SeMNPV抑制HaSNPV诱导的Se-UCR细胞凋亡

Helicoverpa armigera Nucleopolyhedrovirus Triggered Se-UCR Apoptosis is Blocked by Spodoptera exigua Nucleopolyhedrovirus

在采用共感染和共转染的方法构建扩大杀虫范围的重组病毒的研究过程中发现棉铃虫单核衣壳核多角体病毒(Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus,HaSNPV)能诱导甜菜夜蛾细胞Se-UCR发生典型凋亡,但不能诱导另一株甜菜夜蛾细胞Se-301产生凋亡。以5 MOI的HaSNPV感染Se-UCR,在12h左右可以观测到少量细胞凋亡,24h能观察到明显的凋亡,凋亡细胞数量随时间不断增加,到72h基本上所有的细胞均发生凋亡,成为凋亡小体,基因组DNA片段化。同时发现HaSNPV诱导的甜菜夜蛾Se-UCR细胞凋亡能够被甜菜夜蛾多核衣壳核多角体病毒(Spodoptera exigua multicapsid nucleoplyhedrovirus,SeMNPV)所抑制,进一步点杂交试验发现SeMNPV和HaSNPV共同感染Se-UCR获得了HaSNPV在该细胞中的复制。

Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV) and Spodoptera exigua mullicapsid nucleopolyhedrovirus (SeMNPV) are commercially produced as pesticides. As pesticides, both viruses have very narrow host range and can not make cross infection. Trying to generate a recombinant baculovirus with broader host range by co-infection and co-transfection, we found that HaSNPV could trigger apoptosis of a S. exigua cell line Se-UCR, which could be blocked by SeMNPV, but apoptosis did not occur in Se-301, a cell line also from S. exigua. When infected Se-UCR with 5 MOI, the apoptotic body was first found at 12 hpi, obvious apoptosis could be observed at 24 hpi, at the end almost all cells were dead at 72 hpi. Those observations were confirmed by genome DNA fragmen-tation analysis. Dot blot analysis indicated that SeMNPV could also help HaSNPV genome DNA and then virus replication in Se-UCR cells, but not in Se-301.