黄瓜花叶病毒外壳蛋白基因转化番茄的研究

Studies on Transformation of Lycopersicon esculentum with Cucumber Mosaic Virus Coat Protein Gene

通过根癌农杆菌介导 ,采用叶盘转化法 ,探讨了黄瓜花叶病毒外壳蛋白 (CMV CP)基因导入番茄栽培品种‘红宝石’的适宜条件 .结果表明 :适宜的卡那霉素 (Km)浓度、根癌农杆菌浓度、根癌农杆菌感染时间以及共培养时间分别为 35mg/L、D60 0 约 0 .5、10～ 15min和 2～ 3d ;将诱导形成的抗性愈伤组织转接到含有 35mg/LKm的分化培养基和生根培养基中使其分化不定芽和生根 ,获得了抗Km的番茄再生植株 .对再生植株进行再次离体培养抗Km鉴定和PCR分子检测 ,初步证明CMV CP基因已导入番茄再生植株的基因组中 .

By means of the leaf disk transformation with the help of Agrobacterium tumefaciens , several factors for transferring cucumber mosaic virus coat protein (CMV CP) gene into tomato cultivar `Hongbaoshi'were studied. The results showed that the appropriate concentration of kanamycin(Km), the concentration of A. tumefaciens, the soaking time in liquid medium containing A. tumefaciens and the co cultivation time were 35 mg/L, D 600 ≈0.5, 10～15 min and 2～3 d respectively. Calli from cotyledon explants grown on the medium containing 35 mg/L Km were transferred to the same medium for shoot regeneration, Km resistant plantlets were obtained on the root inducing medium. By testing the Km resistant ability of transformed plantlets cultured on the medium with 35 mg/L Km again and performing PCR assay, it was confirmed preliminarily that the CMV CP gene was integrated into the genome of tomato plantlet.