| 番茄不孕病毒BJ株系基因组测定与侵染性克隆 |
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| 引用本文： | 施伟,金圣塔,张海峰,王婷,陈集双,廖乾生.番茄不孕病毒BJ株系基因组测定与侵染性克隆[J].农业生物技术学报,2011,19(6). |
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| 作者姓名： | 施伟 金圣塔 张海峰 王婷 陈集双 廖乾生 |
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| 作者单位： | 浙江理工大学生命科学院 杭州310018 |
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| 基金项目： | 浙江省自然科学项目(No.Y3080228); 浙江理工大学科研启动项目资助 |
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| 摘 要： | 番茄不孕病毒(Tomato aspermy virus,TAV)可侵染包括藜科(Chenopodiaceae)、茄科(Solanaceae)等在内的24个双子叶家族和3个单子叶家族的100多种植物,是具有重要经济价值的植物病毒之一.为研究TAV BJ株系(Tomato aspermy virus,TAV-BJ)的基因组功能,本实验对TAV-BJ基因组克隆测序,并构建侵染性克隆.以TAV-BJ侵染心叶烟(ic otiana glutinosa)的总RNA为模板,RT-PCR获得其RNA2和RNA3；以TAV-BJ的dsRNA为模板,RT-PCR获得全长RNA1,目的片段PCR产物克隆测序获得TAV-BJ基因组全序列信息.RNA1全长3 409 nt,编码994个氨基酸的1a蛋白；RNA2全长3 023 nt,含2个开放阅读框(open reading frame,ORF),2a ORF编码829个氨基酸的2a蛋白,2b ORF编码78个氨基酸的2b蛋白；RNA3全长为2 216 nt,包含2个ORF,3a ORF编码247个氨基酸的3a蛋白,外壳蛋白(coat protein,CP)ORF 编码219个氨基酸的CP蛋白(TAV-BJ基因组RNA1、2和3 GenBank登录号分别为HQ424163,HQ424164和HQ424165).TAV-BJ基因组cDNA克隆体外转录成RNA并接种于心叶烟上,结果表明转录产物在寄主上的症状反应和TAV-BJ病毒粒子RNA的接种相一致,TAV-BJ基因组cDNA侵染性克隆具有活性.由TAV-BJ各个基因片段与缺失2b基因的黄瓜花叶病毒Fny株系(Cucumber mosaic virus,CMV-Fny△2b)构建的假重组病毒接种于心叶烟,结果显示TAV-BJ的RNA2和RNA3能恢复CMV-Fny△2b在寄主上症状反应.嵌合型RNA3F3aTcp和RNA3T3aFcp的症状反应结果表明,F1F2△2bRNA3 T3aFcp在寄主上产生花叶症状与F1 F2△2bT3相一致.本研究获得TAV-BJ的基因组序列,成功构建侵染性克隆,同时发现TAV-BJ的3a基因具有CMV-Fny的2b基因的某些功能.
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| 关 键 词： | 番茄不孕病毒 基因组序列 侵染性克隆 3a基因 |
Sequences and Infectious Clones of Tomato aspermy virus Strain Isolated from Beijing |
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| Shi Wei
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Jin Shengta
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Zhang Haifeng
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Wang Ting
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Chen Jishuang
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Liao Qiansheng.Sequences and Infectious Clones of Tomato aspermy virus Strain Isolated from Beijing[J].Journal of Agricultural Biotechnology,2011,19(6). |
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| Authors: | Shi Wei Jin Shengta Zhang Haifeng Wang Ting Chen Jishuang Liao Qiansheng |
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| Institution: | Shi Wei Jin Shengta Zhang Haifeng Wang Ting Chen Jishuang Liao Qiansheng* College of Life Sciences,Zhejiang Sci-Tech University,Hangzhou 310018,China |
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| Abstract: | Tomato mosaic virus(TAV) is an important pathogen in agriculture,infecting more than 100 species of 24 dicotyledoneae and 3 monocotyledoneae,such as Chenopodiaceae and Solanaceae.In order to analyze the role of genomic RNAs,full length sequence of TAV isolated from Beijing(TAV-BJ) was obtained by RT-PCR and infectious cDNA clones was constructed.Using total RNA from Nicotiana glutinosa infected by TAV-BJ as template,full length of RNA2 and RNA3 were amplified by RT-PCR,and RNA1 was obtained from cDNA of TAV... |
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| Keywords: | Tomato aspermy virus Sequence of genomic RNAs Infectious clones 3a gene |
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