| 任 芳, 张尊平, 范旭东, 胡国君, 李正男, 董雅凤*.葡萄病毒B CP基因植物表达载体构建及烟草遗传转化[J].植物保护,2018,44(3):11-16. |
| 葡萄病毒B CP基因植物表达载体构建及烟草遗传转化 |
| Construction of a plant expression vector of the Grapevine virus B coat protein gene and transformation into Nicotiana tabacum |
| 投稿时间：2017-06-22 修订日期：2017-10-13 |
| DOI：10.16688/j.zwbh.2017236 |
| 中文关键词: 葡萄病毒B 外壳蛋白基因 植物表达载体 烟草 遗传转化 |
| 英文关键词:Grapevine virus B coat protein gene plant expression vector tobacco genetic transformation |
| 基金项目:国家现代农业产业技术体系建设专项资金(CARS-29-bc-1) |
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| 中文摘要: |
| 葡萄病毒B Grapevine virus B (GVB)是葡萄皱木复合病中栓皮病的病原, 为开展抗GVB转基因研究, 本研究利用RT-PCR技术克隆GVB外壳蛋白(coat protein, CP)基因, 与植物表达载体pRI 101-AN连接构建植物表达载体pRI-GVB CP。采用电击转化法将植物表达载体pRI-GVB CP导入农杆菌LBA4404, 并利用农杆菌介导的叶盘转化法将外源基因导入西方烟‘37B’。共获得16个烟草再生株系, PCR检测其中3个株系为阳性, 阳性株系播种获得的64株T1代植株中有30株扩增到目的条带, 阳性率为46.9%, 表明目的基因GVB cp 成功导入烟草并可成功遗传到子代。28株T1代转基因植株接种病毒进行抗病性鉴定, 其中有6株对接种病毒GVB具有抗性。 |
| 英文摘要: |
| Grapevine virus B (GVB) is the pathogen of grapevine corky bark disease in the rugose wood complex. In order to develop transgenic plants resistant to GVB, the coat protein (CP) gene of GVB was cloned and inserted into the plant expression vector pRI 101-AN and the recombinant plant expression vector pRI-GVB CP was constructed. The vector pRI-GVB CP was transformed into Agrobacterium tumefaciens LBA4404 by electric shock process, and then introduced into Nicotiana occidentalis ‘37B’ by Agrobacterium -mediated leaf disc transformation. Totally 16 regenerated tobacco strains were obtained in this study. PCR detection indicated that three stains were positive, and 46.9 percent of 64 T1 plants were positive by PCR detection, indicating that the target gene GVB cp was successfully introduced into the tobacco and successfully inherited to the progeny. The resistance of transgenic tobacco was identified by inoculating GVB into T1 transgenic plants. The results indicated that 6 out of 28 T 1 transgenic plants showed resistance to GVB. |
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