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作 者 ：姚庆荣;郭运玲;孔华;贺立卡;郭安平*

期 刊 ：植物研究 2012年 32卷 2期 页码：227-231

关键词：木薯；农杆菌介导；

Keywords:cassava(Manihot esculenta Crantz), agrobacterium-mediated, transformation,

摘 要 ：采用根癌农杆菌介导的叶盘转化法，以我国南方地区主栽木薯品种—华南8号的胚状体子叶为受体，对影响木薯遗传转化效率的主要因素进行了分析。研究结果表明，在木薯的遗传转化中，选用GV3101作为浸染外植体的农杆菌菌株，将感染时间和共培养时间分别控制在30~45 min和3~4 d、菌液浓度（OD₆₀₀）采用0.45、并添加200 μmol·L^-1的乙酰丁香酮（AS）均可明显提高其转化效率,但若对外植体进行预培养反而会降低其转化效率。利用该体系从453块外植体中共转化获得10株抗性再生植株，经PCR和Southern杂交检测，有8株木薯的基因组中已整合进了外源基因glgC336，转化率为1.77%。

Abstract:Agrobacteriurn tumefaciens-mediated transformation system was used to study the factors influencing transformation in cassava. Histochemical GUS assay showed that the transient expression rate was significantly improved by using the following transformation conditions: GV3101 as A.tumefaciens strains, infection time 30-45 min, co-culture time 3-4 d, engineering bacterium concentration (OD₆₀₀) 0.45 and application of 200 μmol·L^-1 AS. However, pre-culture showed negative influence on the transient expression rate. Eight transgenic cassava plants from 453 explants were obtained in this study and the integration of glgC336 transgene was confirmed by PCR and Southern blot analysis. The transformation frequency was 1.77%.

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