Spodoptera frugiperda multiple nucleopolyhedrovirus(SfMNPV),belonging to the species Alphabaculovirus spofrugiperdae,has been recently registered as an insecticide in China.This virus has a specific effect on the glob...Spodoptera frugiperda multiple nucleopolyhedrovirus(SfMNPV),belonging to the species Alphabaculovirus spofrugiperdae,has been recently registered as an insecticide in China.This virus has a specific effect on the global major agricultural pest Spodoptera frugiperda.To gain insights into viral infection,replication processes,and the complex formation of viral particles,in vitro studies using cell lines are essential tools.Although the IPLB-Sf9 and IPLB-Sf21 cell lines derived from S.frugiperda are widely used for studies on the infection and replication mechanisms of Autographa californica multiple nucleopolyhedrovirus(AcMNPV),their capacity to produce viral polyhedra after SfMNPV infection is not optimal.To address this limitation,a novel cell line named IOZCAS-Sf-1 was developed from a S.frugiperda population in Yunnan,China.The mitochondrial COX1 gene analysis confirmed the species origin of the IOZCAS-Sf-1 cell line.Furthermore,a comparative study was carried out to contrast the COX1 gene sequence of this novel cell line with that of IPLB-Sf9,highlighting the distinctions between the two.Importantly,the IOZCAS-Sf-1 cells exhibited a remarkable ability to generate polyhedra when infected with AcMNPV and SfMNPV,respectively.Consequently,this cellular lineage is considered a promising and valuable resource.It serves not only to investigate the molecular mechanisms of viral replication and its impact on host cells,but also to explore the transfection efficiency of SfMNPV DNA.This exploration further expands into its potential application in recombinant DNA experiments,laying a theoretical groundwork for the advancement of more effective biopesticides and sustainable agricultural practices.展开更多
为了研究Bm K IT提高Ac MNPV抗虫活性的作用机制,将Bm K IT基因插入到Ac MNPV中形成重组病毒。采用重组单价病毒Ac MNPV-Bm K IT(IE1)、Ac MNPV-Bm K IT(P10)、Ac MNPV-Bm K IT(PH)和一种重组双价病毒Ac MNPV-Bm K IT(P10)-vcath(PH),...为了研究Bm K IT提高Ac MNPV抗虫活性的作用机制,将Bm K IT基因插入到Ac MNPV中形成重组病毒。采用重组单价病毒Ac MNPV-Bm K IT(IE1)、Ac MNPV-Bm K IT(P10)、Ac MNPV-Bm K IT(PH)和一种重组双价病毒Ac MNPV-Bm K IT(P10)-vcath(PH),通过四唑盐比色法(MTT)和蛋白质印迹法(Western Blot)分析了Bm K IT在Ac MNPV的3个启动子调控下对Sf9细胞增殖和细胞凋亡的影响,结果显示,感染病毒36,48 h Bm K IT在不同启动子调控下表达量从高到低依次为PH、P10、IE1。同时分析了Ac MNPV介导的Bm K IT与组织蛋白酶的协同表达对昆虫Sf9细胞增殖和调亡的机制,结果表明,Ac MNPV-Bm K IT(P10)-vcath(PH)处理组对Sf9细胞抑制率比Ac MNPV-Bm K IT(P10)处理组平均提高了14.5%,凋亡相关蛋白c-Myc、cleaved-Caspase3、Bax表达量增加,Bcl-2表达量减少。展开更多
基金funded by the National Key Research and Development Program of China(Grant number 2022YFD1400700)Initiative Scientific Research Program,Institute of Zoology,Chinese Academy of Sciences(Grant number 2023IOZ010).
文摘Spodoptera frugiperda multiple nucleopolyhedrovirus(SfMNPV),belonging to the species Alphabaculovirus spofrugiperdae,has been recently registered as an insecticide in China.This virus has a specific effect on the global major agricultural pest Spodoptera frugiperda.To gain insights into viral infection,replication processes,and the complex formation of viral particles,in vitro studies using cell lines are essential tools.Although the IPLB-Sf9 and IPLB-Sf21 cell lines derived from S.frugiperda are widely used for studies on the infection and replication mechanisms of Autographa californica multiple nucleopolyhedrovirus(AcMNPV),their capacity to produce viral polyhedra after SfMNPV infection is not optimal.To address this limitation,a novel cell line named IOZCAS-Sf-1 was developed from a S.frugiperda population in Yunnan,China.The mitochondrial COX1 gene analysis confirmed the species origin of the IOZCAS-Sf-1 cell line.Furthermore,a comparative study was carried out to contrast the COX1 gene sequence of this novel cell line with that of IPLB-Sf9,highlighting the distinctions between the two.Importantly,the IOZCAS-Sf-1 cells exhibited a remarkable ability to generate polyhedra when infected with AcMNPV and SfMNPV,respectively.Consequently,this cellular lineage is considered a promising and valuable resource.It serves not only to investigate the molecular mechanisms of viral replication and its impact on host cells,but also to explore the transfection efficiency of SfMNPV DNA.This exploration further expands into its potential application in recombinant DNA experiments,laying a theoretical groundwork for the advancement of more effective biopesticides and sustainable agricultural practices.
基金supported by grants from the National Natural Science Foundation of China(No.31272100,31372199)the Natural Science Foundation of Shanxi Province,China(No.2014011038-1)+1 种基金the National High Technology Research and Development Program of China(863 Program,No.2012AA020809)the Program for the Top Young Academic Leaders of Higher Learning Institutions of Shanxi Province,China
文摘为了研究Bm K IT提高Ac MNPV抗虫活性的作用机制,将Bm K IT基因插入到Ac MNPV中形成重组病毒。采用重组单价病毒Ac MNPV-Bm K IT(IE1)、Ac MNPV-Bm K IT(P10)、Ac MNPV-Bm K IT(PH)和一种重组双价病毒Ac MNPV-Bm K IT(P10)-vcath(PH),通过四唑盐比色法(MTT)和蛋白质印迹法(Western Blot)分析了Bm K IT在Ac MNPV的3个启动子调控下对Sf9细胞增殖和细胞凋亡的影响,结果显示,感染病毒36,48 h Bm K IT在不同启动子调控下表达量从高到低依次为PH、P10、IE1。同时分析了Ac MNPV介导的Bm K IT与组织蛋白酶的协同表达对昆虫Sf9细胞增殖和调亡的机制,结果表明,Ac MNPV-Bm K IT(P10)-vcath(PH)处理组对Sf9细胞抑制率比Ac MNPV-Bm K IT(P10)处理组平均提高了14.5%,凋亡相关蛋白c-Myc、cleaved-Caspase3、Bax表达量增加,Bcl-2表达量减少。
