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盆栽文心兰上的凤仙花坏死斑病毒的检测与分子鉴定
引用本文:张巧萍,丁元明,李旻,周键,白永华,寸东义,曹云华.盆栽文心兰上的凤仙花坏死斑病毒的检测与分子鉴定[J].植物检疫,2008,22(6).
作者姓名:张巧萍  丁元明  李旻  周键  白永华  寸东义  曹云华
作者单位:1. 云南出入境检验检疫局,昆明,650228;云南农业大学
2. 云南出入境检验检疫局,昆明,650228
摘    要:应用酶联免疫吸附检测法(ELISA)和RT-PCR技术从表现有同心圆褪绿斑的进境盆栽文心兰上检测出凤仙花坏死斑病毒.ELISA检测发现该病毒在文心兰上分布不均匀,病毒主要集中在表现症状的病斑处.同时,根据该病毒S RNA上的核衣壳蛋白(N)基因序列保守区设计了一对特异性引物,利用RT-PCR方法检测得到500bp的预期DNA片段.经HindⅢI和EcolI酶切验证后克隆了该序列片段,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(登陆号为AB109100、AB207803、DQ425096)核苷酸序列同源性为99%,进一步确认进境盆栽文心兰上携带凤仙花坏死病毒.这是INSV首次在中国的报道,也是第一次在文心兰上发现该病毒.

关 键 词:文心兰  凤仙花坏死斑病毒(INSV)

Detection and molecular identification of Impatiens necrotic spot virus isolated from potted Oncidium
Zhang Qiaoping,Ding Yuanming,Li Min,Zhou Jian,Bai Yonghua,Cun Dongyi,Cao Yunhua.Detection and molecular identification of Impatiens necrotic spot virus isolated from potted Oncidium[J].Plant Quarantine,2008,22(6).
Authors:Zhang Qiaoping  Ding Yuanming  Li Min  Zhou Jian  Bai Yonghua  Cun Dongyi  Cao Yunhua
Abstract:The virus causing chlorotic ringspots on ingoing potted oncidium was identified as Impatiens necrotic spot virus(INSV)through both Enzyme-linked immunosorbent assay(ELISA)and RT-PCR amplification technology.The results of ELISA showed the distribution of INSV is enever on Oncidium,which mainly focus on lesion region.And then with the primers designed based on the conservative region of the INSV N gene,we can gain a 500bp fragment.We cloned the sequence by enzyme cutting both HindIII and EcolI,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 is first report INSV occur in china,this is also first repot INSV infected oncidium.
Keywords:RT-PCR
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