华北农学报 ›› 2008, Vol. 23 ›› Issue (6): 54-57. doi: 10.7668/hbnxb.2008.06.012

所属专题: 黄瓜 生物技术

• 论文 • 上一篇    下一篇

黄瓜花粉管通道法抗虫基因导入及卡那霉素抗性筛选

魏爱民, 张文珠, 杜胜利, 韩毅科, 张桂华, 刘楠   

  1. 天津科润黄瓜研究所, 天津, 300192
  • 收稿日期:2008-06-10 出版日期:2008-12-28
  • 作者简介:魏爱民(1972-),女,天津蓟县人,副研究员,主要从事黄瓜生物技术研究。
  • 基金资助:
    天津市自然科学基金项目(05YFJMJC07000)

Induction of Insect-resistant Gene into Cucumber by Pollen Tube Pathway and Selection of Kanamycin Resistance

WEI Ai-min, ZHANG Wen-zhu, DU Sheng-li, HAN Yi-ke, ZHANG Gui-hua, LIU Nan   

  1. Tianjin Kernel Cucumber Research Institute, Tianjin 300192, China
  • Received:2008-06-10 Published:2008-12-28

摘要: 研究了不同浓度卡那霉素(Km)对未转化黄瓜种子萌发及幼苗生长的影响,建立了转基因黄瓜卡那霉素抗性筛选体系。种子首先播在含Km 200 mg/L的1/2MS培养基中,能有效地抑制黄瓜幼苗生长,10 d后剪掉根系扦插移栽于蛭石中,能促使非转基因植株衰弱或死亡,有效地加速筛选。应用该筛选体系对花粉管通道法获得的抗虫转基因黄瓜T0种子进行初步筛选,获得Km抗性植株,抗性植株筛选率为1.6%,其中8.9%经PCR分子检测为阳性植株,初步证实EQKAM抗虫基因已整合到黄瓜基因组中。

关键词: 黄瓜, 花粉管通道法, 卡那霉素

Abstract: Effects of Kanamycin of different concentration on cucumber seed germination and seedling growth were studied. Screening method for kanamycin resistant seedling was established. Seeds were sowed first in 1/ 2MS medium containing Km 200 mg/ L,seedling growth could be effectively inhibited. After 10d,root was cut and then transplanted the seedling into Km free vermiculite could enhance seedling declining and dying,and therefore enhanced screening of resistant plants. T0 seeds from pollen tube pathway transformation were screened by this method,1. 6 % of seedlings were tested as resistant,among them 8.9 % were tested as PCR negative. It primarily certified that EQKAM gene was combined into Cucumber gene group.

Key words: Cucumber (Cucumis sativus L.), Pollen tube pathway, Kanamycin

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引用本文

魏爱民, 张文珠, 杜胜利, 韩毅科, 张桂华, 刘楠. 黄瓜花粉管通道法抗虫基因导入及卡那霉素抗性筛选[J]. 华北农学报, 2008, 23(6): 54-57. doi: 10.7668/hbnxb.2008.06.012.

WEI Ai-min, ZHANG Wen-zhu, DU Sheng-li, HAN Yi-ke, ZHANG Gui-hua, LIU Nan. Induction of Insect-resistant Gene into Cucumber by Pollen Tube Pathway and Selection of Kanamycin Resistance[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(6): 54-57. doi: 10.7668/hbnxb.2008.06.012.

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