale, the virus was propagated in the H. armigera (HAPO2) and H. zea (HZ-AM1) cell lines and the correlation between the few polyhedra (FP) phenotype of HearNPV and the mutation in the polyhedrin (polh) and fp25k genes was analyzed. The many polyhedra (MP) phenotype was the predominant population of HearNPV for 3 passages in HZ-AM1 culture. The production of occlusion bodies (OBs) decreased with increasing passage number. The phenotypic observations on the HearNPV FP variants when this virus is serially passaged in HZ-AM1 cells suggest genomic changes occurring in the viral genes. A single point mutation resulting in an amino acid change was identified in the fp25k gene. This mutation could be correlated with the appearance of the FP phenotype with no relation to the polh gene. In contrast, serial passage in HAPO2 cells showed a slower accumulation of FP variants when compared to the use of HZ-AM1 cell line and no mutation was found in the fp25k gene. Therefore, HAPO2 cell line is a productive new cell line that will be useful for the scale-up process of HearNPV bioinsecticides.

Index Terms—Helicoverpa armigera, Helicoverpa zea, nucleopolyhedrovirus, insect cell culture, few polyhedra, fp25k gene.

Cite: Sudawan Chaeychomsri, "Characterization of the Helicoverpa Armigera Nucleopolyhedrovirus During Serial Passage in Cell Culture," Journal of Advanced Agricultural Technologies, Vol. 2, No. 1, pp. 63-70, June 2015. Doi: 10.12720/joaat.2.1.63-70
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