影响根癌农杆菌介导的木薯遗传转化因素分析  

Analysis on the Factors Influencing Transformation of Cassava (Manihot esculenta Crantz) Mediated by Agrobacterium tumefaciens

作　　者：姚庆荣[1] 郭运玲[2] 孔华[2] 贺立卡[2] 郭安平[2] 

机构地区：[1]甘肃行政学院,兰州730070 [2]中国热带农业科学院热带生物技术研究所,海口571101

出　　处：《植物研究》2012年第2期227-231,共5页Bulletin of Botanical Research

基　　金：中国热带农业科学院科技基金项目(RKY0724);中国热带农业科学院中央公益性科研院所基本科研业务费专项(ITBBZD2007-2-3)

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200μmol.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%。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 improv6d by using the following transformation conditions ： GV3101 as A. tumefaciens strains, infection time 30 -45 min, c0-culture time 3 - 4 d, engineering bacterium concentration （ OD^0 ） 0.45 and application of 200 jxmol ~ L- J AS. However, pre-cuhure showed negative influence on the transient expression rate. Eight transgenic cassava plants from 453 explants were obtained in this study PCR and Southern blot analysis. The transformation and the integration of glgC336 transgene was confirmed by frequency was 1.77%.

关 键 词：木薯 农杆菌介导 遗传转化 

分 类 号：S533[农业科学—作物学]