荧光定量PCR分析不同家蚕品种对质型多角体病毒(BmCPV)的感染抵抗性  

Analysis on Resistance of Various Bombyx mori Varieties to Cytoplasmic Polyhedrosis Virus Infection by Fluorescent Quantitative PCR

作　　者：吴萍[1,2] 钱荷英[1] 徐安英[1] 陈涛[1,2] 侯启瑞[1,2] 郭锡杰[1] 李龙[1,2] 

机构地区：[1]江苏科技大学蚕业研究所,江苏镇江212018 [2]农业部蚕桑产业产品质量监督检验测试中心,江苏镇江212018

出　　处：《蚕业科学》2013年第2期289-294,共6页ACTA SERICOLOGICA SINICA

基　　金：现代农业产业技术体系专项(No.CARS-22);江苏省自然科学基金项目(No.BK2010353);镇江市科技支撑计划项目(No.NY2011028);江苏科技大学引进人才科研启动项目(No.35181001)

摘　　要：根据家蚕质型多角体病毒(BmCPV)的多角体蛋白基因保守序列设计特异性引物,建立家蚕质型多角体病毒的实时荧光定量PCR检测方法,并应用于检测分析该病毒在蚕体内的增殖规律和筛选抵抗性强的家蚕品种资源。运用该方法检测分析BmCPV感染不同家蚕品种4龄起蚕后的动态增殖变化:大多数品种的幼虫感染BmCPV后24 h,中肠组织中的病毒已开始复制,感染后48～96 h病毒的增殖急速上升,至96 h达到高峰,感染后120 h病毒多角体蛋白基因的表达量显著下降。基于实时荧光定量PCR的检测结果分析显示家蚕品种间对BmCPV的抵抗性差异很大,10个供试品种中,热带多化性品种7005及热带二化性品种P50的抵抗性最强,欧系一化性品种4008和4017的抵抗性最弱。According to the conserved sequence of polyhedrin gene in silkworm(Bombyx mori) cytoplasmic polyhedrosis virus(BmCPV),a pair of specific primers were designed to establish a real-time fluorescent quantitative PCR method for detection of BmCPV and were applied in detection and analysis of BmCPV proliferation in silkworm and screening of silkworm variety resources with strong resistance.By this method,dynamic multiplication of BmCPV was analyzed after infection in the newly exuviated larvae of 4th instar from different silkworm varieties.The results showed that,in most tested silkworm varieties,the replication of BmCPV started at 24 h after infection in midgut tissue,rapidly increased at 48 ～96 h and reached the peak at 96 h.The expression level of BmCPV polyhedrin gene was significantly decreased at 120 h.The real-time fluorescent quantitative PCR results suggest that the resistibility to BmCPV varies greatly with silkworm varieties.Among the 10 tested silkworm varieties,the resistibility of 7005(tropic,polyvoltine) and P50(tropic,bivoltine) to BmCPV is the highest and that of 4008 and 4017(European,monovoltine) is the lowest.

关 键 词：家蚕质型多角体病毒 感染增殖 荧光定量PCR 家蚕品种 抵抗能力 

分 类 号：S884.51[农业科学—特种经济动物饲养]