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

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％。

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.