保定地区一种新辣椒病毒病的鉴定  

Identification of a new virus on sweet pepper in Baoding district of Hebei Province

作　　者：夏惠娟[1] 李志勇[2] 郭京泽[1] 李兴红[3] 

机构地区：[1]天津出入境检验检疫局,天津塘沽300456 [2]河北农业大学植物保护学院,河北保定071001 [3]北京农科院植保环保所,北京100089

出　　处：《河北农业大学学报》2006年第6期65-67,86,共4页Journal of Hebei Agricultural University

基　　金：北京市自然科学基金项目(6052010)

摘　　要：从保定东郊温室大棚中典型花叶症状的甜椒病株上得到1个病毒分离物（BD），经枯斑二三生烟分离纯化3次后，并保存在茄门甜椒上。通过酶联免疫方法、鉴别寄主反应、电镜观察测定等，确定保定分离物是辣椒轻斑驳病毒（Pepper mild mottle virus，PMMoV）。用Trizol提取病叶总RNA，反转录为cDNA，通过2对特异性引物，分别扩增出病毒外壳蛋白基因片段和与病毒复制相关蛋白基因片段。保定分离物的病毒外壳蛋白基因，与5个PMMoV分离物核甘酸同源性为94．5％～100％，而与3个TMV分离物核苷酸同源性为65．4％～67．7％，从而进一步验证了保定分离物为PMMOV。A few samples were collected from a green house in east suburb of Baoding district for virus identification. By 3 times of successive isolation from single necrotic lesion in Nicotiana tabacum cv. Samsun- NN, a virus isolate （short as BD） was obtained. By means of DAS- ELISA, electron microscopy observation and response of indicator hosts, the BD isolate was identified as Pepper mild mottle virus （PMMoV）. Then the total RNA of BD isolate was extracted and the eDNA was obtained by RT- PCR amplification. Using two pairs of specific primers, the coat protein gene and replication associated protein gene were amplified. Comparison of the nucleotide sequence of CP gene of BD isolate with five PMMoV isolates and three TMV isolates showed BD isolate was PMMoV.

关 键 词：辣椒轻斑驳病毒 双抗体夹心酶联免疫吸附反应 反转录PCR 辣椒 

分 类 号：S432.4[农业科学—植物病理学]