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| 葡萄卷叶伴随3型病毒和葡萄A病毒的多重检测及其系统进化分析 | |
| 引用本文： | 王建辉,刘建军,陈克玲,李洪雯,席德慧,林宏辉.葡萄卷叶伴随3型病毒和葡萄A病毒的多重检测及其系统进化分析[J].园艺学报,2011,38(12):2401-2410. |
| 作者姓名： | 王建辉 刘建军 陈克玲 李洪雯 席德慧 林宏辉 |
| 作者单位： | 1. 生物资源和生态环境教育部重点实验室,四川大学生命科学学院,成都610064／四川省农业科学院园艺研究所,成都610066／农业部西南地区园艺作物生物学与种质创制重点实验室,成都610066 2. 四川省农业科学院,成都,610066 3. 四川省农业科学院园艺研究所,成都,610066 4. 生物资源和生态环境教育部重点实验室,四川大学生命科学学院,成都610064 |
| 基金项目： | 国家自然科学基金项目(31071669); 四川省财政基因工程专项(2011JYGC05-018) |
| 摘 要： | 葡萄卷叶伴随3型病毒（Grapevine leafroll associated virus-3，GLRaV-3）和葡萄A病毒（Grapevine virus A，GVA）常常复合侵染部分主栽欧美杂交葡萄品种。以半定量RT-PCR与qRT-PCR方法研究了‘希姆劳特’葡萄（Vitis vinifera L. × V. labrusca L.‘Himrod’）中不同组织内病毒的相对积累量，结果表明叶脉和韧皮部内的病毒相对积累量显著高于其他组织。以成熟枝条韧皮部组织的cDNA为模板，优化了RT-PCR多重扩增体系，可以同时扩增待测样本中两种病毒的目标基因。从一株复合感染GLRaV-3与GVA的‘藤稔’葡萄（Vitis vinifera L. × V. labrusca L.‘Fujiminori’）中，分别克隆了GLRaV-3外壳蛋白（coat protein，CP）基因全长序列和GVA部分基因组区域序列，后者包括CP基因的部分序列与病毒RNA沉默抑制子（viral suppressor of RNA silencing，VSR）基因全长序列。病毒核酸同源性分析与系统进化研究表明，不同GLRaV-3分离物的CP高度保守；而‘藤稔’葡萄的GVA分离物包含多种变异株，具有准种病毒的特点。 |
| 关 键 词： | 葡萄 葡萄卷叶伴随3型病毒 葡萄A病毒 病毒外壳蛋白基因 病毒RNA沉默抑制子基因 RT-PCR多重检测 病毒变异株 |
| 收稿时间： | 2011-8-9 |
| 修稿时间： | 2011-10-24 |
Optimizing Multiple Detection and Phylogenetic Studies on Grapevine leafroll associated virus-3 and Grapevine virus A |
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| WANG Jian-hui,,LIU Jian-jun,CHEN Ke-ling,LI Hong-wen,XI De-hui, LIN Hong-hui.Optimizing Multiple Detection and Phylogenetic Studies on Grapevine leafroll associated virus-3 and Grapevine virus A[J].Acta Horticulturae Sinica,2011,38(12):2401-2410. | |
| Authors: | WANG Jian-hui LIU Jian-jun CHEN Ke-ling LI Hong-wen XI De-hui LIN Hong-hui |
| Institution: | WANG Jian-hui1,2,3,LIU Jian-jun4,CHEN Ke-ling2,LI Hong-wen2,XI De-hui1,and LIN Hong-hui1(1Key Laboratory of Bio-resources and Eco-environment,Ministry of Education,College of Life Science,Sichuan University,Chengdu 610064,China,2Horticulture Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610066,3Key laboratory of Horticultural Crops Biology and Germplasm Enhancement in Southwest Regions,Ministry of Agriculture,4Sichuan Academy of Agricultural Sciences,Ch... |
| Abstract: | Co-infection of Grapevine leafroll associated virus-3（GLRaV-3）and Grapevine virus A（GVA）was a common phenomenon on some cultivars of Vitis vinifera L. × Vitis labrusca L. in Sichuan Province. Semi-quantitative RT-PCR and qRT-PCR were used to investigate the relative accumulations of the aforementioned two viruses in different tissues of infected grape trees（Vitis vinifera L. × V. labrusca L. ‘Himrod’）. The results suggested the relative accumulations of viruses in petiole and phloem were higher than that of others tissues. The multiple RT-PCR was developed to amplify the target genes of GLRaV-3 and GVA in phloem tissues of mature shoots（Vitis vinifera L. × V. labrusca L.‘Fujiminori’）simultaneously. The full length of coat protein gene（CP）of GLRaV-3 and partial genome region of GVA（including partial sequence of CP and full length of viral suppressor of RNA silencing gene，VSR）were cloned from V. vinifera L. × V. labrusca L.‘Fujiminori’separately. The nucleotide homology analysis and phylogenetic studies on different isolates of each virus were possibly revealed that CP from different GLRaV-3 isolates was highly conserved，whereas GVA Sichuan isolate contained very divergent variants and thus it could be quasispecies. |
| Keywords: | grape Grapevine leafroll associated virus-3 Grapevine virus A coat protein virus silencing suppressor multiple RT-PCR variant |
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