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中国农学通报 ›› 2016, Vol. 32 ›› Issue (9): 125-130.doi: 10.11924/j.issn.1000-6850.casb15040097

所属专题: 生物技术

• 生物技术科学 • 上一篇    下一篇

In planta法在亚麻转基因中的应用

李杨,姜恭好,张倩,段海燕   

  1. 黑龙江大学生命科学学院,黑龙江大学生命科学学院,黑龙江大学生命科学学院,黑龙江大学农业资源与环境学院
  • 收稿日期:2015-04-13 修回日期:2016-02-29 接受日期:2015-12-23 出版日期:2016-03-29 发布日期:2016-03-29
  • 通讯作者: 段海燕
  • 基金资助:
    黑龙江大学博士启动金“分子标记技术在亚麻育种中的应用”;黑龙江省教育厅科学技术研究项目“高产型根叶两用白菜种质创新及分子标记辅助选择”(11551375);黑龙江大学高层次人才(创新团队)支持计划“东北水稻耐盐碱研究”(Hdtd2010-18)。

Application of in planta in Flax Transgene

李杨,, and 段海燕   

  • Received:2015-04-13 Revised:2016-02-29 Accepted:2015-12-23 Online:2016-03-29 Published:2016-03-29

摘要: 为了建立简单快速、高效稳定的亚麻遗传转化体系,本实验用亚麻的种子胚作为外植体,采用原位转化(in planta)法对亚麻转基因的实验条件(预培养时间、共培养时间、As浓度、silwett浓度和抽真空时间)进行了系统优化,结果确定最佳组合为:预培养时间8h,共培养时间72h,AS浓度100μmol/L,silwett浓度0.1%,抽真空时间5min。摸索适宜的草丁膦最低致死剂量用于亚麻转化植株的抗药性筛选,并对存活的植株进行PCR鉴定,成功获得转基因阳性植株。本研究实现了in planta技术在亚麻种子胚的转基因体系优化,有较大的实际应用价值。

关键词: 地下水, 地下水, 重金属, 环境风险, 大宝山矿区

Abstract: In order to establish a simple, quick, efficient and stable system for flax (Linum ustitatissimum L.) transformation, flax seed embryos were used as explants, the experiment conditions (pre-incubation time, coculture time, As concentration, silwett concentration and vacuum time) were optimized with in planta. The best combination was determined as: pre-incubation time of 8 h, co-culture time of 72 h, As concentration of 100 μmol/L, silwett concentration of 0.1%, vacuum time of 5min. The minimum lethal dose of glufosinate was found out for herbicide screening on transformed flax plants. PCR identification on surviving plants showed that positive transgenic plants were acquired successfully. The optimization of in planta transformation into flax seed embryos was achieved, and had great value in practical application.

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