Productive replication of Malacosoma neustria nucleopolyhedrovirus (ManeNPV) in Md203 cell line

Authors : İsmail DEMİR, Nurten GÜREL, Remziye NALÇACIOĞLU, İkbal A İNCE, Zihni DEMİRBAĞ

Pages : 239-248

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Publication Date : 2009-12-01

Article Type : Research Paper

Abstract :A plaque-purified genotypic variant, derived from a field isolate of the Malacosoma neustria nucleopolyhedrovirus insert ignore into journalissuearticles values(ManeNPV); from Turkey was characterized based on in vitro replication properties in a cell line, Md203 derived from Malacosoma disstria. The life cycle of ManeNPV was studied based on the cytopathic effects insert ignore into journalissuearticles values(CPEs);, polyhedral inclusion body insert ignore into journalissuearticles values(PIB); formation, budded virus insert ignore into journalissuearticles values(BV); production, viral DNA replication and polyhedrin protein expression in ManeNPV-infected Md203 cells. Infection of Md203 cells with ManeNPV resulted in cytopathic effects within 24 and 36 h post infection and most cells detachted from the bottom of the culture plate on the fourth day. Typical NPV cytopathic effects insert ignore into journalissuearticles values(CPE); like granulated and rounded cells, nuclear hypertrophy and impairment in cell proliferation, and lost cell shape with the extendings were observed. Budded viruses were detected at 24 h p.i. However, significant increase in BV production was observed after 48 h p.i. PIB was firstly detected at 36 h p.i. When BV’s were used for reinfection of the Md203 cells, the result showed that these cells support production of viable virus progeny. We also observed a significant level of viral DNA synthesis by 24 h p.i. The results indicate that ManeNPV can be propagated at in vitro system for further studies and thus has a potential for development into an effective biotechnological and microbial insecticidal agent.
Keywords : Key words Baculovirus, Malacosoma neustria NPV, virus replication, Md203 cell line