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发根农杆菌对苦豆子高频转化条件的优化

Optimization of the Factors Affecting Genetic Transformation of Sopho-ra alopecuroides Mediated by Agrobacterium rhizogenes

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【作者】 赵东利张春广王新宇郑国锠

【Author】 ZHAO Dong-li1,2, ZHANG Chun-guang1 ,WANG Xin-yu1* , ZHENG Guo-chang1 (1. Institute of Cell Biology, Lanzhou University, Lanzhou 730000; 2. Medical College, Dalian University , Dalian 116622)

【机构】 兰州大学细胞生物学研究所兰州大学细胞生物学研究所 兰州 730000大连大学医学院 大连 116622兰州 730000兰州 730000

【摘要】 为优化发根农杆菌对药用植物苦豆子(Sophora alopecroides)遗传转化的条件,系统地研究了苦豆子外植体的种类(子叶、下胚轴、幼茎、幼叶),发根农杆菌的悬浮介质和密度、乙酰丁香酮的有元,以及共感染时的辅助处理方法(振动培养、真空渗透)等因素对转化率的影响。实验结果发现,(1)以子叶为外植体;(2)用MS培养基重悬菌体至光密度 0D600=0.6,并添加终浓度为 100μmol/L的乙酰丁香酮作为诱导菌液;和(3)真空(7.0 X104Pa)渗透 15 min,3者的组合可有效地提高发根农杆菌 A4对苦豆子的转化频率,最高频率可达到89.1%。

【Abstract】 In order to realize the high frequency transformation of Sophora alopecuroides mediated by Agrobacterium rhizogenes , this work dealt with the effects of the types of S. alopecuroides explant( cytoledon, hypocotyls, young stem and leaf segments), suspending medium and density of Agrobacterium rhizogenes A4 and some assistant methods (shaking, vacuum infiltration) during tissue-bacterium co-cultivation on transformation efficiency. As a result, the combination of the following three factors was found to be optimum in raising transformation frequency of A . rhizogenes to S. alopecuroides: a) using cotyledon as an explanf. b) A. rhizogenes was suspended with MS liquid medium supplemented with 100 umol/L acetosyringons (AS) to OD600 =0.6 and c) vacuum infiltration 15 min (7.0 x 104Pa) , by which the transformation frequency could be increased to 82.6% maximally.

  • 【文献出处】 药物生物技术 ,Pharmaceutical Biotechnology , 编辑部邮箱 ,2002年04期
  • 【分类号】Q943
  • 【被引频次】44
  • 【下载频次】221
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