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提高小麦基因枪法转化频率的研究

Studies of Improving the Frequency of Wheat Transformation by Biolistic Bombardment

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【作者】 梁辉; 唐顺学; 张弛; 赵铁汉; 李良材; 欧阳俊闻; 田文忠; 王道文; 贾旭;

【Author】 LING Hui; TANG Shun-Xue; ZHANG Chi; ZHAO Tie-Han; LI Liang-Cai;OUYANG Jun-Wen; TIAN Wen-Zhong; WANG Dao-Wen; JIA Xu(Institute of Genetics Chinese Academy of Sciences Beijing 100101)

【机构】 中国科学院遗传研究所!北京; 100101; 中国科学院遗传研究所!北?;

【摘要】 用基因枪法将带Bar-GUS双标记基因的质粒(pAHC25)转入普通春小麦品种中─60634的幼胚盾片,并获转基因植株。在轰击的经预培养3~4天的342块幼胚盾片再经筛选再生的植株中,经PCR和Southern分析表明,外源基因已稳定整合到小麦基因组中,转化率为0.88%。此外发现,幼胚培养基中用2mg/Ldicamba代替2mg/L2,4-D,可提高愈伤组织的再生能力。同时,为了保持小麦幼胚愈伤组织的分化能力,在诱导愈伤组织阶段5mg/Lbialaphos的筛选时间以不超过1个月为宜,及早转入3mg/Lbialaphos分化及壮苗培养基筛选转基因植株。

【Abstract】 In the present study immature embryos of spring wheat Zhong-60634 were bombarded with gold particles coated with pAHC25 containing Bar and GUS genes. A total of 17 bialaphos-tolerant plants were obtained from 342 immature embryos which were first cultured for 3-4 days and then bombarded and selected. Integration of the introduced genes into the genome of bosgenic wheat plants was shown by PCR and Southern analysis and transformation frequency was 0.88%. Furthermore, the present study showed that calli regeneration ability was improved while the embryo culture medium was supplemented with 2mg/L dicamba instead of 2mg/L 2,4-D. Meanwhile,in order to keep the calli differentiation ability calli should be transfered to differentiation meditum supplemented with 3mg / L bialaphos from calli selection medium with 5mg/L bialaphos within a month.

  • 【文献出处】 遗传学报 ,ACTA GENETICA SINICA , 编辑部邮箱 ,1999年06期
  • 【分类号】Q789
  • 【被引频次】47
  • 【下载频次】180
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