盆栽文心兰上的凤仙花坏死斑病毒的检测与分子鉴定

Detection and molecular identification of Impatiens necrotic spot virus isolated from potted Oncidium

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SRNA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HindIII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。

The virus causing chlorotie ringspots on ingoing potted oneidium 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.