植物学报 ›› 2014, Vol. 49 ›› Issue (2): 183-189.DOI: 10.3724/SP.J.1259.2014.00183

• 技术方法 • 上一篇    下一篇

RNAi载体导入黄瓜的遗传转化体系

王烨, 顾兴芳*, 张圣平, 苗晗, 陈国华, 谢丙炎   

  1. 中国农业科学院蔬菜花卉研究所, 北京 100081
  • 收稿日期:2013-01-26 修回日期:2013-04-12 出版日期:2014-03-01 发布日期:2014-01-07
  • 通讯作者: 顾兴芳
  • 基金资助:

    国家现代农业产业技术体系专项资金资助项目;农业部园艺作物生物学与种质创新重点实验室资助项目

Transformation of RNAi Vector in Cucumber (Cucumis sativus) In Vitro by Agrobacterium tumefaciens-mediated Transfection

Ye Wang, Xingfang Gu*, Shengping Zhang, Han Miao, Guohua Chen, Bingyan Xie   

  1. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2013-01-26 Revised:2013-04-12 Online:2014-03-01 Published:2014-01-07
  • Contact: Xingfang Gu

摘要: 以黄瓜(Cucumis sativus)子叶和子叶节为外植体, 利用农杆菌(Agrobacterium tumefaciens)介导法建立黄瓜遗传转化体系并进行优化, 将南方根结线虫(Meloidogyme incognita)寄生相关基因的RNA干扰载体导入黄瓜, 通过潮霉素(Hyg)浓度梯度筛选得到99株潮霉素抗性植株。经PCR和Southern-blot检测, 获得10株目的基因以单拷贝形式整合的转基因黄瓜, 子叶和子叶节的Southern-blot阳性率分别为13.9%和6.9%。该研究建立并优化了RNA干扰载体导入黄瓜的高效遗传转化体系, 成功获得了转基因黄瓜植株。

关键词: 黄瓜, RNAi载体, 子叶, 子叶节, 转基因

Abstract: The RNAi vector of the target gene was transfected into cucumber (Cucumis sativus) cotyledons and cotyledon nodes by Agrobacterium tumefaciens-mediated transfection. A total of 99 regeneration plants were hygromycin-resistant. PCR and Southern-blot analysis revealed that target gene was integrated into the genome of 10 regenerated plants. The transformation efficiency in the cucumber cotyledon and cotyledon node was significantly increased from 4.1% to 13.9% and 3.1% to 6.9%, respectively, when using different hygromycin concentrations in the selection cultivation. We have optimized the genetic transformation system with an RNA interference vector in cucumber and successfully obtained transgenic cucumber plants.

Key words: cucumber, RNAi vector, cotyledon, cotyledon node, transformation