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作物学报 ›› 2011, Vol. 37 ›› Issue (03): 484-488.doi: 10.3724/SP.J.1006.2011.00484

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

应用RNA沉默技术获取抗黄瓜花叶病毒(CMV)和烟草花叶病毒(TMV)转基因烟草

牛颜冰,王德富,姚敏,闫钊,由文鑫   

  1. 山西农业大学生命科学学院，太谷 030801
  • 收稿日期:2010-07-30 修回日期:2010-11-26 出版日期:2011-03-12 网络出版日期:2011-01-17
  • 基金资助:

    本研究由国家自然科学基金项目(30571216)和山西省自然科学基金项目(20051071)资助。

Transgenic Tobacco Plants Resistant to Two Viruses via RNA Silencing

NIU Yan-Bing,WANG De-Fu,YAO Min,YAN Zhao,YOU Wen-Xin   

  1. College of Life Science, Shanxi Agricultural University, Taigu 030801, China
  • Received:2010-07-30 Revised:2010-11-26 Published:2011-03-12 Published online:2011-01-17

PDF

1648

被引次数

14

摘要： RNA沉默是迄今最为有效的抗病毒策略，利用该策略不但能获得免疫转基因植株，且所得植株不易与其他病毒基因重组或异源包壳、生物安全性较高。将本实验室已构建的携带有烟草花叶病毒(TMV)部分移动蛋白基因(ΔMP)和黄瓜花叶病毒(CMV)部分复制酶基因(ΔRep)反向重复结构的植物表达载体pBIN438-MP-Rep(i/r)，用农杆菌浸润法转化普通烟草品种K326，共得196株转化植株，经卡那霉素筛选和PCR检测发现128株为阳性转基因株；PCR-Southern和RT-PCR分析表明外源基因已整合到烟草基因组并在转录水平上得到表达；ELISA结果显示20.3%的转基因植株对CMV和TMV复合侵染表现免疫性。本结果为利用RNA沉默技术进行植物抗多种病毒育种提供重要数据，为防治其他多种病毒复合侵染提供借鉴。

关键词: RNA沉默, 黄瓜花叶病毒, 烟草花叶病毒, 双病毒抗性

Abstract: RNA silencing is a phenomenon of homologous RNA degradation induced by dsRNA, which is an effective strategy to obtain virus resistant plants so far. By means of this strategy, higher bio-security immune transgenic plants were obtained, avoiding recombining and transcapsidating with other virus genes. In this study, the recombinant plant expression vector pBIN438-MP- Rep(i/r)consisting the inverted repeat of TMV-ΔMP and CMV-ΔRep fusion fragment was transformed into tobacco cultivar K326 via Agrobacterium-mediated. The transformants were selected in the culture medium with 100 mg L–1 Kan. One hundred and ninety-six transgenic plants were obtained, One hundred and twenty-eight of which were positive plants, and the resistance to CMV and TMV was tested at the degree of virus. PCR-Southern blot and RT-PCR analysis of the transgenic plants demonstrated that the exogenous DNA was integrated into the tobacco genomic DNA and was expressed in transcriptional level. Resistance assay indicated that about 20.3% transgenic plants were immune to the co- infection with TMV and CMV. This result will provide to crops an important reference for plant anti-viral breeding and for preventing the viral co-infection.

Key words: RNA silencing, Cucumber mosaic virus, Tobacco mosaic virus, Dual-virus resistance

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