ToMV引起番茄系统性坏死与移动蛋白相关.docVIP

  ToMV 引起番茄系统性坏死与移动蛋白相 关# 张海峰，王婷，刘妮娜，程晓东，廖乾生** 5 10 15 20 25 30 35 40 45 （浙江理工大学生命科学学院生物工程研究所，杭州 310018） 摘要：番茄花叶病毒（Tomato mosaic virus，ToMV）是烟草花叶病毒属 (Tobamovirus)的典 型成员，是一类世界性分布的最具有经济重要性的植物病毒之一。ToMV-N5 株系能引起寄 主系统坏死，而 ToMV-S1 使寄主出现花叶症状。为了解 ToMV 引起寄主系统坏死的分子机 制，文中分别以两个株系病毒 RNA 为模板，利用 RT-PCR 扩增获得其全长 cDNA，进而通 过分段克隆的方式将其基因组克隆至携带有 35S 启动子的载体 pCB301 中，构建了农杆菌侵 染性克隆 pToMV-S1 和 pToMV-N5，验证了其具有侵染活性。并在此基础上构建了交换两 株系 MP 的嵌合病毒侵染性克隆 pToMV-N5MPS1 以及 pToMV-S1MPN5，pToMV-S1MPN5 与 ToMV-N5 的症状反应相一致，而 ToMV-N5 的 MP 被 S1 株系的 MP 取代后则表现为花叶 症状，这表明 ToMV-N5 引起番茄系统性坏死与其 MP 相关。Northern blot 分析病毒的基因 组含量表明，pToMV-S1 基因组含量低于 pToMV-N5，而其 MP 被 N5 株系的代替后其含量 大大提高，这表明 ToMV-N5 株系的移动蛋白能够提高病毒的含量，而且 ToMV-N5 株系引 起番茄系统性坏死可能由于其病毒含量高所引起。 关键词：植物病理学；番茄花叶病毒；番茄；系统性坏死；农杆菌侵染性克隆；移动蛋白 中图分类号：Q945.8 Systemic necrosis on Solanum lycopersicum infected by Tomato mosaic virus related to movement protein ZHANG Haifeng, WANG Ting, LIU Nina, CHENG Xiaodong, LIAO Qiansheng (Institute of Bioengineering, School of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018) Abstract: Tomato mosaic virus (ToMV) is the type species of the genus Tobamovirus and one of the most important economically plant viruses. ToMV-N5 showed systemic necrosis on the host plant while ToMV-S1 just showed wild mosaic. Complete genomic RNAs of both ToMV strains were obtained by RT-PCR respectively and were cloned into vector pCB301 which carrying duplicated 35S promoter. The agrobactium infectious clones of pToMV-S1 and pToMV-N5 were proved to be infectious. MP between that of ToMV-N5 and ToMV-S1 was exchanged by digestion and ligation to get the chimeric virus pToMV-N5MPS1 and pToMV-S1MPN5. The chimeric virus pToMV-S1MPN5 which carrying ToMV-N5 MP can cause systemic necrosis on host plants; while chimeric virus pToMV-N5MPS1 which carrying ToMV-S1 MP just induced mild mosaic symptoms expression on the seedlings of tomato. Thus, the systemic necrosis phenotype on the host plant might be related to the movement protein. Northern Blot assay showed tha