CHEN Yuan-Sheng, XIAO Huo-Gen, ZHANG Shu-Guang and FAN Fu-Zhong. Genetic transformation of flue-cured tobacco cultivars G28 and K326 protecting to cucumber mosaic virus[J]. Virologica Sinica, 1998, 13(3).
Citation: CHEN Yuan-Sheng, XIAO Huo-Gen, ZHANG Shu-Guang, FAN Fu-Zhong. Genetic transformation of flue-cured tobacco cultivars G28 and K326 protecting to cucumber mosaic virus .VIROLOGICA SINICA, 1998, 13(3) : 262.

烤烟栽培品种G28和K326抗黄瓜花叶病毒的遗传转化

  • 采用根癌农杆菌介导的叶盘转化法,将CMVBS(香蕉株系)的CP基因转入广东烤烟栽培品种G28和K326,获得了再生转基因烟株。在卡那霉素(Km)100g/mL浓度下转化体叶片分化形成愈伤组织;用PCR和DASELISA分析,结果证明CP基因已整合到转化烟株的染色体中,并得到不同程度的表达。利用广东地区香蕉、烟草和番茄上的CMVBS、27和37株系,采用机械接种和蚜虫传毒法分别进行攻毒试验,结果证明转基因烟株对这三个株系的攻击都表现出明显的抗病能力,其抗性程度与烟株中的CP表达量呈正相关。采用五种攻毒浓度处理,发现随着攻毒浓度的增加,转基因烟株的抗病能力逐渐减弱。

Genetic transformation of flue-cured tobacco cultivars G28 and K326 protecting to cucumber mosaic virus

  • Two flue cured tobacco cultivars, G28 and K326, from Guangdong,were transformed by CMV BS CP gene from diseased banana via Agrobacterium tumefaciens , and 313 transgenic tobacco plants were obtained. Calli were observed on the small square sections of leaves of the transgenic plants in culture medium containing 100 g/mL Km,indicating the Km resistant marker genes (NPTⅡ) has been successfully introduced into the plants. It was confirmed with PCR analysis and DAS ELISA assay that the CMV BS CP genes were stably integrated into the genomes of the transgenic tobacco plants, and were subsequently found to be expressed in these plants. The transgenic tobacco plants were inoculated mechanically or by aphids with the hallenging CMV strains BS,27 and 37 from diseased banana,tobacco and cucumber in Guangdong,respectively. The transgenic tobacco plants expressing coat protein of BS were demonstrated protecting effect to the infection of these CMV strains significantly, delaying the disease expression and reducing

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    Genetic transformation of flue-cured tobacco cultivars G28 and K326 protecting to cucumber mosaic virus

    • 1. Institute of Virology.Wuhan University,Wuhan 430072

    Abstract: Two flue cured tobacco cultivars, G28 and K326, from Guangdong,were transformed by CMV BS CP gene from diseased banana via Agrobacterium tumefaciens , and 313 transgenic tobacco plants were obtained. Calli were observed on the small square sections of leaves of the transgenic plants in culture medium containing 100 g/mL Km,indicating the Km resistant marker genes (NPTⅡ) has been successfully introduced into the plants. It was confirmed with PCR analysis and DAS ELISA assay that the CMV BS CP genes were stably integrated into the genomes of the transgenic tobacco plants, and were subsequently found to be expressed in these plants. The transgenic tobacco plants were inoculated mechanically or by aphids with the hallenging CMV strains BS,27 and 37 from diseased banana,tobacco and cucumber in Guangdong,respectively. The transgenic tobacco plants expressing coat protein of BS were demonstrated protecting effect to the infection of these CMV strains significantly, delaying the disease expression and reducing

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