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Susceptibility of a Cloned Cell Line from Helicoverpa armigera to Homologous Nucleopolyhedrovirus

Sudawan Chaeychomsri 1, Win Chaeychomsri 2,3, Motoko Ikeda 2,3, and Michihiro Kobayashi 2,3
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
3. Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan
Abstract—A cloned cell line designated B-5 was isolated from the parental HAPO2 cell line, which derived from pupal ovarian tissues of Helicoverpa armigera. This cell line was cultured in Grace’s medium supplemented with 10% fetal bovine serum (FBS), and continuously subcultured at 4-day intervals. The population doubling time of this cloned cell line during the logarithmic phase was 22.8 h. RAPD and DAF analyses confirmed the identity of the B-5 cell line as H. armigera and clearly distinguished it as a new, unique cell line derived from the pupal ovaries. The susceptibility of B-5 cells to the H. armigera nucleopolyhedrovirus (HearNPV) was high and a higher yields of occlusion bodies (OBs) were obtained when compared to the parental HAPO2 cells. Infection of HearNPV in B-5 cells was successfully completed and produced the infectious OBs. Ultrastructural studies of infected H. armigera tissues revealed evidence of virus infection and OBs formation in the infected fat cells. The results and observations strongly indicated that B-5 cells were fully permissive for HearNPV infection. The data described in this study indicate that B-5 cell line is a highly productive new cloned cell line and will be useful for the production of HearNPV, and other applications in biotechnology.
 
Index TermsHelicoverpa armigera, nucleopolyhedrovirus, virus replication, insect cell culture, single-cell cloning, cell line characterization

Cite: Sudawan Chaeychomsri, Win Chaeychomsri, Motoko Ikeda, and Michihiro Kobayashi, "Susceptibility of a Cloned Cell Line from Helicoverpa armigera to Homologous Nucleopolyhedrovirus," Journal of Advanced Agricultural Technologies, Vol. 4, No. 3, pp. 200-208, September 2017. Doi: 10.18178/joaat.4.3.200-208
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