摘要
为有效地控制贵州兰花的病毒，将经ELISA方法检测为齿兰环斑病毒（ORSV）阳性的蝴蝶兰样品摩擦接种至其枯斑寄主苋色藜进行纯化，RT-PCR 克隆并连接至 pMD18-T 载体后测序，用 DNA-MAN7．0和MEGA5．0软件进行核苷酸序列分析。结果表明：成功克隆该病毒CP基因，其序列长度为477 bp，编码158个氨基酸残基；序列同源性分析显示，该分离物的CP基因与已报道的11种ORSV各地分离物序列同源性高达96％～99％，而与同属其他3种病毒CP基因同源性均低于70％。证明，贵州蝴蝶兰上感染的病毒为 ORSV。
In order to provide a reference for effectively controlling Guizhou orchid virus,a suspected odontoglossum ringspot virus （ORSV）was detected by ELISA and isolated from P.amabilis in Guizhou, and purified by Chenopodium amaranticolor.The coat protein （CP）gene of the isolate was then cloned with RT-PCR,sequenced by pMD18-T vector and analyzed by DNAMAN7.0 and MEGA 5.0 softwares. The results showed that the sequence length of CP gene was 477 bp,and encoded a 158-amino acid CP protein.Result of sequence homogenous analysis showed that the isolate shared 9 6%~9 9% homology with 11 reported ORSV isolates,while less than 70% with other three viruses within the same genus （tobamovirus）.Therefore,it could be proposed that the virus isolated from P.amabilis was ORSV.
出处
《贵州农业科学》
CAS
北大核心
2014年第8期96-98,共3页
Guizhou Agricultural Sciences
基金
长江学者和创新团队发展计划"喀斯特山地生物多样性保护与可持续利用"(PCSIRT1227)
贵州省科技计划项目"贵州省植物生理与发育调控重点实验室建设"[黔科合计Z字(2011)4005]
贵阳市星火计划项目"贵阳高档花卉病毒检测技术及脱毒苗生产优化"[(2010)筑科农合同字第1-农-23号]
贵州省社发攻关项目"植物病毒诊断试剂盒的研制"[黔科合SZ字(2011)3100]
贵州省农业攻关项目"高档花卉病毒侵染及防治技术研究"[黔科合NY字(2010)3031]
贵州省科技联合基金项目"高档花卉杜鹃
蝴蝶兰和玫瑰病毒快速检测技术研究"[黔科合J字LKS(2010)17]
关键词
齿兰环斑病毒
外壳蛋白
分子鉴定
蝴蝶兰
贵州
odontoglossum ringspot virus
coat protein
molecular identification
Phalaenopsis amabilis
Guizhou
