摘要
以新会橙、锦橙(CitrusSinensisOsb.)胚状体以及北碚447锦橙、纽荷尔脐橙(C.SinensisOsb.)田间成年树腋芽为转化起始材料,经含外源抗菌肽基因(ShivaA和CecropinB)的根癌农杆菌感染后,比较了胚状体在不同培养基中丛芽的诱导频率和卡那霉素(Km)质量浓度对胚状体丛芽生长以及最终转化频率的影响,也对成年树腋芽在不同BA、IAA质量浓度处理、不同创伤部位和是否用石蜡带(Parafilm)包裹创伤口的诱芽和转化率进行了研究。PCR检测和Southernblot分析证明外源抗菌肽基因(ShivaA和CecropinB)已导入上述4个柑桔品种;离体抑菌试验证明转基因植株叶片提取液对溃疡病菌有一定的抑制作用。建立了一个简便、快捷、通用的柑桔遗传转化体系。
The embryoids of Jincheng orange and Xinhui orange (Citrus sinensis Osbeck) were induced from nucellar embryo, and the mature shoots were obtained from 20-year-old plants of Beibei447 orange and Navel orange. After they were inoculated with agrobacterium strain LBA4404 harboring vectors that contained Shiva A and Cecropin B gene, the effects of various media, concentrations of kanamycin, places of shearing and parafilm on the shoot induction rate and transformation frequency were studied. PCR (polymerase chain reaction) and Southern blot analysis verified that the Shiva A and Cecropin B gene were transferred into the genome of citrus. Transgenic plants were grafted onto 6-month-old Trifoliate orange in greenhouse.
出处
《热带作物学报》
CSCD
2004年第1期11-16,80,共7页
Chinese Journal of Tropical Crops
基金
"九五"农业部重点科研计划"高新技术与基础研究"项目(编号:95农-15-03-02)
重庆市科技攻关项目资助。
关键词
根癌农杆菌
菌株介导
柑桔
遗传转化技术
citrus embryoid mature shoot agrobacterium-mediated transformation
