Biotechnology Bulletin ›› 2013, Vol. 0 ›› Issue (6): 172-176.

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Study on Infection of Pupal Ovaries Cells of Antheraea pernyi with ApNPV

Wang Linmei, Yue Dongmei, Li Shuying, Fan Qi, Ye Bo, Zhao Zhenjun, Zhang Bo   

  1. (Dalian Institute of Biotechnology, Liaoning Academy of Agricultural Sciences, Dalian 116024)
  • Received:2012-12-18 Revised:2013-06-20 Online:2013-06-20 Published:2013-06-20

Abstract: Mechamisms of interaction and morphogenesis of Antheraea pernyi somatic cells infected by Antheraea pernyi nuclear polyhedrosis virus(ApNPV)were studied for the further development of baculovirus expression system. Pupal ovaries cells from Antheraea pernyi were cultured in vitro followed by infection with Antheraea pernyi nuclear polyhedrosis virus.  Cells infecting rate and median tissue culture infective dose were determined. Cellular morphology was observed by an inverted phase contrast microscope and a light microscope. Results showed that, pupal ovaries cells were found expanding and nuclear polyhedrosis virus occurred in cells under the inverted phase contrast microscope after 5-days infection. After 7-days infection, the quality of nuclear polyhedrosis virus was found increased, cells infecting rate reached 40.2% and the titer of budded virus in cell supernatant was 6.95×105 TCID50/mL. An early sign of virus infection was enlargement of nuclei, then virogenic stroma and nucleocapsids were observed within the nucleus, nucleocapsids were enveloped and formed to virus bundls from the region between virogenic stroma and nuclear membrane, the well-developed virus were then enveloped into a strongly polarible proteinaceous polyhedra with a diameter of 0.7-10 μm under the electron microscope. It proved that ApNPV was highly sensitive to pupal ovaries cells and replicated asynchronously to form typical baculoviruses in the cells.

Key words: Antheraea pernyi, Nuclear polyhedrosis virus, Ovaries cells, Virus infection