摘要
采用根癌农杆菌介导的叶盘转化法，将ＣＭＶＢＳ（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）１００μｇ／ｍＬ浓度下转化体叶片分化形成愈伤组织；用ＰＣＲ和ＤＡＳＥＬＩＳＡ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ＣＭＶＢＳ、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。
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 μg/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 challenging 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 the severity of systemic symptoms, and the degrees of protection were positive correlated with the extents of the coat protein expressed. It was further shown that when the transgenic plants were challenged with five concentrations (8,20,50,100 or 200 times of dilution of diseased leaf sap extract with buffer solutions) of CMV BS,CMV 27 and CMV 37 respectively, the protection was decreased with the increasing of concentration of the virus inoculum.
出处
《中国病毒学》
CAS
CSCD
1998年第3期257-262,共6页
Virologica Sinica
关键词
黄瓜花叶病毒
外壳蛋白基因
转基因烟草
抗病性
CMV banana strain, Coat protein gene, Transgenic tobacco plant, Strain, Protection
