凤仙花坏死斑病毒的RT-PCR和巢式PCR检测  

Detection of Impatiens necrotic spot tospovirus(INSV) by RT-PCR and Nested PCR

作　　者：张巧萍[1,2] 丁元明[1] 王云月[2] 李 周剑[1] 段禄华[1] 白永华[1] 孙兵绍[1] 

机构地区：[1]云南出入境检验检疫局,昆明650228 [2]云南农业大学植物保护学院,昆明650201

出　　处：《华中农业大学学报》2009年第1期23-26,共4页Journal of Huazhong Agricultural University

基　　金：国家“十一五”科技支撑计划项目(2007BAD45B04)资助

摘　　要：根据凤仙花坏死斑病毒(Impatiens necrotic spot tospovirus,INSV)S RNA上的核衣壳蛋白(N)基因序列(登录号为AB109100)保守区设计了3对特异性引物。应用常规RT-PCR和巢式PCR方法从表现同心圆症状的文心兰植株上检测得到了预期DNA片段,且巢式PCR检测灵敏度比常规PCR高。序列分析结果表明,该病毒的N基因序列和已经发表的凤仙花坏死斑病毒(登录号为AB109100、AB207803、DQ425096)核苷酸序列同源性为99%,确定表现同心圆症状的文心兰上携带了凤仙花坏死斑病毒。The specific primers were designed based on the conservative region of the N gene of INSV（AB109100）. The virus causing chlorotic ringspots on oncidium was identified as Impatiens necrotic spot tospovirus （INSV） through both RT-PCR and the nested PCR amplification technology, which generated a product of the correct predicted size. The sensitivity of the nested PCR is better than normal RT-PCR. The sequencing results showed the homology of N gene sequences between the INSV and the known species is 99%, which confirmed pathogen of this disease is INSV. This paper provide RT-PCR and the nested PCR for INSV detection,which may serve for plant inspection and quarantine.

关 键 词：凤仙花坏死斑病毒 RT-PCR 巢式PCR 

分 类 号：S41-33[农业科学—植物保护] S681.1