Study on Proliferation Characteristics of Daphnis nerii cypovirus-23 in Sf9 Cells
KUANG Wen-Dong1, ZHAN Zhi-Gao1, GUAN Li-Mei1, WANG Jin-Chang1, YAN Cheng-Hua3, CHEN Jun-Hui1, LI Jiang-Huai1, ZHOU Xi2,*, JIN Liang1,*
1 Institute of Microbiology, Jiangxi Academy of Sciences, Nanchang 330096, China; 2 The State Key Laboratory of Virology, Wuhan 430071, China; 3 College of Life Sciences, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China
Abstract：A new type of Daphnis nerii cypovirus-23 (DnCPV-23) can kill a variety of Sphingidae insects and has the potential to be a new insect virus insecticide. At present, Daphnis nerii is the main model to study the virus, the replication of DnCPV-23 in cells is still unclear. In this study, DnCPV-23 virus particles were inoculated into Spodoptera frugiperda cell line Sf9, and the replication of the virus in the cells was analyzed. The virus-infected cells were cultured at 27 ℃, and the effect of the virus on the Sf9 cells were observed with the optical microscope; RNA of the infected cells on day 1, 3 and 7 was extracted and used as a template to obtain the cDNA, and the DnCPV-23 RNA level was analyzed by qRT-PCR; the supernatant of infected Sf9 cells and the serial passage of virus in Sf9 cells were studied by the qRT-PCR method. DnCPV-23 could adapt to Sf9 cells well, could successfully infect Sf9 cells, and produce cytopathic effect (CPE); virus genes could be detected in the cell culture supernatant; the virus could be passaged in Sf9 cells. DnCPV-23 could replicate and passage well on Sf9 cells and make the cells undergo CPE. The above-mentioned research provides a good cell model for in-depth study of DnCPV-23 invasion, transcription and other mechanisms and evaluation of virus virulence. At the same time, it provides a certain reference and basis for the development of new Sphingidae virus insecticides.
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