CSBV的分离鉴定及其感染幼虫后的应答反应OA北大核心CSTPCD
Isolation and Characterization of Chinese Sacbrood Bee Virusand Experimental Infection in Larvae
本研究旨在为了解江西某蜂场蜜蜂是否感染中蜂囊状幼虫病病毒(CSBV)并探究CSBV感染中蜂幼虫后的应答反应.采集该蜂场疑似感染CSBV的蜜蜂,采用RT-PCR的方法检测CSBV核酸,取阳性的蜜蜂样品进行病毒分离纯化、电镜观察及测序.将纯化鉴定后的CSBV对3日龄的中蜂幼虫进行人工感染,进行转录组测序,分析中蜂幼虫感染CSBV的应答反应.通过RT-PCR验证和电镜观察成功鉴定CSBV,并命名为CSBV-JX.CSBV-JX电镜观察直径为25～30 nm.对其进行分段扩增测序,发现基因组序列为8781 bp,对其进行系统发育分析,表明东方蜜蜂的SBV(AcSBV)与西方蜜蜂的SBV(AmSBV)明显分为两个分支,具有较强的地域分化性.转录组测序分析发现,中蜂幼虫在感染CSBV-JX后2 h有59个基因上调表达,83个基因下调表达;感染后12 h有68个基因上调表达,86个基因下调表达.本研究成功分离鉴定一株中蜂囊状幼虫病毒并命名为CSBV-JX.同时对CSBV-JX感染中蜂幼虫的转录组学分析发现,CSBV-JX感染2 h和12 h引起宿主的响应较小,这可能是CSBV-JX感染的2 h和12 h为病毒增殖的早期阶段病毒载量低,引起的宿主免疫响应小所导致.在CSBV-JX感染中蜂幼虫的2 h和12 h期间引起的差异基因与代谢通路关联较大,研究结果为阐明解析CSBV感染中蜂的分子机制提供了基础.
The objectives of the present study were to investigate whether the bees on an apiary were infected with Chinese Sacbrood bee virus(CSBV)and to explore the infection features in Apis cerana.The bees suspected to be infected with CSBV were collected from the apiary and the presence of CSBV nucleic acid was examined by RT-PCR.The positive bee samples were collected for virus isolation and purification,electron microscope observation and sequencing.Three-day-old Apis cerana were experimentally infected with the purified and identified CSBV and transcriptome sequencing was performed to analyze the infection features of Apis cerana.As a result,a CSBV-JX strain was isolated and identified through RT-PCR and electron microscope observation.The diameter of CSBV-JX was observed by an electron microscope to be 25-30 nm.The CSBV-JX was amplified and sequenced in segments and the genome sequence was found to be 8781 bp.Phylogenetic analysis showed that the SBV of the Apis cerana(AcSBV)and the SBV of the Apis mellifera(AmSBV)were divided into two branches with solid regional difference.Transcriptome sequencing analysis revealed that 59 genes were up-regulated and 83 were down-regulated in Apis cerana at 2 hours post infection with CSBV-JX and while 68 genes were up-regulated and 86 genes were down-regulated in Apis cerana at 12 hours post infection with CSBV-JX.In summary,this study,the CSBV-JX strain isolated and identified and minor responses to experimental infection in named CSBV-JX.Apis cerana were demonstrated at 2 hours and 12 hours by transcriptomics analysis.The reason for minor infection might be at the early stages of virus multiplication.In early infection stages,the viral loads were low,causing minimal proliferation and replication.However,the differentially expressed genes(DEGs)were highly related to the metabolic pathways in Apis cerana at 2 hours and 12 hours post infection.In conclusion,the results of this study provided a basis for illustrating the molecular mechanism of CSBV infection in Apis cerana.
费世港;刘合永;王叶元;冯敏;王子龙;孙京臣
华南农业大学动物科学学院,广州 510642江西农业大学蜜蜂研究所,南昌 330045
畜牧业
中蜂囊状幼虫病毒分离鉴定系统发育分析转录组学
Chinese Sacbrood virusisolation and identificationphylogenetic analysistranscriptomics
《中国动物传染病学报》 2024 (002)
89-96 / 8
广东省地方畜禽品种保护与利用促进项目(2018-143);国家自然科学基金项目(31860686)
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