柞蚕核型多角体病毒对柞蚕蛹卵巢细胞感染性研究  

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

作　　者：王林美[1] 岳冬梅[1] 李树英[1] 范琦[1] 叶博[1] 赵振军[1] 张波[1] 

机构地区：[1]辽宁省农业科学院大连生物技术研究所,大连116024

出　　处：《生物技术通报》2013年第6期172-176,共5页Biotechnology Bulletin

基　　金：辽宁省自然科学基金项目(201102098);现代农业产业技术体系建设专项资金项目(ARS-22-02A)

摘　　要：研究柞蚕核型多角体病毒(ApNPV)与宿主细胞相互作用及形态发生机制,以便更好地开发利用杆状病毒表达系统。体外培养柞蚕蛹卵巢原代细胞,ApNPV病毒悬液感染细胞,测定细胞感染率及半数组织培养感染剂量,应用倒置相差显微镜和电子显微镜观察ApNPV感染后细胞的形态学变化。结果显示,ApNPV感染柞蚕蛹卵巢细胞5 d,倒置相差显微镜下观察即可发现细胞胀大,细胞内有多角体病毒出现;感染7 d,细胞内多角体病毒数量增多,细胞感染率为40.2%,细胞上清液中无包涵体病毒(NOV)效价滴度为6.95×105TCID50/mL。电子显微镜下观察到受病毒感染的细胞早期表现为细胞核膨大,而后核内出现病毒发生基质和核衣壳,核衣壳获得套膜形成病毒束,发育为成熟的病毒粒子,众多病毒粒子同时封存在一个折光性强,具有蛋白质性质、直径在0.7-10μm的多角体中。得出结论,ApNPV对柞蚕蛹卵巢细胞高度敏感,其在柞蚕蛹卵巢细胞内复制为典型的杆状病毒非同步复制。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.

关 键 词：柞蚕 核型多角体病毒 卵巢细胞 病毒感染 

分 类 号：S885.1[农业科学—特种经济动物饲养]