| 烟草花叶病毒丁香分离物的分离与鉴定 |
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| 引用本文： | 黄金光,邓丛良,范在丰,田国忠,李怀方.烟草花叶病毒丁香分离物的分离与鉴定[J].植物病理学报,2004,34(3):215-220. |
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| 作者姓名： | 黄金光 邓丛良 范在丰 田国忠 李怀方 |
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| 作者单位： | 1 中国农业大学植物病理系, 北京 100094;2 中国林业科学院森林生态环境保护研究所, 北京 100091 |
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| 基金项目： | 科技部基础性工作项目，引进国际先进农业科技计划(948计划) |
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| 摘 要： | 从表现花叶症状的丁香病株上获得一病毒分离物,其在电镜下为约300 nm×18nm的杆状粒子;电泳分析表明感病组织中ds RNA大约为6.4kbp,而其外壳蛋白分子量约为17.6k Da。以上实验结果初步将该病毒分离物鉴定为烟草花叶病毒属(Tobamovirus)。根据该属病毒复制酶基因序列设计通用引物,进行RT-PCR检测,扩增出约1000 bp的预期特异片段(Gen Bank AY566703)。将PCR产物克隆后测序,序列分析表明,与从蚕豆中分离的TMV-B株系序列(Gen Bank AJ011933.1)同源性为99.90%。根据烟草花叶病毒(Tobacco mosaic virus,TMV)的RNA CP基因序列设计引物,进行RT-PCR,扩增出约800 bp的预期特异片段(Gen Bank AY56672),序列分析表明,与TMV-B株系序列(Gen Bank AJ011933.1)同源性达99%,上述实验结果表明,该病毒分离物为TMV。由于该分离物与TMV-B在指示植物上的症状存在明显差异,所以,作者把该分离物暂命名为TMV-S。
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| 关 键 词： | 丁香 烟草花叶病毒 鉴定 序列分析 |
| 文章编号： | 0412-0914(2004)03-0215-06 |
| 修稿时间： | 2004年1月6日 |
Isolation and identification of Tobacco mosaic virus infecting Syringa oblate |
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| HUANG Jin-guang,DENG Cong-liang,FAN Zai-feng,TIAN Guo-zhong,LI Huai-fang.Isolation and identification of Tobacco mosaic virus infecting Syringa oblate[J].Acta Phytopathologica Sinica,2004,34(3):215-220. |
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| Authors: | HUANG Jin-guang DENG Cong-liang FAN Zai-feng TIAN Guo-zhong LI Huai-fang |
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| Institution: | 1 Department of Plant Pathology, China Agricultural University, Beijing 100094, China;2 Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China |
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| Abstract: | Rod-shaped virus particles about 300 nm×18 nm were isolated from Syringa oblate expressing systemic mosaic in leaves. The double-stranded RNA pattern revealed one single band of about 6.4 kbp. The virus contained coat protein of approximately 17.6 kDa. According to these results, the virus isolate was identified as a member of Tobamovirus. A pair of primers were designed based on Tobamovirus RNA RdRp gene, and a fragment of about 1 000 bp(GenBank AY566703)was amplified by RT-PCR. The PCR product was cloned and sequenced. The sequence shared 99.90% homology with TMV-B strain (GenBank Accession No. AJ011933.1). Another pair of primers was designed based on TMV RNA CP gene. A fragment of about 800 bp(GenBank AY566702)was amplified. The recombinant plasmid was obtained and sequenced. The sequence shared 99% homology with TMV-B strain (GenBank Accession No. AJ011933.1). CP gene sequence and amino acids shared 99.37%, 100% homology with TMV-B strain respectively. On the basis of above results, the virus isolated from S. oblate in Beijing is Tobacco mosaic virus (TMV). |
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| Keywords: | Syringa oblate Tobacco mosaic virus (TMV) identification sequencing |
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